Multispectral Portable Fibre-Optic Reflectometer for the Classification of the Origin of Chicken Eggshells in the Case of Mycoplasma synoviae Infections.

The proper classification of the origins of food products is a crucial issue all over the world nowadays. In this paper, the authors present a device-a multispectral portable fibre-optic reflectometer and signal processing patch-together with a machine-learning algorithm for the classification of the origins of chicken eggshells in the case of Mycoplasma synoviae infection. The sensor device was developed based on previous studies with a continuous spectrum in transmittance and selected spectral lines in reflectance. In the described case, the sensor is based on the integration of reflected spectral data from short spectral bands from the VIS and NIR region, which are produced by single-colour LEDs and introduced to the sample via a fibre bundle. The measurement is carried out in a sequence, and the reflected signal is pre-processed to be put in the machine learning algorithm. The support vector machine algorithm is used together with three different types of data normalization. The obtained results of the F-score factor for classification of the origins of samples show that the percentages of eggs coming from Mycoplasma synoviae infected hens are up to 87% for white and 96% for brown eggshells.

Recognized and Potentially New Biomarkers-Their Role in Diagnosis and Prognosis of Cardiovascular Disease.

Atherosclerosis and its consequences are the leading cause of mortality in the world. For this reason, we have reviewed atherosclerosis biomarkers and selected the most promising ones for review. We focused mainly on biomarkers related to inflammation and oxidative stress, such as the highly sensitive C-reactive protein (hs-CRP), interleukin 6 (IL-6), and lipoprotein-associated phospholipase A2 (Lp-PLA2). The microRNA (miRNA) and the usefulness of the bone mineralization, glucose, and lipid metabolism marker osteocalcin (OC) were also reviewed. The last biomarker we considered was angiogenin (ANG). Our review shows that due to the multifactorial nature of atherosclerosis, no single marker is known so far, the determination of which would unambiguously assess the severity of atherosclerosis and help without any doubt in the prognosis of cardiovascular risk.

An attempt to classify the botanical origin of honey using visible spectroscopy.

BACKGROUND: The production of honey, and especially the unifloral varieties, is limited by factors such as weather conditions or the availability of nectar flow and honeydew. This results in a deficit in supply leading to the adulteration of honey. If they are not properly labeled, customers cannot distinguish artificial / synthetic products from real honey. Currently, the basic, commonly used method for determining the varieties of honey (botanical origin) is palynological analysis. However, this procedure is quite difficult owing to the dearth of experienced staff in the field of melissopalynology. RESULTS: A method for identifying and classifying natural honey accurately based on its botanical origin has therefore been proposed. This analysis would rely on the visible light spectra transmitted through a relatively thin layer of the substance of interest, regardless of deviations in thickness. We present algorithms for analyzing the transmittance spectra-parametrization based on polynomial approximation (PMA) and applying a method for spectra selection and reduction (SSR) and a classical classification model (decision tree). A comparison is presented of the classification of four varieties of honey, confirmed by pollen analysis, obtained from the analysis of optically measured transmittance spectra of the samples. The algorithms that are compared contain a decision tree that uses raw data, data reduced by principal component analysis (PCA), and data after calculations based on the proposed algorithms alone (PMA and SSR) and together with the PCA method. CONCLUSION: This novel method produced outstanding results in comparison with the standard PCA method and is helpful in identifying the botanical origin of honey effectively. © 2021 Society of Chemical Industry.

PDE10A Inhibition Reduces the Manifestation of Pathology in DMD Zebrafish and Represses the Genetic Modifier PITPNA.

Duchenne muscular dystrophy (DMD) is a severe genetic disorder caused by mutations in the DMD gene. Absence of dystrophin protein leads to progressive degradation of skeletal and cardiac function and leads to premature death. Over the years, zebrafish have been increasingly used for studying DMD and are a powerful tool for drug discovery and therapeutic development. In our study, a birefringence screening assay led to identification of phosphodiesterase 10A (PDE10A) inhibitors that reduced the manifestation of dystrophic muscle phenotype in dystrophin-deficient sapje-like zebrafish larvae. PDE10A has been validated as a therapeutic target by pde10a morpholino-mediated reduction in muscle pathology and improvement in locomotion, muscle, and vascular function as well as long-term survival in sapje-like larvae. PDE10A inhibition in zebrafish and DMD patient-derived myoblasts were also associated with reduction of PITPNA expression that has been previously identified as a protective genetic modifier in two exceptional dystrophin-deficient golden retriever muscular dystrophy (GRMD) dogs that escaped the dystrophic phenotype. The combination of a phenotypic assay and relevant functional assessments in the sapje-like zebrafish enhances the potential for the prospective discovery of DMD therapeutics. Indeed, our results suggest a new application for a PDE10A inhibitor as a potential DMD therapeutic to be investigated in a mouse model of DMD.

Applying genome-wide CRISPR-Cas9 screens for therapeutic discovery in facioscapulohumeral muscular dystrophy.

The emergence of CRISPR-Cas9 gene-editing technologies and genome-wide CRISPR-Cas9 libraries enables efficient unbiased genetic screening that can accelerate the process of therapeutic discovery for genetic disorders. Here, we demonstrate the utility of a genome-wide CRISPR-Cas9 loss-of-function library to identify therapeutic targets for facioscapulohumeral muscular dystrophy (FSHD), a genetically complex type of muscular dystrophy for which there is currently no treatment. In FSHD, both genetic and epigenetic changes lead to misexpression of DUX4, the FSHD causal gene that encodes the highly cytotoxic DUX4 protein. We performed a genome-wide CRISPR-Cas9 screen to identify genes whose loss-of-function conferred survival when DUX4 was expressed in muscle cells. Genes emerging from our screen illuminated a pathogenic link to the cellular hypoxia response, which was revealed to be the main driver of DUX4-induced cell death. Application of hypoxia signaling inhibitors resulted in increased DUX4 protein turnover and subsequent reduction of the cellular hypoxia response and cell death. In addition, these compounds proved successful in reducing FSHD disease biomarkers in patient myogenic lines, as well as improving structural and functional properties in two zebrafish models of FSHD. Our genome-wide perturbation of pathways affecting DUX4 expression has provided insight into key drivers of DUX4-induced pathogenesis and has identified existing compounds with potential therapeutic benefit for FSHD. Our experimental approach presents an accelerated paradigm toward mechanistic understanding and therapeutic discovery of a complex genetic disease, which may be translatable to other diseases with well-established phenotypic selection assays.

Spectral technique for detection of changes in eggshells caused by Mycoplasma synoviae.

Mycoplasma synoviae (MS) is a major pathogen in chicken and turkeys, causing subclinical infection. MS infections are highly prevalent and may potentate and be involved in sinovitis, respiratory syndromes, as well as lead to eggshell apex abnormality (EAA). A deformed, inhomogeneous eggshell is susceptible to cracks and breaks through which microbes get in and additionally entails higher water loss in the egg during the entire incubation process. Not all eggs with eggshell apex abnormality possess characteristic deformation and that is why some eggs may be incorrectly classified during a visual inspection. To minimize the above risk, the spectral VIS technique and the analysis based on the classification tree method-CTM is proposed. The method makes use of specially defined parameters extracted from the shape of transmittance spectra of eggshells. Directional coefficients of the lines adjusted to the specific ranges of the transmittance spectrum are used in the process of classifying samples as those from MS-carrying hens and from healthy hens. Three CTM-based classifiers were created for a group of white, brown, and mixed shells. After comparing, it can be concluded that the best results were obtained for the group of brown shells (accuracy 88%, specificity 88%, and false negative rate 13%). The authors present a non-invasive spectral method that utilizes eggshells, i.e., the natural waste from chicken farms. The method enables entering data into the classifiers described in the article. The process provides an opportunity to correctly assign, the examined shell to the group of shells with increased risk-with approx. 86% accuracy. This means that, if a few of such results are registered, the herd is eligible more specific studies targeting MS bacteria. Regular spectral testing can support the detection of egg lesions in MS positive flocks.

Eggshell apex abnormalities caused by two different Mycoplasma synoviae genotypes and evaluation of eggshell anomalies by full-field optical coherence tomography.

BACKGROUND: Mycoplasma synoviae (MS) is an important poultry pathogen worldwide. This bacterium may cause eggshell changes including an altered shell surface, thinning, and increased translucency in different areas, which leads to a greater incidence of eggshell cracks and breaks. In the present study the association between experimental infection of birds with two field strains of MS from different genotypes and the production of abnormal eggs is described. The analysis of those eggshells using a full-field optical coherence tomography (FF OCT) scanner is also reported. RESULTS: Eggshell samples were obtained from three experimental groups of chickens: one control and two infected tracheally with field strains of MS which produced abnormal eggs. In both experimental groups infected with MS a reduction of mean daily egg production by 11% was observed compared to the control group, which started at 21 to 42 dpi. Eggshell apex abnormalities increased to 24.5% of eggs and in some cases, soft-shelled eggs were produced. This study provides the first analysis of shells from anomalous eggs carried out using FF OCT, which allows three-dimensional structural imaging of an investigated sample at micrometre scale. FF OCT showed ultrastructural changes in eggshells and a smaller number of pores on the entire surface of the affected shells. CONCLUSIONS: The eggshell pathology and the concomitant egg production losses that result from infections highlight the economic significance of MS in commercial poultry. There are differences in the strains of MS which may induce eggshell apex abnormalities (EAA) and egg production losses. The use of FF OCT, which is a noninvasive measurement method based on analysis of the light backscattered from the measured object, will confer the ability to control the quality of eggshells in flocks infected with MS.

Transgenic zebrafish model of DUX4 misexpression reveals a developmental role in FSHD pathogenesis.

Facioscapulohumeral dystrophy type 1 (FSHD-1) is the most common autosomal dominant form of muscular dystrophy with a prevalence of ∼1 in 8000 individuals. It is considered a late-onset form of muscular dystrophy and leads to asymmetric muscle weakness in the facial, scapular, trunk and lower extremities. The prevalent hypothesis on disease pathogenesis is explained by misexpression of a germ line, primate-specific transcription factor DUX4-fl (double homeobox 4, full-length isoform) linked to the chromosome 4q35. In vitro and in vivo studies have demonstrated that very low levels of DUX4-fl expression are sufficient to induce an apoptotic and/or lethal phenotype, and therefore modeling of the disease has proved challenging. In this study, we expand upon our previously established injection model of DUX4 misexpression in zebrafish and describe a DUX4-inducible transgenic zebrafish model that better recapitulates the expression pattern and late onset phenotype characteristic of FSHD patients. We show that an induced burst of DUX4 expression during early development results in the onset of FSHD-like phenotypes in adulthood, even when DUX4 is no longer detectable. We also utilize our injection model to study long-term consequences of DUX4 expression in those that fail to show a developmental phenotype. Herein, we introduce a hypothesis that DUX4 expression during developmental stages is sufficient to induce FSHD-like phenotypes in later adulthood. Our findings point to a developmental role of DUX4 misexpression in the pathogenesis of FSHD and should be factored into the design of future therapies.

Purification of Myogenic Progenitors from Human Muscle Using Fluorescence-Activated Cell Sorting (FACS).

Primary myoblasts derived from human tissue are a valuable tool in research of muscle disease and pathophysiology. However, skeletal muscle biopsies, especially from diseased muscle, contain a plethora of non-myogenic cells, necessitating purification of the myogenic cell population. This protocol describes techniques for dissociation of cells from human skeletal muscle biopsies and enrichment for a highly myogenic population by fluorescence-activated cell sorting (FACS). We also describe methods for assessing myogenicity and population expansion for subsequent in vitro study.

An open source microcontroller based flume for evaluating swimming performance of larval, juvenile, and adult zebrafish.

Zebrafish are a preferred vertebrate model for delineating genotype-phenotype relationships. One of the most studied features of zebrafish is their exceptional swimming ability. By 7 days postfertilization (dpf), zebrafish spend over two-thirds of their time engaged in spontaneous swimming activity and several months later they are capable of attaining some of the fastest swimming velocities relative to body length ever recorded in the laboratory. However, laboratory-assembled flumes capable of achieving the slow flow velocities characteristics of larvae as well as the relatively fast maximal velocities of adults have not been described in sufficient detail to allow easy replication. Here we describe an easily assembled, open-source zebrafish-scaled flume for assessing swimming performance. The flume uses two independent spherical-impeller pumps modulated by a microcontroller to achieve flow velocities ranging from 1 to 70 cm s-1. The microcontroller also monitors water temperature and flow velocity and sends these data to a personal computer for real-time display and storage. Incremental protocols for assessing maximal swimming speed (Umax) were developed, stored in custom software, and then uploaded to the microcontroller in order to assess performance of larval (14, 21, 28 dpf), juvenile (35, 42 dpf), and adult (8, 22 month) zebrafish. The flume had sufficient range and sensitivity to detect developmental changes in Umax of larvae and juveniles, an 18-24% faster Umax of adult males vs. females, and a 14-20% age-related reduction in Umax for the oldest zebrafish. Detailed information is provided to assemble and operate this low-cost, versatile, and reliable tool for assessing zebrafish swimming performance.

Facioscapulohumeral Muscular Dystrophy.

Facioscapulohumeral Muscular Dystrophy is a common form of muscular dystrophy that presents clinically with progressive weakness of the facial, scapular, and humeral muscles, with later involvement of the trunk and lower extremities. While typically inherited as autosomal dominant, facioscapulohumeral muscular dystrophy (FSHD) has a complex genetic and epigenetic etiology that has only recently been well described. The most prevalent form of the disease, FSHD1, is associated with the contraction of the D4Z4 microsatellite repeat array located on a permissive 4qA chromosome. D4Z4 contraction allows epigenetic derepression of the array, and possibly the surrounding 4q35 region, allowing misexpression of the toxic DUX4 transcription factor encoded within the terminal D4Z4 repeat in skeletal muscles. The less common form of the disease, FSHD2, results from haploinsufficiency of the SMCHD1 gene in individuals carrying a permissive 4qA allele, also leading to the derepression of DUX4, further supporting a central role for DUX4. How DUX4 misexpression contributes to FSHD muscle pathology is a major focus of current investigation. Misexpression of other genes at the 4q35 locus, including FRG1 and FAT1, and unlinked genes, such as SMCHD1, has also been implicated as disease modifiers, leading to several competing disease models. In this review, we describe recent advances in understanding the pathophysiology of FSHD, including the application of MRI as a research and diagnostic tool, the genetic and epigenetic disruptions associated with the disease, and the molecular basis of FSHD. We discuss how these advances are leading to the emergence of new approaches to enable development of FSHD therapeutics. © 2017 American Physiological Society. Compr Physiol 7:1229-1279, 2017.

Monitoring of spine curvatures and posture during pregnancy using surface topography - case study and suggestion of method.

BACKGROUND: Low back and pelvic pain is one of the most frequently reported disorders in pregnancy, however etiology and pathology of this problem have not been fully determined. The relationship between back pain experienced during pregnancy and posture remains unclear. It is challenging to measure reliably postural and spinal changes at the time of pregnancy, since most imaging studies cannot be used due to the radiation burden. 3D shape measurement, or surface topography (ST), systems designed for posture evaluation could potentially fill this void. A pilot study was conducted to test the potential of monitoring the change of spine curvatures and posture during pregnancy using surface topography. A single case was studied to test the methodology and preliminarily assess the usefulness of the procedure before performing a randomized trial. The apparatus used in this study was metrologically tested and utilized earlier in scoliosis screening. CASE PRESENTATION: The subject was measured using a custom-made structured light illumination scanner with accuracy of 0.2 mm. Measurement was taken every 2 weeks, between 17th and 37th week of pregnancy, 11 measurements in total. From the measurement the thoracic kyphosis and lumbar lordosis angles, and vertical balance angle were extracted automatically. Custom-written software was used for analysis. Oswestry Low Back Pain Disability Questionnaire (ODI) was done with every measurement. The values were correctly extracted from the measurement. The results were: 50.9 ± 2.4° for kyphosis angle, 58.1 ± 2.1° for lordosis angle and 4.7 ± 1.7° for vertical balance angle. The registered change was 7.4° in kyphosis angle, 8.4° in lordosis angle and 5.5° in vertical balance angle. The calculated ODI values were between moderate disability and severe disability (22 to 58 %). CONCLUSIONS: This case study presents that surface topography may be suitable for monitoring of spinal curvature and posture change in pregnant women. The ionizing radiation studies are contraindicated during pregnancy. Surface topography data connected with information from pain level questionnaires allows to investigate the connection between changes in posture and back pain.

Source spectrum shaping method for low-coherence interferometry.

Low-coherence interferometry (LCI) suffers mainly from low signal-to-noise (S/N) ratio. By using the source spectrum shaping method (SSSM), the authors can enhance the visibility of the zero-order fringe (V) and S/N ratio in LCI. The presented approach was analyzed numerically for a set of theoretical Gaussian light sources, with different central wavelengths and different spectrum widths. The results have shown a significant improvement of the visibility V. Additionally, a set of commercially available light emitting diodes was analyzed to find the best possible setup. Results of numerical calculations were verified experimentally in an SSSM low coherence Twyman-Green interferometric setup equipped with three light sources.

Altered expression of cyclin A 1 in muscle of patients with facioscapulohumeral muscle dystrophy (FSHD-1).

OBJECTIVES: Cyclin A1 regulates cell cycle activity and proliferation in somatic and germ-line cells. Its expression increases in G1/S phase and reaches a maximum in G2 and M phases. Altered cyclin A1 expression might contribute to clinical symptoms in facioscapulohumeral muscular dystrophy (FSHD). METHODS: Muscle biopsies were taken from the Vastus lateralis muscle for cDNA microarray, RT-PCR, immunohistochemistry and Western blot analyses to assess RNA and protein expression of cyclin A1 in human muscle cell lines and muscle tissue. Muscle fibers diameter was calculated on cryosections to test for hypertrophy. RESULTS: cDNA microarray data showed specifically elevated cyclin A1 levels in FSHD vs. other muscular disorders such as caveolinopathy, dysferlinopathy, four and a half LIM domains protein 1 deficiency and healthy controls. Data could be confirmed with RT-PCR and Western blot analysis showing up-regulated cyclin A1 levels also at protein level. We found also clear signs of hypertrophy within the Vastus lateralis muscle in FSHD-1 patients. CONCLUSIONS: In most somatic human cell lines, cyclin A1 levels are low. Overexpression of cyclin A1 in FSHD indicates cell cycle dysregulation in FSHD and might contribute to clinical symptoms of this disease.