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The Fontan operation has become the primary palliative treatment for patients with a functionally univentricular heart. The population of patients with Fontan circulation is constantly growing and aging. As the number of Fontan patients surviving into adulthood increases, there is a clear need for research on how best to follow these patients and manage their complications. Monitoring blood biomarkers is a promising method for the non-invasive assessment of the Fontan circulation. In this article, we provide a comprehensive review of the available evidence on this topic. The following biomarkers were included: natriuretic peptides, red blood cell distribution width (RDW), cystatin C, high-sensitivity C-reactive protein, vitamin D, parathyroid hormone, von Willebrand factor, carbohydrate antigen 125, lipoproteins, hepatocyte growth factor, troponins, ST2 protein, galectin-3, adrenomedullin, endothelin-1, components of the renin-angiotensin-aldosterone system, norepinephrine, interleukin 6, tumor necrosis factor α, and uric acid. We did not find strong enough data to propose evidence-based recommendations. Nevertheless, significantly elevated levels of brain natriuretic peptide (BNP)/N-terminal prohormone of BNP (NT-proBNP) are most likely associated with the failure of the Fontan circulation. The use of the RDW is also promising. Several biomarkers appear to be useful in certain clinical presentations. Certainly, robust longitudinal, preferably multicenter, prospective studies are needed to determine the sensitivity, specificity, evidence-based cut-off values and overall predictive value of different biomarkers in monitoring Fontan physiology.

Purpose: Human presence in space is increasingly frequent, but we must not forget that it is a hostile environment. We aimed to study the characteristics of experimental scenarios, to obtain data on human response to isolation, disruption of circadian rhythm and high levels of psychophysical stress., Methods: In these experiments, we evaluated stress response in five young healthy subjects inside an earth-based moon-settlement-like habitat during a 1-week long analog astronaut mission. Wearable devices were used to monitor daily step count of the subjects, physical activity, heart rate during physical exercise and at rest, and sleep parameters. From saliva and urine samples collected every day at awakening, we studied oxy-inflammation biomarkers and hormones (stress and appetite) were studied too., Results: At the end of the week, all subjects revealed an increase in oxidative stress and cortisol levels but no inflammation biomarkers variations, in conjunction with increasing time/daily exercise. Furthermore, a significant decrease in hours of sleep/day, sleep quality, and REM phase of sleep was recorded and correlated with the increase of reactive oxygen species., Conclusion: Oxidative stress increased in a short period of time and may be attributed to the influence of psychological stress during confinement, as well as increased exercise and decreased amount of sleep. On a long-term basis, this could impact performance., Competing Interests: Declarations. Conflict of interest: The authors report no competing interests., (© 2024. The Author(s).)

Cortisol, an essential glucocorticoid hormone, is crucial in regulating the stress response and maintaining physiological and behavioral homeostasis in mammals, including dogs. This review explores cortisol's physiological and behavioral role in canines, focusing on its effects on stress, immune function, and metabolism. Various methods of measuring cortisol levels in dogs, invasive (blood, saliva, urine) and non-invasive (hair, fecal assays), are discussed regarding their accuracy and practical applications. The review also highlights the influence of different environmental factors, such as shelter conditions, human interaction, and music, on cortisol levels in dogs. Furthermore, the clinical and behavioral implications of abnormal cortisol levels are examined, with particular attention to conditions like Cushing's disease and stress-related behavioral issues. The findings emphasize the importance of cortisol monitoring in veterinary practice and animal welfare, proposing future research directions to improve canine health and stress management.

The continuous search for simple, noninvasive methods for assessing liver fibrosis remains very important to help risk-stratify and follow-up patients with chronic hepatitis C virus (HCV). This study aimed to evaluate the diagnostic performance and accuracy of six serological noninvasive scores for the assessment of liver fibrosis in comparison to liver histopathology. This retrospective cohort study included data from 19501 patients with chronic HCV infection who had liver biopsies as an HCV treatment prerequisite within the Egyptian national HCV treatment program. Six noninvasive scores (FIB-4, APRI, King's score, Fibro-Q, fibrosis index, Fibro-α score) were evaluated and compared to liver histopathology data in assessing different stages of liver fibrosis. The diagnostic performance for each score was assessed using the area under the receiver-operating characteristic curve (AUROC). All six noninvasive scores were statistically significant for predicting different stages of liver fibrosis. Four scores (FIB-4, King's score, APRI, and Fibro Q) had a better diagnostic performance for predicting different fibrosis stages. FIB-4, followed by the King's score, performs better in identifying patients with advanced fibrosis at cutoffs of 2.01 and 16.7, respectively, with AUROC of 0.71 for both, and in predicting cirrhosis at cutoffs of 2.21 and 17.4, respectively with AUROC 0.82 for both. Using noninvasive scores for fibrosis assessment is very important, especially in limited resource settings, to rapidly stratify patients who need more specialized care., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Multispectral devices have a huge potential to be utilized in biological, ecological, and agricultural studies, providing valuable information on plant structure and chemical composition. The aim of the study was to assess the reliability and sensitivity of the affordable leaf spectrometer PolyPen (PP) in comparison with the highly sensitive analytical device FieldSpec-4. Measurements at the leaf level were realized on a collection of 24 diverse field-grown wheat ( Triticum sp. L.) genotypes in several growth phases during the regular growing season, focusing on whole spectral curves and a set of 41 spectral reflectance indices. As expected, the sensitive analytical device showed a higher capacity to capture genotypic variability and the ability to distinguish seasonal changes compared to a low-cost multispectral device. Nevertheless, the analysis of the data provided by low-cost sensors provided a group of parameters with good sensitivity, including reasonable correlations between the records of the two devices ( r > 0.80). Based on the large obtained datasets, we can conclude that the application of a low-cost PP leaf spectrometer in plant and crop studies can be efficient, but the selection of parameters is crucial. Thus, the present study provides valuable information for users of affordable leaf spectrometers in fundamental and applied plant science., Competing Interests: Conflict of interest: Authors state no conflict of interest., (© 2024 the author(s), published by De Gruyter.)

The common octopus ( Octopus vulgaris) is an excellent candidate for aquaculture diversification, due to its biological traits and high market demand. To ensure a high-quality product while maintaining welfare in captive environments, it is crucial to develop non-invasive methods for testing health biomarkers. Proteins found in skin mucus offer a non-invasive approach to monitoring octopus welfare. This study compares the protein profiles in the skin mucus of wild, aquarium-maintained, and senescent specimens to identify welfare biomarkers. A tandem mass tag (TMT) coupled with an Orbitrap Eclipse Tribrid mass spectrometer was used to create a reference dataset from octopus skin mucus, identifying 1496 non-redundant protein groups. Although similar profiles were observed, differences in relative abundances led to the identification of potential biomarkers, including caspase-3-like, protocadherin 4, deleted in malignant brain tumors, thioredoxin, papilin, annexin, cofilin and mucin-4 proteins. Some of these proteins also revealed potential as bioactive peptides. This investigation provides the most extensive analysis of the skin mucus proteome in the common octopus and is the first to explore how aquarium maintenance and senescence alter the mucus proteome. This research highlights the potential of skin mucus protein/peptides as non-invasive monitoring biomarkers in cultured animals.

Recent advancements in asthma management include non-invasive methodologies such as sputum analysis, exhaled breath condensate (EBC), and fractional exhaled nitric oxide (FeNO). These techniques offer a means to assess airway inflammation, a critical feature of asthma, without invasive procedures. Sputum analysis provides detailed insights into airway inflammation patterns and cellular composition, guiding personalized treatment strategies. EBC collection, reflecting bronchoalveolar lining fluid composition, provides a non-invasive window into airway physiology. FeNO emerges as a pivotal biomarker, offering insights into eosinophilic airway inflammation and aiding in asthma diagnosis, treatment monitoring, and the prediction of exacerbation risks. Despite inherent limitations, each method offers valuable tools for a more comprehensive assessment of asthma. Combining these techniques with traditional methods like spirometry may lead to more personalized treatment plans and improved patient outcomes. Future research is crucial to refine protocols, validate biomarkers, and establish comprehensive guidelines in order to enhance asthma management with tailored therapeutic strategies and improved patient outcomes.

Background: Chronic hepatitis B (CHB) presents a global health challenge due to its potential to cause severe liver conditions such as hepatocellular carcinoma (HCC) and cirrhosis. Prior research has established a correlation between CHB infection with low-level viremia (LLV) and liver disease progression, such as increased HCC incidence. This study aims to investigate whether LLV during treatment with nucleos(t)ide analogs (NAs) contributes to the accelerated progression of liver fibrosis (LF)., Methods: This retrospective cohort study at Jinhua Central Hospital focused on CHB patients undergone NA monotherapy for over 96 weeks. Patients were categorized into maintained virological response (MVR) and LLV groups based on hepatitis B virus (HBV) DNA levels. The study assessed LF using various markers and methods, including chitinase 3-like 1 protein (CHI3L1), aspartate aminotransferase-to-platelet ratio index (APRI), fibrosis-4 (FIB-4) score, and transient elastography., Results: Analysis was conducted on 92 CHB patients, categorized into LLV (n=42) and MVR (n=50) groups, following the exclusion of 101 patients for various reasons. Significant findings included lower baseline HBV DNA in MVR (<20 IU/mL) compared to LLV (67.8 IU/mL, P<0.001) and different AST/ALT ratios (LLV: 1.1, MVR: 1.36, P=0.011). LF was assessed using CHI3L1, FIB-4, and APRI, with LLV showing a higher baseline CHI3L1 (LLV:83.3 ng/mL vs MVR: 54.5 ng/mL, P=0.016) and scores compared to MVR, indicative of fibrosis. CHI3L1 levels in LLV were higher at baseline and weeks 48, 72, and 96 than MVR, with significance at baseline (P=0.038) and week 48 (P=0.034). Liver stiffness measurement (LSM) showed a time-dependent decline in both groups but no significant intergroup differences., Conclusion: Non-invasive monitoring of CHB patients who have received treatment indicates that LLV contributes to the progression of LF, necessitating proactive adjustment of antiviral treatment strategies., Competing Interests: The authors declare that they have no competing interests in this work., (© 2024 Xu et al.)

Introduction: Intensified hand hygiene measures were recommended for preventing the spread of SARS-CoV-2. However, these measures can lead to skin damage and the development of hand eczema, particularly among health professionals., Objectives: This pilot study aimed to evaluate the effects of repeated antiseptic use on healthy skin under controlled conditions and to assess the emollient use., Methods: Twelve healthy volunteers (nine females, age = 22.3 ± 2.8 years (mean ± SD), Fitzpatrick phototypes II and III) with no skin diseases were recruited. Antiseptic was applied daily for 3 weeks on the volar sides of forearms. Emollient cream was also applied daily. Skin assessments were performed using non-invasive methods (transepidermal water loss-TEWL, skin hydration, erythema and melanin content)., Results: Prolonged antiseptic use increased TEWL, decreased hydration and elevated erythema and melanin levels. Emollient cream significantly reduced TEWL and improved hydration on antiseptic-treated sites, and also enhanced hydration on intact skin., Conclusions: Prolonged use of antiseptics can have adverse effects on the skin, including barrier disruption and inflammation. Emollient showed promise in improving skin hydration and reducing the damage caused by antiseptics. Further research with a larger sample is needed to confirm these findings and assess emollient efficacy during frequent antiseptic use., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Skin mucus analysis has recently been used as a non-invasive method to evaluate for fish welfare. The present research study was conducted to examine the skin mucosal immunity and skin microbiota profiles of sturgeons infected with Citrobacter freundii. Our histology results showed that the thickness of the epidermal layer of skin remained thinner, and the number of mucous cells was significantly decreased in sturgeons after infection (p < 0.05). Total protein, alanine aminotransferase, aspartate aminotransferase, superoxide dismutase, and creatine kinase levels in the mucus showed biphasic pattern (decrease and then increase). Lactate dehydrogenase, lysozyme, and acid phosphatase activities in the mucus showed an increasing trend after infection. Furthermore, 16S rRNA sequencing also revealed that C. freundii infection also affected the diversity and community structure of the skin mucus microbiota. An increase in microbial diversity (p > 0.05) and a decrease in microbial abundance (p < 0.05) after infection were noted. The predominant bacterial phyla in the skin mucus were Proteobacteria, Fusobacteria, Bacteroidetes, Firmicutes, and Actinobacteria. Specifically, the relative abundance of Fusobacteria increased after infection. The predominant bacterial genera in the skin mucus were Cetobacterium, Pelomonas, Bradyrhizobium, Flavobacterium, and Pseudomonas. The relative abundance of Cetobacterium, Pseudomonas, and Flavobacterium increased after infection. Our current research findings will provide new insights into the theoretical basis for future research studies exploring the mechanism of sturgeon infection with C. freundii., Competing Interests: Declaration of competing interest The authors declare no known financial or personal interests that could have influenced the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Solid organ transplantation is a life-saving intervention for individuals with end-stage organ failure. Despite the effectiveness of immunosuppressive therapy, the risk of graft rejection persists in all viable transplants between individuals. The risk of rejection may vary depending on the degree of compatibility between the donor and recipient for both human leucocyte antigen (HLA) and non-HLA gene-encoded products. Monitoring the status of the allograft is a critical aspect of post-transplant management, with invasive biopsies being the standard of care for detecting rejection. Non-invasive biomarkers are increasingly being recognized as valuable tools for aiding in the detection of graft rejection, monitoring graft status and evaluating the efficacy of immunosuppressive therapy. Here, we focus on the importance of molecular biomarkers in solid organ transplantation and their potential role in clinical practice. Conventional molecular biomarkers used in transplantation include HLA typing, detection of anti-HLA antibodies, killer cell immunoglobulin-like receptor genotypes, and anti-MHC class 1-related chain A antibodies, which are important for assessing the compatibility of the donor and recipient. Emerging molecular biomarkers include the detection of donor-derived cell-free DNA, microRNAs (regulation of gene expression), exosomes (small vesicles secreted by cells), and kidney solid organ response test, in the recipient's blood for early signs of rejection. This review highlights the strengths and limitations of these molecular biomarkers and their potential role in improving transplant outcomes., (© 2024 John Wiley & Sons Ltd.)

The application of faecal DNA in genetic studies of wild populations minimises disturbances to their normal behaviours and body integrity. Here, I present an analysis of a metagenomic dataset generated from the faecal DNA of several specimens of the estuarine pipefish, Syngnathus watermeyeri, to simultaneously assemble the mitogenomes of the predator and its main prey species, the copepod Pseudodiaptomus hessei . The mitogenomes of the pipefish and the copepod were successfully reconstructed using a combination of short seed extension and denovo metagenomic assembly. Nucleotide blast searches of the circular contigs, mitogenome annotations, and Bayesian phylogenetic analyses confirm the completeness and correct taxonomic placements of the two mitogenomes. In addition, heteroplasmy detection and Pool-Seq variant calling quantified the level of genetic diversity in the sequences that formed these assemblies. These can be used as a first step to non-invasively survey genetic diversity in these populations., (© 2023 The Author(s).)

Non-invasive methods are important to the field of conservation physiology to reduce negative effects on organisms being studied. Glucocorticoid (GC) hormones are often used to assess health of individuals, but collection methods can be invasive. Many amphibians are imperiled worldwide, and saliva is a non- or semi-invasive matrix to measure GCs that has been partially validated for only four amphibian species. Validation ensures that assays are reliable and can detect changes in saliva corticosterone (sCORT) after exposure to stressors, but it is also necessary to ensure sCORT concentrations are correlated with plasma concentrations. To help validate the use of saliva in assessing CORT responses in amphibians, we captured uniquely marked Columbia spotted frogs ( Rana luteiventris ) on sequential days and collected baseline and stress-induced (after handling) samples. For a subset of individuals, we collected and quantified CORT in both saliva and blood samples, which have not been compared for amphibians. We tested several aspects of CORT responses and, by collecting across separate days, measured repeatability of CORT responses across days. We also evaluated whether methods common to amphibian conservation, such as handling alone or handling, clipping a toe and tagging elevated sCORT. Similar to previous studies, we show that sCORT is reliable concerning parallelism, recovery, precision and sensitivity. sCORT was weakly correlated with plasma CORT (R 2  = 0.21), and we detected elevations in sCORT after handling, demonstrating biological validation. Toe clipping and tagging did not increase sCORT over handling alone, but repeated handling elevated sCORT for ~72 hours. However, sCORT responses were highly variable and repeatability was low within individuals and among capture sessions, contrary to previous studies with urinary and waterborne CORT. sCORT is a semi-invasive and rapid technique that could be useful to assess effects of anthropogenic change and conservation efforts, but will require careful study design and future validation., (© The Author(s) 2023. Published by Oxford University Press and the Society for Experimental Biology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Long COVID-19 patients show systemic inflammation and persistent symptoms such as fatigue and malaise, profoundly affecting their quality of life. Since improving oxygenation can oppose inflammation at multiple tissue levels, we hypothesized that hyperbaric oxygen therapy (HBOT) could arrest inflammation progression and thus relieve symptoms of COVID-19. We evaluated oxy-inflammation biomarkers in long COVID-19 subjects treated with HBOT and monitored with non-invasive methods. Five subjects (two athletes and three patients with other comorbidities) were assigned to receive HBOT: 100% inspired O 2 at 2.4 ATA in a multiplace hyperbaric chamber for 90 min (three athletes: 15 HBOT × 5 days/wk for 3 weeks; two patients affected by Idiopathic Sudden Sensorineural Hearing Loss: 30 HBOT × 5 days/wk for 6 weeks; and one patient with osteomyelitis: 30 HBOT × 5 days/wk for week for 6 weeks and, after a 30-day break, followed by a second cycle of 20 HBOT). Using saliva and/or urine samples, reactive oxygen species (ROS), antioxidant capacity, cytokines, lipids peroxidation, DNA damage, and renal status were assessed at T1&#95;pre (basal level) and at T2&#95;pre (basal level after treatment), and the results showed attenuated ROS production, lipid peroxidation, DNA damage, NO metabolites, and inflammation biomarker levels, especially in the athletes post-treatment. Thus, HBOT may represent an alternative non-invasive method for treating long COVID-19-induced long-lasting manifestations of oxy-inflammation.

Monitoring the conservation status of endangered freshwater fish using less invasive methods poses challenges for ecologists and conservationists. Visual surveys have been proposed as an alternative to electrofishing, which is a standard methodology that can cause injuries, physiological stress and post-release mortality in organisms. To test the efficacy of visual methods, a study was conducted in an intermittent stream of Sardinia (Italy). Two visual methods were employed: a visual survey from streambanks (VSS) and an underwater visual survey (UVS) using cameras. The aims of this study were (1) to compare the effectiveness of these methods in detecting patch occupancy patterns and (2) to investigate the effect of environmental variables on the detection probability of Mediterranean native trout. Environmental variables characterizing pool habitats were recorded, and generalized linear models (GLMs) were employed to assess the correlation between these variables and trout presence/absence. GLM analysis revealed that UVS had higher detection probability with larger pool volume, whereas submerged macrophytes negatively affected detection probability. Detection from streambanks (VVS) was negatively affected by a high turbulence rate. In conclusion, our study suggests the utility of visual methods to describe patterns of patch occupancy of Mediterranean trout. However, methods can be differently affected by environmental variables. Therefore, monitoring programs using these methods should consider these factors to ensure a reliable description of within-stream trout distribution in intermittent streams.

Objective: The aim: To determine the diagnostic value of non-invasive methods of GERD diagnosis based on questionnaire data and a diagnostic test with a proton pump inhibitor (PPI) with Rabeprazole in patients with coronary heart disease (CHD)., Patients and Methods: Materials and methods: 90 patients were under observation, namely, 68 patients with coronary heart disease with concomitant essential arterial hyperten¬sion (EAH), 6 patients with coronary artery disease + arrhythmias, and 18 - others diseases. All patients were surveyed according to the GerdQ questionnaire, followed by PPI testing with Rabeprazole, and body mass index (BMI) was calculated for all the patients. A BMI of 18.5-24.9 kg/m2 was considered as normal body weight, and a BMI > 24.9 kg/m2 was considered overweight., Results: Results: Based on the results of the questionnaire, it was established that the most common complaints typical for GERD were noted by patients with CHD in combination with EAH. There were 48 of such patients and, depending on BMI, they were divided into 2 groups: 1st group (n=14) - patients with CHD+EAH+GERD with normal body weight and 2nd group (n=34) - overweight patients with CHD+EAH+GERD. In patients of group 1, typical symptoms of GERD prevailed (in 71.4% of patients), and in patients group 2, the distribution of typical and extra-esophageal symptoms did not differ significantly (52.9% vs. 47.1%). Among the extraesophageal manifestations, pain behind the sternum (in the projection of the esophagus) was significantly more often recorded in patients of group 1, and rhythm disturbances in patients of group 2 (43.8% and 75.0% of patients, respectively, p<0.05). The results of the GerdQ questionnaire showed a direct relationship between GERD, body weight and symptom score. Patients with GERD+normal body weight had a mean score of 6 for classic gastroesophageal reflux symptoms, while patients with GERD+increased body weight had a mean score of 7. The sensitivity of the questionnaire was 78.7%, and the specificity - 92.9%. According to the PPI test, in the 1st and 2nd groups, already in the first three days, 28.5% and 23.5% of patients noted the disappearance of heartburn and after 10 days - 85.7% and 64.7%, respectively. Over the entire period, that is, after 14 days of observation, 85.7% of patients in the 1st group and 73.5% in the 2nd group noted improvement., Conclusion: Conclusions: It has been established that questionnaires based on the GerdQ questionnaire followed by a PPI test with Rabeprazole in patients with coronary heart disease combined with essential arterial hypertension have a high diagnostic value and can be used for early diagnosis and effective treatment of GERD.

According to World Health Organization reports, large numbers of people around the globe have been infected or died for Covid-19 due to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Researchers are still trying to find a rapid and accurate diagnostic method for revealing infected people by low viral load with the overriding goal of effective diagnostic management. Monitoring the body metabolic changes is known as an effective and inexpensive approach for the evaluation of the infected people. Here, an optical sniffer is introduced to detect exhaled breath metabolites of patients with Covid-19 (60 samples), healthy humans (55 samples), and cured people (15 samples), providing a unique color pattern for differentiation between the studied samples. The sniffer device is installed on a thin face mask, and directly exposed to the exhaled breath stream. The interactions occurring between the volatile compounds and sensing components such as porphyrazines, modified organic dyes, porphyrins, inorganic complexes, and gold nanoparticles allowing for the change of the color, thus being tracked as the sensor responses. The assay accuracy for the differentiation between patient, healthy and cured samples is calculated to be in the range of 80%-84%. The changes in the color of the sensor have a linear correlation with the disease severity and viral load evaluated by rRT-PCR method. Interestingly, comorbidities such as kidney, lung, and diabetes diseases as well as being a smoker may be diagnosed by the proposed method. As a powerful detection device, the breath sniffer can replace the conventional rapid test kits for medical applications., (© 2022 Elsevier B.V. All rights reserved.)

Purpose: We conducted the first meta-analysis to determine the diagnostic value of non-invasive methods for diagnosing bladder outlet obstruction (BOO) in men with lower urinary tract symptoms (LUTS)., Methods: We searched a range of databases for relevant publications up to June 2022, including PubMed, Embase, Web of Science, and the Cochrane Library. Retrieved studies were then reviewed for eligibility and data were extracted. The risk of bias (RoB) was assessed using the QUADAS-2 tool. We then performed a formal meta-analysis to evaluate the accuracy of various non-invasive methods for diagnosing BOO in men., Results: We identified 51 eligible studies including 7,897 patients for meta-analysis. The majority of the studies had a low overall RoB. Detrusor wall thickness (DWT) (pooled sensitivity (SSY): 71%; specificity (SPY): 88%; diagnostic odds ratio (DOR): 17.15; area under curve (AUC) 0.87) and the penile cuff test (PCT) (pooled SSY: 87%; SPY: 78%; DOR: 23.54; AUC: 0.88) showed high accuracy for diagnosing BOO. Furthermore, data suggested that DWT had the highest pooled SPY (0.89), DOR (32.58), and AUC (0.90), when using 2 mm as the cut-off., Conclusion: Of the non-invasive tests tested, DWT and PCT had the highest levels of diagnostic accuracy for diagnosing BOO in men with LUTS. DWT, with a 2 mm cut-off, had the highest level of accuracy. These two methods represent good options as non-invasive tools for evaluating BOO in males., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer MX declared a shared affiliation, with no collaboration, with the authors at the time of review., (© 2022 Cheng, Li, Wu, Ling, Rao, Yuan, Chen, Du and Xu.)

This report describe the first application of environmental DNA-metabarcoding approach for the assessment of fish species diversity in two marine protected areas of the North Sea: the Doggerbank and the Sylt Outer Reef. We collected 64 water samples and detected 24 fish species. We discuss qualitative differences between MPAs and compare the results with those obtained from bottom-trawl surveys in the same areas. We found three additional species to those documented in the same year with trawls, including the critically endangered European eel., (© 2022 Fisheries Society of the British Isles.)

Background: Data concerning hepatitis C virus (HCV) treatment using direct-acting agents (DAAs) post liver transplantation (LT) remains scarce in low- and average-income countries., Aim of the Study: To evaluate the safety and efficacy of post-LT HCV treatment using DAAs in Rio de Janeiro (Brazil), and to assess the course of hepatic biomarkers after sustained virological response (SVR)., Methods: Data from LT recipients with recurrent HCV treated using DAAs was retrospectively analyzed. HCV was defined by detectable HCV-RNA with elevated aminotransferases and/or histological signs of infection on liver biopsy post LT. SVR was defined as undetectable HCV-RNA 12 weeks after the end of treatment. Aspartate-to-Platelet Ratio Index (APRI) and Fibrosis-4 score (FIB-4) were calculated before treatment and after SVR., Results: 116 patients (63% male, median age 62 years, 75% genotype 1 and 62% with hepatocellular carcinoma [HCC] prior to LT) were included. Cirrhosis was identified in the allograft of 21 subjects (18%). The overall SVR was 96.6% without differences in SVR proportion according to clinical/demographic characteristics, genotype or presence of cirrhosis. SVR rates were similar in individuals with and without HCC pre-LT (95.8% [95% CI: 87.6-98.7] vs. 97.7% [95% CI: 85.0-99.7%], p = 0.588). No serious adverse events were observed and the use of ribavirin was associated with at least one adverse event (OR = 8.71 [95% CI: 3.17-23.99]). SVR was associated with regression of APRI (OR = 26.00 [95% CI 4.27-1065.94]) and FIB-4 (OR = 15.00 [95% CI: 2.30-631.47])., Conclusion: Post-LT HCV treatment with DAAs was safe and effective and associated with a significant decrease in hepatic biomarker levels after SVR., Competing Interests: Conflict of Interest The authors have no conflicts of interest to disclose related to this topic., (Copyright © 2022. Published by Elsevier Inc.)

In the last decades, wildlife diseases and the health status of animal populations have gained increasing attention from the scientific community as part of a One Health framework. Furthermore, the need for non-invasive sampling methods with a minimal impact on wildlife has become paramount in complying with modern ethical standards and regulations, and to collect high-quality and unbiased data. We analysed the publication trends on non-invasive sampling in wildlife health and disease research and offer a comprehensive review on the different samples that can be collected non-invasively. We retrieved 272 articles spanning from 1998 to 2021, with a rapid increase in number from 2010. Thirty-nine percent of the papers were focussed on diseases, 58% on other health-related topics, and 3% on both. Stress and other physiological parameters were the most addressed research topics, followed by viruses, helminths, and bacterial infections. Terrestrial mammals accounted for 75% of all publications, and faeces were the most widely used sample. Our review of the sampling materials and collection methods highlights that, although the use of some types of samples for specific applications is now consolidated, others are perhaps still underutilised and new technologies may offer future opportunities for an even wider use of non-invasively collected samples.

Daily torpor is a means of saving energy by controlled lowering of the metabolic rate (MR) during resting, usually coupled with a decrease in body temperature. We studied nocturnal daily torpor under natural conditions in free-living common swifts Apus apus resting in their nests as a family using two non-invasive approaches. First, we monitored nest temperature ( T nest ) in up to 50 occupied nests per breeding season in 2010-2015. Drops in T nest were the first indication of torpor. Among 16 673 observations, we detected 423 events of substantial drops in T nest of on average 8.6°C. Second, we measured MR of the families inside nest-boxes prepared for calorimetric measurements during cold periods in the breeding seasons of 2017 and 2018. We measured oxygen consumption and carbon dioxide production using a mobile indirect respirometer and calculated the percentage reduction in MR. During six torpor events observed, MR was gradually reduced by on average 56% from the reference value followed by a decrease in T nest of on average 7.6°C. By contrast, MR only decreased by about 33% on nights without torpor. Our field data gave an indication of daily torpor, which is used as a strategy for energy saving in free-living common swifts.

The identification of film support material is of utmost importance for evidence-based collection management in cultural heritage institutions, especially the identification of cellulose nitrate for fire safety reasons, as nitrate is highly flammable and deteriorates over time. Cellulose nitrate film was used by photographers and movie filmmakers from its release in the 1880s to the 1950s. Cellulose acetate, being called safety film, gradually began to replace cellulose nitrate, as it is not flammable. Despite its non-flammable properties, cellulose acetate also deteriorates in hazardous ways. Therefore, identification of cellulose nitrate and cellulose acetate in collections is imperative for preservation and risk management to collections and humans. Large photographic collections can easily contain several thousand negatives or more, so a rapid, non-invasive and reliable method is needed. Traditional identification methods, such as destructive chemical tests, are sometimes unreliable, and spectroscopic analyses are normally time-consuming. To overcome these issues, rapid material characterization was performed in transflection mode with a Fourier-transform infrared (FTIR) spectrometer equipped with an external reflectance module and an additional aluminum-foil reflector. With this newly developed method, the support material (cellulose nitrate, cellulose acetate and polyester) of about 99.8% of all films can be determined within two seconds of measuring time, without any further spectral processing. Very distinctive spectral patterns are obtained with this new method, regardless of which side of the film is being analyzed. A simple visual inspection of the raw spectrum is usually sufficient to determine the film support identity. A detailed comparison of the various FTIR techniques shows the advantages of the transflection measurement for the material characterization of film support layers. This newly developed method enables the non-invasive, rapid and unambiguous material identification of even large film collections in a short time.

With over a quarter of the world's bats species facing extinction, there is a need for ecotoxicological studies to assess if acute and sublethal exposure to newer pesticides such as neonicotinoids and carbonates contribute to population declines. Pesticide exposure studies in bats have been limited to terminal sampling methods, therefore we developed a non-invasive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method utilizing hair trimmings. The hair of big brown bats ( Eptesicus fuscus ) was collected and pooled by county to assess the best extraction solvent and solid-phase-extraction (SPE) clean-up cartridges. Using the best performing extraction solvent, methanol, and the best performing SPE cartridge, Chromabond HR-X, we developed an optimized multiple reaction monitoring (MRM) LC-MS/MS method for simultaneous determination of 3 neonicotinoids, clothianidin, imidacloprid, and thiamethoxam; 1 carbonate, carbaryl; and 4 systemic herbicides, 2,4-D, atrazine, dicamba, and glyphosate. The optimized protocol yielded the detection of 3-8 of the compounds in the county-level bat hair pools. 2,4-D, glyphosate, and imidacloprid were found in all samples with two of the county-level hair samples having glyphosate concentrations of over 3500 pg/mg of hair. This approach has great potential to facilitate non-terminal ecotoxicological studies assessing the effects of subacute (chronic) pesticide exposure in threatened and endangered bat species and other species experiencing population declines.

The human eye is a complex biomechanical structure with a range of biomechanical processes involved in various physiological as well as pathological conditions. Fluid flow inside different domains of the eye is one of the most significant biomechanical processes that tend to perform a wide variety of functions and when combined with other biophysical processes play a crucial role in ocular drug delivery. However, it is quite difficult to comprehend the effect of these processes on drug transport and associated treatment experimentally because of ethical constraints and economic feasibility. Computational modeling on the other hand is an excellent means to understand the associated complexity between these aforementioned processes and drug delivery. A wide range of computational models specific to different types of fluids present in different domains of the eye as well as varying drug delivery modes has been established to understand the fluid flow behavior and drug transport phenomenon in an insilico manner. These computational models have been used as a non-invasive tool to aid ophthalmologists in identifying the challenges associated with a particular drug delivery mode while treating particular eye diseases and to advance the understanding of the biomechanical behavior of the eye. In this regard, the author attempts to summarize the existing computational and mathematical approaches proposed in the last two decades for understanding the fluid mechanics and drug transport associated with different domains of the eye, together with their application to modify the existing treatment processes., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Epilepsy is a debilitating condition, primarily refractory individuals, leading to the search for new efficient therapies. Electrical stimulation is an important method used for years to treat several neurological disorders. Currently, electrical stimulation is used to reduce epileptic crisis in patients and shows promising results. Even though the use of electricity to treat neurological disorders has grown worldwide, there are still many caveats that must be clarified, such as action mechanisms and more efficient stimulation treatment parameters. Thus, this review aimed to explore the comprehension of the main stimulation methods in animal models of epilepsy using rodents to develop new experimental protocols and therapeutic approaches., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background and Aims: Liver stiffness measurements (LSMs) by 2-dimensional-shear-wave elastography (LSM 2D-SWE ) are now widely used in hepatology. However, relevant information for primary biliary cholangitis (PBC) is scant. We compare LSM 2D-SWE with liver biopsy (LB) in a cohort of PBC patients in Greece., Methods: Data of 68 LBs from 53 PBC patients were retrospectively analyzed and fibrosis stage was compared to LSM 2D-SWE . Forty-six patients (86.8%) were females and at the time of LBx median (IQR) age was 62.6 (53.2-72.1). Demographic, UDCA treatment, histological and B-mode ultrasound data were tested for their influence on LSM 2D-SWE estimates., Results: Liver fibrosis stages F0-F4 were found in 4, 19, 19, 16 and 10 cases, respectively. Across stages F0-F4, the LSM 2D-SWE was 5.6 (5.1-6.1), 7.0 (5.8-7.7), 9.1 (7.3-11.5), 10.8 (9.9-12.2) and 14.5 (11.9-25.7) kPa, respectively, with highly significant difference ( p <.001). The LSM 2D-SWE differed also significantly between F0 vs. F1 ( p =.027), F1 vs. F2 ( p =.005) and F3 vs. F4 ( p =.017). The discriminatory ability of LSM 2D-SWE for mild, significant, severe fibrosis and cirrhosis was highly significant in all comparisons ( p <.001), with AUC 2D-SWE 95.3%, 87.4%, 85.3% and 95.3% and accuracy 89.7%, 85.3%, 80.9% and 86.8%, respectively. Among 21 parameters tested, significant predictors of LSM 2D-SWE by multiple linear regression were fibrosis stage, portal inflammation and parenchymal heterogeneity. The portal inflammation grade accounted for 32.2% of LSM variation with adjusted R 2 0.428., Conclusions: In patients with PBC, LSM measurements by 2D-SWE can reliably discriminate between mild, significant, severe fibrosis and cirrhosis. Measurements are significantly affected by portal inflammation grade.

During operational activities, military personnel face extremely demanding circumstances, which when combined lead to severe fatigue, influencing both their well-being and performance. Physical exertion is the main condition leading to fatigue, and its continuous tracking would help prevent its effects. This review aimed to investigate the up-to-date progress on non-invasive physiological monitoring to evaluate situations of physical exertion as a pre-condition to fatigue in military populations, and determine the potential associations between physiological responses and fatigue, which can later result in decision-making indicators to prevent health-related consequences. Adhering to the PRISMA Statement, four databases (Scopus, Science Direct, Web of Science and PubMed) were used for a literature search based on combinations of keywords. The eligibility criteria focused on studies monitoring physiological variables through non-invasive objective measurements, with these measurements being developed in military field, combat, or training conditions. The review process led to the inclusion of 20 studies. The findings established the importance of multivariable assessments in a real-life context to accurately characterise the effects of military practices. A tendency for examining heart rate variables, thermal responses, and actigraphy measurements was also identified. The objectives and experimental protocols were diverse, but the effectiveness of non-invasive measurements in identifying the most fatigue-inducing periods was demonstrated. Nevertheless, no assessment system for standardised application was presented. Future work may include the development of assessment methods to translate physiological recordings into actionable information in real-time and mitigate the effects of fatigue on soldiers' performance accurately.

We aimed to investigate the diagnostic performance of new 2-D shear-wave elastography (SWE) with propagation maps and attenuation imaging (ATI) for quantification of fibrosis and steatosis in non-alcoholic fatty liver disease (NAFLD). Consecutive patients with NAFLD and healthy volunteers underwent liver stiffness measurement and steatosis quantification by means of vibration-controlled transient elastography coupled with the controlled attenuation parameter as the reference and by 2-D shear-wave elastography (2-D-SWE) with propagation maps and ATI as the investigational methods. We included 232 participants (164 in the NAFLD group and 68 in the healthy control group): 51.7%/49.3% women/men; mean age, 54.2 ± 15.2 y; mean body mass index, 29.4 ± 6.5 kg/m 2 . Significant correlations were found between 2-D-SWE and vibration-controlled transient elastography (r = 0.71, p < 0.0001) and between ATI and the controlled attenuation parameter (r = 0.72, p < 0.0001). NAFLD-specific 2-D-SWE liver stiffness measurement cutoffs were as follows-F ≥ 2: 7.9 kPa (area under the curve [AUC] = 0.91); F ≥ 3: 10 kPa (AUC = 0.92); and F = 4: 11.4 kPa (AUC = 0.95). For steatosis, the best cutoffs by ATI were as follows-S1 = 0.73 dB/cm/MHz (AUC = 0.86); S2 = 0.76 dB/cm/MHz (AUC = 0.86); and S3 = 0.80 dB/cm/MHz (AUC = 0.83). According to Baveno VI criteria, the optimal 2-D-SWE liver stiffness measurement for diagnosing liver cirrhosis is 15.5 kPa (AUC = 0.94), and for ruling out compensated advanced chronic liver disease it is 9.2 kPa (AUC = 0.92). To conclude, 2-D-SWE with propagation maps and ATI is reliable for quantification of liver fibrosis and steatosis in patients with NAFLD., (Copyright © 2021 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Background: Helicobacter pylori detection in asymptomatic children with suspected infection or with symptoms that suggest gastric pathology is problematic, since most of the methods depend on the endoscopic study, an invasive and expensive method. Non-invasive methods can be a feasible alternative but must be validated. The purpose of this study was to evaluate the concordance between H. pylori DNA detection in saliva and dental plaque by PCR, with antigen detection in stool by immunochromatography, among asymptomatic children in the state of Guerrero, Mexico., Methods: Dental plaque, saliva, and stool samples were obtained from 171 children between 6 and 12 years old. H. pylori detection in saliva and dental plaque was performed by PCR using specific primers for the 16S rRNA gene, while the detection in stool samples was performed by immunochromatography using the CerTest kit., Results: We found an overall H. pylori prevalence of 59.6% (102/171). Of the H. pylori positive children 18% (20/111) were positive in saliva samples, 28.1% (34/121) in dental plaque samples, and 50.4% (71/141) in stool samples. A higher prevalence was found in girls (64.7%, p = 0.002). Although some of the children declared some dyspeptic symptoms, these were no related to H. pylori . In conclusion, we found a high prevalence of H. pylori in asymptomatic children and the highest proportion was detected by stool antigen test, which was the most feasible method to detect H. pylori infection., Competing Interests: The authors declare that they have no competing interests., (© 2021 Martínez-Santos et al.)

Hepatitis C virus (HCV) infection and unhealthy alcohol use are major drivers of the burden of liver disease worldwide and commonly co-occur. Assessment of underlying liver damage is a cornerstone of the clinical care of patients with chronic HCV infection and/or unhealthy alcohol use because many of them are diagnosed at advanced stages of disease. Early diagnosis of liver disease before decompensated liver cirrhosis becomes established is essential for treatment with direct acting antivirals and/or abstinence from alcohol consumption, which are the main therapeutic approaches for clinical management. In this review, we discuss current knowledge around the use of non-invasive methods to assess liver disease, such as abdominal ultrasound, controlled attenuation parameter, transient elastography, magnetic resonance imaging, and indices based on serum markers of liver injury., Competing Interests: Conflict-of-interest statement: No conflict declared., (©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.)

Introduction: Human in vivo models of skin damage were often used in research of cutaneous disorders. The most commonly used models were tape-stripping as mechanical, sodium lauryl sulphate-induced irritation as chemical and ultraviolet radiation as physical damage model. In regard to differences between models, they were expected to have different responses to damage and recovery, with unique skin parameters' changes over time., Objective: The aim was to compare skin parameters in three different skin damage models on the same anatomical location, with and without topical treatment., Methods: Four test sites on each forearm were randomly assigned to three skin damage models with the fourth sites on each forearm chosen as a control, undamaged site. Skin parameters were assessed using non-invasive methods., Results: Sodium lauryl sulphate irritation caused the strongest damage with delayed reaction to the irritant. Tape stripping leads to highest initial skin barrier disruption but afterwards it showed the fastest skin recovery. Ultraviolet radiation did not affect skin barrier function, but it elevated skin erythema and melanin level. Tested preparation did not lead to changes in measured parameters., Conclusion: The skin of the participants had different response to three skin damage models with distinct changes of skin parameters and recovery. The trial was registered at ClinicalTrials.gov under the identifier NCT03783819., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: The use of non-destructive methods with less human interference is of great interest in agricultural industry and crop breeding. Modern imaging technologies enable the automatic visualization of multi-parameter for characterization of biological samples, reducing subjectivity and optimizing the analysis process. Furthermore, the combination of two or more imaging techniques has contributed to discovering new physicochemical tools and interpreting datasets in real time., Results: We present a new method for automatic characterization of seed quality based on the combination of multispectral and X-ray imaging technologies. We proposed an approach using X-ray images to investigate internal tissues because seed surface profile can be negatively affected, but without reaching important internal regions of seeds. An oilseed plant (Jatropha curcas) was used as a model species, which also serves as a multi-purposed crop of economic importance worldwide. Our studies included the application of a normalized canonical discriminant analyses (nCDA) algorithm as a supervised transformation building method to obtain spatial and spectral patterns on different seedlots. We developed classification models using reflectance data and X-ray classes based on linear discriminant analysis (LDA). The classification models, individually or combined, showed high accuracy (> 0.96) using reflectance at 940 nm and X-ray data to predict quality traits such as normal seedlings, abnormal seedlings and dead seeds., Conclusions: Multispectral and X-ray imaging have a strong relationship with seed physiological performance. Reflectance at 940 nm and X-ray data can efficiently predict seed quality attributes. These techniques can be alternative methods for rapid, efficient, sustainable and non-destructive characterization of seed quality in the future, overcoming the intrinsic subjectivity of the conventional seed quality analysis.

Lichens survive harsh weather of Antarctica as well as of other hostile environments worldwide. Therefore, this investigation is important to understand the evolution of life on Earth in relation to their stress tolerance strategy. We have used chlorophyll a fluorescence (ChlF) and Raman spectroscopy, respectively, to monitor the activation/deactivation of photosynthesis and carotenoids in three diverse Antarctic lichens, Dermatocarpon polyphyllizum (DP), Umbilicaria antarctica (UA), and Leptogium puberulum (LP). These lichens, post 4 h or 24 h of hydration, showed differences in their ChlF transients and values of major ChlF parameters, e.g., in the maximum quantum efficiency of PSII photochemistry (F v /F m ), and yields of fluorescence and heat dissipation (Φ f,d ), of effective quantum efficiency of PSII photochemistry (Φ PSII ) and of non-photochemical quenching (Φ npq ), which may be due to quantitative and/or qualitative differences in the composition of their photobionts. For understanding the kinetics of hydration-induced activation of photosynthesis, we screened Φ PSII of these lichens and reported its non-linear stimulation on a minute time scale; half of the activation time (t 1/2 ) was fastest ~4.05 ± 0.29 min for DP, which was followed by 5.46 ± 0.18 min for UA, and 13.95 ± 1.24 min for LP. Upon drying of fully activated lichen thallus, there was a slow decay, in hours, of relative water content (RWC) as well as of F v /F m . Raman spectral signatures were different for lichens having algal (in DP and UA) and cyanobacteria (in LP) photobionts, and there was a significant shift in ν 1 (C=C) Raman band of carotenoids post 24 h hydration as compared to their value at a dry state or post 4 h of hydration; this shift was decreased, when drying, in DP and LP but not in UA. We conclude that hydration nonlinearly activated photosynthetic apparatus/reactions of these lichens in minute time range but there was a de-novo synthesis of chlorophylls as well as of carotenoids post 24 h. Their dehydration-induced deactivation, however, was comparatively slow, in hours range, and there seemed a degradation of synthesized chlorophylls and carotenoids post dryness. We conclude that in extremophilic lichens, their photosynthetic partners, in particular, possess a complex survival and photoprotective strategy to be successful in the extreme terrestrial environments in Antarctica., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Chronic venous disease (CVD) is secondary to venous hypertension, leading to vascular inflammation and tissue changes. The impact of CVD on skin structure and barrier function is not well characterized., Objective: We aimed to assess the characteristics of skin alterations in mild-to-moderate CVD by non-invasive techniques based on a prospective exploratory study., Material & Methods: Female subjects (30-75 years) with CVD (Stage C2 to C4, CEAP classification) were eligible. Stage C0-C1 CVD subjects were used as controls. Women with leg surgery or a medical history that could impact the results were excluded. The skin changes on lesional (LS) and non-lesional (NLS) areas were assessed by biometric analysis including skin echography, viscoelasticity evaluation, confocal microscopy and trans epidermal water loss (TEWL) measurements., Results: Thirty-four subjects were enrolled. Based on computation of 26 biometric parameters using Principal Component Analysis, a significant difference between LS and NLS zones, regardless of the CEAP class, was evidenced. C2-C4 subjects presented with dermal thickening suggesting oedema associated with decreased cell density, while no difference in skin viscoelasticity was observed compared to the C0-C1 control group. Epidermal structural modifications were associated with increased TEWL correlating with CVD severity., Conclusion: Skin alterations in CVD patients are detectable by non-invasive methods. These findings may help to better assess new therapeutic strategies.

Hyperspectral analysis represents a powerful technique for diagnostics of morphological and chemical information from aboveground parts of the plants, but the real potential of the method in pre-screening of phenolics in leaves is still insufficiently explored. In this study, assessment of the sensitivity and reliability of non-invasive methods of various phenolic compounds, also analyzed by HPLC in chicory plants (Cichorium intybus L.) exposed to various color light pretreatments was done. The hyperspectral records in visible and near infrared (VNIR) spectra were recorded using a handheld spectrometer and relationships between the specific hyperspectral parameters and the contents of tested phenolic compounds in chicory leaves were analyzed. Moreover, the correlations between the hyperspectral parameters and related parameters derived from the multispectral fluorescence records were assessed to compare the sensitivity of both techniques. The results indicated a relatively high correlation of anthocyanin-related parameters (ARI, mARI, mACI indices) with the content of some of tested phenolic compounds (quercetin-3-gluconuride, isorhamnetine-3-gluconuride, etc.), as well as with fluorescence ANTH index. Similar trends were observed in flavonoid parameter based on the near infra-red spectral bands (700, 760 nm), which expressed a high correlation with chlorogenic acid. On the other hand, the most frequently used flavonoid (FLAVI) indices based on UV-to-blue band reflectance showed very weak correlations with phenolic compounds, as well as with fluorescence FLAV index. The detailed analysis of the correlation between reflectance and fluorescence flavonoid parameters has shown that the parameters based on spectral reflectance are sensitive to increase of UV-absorbing compounds from low to moderate values, but, unlike the fluorescence parameter, they are not useful to recognize a further increase from middle to high or very high contents. Thus, our results outlined the possibilities, but also the limits of the use of hyperspectral analysis for rapid screening phenolic content, providing a practical evidence towards more efficient production of bioactive compounds for pharmaceutical or nutraceutical use., (Copyright © 2020. Published by Elsevier Masson SAS.)

It is often hypothesized that organisms exposed to environmental change may experience physiological stress, which could reduce individual quality and make them more susceptible to disease. Amphibians are amongst the most threatened taxa, particularly in the context of disease, but relatively few studies explore links between stress and disease in amphibian species. Here, we use the fungal pathogen Batrachochytrium dendrobatidis (Bd) and amphibians as an example to explore relationships between disease and glucocorticoids (GCs), metabolic hormones that comprise one important component of the stress response. While previous work is limited, it has largely identified positive relationships between GCs and Bd-infection. However, the causality remains unclear and few studies have integrated both baseline (GC release that is related to standard, physiological functioning) and stress-induced (GC release in response to an acute stressor) measures of GCs. Here, we examine salivary corticosterone before and after exposure to a stressor, in both field and captive settings. We present results for Bd-infected and uninfected individuals of three amphibian species with differential susceptibilities to this pathogen (Rana catesbeiana, R. clamitans, and R. sylvatica). We hypothesized that prior to stress, baseline GCs would be higher in Bd-infected animals, particularly in more Bd-susceptible species. We also expected that after exposure to a stressor, stress-induced GCs would be lower in Bd-infected animals. These species exhibited significant interspecific differences in baseline and stress induced corticosterone, though other variables like sex, body size, and day of year were usually not predictive of corticosterone. In contrast to most previous work, we found no relationships between Bd and corticosterone for two species (R. catesbeiana and R. clamitans), and in the least Bd-tolerant species (R. sylvatica) animals exhibited context-dependent differences in relationships between Bd infection and corticosterone: Bd-positive R. sylvatica had significantly lower baseline and stress-induced corticosterone, with this pattern being stronger in the field than in captivity. These results were surprising, as past work in other species has more often found elevated GCs in Bd-positive animals, a pattern that aligns with well-documented relationships between chronically high GCs, reduced individual quality, and immunosuppression. This work highlights the potential relevance of GCs to disease susceptibility in the context of amphibian declines, while underscoring the importance of characterizing these relationships in diverse contexts., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: Patients with nonalcoholic steatohepatitis (NASH) are at risk for developing cirrhosis and hepatic cancer. Currently, the definitive gold-standard method of diagnosing NASH is a liver biopsy, an invasive and costly method. Our objective was to compare three non-invasive methods of identifying NASH by using data on 10,007 subjects from NHANES III (1988-1994) to determine the prevalence and variables associated with NASH, as defined by each non-invasive method., Methods: We used ultrasound data to identify subjects with moderate-to-severe hepatic steatosis, of whom we identified the NASH population using either the HAIR score, the NASH liver fat score, or the Gholam score, each of which had been validated with liver biopsy. We performed multinomial logistic regression to compare each NASH population to the normal population (those with no-to-mild hepatic steatosis)., Results: We identified 1136 (9.5%) subjects as having NASH by at least one method and 219 (1.8%) were identified by all 3 methods. Independent of the non-invasive method used, Mexican-Americans (MA) had the highest prevalence of NASH. All three methods identified significant risk factors for NASH (p<0.05), including: elevated waist-to-hip ratio, elevated levels of C-peptide, total cholesterol, or C-reactive protein (CRP)., Conclusion: We conclude that the combined non-invasive methods can help identify candidates with a high likelihood of being diagnosed with NASH. Health care providers can screen people with the combined non-invasive methods for the risk factors and identify candidates for interventions, including exercise and/or referral to biopsy.

Background & Objectives: The non-invasive method of haemoglobin (Hb) estimation has unique advantages of exemption of finger prick and associated pain, over invasive methods. This study was done to compare invasive and non-invasive methods of Hb estimation in blood donors keeping haematology analyzer (HA) as a reference method., Methods: The blood donors selected or deferred on the basis of CuSO 4 method (Hb ≥12.5 g/dl), were included in the study. Hb values of the donors were estimated by HemoCue and then by OrSense methods. An immediate post-donation venous sample was drawn for analysis on HA., Results: The mean Hb value was 13.98±1.27 g/dl on HA, 14.87±1.03 g/dl on OrSense and 15.03±1.31 g/dl on HemoCue. CuSO 4 , HemoCue and OrSense demonstrated sensitivities of 18.7, 18.7 and 13.1 per cent, positive predictive values (PPV) of 64.5, 83.3 and 60.9 per cent and specificities of 98.9, 99.6 and 99.1 per cent, respectively. The intra-class correlation coefficient for OrSense was 0.726 while that for HemoCue was 0.851. Bland-Altman plots demonstrated 2SD difference of >2.0 g/dl in Hb estimations between HA and HemoCue/OrSense., Interpretation & Conclusions: The non-invasive modality may provide the near-ideal pre-donation Hb screening platform if an improvement can be done in the sensitivity and PPV of the non-invasive method keeping in view its unique advantages., Competing Interests: None

Structural analysis of marble statues, carried out by non-invasive and in situ methods, is crucial to define the state of conservation of the artworks and to identify the deterioration phenomena that can affect them. In this work, we combined in situ non-destructive techniques, ultrasonic tomography (US), ultraviolet-induced visible fluorescence (UV-IF) and X-ray fluorescence (XRF) to study the bass-relief 'Madonna con Bambino' (Gorizia, Italy). The US revealed the presence of some metallic pivots, associated with areas of high sound velocity; moreover, a more degraded area has been identified in the lower part of the bass-relief. The acquired UV-IF image confirmed the presence of surface degradation, allowing a preliminary evaluation of the extension of a fracture, from surface to bulk. In addition, the different materials (both original and/or integrations) that compose the studied surface have been identified. The XRF has contributed to define the nature of the inorganic materials applied during undocumented previous restoration works on the surface as filler for lacunae.

The study was performed at Brasov County Hospital, in the Internal Medicine, Diabetes, Gastroenterology and Cardiology Wards with the collaboration of Transylvania University of Brasov, as a study approved by the Ethical Board of the university. The study aimed at assessing the connection between the anthropometric parameters of abdominal adiposity (measured by means of an original experiment designed to determine the curvature of the thoracic-abdominal adiposity for the patients and processed by help of a mathematical model based upon Bezier curves geometry) and the fat load of the liver (assessed by ultrasound by measuring the diameters of both hepatic lobes) for the patients diagnosed with Non-Alcoholic Fatty Liver Disease (NAFLD). The existence of ten types of thoracic-abdominal curves profiles were statistically analyzed in order to evaluate in a simple manner the liver size in NAFLD. The method of diagnosis is based on an easily reproduced experiment, it is original, innovative, non-invasive, and cost-effective. Can be implemented anywhere in the world, there is no need for investment, only for determining the profile of the belly.

Background and Aims: Non-invasive fibrosis assessment has been highly recommended in many liver diseases. However, comparative diagnostic accuracy of laboratory markers, ultrasound and magnetic resonance elastography (MRE) for fibrosis in autoimmune hepatitis (AIH) patients has not been established., Methods: Medline, Embase and Cochrane Library were searched. Primary outcome was significant fibrosis (SF), advanced fibrosis (AF) and cirrhosis, defined as Metavir stage F ≥ 2, F ≥ 3 and F = 4 according to liver biopsy. Hierarchical summary receiver operating characteristic curve (ROC) model was used to evaluate diagnostic accuracy of non-invasive methods. Summary area under ROC (AUROC) and diagnostic odds ratio (DOR) with 95% confidence interval (CI) were calculated. The Grading of Recommendations Assessment, Development and Evaluation system was used to assess quality of evidence., Results: Overall, 16 studies with 861 patients were included, comparing aspartate aminotransferase to platelet ratio index (APRI), fibrosis-4 index (FIB-4), aspartate aminotransferase/alanine aminotransferase ratio, transient elastography (TE), acoustic radiation force impulse, shear wave elastography and MRE versus liver biopsy. Among all non-invasive markers, TE had good performance for fibrosis staging. Summary AUROCs and DORs of TE were 0.90 (95% CI 0.87, 0.92) and 23.7, 0.91 (95% CI 0.89, 0.93) and 31.6, 0.89 (95% CI 0.86, 0.92) and 80.5 for staging SF, AF and cirrhosis, whereas APRI and FIB-4 showed poor performance for detecting AF (DOR, 4.6 and 4.7) and cirrhosis (DOR, 5.5 and 12.9)., Conclusions: TE performs well to stage liver fibrosis in patients with AIH, compared with other laboratory non-invasive indexes. Nevertheless, diagnostic accuracy of APRI and FIB-4 is poor.

Conventionally, plasma or milk progesterone evaluations are used to determine the reproductive status of female animals. Collection of such samples is often associated with difficulties of animal handling and restraint. Measurable quantities of progesterone metabolites are found in feces of animals. Their concentrations are known to be well correlated to plasma progesterone levels and are, therefore, used as non-invasive samples for assessing reproductive function in a wide range of animal species. Although the analysis of fecal progesterone metabolites has been widely accepted in many laboratories, several factors are known to affect the results from this valuable analytical technique. Some of these factors include storage/transportation media for fecal samples, type of solvent that is used for extraction of progesterone metabolites from feces, and the type and sensitivity of an assaying technique employed. Although fecal progesterone metabolites analysis is associated with some difficulties, it can effectively be used to monitor reproductive function in a wide range of animal species. This review aims to highlight the usefulness of fecal progesterone metabolite analysis as a non-invasive technique in monitoring reproductive function in animals. The article mainly focuses on the many opportunities and challenges associated with this analytical technique.

Hepatitis C virus (HCV) infection remains an important global health problem with chronic infection affecting approximately 11 million children worldwide. The emergence of direct-acting antiviral (DAA) therapies and the development of non-invasive methods for the determination of liver fibrosis will significantly improve the management of paediatric patients with chronic HCV infection in subsequent years. For paediatric patients, a new era of highly effective DAA agents is beginning, and the first results of available clinical trials are very promising. In this era, the identification and monitoring of patients continues to be an important issue. The availability of non-invasive serological and imaging methods to measure hepatic fibrosis enables the identification of patients with significant or advanced liver fibrosis stages. This article summarizes the current data on the epidemiology and progress of research aimed to evaluate the new therapies and non-invasive methods for liver injury in paediatric patients with chronic hepatitis C., Competing Interests: Conflict-of-interest statement: The authors declare that they have no conflicts of interest.

Background: As a landscape architect and a major seed disperser, the lowland tapir ( Tapirus terrestris ) is an important indicator of the ecological health of certain habitats. Therefore, reliable data regarding tapir populations are fundamental in understanding ecosystem dynamics, including those associated with the Atlantic Forest in Brazil. Currently, many population monitoring studies use invasive tagging with radio or satellite/Global Positioning System (GPS) collars. These techniques can be costly and unreliable, and the immobilization required carries physiological risks that are undesirable particularly for threatened and elusive species such as the lowland tapir., Methods: We collected data from one of the last regions with a viable population of lowland tapir in the south-eastern Atlantic Forest, Brazil, using a new non-invasive method for identifying species, the footprint identification technique (FIT)., Results: We identified the minimum number of tapirs in the study area and, in addition, we observed that they have overlapping ranges. Four hundred and forty footprints from 46 trails collected from six locations in the study area in a landscape known to contain tapir were analyzed, and 29 individuals were identified from these footprints., Discussion: We demonstrate a practical application of FIT for lowland tapir censusing. Our study shows that FIT is an effective method for the identification of individuals of a threatened species, even when they lack visible natural markings on their bodies. FIT offers several benefits over other methods, especially for tapir management. As a non-invasive method, it can be used to census or monitor species, giving rapid feedback to managers of protected areas., Competing Interests: The authors declare that they have no competing interests.

Objective: Detecting significant fibrosis and cirrhosis remains important in treatment and follow-up of patients with chronic hepatitis C Infection (CHC). The aim of this study was to assess the ability of PRO-C3 to identify significant fibrosis (Ishak score ≥3) and cirrhosis (Ishak score ≥5) both as a single test and as a part of algorithms., Materials and Methods: PRO-C3 was assessed in baseline samples from the NORDynamIC trial. 270 patients were stratified into groups according to baseline biopsy. Baseline APRI, FIB-4 and GUCI scores were available for comparison in 232 patients., Results: PRO-C3 increased with Ishak scores (p = .001). Area under the curve (AUC) for significant fibrosis was 0.75 (95% CI 0.68-0.81) and 0.76 (95% CI 0.68-0.84) for cirrhosis. FIB-4, APRI and GUCI had similar AUCs. In a PRO-C3 algorithm including age, platelet count, body mass index (BMI) and international normalised ratio (INR), the diagnostic efficacy improved to 0.85 (CI 0.80-0.89) and 0.90 (IQR 0.84-0.96) for significant fibrosis and cirrhosis, respectively., Conclusions: In our study, PRO-C3 was an independent predictor of fibrosis stage, and may play an important role in managing CHC patients.

Background: Liver cirrhosis is the fourth cause of death in adults in Western countries, with complications of portal hypertension being responsible for most casualties. In order to reduce mortality, development of accurate diagnostic methods for early diagnosis, effective etiologic treatment, improved pharmacological therapy for portal hypertension, and effective therapies for end-stage liver failure are required., Discussion: Early detection of cirrhosis and portal hypertension is now possible using simple non-invasive methods, leading to the advancement of individualized risk stratification in clinical practice. Despite previous assumptions, cirrhosis can regress if its etiologic cause is effectively removed. Nevertheless, while this is now possible for cirrhosis caused by chronic hepatitis C, the incidence of cirrhosis due to non-alcoholic steatohepatitis has increased dramatically and effective therapies are not yet available. New drugs acting on the dynamic component of hepatic vascular resistance are being studied and will likely improve the future management of portal hypertension., Conclusion: Cirrhosis is now seen as a dynamic disease able to progress and regress between the compensated and decompensated stages. This opinion article aims to provide the author's personal view of the current major advances and challenges in this field.

The bank vole is a commonly used model species in behavioral and ecophysiological studies. Thus, presenting a validated method for noninvasive monitoring of corticosterone and testosterone secretion is of high relevance. Here, we evaluated the effect of time of day and an ACTH challenge test on measured fecal corticosterone (FCM) and testosterone (FTM) metabolites in both sexes. Furthermore, we performed radiometabolism experiments for both steroids and sexes to study metabolism and excretion of 3 H-corticosterone and 3 H-testosterone. FCM and FTM were analysed with a 5α-pregnane-3β,11β,21-triol-20-one enzyme immunoassay (EIA) and a testosterone (measuring 17β-hydroxyandrostanes) EIA, respectively. Males had significantly higher FCM levels than females and their main excretion route was via the feces (∼72%), whereas females excreted nearly equal portions in both feces and urine. For testosterone the main excretion route was via the feces in both sexes (∼80%). The time course of excretion was similar in both sexes, but for the first time a significant difference between injected steroids was found: Corticosterone was excreted faster than testosterone, both in urine (median of peak levels: 4h vs 6h) and feces (6h vs 8h). Several metabolites were present in the feces and the tested EIAs reacted with some of them. Time of day had a significant effect on measured fecal steroid metabolites. As expected, males had significantly higher FTM levels than females. ACTH administration significantly increased FCM values; peaks were observed 4-8h after injection. In conclusion, both tested EIAs proved suited for a noninvasive measurement of glucocorticoids and androgens in bank voles., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Aim: To evaluate the performance of FibroMeter Virus3G combined to the first generation tests aspartate aminotransferase-to-platelet ratio index (APRI) or Forns index to assess significant fibrosis in chronic hepatitis C (CHC)., Methods: First generation tests APRI or Forns were initially applied in a derivation population from Rio de Janeiro in Brazil considering cut-offs previously reported in the literature to evaluate significant fibrosis. FibroMeter Virus3G was sequentially applied to unclassified cases from APRI or Forns. Accuracy of non-invasive combination of tests, APRI plus FibroMeter Virus3G and Forns plus FibroMeter Virus3G was evaluated in the Brazilian derivation population. APRI plus FibroMeter Virus3G combination was validated in a population of CHC patients from Angers in France. All patients were submitted to liver biopsy staged according to METAVIR score by experienced hepatopathologists. Significant fibrosis was considered as METAVIR F ≥ 2. The fibrosis stage classification was used as the reference for accuracy evaluation of non-invasive combination of tests. Blood samples for the calculation of serum tests were collected on the same day of biopsy procedure or within a maximum 3 mo interval and stored at -70 °C., Results: Seven hundred and sixty CHC patients were included (222 in the derivation population and 538 in the validation group). In the derivation population, the FibroMeter Virus3G AUROC was similar to APRI AUROC (0.855 vs 0.815, P = 0.06) but higher than Forns AUROC (0.769, P < 0.001). The best FibroMeter Virus3G cut-off to discriminate significant fibrosis was 0.61 (80% diagnostic accuracy; 75% in the validation population, P = 0.134). The sequential combination of APRI or Forns with FibroMeter Virus3G in derivation population presented similar performance compared to FibroMeter Virus3G used alone (79% vs 78% vs 80%, respectively, P = 0.791). Unclassified cases of significant fibrosis after applying APRI and Forns corresponded to 49% and 54%, respectively, of the total sample. However, the combination of APRI or Forns with FibroMeter Virus3G allowed 73% and 77%, respectively, of these unclassified cases to be correctly evaluated. Moreover, this combination resulted in a reduction of FibroMeter Virus3G requirement in approximately 50% of the entire sample. The stepwise combination of APRI and FibroMeter Virus3G applied to the validation population correctly identified 74% of patients with severe fibrosis (F ≥ 3)., Conclusion: The stepwise combination of APRI or Forns with FibroMeter Virus3G may represent an accurate lower cost alternative when evaluating significant fibrosis, with no need for liver biopsy., Competing Interests: Conflict-of-interest statement: No potential conflicts of interest relevant to this article were reported.