Abnormal Karyotype Research Articles

MDS-255 Clinical Characteristics and Mutational Profile of Patients With Myelodysplastic Syndromes and Chronic Myelomonocytic Leukemia.

Next-generation sequencing (NGS) is a booming tool that allows the detection of useful characteristics for the diagnosis and prognosis of myeloid pathologies. An association between certain mutations and clinical characteristics has recently been proposed and predictive models of survival that contribute to the IPSS-R and cytogenetic risk have emerged. To describe clinical characteristics and their association with mutational profiles in patients with Myelodysplastic Syndromes (MDS) and Chronic Myelomonocytic Leukemia (CMML) diagnosis. Assess the usefulness of molecular scales to predict risk and survival. Retrospective cohort study, including adult patients diagnosed with MDS and LMMC from two highly complex private centers. 55 patients were included, 41% women, with a median age of 55 years (31-87). According to the WHO classification, 3 CMML; 27 MDS-DM; 6 MDS-DU; 6 MDS-EB-1; 2 MDS-EB-2, and 1 MDS-SA. Karyotype abnormalities were found in 16 patients (29%), being the alterations in chromosome 8 (8/55), 7 (7/55), and 5 (6/55) the most frequent. 71% (39/55) were classified as good prognosis in the reviewed cytogenetic risk group. According to the IPSS-R, they presented very low risk (20%), low (46%), intermediate (13%), high (7%), and very high (13%). 91% of the patients presented at least one somatic mutation by NGS. The median number of mutations was 3 (0-6). TET2, SRSF2, DNMT3A, ASXL1, and TP53 were the most frequently reported mutations, in 15, 13, 11, 10, and 8 patients respectively. Adverse Risk mutations were found in more than 50% (21/39) of patients with good cytogenetic risk. Of the 36 low/very low-risk patients according to IPSS-R 4/36 (11%) and 6/36 (17%) were categorized as high- and intermediate-risk, respectively, with the MIPSS-Scale. Patients with TET/IDH mutations had significantly higher cancer rates (p=0.040) and a higher trend of IE (p=0.062). The median OS for the entire group was 1.5 years, and at 2 years it was 27%. Both the Nazha stratification scale and the MIPSS-R were good predictors of OS (p=0.0021 and p=0.0008, respectively) in our population. The frequency and type of the mutations that we found were similar to those described in the literature. Mutations in TET/IDH are a special group of interest with potential clinical implications. The molecular risk scales of Nazha and MIPSS-R were useful tools to predict OS.

MDS-014 Case Report: Woman With Myelodysplastic Syndrome and Complex Karyotype, Unusual Presentation in Age, and Unfavorable Clinical Behavior

Myelodysplastic syndromes (MDS) are a group of heterogeneous pathologies that are characterized by clonal alterations in hematopoietic stem cells (HSC). MDS are usually seen in patients over 60 years of age and are accompanied by decreased ability of HSC to differentiate and enter apoptosis, producing bone marrow with increased cellularity conditioned by ineffective hematopoiesis and a degree of peripheral cytopenia. Approximately 30% of de novo MDS are associated with a complex karyotype, defined as the existence of 3 or more chromosomal abnormalities in the same karyotype. We present the case of a 27-year-old woman referred for pancytopenia, who has a son with aplastic anemia. The patient presented a 6-month evolution picture characterized by asthenia and adynamia. She presented with pale integuments and without clinical bleeding. Initial laboratories showed hemoglobin 4.9 g/dL, hematocrit 14%, mean corpuscular volume 96.6 fL, mean cell hemoglobin 33 pg, white blood cells 2,000 cells/mL, neutrophils 900 cells/mL, lymphocytes 900 cells/mL, monocytes 100 cells/mL, platelets 36,000, lactate dehydrogenase 357 U/L. Bone marrow aspirate was performed, reporting hypercellularity for age, heterogeneous cellularity, dyspoiesis in the red series, and blasts <5%. Flow cytometry reported 11% of total cells with CD2+, CD33+, CD34+, CD38+, CD45+, CD117+, and HLA+ phenotypic expression and changes in the maturation pattern of the granulocytic series. Karyotyping showed 348-73,XXX,sl,t(4:10)(q35;q11.2) x(2),add(12)(q24)x(2),1+2+3+4+5+6+7+8+9+10+11+12+13+14+15+16+17+18+19+20+21+22 (26)/46,XX(2)(28). The double cell line predominated over the one that contains polyploidies, involving translocation between the long arms of chromosomes 4 and 10 and adhesion in the long arms of chromosome 12 of indeterminate origin on the normal line. Myelodysplastic syndrome with excess blasts was diagnosed, and IPSS-2 high risk was concluded. During the patient's evolution, she presented a high transfusion requirement and recurrent infections, dying due to an infectious process. The age of the patient contrasts with reports in the literature that MDS tend to be more aggressive at a younger age. The complex karyotype in MDS predisposes patients to clonal evolution; despite this, it is not considered a separate prognostic factor by the IPSS-R. Ideally, this group of young patients should seek early hematopoietic stem cell transplantation with curative intent.

ALL-004 Philadelphia Chromosome-Positive T-Cell Acute Lymphoblastic Leukemia/Lymphoblastic Lymphoma: A Rare Case Report

T-acute lymphoblastic leukemia/lymphoblastic lymphoma (T-ALL/LBL) is a rare and aggressive leukemia. Philadelphia chromosome-positive (Ph+) T-ALL is exceedingly rare and has therapeutic and prognostic significance. The case study herein is a very rare presentation. A case report of a 52-year-old Egyptian female, who presented in 2012 with a 2-month history of fatigue, bone aches, and abnormal vaginal bleeding. Oncology Center Mansoura University. The patient appeared pale with significant splenomegaly. Complete blood count: white blood cell count 6.9 k/µL, microcytic hypochromic anemia with hemoglobin 10.4 g/dL, and platelet count 149 k/µL. A peripheral blood smear showed neutrophil toxic granulation. Bone marrow aspirate (BMA) was infiltrated with 45% blasts, and myeloid and erythroid series were suppressed. No significant dysplasia was noted. Flow cytometry analysis performed on BMA showed an abnormal population of blasts expressing TdT (20%), cytoplasmic CD3 (35%), CD7 (64%), CD2 (62%), CD5 (61%), CD1a (0.38%), CD4 (27%), CD8 (32%), CD10 (3.6%), CD117 (5.4%), CD 34 (2.41%), and CD79a (27%), revealing a case of ALL (T-cell lymphoblastic lymphoma/leukemia). The fluorescence in-situ hybridization (FISH) analysis performed on interphase nuclei obtained from a BMA sample revealed a dual-fusion hybridization signal pattern for BCR and ABL1 probes on chromosomes 9 and 22, indicating t(9:22) in 100%. Cytogenetic analysis revealed an abnormal complex karyotype, 46,XY,t(9;22),(q34;q11.2)[25], in all counted cells. BCR-ABL1 gene rearrangement by quantitative real-time PCR was 100% IS. The patient was diagnosed with a case of T-ALL/LBL with t(9;22);BCR-ABL1 (Ph+). The patient started the MRCUK protocol in combination with imatinib 400 mg/d. She finished maintenance therapy on 10/12/2016. The patient is still on imatinib 400 mg/d. The patient is still in complete remission and in deep molecular response with no detectable minimal residual disease. Ph+ T-ALL is rare. Differentiation between Ph+ T-ALL/LBL and T-LB crisis of chronic myeloid leukemia (CML) was difficult because it is diagnosed with no prior history of CML. The addition of tyrosine kinase inhibitors to intensive chemotherapy has greatly improved the outcomes of Ph+ ALL patients.

AML-139 Early Onset of Therapy-Related Acute Myeloid Leukemia in a Patient Treated for a Hodgkin Lymphoma

Therapy-related acute myeloid leukemia (t-AML) is a known complication of certain types of therapies, such as cytotoxic, radiation, or immunosuppressants. At present, t-AML incidence is relatively low. However, this is expected to change in the near and medium future due to improved overall survival rates. t-AML can occur a few months to several years after the initial therapy, depending on the type and dosage of therapy and patient-specific variation. The prognosis is generally poor, with patients presenting complex karyotype and significant structural abnormalities. As such, the optimal treatment remains a challenge. We report the case of a patient treated for a Hodgkin lymphoma stage IIB in December 2019, who underwent 8 cycles of ABVD and radiotherapy 30 Gy with a negative PET scan in August 2020, diagnosed with acute monoblastic leukemia (M5a - FAB) in January 2021. A 53-year-old male, smoker, presented with fatigue and oliguria for the past month. Laboratory studies revealed leukocytosis (272×109/L), thrombocytopenia (83×109/L) and no anemia, with 90% blasts in peripheral blood smear. The metabolic profile revealed a tumoral lysis syndrome and an elevated lactate dehydrogenase of 1877 U/L. The bone marrow aspirate showed 98-99% blasts cells of monocytic lineage while the flow cytometry analysis indicated 95% monocytic cells, positive for CD64+, CD33++, CD56+, CD36+ slab, CD300e-, CD14-, HLA-DR+. Normal karyotype, negative FISH for del 17p - TP53, and negative for the tested genes on RT-PCR (FLT3, RUNX1-RUNX1T1, CBFb-MYH11, MLL-AF9, PML-RARA), NGS negative (blood specimen). The patient had been successfully treated with conventional chemotherapy and underwent bone marrow transplantation from an unrelated donor with 100% donor chimerism at one year. t-AML only rarely presents with normal karyotype and there is a lack of clearly defined prognostic factors. Optimal treatment is still very much a matter of debate, in part because of the poor prognosis of the condition, and the only treatment with curative intent remains allogeneic hematopoietic stem cell transplantation.

MDS-472 STAG2 and SRSF2 Mutations Might Define Prognosis of MDS Patients With Isolated Trisomy 8

Isolated trisomy 8 is a karyotypic abnormality frequently found in hematological neoplasms. It is found in 10% of myelodysplastic syndromes (MDS) with cytogenetic aberrations. It is classified as an intermediate risk subgroup in MDS IPSS-R (Revised International Prognostic Scoring System) because of the heterogeneous clinical course of the patients. The aim of this study was to determine whether mutational profile could be useful to improve the risk reclassification of MDS with isolated trisomy 8. In a cohort of 591 MDS patients, we included 38 patients with isolated trisomy 8 confirmed through karyotype and FISH. We carried out next generation sequencing in bone marrow samples of these patients with an in-house custom panel with more than 100 myeloid-related genes. NGS files were processed with our own pipeline. Statistical analysis was performed with SPSS. A total of 102 somatic mutations were found in our cohort. The most frequently mutated genes were TET2, STAG2, ASXL1 and SRSF2. Interestingly, the mutational profile allows the differentiation of two subtypes of isolated trisomy 8: patients with mutations in either SRSF2 or STAG2 (n=16), and those without mutations in these genes (n=22). Both groups presented similar clinical characteristics. However, patients with STAG2 and/or SRSF2 mutations had a shorter time to progression to AML and lower overall survival than patients without these mutations (p<0.05). Surprisingly, the risk for AML progression and the overall survival of patients with STAG2/SRSF2 alterations was similar to those MDS classified as high risk in IPSS-R (p>0.05). Similarly, patients with trisomy 8 without these mutations did not show significant differences in survival and progression to AML when compared to the group of MDS with low IPSS-R risk (p>0.05). This study demonstrates, for the first time, that mutational profile of MDS with isolated trisomy 8 allows these patients to be reclassified into two groups based on the presence or absence of mutations in STAG2 and/or SRSF2. These alterations are clearly associated with an adverse prognosis. Therefore, we suggest the need to study the mutational profile of these patients in order to reclassify them on the prognostic scale.

Complete Androgen Insensitivity Syndrome

Androgen receptor gene mutations on Xq12, which also have a 46XY karyotype abnormality, are the root cause of the X-linked uncommon recessive disorder of sex development known as androgen insensitivity syndrome (AIS). Complete AIS existed as a female, with normal breast, no uterus, ovaries, and fallopian tube with a blind-ending vagina but the presence of bilateral undescended testis either in the inguinal canal, abdomen, or labioscrotal junction and elevated testosterone levels. This was a rare case of a 22-year-old female patient who presented with primary amenorrhea. Ultrasonography showed gonads in the mid parts of inguinal canals on both sides, reaching up to the superficial ring. On investigation, increased in level of serum testosterone, follicle-stimulating hormone along with the luteinizing hormone was seen. AIS is actually very disturbing to individuals and families, so close collaboration between radiologist, pathologist, treating consultants, and psychiatrists are required for the proper management.

Cytogenetic techniques in current biomedical research. PART III: numerical alterations of human karyotype

Numerical abnormalities of karyotype are the result of genome mutations. Unlike gene and chromosomal abnormalities, genome mutations do not disrupt the structure of DNA or chromosomes. The cause of numerical changes in the karyotype is a violation of the mechanism of chromosome segregation during meiosis or mitosis. Like other mutations, genome mutations are a natural mechanism for increasing of genetic diversity in offspring. At the same time, humans usually have negative effects of any numerical deviations from the norm, for this reason, cytogenetic examination of aneuploidies is an important diagnostic tool in medical genetics.A change in the number of sex chromosomes is usually not lethal. The spectrum of detected deviations in the carrier is from inconstant impairment of reproduction but a normal phenotype to malformations of some internal organs, infertility and severe intellectual disabilities. Aneuploidies of autosomes are always a threat to life and health. Only autosomal trisomies on chromosomes 13, 18, 21 and 22 are compatible with live birth, there are solitary reports of the birth of children with polyploidies. At the same time, the prognosis of life is relatively favorable only in the case of trisomy 21, leading to the formation of Down syndrome. Other aneuploidies usually lead to spontaneous termination of pregnancy in the early stages and are discovered in samples of abortion material.In this regard, cytogenetic analysis of chromosomal aneuploidies is used to establish the genetic cause of anomalies and malformations in the postnatal period, delays in speech and psychomotor development, reproduction disorders in adults. Of particular importance is the cytogenetic analysis of the karyotype of embryos in the prenatal period. The proposed lecture analyzes the mechanism of formation of genomic mutations and their diversity. The possible medical consequences of the presence of various types of aneuploidies are considered. To the reader attention is offered syndromes associated with a change in the number of chromosomes in the karyotype. The description is illustrated by real images of patient karyotypes.The lecture is aimed primarily at students of medical and biological specialties, young specialists who plan to use cytogenetic research methods in their practical activities, and doctors who are faced with the need to analyze and interpret the results of cytogenetic analysis. To assimilate the material under discussion, it is recommended to familiarize yourself with the previous lecture of the cycle.

Recurrent Pregnancy Loss and Associated Risk Factors

Background: As documented by the histopathology the recurrent pregnancy loss (RPL) is characterized as two or more miscarriages. The true prevalence of RPL is unknown, but is reported that two percent of women are affected by RPL per annum. A well-known risk factor for recurrent pregnancy losses is the high maternal age. Objective: This study was carried out to determine the risk factors and consequences of recurrent pregnancy loss. Study design: It is a prospective study Place and Duration: Conducted for the duration of one year from August 2021 to July 2022. The patients attended the obstetrician and gynecology department of Khawaja Muhammad Safdar Medical College, Allama Iqbal Memorial Teaching Hospital Sialkot. Material and Methods: This study was performed on 130 patients, admitted to tertiary care unit within the duration of one year. The written consent was taken from all the patients included in the study. The ethical and review board committee of the hospital approved the study. The women that had less than 2 miscarriages were excluded from the study. Results: The mean age of the mother at the time of admittance to hospital was 34 years and the mean of the miscarriages reported earlier in these patients were 2.9±1.3. The risk factors were found to be linked in most of the patients. The abnormality of the uterus and the thyroid dysfunction were some of the major risk factor in most of the patients with recurrent pregnancy loss. In case of thyroid dysfunction, the hyperthyroidism and hypothyroidism were studied and the results showed that the hyperthyroidism was present in case of 17% of the patients while hypothyroidism was found in 83% of the cases. Parental karyotype abnormality was also found in 4% of the cases. Conclusion: It was concluded that the patients suffering from recurrent pregnancy loss should underwent through the process of appropriate examination to clarify and conclude the risk factors involved in causing RPL. The protein S and factor XII deficiency may be playing a role in causing RPL, therefore LDA therapy can improve the rate of live birth in these patients. Keywords: recurrent pregnancy loss and protein S deficiency.

Prenatal genetic diagnosis of the fetuses with isolated corpus callosum abnormality

Objective: To explore the application value of chromosome karyotype analysis, chromosomal microarray analysis (CMA) and whole exome sequencing (WES) in prenatal diagnosis of isolated corpus callosum abnormality (CCA) fetus. Methods: Fetuses diagnosed with isolated CCA by ultrasound and MRI and receiving invasive prenatal diagnosis in Guangzhou Women and Children's Medical Center and Qingyuan People's Hospital from January 2010 to April 2021 were selected. Karyotype analysis and/or CMA [or copy number variation sequencing (CNV-seq)] were performed on all fetal samples, and WES was performed on fetal samples and their parents whose karyotype analysis and/or CMA (or CNV-seq) results were not abnormal. Results: Among 65 fetuses with isolated CCA, 38 cases underwent karyotype analysis, and 3 cases were detected with abnormal karyotypes, with a detection rate of 8% (3/38). A total of 49 fetuses with isolated CCA underwent CMA (or CNV-seq) detection, and 6 cases of pathogenic CNV were detected, the detection rate was 12% (6/49). Among them, the karyotype analysis results were abnormal, and the detection rate of further CMA detection was 1/1. The karyotype results were normal, and the detection rate of further CMA (or CNV-seq) detection was 14% (3/21). The detection rate of CMA as the first-line detection technique was 7% (2/27). A total of 25 fetuses with isolated CCA with negative results of karyotyping and/or CMA were tested by WES, and 9 cases (36%, 9/25) were detected with pathogenic genes. The gradient genetic diagnosis of chromosomal karyotyping, CMA and WES resulted in a definite genetic diagnosis of 26% (17/65) of isolated CCA fetuses. Conclusions: Prenatal genetic diagnosis of isolated CCA fetuses is of great clinical significance. The detection rate of CMA is higher than that of traditional karyotyping. CMA detection could be used as a first-line detection technique for fetuses with isolated CCA. WES could increase the pathogenicity detection rate of fetuses with isolated CCA when karyotype analysis and/or CMA test results are negative.

Abstract IA005: On the use of stem cells to generate sarcomas in vitro models

Abstract Sarcomas are rare cancers arising in soft tissue and bone. They are difficult to diagnose and treat, with 38-48% mortality for this often young age group. It is now generally accepted that molecular changes in cancer open avenues for personalized medicine. However, to achieve this for sarcoma there are several challenges, as representative model systems are scarce. Moreover, sarcomas are heterogeneous and &amp;gt;70 sarcoma types are recognized. A simple dichotomy based on molecular pathways of simple versus complex genomics is often used as a conceptual framework. My group has used mesenchymal stem cells to generate a model for complex genome sarcomas such as osteosarcoma. Murine and canine mesenchymal stem cells (MSCs) can be used to model spontaneous malignant transformation towards sarcomas with complex genomics. These MSCs have an abnormal karyotype, many structural variants and point mutations at whole genome sequencing analysis, and form sarcomas (osteosarcoma, undifferentiated pleomorphic and spindle cell sarcoma) after injection into mice. Cross-species analysis reveals that p53 loss is an early event in sarcomagenesis, and MSCs with a knockout in Trp53 transform earlier compared to wild-type MSCs. In addition, loss of p16Ink4a is shown to be a driver of osteosarcomagenesis: the majority of spontaneously transformed murine MSCs has alterations in p15Ink4b, p16Ink4a, or p19Arf and knockout of these genes in MSCs results in accelerated transformation. Of interest, osteosarcoma cells with defective p16INK4A are sensitive to the CDK4/CDK6 inhibitor Palbociclib, suggesting that a subset of osteosarcoma patients with intact Rb, but defective p16 or overexpression of CDK4 and/or CDK6 (in total 20-23%) might benefit from CDK4/CDK6 inhibition. Bone and soft tissue tumors with a simple genome often carry chromosomal translocations. We have generated a model for pseudomyogenic hemangioendothelioma (PHE), a rare locally aggressive vascular tumor. PHE has a tumor-specific t(7;19)(q22;q13) translocation resulting in a SERPINE1-FOSB fusion. We used CRISPR/Cas9 to generate this fusion in induced pluripotent stem cells (hiPSCs), which were subsequently differentiated into ECs (hiPSC-ECs) to address this. Comparison of parental with PHE hiPSC-ECs shows invasive growth in mice after transplantation, and transcriptome analysis identifies PI3K-Akt and MAPK signaling pathways as possible therapeutic targets. The modified hiPSC-ECs thus recapitulate functional features of PHE and demonstrate how these stem cell based translocation models can be used to understand tumorigenic mechanisms and identify therapeutic targets. Citation Format: Judith V.M.G. Boveé. On the use of stem cells to generate sarcomas in vitro models [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA005.

Characteristics and prognosis of children with recurrent T-cell acute lymphoblastic leukemia: a long-term follow-up report in China

This study assessed the relapse characteristics and prognosis of 145 children newly diagnosed with T-cell acute lymphoblastic leukemia (T-ALL). The overall complete response (CR) rate was 91.7% (133/145), and the overall recurrence rate was 31.6% (42/133). The recurrence rate in the intermediate-risk (IR) group and high-risk (HR) group was 15.4% and 47.1%, respectively (p < 0.001). Patients with young age, early T-cell precursor ALL, central nervous system (CNS) involvement, TCRγ gene rearrangement, karyotypic abnormalities, or absence of TCRβ gene rearrangement (p < 0.05) tended to relapse. All recurrences occurred within 36 months after diagnosis. The HR group recurred earlier than the IR group (p= 0.026). The 3-year overall survival (OS) rate was significantly lower in the HR group than in the IR group (p < 0.001). All relapsed children died within 12 months after recurrence. Early intervention may benefit children with a high risk of recurrence.

Reproductive outcomes following recurrent first-trimester miscarriage: a retrospective cohort study.

What are the subsequent reproductive outcomes (livebirths, miscarriages or other adverse pregnancy outcomes or no further pregnancy) of women with recurrent miscarriage (RM) attending a dedicated clinic? Of women with RM, 77% had a subsequent pregnancy, and among these pregnancies, the livebirth rate was 63%. RM affects ∼1-3% of women of reproductive age. RM has known associations with advanced maternal age, obesity, diabetes, inherited thrombophilias, thyroid dysfunction, endometriosis and parental balanced translocations. However, ∼ 50% of women or couples will be left without an explanation for their pregnancy loss, even after completing investigations. RM is also associated with secondary infertility and adverse pregnancy outcomes including preterm birth and perinatal death. We undertook a retrospective cohort study to identify subsequent pregnancy outcomes in women with RM, defined as three consecutive first-trimester miscarriages. Women attending the RM clinic at a tertiary university hospital in the Republic of Ireland over 12 years (2008-2020) with a confirmed diagnosis of primary or secondary first-trimester RM were eligible for inclusion. In total, 923 charts were identified for review against the eligibility criteria. Women with non-consecutive first-trimester miscarriages or ectopic pregnancy were excluded. Epidemiological and clinical information regarding medical history, investigation and management was gathered from paper and electronic medical records. Data were analysed using SPSS (Version 27). Associations between maternal characteristics and outcomes were explored using the χ2 test, with significance set at P < 0.05. Multinomial regression analysis was performed using a stepwise approach. There were 748 women who were included; 332 (44%) had primary RM and 416 (56%) had secondary RM. The median age was 36 years (range 19-47). Foetal aneuploidy was the most common investigative finding (15%; n = 111/748); 60% had unexplained RM. In addition to supportive care, most women were prescribed aspirin (96%) and folic acid (75%). Of the 748 women, 573 had a subsequent pregnancy (77%) and 359 (48% of all women; 63% of pregnancies) had a livebirth, while 208 had a further pregnancy loss (28% of all women; 36% of pregnancies) and 6 were still pregnant at the end of the study. Women aged 35-39 years were more likely to have a livebirth than no further pregnancy (relative risk ratio (RRR): 2.29 (95% CI: 1.51-5.30)). Women aged 30-34 years were more likely to have a livebirth (RRR: 3.74 (95% CI: 1.80-7.79)) or a miscarriage (RRR: 2.32 (95% CI: 1.07-4.96)) than no further pregnancy. Smokers were less likely to have a livebirth (RRR: 0.37 (95% CI: 0.20-0.69)) or a miscarriage (RRR: 0.45 (95% CI: 0.22-0.90)) than no further pregnancy. Couples with an abnormal parental karyotype were less likely to have a miscarriage than no further pregnancy (RRR: 0.09 (95% CI: 0.01-0.79)). Including successive pregnancies conceived over the study period, the overall livebirth rate was 63% (n = 466/742), but this was reduced to 44% in women aged ≥40 years and 54% in women with infertility. This work covers 13 years; however, those included in the later years have a shorter follow-up time. Although electronic health records have improved data availability, data collection in this cohort remains hampered by the absence of a formal booking visit for women presenting with miscarriage and a national miscarriage database or register. Our findings are largely reassuring as most women with RM and hoping to conceive achieved a livebirth. In addition to older age, smoking and parental balanced translocations were associated with a reduced likelihood of further pregnancy. No investigation or treatment was associated with pregnancy outcome, reiterating the importance of the supportive aspects of care for women and their partners after RM and counselling regarding individual risk factors. This contributes to the limited international data on the investigative findings and treatment of women with RM. The high rate of prescribed medications merits greater scrutiny, in conjunction with other pregnancy outcomes, and reiterates the need for a national guideline on RM. L.A.L. is a PhD scholar funded through the Pregnancy Loss Research Group, Department of Obstetrics and Gynaecology, University College Cork. M.H. and C.F. are Postdoctoral Researchers on a project funded by the Health Research Board Ireland [ILP-HSR-2019-011] and led by K.O.D., titled: 'Study of the impact of dedicated recurrent miscarriage clinics in the Republic of Ireland'. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. The authors have no conflicts of interests to declare. N/A.

Genetic causes of early miscarriage in patients after assisted reproductive technologies

Objective. To analyze the frequency of occurrence of a pathological karyotype during pregnancy that occurred naturally and through the use of ART. Miscarriage (MC) is a significant problem for the medical community. There is a number of factors affecting the process of gestation. Currently, there is an opinion about the impact of assisted reproductive technologies (ART) methods on the fetal karyotype and an increase in the risk for MC when they are used.&#x0D; Materials and methods. The study used the data of karyotyping of abortive material from 256 women diagnosed the non-developing pregnancy with indication of age and gestational age at the time of termination of pregnancy for the period from 2018 to 2020, provided by JSC "Center for Family Medicine" in Yekaterinburg. In the course of the work, a statistical analysis of the frequency of occurrence of pathological karyotypes in different groups, identified on the basis of the method of pregnancy, was carried out.&#x0D; Results. To a greater extent, the problem of MC associated with a pathological fetal karyotype, occurs during natural pregnancy. This may be due to the lack of pregravid preparation. In addition, during ART, especially with the use of donor material, the cells without karyotype abnormalities are selected. The problem of MC after the use of ART may be associated with other somatic or functional risk factors.&#x0D; Conclusions. The available data on the possible genetic causes of early MC draw attention to the need for preimplantation genetic testing to make a timely diagnosis of fetal chromosomal abnormalities. In order to improve the reproductive health of the population, such method as a pregnant woman genetic passport can be proposed.

Does ICSI outcome in obstructive azoospermia differ according to the origin of retrieved spermatozoa or the cause of epididymal obstruction? A comparative study

PurposeTo determine whether ICSI outcomes are affected by sperm source or genital tract inflammatory status.MethodsA retrospective cohort study was conducted in all consecutive obstructive azoospermia patients who underwent testicular sperm aspiration (TESA) or percutaneous epididymal sperm aspiration (PESA) and ICSI between February 1, 2017, and December 31, 2020. Couples were excluded if they were diagnosed with monogenic disease, abnormal karyotype, or had female uterine malformation. The primary objective was to determine whether ICSI outcomes are affected by the use of testicular or epididymal spermatozoa, and the secondary objective was to explore the effect of granulocyte elastase on ICSI outcomes using epididymal spermatozoa.ResultsCompared with TESA, inflammatory and non-inflammatory PESA patients exhibited a better high-quality embryo rate, with significant differences among the three groups (49.43 vs. 55.39% and 56.03%; odds ratio, 6.345 and 6.631; 95% confidence interval, 0.340–12.350, and 1.712–11.550; P = 0.038 and P = 0.008, respectively). The fertilization rate, clinical pregnancy rate, live birth delivery rate, and congenital anomaly birth rate were similar in patients who underwent TESA or PESA (with or without inflammation).ConclusionsThe high-quality embryo rate in PESA patients was higher than that in TESA patients. After successful pregnancy, ICSI outcomes did not differ between patients with obstructive azoospermia who experienced TESA or PESA and those with or without genital tract inflammation.

Investigation of the relationship between reproductive disorders and chromosomal abnormalities in a large-scale, single-center 10-year retrospective study

ObjectiveChromosomal changes are an important cause of reproductive disorders. This study investigated the chromosomal changes and prevalence of pathologies in individuals admitted to our Genetic Evaluation Center over a 10-year period due to a reproductive disorder. Materials & MethodsThe chromosomal findings of 4345 individuals with reproductive disorders who applied to our Genetic Evaluation Center at Akdeniz University in Antalya, Turkey between 2011 and 2021 were retrospectively evaluated. ResultsIn this study, an abnormal karyotype was found in a total of 138 individuals (87 males and 51 females). Although the incidence of this abnormal karyotype varied among the diseases in the reproductive disorder subgroups, it was most frequently seen in azoospermia (17.0%). Of the 138 abnormalities, 75 were numerical and 54 were structural. The remaining 9 abnormalities consisted of 6 sex reversals and 3 patients with both numerical and structural anomalies. Additionally, the X chromosome was the chromosome most frequently involved in these abnormalities, being observed in 40.6% of patients. ConclusionThis 10-year, single-center study involved one of the largest case series in the literature to investigate the subtypes of reproductive disorders and their chromosomal relationship. Although the importance of chromosome analysis has been deemphasized, it is still recommended for use by the guidelines and, as the results of this study demonstrate, is still a highly effective method in the investigation of reproductive disorders. Furthermore, chromosome analysis of individuals diagnosed with a reproductive disorder is also very important in the practice of the increasingly utilized preimplantation genetic diagnosis (PGD).

Prenatal diagnosis of cystic hygroma cases in a tertiary centre and retrospective analysis of pregnancy results

The aim of this study is to retrospectively examine invasive diagnostic methods, structural anomalies accompanying cystic hygroma, and pregnancy outcomes in cystic hygroma cases admitted to a tertiary centre. The population of the study consisted of 29 live foetuses with cystic hygroma in the foetal neck only in the first or second trimester. In the study, pregnant women who applied to our centre were included. Amniocentesis or chorionic villus sampling was performed for genetic analysis according to the weeks of the pregnant women who were diagnosed with cystic hygroma by ultrasound examination by two clinicians experienced in foetal anomaly. Of the pregnant women included in the study, 10 had normal karyotype, 12 had abnormal karyotype and 13 had structural abnormality. It is very important to provide genetic counselling to the families of foetuses with cystic hygroma with a multidisciplinary team approach consisting of neonatologists, paediatric surgeons and experienced sonographers. Implications for rehabilitation What is already known on this subject? Cystic hygroma, also known as cystic lymphangioma, is a congenital cystic malformation often seen in the first trimester, which occurs in the foetal neck due to the failure of the connections between the cervical lymphatic vessels and the jugular venous system to develop normally. Cystic hygroma may be isolated, but highly associated with foetal aneuploidy, hydrops fetalis, abnormal foetal nuchal translucency. What do the results of this study add? Invasive prenatal diagnostic tests (CVS or amniocentesis) should be performed in all patients with cystic hygroma, as cystic hygromas can be diagnosed by first trimester foetal genetic sonogram screening and are largely accompanied by chromosomal abnormalities. What are the implications of these findings for clinical practice and/or further research? In foetuses with cystic hygroma, foetal karyotyping, detailed sonography and their documentation, genetic counselling is important to families of cystic hygroma foetuses with a multidisciplinary team approach consisting of neonatologists, paediatric surgeons and maternal foetal medicine specialists, since there is a high risk for aneuploidy and foetal malformation.

Cytogenetics and Molecular Genetics in Pediatric Acute Lymphoblastic Leukemia (ALL) and Its Correlation with Induction Outcomes.

Arathi SrinivasanAims The aim was to study cytogenetics and molecular genetic profile in pediatric B-acute lymphoblastic leukemia (ALL) and correlate it with induction outcomes. Subjects and Methods A retrospective study of cytogenetics and molecular genetics of 98 children with B-cell ALL from January 2013 to May 2018 was done. Cytogenetics and molecular genetics were done in the bone marrow using multiplex reverse transcription polymerase chain reaction and G-banded karyotyping, respectively. Minimal residual disease (MRD) assessment was done at the end of induction by flowcytometry. Results Of the 98 children, 83 (84.6%) had evaluable cytogenetics, with 11 (13.25%) being abnormal karyotypes. Of the 11 abnormal karyotypes, seven children (8.4%) had hyperdiploidy, one had hypodiploidy, and three had miscellaneous findings. In molecular genetics, TEL-AML1 (ETV6/RUNX1)[t(12;21)] was the most common fusion gene abnormality (12.2% [12/98]), followed by E2A-PBX1 [t(1;19)] (5%), BCR/ABL1 [t(9;22)] (3%), and MLL-AF4 [t(4;11)] (1%). All the 98 children attained morphologic remission at the end of induction. All children with hyperdiploidy (7/7) attained remission and MRD negativity, but one expired during maintenance chemotherapy of disseminated tuberculosis. The child with hypodiploidy was MRD-positive. Three (25%) children with t (12;21) were MRD-positive. All children with Ph + ALL, t(1:19), and t(4;11) were MRD-negative. Fifty-two children had no detected abnormalities, six of whom had MRD positivity (11.5%). Conclusion Cytogenetic and molecular genetic subgrouping prognosticates ALL outcomes. Although 25% of TEL-AML + children had MRD positivity, larger studies are required to validate the same. End-of-induction MRD outcomes did not correlate with chromosomal aberrations.

Microdeleciones del gen AZF en varones azoospérmicos-oligozoospérmicos

Background and objectiveThe presence of microdeletions in the Y-chromosome azoospermia factor (AZF) region (YCMs) is considered the most frequent genetic cause of male infertility along with Klinefelter syndrome. The objective of this study was to investigate the frequencies and type of YCMs in infertile men in Aragon and to analyze the relationship between sex hormones, sperm count and microdeletions in them. Patients and methodsRetrospective descriptive study of 644 men who during 2006–2019 were screened for YCMs using YChromStrip (Operón, Spain) by PCR+reverse hybridization, spermiogram, karyotype and quantification of sex hormones. ResultsThe frequency of YCMs was 3.88% (25/644), not being detected in any patient with mild or normospermic oligozoospermia, that is, in sperm counts higher than 5×106/mL. The group of azoospermic patients was the one that presented a higher frequency of YCMs (14.58%, 14/96). Deletions in the AZFc region were the most frequent (68%). 20% (5/25) of patients with YCMs also presented some type of karyotype abnormality that included aneuploidies, deletions, duplications and/or translocations. Sperm count was significantly lower and FSH and LH concentrations significantly higher in the group of patients with YCMs. ConclusionsYCMs screening is a key test in the diagnostic approach to male infertility. Obtaining an adequate result allows choosing suitable assisted reproduction techniques, preventing unnecessary treatments and the transmission of genetic defects to offspring.

Marrow changes and reduced proliferative capacity of mesenchymal stromal cells from patients with “no-option” critical limb ischemia; observations on feasibility of the autologous approach from a clinical trial

Background aimsApproximately 1 in 3 patients with critical limb ischemia (CLI) are not suitable for surgical or endovascular revascularization. Those “no-option” patients are at high risk of amputation and death. Autologous bone marrow mesenchymal stromal cells (MSCs) may provide a limb salvage option. In this study, bone marrow characteristics and expansion potentials of CLI-derived MSCs produced during a phase 1b clinical trial were compared with young healthy donor MSCs to determine the feasibility of an autologous approach. Cells were produced under Good Manufacturing Practice conditions and underwent appropriate release testing. MethodsFive bone marrow aspirates derived from patients with CLI were compared with six young healthy donor marrows in terms of number of colony-forming units–fibroblast (CFUF) and mononuclear cells. The mean population doubling times and final cell yields were used to evaluate expansion potential. The effect of increasing the volume of marrow on the CFUF count and final cell yield was evaluated by comparing 5 CLI-derived MSCs batches produced from a targeted 30 mL of marrow aspirate to five batches produced from a targeted 100 mL of marrow. ResultsCLI-derived marrow aspirate showed significantly lower numbers of mononuclear cells with no difference in the number of CFUFs when compared with healthy donors’ marrow aspirate. CLI-derived MSCs showed a significantly longer population doubling time and reduced final cell yield compared with young healthy donors' MSCs. The poor growth kinetics of CLI MSCs were not mitigated by increasing the bone marrow aspirate from 30 to 100 mL. ConclusionsIn addition to the previously reported karyotype abnormalities in MSCs isolated from patients with CLI, but not in cells from healthy donors, the feasibility of autologous transplantation of bone marrow MSCs for patients with no-option CLI is further limited by the increased expansion time and the reduced cell yield.

Analysis of clinical features of multiple myeloma with t(8;14)(q24;q32)

Objective: To investigate the clinical manifestations and prognosis of multiple myeloma (MM) patients with t(8;14)(q24;q32). Methods: The clinical data of MM patients with G-banding results from 2004 to 2009 in Hematology Department of People's Hospital of Peking University were retrospectively analyzed. The general data, M protein related examination, cytogenetics data, therapeutic regimen and response evaluation of MM patients with t(8;14)(q24;q32) were collected. Results: Of all newly diagnosed multiple myeloma patients, the number of patients who had G-banding results was 940, among which 265 had abnormal karyotype in G-banding, accounting for 28.19%. The incidence of t(8;14)(q24;q32) detected by G-banding in MM patients was 0.85%(8/940), t(8;14)(q24;q32) accounted for 3.02%(8/265) of all choromosome abnormalities detected by G-banding. Seven of eight patients were male with a median age of 63.5(56-76) and the immunoglobulin sub-types seven in eight patients were lambda. All eight patients had DS stage Ⅲ at the time of initial diagnosis. FISH detection of these eight patients showed six patients(75%) with 1q21 amplification, and five patients(62.5%) with G-banding results showed abnormal chromosome 1. Among the eight patients, the number of patients reached complete response ,very good response and partial response were separately four, one and two, and the overall response rate(ORR) was 87.5%. After the median follow-up 35 months(23-65months), 2 patients died, and the OS of the dead patients exceeded 5 years. Conclusions: Patients with t(8;14)(q24;q32) accounted for 0.85% of the total who have the results of G banding in our hospital. Of our 8 patients, the light chain sub-type Lambda was more than Kappa, the patients were more common in males, accompanied by 1q21 amplification and chromosome 1 abnormality. The tumor load was high at the time of diagnosis, but the overall response to treatment was fair.