Miruna Gug

Background/Objectives: Prenatal testing strategies for suspected monogenic disorders have changed considerably over the past decades, alongside evolving referral indications and increasing availability of genomic technologies. Longitudinal, practice-based data describing these changes within routine clinical care remain limited, particularly in Central and Eastern Europe. We describe changes in referral indications, testing strategies, and molecular findings over 22 years in a single-center retrospective descriptive case series from Western Romania. Methods: We conducted a retrospective analysis of 52 pregnancies investigated for suspected monogenic disorders between 2004 and 2026. Forty pregnancies were evaluated through a diagnostic pathway, while a separate group of 12 pregnancies underwent cell-free DNA (cfDNA)-based monogenic screening; these were analyzed as distinct clinical pathways. For the diagnostic pathway, clinical indications, testing strategies, and molecular findings were analyzed across three retrospectively defined study periods (2004–2013, 2014–2019, and 2020–2026). Results: Within the diagnostic pathway, referral patterns shifted from predominantly family-history-based testing (55.6% of cases in 2004–2013) toward indications arising from positive parental carrier screening and fetal ultrasound abnormalities in later study periods. Testing strategies expanded from predominantly targeted single-gene testing and multiplex ligation-dependent probe amplification (MLPA) to include gene panels and whole-exome sequencing (WES). Among 34 pregnancies with fetal molecular evaluation, 7 (20.6%) had a confirmed disease-causing finding, 9 (26.5%) had carrier-only outcomes, 15 (44.1%) were classified as unaffected, and 3 (8.8%) had non-classic molecular findings. Incidental or additional molecular findings beyond the primary testing indication were identified in 6 of 34 pregnancies with fetal molecular evaluation (17.6%) and required case-specific interpretation and genetic counseling. The 12 cfDNA-based monogenic screening pregnancies constituted a separate, non-diagnostic screening pathway and were analyzed independently from the diagnostic pathway. Conclusions: Over 22 years, referral indications for suspected monogenic disorders broadened alongside an expansion of prenatal testing strategies from predominantly targeted familial testing to a wider range of genomic approaches. These findings describe temporal changes within a single-center clinical practice and should not be interpreted as evidence that changes in testing strategy improved diagnostic performance or pregnancy outcomes. Invasive diagnostic testing and cfDNA-based monogenic screening represent distinct clinical pathways and should be interpreted separately.

Background: Neurofibromatosis type 1 (NF1) is a multisystem genetic disorder characterized by marked phenotypic heterogeneity. This study aimed to describe the clinical spectrum and multisystem burden of NF1 in a regional retrospective cohort. Methods: We retrospectively analyzed 77 unique patients with NF1 after removal of duplicate records. Demographic characteristics, cutaneous manifestations, extracutaneous involvement, therapeutic interventions, and disease evolution were evaluated. An exploratory multisystem burden score was calculated using six extracutaneous clinical domains. Results: Café-au-lait macules were present in all patients, axillary/inguinal freckling in 93.5%, cutaneous neurofibromas in 63.6%, and plexiform tumors in 28.6%. Psychiatric/behavioral (67.5%), skeletal (57.1%), ophthalmological (51.9%), and neurological (45.5%) involvement were common. More than half of the cohort had involvement of at least three extracutaneous domains. Plexiform tumors were more common in patients with progressive or unfavorable disease compared to those with stationary disease. Progressive or unfavorable evolution was associated with a higher frequency of plexiform tumors. Therapeutic interventions were reported descriptively. Conclusions: NF1 showed substantial multisystem involvement beyond its characteristic cutaneous manifestations. Plexiform tumors were associated with a more complex clinical course. The proposed exploratory multisystem burden score may facilitate descriptive assessment of disease breadth but requires prospective validation.

Reproductive genetic carrier screening (RGCS) is recommended preconceptionally or early pregnancy to identify the risk of autosomal recessive (AR) disorders in healthy couples. Data on shared carrier status at the couple’s level remains limited in Eastern Europe. This study presents the first couple-based RGCS analysis in Western Romania. We retrospectively analyzed RGCS results from 247 couples with no known consanguinity at the time of evaluation (494 apparently unrelated individuals, aged 22–52 years), assessed at a single genetic center between 2020–2024. Next-generation sequencing was performed using an expanded panel targeting 302 genes, including 300 genes associated with AR onset disorders. This analysis was accompanied by both pre- and post-test genetic counseling. The prevalence of individual and shared carrier status and reproductive risk was assessed. Pathogenic or likely pathogenic (PLP) variants were identified in the study cohort, with an overall couple carrier frequency of 64.37%. Shared carrier status for PLP variants in the same gene was identified in 17.4%, including three couples carrying pathogenic variants in two shared genes. Additionally, 46.96% of couples carried pathogenic variants in different genes without overlapping. The most frequently shared genes with PLP variants were HFE, CFTR, SMN1, BTD, and COL7A1; 14 additional shared genes with PLP variants were associated with severe, early-onset disorders. Forty-three couples were deemed high risk for AR conditions. Their reproductive choices varied, including in vitro fertilization or proceeding with pregnancy with or without prenatal testing. Couple-based RGCS revealed a substantial burden of shared AR carrier status in Western Romania, supporting the implementation of population-level screening programs to improve reproductive risk assessment and informed decision-making.

Autosomal recessive (AR) disorders represent a significant public health challenge, as asymptomatic carriers are often unaware of their reproductive risks. This study provides the first comprehensive assessment of AR gene variant frequencies and their molecular landscape in a fertile Western Romanian population. Genetic results from 604 unrelated, unaffected Caucasian individuals of reproductive age, tested at a single genetic center between 2020 and 2024, were retrospectively analyzed. Next-generation sequencing (NGS) with a multi-gene panel targeting 300 AR-associated genes was used for molecular profiling. Variants were identified in 156 genes, with 75% of individuals carrying at least one AR variant (mean 1.77 variants/person). A subgroup with >3 pathogenic variants comprised 7.5%, posing a notable risk for future offspring. The most frequent variants were detected in HFE (1:5), CFTR (1:9), BTD (1:16), GJB2 (1:17), and CYP21A2 (1:19). Four variants (HFE, c.187C>G; BTD, c.1330G>C; CFTR, c.1210-34TG[11]T[5]; GALT, c.-119_-116del) were particularly prevalent, each exceeding 3% frequency. Considerable allelic heterogeneity was observed for distrinctive variants in CFTR (14), PAH (12), USH2A (12), and ATP7B (9). Several variants were linked to severe disorders, with CFTR, GALT, ATP7B, and SMN1 identified as “red zone” genes associated with high morbidity and mortality. Low-frequency variants formed a “long tail” (83.9%), reflecting marked population heterogeneity and potential hidden disease risks. The study reveals high allelic diversity and a strong prevalence of AR variants in Western Romania. Variant-based gene classification supports population-level screening, highlighting the public health value of a national program to identify carriers and prevent severe inherited disorders.

Background: Breast cancer remains the most prevalent malignancy in women worldwide, characterized by remarkable genetic, molecular, and clinical heterogeneity. Traditional preclinical models have significantly advanced our understanding of tumor biology, yet consistently fall short in recapitulating the complexity of the human tumor microenvironment (TME), immune, and metastatic behavior. In recent years, breast cancer-on-a-chip (BCOC) have emerged as powerful microengineered systems that integrate patient-derived cells, stromal and immune components, and physiological stimuli such as perfusion, hypoxia, and acidic milieu within controlled three-dimensional microenvironments. Aim: To comprehensively review the BCOC development and application, encompassing fabrication materials, biological modeling of key subtypes (DCIS, luminal A, triple-negative), dynamic tumor–stroma–immune crosstalk, and organotropic metastasis to bone, liver, brain, lungs, and lymph nodes. Methods: We selected papers from academic trusted databases (PubMed, Web of Science, Google Scholar) by using Breast Cancer, Microfluidic System, and Breast Cancer on a Chip as the main search terms. Results: We critically discuss and highlight how microfluidic systems replicate essential features of disease progression—such as epithelial-to-mesenchymal transition, vascular invasion, immune evasion, and therapy resistance—with unprecedented physiological relevance. Special attention has been paid to the integration of liquid biopsy technologies within microfluidic platforms for non-invasive, real-time analysis of circulating tumor cells, cell-free nucleic acids, and exosomes. Conclusions: In light of regulatory momentum toward reducing animal use in drug development, BCOC platforms stand at the forefront of a new era in precision oncology. By bridging biological fidelity with engineering innovation, these systems hold immense potential to transform cancer research, therapy screening, and personalized medicine.

Background: Understanding the interactions between age and comorbidities is crucial for assessing COVID-19 mortality, particularly in patients with cardiac and pulmonary conditions. This study investigates the relationship between comorbidities and mortality outcomes in a cohort of hospitalized COVID-19 patients, emphasizing the interplay of age, cardiac, and pulmonary conditions. Methods: We analyzed a cohort of 3005 patients hospitalized with COVID-19 between 2020 and 2022. Key variables included age, comorbidities (diabetes, cardiac, pulmonary, and neoplasms), and clinical outcomes. Chi-square tests and logistic regression models were used to assess the association between comorbidities and mortality. Stratified analyses by age, diabetes, and pulmonary conditions were conducted to explore interaction effects. Additionally, interaction terms were included in multivariable logistic regression models to evaluate the combined impact of age, comorbidities, and mortality. Results: Cardiac conditions such as hypertension, ischemic cardiopathy, and myocardial infarction showed significant protective effects against mortality in younger patients and in those without pulmonary conditions (p < 0.001). However, these protective effects were diminished in older patients and those with pulmonary comorbidities. Age was found to be a significant modifier of the relationship between cardiac conditions and mortality, with a stronger protective effect observed in patients under the median age (p < 0.001). Pulmonary comorbidities significantly increased the risk of mortality, particularly when co-occurring with cardiac conditions (p < 0.001). Diabetes did not significantly modify the relationship between cardiac conditions and mortality. Conclusions: The findings highlight the complex interactions between age, cardiac conditions, and pulmonary conditions in predicting COVID-19 mortality. Younger patients with cardiac comorbidities show a protective effect against mortality, while pulmonary conditions increase mortality risk, especially in older patients. These insights suggest that individualized risk assessments incorporating age and comorbidities are essential for managing COVID-19 outcomes.

Background and Objectives: The COVID-19 pandemic has had a significant impact on global health, with serious outcomes, such as lung damage, being major determinants of patient morbidity and mortality. Immunization has been essential in attenuating these outcomes. This study aimed to evaluate the impact of COVID-19 vaccination on disease severity, particularly focusing on pulmonary involvement, among hospitalized patients. Materials and Methods: A retrospective cohort study was conducted at Victor Babes Hospital, Timisoara, involving 3005 patients diagnosed with COVID-19 between December 2020 and March 2022. Patients were stratified into vaccinated and unvaccinated groups. Results: The study found that vaccinated patients had significantly lower rates of severe pulmonary involvement compared to unvaccinated patients. Specifically, only 24.24% of vaccinated patients experienced severe lung involvement, compared to 35.64% in the unvaccinated group (p < 0.001). Vaccinated individuals had shorter hospital stays (8.96 ± 6.40 days vs. 10.70 ± 6.29 days), but this difference was not statistically significant (p = 0.219). Additionally, chronic pulmonary diseases and stroke were less prevalent among vaccinated patients, highlighting the protective effect of vaccination. Conclusions: COVID-19 vaccination significantly reduces the severity of disease, particularly in preventing severe pulmonary involvement, which is a major determinant of patient outcomes. These findings underscore the importance of ongoing vaccination efforts and the need for booster doses to maintain immunity, especially as new variants emerge. The study supports the continued prioritization of vaccination in public health strategies to mitigate the long-term impact of COVID-19.

This study represents our second investigation into NIPT, involving a more extensive patient cohort with a specific emphasis on the high-risk group. The high-risk group was subsequently divided into two further groups to compare confirmed cases versus unconfirmed via direct methods. The methodology encompassed the analysis of 1400 consecutive cases from a single genetic center in western Romania, where NIPT was used to assess the risk of specific fetal chromosomal abnormalities. All high-risk cases underwent validation through direct analysis of fetal cells obtained via invasive methods, including chorionic villus sampling and amniocentesis. The confirmation process utilized QF-PCR, karyotyping, and SNP-Array methods customized to each case. Results: A high risk of aneuploidy at NIPT was identified in 36 out of 1400 (2.57%) cases and confirmed in 28 cases. The study also detected an increased risk for copy number variations (CNVs) in 1% of cases, confirmed in two instances involving one large microdeletion and one large microduplication. Trisomy 21 was the exclusive anomaly where NIPT confirmed all cases with identified risk. High-risk NIPT results which were not validated by invasive methods, were classified as false positives; parents in these cases determined to continue the pregnancy. In conclusion, NIPT can serve as a screening method for all pregnancies; however, in high-risk cases, an invasive confirmation test is strongly recommended.

Background and Objectives: Non-invasive prenatal testing (NIPT) has been confirmed as the most accurate screening test for trisomies 21, 18, 13, sex chromosomes aneuploidies and several microdeletions. This study aimed to assess the accuracy of cell free DNA testing based on low-level whole-genome sequencing to screen for these chromosomal abnormalities and to evaluate the clinical performance of NIPT. Materials and Methods: 380 consecutive cases from a single genetic center, from Western Romania were included in this retrospective study. Cell-free nucleic acid extraction from maternal blood, DNA sequencing and analysis of sequenced regions were performed by BGI Hong Kong and Invitae USA to determine the risk of specific fetal chromosomal abnormalities. In high-risk cases the results were checked by direct analysis of fetal cells obtained by invasive methods: 6 chorionic villus sampling and 10 amniocenteses followed by combinations of QF-PCR, karyotyping and aCGH. Results: NIPT results indicated low risk in 95.76% of cases and high risk in 4.23%. Seven aneuploidies and one microdeletion were confirmed, the other results were found to be a false-positive. A gestational age of up to 22 weeks had no influence on fetal fraction. There were no significant differences in fetal fraction across the high and low risk groups. Conclusions: This is the first study in Romania to report the NIPT results. The confirmation rate was higher for autosomal aneuploidies compared to sex chromosome aneuploidies and microdeletions. All cases at risk for trisomy 21 were confirmed. Only one large fetal microdeletion detected by NIPT has been confirmed. False positive NIPT results, not confirmed by invasive methods, led to the decision to continue the pregnancy. The main limitation of the study is the small number of patients included. NIPT can be used as a screening method for all pregnancies, but in high-risk cases, an invasive confirmation test was performed.