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The genomes of living lungfishes can inform on the molecular-developmental basis of the Devonian sarcopterygian fish-tetrapod transition. We de novo sequenced the genomes of the African (Protopterus annectens) and South American lungfishes (Lepidosiren paradoxa). The Lepidosiren genome (about 91 Gb, roughly 30 times the human genome) is the largest animal genome sequenced so far and more than twice the size of the Australian (Neoceratodus forsteri)1 and African2 lungfishes owing to enlarged intergenic regions and introns with high repeat content (about 90%). All lungfish genomes continue to expand as some transposable elements (TEs) are still active today. In particular, Lepidosiren's genome grew extremely fast during the past 100 million years (Myr), adding the equivalent of one human genome every 10 Myr. This massive genome expansion seems to be related to a reduction of PIWI-interacting RNAs and C2H2 zinc-finger and Krüppel-associated box (KRAB)-domain protein genes that suppress TE expansions. Although TE abundance facilitates chromosomal rearrangements, lungfish chromosomes still conservatively reflect the ur-tetrapod karyotype. Neoceratodus' limb-like fins still resemble those of their extinct relatives and remained phenotypically static for about 100 Myr. We show that the secondary loss of limb-like appendages in the Lepidosiren-Protopterus ancestor was probably due to loss of sonic hedgehog limb-specific enhancers.

OBJECTIVES: Determine the incremental diagnostic yield of prenatal exome sequencing (pES) over chromosome microarray (CMA) or G-banding karyotype in fetuses with central nervous system (CNS) abnormalities. METHODS: Data were collected via electronic searches from January 2010 to April 2022 in MEDLINE, Cochrane, Web of Science and EMBASE. The NHS England prenatal exome cohort was also included. Incremental yield was calculated as a pooled value using a random-effects model. RESULTS: Thirty studies were included (n = 1583 cases). The incremental yield with pES for any CNS anomaly was 32% [95%CI 27%-36%; I2 = 72%]. Subgroup analysis revealed apparent incremental yields in; (a) isolated CNS anomalies; 27% [95%CI 19%-34%; I2 = 74%]; (b) single CNS anomaly; 16% [95% CI 10%-23%; I2 = 41%]; (c) more than one CNS anomaly; 31% [95% Cl 21%-40%; I2 = 56%]; and (d) the anatomical subtype with the most optimal yield was Type 1 malformation of cortical development, related to abnormal cell proliferation or apoptosis, incorporating microcephalies, megalencephalies and dysplasia; 40% (22%-57%; I2 = 68%). The commonest syndromes in isolated cases were Lissencephaly 3 and X-linked hydrocephalus. CONCLUSIONS: Prenatal exome sequencing provides a high incremental diagnostic yield in fetuses with CNS abnormalities with optimal yields in cases with multiple CNS anomalies, particularly those affecting the midline, posterior fossa and cortex.

This study aims to describe the indications, complications, yield, and safety of amniocentesis beyond 24 weeks for prenatal diagnostic procedures along with the associated maternal and fetal outcomes.A retrospective analysis was conducted on 60 pregnant women (with 61 fetuses) who underwent amniocentesis at or beyond 24 weeks from March 2021 to June 2023 at a tertiary care referral center. Data was collected from medical records and individual patient followups. Descriptive data was collected on patient demographics, amniocentesis indications, and the test results. The other outcomes analyzed were the procedure-related complications and pregnancy outcomes.The mean gestational age at time of the procedure was 254/7 (241/7–331/7). The most common indication for late amniocentesis was abnormal sonographic findings (44/61, 72.13 %), with structural anomalies being the commonest (21/61, 34.44 %). There were no complications related to the procedure. Of the 60 women, 88.3 % (53/60) continued their pregnancies, while 11.66 % (7/60) opted for termination of pregnancy, and two patients had intrauterine fetal demise (2/61, 3.27 %). Genetic testing revealed abnormalities in 6.55 % (4/61) of cases. Of the 51 pregnancies, 39 delivered vaginally (76.47 %; 39/51) and 12 (23.52 %; 12/51) required caesarean sections. There were five neonatal and infant deaths due to heart defects (2), metabolic syndrome, congenital diaphragmatic hernia, and non-immune hydrops, respectively.Amniocentesis, done at a later gestation, is a safe and an effective tool for prenatal diagnosis and provides an opportunity to make a genetic diagnosis and further counseling. [ABSTRACT FROM AUTHOR]

Vanishing twin syndrome (VTS) refers to the spontaneous reduction of a fetus during multiple pregnancies, resulting in its invisibility during gestation. Vanishing twin syndrome is commonly identified through ultrasonographic examination in early pregnancy; nonetheless, the genetic causes of VTS are rarely detected. This report aimed to investigate the feasibility of genetic testing for VTS using low-coverage whole-genome sequencing and short tandem repeat (STR) analysis. We report a 39-year-old woman who underwent in vitro fertilization and conceived dichorionic diamniotic twins. The heartbeat of 1 fetus ceased at 11 weeks, leading to a diagnosis of VTS, whereas the surviving fetus developed normally and was delivered at full term. Placental examination revealed a rudimentary gestational sac adjacent to the placenta that correlated with the vanished fetus. Chorionic tissues were collected from the placenta of the live-born infant and the rudimentary gestational sac that was considered to have originated from the vanished fetus. Genetic testing of chorionic tissues from the gestational sac and placenta showed that both fetuses were monozygotic twins with a normal 46, XY male karyotype. However, a triallelic pattern at the Penta E locus was identified in the vanished fetus on STR analysis, which was a unique genetic characteristic. This report highlights the feasibility of genetic testing for VTS, despite the unclear relationship between the identified genetic pattern and the vanished fetus. [ABSTRACT FROM AUTHOR]

Cell immortalization has an important role in scientific research, as well as increasing significance in the context of cell therapy and biotechnology. Over the years, many immortalized cell lines have been produced using human telomerase reverse transcriptase (hTERT) alone or in a combination with viral oncogenes. Different hTERT-immortalized cells are commercially available, and numerous papers about obtaining immortalized cell lines have also been published. However, no specific list of characteristics that need to be checked to confirm successful immortalization exists. Most researchers evaluate only a few parameters, while different articles contain various opinions on the assessment of these characteristics. Results also vary significantly between different cell types, which have their own traits depending on their origin and functions. In the current paper, we raise these questions and discuss controversial issues concerning currently available testing methods for immortalization evaluation and the value and the limitations of the approaches. In addition, we propose a protocol for evaluation of hTERT immortalization success consisting of the following important steps: the assessment of the proliferation rate and dividing capacity, cell morphology, phenotype, karyotype stability, telomerase activity, the expression of cell-specific markers, and tumorigenicity. To our opinion, the hTERT expression level, telomere length, and senescence-associated β-galactosidase staining are controversial with regard to the implemented methods, so these parameters may be optional. For all the evaluation steps, we recommend to pay attention to the necessity of comparing the traits of the obtained immortalized and parent cells. [ABSTRACT FROM AUTHOR]

Human allogeneic liver-derived progenitor cells (HALPCs) display advanced ability to differentiate into hepatocyte-like cells and exhibit potent immunomodulatory, anti-inflammatory, and anti-fibrotic properties. HALPCs have been successfully manufactured under good manufacturing practice (GMP) and are currently in clinical development. A previous phase 2a trial demonstrated the safety of peripheral intravenous infusions of HALPCs and preliminary evidence of the cells' properties to restore liver function in patients with acute-on-chronic liver failure (ACLF), thus potentially improving their survival. A phase 2b trial is currently ongoing across multiple centers (NCT04229901) to obtain proof-of-concept on efficacy and additional safety. HALPCs are currently manufactured using fetal bovine serum (FBS), which can reveal qualitative and quantitative variations between batches. The use of serum-free medium (SFM) represents an alternative means to overcome this variability while also complying fully with regulations. The aim of this study was to compare current FBS-containing culture conditions with two industry-available GMP-compliant SFMs: StemMACS (Miltenyi Biotec, Bergisch Gladbach, Germany) and PRIME-XV (FUJIFILM Irvine Scientific, Santa Ana, California, USA). The proliferation of HALPCs was significantly stimulated by both SFMs, which shortened both their emergence period and population doubling time. This effect was correlated with a significant improvement in their genetic stability as analyzed by conventional karyotyping. The expression profile (identity and purity) and functionality of HALPCs cultured in SFM were maintained, as demonstrated by flow cytometry and enzyme-linked immunoassay (ELISA), respectively. Their potency, evaluated via prostaglandin E2 (PGE2) secretion, showed a similar effect on CD4+ T-cell proliferation in FBS and SFM conditions. Furthermore, a greater proportion of HALPCs cultured in SFM showed enhanced expression of tissue factor (CD142) compared with the FBS condition. Altogether, SFM conditions enabled consistent HALPC quality to be achieved without altering their expression and functional profiles. [ABSTRACT FROM AUTHOR]

Morphological analyses of the vegetative characteristics of two species of Tradescantia and two species of Commelina grown in polythene bags in the greenhouse were done. Inter-specific variation in vegetative and floral morphology was significant (p<0.5). The root tips were harvested between 7:30 am and 9:00 am, pretreated in 8- hydroxyl-quinoline, and transferred into 3:1 ethanol acetic acid for 24 hours. Root tips were hydrolyzed in 10% hydrochloric acid and subsequently squashed in FLP orcein. The chromosome number was 2n = 12 in Tradescantia spathacea with a Karyotype formula of 10m + 2sm. The chromosome number of Tradescantia pallida was 2n = 24 with a Karyotype formula of 22m +2sm, while the Chromosome number was 2n = 22 in Commelina benghalensis with a karyotype formula of 14m+2sm + 4st. The chromosome number was 2n = 30 in Commelina diffusa with a karyotype formula of 28m + 2sm. The mean chromosome lengths in the two genera ranged from 1- 4µm. A secondary construction satellite was observed in the longest chromosome of Commelina benghalensis. Morphological and cytological data from the two genera showed considerable differences, which would be helpful to the genera's taxonomic separation. [ABSTRACT FROM AUTHOR]

The origin of hexaploid sweetpotato [Ipomoea batatas (L.) Lam.] remains controversial. Comparative karyotype analysis is particularly useful in determining species relationships and the origin of polyploid species. In previous study, we developed a set of oligo probes and identified all chromosomes of Ipomoea nil, a model diploid Ipomoea species. Here, we found that this set of oligo probes could be used to identify all chromosomes of sweetpotato and its wild relatives with different ploidy. Karyotypes based on individually identified chromosomes were established and the number and position of 5S and 35S rDNA loci were determined for these Ipomoea species. Comparison of their karyotypes revealed distinct variations in the karyotypic parameters. Karyological relationships among these species were revealed by principal coordinate analysis (PCoA) based on six quantitative parameters (x, 2n, TCL, MCA, CVCL and CVCI). These results show that I. trifida is the most closely related diploid species to sweetpotato, and other diploid species could be excluded from consideration as its possible diploid ancestor. In addition, our study also provides cytogenetic evidence for the segmental allopolyploid hypothesis of sweetpotato origin. [ABSTRACT FROM AUTHOR]

SUMMARY Lamiales is one of the largest orders of angiosperms with a complex evolutionary history and plays a significant role in human life. However, the polyploidization and chromosome evolution histories within this group remain in mystery. Among Lamiales, Isodon serra (Maxim.) Kudô shines for its abundance of diterpenes, notably tanshinones, long used in East Asia to combat toxicity and inflammation. Yet, the genes driving its biosynthesis and the factors governing its regulation linger in obscurity. Here, we present the telomere‐to‐telomere genome assembly of I. serra and, through gene‐to‐metabolite network analyses, pinpoint the pivotal tanshinone biosynthesis genes and their co‐expressed transcription factors. Particularly, through luciferase (LUC) assays, we speculate that IsMYB‐13 and IsbHLH‐8 may upregulate IsCYP76AH101, which is the key step in the biosynthesis of the tanshinone precursor. Among Lamiales, Oleaceae, Gesneriaceae and Plantaginaceae successively sister to a clade of seven Lamiales families, all sharing a recent whole‐genome duplication (designated as α event). By reconstructing the ancestral Lamiales karyotypes (ALK) and post‐α event (ALKα), we trace chromosomal evolution trajectories across Lamiales species. Notably, one chromosomal fusion is detected from ALK to ALKα, and three shared chromosomal fusion events are detected sequentially from ALKα to I. serra, which fully supports the phylogeny constructed using single‐copy genes. This comprehensive study illuminates the genome evolution and chromosomal dynamics of Lamiales, further enhancing our understanding of the biosynthetic mechanisms underlying the medicinal properties of I. serra. [ABSTRACT FROM AUTHOR]

Background: A variation in the number of chromosomes can lead to chromosomal disorders. These chromosomal aberrations might be related to autosomes or sex chromosomes. The most common chromosomal aberrations that are sex-linked and autosomal are Klinefelter syndrome and Down syndrome, respectively. However, the worldwide occurrence of double chromosomal aneuploidy in a single individual is relatively exceptional event and random meiotic nondisjunction events result in double aneuploidy, which causes trisomy of two distinct chromosomes. The clinical manifestations vary depending on whether aneuploidy is dominant or an amalgam of both. Moreover, double aneuploidy including autosome and sex chromosome is not often documented. In this paper, we document a case of Down–Klinefelter double aneuploidy syndrome in an Indian infant hospitalized to the neonatal care unit. Case presentation: A full-term 9-month-old male infant born to a 36-year-old female with classical clinical signs of Down syndrome like epicanthus, a depressed nasal bridge, a flat face, small ears, an open mouth, thick lower lip vermilion, a protruding tongue, and a short neck was referred for cytogenetic examination, revealing a rare karyotype of 48, XXY, + 21. Conclusion: Down–Klinefelter syndrome is a rare chromosomal anomaly with unique characteristics, often displaying only Down syndrome-like traits at birth. Early diagnosis can be challenging due to the lack of noticeable symptoms until puberty. Early cytogenetic investigation can detect double aneuploidy, improving the affected person's quality of life and educating family members about potential medical and psychosocial difficulties. [ABSTRACT FROM AUTHOR]

SUMMARY As tertiary gene pools of wheat, Aegilops comosa and Ae. caudata contain many excellent genes/traits and gradually become important and noteworthy wild resources for wheat improvement worldwide. However, the lack of molecular markers and cytological probes with good specificity and high sensitivity limits the development and utilization of Triticum aestivum–Ae. comosa (Ta. Aeco)/Ae. caudata (Ta. Aeca) introgression lines. Using specific‐locus amplified fragment sequencing, two Ae. comosa and one Ae. caudata accessions, Chinese Spring, and three Ta. Aeco and Ta. Aeca introgression lines each were sequenced to develop new molecular markers and cytological probes. After strict sequence comparison and verification in different materials, a total of 39 molecular markers specific to three chromosomes in Ae. comosa (nine, seven, and 10 markers for 1M, 2M, and 7M, respectively) and Ae. caudata (two, six, and five markers for 3C, 4C, and 5C, respectively) and 21 fluorescence in situ hybridization (FISH) probes (one centromeric probe with signals specific to the M chromosomes, two centromeric probes with signals in all the tested genomes, and six, eight, and four FISH probes specific to the M, C, and M, C, and U chromosomes, respectively) were successfully exploited. The newly developed molecular markers and cytological probes could be used in karyotype studies, centromere evolutionary analyses of Aegilops, and had the ability to detect the fusion centromeres and small‐fragment translocations in introgression lines. [ABSTRACT FROM AUTHOR]

Introduction: Karyotyping and chromosome microarray analysis (CMA) are the two main prenatal diagnostic techniques currently used for genetic testing. We aimed to evaluate the value of chromosomal karyotyping and CMA for different prenatal indications. Methods: A total of 2084 amniocentesis samples from pregnant women who underwent prenatal diagnosis from 16 to 22 + 6 weeks of gestation between January 2021 and December 2022 were retrospectively collected. The pregnant women were classified according to different prenatal diagnostic indications and underwent CMA and karyotype analysis. Clinical data were collected, and the results of the CMA and karyotype analysis were statistically analyzed to compare the effects of the two diagnostic techniques. Results: The total detection rate of abnormal chromosomes was significantly higher using CMA than karyotype analysis. The detection rate of abnormal chromosomes using CMA was significantly higher than that using karyotyping for ultrasound abnormalities, high-risk serologic screening, adverse pregnancy history, positive noninvasive prenatal test (NIPT) screening, and ultrasound abnormalities combined with adverse pregnancy history indications. Among the fetuses with inconsistent results between the two testing methods, 144 had an abnormal CMA but a normal karyotype, with the highest percentage of pregnant women with ultrasound abnormalities at 38.89% (56/144). CMA had the highest detection rate for structural abnormalities combined with soft-index abnormalities among all ultrasound abnormalities. The highest detection rate of copy number variants in the group of structural abnormalities in a single system was in the genitourinary system (3/29, 10.34%). Conclusion: CMA can improve the detection rate of chromosomal abnormalities in patients with ultrasound abnormalities, high-risk serologic screening, adverse maternal history, positive NIPT screening, and ultrasound abnormalities combined with adverse maternal history and can increase the detection rate of chromosomal abnormalities in karyotypic normality by 6.91% (144/2,084), this result is higher than similar studies. However, karyotype analysis remains advantageous over CMA regarding balanced chromosomal rearrangement and detection of low-level chimeras, and the combination of the two methods is more helpful in improving the detection rate of prenatal chromosomal abnormalities. [ABSTRACT FROM AUTHOR]

Conventional cytogenetic analysis is an important tool for the diagnosis of many hematologic disorders (HD). A karyotype is designed as « complex » when several alterations are detected. However, there is no clear consensus on the exact definition of a complex karyotype (CK), and there is a lack of studies that exclusively analyze CK in the literature. Complex karyotypes were analyzed over a period of 12 years at the Jacques Loiselet Center for Medical Genetics and Genomics (CGGM) at Saint Joseph University in Beirut (USJ) in Lebanon. 255 CK were analyzed with their associated chromosomal abnormalities (CA) detected. Out of 255 patients, 59.22% were males with a mean age of 59 years. The most common anomaly associated with CK was hyperdiploidy with a prevalence of 22.41%, which is different from a previously published study. To our knowledge, this represents the largest series of CK, particularly within the Middle East region. This study underscores the critical role of conventional cytogenetics in detecting CK, ultimately contributing to improved management of HD. Further investigations focusing on CK are needed. [ABSTRACT FROM AUTHOR]

Trait evolution has become a central focus in evolutionary biology, with phylogenetic comparative methods offering a framework to study how and why traits vary among species. Identifying variations in trait evolution rates within phylogenies is important for uncovering the mechanisms behind these differences. Karyotype variation, which is substantial across eukaryotic organisms, plays an essential role in species diversification. This study investigates karyotype variation within the leafcutting ant clade, focusing on chromosome number and morphology. We aim to determine whether karyotypic traits are phylogenetically dependent and how different evolutionary models explain karyotype diversity. Previous models have been insufficient in explaining these variations. To address these gaps, we employ modern phylogenetic methods to assess the impact of chromosomal fissions and fusions on karyotype evolution. By evaluating various evolutionary models—particularly the Brownian motion model, which suggests neutral chromosomal changes—we pursue for the further understanding the mode and tempo of karyotype evolution in ants. Our research examines how shifts in chromosomal change rates contribute to divergence among leafcutting ant species and assesses the role of chromosomal changes in the clade's evolutionary trajectory. Comparative analysis of leafcutting ant ideograms suggests that shared karyotype traits are strongly related to species relationships. This implies that karyotype diversification in leafcutting ants follows a phylogenetic trajectory at varying rates, with differences in karyotype traits reflecting the evolutionary distance between lineages. Particularly, the increase in the chromosome number of Acromyrmex is likely due to fission rearrangements rather than demi or polyploidization. We discuss and provide insights into the mechanisms driving karyotype variation and its implications for leafcutting ant diversification. [ABSTRACT FROM AUTHOR]

Background/objectives: Acute promyelocytic leukemia (APL) is an aggressive subtype of acute myeloid leukemia (AML), characterized by the hallmark translocation t(15;17) resulting in a PML::RARA fusion. Once diagnosed, APL is now considered to be one of the most treatable forms of AML. However, without early detection and treatment, the disease is associated with rapid deterioration and lethal side effects. Methods: We describe a case of diagnostic APL presenting with a normal karyotype, normal RARA break-apart FISH, and unclear, atypical PML/RARA FISH findings. We used optical genome mapping (OGM) to characterize this atypical PML/RARA fusion. Results: OGM allowed for detection of a PML::RARA fusion resulting from a cryptic and complex insertion of PML::RARA into RARA on 17q21.2 whereby a segment of 15q24.1 was inserted into the 17q21.2. The recipient breakpoint of the insertion was at intron 2 of the RARA gene and the donor breakpoint of the insertion was at exon 5/intron 6 of the PML gene. Conclusions: This is the first report of an insertional PML::RARA fusion into the RARA gene on 17q detected by OGM. OGM has demonstrated its utility in a clinical cytogenetics environment, allowing for clearer characterization and diagnosis of various neoplasms. [ABSTRACT FROM AUTHOR]

Objectives We sought to characterize the immunophenotype of acute myeloid leukemia (AML) with CBFB rearrangement and correlate the results with cytogenetic and molecular data. Methods Sixty-one cases of AML with CBFB rearrangement were evaluated. Results The sample population consisted of 33 men and 28 women, with a median age of 49 years. Flow cytometry immunophenotypic analysis showed that myeloblasts were positive for CD34 and CD117 in all cases, and myeloperoxidase was positive in 52 of 55 (95%) cases. The most common abnormalities included decreased CD38 in 90%, increased CD13 in 85%, increased CD123 in 84%, and decreased HLA-DR in 84% of cases. Monocytes were increased, with a mature immunophenotype, and accounted for 23.7% of total cells. Among 60 cases with available karyotype, inv(16)(p13.1q22) was most common in 50 (83%) cases, followed by t(16;16) (p13.1;q22) in 6 (10%). Type A CBFB::MYH11 transcript was most common, detected in 84% of cases. Mutational analysis showed mutations of NRAS in 37%, FLT3 in 25%, and KIT in 24% of cases. Comparing cases with type A vs non–type A transcripts, blasts in type A cases more frequently exhibited CD64 positivity and increased CD13 levels while showing a lower frequency of CD7 and CD56 expression. Trisomy 22 and mutations in KIT, NF1, and TET2 were identified only in cases with type A transcript. Conclusions Myeloblasts of AML with CBFB rearrangement are positive for CD34, CD117, and myeloperoxidase. These neoplasms most frequently carry inv(16)(p13.1q22) and type A fusion transcript. NRAS mutation was the most common mutation. Some immunophenotypic and genetic correlations occurred with different types of transcripts. [ABSTRACT FROM AUTHOR]

Recurrent pregnancy loss (RPL) affects approximately 15–20% of couples. Constitutional chromosomal abnormalities (CA) are an important cause of RPL because the gametes of carriers may have chromosomal imbalance which is lethal to the fetus. Therefore, the identification of CA helps to provide information about likely outcome of pregnancy in carriers. The objective of this study was to determine the frequency and types of CA in RPL. The study group consisted of individuals with RPL seen at a tertiary care center in South India from 2001 to 2019, whose karyotypes were reviewed retrospectively. Cytogenetic analysis was performed on peripheral blood using standard protocols. The number of metaphases analyzed per patient was 20 and extended up to 50 for confirmation of low-level mosaicism. There were 3488 individuals (1663 couples and 162 where only partner was tested of whom 104 were females). The frequency of CA was 3.6% (127/3488) (M:F ratio of 1:2.6). Structural aberrations were seen in 2.5% (90/3488) and numerical aberrations of sex chromosomes in 1.1% (37/3488) of patients. Structural aberrations included 68 balanced translocations including one three-way translocation, seven inversions, one insertion and duplication each, and 14 Robertsonian translocations. There was one patient each with Turner syndrome (TS) and 47,XYY syndrome. We had 37 mosaics, comprising 29 with mosaic TS and 6 with triple X mosaicism and 2 with mosaicism for balanced translocation. The frequency of CA is comparable to the literature. We have described a few less commonly seen aberrations as well as sex chromosomal aneuploidies, almost all of which were due to low-level mosaicism. [ABSTRACT FROM AUTHOR]

Objective It was to verify the association between the definition of sex of rearing and, clinical and cytogenetic features among patients with genital ambiguity referred without a sex assignment. Methods The sample consisted of 133 patients with genital ambiguity seen at a single reference service. These patients did not have a defined social sex at the first consultation and their etiological diagnosis was obtained during follow-up. Results A total of 133 cases were included, 74 of which were reared as males and 59 as females. No correlation was found between the year of birth and the year of the first consultation with the definition of sex of rearing. However, the definition of sex of rearing was associated with age at the first consultation, severity of genital ambiguity, presence of palpable gonad(s), presence of uterus on ultrasound, karyotype, and diagnosis. Palpable gonad(s), more virilized genitalia, absence of a uterus on ultrasound, 46, XY karyotype, or a karyotype with sex chromosome abnormalities emerged as strong predictors for defining male sex. All 77 (58 %) patients over 18 years old had a gender identity in accordance with the sex of rearing; though 9 of 77 (12 %) had homo or bisexual orientation, especially girls with Congenital Adrenal Hyperplasia. Conclusions Clinical and cytogenetic data were strongly associated with the definition of the sex of rearing of children with genital ambiguity referred to a DSD center without sex assignment. Management in a specialized center allows the establishment of a gender identity in accordance with the sex of rearing. [ABSTRACT FROM AUTHOR]

Bitter gourd is a highly nutritious vegetable and important medicinal plant of economic importance. The present study is focused on cytogenetical characterization of 12 accessions of bitter gourd from different parts of India, aiming to differentiate their karyotypes and outline diagnostic features of the chromosomes within each accession's haploid complement. All the accessions possess 2 n = 22 numbers of chromosomes. The chromosomes mainly were metacentric (16‒22 chromosomes), and the presence or absence of sub-metacentric (0‒6 chromosomes) chromosomes. The length of the chromosomes varied from 0.83 to 1.93 μm among the accessions studied. Significant differences were obtained for the seven intra-chromosomal indices and four inter-chromosomal indices among the accessions. Principal component analysis and unweighted pair group method with arithmetic mean study revealed relatively distant positioning of individuals that advocated intraspecific phylogenetic relationships and higher karyoevolutionary affinity in bitter gourd accessions. In the meiotic study, regular meiotic behaviour indicates genetic stability and a stable sexual cycle in different accessions. The percentage of pollen viability of all the studied accessions was very high (89.41–94.11%), and these accessions can be considered to be good pollinators. The results obtained will guide characterizing the elite genotypes, genotypes management and designing effective breeding programmes and crop improvement programmes. [ABSTRACT FROM AUTHOR]

Simple Summary: Chromosome diversification represents a fundamental driver of biological evolution. Here we present and discuss the results of a comparative chromosome analysis on different species of the Malagasy chameleons of the genera Brookesia and Furcifer. We show that the study species are characterized by different karyotypes in terms of chromosome number (2n = 36–22), ratio of micro- and macrochromosomes, and the variable presence of differentiated sex chromosomes. Considering our new data together with those from previous studies, we describe a chromosome evolutionary scenario in the studied taxa. Specifically, while genus Brookesia has a fixed chromosome number (2n = 36) and no differentiated sex chromosomes, which corresponds to the hypothesized ancestral chameleon karyotype, Furcifer shows a high diversity in chromosome number (2n = 20–34), morphology, and the independent raising of simple and multiple sex chromosomes with female heterogamety. The karyotypes in Furcifer with a relatively low chromosome count likely evolved from a chromosome complement similar to that of F. balteatus (2n = 34), mostly via a progressive number of chromosome fusions involving distinct micro- and macrochromosome pairs. Similarly, the diversification of simple and multiple sex chromosome systems occurred in Furcifer via independent (non-homologous) sex chromosome-autosome fusions and heterochromatinization. In this work, we performed a preliminary molecular analysis and a comparative cytogenetic study on 5 different species of Malagasy chameleons of the genus Brookesia (B. superciliaris) and Furcifer (F. balteautus, F. petteri, F. major and F. minor). A DNA barcoding analysis was first carried out on the study samples using a fragment of the mitochondrial gene coding for the cytochrome oxidase subunit 1 (COI) in order to assess the taxonomic identity of the available biological material. Subsequently, we performed on the studied individuals a chromosome analysis with standard karyotyping (5% Giemsa solution at pH 7) and sequential C-banding + Giemsa, + CMA3, and + DAPI. The results obtained indicate that the studied species are characterized by a different chromosome number and a variable heterochromatin content and distribution, with or without differentiated sex chromosomes. In particular, B. superciliaris (2n = 36) and F. balteatus (2n = 34) showed a similar karyotype with 6 macro- and 12–11 microchromosome pairs, without differentiated sex chromosomes. In turn, F. petteri, F. major, and F. minor showed a karyotype with a reduced chromosome number (2n = 22–24) and a differentiated sex chromosome system with female heterogamety (ZZ/ZW). Adding our newly generated data to those available from the literature, we highlight that the remarkable chromosomal diversification of the genus Furcifer was likely driven by non-homologous chromosome fusions, including autosome–autosome, Z–autosome, and W–autosome fusions. The results of this process resulted in a progressive reduction in the chromosome number and partially homologous sex chromosomes of different shapes and sizes. [ABSTRACT FROM AUTHOR]

Objectives: The corpus callosum is the main pathway that connects interhemispheric communication. Agenesis of corpus callosum (ACC) have not consistently detected replicate genetic risk factors, potentially due to Etiological heterogeneity of this trait. This study aimed to retrospectively analyze the molecular basis for the ACC and the potential genotyping-phenotyping association and provide the basis for genetic counselling. Material and methods: Karyotyping and chromosomal microarray analysis were performed for copy number variants. Results: Three cases had 1p36 deletions, two cases had 2q31.2 and 2p16.3 microdeletions, one case had microdeletion of Xq26.3q27.1, five cases involved derived chromosomes due to unbalanced translocations. These cases had variable deletions and duplications with partial overlapping. Phenotypically, besides agenesis of corpus callosum and other brain morphological abnormalities as well as heart abnormalities. Conclusions: ACC may occur alone or be related to other abnormal clinical phenotypes, and its genetic mechanism is very complicated. These results revealed ACC is associated with a variety of chromosomal abnormalities. The findings of the present study expand the genotypes associated with ACC, and further delineation of the genotype-phenotype correlations for ACC. With current applications of chromosome microarray analysis, congenital submicroscopic copy-number variations in fetuses can be detected more effectively. [ABSTRACT FROM AUTHOR]

Zoraptera (also called "angel insects") is one of the most unexplored insect orders. However, it holds promise for understanding the evolution of insect karyotypes and genome organization given its status as an early branching group of Polyneoptera and Pterygota (winged insects) during the Paleozoic. Here, we provide karyotype descriptions of three Zorapteran species: Brazilozoros huxleyi (2n♂; ♀ = 42; 42), B. kukalovae (2n♂; ♀ = 43; 44) and Latinozoros cacaoensis (2n♂; ♀ = 36; 36). These species represent two of the four recently recognized Zorapteran subfamilies. Contrary to an earlier suggestion that Zoraptera has holocentric chromosomes, we found karyotypes that were always monocentric. Interestingly, we detected both X0 (B. kukalovae) and XY (B. huxleyi, L. cacaoensis) sex chromosome systems. In addition to conventional karyotype descriptions, we applied fluorescent in situ hybridization for the first time in Zoraptera to map karyotype distributions of 18S rDNA, histone H3 genes, telomeres and (CAG)n and (GATA)n microsatellites. This study provides a foundation for cytogenetic research in Zoraptera. [ABSTRACT FROM AUTHOR]

Abstract Background A variation in the number of chromosomes can lead to chromosomal disorders. These chromosomal aberrations might be related to autosomes or sex chromosomes. The most common chromosomal aberrations that are sex-linked and autosomal are Klinefelter syndrome and Down syndrome, respectively. However, the worldwide occurrence of double chromosomal aneuploidy in a single individual is relatively exceptional event and random meiotic nondisjunction events result in double aneuploidy, which causes trisomy of two distinct chromosomes. The clinical manifestations vary depending on whether aneuploidy is dominant or an amalgam of both. Moreover, double aneuploidy including autosome and sex chromosome is not often documented. In this paper, we document a case of Down–Klinefelter double aneuploidy syndrome in an Indian infant hospitalized to the neonatal care unit. Case presentation A full-term 9-month-old male infant born to a 36-year-old female with classical clinical signs of Down syndrome like epicanthus, a depressed nasal bridge, a flat face, small ears, an open mouth, thick lower lip vermilion, a protruding tongue, and a short neck was referred for cytogenetic examination, revealing a rare karyotype of 48, XXY, + 21. Conclusion Down–Klinefelter syndrome is a rare chromosomal anomaly with unique characteristics, often displaying only Down syndrome-like traits at birth. Early diagnosis can be challenging due to the lack of noticeable symptoms until puberty. Early cytogenetic investigation can detect double aneuploidy, improving the affected person's quality of life and educating family members about potential medical and psychosocial difficulties.

Synthetic intergeneric amphydiploids and genome-substituted wheat forms are an important source for transferring agronomically valuable genes from wild species into the common wheat (Triticum aestivum L.) genome. They can be used both in academic research and for breeding purposes as an original material for developing wheatalien addition and substitution lines followed by translocation induction with the aid of irradiation or nonhomologous chromosome pairing. The chromosome sets and genome constitutions of allopolyploids are usually verified in early hybrid generations, whereas the subsequent fate of these hybrids remains unknown in most cases. Here we analyze karyotypes of five hexa- (2n = 6x = 42) and octoploid (2n = 8x = 56) amphydiploids of wheat with several species of the Aegilops, Haynaldia, and Hordeum genera, and six genome-substituted wheat–Aegilops forms, which were developed over 40 years ago and have been maintained in different gene banks. The analyses involve C-banding and fluorescence in situ hybridization (FISH) with pAs1 and pSc119.2 probes. We have found that most accessions are cytologically stable except for Avrodes (genome BBAASS, a hexaploid genome-substituted hybrid of wheat and Aegilops speltoides), which segregated with respect to chromosome composition after numerous reproductions. Chromosome analysis has not confirmed the presence of the N genome from Ae. uniaristata Vis. in the genome-substituted hybrid Avrotata. Instead, Avrotata carries the D genome. Our study shows that octoploid hybrids, namely AD 7, AD 7147 undergo more complex genome reorganizations as compared to hexaploids: the chromosome number of two presumably octoploid wheatAegilops hybrids were reduced to the hexaploid level. Genomes of both forms lost seven chromosome pairs, which represented seven homoeologous groups and derived from different parental subgenomes. Thus, each of the resulting hexaploids carries a synthetic/hybrid genome consisting of a unique combination of chromosomes belonging to different parental subgenomes.

Spiders are a hyperdiverse taxon and among the most abundant predators in nearly all terrestrial habitats. Their success is often attributed to key developments in their evolution such as silk and venom production and major apomorphies such as a whole‐genome duplication. Resolving deep relationships within the spider tree of life has been historically challenging, making it difficult to measure the relative importance of these novelties for spider evolution. Whole‐genome data offer an essential resource in these efforts, but also for functional genomic studies. Here, we present de novo assemblies for three spider species: Ryuthela nishihirai (Liphistiidae), a representative of the ancient Mesothelae, the suborder that is sister to all other extant spiders; Uloborus plumipes (Uloboridae), a cribellate orbweaver whose phylogenetic placement is especially challenging; and Cheiracanthium punctorium (Cheiracanthiidae), which represents only the second family to be sequenced in the hyperdiverse Dionycha clade. These genomes fill critical gaps in the spider tree of life. Using these novel genomes along with 25 previously published ones, we examine the evolutionary history of spidroin gene and structural hox cluster diversity. Our assemblies provide critical genomic resources to facilitate deeper investigations into spider evolution. The near chromosome‐level genome of the 'living fossil' R. nishihirai represents an especially important step forward, offering new insights into the origins of spider traits. [ABSTRACT FROM AUTHOR]

Introduction: Disorders of sexual differentiation or differences in sex development (DSD) is a congenital anomaly with abnormal differentiation between internal gonads and external genitalia. We present the clinical, genetic, biochemical, hormonal, and radiological profile of DSD patients who presented at a tertiary care institute in Chhattisgarh during 2 years. Methodology: All patients with DSD and age

Androgen Insensitivity Syndrome (AIS) is a rare X-linked Disorder of Sexual Differentiation (DSD) caused by a mutation in the Androgen Receptor (AR) gene, which is located on the X chromosome (Xq11-q12). It can present with a wide spectrum of phenotypes depending on different mutations of the AR gene. It is classified into mild, partial, and complete AIS. In this series of cases, authors describe patients who presented to tertiary hospitals over several years. Case 1 presented with inguinal masses and primary amenorrhoea, which upon investigations revealed complete AIS. Case 2 was a young child who underwent surgery for an inguinal hernia, and later histopathology and karyotype revealed a diagnosis of complete AIS. Case 3 presented with large adnexal masses to oncology, where intraoperatively absent Mullerian structures, histopathology, and karyotype gave the diagnosis of Complete AIS. Case 4 presented with hirsutism, primary amenorrhoea, and clitoromegaly. Upon investigation, imaging and karyotype with virilising features led to the diagnosis of partial AIS. The absence of Mullerian structures on imaging, chromosomal analysis, and the finding of undescended testicular masses in inguinal regions clinch the diagnosis. These patients should undergo gonadectomy after puberty to decrease the chances of malignancy. Therefore, when investigating the cases of primary amenorrhoea, patients with absent pubic/axillary hair and absent uterus/ovaries should raise suspicion of AIS. Also, any young female child with an inguinal hernia should be investigated for AIS.

Abstract Background White clover (Trifolium repens) is a globally important perennial forage legume. This species also serves as an eco-evolutionary model system for studying within-species chemical defense variation; it features a well-studied polymorphism for cyanogenesis (HCN release following tissue damage), with higher frequencies of cyanogenic plants favored in warmer locations worldwide. Using a newly generated haplotype-resolved genome and two other long-read assemblies, we tested the hypothesis that copy number variants (CNVs) at cyanogenesis genes play a role in the ability of white clover to rapidly adapt to local environments. We also examined questions on subgenome evolution in this recently evolved allotetraploid species and on chromosomal rearrangements in the broader IRLC legume clade. Results Integration of PacBio HiFi, Omni-C, Illumina, and linkage map data yielded a completely de novo genome assembly for white clover (created without a priori sequence assignment to subgenomes). We find that white clover has undergone extensive transposon diversification since its origin but otherwise shows highly conserved genome organization and composition with its diploid progenitors. Unlike some other clover species, its chromosomal structure is conserved with other IRLC legumes. We further find extensive evidence of CNVs at the major cyanogenesis loci; these contribute to quantitative variation in the cyanogenic phenotype and to local adaptation across wild North American populations. Conclusions This work provides a case study documenting the role of CNVs in local adaptation in a plant species, and it highlights the value of pan-genome data for identifying contributions of structural variants to adaptation in nature.

This case report delves into the complexities surrounding Edwards syndrome (Trisomy 18), a chromosomal abnormality stemming from meiotic disjunction. Initially reported in 1960, Edwards syndrome is the second most prevalent autosomal trisomy following Down syndrome, affecting between 1 in 3500 and 1 in 7000 individuals, with a slight male predominance. The majority of cases involve parents under 30 years old. A review of 152 cases demonstrates a myriad of anomalies, including severe intellectual deficits, congenital heart problems and distinct facial features. This report aims to enhance understanding of Edwards syndrome by presenting a comprehensive case study detailing the diagnosis and distinctive clinical features of a newborn with Trisomy 18. This case report helps in understanding the complexity of genetics and clinical manifestations, highlighting the importance of prenatal diagnosis and counseling, multidisciplinary care approaches, parental decision-making, palliative care and the need for further research. The case report underscores the necessity for heightened public awareness and support for affected families. This study contributes to the body of knowledge regarding Trisomy 18, fostering a more compassionate and informed approach to managing this challenging genetic disorder, ultimately aiming to improve the quality of life for those affected by Edwards syndrome.

Various soft markers can be detected in the ultrasonography of foetuses, which can be related to chromosomal abnormalities and increases the risk of abnormalities, or they can be considered as normal variations that can disappear due to the pregnancy progress. There are different tools to detect chromosomal abnormalities like conventional karyotyping, chromosomal microarray analysis (CMA), single nucleotide polymorphism (SNP) array, non-invasive prenatal test (NIPT), and non-invasive prenatal screening (NIPS). Therefore, in the present study, we aim to assess the accuracy of ultrasonic soft markers in the diagnosis of chromosomal abnormalities such as chromosomal structural abnormalities, aneuploidy, and triploidy, especially Trisomy 21 and Trisomy 18. A systemic literature search was performed using PubMed, Scopus, Google Scholar, and Web of Science. We gathered all articles published before August 2023. We selected English studies such as retrospective and cross-sectional ones that assessed the relationship between ultrasonic soft markers and foetal chromosomal abnormalities. A total of 10 articles with 18,580 cases were included in our systematic review article that assessed the foetal abnormalities and aneuploidies by using conventional karyotyping, SNP array, CMA, and NIPT (or NIPS). Trisomy 21, Trisomy 18, and chromosomal structural abnormalities were the most common abnormalities related to ultrasonic soft markers by karyotyping; however, Trisomy 13, 47, XXY, 45, X, and mosaic chromosomal abnormalities were other abnormalities detected. Results by CMA showed Trisomy 21 and Trisomy 18 as the most common abnormalities in the foetuses also with ultrasonic soft markers, and other abnormalities were pathogenic copy-number variations, Turner (XO), polyploidy, 22q11.2deletion, and Trisomy13, respectively. It was discovered that there is a greater possibility of having pathogenic copy number variations (CNVs) in the groups with multiple ultrasonic soft markers, while foetuses with ultrasonic soft markers have a decreased prevalence of CMA abnormality compared to those who had significant abnormalities or abnormal nuchal translucency. Trisomy 21 was the only abnormality found by NIPT in the groups with 1 and 2 soft markers, while groups with multiple soft markers were all normal. By using SNP array, it was identified that the rate of chromosomal abnormalities such as aneuploidy and triploidy, LOH, and CNVs was lower in the group with a single ultrasonic soft marker compared to the group with structural abnormalities in multiple systems. Trisomy 21, Trisomy 18, and chromosomal structural abnormalities were the most common chromosomal abnormalities that ultrasound soft markers could diagnose. Therefore, it is recommended to employ soft markers besides CMA, SNP array, and NIPS (or NIPT) for greater accuracy in detecting foetus abnormalities.

Chromosome number is a fundamental genomic trait that is often the first recorded characteristic of a genome. Across large clades, a common pattern emerges: many or even most lineages exhibit relative stasis, while a handful of lineages or species exhibit striking variation. Despite recent developments in comparative methods, most of this heterogeneity is still poorly understood. It is essential to understand why some lineages have rapid rates of chromosome number evolution, as it can impact a variety of other traits. Previous research suggests that biased female meiotic drive may shape rates of karyotype evolution in some mammals. However, Carnivora exhibits variation that this female meiotic drive model cannot explain. We hypothesize that variation in effective population size may underlie rate variation in Carnivora. To test this hypothesis, we estimated rates of fusions and fissions while accounting for range size, which we use as a proxy for effective population size. We reason fusions and fissions are deleterious or underdominant and that only in lineages with small range sizes will these changes be able to fix due to genetic drift. In this study, we find that the rates of fusions and fissions are elevated in taxa with small range sizes relative to those with large range sizes. Based on these findings, we conclude that 1) naturally occurring structural mutations that change chromosome number are underdominant or mildly deleterious, and 2) when population sizes are small, structural rearrangements may play an important role in speciation and reduction in gene flow among populations. [ABSTRACT FROM AUTHOR]

Disorders of sex development (DSD) result from intrauterine defects in sex discrimination. The clinical phenotype differs based on the disease type. Cases with ambiguous external genitalia are diagnosed at birth. However, diagnosis of cases with normal-appearing external genitalia may be delayed until puberty. Here, we report a patient with a pelvic mass and a small uterus that was diagnosed by abdominal ultrasound, in addition to the history of primary amenorrhea and physical examination suggested Swyer syndrome, confirmed by genetic karyotyping. Pathological examination of the surgically removed mass revealed dysgerminoma. Until the age of 19, the patient did not have any idea about 46, XY karyotype, and assumed to be a female. The development of dysgerminoma (as a result of the simultaneous presence of gonadal dysgenesis and Y-chromosome) was another challenge that the patient had to deal with. The diagnosis of this patient at an earlier age could have prevented the development of gonadoblastoma, by removal of the streak gonads. By the presentation of this case, we intend to increase the physician's awareness about DSDs; earlier diagnosis may help the patient deal with her disease better and reduce the risk of further complications. [ABSTRACT FROM AUTHOR]

Background: White clover (Trifolium repens) is a globally important perennial forage legume. This species also serves as an eco-evolutionary model system for studying within-species chemical defense variation; it features a well-studied polymorphism for cyanogenesis (HCN release following tissue damage), with higher frequencies of cyanogenic plants favored in warmer locations worldwide. Using a newly generated haplotype-resolved genome and two other long-read assemblies, we tested the hypothesis that copy number variants (CNVs) at cyanogenesis genes play a role in the ability of white clover to rapidly adapt to local environments. We also examined questions on subgenome evolution in this recently evolved allotetraploid species and on chromosomal rearrangements in the broader IRLC legume clade. Results: Integration of PacBio HiFi, Omni-C, Illumina, and linkage map data yielded a completely de novo genome assembly for white clover (created without a priori sequence assignment to subgenomes). We find that white clover has undergone extensive transposon diversification since its origin but otherwise shows highly conserved genome organization and composition with its diploid progenitors. Unlike some other clover species, its chromosomal structure is conserved with other IRLC legumes. We further find extensive evidence of CNVs at the major cyanogenesis loci; these contribute to quantitative variation in the cyanogenic phenotype and to local adaptation across wild North American populations. Conclusions: This work provides a case study documenting the role of CNVs in local adaptation in a plant species, and it highlights the value of pan-genome data for identifying contributions of structural variants to adaptation in nature. [ABSTRACT FROM AUTHOR]

Androgen Insensitivity Syndrome (AIS) is a rare X-linked Disorder of Sexual Differentiation (DSD) caused by a mutation in the Androgen Receptor (AR) gene, which is located on the X chromosome (Xq11-q12). It can present with a wide spectrum of phenotypes depending on different mutations of the AR gene. It is classified into mild, partial, and complete AIS. In this series of cases, authors describe patients who presented to tertiary hospitals over several years. Case 1 presented with inguinal masses and primary amenorrhoea, which upon investigations revealed complete AIS. Case 2 was a young child who underwent surgery for an inguinal hernia, and later histopathology and karyotype revealed a diagnosis of complete AIS. Case 3 presented with large adnexal masses to oncology, where intraoperatively absent Mullerian structures, histopathology, and karyotype gave the diagnosis of Complete AIS. Case 4 presented with hirsutism, primary amenorrhoea, and clitoromegaly. Upon investigation, imaging and karyotype with virilising features led to the diagnosis of partial AIS. The absence of Mullerian structures on imaging, chromosomal analysis, and the finding of undescended testicular masses in inguinal regions clinch the diagnosis. These patients should undergo gonadectomy after puberty to decrease the chances of malignancy. Therefore, when investigating the cases of primary amenorrhoea, patients with absent pubic/axillary hair and absent uterus/ovaries should raise suspicion of AIS. Also, any young female child with an inguinal hernia should be investigated for AIS. [ABSTRACT FROM AUTHOR]

Objectives: Evaluate the prevalence of genetic factors in a large population of infertile subjects and define the seminological, hormonal, and ultrasonographic features for each alteration. Methods: This single-center retrospective study included male partners of infertile couples undergoing genetic investigations due to oligozoospermia or azoospermia evaluated from January 2012 to January 2022. The genetic investigations consist of karyotype, CFTR gene mutations plus variant of the IVS8-5T polymorphic trait, Y chromosome microdeletion, and Next Generation Sequencing panel to analyze genes implicated in congenital hypogonadotropic hypogonadism (CHH). Results: Overall, 15.4% (72/466) of patients received a diagnosis of genetic cause of infertility. Specifically, 23 patients (31.9%) harbor mutations in the CFTR gene, 22 (30.6%) have a 47, XXY karyotype, 14 (19.4%) patients show a Y chromosome microdeletion, 7 (9.7%) have structural chromosomal anomalies, and 6 (8.3%) have CHH. Overall, 80.6% of patients were azoospermic and 19.4% oligozoospermic (sperm concentration 3.5 ± 3.8 million/mL). Almost all patients presented hormonal alterations related to the specific genotype, while the main ultrasound alterations were testicular hypoplasia, calcifications/microcalcifications, and enlarged/hyperechoic epididymis. Conclusions: The prevalence of genetic abnormalities in males of infertile couples was 15.4% in our Center. CFTR gene disease-causing variants resulted in more frequent, with various clinical features, highlighting the complexity and heterogeneity of the presentation. Other investigations are needed to understand if conditions like ring chromosomes and other translocations are related to infertility or are incidental factors. [ABSTRACT FROM AUTHOR]

Plebeia is a taxonomically complex genus of stingless bees characterized by remarkable interspecific morphological similarity. On the other hand, cytogenetic data have been useful to resolve taxonomic uncertainties and to infer the diversification processes. Therefore, the goal of this study was to provide a karyotypic analyses in five nominal taxa of Plebeia from northeastern Brazil: P. droryana Friese (Természetrajzi Füzetek kiadja a Magyar nemzeti Muzeum 23:381–394, 1900), P. cf. mosquito Smith 1863, Plebeia aff. droryana 1, Plebeia aff. droryana 2, and Plebeia aff. fravocincta based on traditional and molecular cytogenetic methods to test their applicability in cytotaxonomy and to understand the karyoevolutionary processes in these bees. All species shared a chromosomal number of 2n = 34, except for some individuals of Plebeia aff. flavocincta that presented up to two small and heterochromatic B chromosomes. Species-specific karyotype formulae were observed while the heterochromatin segments were mainly distributed on short arms and pericentromeric regions of most chromosomes, including subtle C-banding differences in each species. The 18S rDNA sites were terminally located on pair 3 of P. droryana, P. cf. mosquito, Plebeia aff. droryana 2, and Plebeia aff. droryana 1, and on the first metacentric pair in Plebeia aff. flavocincta. These data reinforced the efficiency of cytogenetics as an informative tool to discriminate Plebeia species. The present data suggest that structural rearrangements such as inversions and duplications/deletions of heterochromatin and ribosomal clusters are the main drivers of karyotypic evolution in Plebeia. [ABSTRACT FROM AUTHOR]

Cytogenetic information about the product of conception (POC) is important to determine the presence of recurrent chromosomal abnormalities that are an indication for preimplantation genetic testing for aneuploidy or structural rearrangements. Although microscopic examination by G-staining has long been used for such an evaluation, detection failures are relatively common with this method, due to cell-culture-related issues. The utility of low-coverage whole-genome sequencing (lcWGS) using short-read next-generation sequencing (NGS) has been highlighted recently as an alternative cytogenomic approach for POC analysis. We, here, performed comparative analysis of two NGS-based protocols for this purpose based on different short-read sequencers (the Illumina VeriSeq system using a MiSeq sequencer and the Thermo Fisher ReproSeq system using an Ion S5 sequencer). The cytogenomic diagnosis obtained with each NGS method was equivalent in each of 20 POC samples analyzed. Notably, X chromosome sequence reads were reduced in some female samples with both systems. The possibility of low-level mosaicism for monosomy X as an explanation for this was excluded by FISH analysis. Additional data from samples with various degrees of X chromosome aneuploidy suggested that it was a technical artifact related to X chromosome inactivation. Indeed, subsequent nanopore sequencing indicated that the DNA in the samples showing the artifact was predominantly unmethylated. Our current findings indicate that although X chromosome data must be interpreted with caution, both the systems we tested for NGS-based lcWGS are useful alternatives for the karyotyping of POC samples. [ABSTRACT FROM AUTHOR]

Six medicinally important species of Acanthaceae family, namely Asystasia gangetica, Barleria cristata, B. prionitis, Ruellia simplex, Thunbergia erecta and T. mysorensis were cytogenetically investigated to evaluate the karyotype diversity and chromosomal relationship. The somatic chromosome numbers and the karyotype formulae of these species were 2n = 2x = 24 (22m + 2sm) for A. gangetica, 2n = 4x = 40 (38m + 2sm) for B. cristata, 2n = 4x = 40 (36m + 4sm) for B. prionitis, 2n = 2x = 36 (34m + 2sm) for R. simplex, 2n = 4x + 6 = 62 (56m + 6sm) for T. erecta and 2n = 2x = 34 (34m) for T. mysorensis. Considering the karyotype asymmetry parameters, T. erecta, R. simplex, B. cristata and B. prionitis were shown to be cytogenetially more advanced than A. gangetica and T. mysorensis. Three categories of karyotypes were observed such as 1B type in B. cristata, B. prionitis, R. simplex and T. mysorensis, whereas the most primitive one 1A in A. gangetica and the most advanced karyotype 2B in T. erecta. Three new reports of somatic chromosome count yielded in case of A. gangetica, R. simplex and T. mysorensis which might have resulted from aneuploidy or allopolyploidy. The cytological data presented for the six species demonstrated significant variation and distinctiveness from one another, assisting in the evaluation of the karyological diversity that aided in the understanding of evolutionary changes. [ABSTRACT FROM AUTHOR]

Premise: Cytotaxonomy employs chromosome visualization to study organismal relationships and evolution. Despite the critical value of cytogenetic data, cytotypes are lacking for many plant groups. Here, we present an improved approach for visualizing mitotic chromosomes in ferns, a key lineage of land plants, using the dividing cells of unfurling croziers (fiddleheads). Methods and Results: Our modified mitotic chromosome preparation incorporates a brief pectinase–cellulase pretreatment, as well as colchicine fixation and the Feulgen reaction to improve the staining and separation of mitotic chromosomes. To demonstrate this easy and efficient assessment, we determined the sporophytic (2n) chromosome number for three fern species: Cheilanthes mollis (2n = 60), Cheilanthes hypoleuca (2n = 120), and Nephrolepis cordifolia (2n = 82). Conclusions: The new method presented here improves visualizations of mitotic chromosomes from the dividing nuclei of young fern croziers. Fiddleheads are widely accessible in nature and in living collections worldwide, and this modified approach increases their suitability for fern cytotaxonomic studies. [ABSTRACT FROM AUTHOR]

Purpose: The main purpose of this study is to compare the validity of transcervical embryoscopy method with standard uterine evacuation method in detecting more accurate karyotypes in miscarriages below tenth week of pregnancy. Additionally, the frequency and distribution of fetal morphological abnormality were evaluated. Methods: A prospective study was carried out at the Gazi University Faculty of Medicine, Department of Obstetrics and Gynecology. Patients with missed abortions between sixth and tenth gestational weeks were included in the study group, and fetal morphological examination and direct embryonic biopsy were performed by transcervical embryoscopy. The control group consisted of patients who experienced miscarriage and genetic material obtained from routine uterine evacuation between February and October 2023. Result: A total of 60 patients in the study group and 189 patients in the control group were evaluated. The median ages, previous miscarriage numbers, median gravida numbers, and median gestational weeks were comparable between groups. Chromosomal abnormality was detected in 24 (42.8%) and 52 embryos (29.9%) in the study and control groups, respectively (p = 0.004). Culture failure rates were 6.6% (n = 4) and 7.9% (n = 15) in the study and control groups, respectively. In the study group, 12 embryos had a morphological abnormality in which 6 of them had normal karyotype. Conclusion: Direct embryonic biopsy with transcervical embryoscopy is an effective method to exclude maternal decidual cell contamination and placental mosaicism in miscarriages for karyotype analysis. In addition, detecting anomalies in morphology might contribute our understanding in the process of miscarriages which arises independent from structural/numerical chromosomal abnormalities. [ABSTRACT FROM AUTHOR]

Schoenoplectus tabernaemontani (C. C. Gmelin) Palla is a typical macrophyte in diverse wetland ecosystems. This species holds great potential in decontamination applications and carbon sequestration. Previous studies have shown that this species may have experienced recent polyploidization. This would make S. tabernaemontani a unique model to study the processes and consequences of whole-genome duplications in the context of the well-documented holocentric chromosomes and dysploidy events in Cyperaceae. However, the inference was not completely solid because it lacked homology information that is essential to ascertain polyploidy. We present here the first chromosome-level genome assembly for S. tabernaemontani. By combining Oxford Nanopore Technologies (ONT) long reads and Illumina short reads, plus chromatin conformation via the Hi-C method, we assembled a genome spanning 507.96 Mb, with 99.43% of Hi-C data accurately mapped to the assembly. The assembly contig N50 value was 3.62 Mb. The overall BUSCO score was 94.40%. About 68.94% of the genome was comprised of repetitive elements. A total of 36,994 protein-coding genes were predicted and annotated. Long terminal repeat retrotransposons accounted for ∼26.99% of the genome, surpassing the content observed in most sequenced Cyperid genomes. Our well-supported haploid assembly comprised 21 pseudochromosomes, each harboring putative holocentric centromeres. Our findings corroborated a karyotype of 2 n = 2 X = 42. We also confirmed a recent whole-genome duplication occurring after the divergence between Schoenoplecteae and Bolboschoeneae. Our genome assembly expands the scope of sequenced genomes within the Cyperaceae family, encompassing the fifth genus. It also provides research resources on Cyperid evolution and wetland conservation. [ABSTRACT FROM AUTHOR]

Simple Summary: In the species-diverse butterfly genus Polyommatus, speciation is often driven by rapid changes in chromosome number and structure, resulting in cryptic species. These cryptic species are morphologically similar, but usually can be recognized relatively easily using chromosomal markers. In this work, we show that similar chromosome numbers can independently evolve in different phylogenetic lineages, resulting in species that are difficult to distinguish using routine cytogenetic techniques. We also demonstrate that the combined analysis of chromosomes and mitochondrial DNA barcodes is a simple and effective tool for identifying such cryptic species. The detection of cryptic species in complexes that have undergone recent speciation is often difficult, since many standard nuclear markers have not yet accumulated differences between closely related taxa, and differences in mitochondrial markers can be leveled out due to mitochondrial introgressions. In these cases, the use of derived chromosomal characters such as non-ancestral chromosomal numbers and/or unusual karyotype features may be a solution to the species delimitation problem. However, non-ancestral but similar karyotypes may arise secondarily as a result of homoplastic evolution, and their interpretation as homologies may lead to incorrect taxonomic conclusions. In our study, we show that the combined use of mitochondrial DNA barcodes and karyotypes helps to solve this problem and identifies cryptic species in situations where each of these markers does not work individually. Using this approach, we show that the fauna of Armenia and adjacent Iran includes the following cryptic taxa of the Polyommatus ripartii species complex (haploid chromosome number, n in parentheses): P. ripartii paralcestis (n = 90), P. ripartii kalashiani, subsp. nov (n close to 90), P. emmeli, sp. nov. (n = 77–79), P. keleybaricus, sp. nov. (n = 86), P. demavendi belovi (n = 73–75), P. demavendi antonius, subsp. nov. (n = 71–73), P. admetus anatoliensis (n = 79) and P. eriwanensis (n = 29–34). Polyommatus admetus yeranyani is synonymized with P. admetus anatoliensis. [ABSTRACT FROM AUTHOR]

Chromosomal rearrangements can interfere with the disjunction and segregation of other chromosome pairs not involved in the rearrangements, promoting the occurrence of numerical abnormalities in resulting gametes and predisposition to trisomy in offspring. This phenomenon of interference is known as the interchromosomal effect (ICE). Here we report a prenatal case potentially generated by ICE. The first-trimester screening ultrasound of the pregnant woman was normal, but the NIPT indicated a high risk for three copies of chromosome 21, thus suspecting trisomy 21 (T21). After a comprehensive clinical evaluation and genetic counseling, the couple decided to undergo amniocentesis. The prenatal karyotype confirmed T21 but also showed a balanced translocation between the long arm of chromosome 15 (q22) and the long arm of chromosome 22. The parents' karyotypes also showed that the mother had the 15;22 translocation. We reviewed T21 screening methods, and we performed a literature review on ICE, a generally overlooked phenomenon. We observed that ours is the first report of a prenatal case potentially due to ICE derived from the mother. The recurrence risk of aneuploidy in the offspring of translocated individuals is likely slightly increased, but it is not possible to estimate to what extent. In addition to supporting observations, there are still open questions such as, how frequent is ICE? How much is the aneuploidy risk altered by ICE? [ABSTRACT FROM AUTHOR]

Purpose: From a diagnostic standpoint, certain approaches to genetic screening in clinical practice remain ambiguous in the era of assisted reproduction. Even the most current guidelines do not provide definite guidance on testing protocols, leaving clinicians to carefully determine which tests best serve patients struggling with infertility. The lack of uniformity in the current practice of male fertility evaluation can prove to be quite costly, thus necessitating healthcare practitioners to carefully appraise the necessity and weigh the advantages against potential economic and psychological detriments. The objective of this review is to map the existing literature on the general topic of the clinical indications of routine karyotyping and/or AZF screening in infertile men, identify key concepts, determine where the gaps are, and lastly, provide an overview of the conclusions drawn from a body of knowledge that varies widely in terms of methodologies or disciplines. Materials and Methods: A thorough search was conducted for the published findings up until July 2023, utilizing PubMed (MEDLINE). This comprehensive search involved the use of specific search keywords, either individually or in combination. The search terms employed were as follows: “Karyotype”, "Klinefelter" or "KS" or "47,XXY", "AZF" or "Azoospermi*" and/or "microdeletion*" in the title or abstract. Once the titles and abstracts of selected articles were obtained, the complete texts of linked papers were meticulously scrutinized. Results: A total of 191 records were identified from PubMed. During screening, 161 records (84.3%) were eliminated. Finally, 30 papers were included in this scoping review, which was conducted in 18 countries. The number of sequence tag sites (STSs) used in the studies varied from 5 to 59. The rate of AZF deletions among patients with NOA ranged from 1.3% to 53%. The mean frequency was estimated to be 5.6%. The rate of YCM among patients with XXY karyotype was nil in 19 out of 30 studies (63%), whilst, in the remaining studies, the rate varied from 0.8% to 67%. Conclusion: This review provides insights into managing male infertility. The presence of spermatozoa in ejaculation and successful surgical retrieval cannot be excluded for individuals with AZFb/AZFbc microdeletions. Screening for Y chromosome microdeletions is not needed for mosaic or classic KS. Only 1% of individuals with sperm concentration exceeding 1×106 sperm/mL and less than 5×106 sperm/mL exhibit AZF microdeletions; therefore, testing referral for such populations may need reassessment. Individuals with mosaic monosomy X karyotype and certain chromosomal anomalies should be referred for AZF deletion screening. These findings have implications for male infertility management and future research. [ABSTRACT FROM AUTHOR]

The widespread Palearctic rodent species gray hamster Nothocricetulus migratorius has a karyotype with a stable number of chromosomes 2n = 22 throughout the entire range of its habitat. We found gray hamsters with diploid number of chromosomes 2n = 24 locally distributed in the Qurama Ridge of the Tian Shan. A new karyotype and analysis of G- and NORs-bands of differentially stained chromosome sets were described for the first time. The described karyotype differs from the 22-chromosomal karyotype of gray hamsters by the Y-chromosome morphology and the presence of an additional pair of heteromorphic small chromosomes. Molecular genetic analysis revealed genetic divergence of 24- and 22-chromosomal forms of N. migratorius, and the differences between them in mitochondrial markers are comparable and in terms of nuclear markers exceed the differences between C. barabensis (2n = 20) and C. psevdogriseus (2n = 24). The data obtained give grounds to discuss the taxonomic status of the 24-chromosomal form of gray hamsters from the Qurama Ridge and consider the differentiation of N. migratorius karyomorphs as a stage of chromosomal speciation. [ABSTRACT FROM AUTHOR]

Background: The clinical features of Turner syndrome (TS) involve multiple organ system dysplasia, among which growth retardation and gonadal dysplasia are the most important clinical phenotypes. Methods: G banding karyotype analysis, chromosome microarray (CMA), and fluorescence in situ hybridization (FISH) were used for prenatal diagnosis of fetal chromosomes. Results: The result of fetal chromosome karyotype analysis was 46,XX. CMA showed arr[GRCh38]Xp22.33 p22.13(251888_18176046)x1,Xq27.1q28(140998347_156003433)x3. FISH indicated that the short arm end fragment of X chromosome was monomer and the long arm end fragment was trisomy. Conclusions: The fetal chromosome karyotype was normal, but CMA indicated that there was deletion and duplication of X chromosome. FISH verified the CMA results, locating the deletion and duplication fragments. CMA and FISH make up for the shortcomings of chromosome karyotype analysis technique. It is suggested that multiple detection methods should be applied in genetic prenatal diagnosis [ABSTRACT FROM AUTHOR]

Inherited disorders are one of the reasons of infertility and economic losses for the equine industry. The detection rate of chromosomal abnormalities is rising due to the use of sex chromosome linked molecular markers. Here, a rare sterile five-year-old Arabian mare with XY male chromosomes is presented. The phenotype was corresponded to female with normal external genitalia. By transrectal ultrasound, there were hypoplastic ovaries and aplasia of uterine horns. Chromosome analysis was accomplished on blood samples using conventional, and G-banding techniques and confirmed by PCR and hormonal analysis. Although the karyotype (2n = 64, XY) revealed a typical male arrangement, it was SRY gene negative and amelogenin gene positive for X and Y chromosome. Hormonal analysis showed slight estrogenic activity of the ovary, but low progesterone and anti-mullerian hormones levels. In conclusion, this case of sex reversal mare (SRY-negative 64, XY) was recorded using cytogenetic, genetic, and hormonal analysis. Cytogenetics and molecular screening could be used as a fast approach for reproductive disorders evaluation in equine to save money, effort, and time of breeders. [ABSTRACT FROM AUTHOR]

Short-read genome sequencing (GS) holds the promise of becoming the primary diagnostic approach for the assessment of autism spectrum disorder (ASD) and fetal structural anomalies (FSAs). However, few studies have comprehensively evaluated its performance against current standard-of-care diagnostic tests: karyotype, chromosomal microarray (CMA), and exome sequencing (ES). To assess the clinical utility of GS, we compared its diagnostic yield against these three tests in 1,612 quartet families including an individual with ASD and in 295 prenatal families. Our GS analytic framework identified a diagnostic variant in 7.8% of ASD probands, almost 2-fold more than CMA (4.3%) and 3-fold more than ES (2.7%). However, when we systematically captured copy-number variants (CNVs) from the exome data, the diagnostic yield of ES (7.4%) was brought much closer to, but did not surpass, GS. Similarly, we estimated that GS could achieve an overall diagnostic yield of 46.1% in unselected FSAs, representing a 17.2% increased yield over karyotype, 14.1% over CMA, and 4.1% over ES with CNV calling or 36.1% increase without CNV discovery. Overall, GS provided an added diagnostic yield of 0.4% and 0.8% beyond the combination of all three standard-of-care tests in ASD and FSAs, respectively. This corresponded to nine GS unique diagnostic variants, including sequence variants in exons not captured by ES, structural variants (SVs) inaccessible to existing standard-of-care tests, and SVs where the resolution of GS changed variant classification. Overall, this large-scale evaluation demonstrated that GS significantly outperforms each individual standard-of-care test while also outperforming the combination of all three tests, thus warranting consideration as the first-tier diagnostic approach for the assessment of ASD and FSAs.

BackgroundPatients with acute myeloid leukemia (AML) who have tumor protein p53 (TP53) mutations or a complex karyotype have a poor prognosis, and hypomethylating agents are often used. The authors evaluated the efficacy of entospletinib, an oral inhibitor of spleen tyrosine kinase, combined with decitabine in this patient population.MethodsThis was a multicenter, open-label, phase 2 substudy of the Beat AML Master Trial (ClinicalTrials.gov identifier NCT03013998) using a Simon two-stage design. Eligible patients aged 60 years or older who had newly diagnosed AML with mutations in TP53 with or without a complex karyotype (cohort A; n = 45) or had a complex karyotype without TP53 mutation (cohort B; n = 13) received entospletinib 400 mg twice daily with decitabine 20 mg/m2 on days 1-10 every 28 days for up to three induction cycles, followed by up to 11 consolidation cycles, in which decitabine was reduced to days 1-5. Entospletinib maintenance was given for up to 2 years. The primary end point was complete remission (CR) and CR with hematologic improvement by up to six cycles of therapy.ResultsThe composite CR rates for cohorts A and B were 13.3% (95% confidence interval, 5.1%-26.8%) and 30.8% (95% confidence interval, 9.1%-61.4%), respectively. The median duration of response was 7.6 and 8.2 months, respectively, and the median overall survival was 6.5 and 11.5 months, respectively. The study was stopped because the futility boundary was crossed in both cohorts.ConclusionsThe combination of entospletinib and decitabine demonstrated activity and was acceptably tolerated in this patient population; however, the CR rates were low, and overall survival was short. Novel treatment strategies for older patients with TP53 mutations and complex karyotype remain an urgent need.

Disorders of sexual development (DSD) are associated with atypical chromosomal, gonadal, or phenotypic sex. It is likely that the number of cases of DSD are underestimated in the equine population. Monorchidism in the horse is very rare. This case report describes the clinical assessment of a phenotypic mare with stallion-like behavior which led to the diagnosis of a DSD. A 4-year-old Quarter Horse mare presented in good body condition, with normal external genitalia for a mare, and normal mammary glands with two bilaterally symmetric teats. No uterus, cervix, or gonads were detected on transrectal palpation. Transrectal ultrasonography revealed a single gonad in the right dorsal abdomen with the morphologic appearance of a testicle. Presurgical hormonal evaluation revealed elevated serum testosterone and anti-Müllerian hormone (AMH) concentrations. The right gonad was successfully removed via standing exploratory laparoscopy and submitted for histopathology. No gonad was identified on the left side during laparoscopy. Histopathologic examination confirmed that the excised gonad was a testicle. Cytogenetic and molecular analysis revealed a 64,XY, SRY-positive chromosomal constitution. Hormonal evaluation 5 weeks after surgery revealed low serum testosterone and AMH levels. A diagnosis of monorchidism was based on ultrasound examination, laparoscopic exploration of the abdomen, removal of a single gonad, and a subsequent decrease in serum testosterone and AMH concentrations to basal levels. In summary, a combination of clinical signs, endocrine evaluation, chromosomal and molecular analysis, and histopathology can be used in the diagnosis of DSD conditions.