Diagnostics, Volume 10, Issue 3 (March 2020) – 53 articles

Glioblastoma (GBM), the most common and aggressive brain tumor, has a very poor outcome and high tumor recurrence rate. The immune system has positive interactions with the central nervous system. Despite many studies investigating immune prognostic factors, there is no effective model to identify predictive biomarkers for GBM. Genomic data and clinical characteristic information of patients with GBM were evaluated by Kaplan–Meier analysis and proportional hazard modeling. Deseq2 software was used for differential expression analysis. Immune-related genes from ImmPort Shared Data and the Cistrome Project were evaluated. The model performance was determined based on the area under the receiver operating characteristic (ROC) curve. CIBERSORT was used to assess the infiltration of immune cells. The results of differential expression analyses showed a significant difference in the expression levels of 2942 genes, comprising 1338 upregulated genes and 1604 downregulated genes (p < 0.05). A population of 24 immune-related genes that predicted GBM patient survival was identified. A risk score model established on the basis of the expressions of the 24 immune-related genes was used to evaluate a favorable outcome of GBM. Further validation using the ROC curve confirmed the model was an independent predictor of GBM (AUC = 0.869). In the GBM microenvironment, eosinophils, macrophages, activated NK cells, and follicular helper T cells were associated with prognostic risk. Our study confirmed the importance of immune-related genes and immune infiltrates in predicting GBM patient prognosis. Full article

Chronic kidney disease (CKD) is characterized by a progressive loss of renal function and a decrease of glomerular filtration rate. Reduced mitochondrial function, coenzyme Q₁₀ (CoQ₁₀), and increased oxidative stress in patients with CKD contribute to the disease progression. We tested whether CoQ₁₀ levels, oxidative stress and platelet mitochondrial bioenergetic function differ between groups of CKD patients. Methods: Twenty-seven CKD patients were enrolled in this trial, 17 patients had arterial hypertension (AH) and 10 patients had arterial hypertension and diabetes mellitus (AH and DM). The control group consisted of 12 volunteers. A high-resolution respirometry (HRR) method was used for the analysis of mitochondrial bioenergetics in platelets, and an HPLC method with UV detection was used for CoQ₁₀ determination in platelets, blood, and plasma. Oxidative stress was determined as thiobarbituric acid reactive substances (TBARS). Results: Platelets mitochondrial respiration showed slight, not significant differences between the groups of CKD patients and control subjects. The oxygen consumption by intact platelets positively correlated with the concentration of CoQ₁₀ in the platelets of CKD patients. Conclusion: A decreased concentration of CoQ₁₀ and oxidative stress could contribute to the progression of renal dysfunction in CKD patients. The parameters of platelet respiration assessed by high-resolution respirometry can be used only as a weak biological marker for mitochondrial diagnosis and therapy monitoring in CKD patients. Full article

Acute subdural hematoma (aSDH) is a common pathology encountered after head trauma. Only a minority of aSDHs have an arterial source. In this article, we report a case of aSDH originating from a traumatic pseudoaneurysm of the distal segment of posterior cerebral artery (PCA), diagnosed several days after the initial minor trauma and successfully treated with endovascular coiling. This case emphasizes the importance of searching for vascular pathology when the localization, severity or relapsing course of the intracranial hemorrhage does not fully correspond to the severity of initial trauma and when the bleeding has a delayed onset. Characteristics, diagnostics and treatment possibilities of traumatic cerebral aneurysms, an important cause of arterial aSDH, are described in the article. Full article

Introduction: Traumatic brain injuries (TBI) are a real social problem, with an upward trend worldwide. The most frequent consequence of a traumatic brain injury is extra-axial hemorrhage, i.e., an acute subdural (SDH) and epidural hematoma (EDH). Most of the factors affecting the prognosis have been analyzed on a wide group of traumatic brain injuries. Nonetheless, there are few studies analyzing factors influencing the prognosis regarding patients undergoing surgery due to acute subdural and epidural hematoma. The aim of this study was to identify the factors which have the strongest prognostic value in relation to the 6-month outcome of the patients undergoing surgery for SDH and EDH. Patients and methods: The study included a group of 128 patients with isolated craniocerebral injuries. Twenty eight patients were operated upon due to EDH, and a group of 100 patients were operated upon due to SDH. The following factors from the groups were analyzed: demographic data, physiological factors, laboratory factors, computed tomography scan characteristics, and time between the trauma and the surgery. All of these factors were correlated in a multivariate analysis with the six-month outcome in the Glasgow outcome scale. Results: The factors with the strongest prognostic value are GCS score, respiration rate, saturation, glycaemia and systolic blood pressure. Conclusion: Initial GCS score, respiratory rate, saturation, glycaemia and systolic blood pressure were the factors with the strongest prognostic value. Full article

Background: A test to narrow down patients who require esophagogastroduodenoscopy (EGD) with a high probability of having gastroesophageal varices (GEV) and a high-risk of liver-related events is an unmet need. Methods: The measurement of serum fibrosis markers and EGD was performed in 166 consecutive chronic hepatitis C patients. The correlation between the grades of GEV and fibrosis markers and the subsequent occurrence of liver-related and fibrosis markers were examined. Results: Wisteria floribunda agglutinin-positive human Mac-2 binding protein (WFA⁺–M2BP) levels increased according to the grade of GEV (3.4 (0.2–18.6) for no GEV, 7.9 (1.8–20.0) for small GEV, and 11.4 (4.0–20.0) for large GEV; p < 0.001). The diagnostic accuracy of the WFA⁺–M2BP was superior compared to other serum fibrosis markers, and WFA⁺–M2BP was an independent predictor of GEV in the multivariate analysis. Furthermore, the cumulative incidence of liver-related events at one year was 2.3% in patients with WFA⁺–M2BP levels ≤ 7.0 and 37.5% in patients with WFA⁺–M2BP levels > 7.0 (p < 0.001). WFA⁺–M2BP > 7.0 was a significant predictive factor for liver-related events (Hazard ratio 6.7, p = 0.004) independent of Child–Pughclass. Conclusions: WFA⁺–M2BP could be used to estimate the presence and grade of GEV and is linked to liver-related events in chronic hepatitis C patients. Full article

The mucopolysaccharidoses (MPSs) include 11 different conditions caused by specific enzyme deficiencies in the degradation pathway of glycosaminoglycans (GAGs). Although most MPS types present increased levels of GAGs in tissues, including blood and urine, diagnosis is challenging as specific enzyme assays are needed for the correct diagnosis. Enzyme assays are usually performed in blood, with some samples (as leukocytes) providing a final diagnosis, while others (such as dried blood spots) still being considered as screening methods. The identification of variants in the specific genes that encode each MPS-related enzyme is helpful for diagnosis confirmation (when needed), carrier detection, genetic counseling, prenatal diagnosis (preferably in combination with enzyme assays) and phenotype prediction. Although the usual diagnostic flow in high-risk patients starts with the measurement of urinary GAGs, it continues with specific enzyme assays and is completed with mutation identification; there is a growing trend to have genotype-based investigations performed at the beginning of the investigation. In such cases, confirmation of pathogenicity of the variants identified should be confirmed by measurement of enzyme activity and/or identification and/or quantification of GAG species. As there is a growing number of countries performing newborn screening for MPS diseases, the investigation of a low enzyme activity by the measurement of GAG species concentration and identification of gene mutations in the same DBS sample is recommended before the suspicion of MPS is taken to the family. With specific therapies already available for most MPS patients, and with clinical trials in progress for many conditions, the specific diagnosis of MPS as early as possible is becoming increasingly necessary. In this review, we describe traditional and the most up to date diagnostic methods for mucopolysaccharidoses. Full article

This study aimed to find potential diagnostic markers for osteoarthritis (OA) and analyze the role of immune cells infiltration in this pathology. We used OA datasets from the Gene Expression Omnibus database. First, R software was used to identify differentially expressed genes (DEGs) and perform functional correlation analysis. Then least absolute shrinkage and selection operator (LASSO) logistic regression and support vector machine-recursive feature elimination algorithms were used to screen and verify the diagnostic markers of OA. Finally, CIBERSORT was used to evaluate the infiltration of immune cells in OA tissues, and the correlation between diagnostic markers and infiltrating immune cells was analyzed. A total of 458 DEGs were screened in this study. GRB10 and E2F3 (AUC = 0.962) were identified as diagnostic markers of OA. Immune cell infiltration analysis found that resting mast cells, T regulatory cells, CD4 memory resting T cells, activated NK cells, and eosinophils may be involved in the OA process. In addition, GRB10 was correlated with NK resting cells, naive CD4 + T cells, and M1 macrophages, while E2F3 was correlated with resting mast cells. In conclusion, GRB10 and E2F3 can be used as diagnostic markers of osteoarthritis, and immune cell infiltration plays an important role in the occurrence and progression of OA. Full article

Measurement of gastric emptying is of clinical value for a range of conditions. Gamma scintigraphy (GS) has an established role, but the use of magnetic resonance imaging (MRI) has recently increased. Previous comparison studies between MRI and GS showed good correlation, but were performed on separate study days. In this study, the modalities were alternated rapidly allowing direct comparison with no intra-individual variability confounds. Twelve healthy participants consumed 400 g of Technetium-99m (^99mTc)-labelled soup test meal (204 kcal) and were imaged at intervals for 150 min, alternating between MRI and GS. The time to empty half of the stomach contents (T_1/2) and retention rate (RR) were calculated and data correlated. The average T_1/2 was similar for MRI (44 ± 6 min) and GS (35 ± 4 min) with a moderate but significant difference between the two modalities (p < 0.004). The individual T_1/2 values were measured, and MRI and GS showed a good positive correlation (r = 0.95, p < 0.0001), as well as all the RRs at each time point up to 120 min. Gastric emptying was measured for the first time by MRI and GS on the same day. This may help with translating the use of this simple meal, known to elicit reliable, physiological, and pathological gastrointestinal motor, peptide, and appetite responses. Full article

A growing number of research papers regarding Spanish-speaking dancers justifies the need for an adapted Spanish version of the Dance Functional Outcome Survey (DFOS). The objective of this study was to cross-culturally adapt and validate the DFOS for Spanish-speaking dancers. A sample of 127 healthy and injured professional and pre-professional dancers were recruited. Test-retest reliability of DFOS-Sp was examined using intraclass correlation coefficients. Construct validity compared DFOS-Sp to the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) using Pearson correlations. Principal component analysis identified factors and internal-item consistency. Sensitivity was evaluated by generating receiver operating characteristic and area under the curve analyses. A subgroup of 51 injured dancers were followed across three time-points to examine responsiveness using repeated measures analysis of variance. Injured scores were analyzed for floor and ceiling effects. The DFOS-Sp showed high test-retest reliability (ICC2,1 ≥ 0.92). DFOS-Sp scores had moderate construct validity compared with SF-36 physical component summary scores (r ≥ 0.56). Principal component analysis (PCA) supported uni-dimensionality explaining 58% of the variance with high internal consistency (α = 0.91).Area under the curve (AUC) sensitivity values were excellent (AUC ≥ 0.82). There were significant differences across time (p < 0.001), demonstrating responsiveness to change, with no floor or ceiling effects. The DFOS-Sp demonstrated acceptable test-retest reliability and validity in Spanish-speaking dancers, with comparable psychometric performance to the English-language version. Full article

Multiple primary malignant tumors are defined by the development of more than one malignancy in a single patient. These can be subdivided into synchronous or metachronous malignant tumors, depending on their time of appearance, relative to the first malignancy. Triple primary malignancies are a relatively rare occurrence in day-to-day practice and triple synchronous or metachronous carcinomas involving a thyroid localization are even less common. In this case series, we report our 20-year experience in diagnosing and managing five patients with triple malignant tumors with thyroid involvement. Full article

Background: Intensive and prolonged exercise leads to a rise of troponin concentration in blood. The mechanism responsible for troponin release during exercise remains ill-defined. The study aim was to search for risk factors of troponin increase after a prolonged endurance competition. Methods: The study included a group of 18 amateurs, healthy volunteers (median age 41.5 years, interquartile range – IQR 36–53 years, 83% male) who participated in a 100 km running ultra-marathon. Information on demographic characteristics, pre- and post-race heart rate, blood pressure, body composition and glucose, lactate (L), troponin T (hs-TnT) and C reactive protein (hs-CRP) concentration were obtained. Additionally, data on L and glucose levels every 9.2 km and fluid/food intakes during the race were collected. Results: There was a significant hs-TnT increase after the race exceeding upper reference values in 66% of runners (from 5 IQR 3–7 ng/L to 14 IQR 12–26 ng/L, p < 0.0001). None of the baseline parameters predicted a post-race hs-TnT increase. The only factors, correlating with changes of hs-TnT were mean L concentration during the race (rho = 0.52, p = 0.03) and change of hs-CRP concentration (rho = 0.59, p = 0.01). Conclusions: Participation in a 100 km ultra-marathon leads to a modest, but significant hs-TnT increase in the majority of runners. Among analysed parameters only mean lactate concentration during the race and change in hs-CRP correlated with troponin change. Full article

Background: Primary dysmenorrhea (PD) is one of the most common gynecological disorders in women of reproductive age that may affect quality of life. It is believed that the underlying cause of PD may be the excessive production of prostaglandins (PGs), however, between 20%–25% of women with PD did not respond to pharmacological treatments, showing that nociceptive mechanisms underlying PD are still not understood. The purpose of this study was to measure and compare, through the use of ultrasound imaging, the thickness at rest of the abdominal wall, as well as the interrecti distance (IRD), in women with and without PD. Methods: A cross-sectional study has been performed using ultrasound imaging (USI) to measure the resting thickness of the external oblique (EO), internal oblique (IO), transversus abdominis (TrAb), rectus abdominis (RA), as well as the IRD in a sample of 39 women, 19 with PD and 20 without PD (median ± IR age: 20 ± 4 and 22.5 ± 7 years, respectively). Results: Findings of muscular thickness did not reveal statically significant differences (p < 0.05) in EO, IO, TrAb, RA, and the IRD between the PD group and control group. Conclusions: These findings suggest that the thickness of the abdominal wall is not associated with PD. Full article

The role of dynamic contrast-enhanced-MRI (DCE-MRI) for Prostate Imaging-Reporting and Data System (PI-RADS) scoring is a controversial topic. In this retrospective study, we aimed to measure the added value of DCE-MRI in combination with T2-weighted (T2W) and diffusion-weighted imaging (DWI) using PI-RADS v2.1, in terms of reproducibility and diagnostic accuracy, for detection of prostate cancer (PCa) and clinically significant PCa (CS-PCa, for Gleason Score ≥ 7). 117 lesions in 111 patients were identified as suspicion by multiparametric MRI (mpMRI) and addressed for biopsy. Three experienced readers independently assessed PI-RADS score, first using biparametric MRI (bpMRI, including DWI and T2W), and then multiparametric MRI (also including DCE). The inter-rater and inter-method agreement (bpMRI- vs. mpMRI-based scores) were assessed by Cohen’s kappa (κ). Receiver operating characteristics (ROC) analysis was performed to evaluate the diagnostic accuracy for PCa and CS-PCa detection among the two scores. Inter-rater agreement was excellent for the three pairs of readers (κ ≥ 0.83), while the inter-method agreement was good (κ ≥ 0.73). Areas under the ROC curve (AUC) showed similar high-values (0.8 ≤ AUC ≤ 0.85). The reproducibility of PI-RADS v2.1 scoring was comparable and high among readers, without relevant differences, depending on the MRI protocol used. The inclusion of DCE did not influence the diagnostic accuracy. Full article

This work aimed to identify and compare the bacterial patterns present in endometriotic lesions, eutopic endometrium and vaginal fluid from endometriosis patients with those found in the vaginal fluid and eutopic endometrium of control patients. Vaginal fluid, eutopic endometrium and endometriotic lesions were collected. DNA was extracted and the samples were analyzed to identify microbiome by high-throughput DNA sequencing of the 16S rRNA marker gene. Amplicon sequencing from vaginal fluid, eutopic endometrium and endometriotic lesion resulted in similar profiles of microorganisms, composed most abundantly by the genus Lactobacillus, Gardnerella, Streptococcus and Prevotella. No significant differences were found in the diversity analysis of microbiome profiles between control and endometriotic patients; however deep endometriotic lesions seems to present different bacterial composition, less predominant of Lactobacillus and with more abundant Alishewanella, Enterococcus and Pseudomonas. Full article

Major Depression Disease has been increasing in the last few years, affecting around 7 percent of the world population, but nowadays techniques to diagnose it are outdated and inefficient. Motor activity data in the last decade is presented as a better way to diagnose, treat and monitor patients suffering from this illness, this is achieved through the use of machine learning algorithms. Disturbances in the circadian rhythm of mental illness patients increase the effectiveness of the data mining process. In this paper, a comparison of motor activity data from the night, day and full day is carried out through a data mining process using the Random Forest classifier to identified depressive and non-depressive episodes. Data from Depressjon dataset is split into three different subsets and 24 features in time and frequency domain are extracted to select the best model to be used in the classification of depression episodes. The results showed that the best dataset and model to realize the classification of depressive episodes is the night motor activity data with 99.37% of sensitivity and 99.91% of specificity. Full article

Mucopolysaccharidosis type I (MPS I) is caused by the deficiency of α-l-iduronidase, leading to the storage of dermatan and heparan sulfate. There is a broad phenotypical spectrum with the presence or absence of neurological impairment. The classical form is known as Hurler syndrome, the intermediate form as Hurler–Scheie, and the most attenuated form is known as Scheie syndrome. Phenotype seems to be largely influenced by genotype. Patients usually develop several somatic symptoms such as abdominal hernias, extensive dermal melanocytosis, thoracolumbar kyphosis odontoid dysplasia, arthropathy, coxa valga and genu valgum, coarse facial features, respiratory and cardiac impairment. The diagnosis is based on the quantification of α-l-iduronidase coupled with glycosaminoglycan analysis and gene sequencing. Guidelines for treatment recommend hematopoietic stem cell transplantation for young Hurler patients (usually at less than 30 months of age). Intravenous enzyme replacement is approved and is the standard of care for attenuated—Hurler–Scheie and Scheie—forms (without cognitive impairment) and for the late-diagnosed severe—Hurler—cases. Intrathecal enzyme replacement therapy is under evaluation, but it seems to be safe and effective. Other therapeutic approaches such as gene therapy, gene editing, stop codon read through, and therapy with small molecules are under development. Newborn screening is now allowing the early identification of MPS I patients, who can then be treated within their first days of life, potentially leading to a dramatic change in the disease’s progression. Supportive care is very important to improve quality of life and might include several surgeries throughout the life course. Full article

Ankle/foot pain is a common complaint encountered in clinical practice. Currently, due to the complex anatomy, the diagnosis and management of the underlying musculoskeletal disorders are extremely challenging. Nowadays, high-resolution ultrasound has emerged as the first-line tool to evaluate musculoskeletal disorders. There have been several existing protocols describing the fundamental sonoanatomy of ankle/foot joints. However, there are certain anatomic structures (e.g., Lisfranc ligament complex or Baxter nerve) which are also clinically important. As they are rarely elaborated in the available literature, a comprehensive review is necessary. In this regard, the present article aims to brief the regional anatomy, illustrate the scanning techniques, and emphasize the clinical relevance of the ankle/foot region. Full article

Patients with type 2 diabetes mellitus (T2DM) are at increased risk of non-alcoholic fatty liver disease (NAFLD) and might eventually progress to advanced fibrosis, cirrhosis and hepatocellular carcinoma (HCC). Recommendations on whether to screen for NAFLD in diabetic patients remains conflicted between major guidelines. Transient elastography using FibroScan with CAP (controlled attenuation parameter) can assess both liver steatosis and fibrosis simultaneously. This paper took a new look at the prevalence of NAFLD and the severity of fibrosis among T2DM patients in Vietnam. The study was conducted using a cross-sectional design in T2DM adults who attended Dai Phuoc Ho Chi Minh Polyclinic and Polyclinic of Pham Ngoc Thach University of Medicine. Liver steatosis and fibrosis was assessed by FibroScan. NAFLD was diagnosed if CAP > 233 dB/m (steatosis > 5%). Data were analyzed using STATA 12 software program. We found that a total of 307 type 2 diabetic patients qualified for the study’s criteria. The prevalence of NAFLD in T2DM patients based on FibroScan was 73.3%. Rates of mild, moderate and severe steatosis were 20.5%, 21.8% and 30.9%, respectively. The prevalence of significant fibrosis (≥ F2), advanced fibrosis (≥ F3) and cirrhosis (F4) was 13.0%, 5.9% and 3.6%, respectively. On multivariate analysis, aspartate aminotransferase (AST) (OR: 1.067; 95% CI: 1.017–1.119; p = 0.008) and platelet levels (OR: 0.985; 95% CI: 0.972–0.999; p = 0.034) were independent of risk factors of advanced fibrosis. Thus, our study supports screening for NAFLD and for evaluating the severity of liver fibrosis in T2DM patients. Full article

Health economics is a discipline of economics applied to health care. One method used in health economics is decision tree modelling, which extrapolates the cost and effectiveness of competing interventions over time. Such decision tree models are the basis of reimbursement decisions in countries using health technology assessment for decision making. In many instances, these competing interventions are diagnostic technologies. Despite a wealth of excellent resources describing the decision analysis of diagnostics, two critical errors persist: not including diagnostic test accuracy in the structure of decision trees and treating sequential diagnostics as independent. These errors have consequences for the accuracy of model results, and thereby impact on decision making. This paper sets out to overcome these errors using color to link fundamental epidemiological calculations to decision tree models in a visually and intuitively appealing pictorial format. The paper is a must-read for modelers developing decision trees in the area of diagnostics for the first time and decision makers reviewing diagnostic reimbursement models. Full article

Follicular lymphoma, the second most common non-Hodgkin lymphoma (NHL), primarily affects adults and shows an indolent clinical course. Rare cases of follicular lymphoma transform to a high-grade B-cell lymphoma with MYC and BCL2 rearrangements or “double-hit lymphoma”. Transformation to a “double-hit lymphoma” portends a worse prognosis and requires aggressive treatment. We report a comprehensive clinical, pathologic and radiographic review of a patient with previously undiagnosed low-grade follicular lymphoma that transformed into a “double-hit lymphoma”. The patient presented with a large heterogeneous mass 16 x 19 cm involving pancreatic head and neck and a mildly enlarged inguinal lymph node. Positron emission tomography (PET) study demonstrated Fluorodeoxyglucose (¹⁸F) (FDG)-avid peripancreatic mass. Tissue biopsy demonstrated a high-grade B-cell lymphoma with rearrangements t(14;18) and MYC, leading to the diagnosis of high-grade B-cell lymphoma with MYC and BCL2 rearrangements. Excisional biopsy of an inguinal lymph node demonstrated low-grade follicular lymphoma. Clonality studies demonstrated the same immunoglobulin clone V7-4 in inguinal lymph node and peripancreatic mass. Therefore, diagnosis of a high-grade B-cell lymphoma with MYC and BCL2 rearrangements that transformed from a low-grade follicular lymphoma was rendered. It is ultimately important to establish a tissue-based diagnosis at the different sites that are involved with lymphoma. Patient proceeded with the aggressive treatment with dose-adjusted etoposide, prednisone, vincristine, cyclophosphamide, doxorubicin and rituximab (EPOCH-R) treatment. Full article

This paper reports a mechanism for corticospinal tract injury in a patient with hemiplegia following traumatic brain injury (TBI) based on diffusion tensor tractography (DTT) finding. A 73-year-old male with TBI resulting from a fall, without medical history, was diagnosed as having left convexity epidural hematoma (EDH). He underwent craniotomy and suffered motor weakness on the right side of the body. Two weeks after onset, he was transferred to a rehabilitation department with an alerted level of consciousness. Four weeks after onset, his motor functions were grade 1 by the Medical Research Council’s (MRC) standards in the right-side limbs and grade 4 in the left-side limbs. The result of DTT using the different regions of interest (ROIs) showed that most of the right corticospinal tract (CST) did not reach the cerebral cortex around where the EDH was located, and when the ROI was placed on upper pons, a disconnection of the CST was shown and a connection of the CST in ROI with the middle pons appeared. However, the right CST was connected to the cerebral cortex below the pons regardless of ROI. This study is the first report to use DTT to detect that the discontinuation of the left CST in the cerebral cortex and injury lesions below the lower pons and between the upper and lower pons are responsible for motor weakness in a patient. Full article

Mucopolysaccharidoses (MPSs) are a group of lysosomal storage disorders associated with impaired glycosaminoglycans (GAGs) catabolism. In MPS I, II, III, and VII, heparan sulfate (HS) cannot be degraded because of the lack of sufficient activity of the respective enzymes, and its accumulation in the brain causes neurological symptoms. Globotriaosylsphingosine (LysoGb3), the deacylated form of globotriaosylceramide (Gb3), is described as a highly sensitive biomarker for another lysosomal storage disease—Fabry disease. The connection between MPSs and LysoGb3 has not yet been established. This study included 36—MPS I, 15—MPS II, 25—MPS III, 26—MPS IV, and 14—MPS VI patients who were diagnosed by biochemical and molecular methods and a control group of 250 males and 250 females. The concentration of lysosphingolipids (LysoSLs) was measured in dried blood spots by high pressure liquid chromatography—tandem mass spectrometry. We have demonstrated that LysoGb3 concentration was significantly elevated (p < 0.0001) in untreated MPS I (3.07 + 1.55 ng/mL), MPS II (5.24 + 2.13 ng/mL), and MPS III (6.82 + 3.69 ng/mL) patients, compared to the control group (0.87 + 0.55 ng/mL). LysoGb3 level was normal in MPS VI and MPS IVA (1.26 + 0.39 and 0.99 + 0.38 ng/mL, respectively). Activity of α-galactosidase A (α-Gal A), an enzyme deficient in Fabry disease, was not, however, inhibited by heparan sulfate in vitro, indicating that an increase of LysoGb3 level in MPS I, MPS II, and MPS III is an indirect effect of stored MPSs rather than a direct result of impairment of degradation of this compound by HS. Our findings indicate some association of elevated LysoGb3 concentration with the neuronopathic forms of MPSs. The pathological mechanism of which is still to be studied. Full article

The recent eighth tumor-node-metastasis (TMN) staging system classifies renal cell carcinoma (RCC) with perirenal fat invasion (PFI), renal sinus fat invasion (SFI), or renal vein invasion (RVI) as stage pT3a. However, limited data are available on whether these sites have similar prognostic value or recurrence rate. We investigated the recurrence rate based on tumor size, pathological invasion sites including urinary collecting system invasion (UCSI), and clinically detected renal vein thrombus (cd-RVT) with pT3aN0M0 RCC. We retrospectively reviewed 91 patients with pT3aN0M0 RCC who underwent surgical treatment. Patients with tumor size > 7 cm, UCSI, three invasive sites (PFI + SFI + RVI), and cd-RVT showed a significant correlation with high recurrence rates (hazard ration (HR) 2.98, p = 0.013; HR 8.86, p < 0.0001; HR 14.28, p = 0.0008; and HR 4.08, p = 0.0074, respectively). In the multivariate analysis, tumor size of >7 cm, the presence of UCSI, and cd-RVT were the independent predictors of recurrence (HR 3.39, p = 0.043, HR 7.31, p = 0.01, HR 5.06, p = 0.018, respectively). In pT3a RCC, tumor size (7 cm cut-off), UCSI, and cd-RVT may help to provide an early diagnosis of recurrence. Full article

Detection of the Janus Kinase-2 (JAK2) V617F mutation is a diagnostic criterion for myeloproliferative neoplasms, and high levels of mutant alleles are associated with worse outcomes. This mutation is usually tested on blood DNA by allele-specific qPCR (AS-qPCR) and measured using absolute quantification. However, some automated DNA extractions co-extracts of PCR inhibitors from blood and qPCR absolute quantification need increased efforts in order to maintain standard curves. JAK2 V617F can also be detected in serum using droplet digital PCR (ddPCR), a specimen with less inhibitors and favorable to automated extractions, but ddPCR instruments are not wide available as qPCR thermocyclers. Here, we evaluate whether JAK2 V617F could be accurately quantified by AS-qPCR using the 2^-∆∆Cq method on blood DNA and validate the assay using gold-standard molecular diagnostic protocols. Next, we apply the validated method to assess if the mutation could be reliably detected/quantified in serum. JAK2 V617F could be quantified by AS-qPCR using the 2^-∆∆Cq method—the assay was highly accurate (bias of 1.91%) compared to a commercial kit, highly precise (total CV% of 0.40%, 1.92%, 11.12% for samples with 93%, 54%, and 2.5% of mutant allele), highly sensitive (limit of detection of 0.15%), and demonstrated a linear detection response from 1.1% to 99.9%. Serum presented a higher mutant allele burden compared to the paired whole blood (mean of 4%), which allows for an increased JAK2 mutant detection rate and favors increased JAK2 V617F high-throughput analysis. Full article

Current diagnostic methods of acute kidney injury (AKI) have limited sensitivity and specificity. Tissue injury has been linked to an increase in the concentrations of extracellular DNA (ecDNA) in plasma. A rapid turnover of ecDNA in the circulation makes it a potential marker with high sensitivity. This study aimed to analyze the concentration of ecDNA in plasma in animal models of AKI. Three different fractions of ecDNA were measured—total ecDNA was assessed fluorometrically, while nuclear ecDNA (ncDNA) and mitochondrial DNA (mtDNA) were analyzed using quantitative real-time PCR. AKI was induced using four different murine models of AKI-bilateral ureteral obstruction (BUO), glycerol-induced AKI (GLY), ischemia–reperfusion injury (IRI) and bilateral nephrectomy (BNx). Total ecDNA was significantly higher in BUO (p < 0.05) and GLY (p < 0.05) compared to the respective control groups. ncDNA was significantly higher in BUO (p < 0.05) compared to SHAM. No significant differences in the concentrations of mtDNA were found between the groups. The plasma concentrations of different fractions of ecDNA are dependent on the mechanism of induction of AKI and warrant further investigation as potential surrogate markers of AKI. Full article

Background: The AR-V7 splice variant is a cause of castration-resistant prostate cancer (CRPC). However, testing for the presence of AR-V7 by real-time polymerase chain reaction (RT-PCR) shows AR-V7 positivity in healthy individuals. We hypothesized that the positivity reflects contamination by hematopoietic cells. We tried a novel circulating tumor cell (CTC) enrichment instrument, using Celsee, to clear hematopoietic cells. Methods: We tested whole blood or Celsee-enriched samples for AR-V7 by RT-PCR, and included samples from 41 CRPC patients undergoing sequential therapy. We evaluated the associations between AR-V7 status and clinical factors. We evaluated factors affecting AR-V7 positivity. Results: AR-V7 positivity was lower in Celsee-enriched than in whole blood specimens. AR-V7 and clinical factors did not predict the therapy effectiveness. We found no significant differences in the effectiveness of enzalutamide/abiraterone (Enz/Abi) upon AR-V7 evaluation. All AR-V7 positive patients had resistance to Enz/Abi. Docetaxel (DTX), cabazitaxel (CBZ), and Radium223 treatment showed no significant difference in the treatment effectiveness, regardless of AR-V7 presence. AR-V7 was more frequently positive than Extent of disease (EOD) 2 in cases with bone metastases. Conclusion: Celsee CTC enrichment suppresses AR-V7 false positivity. All AR-V7 positive patients presented resistance to Enz/Abi. DTX, CBZ, and Radium223 were effective and remain treatment options. AR-V7 positivity should progressively appear in patients with advanced bone metastases. Full article

Malignancies of extragenital origin very rarely metastasize to the uterine body. Endometrium-limited metastases may pose diagnostic challenges in endometrial curettage specimens as they may be misdiagnosed as primary endometrial tumors. We investigated the clinicopathological characteristics of seven cases with endometrial-limited metastases from carcinomas of the nasopharynx (n = 1), breast (n = 2), colon (n = 2), stomach (n = 1), and appendix (n = 1). The patients’ ages ranged from 36 to 71 (mean: 55.4) years. None of the patients had a remarkable gynecological history, and the presenting sign in all cases was abnormal uterine bleeding. Although myometrial involvement was absent, multiple metastases were already present in extrauterine locations such as the lung, liver, bone, abdominopelvic peritoneum, and omentum. All patients underwent ultrasonographic examination prior to endometrial curettage. The histologies of the endometrial metastases identified from the curettage specimens were identical to those of the corresponding primary tumors. Ancillary tests including immunostaining and Epstein–Barr virus-encoded RNA in situ hybridization confirmed the extragenital origin. Endometrium-limited metastases from extragenital malignancies are extremely rare. They present with abnormal vaginal bleeding and mimic endometrial carcinomas of endometrioid or poorly differentiated types. Since their clinical presentations and histological features are similar to those of primary endometrial tumors, pathologists should consider the possibility of metastases while evaluating endometrial curettage specimens obtained from patients with a history of extragenital malignancies. Full article

Thyroid hormones are important factors that regulate metabolism and cell differentiation throughout the human body. A complication of thyroid pathology is represented by an alteration of the bone metabolism which can lead to osteoporosis and fragility fractures, known to have a high mortality rate. Although there is a consensus on the negative impact of hyperthyroidism on bone metabolism, when referring to hypothyroidism, subclinical hypothyroidism, or subclinical hyperthyroidism, there is no general agreement. The aim of our review was to update clinicians and researchers about the current data regarding the bone health in hypothyroidism, subclinical hypothyroidism, and subclinical hyperthyroidism patients. Thyroid disorders have an important impact on bone metabolism and fracture risk, such that hyperthyroidism, hypothyroidism, and subclinical hyperthyroidism are associated with a decreased bone mineral density (BMD) and increased risk of fracture. Subclinical hypothyroidism, on the other hand, is not associated with osteoporosis or fragility fractures, and subclinical hyperthyroidism treatment with radioiodine could improve bone health. Full article

This study aims to investigate the ability of ultrasound strain elastography as an adjunct to predict malignancy in soft tissue tumors suspect of sarcoma or metastasis in a tertiary reference center for sarcoma. A total of 137 patients were included prospectively. Patients were referred on the basis of clinical or radiological suspicion of malignant soft tissue tumor. All patients had previously undergone diagnostic imaging (MRI, CT or PET-CT). After recording strain elastography cine loops, ultrasound guided biopsy was performed. Three investigators, who were blinded to final diagnosis, reviewed all elastograms retrospectively. For each elastogram, a qualitative, visual 5-point score was decided in consensus and a strain ratio was calculated. Final pathology obtained from biopsy or tumor resection served as gold standard. Eighty-one tumors were benign, and 56 were malignant. t-tests showed a significant difference in mean visual score between benign and malignant tumors. There was no significant difference in mean strain ratio between the two groups. Strain elastography may be a valuable adjunct to conventional B-mode ultrasound, perhaps primarily in primary care, when considering whether to refer to a sarcoma center or to biopsy, although biopsies cannot reliably be ruled out based on the current data. Full article