Metabolic-associated fatty liver voxel-based quantification on CT images using a contrast adapted automatic tool.

BACKGROUND & AIMS: Metabolic-dysfunction associated fatty liver disease (MAFLD) is highly prevalent and can lead to liver complications and comorbidities, with non-invasive tests such as vibration-controlled transient elastography (VCTE) and invasive liver biopsies being used for diagnosis The aim of the present study was to develop a new fully automatized method for quantifying the percentage of fat in the liver based on a voxel analysis on computed tomography (CT) images to solve previously unconcluded diagnostic deficiencies either in contrast (CE) or non-contrast enhanced (NCE) assessments. METHODS: Liver and spleen were segmented using nn-UNet on CE- and NCE-CT images. Radiodensity values were obtained for both organs for defining the key benchmarks for fatty liver assessment: liver mean, liver-to-spleen ratio, liver-spleen difference, and their average. VCTE was used for validation. A classification task method was developed for detection of suitable patients to fulfill maximum reproducibility across cohorts and highlight subjects with other potential radiodensity-related diseases. RESULTS: Best accuracy was attained using the average of all proposed benchmarks being the liver-to-spleen ratio highly useful for CE and the liver-to-spleen difference for NCE. The proposed whole-organ automatic segmentation displayed superior potential when compared to the typically used manual region-of-interest drawing as it allows to accurately obtain the percent of fat in liver, among other improvements. Atypical patients were successfully stratified through a function based on biochemical data. CONCLUSIONS: The developed method tackles the current drawbacks including biopsy invasiveness, and CT-related weaknesses such as lack of automaticity, dependency on contrast agent, no quantification of the percentage of fat in liver, and limited information on region-to-organ affectation. We propose this tool as an alternative for individualized MAFLD evaluation by an early detection of abnormal CT patterns based in radiodensity whilst abording detection of non-suitable patients to avoid unnecessary exposure to CT radiation. Furthermore, this work presents a surrogate aid for assessing fatty liver at a primary assessment of MAFLD using elastography data.

Comparison of Extracellular Vesicle Isolation Methods for miRNA Sequencing.

MicroRNAs (miRNAs) encapsulated in extracellular vesicles (EVs) are potential diagnostic and prognostic biomarkers. However, discrepancies in miRNA patterns and their validation are still frequent due to differences in sample origin, EV isolation, and miRNA sequencing methods. The aim of the present study is to find a reliable EV isolation method for miRNA sequencing, adequate for clinical application. To this aim, two comparative studies were performed in parallel with the same human plasma sample: (i) isolation and characterization of EVs obtained using three procedures: size exclusion chromatography (SEC), iodixanol gradient (GRAD), and its combination (SEC+GRAD) and (ii) evaluation of the yield of miRNA sequences obtained using NextSeq 500 (Illumina) and three miRNA library preparation protocols: NEBNext, NEXTFlex, and SMARTer smRNA-seq. The conclusion of comparison (i) is that recovery of the largest amount of EVs and reproducibility were attained with SEC, but GRAD and SEC+GRAD yielded purer EV preparations. The conclusion of (ii) is that the NEBNext library showed the highest reproducibility in the number of miRNAs recovered and the highest diversity of miRNAs. These results render the combination of GRAD EV isolation and NEBNext library preparation for miRNA retrieval as adequate for clinical applications using plasma samples.

Root-knot nematode assessment: species identification, distribution, and new host records in Portugal.

Considered one of the most devastating plant parasitic nematodes worldwide, Meloidogyne spp. (commonly known as the root-knot nematodes (RKNs)) are obligate sedentary endoparasites that establish in the roots, causing hyperplasia and hypertrophy of surrounding cells, triggering the formation of galls. These galls will affect root development and physiology, leading to substantial yield losses. During 2017-2022, an extensive survey of Meloidogyne species was undertaken in Portugal (mainland and islands). A total of 1,071 samples were collected by the National Plant Protection Organization (DGAV) and private farmers from different regions of the country and were analysed at the Laboratory of Nematology (NemaINIAV). Samples in which the presence of Meloidogyne sp. was detected were used to perform bioassays to obtain females and juveniles for further studies. Since the accurate identification of RKNs is an important aspect of crop management, morphological and biochemical characterisation was performed. The most common morphological features were observed, showing consistency with previous descriptions of the genus. The biochemical identification using the esterase (EST) phenotype revealed the phenotypes of Meloidogyne arenaria, M enterolobi, M. hispanica, M. hapla, M. incognita, M javanica, and M. luci. Meloidogyne incognita and M. javanica were found to be the most prevalent species in the different regions followed by M. arenaria and M. hapla. This is the first distribution report performed in Portugal on RKNs, contributing to the development of management strategies and to updated information on the status of these pests in Europe.

Dasatinib-Induced Chylothorax: A Clinical Laboratory's Perspective.

Dasatinib is a tyrosine kinase inhibitor drug used for chronic myeloid leukaemia (CML) treatment. Chylothorax has been rarely reported as a secondary effect of dasatinib occurring especially in long-term treated patients, although its pathophysiology is not yet fully understood. Laboratory analysis of the pleural effusion is crucial for chylothorax diagnosis. We report a case of a 53-year-old male patient presenting a chylothorax after 14 years of dasatinib therapy where the clinical laboratory was key in the diagnosis.

Insights into Insulin Resistance and Calcification in the Myocardium in Type 2 Diabetes: A Coronary Artery Analysis.

Type 2 diabetes (T2D) is responsible for high incidence of cardiovascular (CV) complications leading to heart failure. Coronary artery region-specific metabolic and structural assessment could provide deeper insight into the extent of the disease and help prevent adverse cardiac events. Therefore, in this study, we aimed at investigating such myocardial dynamics for the first time in insulin-sensitive (mIS) and insulin-resistant (mIR) T2D patients. We targeted global and region-specific variations using insulin sensitivity (IS) and coronary artery calcifications (CACs) as CV risk factor in T2D patients. IS was computed using myocardial segmentation approaches at both baseline and after an hyperglycemic-insulinemic clamp (HEC) on [18F]FDG-PET images using the standardized uptake value (SUV) (ΔSUV = SUVHEC - SUVBASELINE) and calcifications using CT Calcium Scoring. Results suggest that some communicating pathways between response to insulin and calcification are present in the myocardium, whilst differences between coronary arteries were only observed in the mIS cohort. Risk indicators were mostly observed for mIR and highly calcified subjects, which supports previously stated findings that exhibit a distinguished exposure depending on the impairment of response to insulin, while projecting added potential complications due to arterial obstruction. Moreover, a pattern relating calcification and T2D phenotypes was observed suggesting the avoidance of insulin treatment in mIS but its endorsement in mIR subjects. The right coronary artery displayed more ΔSUV, whilst plaque was more present in the circumflex. However, differences between phenotypes, and therefore CV risk, were associated to left descending artery (LAD) translating into higher CACs regarding IR, which could explain why insulin treatment was effective for LAD at the expense of higher likelihood of plaque accumulation. Personalized approaches to assess T2D may lead to more efficient treatments and risk-prevention strategies.

The Northern Root-Knot Nematode Meloidogyne hapla: New Host Records in Portugal.

Root-knot nematodes (RKNs), Meloidogyne spp., are a group of plant-parasitic nematodes (PPNs) of great economic significance worldwide. The northern root-knot nematode, Meloidogyne hapla, is one of the most important species of RKNs occurring in cold regions. In Portugal so far, M. hapla has been found parasitizing potato and fig trees. During surveys carried out in 2019-2022 in fields for horticultural and ornamental production, soil and root samples were collected. Roots were observed under a stereomicroscope to determine the presence of galls. Nematodes were extracted from the soil. Morphological features showed a high similarity and consistency with previous descriptions of the genus. For molecular analysis, total genomic DNA was isolated from single nematodes and used to amplify in multiplex reaction using the species-specific primers JMV1, JMV2 and JMVhapla, and for sequencing of the ITS region with the primers TW81/AB28. Multiplex PCR amplification produced a specific fragment of 440 bp and PCR amplification of the ITS region yielded a single fragment of 550 bp, as expected. The obtained sequences showed a similarity ranging from 99.8% to 100% with the sequences of M. hapla available in the database. The phylogenetic tree revealed that the isolates grouped with M. hapla isolates. From the 690 samples collected, M. hapla was detected in three different hosts (grapevine, eucalyptus and potato) in four districts of mainland Portugal and on Madeira Island. To our knowledge, this is the first report of M. hapla infecting the grapevine and eucalyptus in Portugal.

Identification of Myocardial Insulin Resistance by Using Liver Tests: A Simple Approach for Clinical Practice.

BACKGROUND: We report that myocardial insulin resistance (mIR) occurs in around 60% of patients with type 2 diabetes (T2D) and was associated with higher cardiovascular risk in comparison with patients with insulin-sensitive myocardium (mIS). These two phenotypes (mIR vs. mIS) can only be assessed using time-consuming and expensive methods. The aim of the present study is to search a simple and reliable surrogate to identify both phenotypes. METHODS: Forty-seven patients with T2D underwent myocardial [18F]FDG PET/CT at baseline and after a hyperinsulinemic-euglycemic clamp (HEC) to determine mIR were prospectively recruited. Biochemical assessments were performed before and after the HEC. Baseline hepatic steatosis index and index of hepatic fibrosis (FIB-4) were calculated. Furthermore, liver stiffness measurement was performed using transient elastography. RESULTS: The best model to predict the presence of mIR was the combination of transaminases, protein levels, FIB-4 score and HOMA (AUC = 0.95; sensibility: 0.81; specificity: 0.95). We observed significantly higher levels of fibrosis in patients with mIR than in those with mIS (p = 0.034). In addition, we found that patients with mIR presented a reduced glucose uptake by the liver in comparison with patients with mIS. CONCLUSIONS: The combination of HOMA, protein, transaminases and FIB-4 is a simple and reliable tool for identifying mIR in patients with T2D. This information will be useful to improve the stratification of cardiovascular risk in T2D.

First Detection of Meloidogyne luci (Nematoda: Meloidogynidae) Parasitizing Potato in the Azores, Portugal.

Potato is the third most important crop in the world after rice and wheat, with a great social and economic importance in Portugal as it is grown throughout the country, including the archipelagos of Madeira and the Azores. The tropical root-knot nematode (RKN) Meloidogyne luci is a polyphagous species with many of its host plants having economic importance and the ability to survive in temperate regions, which pose a risk to agricultural production. In 2019, M. luci was detected from soil samples collected from the council of Santo António in Pico Island (Azores). Bioassays were carried out to obtain females, egg masses, and second-stage juveniles to characterize this isolate morphologically, biochemically, and molecularly. The observed morphological features and morphometrics showed high similarity and consistency with previous descriptions. Concerning the biochemical characterization, the esterase (EST) phenotype displayed a pattern with three bands similar to the one previously described for M. luci and distinct from M. ethiopica. Regarding the molecular analysis, an 1800 bp region of the mitochondrial DNA between cytochrome oxidase subunit II (COII) and 16S rRNA genes was analyzed and the phylogenetic tree revealed that the isolate grouped with M. luci isolates (99.17%). This is the first report of M. luci parasitizing potato in the Azores islands, contributing additional information on the distribution of this plant-parasitic nematode.

Expanding the Clinical and Genetic Spectra of Primary Immunodeficiency-Related Disorders With Clinical Exome Sequencing: Expected and Unexpected Findings.

Primary immunodeficiencies (PIDs) refer to a clinically, immunologically, and genetically heterogeneous group of over 350 disorders affecting development or function of the immune system. The increasing use of next-generation sequencing (NGS) technology has greatly facilitated identification of genetic defects in PID patients in daily clinical practice. Several NGS approaches are available, from the unbiased whole exome sequencing (WES) to specific gene panels. Here, we report on a 3-year experience with clinical exome sequencing (CES) for genetic diagnosis of PIDs. We used the TruSight One sequencing panel, which includes 4,813 disease-associated genes, in 61 unrelated patients (pediatric and adults). The analysis was done in 2 steps: first, we focused on a virtual PID panel and then, we expanded the analysis to the remaining genes. A molecular diagnosis was achieved in 19 (31%) patients: 12 (20%) with mutations in genes included in the virtual PID panel and 7 (11%) with mutations in other genes. These latter cases provided interesting and somewhat unexpected findings that expand the clinical and genetic spectra of PID-related disorders, and are useful to consider in the differential diagnosis. We also discuss 5 patients (8%) with incomplete genotypes or variants of uncertain significance. Finally, we address the limitations of CES exemplified by 7 patients (11%) with negative results on CES who were later diagnosed by other approaches (more specific PID panels, WES, and comparative genomic hybridization array). In summary, the genetic diagnosis rate using CES was 31% (including a description of 12 novel mutations), which rose to 42% after including diagnoses achieved by later use of other techniques. The description of patients with mutations in genes not included in the PID classification illustrates the heterogeneity and complexity of PID-related disorders.

Baseline reef health surveys at Bangka Island (North Sulawesi, Indonesia) reveal new threats.

Worldwide coral reef decline appears to be accompanied by an increase in the spread of hard coral diseases. However, whether this is the result of increased direct and indirect human disturbances and/or an increase in natural stresses remains poorly understood. The provision of baseline surveys for monitoring coral health status lays the foundations to assess the effects of any such anthropogenic and/or natural effects on reefs. Therefore, the objectives of this present study were to provide a coral health baseline in a poorly studied area, and to investigate possible correlations between coral health and the level of anthropogenic and natural disturbances. During the survey period, we recorded 20 different types of coral diseases and other compromised health statuses. The most abundant were cases of coral bleaching, followed by skeletal deformations caused by pyrgomatid barnacles, damage caused by fish bites, general pigmentation response and galls caused by cryptochirid crabs. Instances of colonies affected by skeletal eroding bands, and sedimentation damage increased in correlation to the level of bio-chemical disturbance and/or proximity to villages. Moreover, galls caused by cryptochirid crabs appeared more abundant at sites affected by blast fishing and close to a newly opened metal mine. Interestingly, in the investigated area the percentage of corals showing signs of 'common' diseases such as black band disease, brown band disease, white syndrome and skeletal eroding band disease were relatively low. Nevertheless, the relatively high occurrence of less common signs of compromised coral-related reef health, including the aggressive overgrowth by sponges, deserves further investigation. Although diseases appear relatively low at the current time, this area may be at the tipping point and an increase in activities such as mining may irredeemably compromise reef health.

Niveles de PAPP-A y fßhCG en gestantes gemelares monocoriónicas y bicoriónicas espontáneas y sus implicaciones en el cálculo del riesgo del cribado prenatal del primer trimestre / Papp-a and ß - hCG levels in spontaneous monochorionic and dichorionic twin pregnancies and their implications in calculating the prenatal risk in first trimester screening

Objetivo: Evaluar los niveles de distribución de los valores bioquímicos de PAPP-A y fßhCG según su edad gestacional, en gestaciones gemelares monocoriónicas y bicoriónicas concebidas espontáneamente en nuestra población. Material y métodos: Se ha estudiado un grupo de 123 gestantes gemelares. Un subgrupo de 72 gestantes gemelares bicoriónicas, con una edad media de 32,1 años, y otro de 41 gestantes gemelares monocoriónicas, con una edad media de 31,1 años. Resultados: Las gestantes gemelares bicoriónicas presentan valores estadísticamente superiores a las monocoriónicas para los valores de las medias de PAPP-A (MoM) (2,55 versus 1,79; p < 0,001) y de fßhCG (MoM) (2,18 versus 1,70; p < 0,001), y no estadísticamente significativos en las medias de la translucencia nucal (Tn) (MoM) (0,96 versus 0,93; p = 0,3). Se analizan las diferentes distribuciones de las concentraciones de las curvas de normalidad de estos parámetros según edad gestacional y corionicidad. Conclusiones: Es necesario determinar ecográficamente si los gemelos son mono- o bicoriónicos ya que hemos comprobado que en nuestra población presentan curvas de normalidad diferenciadas para PAPP-A y fßhCG, utilizadas para el cálculo del riesgo de aneuploidías en el cribado prenatal de primer trimestre (AU)

Objective: To analyse the distribution of PAPP-A and fß-hCG levels in monochorionic and dichorionic twin pregnancies conceived spontaneously in our population. Materials and methods: One hundred twenty-three twin pregnancies were studied. A group of 72 dichorionic twin pregnancies, with an average age of 32.1 years, and another 32 monochorionic twin pregnancies, with an average age of 31.1 years. Results: Dichorionic twin pregnancies conceived spontaneously show values statistically higher than monochorionic of PAPP-A (MoM) (2.55 versus 1.79, P>.001) and fβHCG (MoM) (2.18 versus 1.70, P>.001). There were no significant differences in nuchal traslucency (NT) between both groups (MoM) (0.96 versus 0.93, P=.3). The distributions of PAPP-A and fßhCG levels according the gestation age and chorionic state have been studied. Conclusions: Ultrasound needs to be performed to determine whether twins are mono- or bichorionics as our population has different distribution curves for PAPP-A and fßhCG, which are used to calculate the first trimester prenatal risk (AU)

Niveles de PAPP-A y fßhCG en gestantes gemelares bicoriónicas concebidas por técnicas de reproducción asistida (FIV-IAH) y sus implicaciones en el cálculo del riesgo del cribado prenatal del primer trimestre / PAPP-A and fBhCG levels in dichorionic twin pregnancies conceveid by assisted reproductive technologies (IVF-AIH) and their implications to calculate the prenatal risk on first trimestry

Objetivo: Evaluar los niveles de proteína plasmática placentaria (PAPP-A) y fracción libre de la βhCG (fßhCG) en gestaciones gemelares bicoriónicas concebidas mediante técnicas de reproducción asistida tipo fertilización in vitro (FIV) o inseminación artificial (IA) en nuestra población. Posterior cálculo de los factores de corrección necesarios a aplicar para el screening de primer trimestre en gestantes gemelares concebidas mediante FIV. Población en estudio: Se han estudiado 171 gestantes gemelares. Un grupo compuesto por 71 gestaciones bicoriónicas concebidas mediante FIV, con una edad media de 36,3 años; un grupo control de 76 gestaciones bicoriónicas concebidas por gestación espontánea, con una edad media de 31,8 años; y un último grupo de 24 gestaciones bicoriónicas concebidas por IA, con una edad media de 32,9 años. Resultados: Las gestantes gemelares bicoriónicas concebidas mediante FIV respecto el grupo control, concebidas espontáneamente, presentan unos niveles estadísticamente inferiores de PAPP-A (2,17 MoM versus 2,41 MoM, p<0,008) y valores superiores no significativos de fßhCG (2,33 MoM versus 2,05 MoM, p=0,144). Las medias de la translucencia nucal (Tn) entre ambos grupos tampoco muestran diferencias significativas (p=0,178). Esta misma tendencia se observa en los valores de PAPPA y fßhCG para las gestaciones concebidas mediante IA, pero con un número menor de gestantes no significativo. Conclusiones: Las mujeres con gestaciones gemelares concebidas por técnicas de reproducción asistida han sido sometidas a tratamientos para la estimulación de la ovulación. Este hecho se ha postulado como una de las causas que podrían explicar la presencia de valores de PAPP-A significativamente inferiores y de fßhCG superiores, aunque no significativos, a las gestantes gemelares espontáneas. Por ello se aconseja la elaboración de curvas propias con gestantes gemelares FIV, o en su defecto, el empleo de factores de corrección para disminuir este efecto. Se comprueba que con la aplicación de estos factores de corrección la tasa de falsos positivos de los gemelares bicoriónicos concebidos por FIV baja de 4,22% a 1,41% en nuestra población, con la consecuente disminución de biopsias de corion que son especialmente complejas en gestaciones gemelares (AU)

Objective: Evaluation of levels of PAPP-A and fßhCG in dichorionic twin pregnancies conceived by Assisted Reproductive Technology, i.e. In Vitro Fertilization (IVF) and Artificicial Insemination (AI) in our population. Calculate the necessary correction factors for first trimester screening in pregnancies conceived by IVF. Study population: One hundred seventyone twin pregnancies were considered within this study. One group was composed by 71 dichorionic twin pregnancies conceived by IVF with an average maternal age of 36.6 years. The control group was composed by 76 dichorionic twin pregnancies conceived spontaneously with an average maternal age of 31.8. A third group was composed by 24 dichorionic twin pregnancies conceived by AI with an average maternal age of 32.9 years. Results: Dichorionic twin pregnancies conceived by IVF present statistically significant lower values for PAPP-A than the control group (2.17 MoM versus 2.41 MoM, p<0.008) and higher values of fßhCG though this difference is not significant (2.33 MoM versus 2.05 MoM, p=0.144). In the other hand, average nuchal translucency (NT) does not show statistical differences between these two groups (p=0.178). Twin pregnancies conceived by AI present these same results tendencies for PAPP-A, fßhCG, and NT, with a lower number of patients not significant. Conclusions: Women with twin pregnancies conceived by Assisted Reproductive Technology (ART) have undergone treatments to induce ovulation. These treatments are proposed as one of the causes that may justify lower PAPP-A values and higher (though not significant) fßhCG values, when twin pregnancies conceived by ART are compared to spontaneously conceived twin pregnancies. Construction of normality curves for twin pregnancies conceived by IVF or calculation of correction factors is advised, in order to decrease this effect in the studied biochemical parameters. Applying these correction factors to our population data, the rate of false positives in first trimester screenings in dichorionic twin pregnancies conceived by IVF is decreased from a 4.22% to a 1.41%; this consequently implies a reduction of chorion biopsies, which are specially complicated in twin pregnancies (AU)