Search

ABSTRACT Silver‐Russell syndrome (SRS) is an imprinting disorder mainly characterized by pre‐ and postnatal growth restriction. Most SRS cases are due to 11p15.5 loss of methylation (11p15.5 LOM) or maternal uniparental disomy of chromosome 7 [UPD(7)mat], but several patients remain molecularly undiagnosed. This study describes the molecular investigation of children with a clinical diagnosis or suspicion of SRS at a tertiary center specialized in growth disorders. Thirty‐nine patients were evaluated with multiplex ligation‐dependent probe amplification, chromosomal microarray and/or massively parallel sequencing. The most common result was 11p15.5 LOM (n = 17; 43.5%), followed by UPD(7)mat (n = 2; 5.1%). Additionally, we found maternal duplications of the imprinting centers in 11p15.5 (n = 2; 5.1%), and genetic defects in SRS‐causing genes (IGF2 and HMGA2) (n = 3; 7.7%; two mutations and one deletion). Alternative molecular diagnoses included UPD(14)mat (n = 1; 2,6%), UPD(20)mat (n = 1;2,6%), copy number variants (n = 2; 5.1%), and mutations in genes associated with other growth disorders (n = 4; 10.3%), leading to diagnoses of Temple syndrome, Mulchandani‐Bhoj‐Conlin syndrome, IGF‐1 resistance (IGF1R), Bloom syndrome (BLM), Gabriele‐De Vries syndrome (YY1), Intellectual developmental disorder autosomal dominant 50 with behavioral abnormalities (NAA15), and Intellectual developmental disorder 64 (ZNF292). These findings underscore the importance of establishing the molecular diagnosis of SRS and its differential diagnoses to guide appropriate management and genetic counseling. [ABSTRACT FROM AUTHOR]

Objectives Patients and Methods Results Conclusion To investigate the impact of the revised papillary renal cell carcinoma (PRCC) classification and evaluate its validity with regard to oncological outcome stratification.Identifying 527 patients with PRCC who underwent surgical resection from 1995 to 2022, a tissue microarray was constructed for immunohistochemical and molecular characterisation. Re‐classification according to the World Health Organization (WHO) 2022 criteria and nuclear grading according to the WHO/International Society of Urological Pathologists criteria were done. In addition to the revised subtype, alleged clinicopathological prognosticators were analysed with respect to progression‐free (PFS) and cancer‐specific survival (CSS).Initially, 247 (46.9%) cases were Type 1, 234 (44.4%) were Type 2, and 46 (8.7%) were papillary not‐otherwise‐specified. According to the revised criteria, 29.9% of Type 1 and 57.7% of Type 2 PRCC cases were re‐classified. Re‐classified from Type 1 included more indolent tumours while from Type 2 PRCC many molecularly defined tumours were newly identified. After re‐classification, still 373 tumours remained with distinct histomorphological features of Type 1 (254 [70%]) and Type 2 (119 [42.2%]) PRCC. Furthermore, significant differences in survival outcomes were obtained when the revised criteria was used particularly for tumours of ≤4 cm. For a median (interquartile range) follow‐up of 79 (38.2–132.8) months, the 5‐year PFS was 97% for Type 1, 80% for Type 2, 75% for transcription factor for immunoglobulin heavy‐chain enhancer 3 (TFE3)‐rearranged, and 43.5% for fumarate hydratase‐deficient RCC. No disease progression was observed in patients with papillary renal neoplasm with reverse polarity.The revised WHO 2022 classification enhanced prognostic accuracy for PRCC particularly for small tumours. Retaining previous subtypes may confer further clinical as well as prognostic value. [ABSTRACT FROM AUTHOR]

Most follicular cell-derived differentiated thyroid carcinomas are regarded as low-risk neoplasms prompting conservative therapeutic management. Here, we provide consensus recommendations reached by a multidisciplinary group of endocrinologists, medical oncologists, pathologists, radiation oncology specialists, a surgeon and a medication reimbursement specialist, addressing more challenging forms of this malignancy, focused on radioactive iodine (RAI)-resistant or -refractory differentiated thyroid carcinoma (RAIRTC). In this document we highlight clinical, radiographic, and molecular features providing the basis for these management plans. We distinguish differentiated thyroid cancers associated with more aggressive behavior from thyroid cancers manifesting as poorly differentiated and/or anaplastic carcinomas. Treatment algorithms based on risk-benefit assessments of different multimodal therapy approaches are also discussed. Given the scarcity of data supporting management of this rare yet aggressive disease entity, these consensus recommendations provide much needed guidance for multidisciplinary teams to optimally manage RAIRTC. [ABSTRACT FROM AUTHOR]

Summary Vacuoles, E1 enzyme, X‐linked, autoinflammatory, somatic (VEXAS) is a haemato‐inflammatory syndrome genetically defined by somatic mutations in the X‐linked UBA1 gene, typically Val/Thr/Leu substitutions at the Met41 hotspot. Clinical manifestations are heterogeneous and refractory to most haemato‐rheumatological treatments. To date, no guidelines exist for the management of VEXAS, and scarce is the evidence on methodology and clinical significance of longitudinal UBA1 clonal burden evaluation upon therapy. Here, we validated a method to quantify UBA1 clonal burden and explored its applicability in patients with VEXAS. Given the different treatment interactions, droplet digital polymerase chain reaction (ddPCR) may allow for informed therapeutic decisions and implementation of personalized strategies. [ABSTRACT FROM AUTHOR]

Next-generation sequencing technologies have not only defined a breakthrough in medical genetics, but also been able to enter routine clinical practice to determine individual genetic susceptibilities. Modern technological developments are routinely introduced to genetic analysis overtaking the established approaches, potentially raising a number of challenges. To what extent is the advantage of new methodologies in synthetic metrics, such as precision and recall, more important than stability and reproducibility? Could differences in the technical protocol for calling variants be crucial to the diagnosis and, by extension, the patient's treatment strategy? A regulatory review process may delay the incorporation of potentially beneficial technologies, resulting in missed opportunities to make the right medical decisions. On the other hand, a blind adoption of new technologies based solely on synthetic metrics of precision and recall can lead to incorrect conclusions and adverse outcomes for the specific patient. Here, we use the example of a patient with a WHO-diagnosed desmoplastic/nodular SHH-medulloblastoma to explore how the choice of DNA variant search protocol affects the genetic diagnostics outcome. [ABSTRACT FROM AUTHOR]

Cornelia de Lange syndrome (CdLS, OMIM #122470, #300590, #300882, #610759, and #614701) is a rare congenital disorder that affects the development of multiple organs and is characterized by physical abnormalities and cognitive and behavioral disabilities. Its molecular basis is mainly based on alterations in genes encoding structural and regulatory proteins related to the cohesin complex. Moreover, other transcriptional regulatory factors have been linked to this syndrome. However, additional causative genes are still unknown, since many patients still lack a molecular diagnosis. Herein, we describe a case with multiple affected family members presenting with an intragenic duplication in the AFF2 gene. The direct tandem intragenic duplication of exons 10, 11 and 12 was detected through high-resolution array Comparative Genomic Hybridization and next-generation sequencing technologies. Confirming the X-linked inheritance pattern, the duplication was found in the patient, his mother and his maternal aunt affected (dizygotic twins). Targeted sequencing with Oxford Nanopore Technologies revealed an aberrant transcript which is predominantly expressed in the patient and his aunt. Along with these results, a significant reduction in AFF2 gene expression levels was detected in these two individuals. Clinically both subjects exhibit a classic CdLS phenotype, whereas the mother is mostly unaffected. Consistent with the phenotypical differences observed between the mother and the aunt, there is a marked difference in X-inactivation patterns skewing. Given the crucial role of AFF2 in transcriptional regulation, it is not surprising that AFF2 variants can give rise to CdLS phenotypes. Therefore, the AFF2 gene should be considered for the molecular diagnosis of this syndrome. [ABSTRACT FROM AUTHOR]

Pancreatic cancer (PC) is one of the deadliest malignant tumors worldwide, known as the 'king of cancers due to its insidious onset, high malignancy, and high mortality rate. PC is highly malignant and progresses rapidly, but its onset is insidious with atypical early symptoms, making it difficult to detect early lesions through conventional imaging studies. It is usually only discovered when symptoms like jaundice, abdominal and back pain occur. Surgical resection is currently the only curative option for PC. However, due to the difficulty in early diagnosis, the majority of patients are already in the middle to late stages at the time of diagnosis, missing the opportunity for surgery. Studies have confirmed that the progression of pancreatic cancer is relatively slow, with the initial tumor cells requiring at least 15 years to gain metastatic ability. Therefore, timely detection of pancreatic cancer through tumor markers could significantly improve the survival rate of patients. The most widely used and diagnostically valuable tumor marker in clinical practice is Carbohydrate antigen 199 (CA199). However, due to about 3% ~ 7% of pancreatic cancer patients being Lewis antigen-negative blood types and not expressing CA19-9, its sensitivity is only 79% ~ 81%, which does not provide good efficacy for the diagnosis of pancreatic cancer. Pancreatic juice, as a fluid near the tumor, has attracted much attention as a good source of tumor-related biomarkers. Many studies have confirmed the accuracy of using proteins, DNA, and exosomes in pancreatic juice for the diagnosis of pancreatic cancer, showing great prospects for pancreatic juice as a source of tumor markers for the diagnosis of pancreatic cancer. Therefore, this thesis reviews the efficacy of pancreatic juice as a specimen for the diagnosis of pancreatic cancer. [ABSTRACT FROM AUTHOR]

Background: We have recently constructed a DNA methylation classifier that can discriminate between pancreatic ductal adenocarcinoma (PAAD) liver metastasis and intrahepatic cholangiocarcinoma (iCCA) with high accuracy (PAAD-iCCA-Classifier). PAAD is one of the leading causes of cancer of unknown primary and diagnosis is based on exclusion of other malignancies. Therefore, our focus was to investigate whether the PAAD-iCCA-Classifier can be used to diagnose PAAD metastases from other sites. Methods: For this scope, the anomaly detection filter of the initial classifier was expanded by 8 additional mimicker carcinomas, amounting to a total of 10 carcinomas in the negative class. We validated the updated version of the classifier on a validation set, which consisted of a biological cohort (n = 3579) and a technical one (n = 15). We then assessed the performance of the classifier on a test set, which included a positive control cohort of 16 PAAD metastases from various sites and a cohort of 124 negative control samples consisting of 96 breast cancer metastases from 18 anatomical sites and 28 carcinoma metastases to the brain. Results: The updated PAAD-iCCA-Classifier achieved 98.21% accuracy on the biological validation samples, and on the technical validation ones it reached 100%. The classifier also correctly identified 15/16 (93.75%) metastases of the positive control as PAAD, and on the negative control, it correctly classified 122/124 samples (98.39%) for a 97.85% overall accuracy on the test set. We used this DNA methylation dataset to explore the organotropism of PAAD metastases and observed that PAAD liver metastases are distinct from PAAD peritoneal carcinomatosis and primary PAAD, and are characterized by specific copy number alterations and hypomethylation of enhancers involved in epithelial-mesenchymal-transition. Conclusions: The updated PAAD-iCCA-Classifier (available at https://classifier.tgc-research.de/) can accurately classify PAAD samples from various metastatic sites and it can serve as a diagnostic aid. [ABSTRACT FROM AUTHOR]

CRISPR-Cas12a system, characterized by its precise sequence recognition and cleavage activity, has emerged as a powerful and programmable tool for molecular diagnostics. However, current CRISPR-Cas12a-based nucleic acid detection methods, particularly microRNA (miRNA) detection, necessitate additional bio-engineering strategies to exert control over Cas12a activity. Herein, we propose an engineered target-responsive hairpin DNA activator (TRHDA) to mediate forward tearing protospacer activated CRISPR-Cas12a system, which enables direct miRNA detection with high specificity and sensitivity. Target miRNA specifically binding to hairpin DNA can drive forward tearing protospacer in the stem sequence of hairpin structure, facilitating the complementarity between crRNA spacer and protospacer to activate Cas12a. Upon the hairpin DNA as input-responsive activator of Cas12a, a universal biosensing method enables the multiple miRNAs (miR-21, let-7a, miR-30a) detection and also has exceptional capability in identifying single-base mismatches and distinguishing homologous let-7/miR-30 family members. Besides, TRHDA-mediated Cas12a-powered biosensing has realized the evaluation of miR-21 expression levels in diverse cellular contexts by intracellular imaging. Considering the easy programmability of hairpin DNA in responsive region, this strategy could expand for the other target molecules detection (e.g., proteins, micromolecules, peptides, exosomes), which offers significant implications for biomarkers diagnostics utilizing the CRISPR-Cas12a system toolbox. [ABSTRACT FROM AUTHOR]

The accurate prenatal diagnosis of skeletal anomaly (SKA) using prenatal imaging alone remains challenging. We aimed to investigate the efficacy of whole-exome sequencing (WES) in the prenatal molecular genetic diagnosis of skeletal system abnormalities, with or without additional ultrasound anomalies. All fetuses with SKA were subjected to sequential genetic tests, and after excluding fetal chromosomal abnormalities and clinically significant copy number variations (CNVs) consistent with the observed phenotype, the affected fetuses were further subjected to WES. The clinical features of fetal SKA were collected, and the results of molecular genetic testing and perinatal outcomes were analyzed. Following negative routine genetic test results of the 78 fetuses, trio-WES was conducted for 73 fetuses, and fetus-only WES (single WES) was performed for five fetuses due to parental refusal. Fetal skeletal system abnormalities in our cohort were subdivided into seven groups: 39 (50%) had short long bones, 14 (17.9%) had abnormal limb morphology, 4 (5.1%) had polydactyly, 4 (5.1%) had the absence of the radius tibia or tibiofibula, 5 (6.4%) had spine anomalies, 6 (7.7%) had strephenopodia, and 6 (7.7%) had multiple deformities. In total, we identified the molecular diagnoses for 32/78 fetuses with SKAs, and confirmed 41 pathogenic/likely pathogenic variants in 28 genes, including nine novel variants in our cohort. The overall diagnostic rate was 41% (32/78). Our findings demonstrate that WES can greatly improve the genetic diagnostic rate of fetal SKAs following routine genetic testing, which can comprehensively guide perinatal management and help assess the risk of recurrence in future pregnancies. Our data also provide a basis for the association between the SKA phenotype and related genotypes and expand the spectrum of fetal SKA phenotypes and related genes. [ABSTRACT FROM AUTHOR]

Monkeypox virus clade I is endemic in several central African countries and characterised by an increase in disease severity and mortality. Since October, 2023, a large-scale mpox outbreak has emerged in DR Congo, and in March, 2024, the first individuals with mpox were reported outside the endemic areas in Republic of the Congo. We aimed to provide insight into the epidemic by sequencing samples obtained from individuals with mpox in Republic of the Congo. In this cross-sectional, descriptive study, samples were collected from individuals with suspected mpox between Jan 15 and April 8, 2024, in Brazzaville, Pointe-Noire, Likouala, Cuvette-Centrale, and Plateaux (Republic of the Congo). Blood samples, skin or oropharyngeal swabs, or skin crusts were obtained for molecular diagnosis via real-time PCR. Monkeypox virus sequences were obtained and analysed using newly established nanopore sequencing methodology and bioinformatic pipeline. The sequences obtained were aligned and used to construct a maximum likelihood phylogenetic tree using IG-TREE. 61 samples were collected from individuals with suspected mpox, 31 of which were positive for monkeypox virus and were included in our analysis (four positive samples were excluded due to unavailability of epidemiological data or insufficient biological material). Individuals who tested positive for monkeypox virus were from Cuvette-Centrale (19 [61%] of 31), Likouala (eight [26%]), and Pointe-Noire (four [13%]). 20 (65%) were male and 11 (35%) were female. Phylogenetic analysis of sequences showed two major clusters within clade Ia. One cluster was made up of four sequences from this study clustering with two monkeypox virus sequences from the current DR Congo outbreak, three older sequences from Central African Republic sequenced between 2017 and 2018, and seven sequences from DR Congo sequenced in 2006–07 and 2022. The second cluster was made up of 16 sequences from this study clustering with sequences from the current DR Congo outbreak. In addition, sequences from Republic of the Congo show multiple phylogenetic positioning suggesting the occurrence of multiple co-circulating strains in the human population. Our findings suggest that multiple monkeypox virus strains are co-circulating in the human population, highlighting the need for implementation of expanded mpox surveillance, especially in countries bordering DR Congo and Republic of the Congo, in combination with control measures focused on containing the current outbreaks in these countries to prevent escalation into a larger-scale epidemic. Intramural Research Program of the National Institute of Allergy and Infectious Diseases at the National Institutes of Health. [ABSTRACT FROM AUTHOR]

ABSTRACT Background Methods Results Conclusions There is growing evidence that the Spitz group of melanocytic neoplasms should be restricted to those harboring kinase receptor fusions and HRAS mutations/11p15 amplification. The presence of genomic alterations characteristic of conventional melanomas (BRAF and NRAS mutations) precludes a diagnosis of a Spitz neoplasm. It is often challenging to distinguish Spitz neoplasms from conventional melanomas with spitzoid morphology on histopathological grounds alone.We report a series of 70 consecutive melanocytic tumors in which targeted sequencing was indicated to distinguish Spitz from spitzoid neoplasms and to classify Spitz neoplasms along the biological spectrum.Final diagnoses incorporating molecular results included 12 conventional melanomas (nine of which with NRAS mutations), five Spitz melanomas, 35 atypical Spitz tumors, eight Spitz nevi, three melanocytic tumors with a MAP2K1 mutation, and seven desmoplastic Spitz nevi/tumors. There were significant discrepancies between initial diagnoses and final diagnoses after incorporating molecular results in 24 (34%) cases, including nine conventional melanomas favored to be Spitz neoplasms and nine Spitz neoplasms favored to be conventional melanomas.It is often not possible to reliably distinguish Spitz neoplasms from spitzoid melanocytic tumors without identifying their driver genomic alterations. Applying next‐generation sequencing in diagnostically problematic tumors improves diagnostic accuracy. [ABSTRACT FROM AUTHOR]

As more patients receive genome-wide sequencing, the number of individuals diagnosed with multiple monogenic conditions is increasing. We sought to investigate the relative phenotypic contribution of dual diagnoses using both manual curation and computational approaches. First, we computed 1,003,236 semantic similarity scores for all possible pairs of 1,417 genes in the Developmental Disorder Gene2Phenotype (DDG2P) database using Human Phenotype Ontology terms. Next, for 62 probands with two molecular diagnoses in the Deciphering Developmental Disorders study, we computed semantic similarity scores between the probands' phenotypes and DDG2P phenotypes associated with the two disorders and compared the results with manual attribution of proband phenotypes to none, one, or both of the genes. We found a spectrum of phenotypic similarity for dual diagnoses, both across all DDG2P genes and within dual diagnosed probands, from phenotypically distinct through blended to indistinguishable conditions. Pairwise semantic similarity scores between two DDG2P genes were a good predictor of the extent of phenotypic blending observed in probands. Dual diagnoses involving genes linked with synergistic phenotypes can result in more extreme presentations while those involving antagonistic phenotypes have spuriously high pairwise semantic similarity scores despite a potentially milder atypical presentation. We suggest that the phenotypic contribution of two molecular diagnoses may contain discrete, synergistic, or antagonistic elements. Conceptual recognition of this phenotypic spectrum is important for making a final clinico-molecular diagnosis and providing accurate genetic counseling. Computational and manual assessment of dual diagnoses in developmental disorders reveals a spectrum of phenotypic similarity from distinct through blended to indistinguishable in which synergistic or antagonistic elements may result in absent or extreme phenotypes. [ABSTRACT FROM AUTHOR]

Digital nucleic acid quantification has emerged as an exceptionally valuable technique in molecular diagnosis, capturing significant attention in the biomedical field. This study introduces a novel multi‐volume digital loop‐mediated isothermal amplification (dLAMP) chip that offers an extensive detection range and user‐friendly operation. The chip consists of 1024 large chambers with the volume of 7.2 nL and 6400 small chambers with the volume of 0.064 nL. The ingenious combination of chambers with volume difference beyond a 100‐fold on the single chip enables a broad dynamic range exceeding 105 for nucleic acid quantification. Furthermore, this chip incorporates an automated strategy, enabling the rapid loading and partitioning of the reaction mixture into subvolumes within 4 min. In order to evaluate the quantitative capability of the multi‐volume dLAMP chip, a 10‐fold serial dilution of the β‐actin DNA template is measured. Additionally, the chip is tested for β‐actin DNA concentration in plasma samples from colorectal cancer patients and healthy individuals. By providing enhanced automation, wide dynamic range, and improved accuracy, this chip paves the way for broader applications and the advancement of biomedical research fields. [ABSTRACT FROM AUTHOR]

Introduction: Hearing loss is a frequent sensory impairment type in humans, with about 50% of prelingual cases being attributed to genetic factors. Autosomal recessive hearing loss (ARHL) exhibits great locus heterogeneity and is responsible for 70%-80% of hereditary nonsyndromic cases. Methods: A total of 90 unrelated Brazilian individuals were selected for having hearing loss of presumably autosomal recessive inheritance, either born from consanguineous marriages or belonging to families with two or more affected individuals in the sibship and most cases were of normal hearing parents. In all cases, common pathogenic variants in GJB2 (c.35delG), GJB6 [del(GJB6-D13S1830) and del(GJB6-D13S1854)] and MT-RNR1 (m.1555A>G) were discarded and most were previously assessed by complete Sanger sequencing of GJB2. Their genetic material was analyzed through next-generation sequencing, targeting 99 hearing loss-related genes and/or whole exome sequencing. Results: In 32 of the 90 probands (36,7%) causative variants were identified, with autosomal recessive inheritance confirmed in all, except for two cases due to dominant variants (SIX1 and P2RX2). Thirty-nine different causative variants were found in 24 different known hearing loss-associated genes, among which 10 variants are novel, indicating wide genetic heterogeneity in the sample, after exclusion of common pathogenic variants. Despite the genetic heterogeneity, some genes showed greater contribution: GJB2, CDH23, MYO15A, OTOF, and USH2A. Conclusion: The present results confirmed that next-generation sequencing is an effective tool for identifying causative variants in autosomal recessive hearing loss. To our knowledge, this is the first report of next-generation sequencing being applied to a large cohort of pedigrees with presumable autosomal recessive hearing loss in Brazil and South America. [ABSTRACT FROM AUTHOR]

Introduction: Hearing loss is a frequent sensory impairment type in humans, with about 50% of prelingual cases being attributed to genetic factors. Autosomal recessive hearing loss (ARHL) exhibits great locus heterogeneity and is responsible for 70%–80% of hereditary nonsyndromic cases. Methods: A total of 90 unrelated Brazilian individuals were selected for having hearing loss of presumably autosomal recessive inheritance, either born from consanguineous marriages or belonging to families with two or more affected individuals in the sibship and most cases were of normal hearing parents. In all cases, common pathogenic variants in GJB2 (c.35delG), GJB6 [del(GJB6-D13S1830) and del(GJB6-D13S1854)] and MT-RNR1 (m.1555A>G) were discarded and most were previously assessed by complete Sanger sequencing of GJB2. Their genetic material was analyzed through next-generation sequencing, targeting 99 hearing loss-related genes and/or whole exome sequencing. Results: In 32 of the 90 probands (36,7%) causative variants were identified, with autosomal recessive inheritance confirmed in all, except for two cases due to dominant variants (SIX1 and P2RX2). Thirty-nine different causative variants were found in 24 different known hearing loss-associated genes, among which 10 variants are novel, indicating wide genetic heterogeneity in the sample, after exclusion of common pathogenic variants. Despite the genetic heterogeneity, some genes showed greater contribution: GJB2 , CDH23 , MYO15A , OTOF , and USH2A. Conclusion: The present results confirmed that next-generation sequencing is an effective tool for identifying causative variants in autosomal recessive hearing loss. To our knowledge, this is the first report of next-generation sequencing being applied to a large cohort of pedigrees with presumable autosomal recessive hearing loss in Brazil and South America. [ABSTRACT FROM AUTHOR]

Introduction: Magnetic particle imaging (MPI), a radiation-free, dynamic, and targeted imaging technique, has gained significant traction in both research and clinical settings worldwide. Signal-to-noise ratio (SNR) is a crucial factor influencing MPI image quality and detection sensitivity, and it is affected by ambient noise, system thermal noise, and the magnetization response of superparamagnetic nanoparticles. Therefore to address the high amplitude system and inherent thermal noise present in conventional MPI systems is essential to improve detection sensitivity and imaging resolution. Method: This study introduces a novel open-loop, narrow-band MPI signal acquisition system based on mixed-frequency harmonic magnetization response. Allowing superparamagnetic nanoparticles to be excited by low frequency, high amplitude magnetic fields and high frequency, low amplitude magnetic fields, the excitation coil generates a mixed excitation magnetic field at a mixed frequency of 8.664 kHz (f H + 2 f L ), and the tracer of superparamagnetic nanoparticles can generate a locatable superparamagnetic magnetization signal with rich harmonic components in the mixed excitation magnetic field and positioning magnetic field. The third harmonic signal is detected by a Gradiometer coil with high signal-to-noise ratio, and the voltage cloud image is formed. Result: The experimental results show that the external noise caused by the excitation coil can be effectively reduced from 12 to about 1.5 μV in the imaging area of 30 mm × 30 mm, which improves the stability of the detection signal of the Gradiometer coil, realizes the detection of high SNR, and makes the detection sensitivity reach 10 μg Fe. By mixing excitation, the total intensity of the excitation field is reduced, resulting in a slight improvement of the resolution under the same gradient field, and the spatial resolution of the image reconstruction is increased from 2 mm under the single frequency excitation (20.7 kHz) in the previous experiment to 1.5 mm under the mixed excitation (8.664 kHz). Conclusions: These experimental results highlight the effectiveness of the proposed open-loop narrowband MPI technique in improving signal detection sensitivity, achieving high signal-to-noise ratio detection and improving the quality of reconstructed images by changing the excitation magnetic field frequency of the excitation coil, providing novel design ideas and technical pathways for future MPI systems. [ABSTRACT FROM AUTHOR]

Babesia spp. and Theileria spp. are tick-borne apicomplexan protozoa that can cause disease in animals and humans. Deer are considered reservoirs for a wide variety of Piroplasmida species, including some potentially zoonotic. This study aimed to investigate the occurrence and genetic diversity of piroplasmids in wild deer sampled in four Brazilian states (São Paulo, Mato Grosso do Sul, Paraná and Goiás). For this purpose, extracted DNA samples from 181 deer buffy coat samples (138 Blastocerus dichotomus, 26 Subulo gouazoubira, 4 Mazama jucunda, 3 Mazama rufa and 10 Ozotocerus bezoarticus) were subjected to a nested PCR (nPCR) assay based on the 18S rRNA gene in order to perform a screening for piroplasmids and characterized based on the near-complete 18S rRNA, hsp70 and cox-3 genes. As a result, 75.14% (136/181) samples were positive for piroplasmids. Of these, 108 (79.41%), 101 (74.26%) and 67 (49.26%) were positive to near complete 18S rRNA, hsp70 and cox-3 genes, respectively. Phylogenetic analyses based on three molecular markers showed similar topology to each other. All sequences obtained in the present study were positioned into the Theileria sensu stricto clade, forming a distinct clade, albeit close to T. cervi. Most sequences grouped together into a large clade divided into subclades, which were often related to deer genus/species, showing that Theileria lineages seemed to show specificity according to deer genus/species. Two 18S rRNA sequences (one obtained from S. gouazoubira and another from M. jucunda) were positioned into a different clade, apart from other sequences detected in this study, indicating that different species of Theileria occur in deer from Brazil. Two subclusters were observed in the phylogenetic analysis based on the hsp70 gene: the first containing only sequences detected in marsh deer and the second grouping sequences detected in brocket deer (Mazama spp. and S. gouazoubira). The latter was also divided into smaller clades that grouped Theileria genotypes according to deer species (M. jucunda, M. rufa and S. gouazoubira). This study provides the first molecular evidence of Theileria infection in M. jucunda, as well as co-infection by distinct Theileria (sub)species/genotypes in the same deer was evidenced. Finally, this study expanded the knowledge on the diversity of Theileria spp. infecting deer from South America. [ABSTRACT FROM AUTHOR]

A novel approach is introduced using nanoplasmonic microarray–based solid-phase recombinase polymerase amplification (RPA) that offers high sensitivity and multiplexing capabilities for gene detection. Nanoplasmonic microarrays were developed through one-step immobilization of streptavidin/biotin primers and fine-tuning the amplicon size to achieve high plasmon-enhanced fluorescence (PEF) on the nanoplasmonic substrate, thereby improving sensitivity. The specificity and sensitivity of solid-phase RPA on nanoplasmonic microarrays was evaluated in detecting E, N, and RdRP genes of SARS-CoV-2. High specificity was achieved by minimizing primer-dimer formation and employing a stringent washing process and high sensitivity obtained with a limit of detection of four copies per reaction within 30 min. In clinical testing with nasopharyngeal swab samples (n = 30), the nanoplasmonic microarrays demonstrated a 100% consistency with the PCR results for detecting SARS-CoV-2, including differentiation of Omicron mutations BA.1 and BA.2. This approach overcomes the sensitivity issue of solid-phase amplification, as well as offers rapidity, high multiplexing capabilities, and simplified equipment by using isothermal reaction, making it a valuable tool for on-site molecular diagnostics. [ABSTRACT FROM AUTHOR]

Background: Pancreatic cancer (PC) is one of the most aggressive types of cancer. Despite advances in molecular diagnostics, PC diagnosis relies on imaging technologies and morphological assessment of fine needle aspirates (FNAs). MicroRNA (miRNA) involvement in PC pathogenesis and potential diagnostics application have been suggested, albeit current supporting evidence is lacking. Here, we investigated the association of selected miRNAs with PC incidence and clinical characteristics. Methods: Fold expression of miR‐216a‐3p, ‐217‐5p, ‐221‐3p, ‐222‐3p, and miR‐196a‐5p was assessed in 73 PC FNA cell‐block sections and 6 healthy pancreas tissues using Taqman advanced miRNA assays. Potential miRNA targets were ascertained using immunocytochemistry. Results: miR‐196a‐5p was upregulated in PC compared to healthy pancreatic tissue (β = −0.05, 95% CI: −0.065 – (−0.035); p < 0.001). miR‐221‐3p and miR‐222‐3p fold expression were strongly correlated (r = 0.897, p < 0.001), whereas miR‐196a‐5p fold expression correlated with that of miR‐221‐3p (r = 0.688, p < 0.001) and miR‐222‐3p (r = 0.489, p < 0.001). Moreover, miR‐196a‐5p fold expression positively correlated with tumor stage (r = 0.32, p = 0.034). miR‐217‐5p fold expression inversely correlated with KRAS expression (r = ‐0.69, p = 0.0027). Conclusion: Our study shows miR‐196a‐5p has reasonable specificity to PC and thus may have diagnostic and prognostic potential in PC as proposed in the literature. Moreover, KRAS and NFKBIA may be potential targets for miR‐217‐5p and miR‐196a‐5p, respectively. Thus, these miRNAs may be involved in tumor progression and may have valuable applications in novel therapeutics or treatment monitoring. [ABSTRACT FROM AUTHOR]

On stage, and in real life, timing is critical for success. Theranostic cancer care epitomizes the central role of timing in the evolution of efficacious molecular targeted radioligand therapy and its incorporation into routine clinical practice of oncology. Nuclear medicine has returned to its therapeutic roots, having been founded as a medical specialty, over three-quarters of a century ago, with radioiodine therapy of thyroid cancer. The very recent oncologist acceptance of 68Ga/177Lu/225Ac-PSMA effectiveness in treating prostate cancer has re-established the role of the physician in nuclear medicine. This article addresses various important issues in respect of timing related to this resurgence. Training of the required new workforce in technical -omics expertise and physicianly virtues is an urgent priority. Precision in radioligand therapy requires definition of individual radiation absorbed dose (Gy) to tumor and to critical normal organs, preferably prospectively. It is time to abandon one-size-fits-all administration of fixed activities (GBq) in arbitrary cycle intervals and duration. The time has also come to design combination sequenced theranostic-immuno-chemotherapeutic approaches to metastatic cancer to address unmet needs, particularly in pancreatic carcinoma; exploiting the potential of new fibroblast activation protein inhibitor radioligands targeting the tumor microenvironment. Public perception of all things "nuclear," including nuclear medicine, has recently recovered from the general opprobrium and radiophobia of the last half-century. Nuclear is the new green. At last, there have arisen propitious circumstances for the future development of theranostics: The timing is right, now. [ABSTRACT FROM AUTHOR]

(1) Background: Group A Streptococcal (GAS) pharyngitis is common, resulting in numerous ambulatory visits. Accurate diagnosis is challenging. This study evaluated the clinical utility, cost, and performance of a nucleic acid amplification test (NAAT) for GAS detection, comparing it to a rapid antigen detection test (RADT) and throat culture. Additionally, we assessed the diagnostic stewardship related to these testing methods to ensure appropriate antibiotic use in clinical practice. Methods: Between November 2022 and February 2023, 82 throat swabs were analyzed, with McIsaac clinical scores calculated for each. The Abbott ID NOW STREP A 2 NAAT and Sekisui Diagnostics' OSOM® STREP A RADT were performed, followed by bacterial culture. Diagnostic performance was compared using culture as the gold standard. Results: Of the 82 samples, 28 (34.14%) tested positive for pathogenic germs, primarily Streptococcus pyogenes (92.85%). RADTs showed a sensitivity of 80.76% and a specificity of 100%, while NAATs demonstrated a sensitivity of 100% and specificity of 96.42%. Cost analysis indicated the need for reimbursement adjustments to optimize NAAT's economic benefits. Clinical data indicated that symptoms alone were insufficient for reliable diagnosis. Conclusions: This study confirmed the superior sensitivity of Abbott's Strep A2 NAAT over RADT. Given the Belgian guidelines against routine antibiotic treatment for pharyngitis and considering local treatment recommendations and cost, implementing NAAT for GAS detection in Belgian laboratories is less beneficial. However, the role of NAAT in supporting antimicrobial stewardship by ensuring appropriate antibiotic use remains significant. [ABSTRACT FROM AUTHOR]

Background: Smooth Muscle Tumor of Uncertain Malignant Potential (STUMP) is a poorly studied neoplasm that does not fulfill the definition of either leiomyoma or leiomyosarcoma. STUMP symptoms are indistinguishable from those of benign lesions; it has no specific biochemical markers or ultrasound presentations. The management of this type of tumor is particularly challenging due to significant heterogeneity in its behavior and the lack of clear guidelines; moreover, the lesion may recur after excision. Case Report: We report on a case of a 42-year-old patient diagnosed with a STUMP. The preliminary diagnosis was a submucous leiomyoma, which was removed hysteroscopically due to menorrhagia resulting in anemia. The histopathological examination of the resected myoma pointed to the diagnosis of STUMP. The hysterectomy was performed as the patient had completed her reproductive plans. There were no complications. The patient is currently recurrence-free after a 9-month follow-up. Discussion and Conclusions: The care of a patient diagnosed with STUMP requires a personalized approach and the cooperation of various medical disciplines, including molecular diagnostics, imaging techniques, and minimally invasive surgery. Management of STUMP must consider the patient's plans for childbearing. All cases of tumors with "uncertain malignant potential" are a challenge in the context of patient-physician communication. [ABSTRACT FROM AUTHOR]

Biochemical phenotyping has been the milestone for diagnosing and managing patients affected by inborn errors of intermediary metabolism (IEiM); however, identifying the genotype responsible for these monogenic disorders greatly contributes to achieving these goals. Herein, whole-exome sequencing (WES) was used to determine the genotypes of 95 unrelated Mexican pediatric patients suspected of having IEiM. They were classified into those bearing specific biochemical abnormalities (Group 1), and those presenting unspecific biochemical profiles (Group 2). The overall concordance between the initial biochemical diagnosis and final genotypic diagnoses was 72.6% (N = 69/95 patients), with the highest concordance achieved in Group 1 (91.3%, N = 63/69), whereas the concordance was limited in Group 2 (23.07%). This finding suggests that previous biochemical phenotyping correlated with the high WES diagnostic success. Concordance was high for urea cycle disorders (94.1%) and organic acid disorders (77.4%). The identified mutational spectrum comprised 83 IEiM-relevant variants (pathogenic, likely pathogenic, and variants of uncertain significance or VUS), including three novel ones, distributed among 29 different genes responsible for amino acid, organic acid, urea cycle, carbohydrate, and lipid disorders. Inconclusive WES results (7.3%, N = 7/95) relied on monoallelic pathogenic genotypes or those involving two VUS for autosomal-recessive IEiMs. A second monogenic disease was observed in 10.5% (N = 10/95) of the patients. According to the WES results, modifications in treatment had to be made in 33.6% (N = 32/95) of patients, mainly attributed to the presence of a second monogenic disease, or to an actionable trait. This study includes the largest cohort of Mexican patients to date with biochemically suspected IEiM who were genetically diagnosed through WES, underscoring its importance in medical management. [ABSTRACT FROM AUTHOR]

Background/Objectives: Rapid and accurate pathogen identification is essential for the proper management of patients with infectious gastroenteritis, as well as for a better control of disease outbreaks. This observational, non-interventional, single-site study evaluated the diagnostic accuracy of LiquidArray® Gastrointestinal VER 1.0, a multiplex PCR syndromic panel capable of detecting up to 26 clinically relevant enteropathogens. Methods: Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and likelihood ratio (LR) were evaluated using stool samples from 1512 patients with suspected gastroenteritis and were compared to seven competitor assays. Results: LiquidArray® Gastrointestinal VER 1.0 showed a very low invalid rate (0.5% at initial testing, down to 0% after repeat) and high sensitivity (>90% for most detected targets) and specificity (>99% for all detected targets). Accordingly, the PPV and NPV were high (>90% for most targets and >99% for all targets, respectively). The analytical performance of LiquidArray® Gastrointestinal VER 1.0 was also excellent as to co-amplification capability, cross-reactivity and assay precision. Conclusions: This study demonstrates the excellent clinical performance of LiquidArray® Gastrointestinal VER 1.0 and its suitability for implementation in clinical routine for the rapid and accurate diagnosis of infectious gastroenteritis. [ABSTRACT FROM AUTHOR]

Simple Summary: Molecular testing is becoming the standard of care across multiple tumor types in oncology. In non-small cell lung cancer (NSCLC), the uptake of molecular testing with next-generation sequencing (NGS) has been earlier than in other fields due to the clinical relevance of the molecular results. There are multiple approaches to NGS, ranging from sending testing internally, often reflexively, to referring externally to limited or comprehensive panels. This study aims to identify the differences in time to results, time to treatment, molecular alterations, and clinical outcomes for different molecular workflows in NSCLC. The results may impact the workflows for testing in a variety of cancers. Purpose: Broad-based molecular testing with next-generation sequencing (NGS) is now the standard of care in advanced non-small cell lung cancer (NSCLC). Two approaches to molecular testing are (1) reflexive testing at pathologic NSCLC confirmation, often using an in-house molecular panel, and (2) send-out testing to private vendors, ordered by a clinician. This study explored the outcomes with reflex versus send-out testing. Methods: A retrospective chart review was conducted of patients diagnosed with de novo stage IV NSCLC in 2019 and 2020 at three hospitals in the same system, one academic hospital (Northwestern Memorial Hospital, or NMH) utilizing reflex, in-house NGS, and two community-based hospitals (Central DuPage Hospital, or CDH, and Delnor, or D) sending out tissue samples for testing. The outcomes assessed were the time from biopsy to results, biopsy to treatment, the incidence of first-line targetable mutations and the use of first-line targeted therapies, and overall survival. Results: In total, 191 patients met the inclusion criteria, 85 at NMH, 106 at CDH + D, and in total, 131 in 2019 and 60 in 2020. The time to results was significantly shorter with reflexive NGS when compared with send-out testing; the time to treatment was also shorter but not statistically significant. At CDH + D, the time to results was significantly shorter with a limited panel than with comprehensive testing, but the time to treatment was similar. NGS testing rates were 95% at NMH and 84.5% at CDH + D (p = 0.009), with 31.0% at NMH receiving 1L targeted therapies versus 20.8% at CDH + D (p = 0.08). In 2019, the median time from biopsy to treatment was 35 days at NMH and 38 days at CDH and Delnor; in 2020, time to treatment was 26 days and 37 days, respectively. Overall survival trended longer in 2020 relative to 2019 independent of site. Conclusion: Reflexive NGS testing is associated with a shorter time to actionable results and higher rates of first-line targetable mutations than send-out testing. In practices with send-out testing, limited panels had slightly faster turnaround times but no difference in time to treatment. If resources allow, reflexive NGS should be considered in healthcare systems for patients with NSCLC. [ABSTRACT FROM AUTHOR]

Simple Summary: STING agonists face challenges in breast cancer treatment due to their lower accumulation in tumors and rapid clearance from the body, resulting in a short duration of therapeutic effect. Novel phase-transitional liposomes loaded with STING agonists have been developed to overcome these limitations and are specifically designed for low-intensity focused ultrasound (LIFU)-assisted molecular imaging and precise treatment of breast cancer. With the assistance of LIFU, iRGD-modified liposomes (a targeting peptide) undergo a phase transition into microbubbles, leading to enhanced ultrasound molecular imaging of tumors and facilitating breast cancer immunotherapy by releasing STING agonists from the ultrasound-targeted liposomes. Background: Pharmacologically targeting the STING pathway offers a novel approach to cancer immunotherapy. However, small-molecule STING agonists face challenges such as poor tumor accumulation, rapid clearance, and short-lived effects within the tumor microenvironment, thus limiting their therapeutic potential. To address the challenges of poor specificity and inadequate targeting of STING in breast cancer treatment, herein, we report the design and development of a targeted liposomal delivery system modified with the tumor-targeting peptide iRGD (iRGD-STING-PFP@liposomes). With LIFU irradiation, the liposomal system exploits acoustic cavitation, where gas nuclei form and collapse within the hydrophobic region of the liposome lipid bilayer (transient pore formation), which leads to significantly enhanced drug release. Methods: Transmission electron microscopy (TEM) was used to investigate the physicochemical properties of the targeted liposomes. Encapsulation efficiency and in vitro release were assessed using the dialysis bag method, while the effects of iRGD on liposome targeting were evaluated through laser confocal microscopy. The CCK-8 assay was used to investigate the toxicity and cell growth effects of this system on 4T1 breast cancer cells and HUVEC vascular endothelial cells. A subcutaneous breast cancer tumor model was established to evaluate the tumor-killing effects and therapeutic mechanism of the newly developed liposomes. Results: The liposome carrier exhibited a regular morphology, with a particle size of 232.16 ± 19.82 nm, as indicated by dynamic light scattering (DLS), and demonstrated low toxicity to both HUVEC and 4T1 cells. With an encapsulation efficiency of 41.82 ± 5.67%, the carrier exhibited a slow release pattern in vitro after STING loading. Targeting results indicated that iRGD modification enhanced the system's ability to target 4T1 cells. The iRGD-STING-PFP@liposomes group demonstrated significant tumor growth inhibition in the subcutaneous breast cancer mouse model with effective activation of the immune system, resulting in the highest populations of matured dendritic cells (71.2 ± 5.4%), increased presentation of tumor-related antigens, promoted CD8+ T cell infiltration at the tumor site, and enhanced NK cell activity. Conclusions: The iRGD-STING-PFP@liposomes targeted drug delivery system effectively targets breast cancer cells, providing a new strategy for breast cancer immunotherapy. These findings indicate that iRGD-STING-PFP@liposomes could successfully deliver STING agonists to tumor tissue, trigger the innate immune response, and may serve as a potential platform for targeted immunotherapy. [ABSTRACT FROM AUTHOR]

Simple Summary: Serous ovarian carcinoma (SOC) is the most common type of ovarian cancer, but predicting its behaviour and patient outcomes has been challenging. This research is the first to comprehensively assess histopathological features such as lymphovascular space invasion, tumour budding, the tumour–stroma ratio, the stromal type, microvessel density, tumour-infiltrating lymphocytes, and tertiary lymphoid structures in the same cohort of both primary and metastatic SOC cases. By simultaneously evaluating these features, our study provides new insights into the tumour microenvironment and its role in disease progression. This comprehensive approach highlights the prognostic value of these features and offers a straightforward method for assessing tumour aggressiveness in routine clinical practice. These findings could lead to better risk stratification and personalised treatment strategies for SOC patients, particularly in settings where access to advanced molecular testing is limited. Background/Objectives: Serous ovarian carcinoma (SOC) is the most common subtype of epithelial ovarian cancer, with high-grade (HGSOC) and low-grade (LGSOC) subtypes presenting distinct clinical behaviours. This study aimed to evaluate histopathologic features in SOC, correlating these with prognostic outcomes, and explore the potential clinical implications. Methods: We analysed 51 SOC cases for lymphovascular space invasion (LVSI), tumour border configuration (TBC), microvessel density (MVD), tumour budding (TB), the tumour–stroma ratio (TSR), the stromal type, tumour-infiltrating lymphocytes (TILs), and tertiary lymphoid structures (TLSs). A validation cohort of 54 SOC cases from The Cancer Genome Atlas (TCGA) was used for comparison. Results: In the discovery set, significant predictors of aggressive behaviour included LVSI, high MVD, high TB, and low TILs. These findings were validated in the validation set where the absence of TLSs, lower peritumoural TILs, immature stromal type, and low TSR were associated with worse survival outcomes. The stromal type was identified as an independent prognostic predictor in SOC across both datasets. Inter-observer variability analysis demonstrated substantial to almost perfect agreement for these features, ensuring the reproducibility of the findings. Conclusions: The histopathological evaluation of immune and stromal features, such as TILs, TLSs, TB, TSR, and stromal type, provides critical prognostic information for SOC. Incorporating these markers into routine pathological assessments could enhance risk stratification and guide treatment, offering practical utility, particularly in low-resource settings when molecular testing is not feasible. [ABSTRACT FROM AUTHOR]

Objective: This study aims to evaluate the clinical management and effectiveness of metagenomic next-generation sequencing (mNGS) in patients with pyogenic spinal infections. Methods: We conducted a retrospective review of 17 patients diagnosed with pyogenic spinal infections and treated at our institution between October 2022 and February 2024. The cohort included 8 males and 9 females, with a mean age of 63.59 ± 10.18 years (range: 41–71 years). The infections comprised 9 epidural abscesses, 6 intervertebral space infections, and 2 deep abscesses. All patients underwent open surgical procedures and mNGS-based bacterial identification using intraoperative pus or tissue specimens, in addition to conventional blood bacterial cultures. Clinical outcomes were assessed using CRP, PCT, WBC inflammatory markers, and VAS scores postoperatively. Results: All 17 patients with pyogenic spinal infections underwent open surgery and mNGS bacterial detection at our institution. Among the 17 patients, mNGS yielded positive results in 14 cases (82.4%), significantly higher than the 5.9% positivity rate of conventional bacterial cultures (p < 0.001). The mNGS test time was notably shorter than conventional cultures (1.0 vs. 5.88 days, p < 0.001). Postoperative antibiotic therapy was adjusted based on mNGS findings. There were significant reductions in postoperative VAS, WBC, PCT, and CRP values compared to preoperative levels (p < 0.01). Conclusion: Metagenomic next-generation sequencing is effective in managing pyogenic spinal infections by facilitating rapid and sensitive detection of pathogens. This technique improves the timeliness and accuracy of diagnosis, highlighting its potential for broader clinical use. [ABSTRACT FROM AUTHOR]

Context.--: Mesothelioma is an uncommon tumor that can be difficult to diagnose. Objective.--: To provide updated, practical guidelines for the pathologic diagnosis of mesothelioma. Data sources.--: Pathologists involved in the International Mesothelioma Interest Group and others with expertise in mesothelioma contributed to this update. Reference material includes peer-reviewed publications and textbooks. Conclusions.--: There was consensus opinion regarding guidelines for (1) histomorphologic diagnosis of mesothelial tumors, including distinction of epithelioid, biphasic, and sarcomatoid mesothelioma; recognition of morphologic variants and patterns; and recognition of common morphologic pitfalls; (2) molecular pathogenesis of mesothelioma; (3) application of immunohistochemical markers to establish mesothelial lineage and distinguish mesothelioma from common morphologic differentials; (4) application of ancillary studies to distinguish benign from malignant mesothelial proliferations, including BAP1 and MTAP immunostains; novel immunomarkers such as Merlin and p53; fluorescence in situ hybridization (FISH) for homozygous deletion of CDKN2A; and novel molecular assays; (5) practical recommendations for routine reporting of mesothelioma, including grading epithelioid mesothelioma and other prognostic parameters; (6) diagnosis of mesothelioma in situ; (7) cytologic diagnosis of mesothelioma, including use of immunostains and molecular assays; and (8) features of nonmalignant peritoneal mesothelial lesions. [ABSTRACT FROM AUTHOR]

Context.--Salivary gland tumors are rare tumor types for which the molecular understanding has resulted in a rapid expansion and shuffling of entities. These changes are reflected in the 5th edition World Health Organization Classification of Head and Neck Tumours (WHO 5th edition), although many nuances still remain. Objective.--To review how molecular alterations have helped recategorize, justify, and reinstate entities into our lexicon as well as defining interrelationships between categories, new entities, and subtypes. Furthermore, newer theranostic applications to molecular phenotype will be summarized. Data Sources.--World Health Organization Classification of Head and Neck Tumours (WHO 3rd through 5th editions), literature review, and personal and institutional experience. Conclusions.--Molecular alterations have helped reclassify, retain, and create new categories by augmenting rather than replacing standard criteria. Key entities that have emerged include sclerosing polycystic adenoma, microsecretory adenocarcinoma, and mucinous adenocarcinoma. Molecular phenotypes solidify the range of morphology in established entities such as mucoepidermoid carcinoma and facilitate connectivity between entities. Molecular characteristics now allow for targeted therapeutic approaches for secretory carcinoma and adenoid cystic carcinoma. [ABSTRACT FROM AUTHOR]

HERC5, a vital protein in the HERC family, plays crucial roles in immune response, cancer progression, and antiviral defense. This bioinformatic study comprehensively assessed HERC5's significance across various malignancies by analyzing its gene expression, immune and molecular subtype expressions, target proteins, biological functions, and prognostic and diagnostic values in pan‐cancer. We further examined its correlation with clinical features, co‐expressed and differentially expressed genes, and prognosis in clinical subgroups, focusing on endometrial cancer (UCEC). Our findings showed that HERC5 RNA is expressed at low levels in most cancers and significantly differs across immune and molecular subtypes. HERC5 accurately predicts cancer and correlates with most cancer prognoses. In UCEC, HERC5 was significantly associated with age, hormonal status, clinical stage, treatment status, and metastasis. Elevated HERC5 expression was linked to worse progression‐free interval, disease‐specific survival, and overall survival in UCEC, particularly in diverse clinical subgroups. Significant differences in HERC5 expression were also observed in various human cancer cell line validations. In summary, HERC5 may be a critical biomarker for pan‐cancer prognosis, progression, and diagnosis, as well as a promising new target for cancer therapy. [ABSTRACT FROM AUTHOR]

Identification of the molecular culprits of allergic reactions leveraged molecular allergology applications in clinical laboratory medicine. Molecular allergology shifted the focus from complex, heterogeneous allergenic extracts, e.g. pollen, food, or insect venom, towards genetically and immunologically defined proteins available for in vitro diagnosis. Molecular allergology is a precision medicine approach for the diagnosis, stratification, therapeutic management, follow-up and prognostic evaluation of patients within a large range of allergic diseases. Exclusively available for in vitro diagnosis, molecular allergology is nonredundant with any of the current clinical tools for allergy investigation. As an example of a major application, discrimination of genuine sensitization from allergen cross-reactivity at the molecular level allows the proper targeting of the culprit allergen and thus dramatically improves patient management. This review aims at introducing clinical laboratory specialists to molecular allergology, from the biochemical and genetic bases, through immunological concepts, to daily use in the diagnosis and management of allergic diseases. [ABSTRACT FROM AUTHOR]

Due to the major improvements in the hardware and image reconstruction algorithms, positron emission tomography/magnetic resonance imaging (PET/MR) is now a reliable state-of-the-art hybrid modality in medical practice. Currently, it can provide a broad range of advantages in preclinical and clinical imaging compared to single-modality imaging. In the second part of this review, we discussed the further clinical applications of PET/MR. In the chest, PET/MR has particular potential in the oncology setting, especially when utilizing ultrashort/zero echo time MR sequences. Furthermore, cardiac PET/MR can provide reliable information in evaluating myocardial inflammation, cardiac amyloidosis, myocardial perfusion, myocardial viability, atherosclerotic plaque, and cardiac masses. In gastrointestinal and hepato-pancreato-biliary malignancies, PET/MR is able to precisely detect metastases to the liver, being superior over the other imaging modalities. In genitourinary and gynaecology applications, PET/MR is a comprehensive diagnostic method, especially in prostate, endometrial, and cervical cancers. Its simultaneous acquisition has been shown to outperform other imaging techniques for the detection of pelvic nodal metastases and is also a reliable modality in radiation planning. Lastly, in haematologic malignancies, PET/MR can significantly enhance lymphoma diagnosis, particularly in detecting extra-nodal involvement. It can also comprehensively assess treatment-induced changes. Furthermore, PET/MR may soon become a routine in multiple myeloma management, being a one-stop shop for evaluating bone, bone marrow, and soft tissues. [ABSTRACT FROM AUTHOR]

Copyright of Die Urologie is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The article focuses on the challenges faced by oncologists in keeping up with the rapid advancements in molecular testing and its integration into clinical practice. Topics include the role of molecular diagnostics in cancer treatment, the importance of circulating tumor DNA (ctDNA) in early detection and therapy selection, and the practical challenges related to the adoption of molecular testing, such as educational gaps and reimbursement issues.

The article focuses on the ethical, clinical, and regulatory challenges of using molecular diagnostic tests, such as circulating tumor DNA (ctDNA), in cancer care. Topics include the evolving role of ctDNA testing in detecting minimal residual disease, the ethical considerations surrounding early adoption of tests with uncertain clinical benefits, and the responsibility of oncologists in guiding patients through emerging technologies.

Until the early 2000s, advanced or metastatic non–small cell lung cancer (NSCLC) was treated as a single disease with all histologic subtypes treated alike with standard chemotherapy agents. Over the past two decades, the treatment paradigms for advanced NSCLC have changed dramatically with the discovery of multiple targeted therapies that are now approved for the treatment of NSCLC tumors with specific oncogene drivers or molecular alterations. Molecular testing has become integrated and critical for the clinical management of advanced NSCLC. The discovery and success of these targeted therapies have reshaped the classification of NSCLC on the basis of molecular classification and enabled a personalized approach in thoracic oncology. In this review, we discuss recent developments in the molecular profiling of NSCLC, and approved and emerging targeted therapies for the treatment of NSCLC. In this review, we discuss recent developments in the molecular profiling of non–small cell lung cancer (NSCLC), and approved and emerging targeted therapies for the treatment of NSCLC. [ABSTRACT FROM AUTHOR]

Over the next few years, the analysis of circulating tumor DNA (ctDNA) through liquid biopsy is expected to enter clinical practice and revolutionize the approach to biomarker testing and treatment selection in GI cancers. In fact, growing evidence support the use of ctDNA testing as a noninvasive, effective, and highly specific tool for molecular profiling in GI cancers. Analysis of blood ctDNA has been investigated in multiple settings including early tumor detection, minimal residual disease evaluation, tumor diagnosis and evaluation of prognostic/predictive biomarkers for targeted treatment selection, longitudinal monitoring of treatment response, and identification of resistance mechanisms. Here, we review the clinical applications, advantages, and limitations of ctDNA profiling for precision oncology in GI cancers. [ABSTRACT FROM AUTHOR]

The article focuses on the challenges faced by oncologists in interpreting complex molecular data for precision oncology and providing optimal treatment. Topics include the rapid advancement of molecular testing, the effectiveness of molecular tumor boards in clinical decision-making, and efforts to expand community access to such resources.

As molecular imaging use expands for patients with breast cancer, it is important for breast radiologists to have a basic understanding of molecular imaging, including PET. Although breast radiologists may not directly interpret such studies, basic knowledge of molecular imaging will enable the radiologist to better direct diagnostic workup of patients as well as discuss diagnostic imaging with the patient and other treating physicians. Several new tracers are now available to complement imaging glucose metabolism with FDG. Because it provides a noninvasive assessment of disease status across the whole body, PET offers specific advantages over tissue-based assays. Paired with targeted therapy, molecular imaging has the potential to guide personalized treatment of breast cancer, including guiding dosing during drug trials as well as predicting and assessing clinical response. This review discusses the current established applications of FDG, which remains the most widely used PET radiotracer for malignancy, including breast cancer, and highlights potential areas for expanded use based on recent research. It also summarizes research to date on the U.S. Food and Drug Administration (FDA)–approved PET tracer 16α-18F-fluoro-17β-estradiol (FES), which targets ER, including the current guidelines from the Society of Nuclear Medicine and Molecular Imaging on the appropriate use of FES-PET/CT for breast cancer as well as areas of active investigation for other potential applications. Finally, the review highlights several of the most promising novel PET tracers that are poised for clinical translation in the near future. [ABSTRACT FROM AUTHOR]

Multiple endocrine neoplasia (MEN) is a group of rare genetic diseases characterized by the occurrence of multiple tumors of the endocrine system in the same patient. The first MEN described was MEN1, followed by MEN2A, and MEN2B. The identification of the genes responsible for these syndromes led to the introduction of family genetic screening programs. More than twenty years later, not all cases of MENs have been resolved from a genetic point of view, and new clinicogenetic entities have been described. In this review, we will discuss the strategies and difficulties of genetic screening for classic and newly described MENs in a clinical setting, from limitations in sequencing, to problems in classifying variants, to the identification of new candidate genes. In the era of genomic medicine, characterization of new candidate genes and their specific tumor risk is essential for inclusion of patients in personalized medicine programs as well as to permit accurate genetic counseling to be proposed for families. [ABSTRACT FROM AUTHOR]

Background: Short stature is a complex disorder with phenotypic and genetic heterogeneity. This study aimed to investigate clinical phenotypes and molecular basis of a cohort of patients with short stature. Methods: Trio whole-exome sequencing (Trio-WES) was performed to explore the genetic aetiology and obtain a molecular diagnosis in twenty Chinese probands with syndromic and isolated short stature. Results: Of the twenty probands, six (6/20, 30%) patients with syndromic short stature obtained a molecular diagnosis. One novel COMP pathogenic variant c.1359delC, p.N453fs*62 and one LZTR1 likely pathogenic variant c.509G>A, p.R170Q were identified in a patient with short stature and skeletal dysplasia. One novel de novo NAA15 pathogenic variant c.63T>G, p.Y21X and one novel de novo KMT2A pathogenic variant c.3516T>A, p.N1172K was identified in two probands with short stature, intellectual disability and abnormal behaviours, respectively. One patient with short stature, cataract, and muscle weakness had a de novo POLG pathogenic variant c.2863 T>C, p.Y955H. One PHEX pathogenic variant c.1104G>A, p.W368X was identified in a patient with short stature and rickets. Maternal uniparental disomy 7 (mUPD7) was pathogenic in a patient with pre and postnatal growth retardation, wide forehead, triangular face, micrognathia and clinodactyly. Thirteen patients with isolated short stature had negative results. Conclusion: Trio-WES is an important strategy for identifying genetic variants and UPD in patients with syndromic short stature, in which dual genetic variants are existent in some individuals. It is important to differentiate between syndromic and isolated short stature. Genetic testing has a high yield for syndromic patients but low for isolated patients. [ABSTRACT FROM AUTHOR]

The burden of colorectal cancer (CRC) is high in the Asia‐Pacific region, and several countries in this region have among the highest and/or fastest growing rates of CRC in the world. A significant proportion of patients will present with or develop metastatic CRC (mCRC), and BRAFV600E‐mutant mCRC represents a particularly aggressive phenotype that is less responsive to standard chemotherapies. In light of recent therapeutic advances, an Asia‐Pacific expert consensus panel was convened to develop evidence‐based recommendations for the diagnosis, treatment, and management of patients with BRAFV600E‐mutant mCRC. The expert panel comprised nine medical oncologists from Australia, Hong Kong, Singapore, and Taiwan (the authors), who met to review current literature and develop eight consensus statements that describe the optimal management of BRAFV600E‐mutant mCRC in the Asia‐Pacific region. As agreed by the expert panel, the consensus statements recommend molecular testing at diagnosis to guide individualized treatment decisions, propose optimal treatment pathways according to microsatellite stability status, advocate for more frequent monitoring of BRAFV600E‐mutant mCRC, and discuss local treatment strategies for oligometastatic disease. Together, these expert consensus statements are intended to optimize treatment and improve outcomes for patients with BRAFV600E‐mutant mCRC in the Asia‐Pacific region. [ABSTRACT FROM AUTHOR]

Accurate detection of Helicobacter pylori and its antimicrobial resistance is essential for eradication of the infections. The aim of this study was to compare five different CE‐IVD marked assays in detection of H. pylori from 268 clinical stool samples. Samples were considered positive for H. pylori when at least three of the five tests were positive. Amplified IDEIA Hp StAR (Oxoid) and Premier Platinum HpSA PLUS (Meridian Bioscience Inc.) assays showed sensitivity of 100% [95% CI (confidence interval): 87–100] and LIAISON® Meridian H. pylori SA (DiaSorin) of 83.3% (95% CI: 66–93). Specificities of the assays were 94.5% (95% CI: 91–97), 95.4%; (95% CI: 92–97), and 97.1% (95% CI: 94–99) respectively. Amplidiag® H. pylori + ClariR (Mobidiag) assay showed 93.3% (95% CI: 78–99) and Allplex™ H. pylori & ClariR Assay (Seegene Inc.) 36.7% (95% CI: 22–55) sensitivity, while specificity of both was 97.9% (95% CI: 95–99). The Amplidiag® and Allplex™ assays concordantly detected clarithromycin resistance in positive for H. pylori samples. The Amplidiag® assay showed the highest accuracy, namely 97.4% (95% CI: 95–99). These data provide helpful information for planning laboratory diagnostics of H. pylori and detection of clarithromycin resistance from stool samples. [ABSTRACT FROM AUTHOR]