Genetics Laboratory Research Articles

High definition 3D models of small legume seed: a close up application to support ancient plant genomics

Abstract. Legume seeds are essential for nutrition and play crucial roles in food security, climate change mitigation, biodiversity conservation, and sustainability. Among them, the common bean (Phaseolus vulgaris L.) is the most widely cultivated and vital to the food value chain. Originating in Mesoamerica and independently domesticated there and in the Andes about 8,000 years ago, common beans have adapted to diverse environments globally following the Columbian exchange. Ancient DNA (aDNA) sequencing offers insights into the bean’s adaptation and evolution, but traditional methods struggle with detailed 3D phenotypic analysis due to the beans’ small size and the destructive nature of aDNA extraction. Expensive high-detail optical instruments are not commonly available in genetic labs. This research proposes macro-photogrammetry as a low-cost technique for creating detailed 3D digital replicas of beans. By using samples from archaeological sites in northern Peru, this method preserves phenotypic information despite the destruction of physical samples during DNA extraction. The study details the process of capturing high-detail images with specialized equipment to build a digital phenotype library, preserving the morphological features of ancient beans for further study and future comparison with modern varieties.

The role and prognostic value of the interleukin 2 gene at multiple myeloma

Aim of the study. To determine the relationship of the polymorphic locus of the IL2 genetic marker (T-330G) with the risk of complicated course of multiple myeloma.Materials and methods. The molecular analysis of the IL2 (T-330G) gene was carried out in the laboratory of Molecular Genetics, cytogenetics and FISH at the Republican Specialized Scientific and Practical Medical Center of Hematology of the Republic of Uzbekistan (Tashkent city). 101 patients aged 34 to 72 years with a reliably established diagnosis of multiple myeloma were examined. IL2 polymorphism (T-330G) was genotyped by real-time PCR using primers from Litech (Russia) using an Applied Biosystems thermal cycler (USA).Results and discussion. An increased risk of complicated course of multiple myeloma compared with healthy individuals was found to be 1.6 times higher among carriers of the mutant G allele (χ2=4.3; p=0.05). Statistically significant differences between unfavorable loci in the group of patients with combined complications of multiple myeloma compared with the control were revealed.Conclusions. The risk of developing multiple myeloma compared with healthy individuals is 1.6 times higher among carriers of the mutant G allele (χ2=4.3; p=0.05) according to the polymorphism of the IL2 gene (T-330G). Along with this, a statistically significant relationship was established between unfavorable loci (G and G/G) with an increase of 2.5 (χ2=9.4; p=0.01) and 3.5 times (χ2=5.3; p=0.03) the risk of developing multiple myeloma complicated by plasmacytoma in combination with nephropathy.

A success story in a low-resource setting: Advancing cancer diagnostics in underserved region

Abstract Introduction/Objective Shefaa Al Orman Hospital (SOH) is the first oncology hospital in Luxor. Since its inception in 2016, the hospital has served approximately 45,000 patients from Luxor and other governorates. The hospital’s lab provides excellent routine laboratory services. However, there is a pressing demand for molecular genetics and cytogenetics, that is necessary for diagnosis of hematological malignancies. This strategic expansion faced several challenges: lack of expertise and the need for special expensive equipment and reagents. Methods/Case Report The project started with collaboration with oncologists to convene on a simplified panel with the aim of prioritizing clinically relevant tests. At first, these investigations were referred to a specialized oncology hospital in Cairo. This entailed 2 problems: transport of samples led to deterioration of the quality of some samples and delay in results reporting. In the second phase, experienced consultants from Cairo were enlisted to provide training and mentorship to local staff. Infrastructure setup included the allocation of dedicated laboratory space and the procurement of specialized equipment. Test method selection was used to achieve successful transition. In cytogenetics, we started with FISH technique which, while more expensive, gave rapid and reliable results. After gaining experience, the technique of conventional karyotyping was established. On the other hand, in molecular genetics, we resorted to custom made reagent mix preparation, instead of commercial kits. This approach, while more laborious, is less expensive. Results (if a Case Study enter NA) With the gradual successful in-house setup of these tests, there has been a notable reduction of out-sourced samples to 5%. This has led to shorter result turnaround times. Cytogenetics and FISH improved from 4 to 1 week, while molecular genetics decreased from 4 to 2 weeks. Moreover, the local processing of samples has minimized the risk of sample damage during transportation, ensuring the integrity of test results. Conclusion The establishment of the Molecular Genetics and Cytogenetics Laboratory at SOH represents a significant milestone. The hospital has successfully overcome logistical challenges and enhanced diagnostic capabilities for hematological malignancies, contributing to better patient outcomes and satisfaction. Moving forward, the continued investment in infrastructure, personnel training, and quality assurance measures will further strengthen the hospital’s capacity to provide high-quality cancer care to the community.

7704 Generating pituitary cells from iPSCs of patients with child onset congenital hypopituitarism harboring PROP1 pathogenic allelic variants

Abstract Disclosure: J.M. Marques: None. F. Fattahi: None. L.R. Carvalho: None. Introduction: Reduced or absent levels of pituitary hormones due to genetic or organic causes is known as combined pituitary hormone deficiency (CPHD) and PROP1 is the most frequent cause of CPHD, but more than 85% of the cases are without molecular diagnosis. Novel Techniques such as induced pluripotent stem cells (iPSC), can be used to generate patient’s specific pituitary cells for disease modeling. Aim: To generate iPSC from CO-CH patients’ peripheral blood mononuclear cells (PBMC) and differentiate them into pituitary cells. Methods: Peripheral blood was collected from 2 siblings (HC1 and HC2) harboring compound heterozygous PROP1 allelic variants and their parents (HC3 and HC4). PBMCs were isolated and expanded with MNC medium prior to nucleoporation and transfection with plasmids containing pluripotency genes (OCT4 and SOX2; cMYC and KLF). After 48h, transfected cells were transferred to Geltrex coated plates and fed with E8, NaB and βFGF until pluripotent colonies were formed. iPSCs were differentiated into cranial placode cells using pituitary placode conditioned medium (PPCM) until day 15. For pituitary cell maturation, pituitary conditioned medium (PPCM without SB431542) was used until day 30. qPCR and flow cytometry using the Kit BD Stemflow - Human Pluripotent Stem Cell Transcription Factor Analysis (BD Biosciences) for stem cells and the eBioscience™ Foxp3 / Transcription Factor Fixation/Permeabilization (Thermo Fisher Scientific) for the placode and pituitary markers were used. Pituitary hormonal levels from cell supernatant were measured at the Laboratory of Hormones and Molecular Genetics - HCFMUSP. Results: we generated iPSCs from patient and control blood cells. iPSCs were more than 90% positive for the pluripotency markers SOX2, NANOG, OCT3/4. Pituitary cells were generated with positivity for the placode marker SIX1 (58%) and pituitary markers, as LHX3 (56%) and PROP1 (59%), at day 15. Control cell lineages expressed higher levels of pituitary hormones at days 15 and 30 at RNA and protein levels while patient cells failed to induce hormone-producing cells properly. Pituitary hormonal release was impaired in patients pituitary cells. Conclusion: CO-CH patients’ PBMC were dedifferentiated to a pluripotent state, being able to generate pituitary hormone producing cells. Patient cells were able to mimic human phenotype. These cells can be used, in the future, to study CO-CH basis in patients specific background, to develop pituitary disease modeling as well as test possible new treatments. Presentation: 6/3/2024

8097 A Rare Case of a Renal Cell Carcinoma and Pheochromocytoma Collision Tumor

Abstract Disclosure: R. Daas: None. S. Logan: None. W. Sexton: None. V. Chumbalkar: None. M. McGettigan: None. C. Veloski: None. Background: Collision tumors are neoplasms comprised of two or more distinct tumors. A collision tumor consisting of a renal cell carcinoma and pheochromocytoma is extremely rare with less than five cases reported in the literature. Clinical Case: A 64-year-old male with a history of hypertension and type 2 diabetes presented with hematuria. The patient was on 4 anti-hypertensive medications at the time. He noted increased road rage, anxiety, tremors, and insomnia. A CT abdomen/pelvis revealed an exophytic 8.6 x 7.9 cm left upper pole renal mass and a 7.5 x 6.2 cm left adrenal mass. Contrast-enhanced MRI abdomen showed a left upper/interpolar renal mass with heterogenous peripheral enhancement, restricted diffusion, and areas of central necrosis along with a heterogenous, enhancing left adrenal mass with associated restricted diffusion and areas of cystic necrosis. Both masses had associated areas of hyperintense signal on T1 fat-suppressed MRI suggestive of hemorrhage. Lab work revealed ACTH 26.6 pg/mL (7.2-63.3), cortisol 13 ug/dL (3.7-19.4), aldosterone 23.1 ng/dL (4-31), renin 4 ng/mL/hr (0.5-4), dopamine 136 pmol/L (<240), epinephrine 121 pmol/L (<330), norepinephrine 23,666 pmol/L (1050-4800), plasma normetanephrines 12.3 nmol/L (0-0.89), plasma metanephrines <0.10 nmol/L (0-0.49), 24-hour urine normetanephrines 4,178 mcg (122-676) and 24-hour urine metanephrines 4,233mcg (224-832). The patient was treated with alpha blockade prior to left radical nephrectomy, left adrenalectomy and infrahilar paraaortic lymph node dissection. Despite aggressive preoperative alpha blockade with maximal doses of doxazosin, addition of metyrosine and other anti-hypertensives, he developed acute hypertension with adrenal mass manipulation during the procedure and then hypotension following removal. He was admitted to the ICU for management of hypotension with pressors and fluid resuscitation and did well. Post-operative labs showed plasma normetanephrines 0.98 nmol/L (0.00-0.89) and plasma metanephrines 0.11 nmol/L (0.00-0.49). Pathology revealed clear cell renal cell carcinoma (RCC) of the kidney. The adrenal gland showed 2 tumors; one consistent with pheochromocytoma and another intimately associated with the former consistent with metastatic RCC, morphology diagnostic of a collision tumor. Following surgical resection, the patient was referred for genetic testing, surveillance imaging and lab testing, and treated with pembrolizumab adjuvant therapy for stage IV RCC. Conclusion: Although collision tumors are rare, biochemical testing to rule out a pheochromocytoma must be performed for all indeterminate adrenal masses and presumed adrenal metastases of active malignancies. Presentation: 6/2/2024

Canadian College of Medical Geneticists: clinical practice advisory document – responsibility to recontact for reinterpretation of clinical genetic testing

BackgroundAdvances in technology and knowledge have facilitated both an increase in the number of patient variants reported and variants reclassified. While there is currently no duty to recontact for reclassified...

Genetic variant profiling of neonatal diabetes mellitus in Iranian patients: Unveiling 58 distinct variants in 14 genes.

Neonatal diabetes mellitus (NDM) is a rare non-immunological monogenic disorder characterized by hyperglycemic conditions primarily occurring within the first 6 months of life. The majority of cases are attributed to pathogenic variants in genes affecting beta-cell survival, insulin regulation, and secretion. This study aims to investigate the genetic landscape of NDM in Iran. We recruited a total of 135 patients who were initially diagnosed with diabetes at <12 months of age in Iran and referred to pediatric endocrinology clinics across the country. These patients underwent genetic diagnostic tests conducted by the Exeter Molecular Genetics Laboratory in the UK. The pathogenic variants identified were sorted and described based on type, pathogenicity (according to ACMG/AMP criteria), novelty, and the affected protein domain. Genetic defects were identified in 93 probands, presenting various pathogenic abnormalities associated with NDM and its associated syndromes. 76% of the patients were born as a result of consanguineous marriage, and a familial history of diabetes was found in 43% of the cases. A total of 58 distinct variants in 14 different genes were discovered, including 20 variants reported for the first time. Causative variants were most frequently identified in EIF2AK3, KCNJ11, and ABCC8, respectively. Notably, EIF2AK3 and ABCC8 exhibited the highest number of novel variants. These findings provide valuable insights into the genetic landscape of NDM in the Iranian population and contribute to the knowledge of novel pathogenic variants within known causative genes.

Prevalence of Human Papillomavirus (HPV) Infection and its Association With Pap Smear Findings Among Women Attending a Gynecology Clinic in Khorasan Razavi-Iran.

Human papillomavirus (HPV) infection is the most common sexual transmitted disease and Pap smears and HPV testing are crucial for early detection. Advancements in HPV testing improve accuracy, but optimal screening strategies are still debated. This cross-sectional study explores HPV genotypes and predictors of infection among Iranian women undergoing gynecological screening. Study Design: A retrospective cross-sectional study. Women undergoing their initial cervical screening enrolled in this study. Cervical cytology samples for Pap smear analysis were collected from women referred to the genetic laboratory of Academic Center for Education Culture and Research (ACECR), Khorasan Razavi, during gynecological visits, adhering to standardized liquid-based cytology protocols. These samples were obtained over a one-year period since January 2023. Statistical analyses were conducted using SPSS version 21.0, with a significance level set at P<0.05. A total of 328 women enrolled in this study. The mean age of participants was 36±11 years and the overall HPV prevalence was 37.5% (n=123). Among HPV-positive women, nearly half (48.7%) had a single HPV genotype. Genotypes 6 (13%), 16 (12.3%), and 53 (6.7%) were the most prevalent types. Notably, high-risk HPV genotypes (16 and 18 among all) were identified in one-fourth of the study population. Women with endocervical/transformation zone cells had 25% higher odds of having HPV infection, and having mild, moderate, and severe inflammation increased the odds of having HPV infection by 14%, 11%, and 20%, respectively. The considerably high prevalence of HPV infection highlights the significance of HPV prevention programs in this population. Neither bacterial vaginosis nor candida infection showed a direct link to HPV positivity.

The management of Familial Pulmonary Fibrosis in different medical settings: Where does that leave us? An Italian nationwide survey.

Familial Pulmonary Fibrosis (FPF) is an emerging group of interstitial lung diseases (ILDs) caused by mutations mainly involving "telomere-related genes" and "surfactant-related genes". Although, in 2023, European Respiratory Society proposed a statement for FPFs management, these still remain a burden. Our work aimed to evaluate the management and impact of FPF in three Italian different medical settings: University Hospitals (UHs), non-University Hospitals (N-UHs) and outpatient clinics. This survey was created by ILDs Study Group Società Italiana di Pneumologia/ Italian Respiratory Society (SIP-IRS) and diffused via email to all SIP-IRS members. The descriptive statistical analysis was conducted through GraphPad Prism software (version 8.0). Results: Twenty participants replied to the survey, of which 65% (13/20) worked at UH while the remaining 25% (6/20) and 5% (1/20) worked at N-UH and outpatient clinics, respectively. Centers with, at least, 150 ILD patients visits/year followed a higher number of FPF patients, regardless of University affiliation (p=0.0046). Despite significant discrepancies in genetic testing and availability of counselling were registered, no statistically significant differences in patients' anamnesis assessment were observed between UHs and N-UHs (p=0.4192 and p=0.6525). However, there were relevant differences in the number of FPF patients undergoing genetic assessment in the Centers with Genetics Lab or Unit inside the Hospital (p=0.0253). There was no consensus regarding the impact of FPF diagnosis on lung transplantation and screening of asymptomatic relatives. Similarly, no differences were reported in antifibrotic prescriptions between UHs and N-UHs. Although the typical UIP pattern was the most common radiological pattern observed in FPF patients, there were no differences in the prevalence of histopathological patterns between UH and N-UH. Improving pulmonologists' knowledge of the approach, diagnosis and management of FPF is a global medical topic. Scientific societies can provide significant support in raising physicians' awareness of this issue.

The current landscape of clinical exome and genome reanalysis in the U.S.

The majority of patients undergoing exome or genome sequencing receive a nondiagnostic result. Periodic reanalysis is known to increase diagnostic yield from exome sequencing, yet laboratory reanalysis practices are obscure. We sought to define the landscape of exome and genome reanalysis across clinical laboratories. Genetic testing registries were queried to identify eligible clinical genetic laboratories offering exome and/or genome sequencing in the United States. A survey administered to lab representatives investigated reanalysis offerings, policies, perceived uptake, bioinformatic steps, and billing options. The analysis consisted of descriptive statistics. Survey data were collected from 30 of 32 eligible laboratories (93%), comprising 28 exome products and 13 genome products. Reanalysis was widely available for both exomes (n = 27/28, 96%) and genomes (n = 12/13, 92%). Most participating laboratories required ordering providers to initiate reanalysis (n = 24/28, 86%). Most respondents estimated providers initiated reanalysis in less than 10% of all exomes (n = 12/22) or genomes (n = 6/9) sequenced. The approach to reanalysis varied greatly by laboratory. Laboratory approaches to exome and genome reanalysis are highly variable and typically require provider initiation. This could contribute to low reanalysis uptake and increased administrative burden on providers. Further work should emphasize development of clinical exome and genome reanalysis standards.

Biosynthesis and Characterization of Zinc Nanoparticles Using Strains of Pseudomonas and Actinobacteria

Research was carried out at the Green Nanotechnology Laboratory, University of Agricultural Sciences, Dharwad, Karnataka, with a specific emphasis on the biosynthesis of zinc nanoparticles using Pseudomonas and Actinobacteria. Collection and screening of Pseudomonas and Actinobacterial isolates were done at Microbial Genetics Laboratory, Department of Agricultural Microbiology, UAS, Dharwad. The ZnNPswere biosynthesized and characterized through UV-Visible spectroscopy, Particle Size Analyzer (PSA), Scanning Electron Microscope (SEM), Energy-dispersive X-ray spectroscopy (EDS), X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). The ZnNPs synthesized through Pseudomonas(AUDP209) and Actinobacteria(AUDT636)isolates exhibited peak values at 365 and 374 nm, with average sizes of 48.7 nm and 54.9 nm, respectively.SEM images revealed the spherical shape of the biosynthesized ZnNPs, while EDX confirmed the zinc ions presence in the biosynthesized samples and the crystalline structure of the ZnNPswas confirmed by XRD.FTIR spectroscopy confirmed the organic compounds present in the microbial extracts responsible for the capping and stabilizing agents for biosynthesis of ZnNPs.

Expanding the Russian allele frequency reference via cross-laboratory data integration: insights from 7452 exome samples.

Population allele frequency is crucially important for accurate interpretation of known and novel variants in medical genetics. Recently, several large allele frequency databases, such as the Genome Aggregation Database (gnomAD), have been created to serve as a global reference for such studies. However, frequencies of many rare alleles vary dramatically between populations, and population-specific allele frequency is often more informative than the global one. Many countries and regions, including Russia, remain poorly studied from the genetic perspective. Here, we report the first successful attempt to integrate genetic information between major medical genetic laboratories in Russia. We construct RUSeq, an open, large-scale reference set of genetic variants by analyzing 7452 exome samples collected in two major Russian cities-Moscow and St. Petersburg. An ∼10-fold increase in sample size compared to previous studies allowed us to characterize extensive genetic diversity within the admixed Russian population with contributions from several major ancestral groups. We highlight 51 known pathogenic variants that are overrepresented in Russia compared to other European countries. We also identify several dozen high-impact variants that are present in healthy donors despite being annotated as pathogenic in ClinVar and falling within genes associated with autosomal dominant disorders. The constructed database of genetic variant frequencies in Russia has been made available to the medical genetics community through a variant browser available at http://ruseq.ru.

Age estimation of burnt human remains through DNA methylation analysis.

The identification of human fire victims is a challenging task in forensic medicine. The heat-induced alterations of biological tissues can make the conventional anthropological analyses difficult. Even if the DNA profile of the victim is achieved, it is possible that no match can be found in a forensic DNA database, thus hindering positive identification. In such cases, any information useful to nail down a possible identity should be collected, such as DNA methylation analysis which could provide useful investigative leads. In the present study, five age-related epigenetic markers (ELOVL2, FHL2, KLF14, C1orf132, and TRIM59) were initially analysed in blood samples of 72 living Italian individuals of known age, using a Single Base Extension (SBE) assay. An age prediction model was built by multiple linear regression including all the markers (Mean Absolute Error, MAE: 3.15 years). This model was tested on 29 blood samples collected during autopsies from burnt human remains, already identified through DNA analysis, providing a MAE of 6.92 years. The model allowed a correct prediction in 79.3% of the cases (95% prediction interval), while six cases were associated with inaccurate predictions (min-max prediction error: 9.8-37.3 years). Among the different sample variables considered to explain these results, only the DNA degradation index was a relevant factor affecting the reliability of the predictions. In conclusion, the SBE typing of blood from burnt remains proved to be a reliable tool to estimate chronological age of most of the samples, also in consideration of its cost-effectiveness and the availability of CE sequencers in every forensic genetics laboratory.

Acute Myelomonocytic Leukemia Eosinophilic Variant: Epidemiological, Clinical, and Biological Profile at the Hassan II University Hospital of Fez

Introduction: Acute myelomonocytic leukemia with an eosinophilic component (AML4Eo) is a particular and rare hematological entity, representing between 5% and 8% of all acute myeloid leukemias. The aim of our work is to highlight the clinical and cytogenetic epidemiological particularities of this pathology at the Hassan II University Hospital in FES. Patients and Methods: Retrospective descriptive study spread over 14 years (January 2009-January 2023) including all patients diagnosed with LAM4Eo.Myelogram reading as well as immunophenotyping by flow cytometry on Cytomic FC500 were performed at the central hematology laboratory and cytogenetics at the genetics laboratory CHU HASSAN II of FES. Clinical data were collected from patients' medical charts. Results: Thirteen patients were enrolled. The mean age at diagnosis was 20.4 years, with a predominance of males (54%) most patients presented with an altered general condition, signs of cytopenia and a tumor syndrome. All patients presented with profound anemia associated with thrombocytopenia and hyperleukocytosis. Blood smears showed the presence of peripheral blasts in 80% of cases. All medullograms met the morphological criteria established by the FAB (Franco-American-British) classification for the diagnosis of LAM4Eo.Cytochemical staining with myeloperoxidase was performed on all medullograms and was positive in all cases. Immunophenotyping was carried out in 56% of patients, showing the existence of very immature blasts expressing myeloblast and monoblast markers. Four of our patients underwent cytogenetic studies, with no translocation or inversion of chromosome 16. Three patients died immediately after diagnosis. The other patients were put on chemotherapy, two of whom died during treatment. Discussion/Conclusion: LAM4Eo is a rare hematological malignancy with a better prognosis than other myeloid leukemias. Despite the development of cytogenetics and molecular biology techniques, morphological examination ..

A genetic screen of transcription factors in the Drosophila melanogaster abdomen identifies novel pigmentation genes.

Gene regulatory networks specify the gene expression patterns needed for traits to develop. Differences in these networks can result in phenotypic differences between organisms. Although loss-of-function genetic screens can identify genes necessary for trait formation, gain-of-function screens can overcome genetic redundancy and identify loci whose expression is sufficient to alter trait formation. Here, we leveraged transgenic lines from the Transgenic RNAi Project at Harvard Medical School to perform both gain- and loss-of-function CRISPR/Cas9 screens for abdominal pigmentation phenotypes. We identified measurable effects on pigmentation patterns in the Drosophila melanogaster abdomen for 21 of 55 transcription factors in gain-of-function experiments and 7 of 16 tested by loss-of-function experiments. These included well-characterized pigmentation genes, such as bab1 and dsx, and transcription factors that had no known role in pigmentation, such as slp2. Finally, this screen was partially conducted by undergraduate students in a Genetics Laboratory course during the spring semesters of 2021 and 2022. We found this screen to be a successful model for student engagement in research in an undergraduate laboratory course that can be readily adapted to evaluate the effect of hundreds of genes on many different Drosophila traits, with minimal resources.

Exploring the human genomic landscape: patterns of common homozygosity regions in a large middle eastern cohort.

Regions of Homozygosity (ROH) typically reflect normal demographic history of a human population, but may also relate to cryptic consanguinity, and, additionally, have been associated with specific medical conditions. The objective of this study was to investigate the location, size, and prevalence of common ROH segments in a Middle Eastern cohort. This retrospective study included 13 483 samples collected from all Chromosomal Microarray analyses (CMA) performed using Single Nucleotide Polymorphism (SNP) arrays at the genetic clinical laboratory of Rabin Medical Center between 2017-2023 (primary data set). An additional replication cohort including 100 842 samples from another SNP array platform, obtained from Maccabi Health Organization, was analyzed. Common ROH locations were defined as those ROH locations involving 1% or more of the samples. A total of 66 710 ROH segments, involving 13 035 samples (96.7%) were identified in the primary data set. Of the 4069 cytogenetic ROH locations, 68 were identified as common. The prevalence of non-common ROH was relatively high in affected individuals, and for acrocentric chromosomes, chromosomes associated with common trisomies, and non-imprinted chromosomes. In addition, differences in common ROH locations were observed between the primary and the replication cohorts. Our findings highlight the need for population-specific guidelines in determining ROH reporting cutoffs, considering factors such as population-specific prevalence and testing platform differences. Future research with larger, varied cohorts is essential to advance understanding of ROH's associations with medical conditions and to improve clinical practices accordingly.

Clinical Characterization of a National Cohort of Patients With Germline WT1 Variants Including Late-Onset Phenotypes

Background and AimsWT1-disorder is a recently introduced term for phenotypes associated with germline WT1-variants, including glomerulopathy, urogenital anomalies and Wilms tumor. Previous studies showed a bias towards missense variants in the DNA-binding/Zinc-finger domain of WT1 (exon 8/9) and patients with early-onset glomerulopathy. Thorough genotype-phenotype correlations including follow-up data on late-onset glomerulopathy risk are lacking. To characterize the genotypic and phenotypic spectrum of WT1 disorder, we describe a national cohort of individuals with WT1-variants. MethodsWe requested clinical and genetic data of all patients with germline WT1-variants at all Dutch genetic laboratories. ResultsWe identified 43 patients with pathogenic WT1-variants (truncating n=19, missense n=13, splice-site n=7 and deletions n=4). Wilms tumor was the only clinical manifestation in 10 patients, of whom nine females. Wilms tumor occurred in 18/19 patients with truncating variants, in 4/4 patients with deletions and was more rare in patients with missense- or splice-site variants. All patients with missense and 6/7 with splice-site variants developed CKD, versus 5/19 patients with truncating variants (three in adulthood, with kidney failure at 24, 26 and 41 years) and 1/4 with a deletion. Urogenital malformations occurred predominantly in 46,XY individuals. ConclusionAmong patients with WT1-variants, a genotype-phenotype correlation was observed for Wilms tumor risk and age of CKD onset. Although childhood-onset CKD was more common in patients with missense variants in the DNA-binding/Zinc-finger domain, other patients may develop CKD and kidney failure later in life. Therefore, life-long surveillance of kidney function is recommended. Alertness of WT1-variants is especially important for girls with Wilms tumor who often miss additional phenotypes.

Challenging Misconceptions about Race in Undergraduate Genetics.

Racial biases, which harm marginalized and excluded communities, may be combatted by clarifying misconceptions about race during biology lessons. We developed a human genetics laboratory activity that challenges the misconception that race is biological (biological essentialism). We assessed the relationship between this activity and student outcomes using a survey of students' attitudes about biological essentialism and color-evasive ideology and a concept inventory about phylogeny and human diversity. Students in the human genetics laboratory activity showed a significant decrease in their acceptance of biological essentialism compared with a control group, but did not show changes in color-evasive ideology. Students in both groups exhibited increased knowledge in both areas of the concept inventory, but the gains were larger in the human genetics laboratory. In the second iteration of this activity, we found that only white students' decreases in biological essentialist beliefs were significant and the activity failed to decrease color-evasive ideologies for all students. Concept inventory gains were similar and significant for both white and non-white students in this iteration. Our findings underscore the effectiveness of addressing misconceptions about the biological origins of race and encourage more research on ways to effectively change damaging student attitudes about race in undergraduate genetics education.

Detection of yellow mosaic virus resistance in Soybean (Glycine max L.) genotypes for yield and related traits

Background Twenty soybean genotypes were screened for Soybean yellow mosaic virus (YMV) identification which were collected from the Genetics and plant breeding farm laboratory. Methods To evaluate genetic studies of the chosen materials, the use of a net house assay upon sap inoculation made it easier to determine the trend of the Area Under Disease Progress Curve (AUDPC) in a controlled environment. Younger leaves from these plants were gathered and used for molecular analysis. Results Six genotypes were identified with semi-resistant disease reactions. Yellow mosaic virus-resistant and susceptible soybean genotypes were distinguished using a soybean “Rsv1-h” gene-based primer pair. To discover the existence of the Rsv-1-h gene responsible for YMV resistance, a genetic diversity panel of 15 soybean genotypes was investigated with two simple sequence repeat (SSR) markers. The BARCSOYSSR_13_1115 and BARCSOYSSR_13_1173 primer sets were employed to validate the desired genes in these genotypes. Conclusion The genotypes HIHS-WIHS, MINA-HAI, Shohag, G-2120, BRAGG, and BS-3 presented the major gene Rsv1-h, which is said to be essential for yellow mosaic virus resistance and can provide better yield also than other genotypes. The six soybean genotypes with the Rsv1-h gene may be resistant to mosaic virus and could be exploited as a source of improved breeding material in the future.

ANALYSIS OF STAFFING OF THE MEDICAL AND GENETIC SERVICE IN UKRAINE

Aim. To study the staffing conditions of the medical-genetic service in Ukraine during the period from 2008 to 2022 and to forecast the staffing dynamics until 2026. Materials and Methods: Statistical data from healthcare institutions in Ukraine that provided medical-genetic services during the period from 2008 to 2022 were analyzed, based on reporting forms 21, 17, 49 from the City Health Center website http://medstat.gov.ua/ukr/statdan.html and http://medstat.gov.ua/ukr/statdanMMXIX.html. Single-factor linear regression models were applied for mathematical modeling, with quality assessment conducted using the coefficient of determination R². Calculations were performed using MedCalc® Statistical Software (v.22.009). Results: Compared to 2008, there was a significant reduction in the number of full-time positions of heads of offices (genetic doctors), genetic doctors, and genetic laboratory doctors in Ukraine in 2022 by -45.24%, -24.83%, and -23.39%, respectively. A significant decrease in the number of these positions was determined (p&lt;0.05). In 2022, compared to 2008, there was also a decrease in the staffing rate of all full-time positions with occupied positions of the personnel of centers/offices. Specifically, for genetic doctors, the staffing rate in 2022 was 69.62% compared to 85.37% in 2008, and for genetic laboratory doctors, it was 57.54% compared to 94.62% in 2022 and 2008, respectively. The level of provision of geneticists (individuals) in Ukraine in 2022 was 1.99 per 1 million population, which is -16.03% less than in 2008. Furthermore, compared to 2008, the share of vacant positions for genetic doctors in 2022 increased 2-fold, and for genetic laboratory doctors almost 8-fold. According to the results of mathematical forecasting, in 2026, a further decrease in the availability of genetic doctors to the level of 1.77 per 1 million population is predicted (R²=0.88, p&lt;0.001). Results. Compared to 2008, there was a significant reduction in the number of full-time positions of heads of offices (genetic doctors), genetic doctors, and genetic laboratory doctors in Ukraine in 2022 by -45.24%, -24.83%, and -23.39%, respectively. A significant decrease in the number of these positions was determined (p&lt;0.05). In 2022, compared to 2008, there was also a decrease in the staffing rate of all full-time positions with occupied positions of the personnel of centers/offices. Specifically, for genetic doctors, the staffing rate in 2022 was 69.62% compared to 85.37% in 2008, and for genetic laboratory doctors, it was 57.54% compared to 94.62% in 2022 and 2008, respectively. Furthermore, in comparison with 2008, the share of vacant positions for genetic doctors in 2022 increased in 2-fold, and for genetic laboratory doctors almost in 8-fold. According to the results of mathematical forecasting, in 2026, a further decrease in the availability of genetic doctors to the level of 1.77 per 1 million population is predicted (R²=0.88, p&lt;0.001). Conclusions. Based on the analysis of staffing of the medical-genetic service in Ukraine during the period 2008-2022, conclusions can be drawn about the reduction in the number of full-time positions and the availability of genetic doctors in the medical-genetic service in Ukraine. To ensure proper medical-genetic counseling for the population, it is necessary to develop an effective strategy for improving the organization of the medical-genetic service, capable of providing comprehensive support for patients with genetic pathologies.