Search Results (86)

Background/Objectives: Gasdermin D (GSDMD) is a critical mediator of pyroptosis—an inflammatory form of programmed cell death increasingly implicated in tumor biology. Our objective was to evaluate the utility of GSDMD as a diagnostic and prognostic biomarker and to investigate its association with tumor burden and hematological parameters. Methods: We analyzed GSDMD expression levels in patients with adult-type diffuse gliomas compared to healthy controls and assessed correlations with tumor size, histological grade, hematological markers, and survival outcomes. Data was complemented by transcriptomic analysis from The Cancer Genome Atlas (TCGA). Diagnostic performance was assessed using ROC curve analysis. Results: GSDMD expression was significantly elevated in adult-type diffuse glioma patients and increased with tumor grade, suggesting an association with disease severity. A positive correlation was observed between GSDMD level and tumor size (R = 0.332; p = 0.01). ROC analysis showed moderate classification ability (AUC = 0.657) with high specificity (96%), supporting its diagnostic potential. Survival analysis showed that higher GSDMD expression was associated with reduced disease-specific survival. GSDMD also correlated positively with the erythrocyte parameter mean corpuscular hemoglobin (MCH, R = 0.34, p = 0.016) and negatively with the systemic inflammatory marker C-reactive protein (CRP, R = −0.32; p = 0.042). TCGA data showed no significant sex-related differences in GSDMD expression. Baseline characteristics such as age, BMI, and coagulation parameters were matched between patients and controls. Conclusions: GSDMD is significantly associated with astrocytoma severity, tumor size, and inflammatory status, with elevated expression indicating a worse prognosis. Its correlation with tumor grade, survival and high specificity in distinguishing patients from healthy individuals, underlines its promise as a clinically relevant, non-sex-specific biomarker for diagnosis and monitoring. Full article

Backgroud/Objectives: Pediatric central nervous system (CNS) tumors represent the second most common oncological disease in children. The purpose of this research was to analyze the course of surgical treatment for these tumors at the University Medical Centre Ljubljana between October 2018 and December 2025. Methods: A retrospective analysis of 110 patients was conducted, focusing on diagnostic accuracy and its correlation with surgical and neurological outcomes. Results: Over a seven-year period, 110 children were surgically treated, undergoing a total of 130 operative procedures. A calculated annual incidence of 3.8 cases per 100,000 was identified. The most common initial symptoms were headache (36.3%), vomiting (20%), and ataxia (17.2%). Tumors were localized supratentorially in 52.7% of cases, infratentorially in 30%, and along the spinal canal in 13.6%. The most frequent histopathological types were pilocytic astrocytomas/paediatric-type diffuse low-grade gliomas (18.1%), medulloblastomas (14.5%), and craniopharyngiomas (7.2%). Gross total resection was achieved in 60% of all procedures. Surgical complications occurred in 10.6% of cases, with a surgical mortality rate of 0.9%. Neurological deterioration occurred in 27.2% of cases, most commonly in the form of cerebellar mutism (8.1%). Diagnostic histology of medulloblastoma and infratentorial anatomical location were confirmed as critical prognostic markers for cerebellar mutism (p < 0.011) and shunt dependency (p = 0.0121). Conlcusions: This study confirms that treatment outcomes in the Slovenian tertiary center are comparable to international standards. Integrative diagnostic strategies significantly refine surgical planning and prognostic assessment, optimizing long-term morbidity management. Full article

Glioblastoma, isocitrate dehydrogenase (IDH1/2) wild-type (IDH-wildtype), is one of the most aggressive and malignant tumors of the central nervous system, characterized by rapid growth, pronounced cellular heterogeneity, and an exceptionally poor prognosis. The median survival time for patients with glioblastoma, IDH-wildtype, is approximately 15 months after diagnosis, and current multimodal treatment strategies remain largely ineffective. This review focuses on contemporary pharmacotherapeutic approaches used in the management of glioblastoma, IDH-wildtype, including temozolomide-based chemotherapy, corticosteroids for edema control, and antiangiogenic therapy in recurrent disease, with particular emphasis on their clinical efficacy and limitations. In addition, the review discusses emerging targeted therapeutic strategies developed for IDH-mutant diffuse gliomas, which represent a biologically distinct disease entity. Particular attention is given to ivosidenib, a selective inhibitor of mutant IDH1, currently evaluated for the treatment of astrocytoma, IDH-mutant, grade 4. Its epigenetic mechanism of action, involving inhibition of the oncometabolite 2-hydroxyglutarate (2-HG), is outlined, along with preliminary clinical evidence suggesting potential to delay disease progression. Finally, innovative drug-delivery technologies designed to overcome the blood–brain barrier are briefly discussed as complementary strategies that may enhance the efficacy of both conventional and targeted therapies. Overall, future advances in the treatment of diffuse gliomas will likely depend on the integration of molecularly targeted agents, predictive biomarkers, and advanced delivery platforms aimed at improving patient survival and quality of life. Full article

Background/Objectives: The 2021 WHO Classification of Central Nervous System (CNS) tumors emphasizes the integration of molecular data with histopathological features. Lower-grade gliomas (LGGs) represent a heterogeneous group of neoplasms with variable clinical behavior. This study aimed to explore the molecular landscape of a single-institution series of LGGs using targeted next-generation sequencing (NGS). Methods: Eleven adult patients diagnosed with LGG between 2015 and 2024 at Cattinara University Hospital (Trieste, Italy) were retrospectively analyzed. DNA and RNA were extracted from formalin-fixed, paraffin-embedded (FFPE) tissue and analyzed using the TruSight Oncology 500 panel (Illumina). Mutational, amplification, and transcriptomic profiles were evaluated. Results: IDH1 mutations were the most frequent alteration (75%), commonly co-occurring with TP53 and ATRX mutations, consistent with the canonical IDH-mutant astrocytoma profile. CDK4 amplification was found in four cases, while MYCN amplification and MET amplification were each identified in isolated cases. Two diffuse IDH-wild-type gliomas displayed aggressive clinical courses and shorter survival, and one was reclassified as glioblastoma (grade 4) based on EGFR amplification. The transcriptome analysis revealed heterogeneous expression signatures and distinct clustering of IDH1/ATRX-mutant tumors. Conclusions: Targeted NGS confirmed the key molecular features of diffuse gliomas and enabled precise WHO 2021 classification even in archival FFPE samples. Despite the exploratory nature of the analysis on a small population, the study underscores the biological and transcriptional heterogeneity of LGGs and highlights the limitations of tumor-only sequencing approaches. Broader genomic profiling and matched normal controls are warranted to refine the interpretation of rare or non-canonical variants. Full article

Background: The androgen receptor (AR) is a ligand-dependent transcription factor of the nuclear steroid receptor superfamily, implicated in the pathogenesis of various solid tumors. The AR gene, located on chromosome Xq11–12, is accompanied by several X-linked genes that modulate AR expression and function, including FLNA, UXT, and members of the melanoma antigen gene (MAGE) family (MAGEA1, MAGEA11, MAGEC1, MAGEC2). While the AR has been investigated in multiple tumor types, its role in adult-type diffuse gliomas remains largely unexplored. Here, we characterized AR protein expression and the promoter methylation status of the AR and associated regulatory genes in adult-type diffuse gliomas. Methods: A retrospective analysis was conducted on 50 patients with adult-type diffuse gliomas, including IDH-mutant gliomas (grades 2–4) and IDH-wildtype glioblastomas (GBMs), classified according to the 2021 WHO criteria. AR nuclear expression was assessed by immunohistochemistry (IHC). Methylation-specific PCR and quantitative DNA methylation analyses were employed to evaluate promoter methylation of the AR and selected co-regulatory genes. Results: AR nuclear positivity correlated significantly with male sex (p = 0.04) and higher tumor grade, with the highest expression in IDH-wildtype GBMs (p = 0.04). In IDH-mutant gliomas, AR immunoreactivity was more prevalent in astrocytomas than in 1p/19q codeleted oligodendrogliomas (p = 0.02). AR expression was associated with unmethylated MGMT promoter status (p = 0.02). DNA methylation analysis revealed AR gene hypomethylation in tumors displaying nuclear AR positivity and in IDH-wildtype GBMs (Kruskal–Wallis p < 0.05). Additionally, methylation patterns of AR co-regulators located on the X chromosome suggest epigenetic regulation of AR signaling in gliomas. Conclusions: The findings reveal distinct AR pathway activation patterns in adult-type diffuse gliomas, particularly IDH-wildtype GBMs, suggesting that further exploration of antiandrogen therapies is warranted. Full article

Background/Objectives: Astrocytoma, IDH-mutant, CNS WHO grade 3, is a diffuse glioma with poor prognosis, molecularly defined by IDH mutations and frequently co-occurring TP53 and ATRX alterations. This study aimed to delineate the genomic landscape and identify clinically relevant molecular features of astrocytoma, IDH-mutant, CNS WHO grade 3 using this resource. Methods: Patients in the American Association for Cancer Research Project Genomics Evidence Neoplasia Information Exchange (AACR Project GENIE) database were selected based on histological diagnosis of “anaplastic astrocytoma”, confirmed IDH1/2 mutation, and exclusion of CDKN2A/B homozygous deletions. We analyzed frequencies of somatic mutations, copy number alterations (CNAs), structural variants (SVs), assessed co-occurrence/exclusivity patterns, and explored associations with available demographic and limited survival data. Results: The most common somatic mutations were in IDH1 (98.0%), TP53 (94.8%), and ATRX (55.2%). The observed ATRX mutation frequency was lower than some historical reports (e.g., ~86%). Other recurrent alterations included phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) (6.9%), Notch receptor 1 (NOTCH1) (6.9%), and platelet-derived growth factor receptor alpha (PDGFRA) (mutations 4.3%; CNAs also observed). Conclusions: This study provides a comprehensive genomic characterization of astrocytoma, IDH-mutant, CNS WHO grade 3 using the AACR GENIE database, confirming core mutational signatures while also highlighting potential variations in alteration frequencies, such as for ATRX. The findings establish a valuable real-world genomic benchmark for this tumor type, while promoting the need for continued data integration with robust clinical outcomes to identify actionable prognostic and therapeutic targets. Full article

Diffuse astrocytoma is a primary brain tumor known for its gradual and diffuse infiltration into the surrounding brain tissue. Given this characteristic, the investigation of the peritumoral region holds potential biological and clinical relevance. In this study, ion mobility spectrometry mass spectrometry (IMS MS) was optimized and applied for the first time for the analysis of gangliosides present in the peritumoral tissue of diffuse astrocytoma. Ganglioside profiling and structural characterization were conducted using high-resolution nanoelectrospray ionization (nanoESI) IMS MS, along with tandem mass spectrometry (MS/MS) via low-energy collision-induced dissociation (CID) in the negative ion mode. Using IMS MS-based separation and screening, we observed a greater diversity of ganglioside species in the peritumoral tissue than previously reported. Notably, an elevated expression was detected for several species, including GT1(d18:1/18:0), GT1(d18:1/20:0), GM2(d18:1/16:2), GD1(d18:1/16:0), GD2(d18:1/20:0), Fuc-GT3(d18:1/24:4), and Fuc-GD1(d18:1/18:2). Although preliminary, these observations prompt consideration of whether these species could be implicated in processes such as microenvironmental modulation, tumor cell infiltration and invasion, maintenance of cellular interactions, or regulation of immune responses. Additionally, their potential utility as biomarkers may merit further exploration. In the subsequent phase of the study, structural analysis using IMS MS, CID tandem MS, and fragmentation data supported the identification of GT1b(d18:1/20:0) isomer in the peritumoral tissue. However, given the exploratory nature of the study and reliance on limited sampling, further investigation across broader sample sets is necessary to extend these findings. Full article

Background/Objectives: Adult-type diffuse gliomas, including astrocytoma and glioblastoma multiforme (GBM), are brain tumors with a very poor prognosis. While current treatment options for glioma patients are not providing satisfactory outcomes, research indicates that natural compounds could serve as alternative treatments. However, their low bioavailability requires nanotechnology solutions, such as liposomes. Methods: In this study, we propose the co-encapsulation of acteoside (ACT) with other natural compounds, cannabidiol (CBD) or naringenin (NG), in a cationic liposomal nanoformulation consisting of DOTAP and POPC lipids, which were prepared using the dry lipid film method. The liposomes were characterized by their physicochemical properties, including particle size, zeta potential, and polydispersity index (PDI), with additional analyses performed using ¹H Nuclear Magnetic Resonance (NMR). Furthermore, biological experiments were performed with U-87 MG astrocytoma and U-138 MG GBM cell lines and non-cancerous MRC-5 lung fibroblasts using the MTT assay and evaluating the expression of Bax and Bcl-xL to evaluate their potential as anticancer agents. Conclusions: The IC₅₀ values for the nanoformulations in U-138 MG cells at 48 h were 6 µM for ACT + CBD and 5 µM for ACT + NG. ACT and CBD or NG demonstrated a potential synergistic effect against GBM in a liposomal formulation. Notably, treatment with ACT + CBD (5 µM) and ACT + NG (5 µM) liposomal formulations significantly upregulated Bax protein level in U-138 cells at both 24 and 48 h. In parallel, ACT + CBD (5 µM) also modulated Bcl-xL protein level in both U-138 MG and U-87 MG cell lines at the same time points. The obtained nanoformulations were homogeneous and stable for 21 days, evidenced by a narrow particle size distribution, a low polydispersity index (PDI) < 0.3, and a positive zeta potential. Full article

Background/Objectives: This study evaluates intratumoral susceptibility signals (ITSS) as imaging markers for glioma grade prediction and their association with molecular and histopathologic features, in the context of the fifth edition of the World Health Organization Classification of Tumors of the Central Nervous System (WHO CNS5). Methods: We retrospectively analyzed patients with adult diffuse gliomas who underwent pretreatment magnetic resonance imaging. ITSS were semiquantitatively graded by two radiologists: grade 0 (no signal), grade 1 (1–5), grade 2 (6–10), and grade 3 (≥11). Relative cerebral blood volume (rCBV) and tumor volume were also obtained. Histopathologic features included tumor grade, Ki-67, mitotic count, necrosis, microvascular proliferation, and molecular alterations (isocitrate dehydrogenase [IDH], 1p/19q, cyclin-dependent kinase inhibitors 2A and 2B [CDKN2A/B], and p53). Regression models predicted tumor grade (low: 1–2, high: 3–4) using ITSS, tumor volume, and rCBV. ROC curves and diagnostic performance metrics were analyzed. Results: 99 patients were included. ITSS grading correlated with rCBV, tumor volume, mitotic count, Ki-67, and tumor grade (p < 0.001). ITSS grades 0–1 were associated with oligodendrogliomas and astrocytomas (p < 0.001), IDH mutations (p < 0.001), and 1p/19q co-deletions (p = 0.01). ITSS grades 2–3 were linked to glioblastomas (p < 0.001), necrosis (p < 0.001), microvascular proliferation (p < 0.001), and CDKN2A/B homozygous deletions (p = 0.02). Models combining ITSS with rCBV and volume showed AUC of 0.94 and 0.96 (p < 0.001), outperforming univariate models. Conclusions: Semiquantitative ITSS grading correlates with key histopathologic and molecular glioma features. Combined with perfusion and volumetric parameters, ITSS enhance non-invasive glioma grading, in alignment with WHO CNS5. Full article

Background: Polycystin 1 (PC1) and polycystin 2 (PC2) proteins are members of the transient receptor potential (TRP) channels family and are encoded from PKD1 and PKD2 genes, respectively. Until recently, the role of PKD1 and PKD2 has been associated with the pathogenesis of the kidney since mutations in these genes cause autosomal dominant polycystic kidney disease (ADPKD). Recent data implicates polycystins in the pathogenesis of solid tumors. In this aspect, the expression of PKD1 and PKD2 in human astrocytomas is largely unknown. The aim of the present research study was to investigate the expression of PKD1 and PKD2 in astrocytic tumors and correlate it with clinicopathological characteristics such as the grade of malignancy, age, and gender of the patients. Methods: A total of 70 cases—corresponding to 8 grade II (diffuse fibrillary astrocytomas), 12 grade III (anaplastic astrocytomas), and 50 grade IV (glioblastomas multiforme)—were examined. The mRNA expression levels of PKD1 and PKD2 were determined through molecular qRT-PCR analysis using the relative quantification ΔΔCt method and the expression of PC1 and PC2 was detected through immunohistochemistry using the semi-quantitative H-score system. Results: Increased levels of PKD1 and PKD2 in astrocytomas were found compared with that of a normal brain (p < 0.05). Glioblastomas demonstrated the greatest increase in PKD1 and PKD2 expression compared to other grades of malignancy (p < 0.05). The same pattern of expression showed PC1 and PC2 proteins. A significant correlation between PKD1 and PKD2 as well as PC1 and PC2 expressions was found (p < 0.05). Although no association was detected between PC1 or PC2 and Ki67 expression (p > 0.05), a significant correlation between PC1 and p53 immunoexpressions, in grade III and between PC2 and p53 immunoexpressions, in grade II astrocytomas (p < 0.01) has emerged. PC1 expression was correlated with age of the patients (p < 0.05). PKD1 and PKD2 expression were negatively correlated with the prognosis of glioma patients. Conclusions: The results of this study indicate the potential involvement of polycystins in the pathogenesis of astrocytomas. However, further research is required to fully understand the mechanisms that these molecules are implicated. Full article

Background/Objectives: The purpose of this study was to evaluate the performance of diffusion kurtosis imaging (DKI), neurite orientation dispersion and density imaging (NODDI), and diffusion microstructure imaging (DMI) in differentiating molecular subtypes of adult-type gliomas. Methods: Standardized MRI was performed and evaluated in 59 patients with adult-type glioma. DKI, NODDI, and DMI parameter values were quantitatively evaluated in ROIs in contrast-enhancing/solid tumor tissue and five concentric shells with peritumoral tissue. DKI, NODDI, and DMI parameters of (i) glioblastomas, Isocitrate dehydrogenase (IDH) wildtype; (ii) astrocytomas, IDH mutant; and (iii) oligodendrogliomas, IDH mutant were compared with analysis of variance (ANOVA). Receiver operating characteristic curve (ROC) curve analysis was conducted to discriminate firstly between IDH mutant and IDH wildtype gliomas and then between IDH mutant astrocytomas and oligodendrogliomas. Results: Significant differences between the three aforementioned subtypes were found for the apparent diffusion coefficient (ADC) and mean kurtosis (MK) and again for the orientation dispersion index (ODI) and intra-axonal volume fraction (v-intra). The diagnostic accuracy depended on the distance to the contrast-enhancing/solid tumor tissue. Some NODDI and DMI parameters significantly predicted the IDH status and significantly discriminated between astrocytomas and oligodendrogliomas; however, ADC and MK showed the best prediction in both ROC analyses (maximum AUC 0.910 (CI 0.824–0.995)). Conclusions: The evaluation of peritumoral tissue can be a valuable procedure, while NODDI and DMI appear to be promising but are currently inferior to DKI in predicting glioma subtypes categorized according to the WHO 2021 classification. Full article

Background/Objectives: The non-invasive identification of glioma subtypes is useful for initial diagnosis, treatment planning, and follow-up. The aim of this study was to evaluate the performance of diffusion kurtosis imaging (DKI) and dynamic contrast-enhanced (DCE)-MRI in differentiating subtypes of adult-type diffuse gliomas. Methods: In a prospective multicenter study, standardized MRI was analyzed in 59 patients with adult-type diffuse glioma. DKI and DCE-MRI parameter values were quantitatively evaluated in ROIs of contrast-enhancing/solid tumor and four concentric shells of peritumoral tissue. The parameter means of glioblastomas, IDH wildtype; astrocytomas, IDH mutant; and oligodendrogliomas, IDH mutant were compared. Binary logistic regression analyses were performed to differentiate between IDH mutant and IDH wildtype gliomas and between IDH mutant astrocytomas and oligodendrogliomas. ROC curves were analyzed for each parameter and for combined regression. Results: Significant differences between the three aforementioned subtypes were found for the DKI and DCE-MRI parameters, depending on the distance to the tumor core. A combination of the parameters’ apparent diffusion coefficient (ADC) and fractional volume of extravascular extracellular space (ve) revealed the best prediction of IDH mutant vs. wildtype gliomas (AUC = 0.976 (0.943–1.000)) and astrocytomas vs. oligodendrogliomas (AUC = 0.840 (0.645–1.000)) with the lowest Akaike information criterion. Conclusions: The combined evaluation of DKI and DCE-MRI at different distances to the contrast-enhancing/solid tumor seems to be helpful in predicting glioma subtypes according to the WHO 2021 classification. Full article

Gliomas, ranging from low-grade pilocytic astrocytomas to highly malignant glioblastomas, are primary brain tumors that originate from neural or glial stem cells. Classified by the WHO into grades 1 to 4, these tumors exhibit varying prognoses, with oligodendrogliomas and astrocytomas having better and intermediate outcomes, respectively, while glioblastomas are associated with a poor prognosis. Despite advancements in molecular and genetic research that have improved diagnosis and the development of targeted therapies, treating high-grade gliomas remains a significant challenge due to their diffuse nature. In this context, lectins, carbohydrate-binding proteins, have shown promise as diagnostic and therapeutic agents for cancer, including gliomas. Plant lectins, particularly those from legumes, exhibit significant antiproliferative effects on glioma cells. These effects include decreased cell viability and migration, alongside the induction of autophagy and apoptosis, suggesting their potential as therapeutic agents. Although the mechanisms underlying these effects are not yet fully understood, molecular targets and pathways involved in the antiglioma activity of lectins have been identified. Key targets include matrix metalloproteinases (MMPs), epidermal growth factor receptor (EGFR), CD98 (xc^- system), AMPA receptor, and CD73. This review focuses on the antiglioma potential of legume lectins, their applications, and the main molecular targets based on their functions, structures, and associated molecular mechanisms. Full article

Adult-type diffuse gliomas are characterized by inevitable recurrence and very poor prognosis. Novel treatment options, including multimodal drugs or effective drug combinations, are therefore eagerly awaited. Tinostamustine is an alkylating and histone deacetylase inhibiting molecule with great potential in cancer treatment. Thus, the aim of this study was to investigate its effects on glioma cells. In this context, tinostamustine was evaluated in monotherapy and as a combination therapy, with either celecoxib or temozolomide; additionally, the results were compared to the golden glioma chemotherapy standard—temozolomide. Our experiments, conducted on both temozolomide-sensitive U-87 MG astrocytoma and temozolomide-resistant U-138 MG glioblastoma cells revealed that tinostamustine and its combination with either celecoxib or temozolomide exert dose-dependent cytotoxicity, cause cell cycle arrest, induce oxidative stress-mediated apoptosis of malignant glioma cells, and mitigate their migratory potential. Astrocytoma cells were more susceptible to the tested treatments than glioblastoma cells, and, generally, those dual therapies were superior in anti-glioma efficacy compared to temozolomide. Overall, our study provides evidence that tinostamustine and the combination therapies consisting of tinostamustine and celecoxib or tinostamustine and temozolomide may represent a new approach for the effective treatment of malignant gliomas. Full article

The latest World Health Organization (WHO) classification of central nervous system tumors (WHO2021/5th) has incorporated molecular information into the diagnosis of each brain tumor type including diffuse glioma. Therefore, an artificial intelligence (AI) framework for learning histological patterns and predicting important genetic events would be useful for future studies and applications. Using the concept of multiple-instance learning, we developed an AI framework named GLioma Image-level and Slide-level gene Predictor (GLISP) to predict nine genetic abnormalities in hematoxylin and eosin sections: IDH1/2, ATRX, TP53 mutations, TERT promoter mutations, CDKN2A/B homozygous deletion (CHD), EGFR amplification (EGFRamp), 7 gain/10 loss (7+/10−), 1p/19q co-deletion, and MGMT promoter methylation. GLISP consists of a pair of patch-level GLISP-P and patient-level GLISP-W models, each pair of which is for a genetic prediction task, providing flexibility in clinical utility. In this study, the Cancer Genome Atlas whole-slide images (WSIs) were used to train the model. A total of 108 WSIs from the Tokyo Women’s Medical University were used as the external dataset. In cross-validation, GLISP yielded patch-level/case-level predictions with top performances in IDH1/2 and 1p/19q co-deletion with average areas under the curve (AUCs) of receiver operating characteristics of 0.75/0.79 and 0.73/0.80, respectively. In external validation, the patch-level/case-level AUCs of IDH1/2 and 1p/19q co-deletion detection were 0.76/0.83 and 0.78/0.88, respectively. The accuracy in diagnosing IDH-mutant astrocytoma, oligodendroglioma, and IDH-wild-type glioblastoma was 0.66, surpassing the human pathologist average of 0.62 (0.54–0.67). In conclusion, GLISP is a two-stage AI framework for histology-based prediction of genetic events in adult gliomas, which is helpful in providing essential information for WHO 2021 molecular diagnoses. Full article