Aging of functional ovaries occurs many years before aging of other organs in the female body. In recent years, a greater number of women continue to postpone their pregnancies to later stages in their lives, raising concerns of the effect of ovarian aging. Mitochondria play an important role in the connection between the aging granulosa cells and oocytes. However, the underlying mechanisms of mitochondrial dysfunction in these cells remain poorly understood. Therefore, we evaluated the molecular mechanism of the aging granulosa cells, including aspects such as accumulation of mitochondrial reactive oxygen species, reduction of mtDNA, imbalance of mitochondrial dynamics, and diminished cell proliferation. Here, we applied bioinformatics approaches, and integrated publicly available resources, to investigate the role of CREB1 gene expression in reproduction. Senescence hallmark enrichment and pathway analysis suggested that the downregulation of bioenergetic-related genes in CREB1. Gene expression analyses showed alterations in genes related to energy metabolism and ROS production in ovary tissue. We also demonstrate that the biogenesis of aging granulosa cells is subject to CREB1 binding to the PRKAA1 and PRKAA2 upstream promoters. In addition, cofactors that regulate biogenesis significantly increase the levels of SIRT1 and PPARGC1A mRNA in the aging granulosa cells. These findings demonstrate that CREB1 elevates an oxidative stress-induced senescence in granulosa cells by reducing the mitochondrial function. Full article

Interest in newborn screening for mucopolysaccharidoses (MPS) is growing, due in part to ongoing efforts to develop new therapies for these disorders and new screening assays to identify increased risk for the individual MPSs on the basis of deficiency in the cognate enzyme. Existing tests for MPSs utilize either fluorescence or mass spectrometry detection methods to measure biomarkers of disease (e.g., enzyme function or glycosaminoglycans) using either urine or dried blood spot (DBS) samples. There are currently two approaches to fluorescence-based enzyme function assays from DBS: (1) manual reaction mixing, incubation, and termination followed by detection on a microtiter plate reader; and (2) miniaturized automation of these same assay steps using digital microfluidics technology. This article describes the origins of laboratory assays for enzyme activity measurement, the maturation and clinical application of fluorescent enzyme assays for MPS newborn screening, and considerations for future expansion of the technology. Full article

The present study aimed to compare the diagnostic accuracy between conventional smear (CS) and liquid-based preparation (LBP) in endoscopic ultrasonography-fine needle aspiration cytology (EUS-FNAC) of pancreatic lesions. Using 31 eligible studies, the diagnostic accuracy of cytologic examination in CS and LBP was evaluated through a conventional meta-analysis and diagnostic test accuracy review. Overall concordance rates were 82.8% (95% confidence interval [CI], 79.8–85.5%) and 94.0% (95% CI, 84.4–97.8%) in CS and LBP, respectively. CS with rapid on-site evaluation (ROSE) showed a higher concordance rate than CS without ROSE. In CS, the pooled sensitivity and specificity were 89.8% (95% CI, 85.2–93.1%) and 95.0% (95% CI, 90.0–97.6%), respectively. The diagnostic odds ratio (OR) and area under curve (AUC) of the summary receiver operating characteristic (SROC) curve were 90.32 (95% CI, 43.85–147.11) and 0.945, respectively. In LBP, the pooled sensitivity and specificity were 80.9% (95% CI, 69.7–88.7%) and 99.9% (95% CI, 1.5–100.0%), respectively. The diagnostic OR and AUC of the SROC curve were 57.21 (95% CI, 23.61–138.64) and 0.939, respectively. Higher concordance rates were found in CS with ROSE and LBP in EUS-FNAC of pancreatic lesions. Regardless of the cytologic preparation method, EUS-FNAC is a useful and accurate diagnostic tool for pancreatic lesions. Full article

Currently, mental stress is a common social problem affecting people. Stress reduces human functionality during routine work and may lead to severe health defects. Detecting stress is important in education and industry to determine the efficiency of teaching, to improve education, and to reduce risks from human errors that might occur due to workers’ stressful situations. Therefore, the early detection of mental stress using machine learning (ML) techniques is essential to prevent illness and health problems, improve quality of education, and improve industrial safety. The human brain is the main target of mental stress. For this reason, an ML system is proposed which investigates electroencephalogram (EEG) signal for thirty-six participants. Extracting useful features is essential for an efficient mental stress detection (MSD) system. Thus, this framework introduces a hybrid feature-set that feeds five ML classifiers to detect stress and non-stress states, and classify stress levels. To produce a reliable, practical, and efficient MSD system with a reduced number of electrodes, the proposed MSD scheme investigates the electrodes placements on different sites on the scalp and selects that site which has the higher impact on the accuracy of the system. Principal Component analysis is employed also, to reduce the features extracted from such electrodes to lower model complexity, where the optimal number of principal components is examined using sequential forward procedure. Furthermore, it examines the minimum number of electrodes placed on the site which has greater impact on stress detection and evaluation. To test the effectiveness of the proposed system, the results are compared with other feature extraction methods shown in literature. They are also compared with state-of-the-art techniques recorded for stress detection. The highest accuracies achieved in this study are 99.9%(sd = 0.015) and 99.26% (sd = 0.08) for identifying stress and non-stress states, and distinguishing between stress levels, respectively, using only two frontal brain electrodes for detecting stress and non-stress, and three frontal electrodes for evaluating stress levels respectively. The results show that the proposed system is reliable as the sensitivity is 99.9(0.064), 98.35(0.27), specificity is 99.94(0.02), 99.6(0.05), precision is 99.94(0.06), 98.9(0.23), and the diagnostics odd ratio (DOR) is ≥ 100 for detecting stress and non-stress, and evaluating stress levels respectively. This shows that the proposed framework has compelling performance and can be employed for stress detection and evaluation in medical, educational and industrial fields. Finally, the results verified the efficiency and reliability of the proposed system in predicting stress and non-stress on new patients, as the accuracy achieved 98.48% (sd = 1.12), sensitivity = 97.78% (sd = 1.84), specificity = 97.75% (sd = 2.05), precision = 99.26% (sd = 0.67), and DOR ≥ 100 using only two frontal electrodes. Full article

The typification of breast lumps with fine-needle biopsies is often affected by inconclusive results that extend diagnostic time. Many breast centers have progressively substituted cytology with micro-histology. The aim of this study is to assess the performance of a 13G-needle biopsy using cable-free vacuum-assisted breast biopsy (VABB) technology. Two of our operators carried out 200 micro-histological biopsies using the Elite 13G-needle VABB and 1314 14–16G-needle core biopsies (CBs) on BI-RADS 3, 4, and 5 lesions. Thirty-one of the procedures were repeated following CB, eighteen following cytological biopsy, and three after undergoing both procedures. The VABB Elite procedure showed high diagnostic performance with an accuracy of 94.00%, a sensitivity of 92.30%, and a specificity of 100%, while the diagnostic underestimation was 11.00%, all significantly comparable to of the CB procedure. The VABB Elite 13G system has been shown to be a simple, rapid, reliable, and well-tolerated biopsy procedure, without any significant complications and with a diagnostic performance comparable to traditional CB procedures. The histological class change in an extremely high number of samples would suggest the use of this procedure as a second-line biopsy for suspect cases or those with indeterminate cyto-histological results. Full article

The coronavirus disease 2019 (COVID-19) outbreak, caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global ongoing pandemic. Timely, accurate and non-invasive SARS-CoV-2 detection in both symptomatic and asymptomatic patients, as well as determination of their immune status, will facilitate effective large-scale pandemic control measures to prevent the spread of COVID-19. Saliva is a biofluid whose anatomical source and location is of particularly strategic relevance to COVID-19 transmission and monitoring. This review focuses on the role of saliva as both a foe (a common mode of viral transmission via salivary droplets and potentially aerosols) and a friend (as a non-invasive diagnostic tool for viral detection and immune status surveillance) in combating COVID-19. Full article

A fully automatic method for specific binding ratio (SBR) calculation in [¹²³I]ioflupane single-photon emission computed tomography (SPECT) studies was proposed by creating volumes of interest of the striatum (VOI_st) and reference region (VOI_ref) without manual handling to avoid operator-induced variability. The study involved 105 patients (72 ± 10 years) suspected of parkinsonian syndrome (PS) who underwent [¹²³I]ioflupane SPECT. The 200 images from our previous study were used for evaluation and validation of the new program. All patients were classified into PS and non-PS groups according to the results of clinical follow-up. A trapezoidal volume of interest (VOI_t) containing all striatal intensive counts was created automatically, followed by VOI_st setting using the previous method. SBR values were calculated from the mean values of VOI_st and VOI_ref determined by the whole brain outside of VOI_t. The low count voxels in the VOI_ref were excluded using an appropriate threshold. The SBR values from the new method were compared with the previous semi-automatic method and the Tossici–Bolt (TB) method. The SBRs from the semi- and fully automatic methods showed a good linear correlation (r > 0.98). The areas under the curves (AUCs) of receiver operating characteristic analysis showed no significant difference between the two methods for both our previous (AUC > 0.99) and new (AUC > 0.95) data. The diagnostic accuracy of the two methods showed similar results (>92%), and both were better than the TB method. The proposed method successfully created the automatic VOIs and calculated SBR rapidly (9 ± 1 s/patient), avoiding operator-induced variability and providing objective SBR results. Full article

A new generation of magnetic lateral flow immunoassays is emerging as powerful tool for diagnostics. They rely on the use of magnetic nanoparticles (MNP) as detecting label, replacing conventional gold or latex beads. MNPs can be sensed and quantified by means of external devices, allowing the development of immunochromatographic tests with a quantitative capability. Moreover, they have an added advantage because they can be used for immunomagnetic separation (IMS), with improvements in selectivity and sensitivity. In this paper, we have reviewed the current knowledge on magnetic-lateral flow immunoassay (LFIA), coupled with both research and commercially available instruments. The work in the literature has been classified in two categories: optical and magnetic sensing. We have analysed the type of magnetic nanoparticles used in each case, their size, coating, crystal structure and the functional groups for their conjugation with biomolecules. We have also taken into account the analytical characteristics and the type of transduction. Magnetic LFIA have been used for the determination of biomarkers, pathogens, toxins, allergens and drugs. Nanocomposites have been developed as alternative to MNP with the purpose of sensitivity enhancement. Moreover, IMS in combination with other detection principles could also improve sensitivity and limit of detection. The critical analysis in this review could have an impact for the future development of magnetic LFIA in fields requiring both rapid separation and quantification. Full article

Among 2085 asymptomatic subjects (age: 51.0 ± 10.7 years, 41.3% female) with data available on common carotid artery diameter (CCAD) and circulating total white blood cell (WBC) counts, higher circulating leukocytes positively correlated with higher high sensitivity C-reactive protein (hs-CRP). Higher WBC/segmented cells and monocyte counts were independently associated with greater relative wall thicknesses and larger CCADs, which in general were more pronounced in men and obese subjects (body mass index ≥ 25 kg/m²) (all P _interaction: < 0.05). Using multivariate adjusting models, only the monocyte count independently predicted the left ventricular mass index (LVMi) (ß-Coef: 0.06, p = 0.01). Higher circulating WBC, segmented, and monocyte counts and a greater CCAD were all independently associated with a higher risk of heart failure (HF)/all-cause death during a median of 12.1 years of follow-up in fully adjusted models, with individuals manifesting both higher CCADs and monocyte counts incurring the highest risk of HF/death (adjusted hazard ratio: 2.81, 95% CI: 1.57. −5.03, p < 0.001; P _interaction, 0.035; lower CCAD/lower monocyte as reference). We conclude that a higher monocyte count is associated with cardiac remodeling and carotid artery dilation. Both an elevated monocyte count and a larger CCAD may indicate a specific phenotype that confers the highest risk of HF, which likely signifies the role of circulating monocytes in the pathophysiology of heart failure with preserved ejection fraction (HFpEF). Full article

Skeletal dysplasia (SD) is a complex group of bone and cartilage disorders often detectable by fetal ultrasound, but the definitive diagnosis remains challenging because the phenotypes are highly variable and often overlap among different disorders. The molecular mechanisms underlying this condition are also diverse. Hundreds of genes are involved in the pathogenesis of SD, but most of them are yet to be elucidated, rendering genotyping almost infeasible except those most common such as fibroblast growth factor receptor 3 (FGFR3), collagen type I alpha 1 chain (COL1A1), collagen type I alpha 2 chain (COL1A2), diastrophic dysplasia sulfate transporter (DTDST), and SRY-box 9 (SOX9). Here, we report the use of trio-based whole exome sequencing (trio-WES) with comprehensive gene set analysis in two Taiwanese non-consanguineous families with fetal SD at autopsy. A biparental-origin homozygous c.509G>A(p.G170D) mutation in peptidylprolyl isomerase B (PPIB) gene was identified. The results support a diagnosis of a rare form of autosomal recessive SD, osteogenesis imperfecta type IX (OI IX), and confirm that the use of a trio-WES study is helpful to uncover a genetic explanation for observed fetal anomalies (e.g., SD), especially in cases suggesting autosomal recessive inheritance. Moreover, the finding of an identical PPIB mutation in two non-consanguineous families highlights the possibility of the founder effect, which deserves future investigations in the Taiwanese population. Full article

In this paper, we present an architecture of a personalized glucose monitoring system (PGMS). PGMS consists of both invasive and non-invasive sensors on a single device. Initially, blood glucose is measured invasively and non-invasively, to train the machine learning models. Then, paired data and corresponding errors are divided scientifically into six different clusters based on blood glucose ranges as per the patient’s diabetic conditions. Each cluster is trained to build the unique error prediction model using an adaptive boosting (AdaBoost) algorithm. Later, these error prediction models undergo personalized calibration based on the patient’s characteristics. Once, the errors in predicted non-invasive values are within the acceptable error range, the device gets personalized for a patient to measure the blood glucose non-invasively. We verify PGMS on two different datasets. Performance analysis shows that the mean absolute relative difference (MARD) is reduced exceptionally to 7.3% and 7.1% for predicted values as compared to 25.4% and 18.4% for measured non-invasive glucose values. The Clarke error grid analysis (CEGA) plot for non-invasive predicted values shows 97% data in Zone A and 3% data in Zone B for dataset 1. Moreover, for dataset 2 results echoed with 98% and 2% in Zones A and B, respectively. Full article

This study was conducted to investigate if anthropometric features can predict the depth of the soleus muscle, as assessed with ultrasound imaging, in a sample of healthy individuals to assist clinicians in the application of dry needling. A diagnostic study to calculate the accuracy of a prediction model for soleus muscle depth, as assessed with ultrasonography, in the middle-third and distal-third of the calf, based on anthropometric features such as age, height, weight, body mass index (BMI), calf length, mid-third and distal-third calf girth, was conducted on 48 asymptomatic healthy subjects (75% male) involving a total of 96 calves. Multiple linear regression analyses were used to determine which variables contributed significantly to the variance in the soleus muscle depth at middle-third and distal-third of the calf by gender. Women were found to have a deeper soleus muscle than men (p < 0.001). Weight, height, BMI, and mid-third calf perimeter explained 69.9% of variance in men, whereas mid-third calf perimeter, calf length, height, and distal-third calf girth explained 73% of the variance in women of the distal-third soleus depth (p < 0.001). Additionally, mid-third calf girth and calf length explained 28.8% of variance in men, whereas mid-third calf perimeter, calf length, and weight explained 67.8% of variance in women of the mid-third soleus depth (p < 0.001). This study identified anthropometric features that predict soleus muscle depth, as assessed with ultrasound, in asymptomatic individuals, but these features are different in men and women. Our findings could assist clinicians in choosing the proper length of the needle to avoiding passing through the soleus during dry needling. Full article

Severe acute respiratory syndrome-Coronavirus-2 (SARS-CoV-2) is a novel viral infection characterized by several symptoms range from mild to severe clinical conditions that could lead to death. We report two different radiological findings on computed tomography (CT) in two patients affected by SARS-CoV-2: a lung acute embolism (APE) in the first case and a radiological picture of acute respiratory distress syndrome (ARDS) in the second case. This is an important issue to be identified in order to provide more specific therapy earlier, including both antiviral and anti-inflammatory drugs associated with anti anticoagulant therapy. Full article

Background: Cystic fibrosis (CF) is a hereditary disorder in which persistent unresolved inflammation and recurrent airway infections play major roles in the initiation and progression of the disease. Little is known about triggering factors modulating the transition to chronic microbial infection and inflammation particularly in young children. Cystic fibrosis respiratory disease starts early in life, with the detection of inflammatory markers and infection evident even before respiratory symptoms arise. Thus, identifying factors that dysregulate immune responsiveness at the earliest stages of the disease will provide novel targets for early therapeutic intervention. Methods: We evaluated the clinical significance of bile acid detection in the bronchoalveolar lavage fluid of clinically stable preschool-aged children diagnosed with CF. Results: We applied an unbiased classification strategy to categorize these specimens based on bile acid profiles. We provide clear associations linking the presence of bile acids in the lungs with alterations in the expression of inflammatory markers. Using multiple regression analysis, we also demonstrate that clustering based on bile acid profiles is a meaningful predictor of the progression of structural lung disease. Conclusions: Altogether, our work has identified a clinically relevant host-derived factor that may participate in shaping early events in the aetiology of CF respiratory disease. Full article

Hysteroscopy has known an increasing use in Romania over the last decade, succeeding to mark an impact on lowering the costs of medical services. The strategy of this study was to present the experience of two Romanian medical units with different experiences using inpatient regimen hysteroscopy, and to further compare it with current worldwide tendencies. Strong points in our practice were sought, as well as components that require improvement. Overall, abnormal uterine bleeding stands for most hospital case presentations in our study group; hysteroscopy had the highest accuracy and positive predictive value in identifying uterine myomas: 91.03% and 100%, respectively; for endometrial polyps, statistical analysis showed the highest sensitivity: 100%, with 83.89% specificity and a 77.64% positive predictive value. The applicability of hysteroscopy was further described for infertility cases and isthmocele repair. Full article