Spherical Aberration and Scattering Compensation in Microscopy Images through a Blind Deconvolution Method.

The optical quality of an image depends on both the optical properties of the imaging system and the physical properties of the medium the light passes while travelling from the object to the image plane. The computation of the point spread function (PSF) associated to the optical system is often used to assess the image quality. In a non-ideal optical system, the PSF is affected by aberrations that distort the final image. Moreover, in the presence of turbid media, the scattering phenomena spread the light at wide angular distributions that contribute to reduce contrast and sharpness. If the mathematical degradation operator affecting the recorded image is known, the image can be restored through deconvolution methods. In some scenarios, no (or partial) information on the PSF is available. In those cases, blind deconvolution approaches arise as useful solutions for image restoration. In this work, a new blind deconvolution method is proposed to restore images using spherical aberration (SA) and scatter-based kernel filters. The procedure was evaluated in different microscopy images. The results show the capability of the algorithm to detect both degradation coefficients (i.e., SA and scattering) and to restore images without information on the real PSF.

Surgical correction of cubitus varus in children with a lateral closing-wedge osteotomy: a comparison between two different techniques.

OBJECTIVES: The primary objective of the present study is to compare the radiographic outcomes and complications of two different techniques for lateral closing-wedge osteotomy in pediatric patients with cubitus varus. METHODS: We retrospectively identified patients treated at five tertiary care institutions: 17 underwent the Kirschner-wire (KW) technique, and 15 patients were treated with the mini external fixator (MEF) technique. Demographic data, previous treatment, pre- and postoperative carrying angle (CA), complications and additional procedures were recorded. Radiographic evaluation included assessment of the humerus-elbow-wrist angle (HEW), and the lateral prominence index (LPI). RESULTS: Patients treated with both KW and MEF achieved significant improvements in clinical alignment (mean pre-op CA -16 ± 6.1 degrees to mean post-op 8.9 ± 5.3 degrees, P < 0.001). There were no differences in final radiographic alignment or radiographic union time; however, time to achieve full elbow motion was faster in the MEF group (13.6 versus 34.3 weeks, P = 0.4547). Two patients (11.8%) in the KW group experienced complications, including one superficial infection and one failed correction that required unplanned revision surgery. Eleven patients in the MEF group underwent a planned second surgical procedure for hardware removal. CONCLUSIONS: Both fixation techniques are effective at correcting cubitus varus in the pediatric population. The MEF technique may have the advantage of shorter recovery of elbow range of motion but may require sedation for hardware removal. The KW technique may present a slightly higher complication rate.

Sustaining small-scale fisheries through a nation-wide Territorial Use Rights in Fisheries system.

Territorial Use rights in Fisheries (TURFs) are used around the world to manage small-scale fisheries and they've shown varying levels of success. Our understanding of what leads to different performance levels is limited due to several reasons. Firstly, these systems are often present in areas with low monitoring capacity where data is scarce. Secondly, past research has centered on the analysis of successful cases, with little attention paid to entire systems. Thirdly, research has been ahistorical, disconnected from the development process of TURF systems. Fourthly, TURFs are often viewed as homogenous ignoring the socio-ecological conditions under which they develop. To address these gaps, the study focuses on Mexico as a case study and context. The research first presents a historical overview of the development of TURF systems in Mexico, including the institutional and legal frameworks that have shaped their evolution. The paper then presents a TURF database that maps all TURF systems in Mexico, including their geographical locations and characteristics. In addition, the study presents case studies based on identified archetypes that showcase the diversity of TURF systems in Mexico, highlighting the different types of systems and the challenges they face. By presenting a comprehensive map of all TURF systems in Mexico, this research paper aims to make an important addition to the case studies in the global literature on TURF systems and provide a valuable resource for marine resource management policymakers, researchers, and practitioners.

Congenital bilateral clinical anophthalmia and brachygnathia superior in a fighting bull calf.

This study describes a case of a 20-day-old male fighting bull with bilateral clinical anophthalmia and brachygnathia superior whose dam was 12.5 years old and was mistakenly dewormed with ivermectin intramuscularly in the first third of gestation in a livestock farm. A macroscopic examination of the carcass was performed, with a special focus on the ocular components. Eyeball remains were found in both orbits and a histopathological examination was performed on them. Antibodies by serological study against bovine herpes virus-1, respiratory syncytial virus and bovine viral diarrhea virus for both the cow and the calf were not detected. The calf had small orbits and inside them a white and brown mass of soft consistency. Microscopically, abundant muscular and adipose tissue was observed, alongside nervous structures and vestiges of ocular structures with stratified epithelium and abundant connective tissue with glands. No evidence that this congenital bilateral anophthalmia had infectious or hereditary origin was found. By contrast, the malformation could be related to the treatment with ivermectin during the first month of gestation.

Blind deconvolution of second harmonic microscopy images of the living human eye.

Second harmonic generation (SHG) imaging microscopy of thick biological tissues is affected by the presence of aberrations and scattering within the sample. Moreover, additional problems, such as uncontrolled movements, appear when imaging in-vivo. Deconvolution methods can be used to overcome these limitations under some conditions. In particular, we present here a technique based on a marginal blind deconvolution approach for improving SHG images obtained in vivo in the human eye (cornea and sclera). Different image quality metrics are used to quantify the attained improvement. Collagen fibers in both cornea and sclera are better visualized and their spatial distributions accurately assessed. This might be a useful tool to better discriminate between healthy and pathological tissues, especially those where changes in collagen distribution occur.

Genetic analysis of resistance to bean leaf crumple virus identifies a candidate LRR-RLK gene.

Bean leaf crumple virus (BLCrV) is a novel begomovirus (family Geminiviridae, genus Begomovirus) infecting common bean (Phaseolus vulgaris L.), threatening bean production in Latin America. Genetic resistance is required to ensure yield stability and reduce the use of insecticides, yet the available resistance sources are limited. In this study, three common bean populations containing a total of 558 genotypes were evaluated in different yield and BLCrV resistance trials under natural infection in the field. A genome-wide association study identified the locus BLC7.1 on chromosome Pv07 at 3.31 Mbp, explaining 8 to 16% of the phenotypic variation for BLCrV resistance. In comparison, whole-genome regression models explained 51 to 78% of the variation and identified the same region on Pv07 to confer resistance. The most significantly associated markers were located within the gene model Phvul.007G040400, which encodes a leucine-rich repeat receptor-like kinase subfamily III member and is likely to be involved in the innate immune response against the virus. The allelic diversity within this gene revealed five different haplotype groups, one of which was significantly associated with BLCrV resistance. As the same genome region was previously reported to be associated with resistance against other geminiviruses affecting common bean, our study highlights the role of previous breeding efforts for virus resistance in the accumulation of positive alleles against newly emerging viruses. In addition, we provide novel diagnostic single-nucleotide polymorphism markers for marker-assisted selection to exploit BLC7.1 for breeding against geminivirus diseases in one of the most important food crops worldwide.

Superpixel-Based Optic Nerve Head Segmentation Method of Fundus Images for Glaucoma Assessment.

Glaucoma disease is the second leading cause of blindness in the world. This progressive ocular neuropathy is mainly caused by uncontrolled high intraocular pressure. Although there is still no cure, early detection and appropriate treatment can stop the disease progression to low vision and blindness. In the clinical practice, the gold standard used by ophthalmologists for glaucoma diagnosis is fundus retinal imaging, in particular optic nerve head (ONH) subjective/manual examination. In this work, we propose an unsupervised superpixel-based method for the optic nerve head (ONH) segmentation. An automatic algorithm based on linear iterative clustering is used to compute an ellipse fitting for the automatic detection of the ONH contour. The tool has been tested using a public retinal fundus images dataset with medical expert ground truths of the ONH contour and validated with a classified (control vs. glaucoma eyes) database. Results showed that the automatic segmentation method provides similar results in ellipse fitting of the ONH that those obtained from the ground truth experts within the statistical range of inter-observation variability. Our method is a user-friendly available program that provides fast and reliable results for clinicians working on glaucoma screening using retinal fundus images.

Dental Age Estimation Using Multiphoton Microscopy: A Potential Tool for Forensic Science.

Normal aging affects the different structures of teeth, in particular, the dentine. These changes are useful in forensic disciplines as a tool for age estimation. Although multiphoton (MP) microscopy has been used to explore dental pieces, a relationship between age and MP response of the human dentine has not been proposed yet. The relationship between MP signals and natural dentine aging is investigated herein. An index of age (INAG) combining two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) images has been used to quantify these changes. The results show that the INAG significantly decreases with age. Moreover, peritubular dentine size and collagen internal properties are also modified with age. This information confirms the usefulness of this technique in forensic age estimation after disasters (natural or manmade) with a lack of comprehensive fingerprint database. Courts and other government authorities might also benefit from this tool when the official age of individuals under special circumstances is required for legal or medical reasons.

Biomimetic Remineralization of an Extracellular Matrix Collagen Membrane for Bone Regeneration.

Natural extracellular matrix (ECM) collagen membranes are frequently used for bone regeneration procedures. Some disadvantages, such as rapid degradation and questionable mechanical properties, limit their clinical use. These membranes have a heterologous origin and may proceed from different tissues. Biomineralization is a process in which hydroxyapatite deposits mainly in collagen fibrils of the matrices. However, when this deposition occurs on the ECM, its mechanical properties are increased, facilitating bone regeneration. The objective of the present research is to ascertain if different membranes from distinct origins may undergo biomineralization. Nanomechanical properties, scanning electron (SEM) and multiphoton (MP) microscopy imaging were performed in three commercially available ECMs before and after immersion in simulated body fluid solution for 7 and 21 d. The matrices coming from porcine dermis increased their nanomechanical properties and they showed considerable mineralization after 21 d, as observed in structural changes detected through SEM and MP microscopy. It is hypothesized that the more abundant crosslinking and the presence of elastin fibers within this membrane explains the encountered favorable behavior.

Galvanic current activates the NLRP3 inflammasome to promote Type I collagen production in tendon.

The NLRP3 inflammasome coordinates inflammation in response to different pathogen- and damage-associated molecular patterns, being implicated in different infectious, chronic inflammatory, metabolic and degenerative diseases. In chronic tendinopathic lesions, different non-resolving mechanisms produce a degenerative condition that impairs tissue healing and which therefore complicates their clinical management. Percutaneous needle electrolysis consists of the application of a galvanic current and is an emerging treatment for tendinopathies. In the present study, we found that galvanic current activates the NLRP3 inflammasome and induces an inflammatory response that promotes a collagen-mediated regeneration of the tendon in mice. This study establishes the molecular mechanism of percutaneous electrolysis that can be used to treat chronic lesions and describes the beneficial effects of an induced inflammasome-related response.

Host-Pathogen Adhesion as the Basis of Innovative Diagnostics for Emerging Pathogens.

Infectious diseases are an existential health threat, potentiated by emerging and re-emerging viruses and increasing bacterial antibiotic resistance. Targeted treatment of infectious diseases requires precision diagnostics, especially in cases where broad-range therapeutics such as antibiotics fail. There is thus an increasing need for new approaches to develop sensitive and specific in vitro diagnostic (IVD) tests. Basic science and translational research are needed to identify key microbial molecules as diagnostic targets, to identify relevant host counterparts, and to use this knowledge in developing or improving IVD. In this regard, an overlooked feature is the capacity of pathogens to adhere specifically to host cells and tissues. The molecular entities relevant for pathogen-surface interaction are the so-called adhesins. Adhesins vary from protein compounds to (poly-)saccharides or lipid structures that interact with eukaryotic host cell matrix molecules and receptors. Such interactions co-define the specificity and sensitivity of a diagnostic test. Currently, adhesin-receptor binding is typically used in the pre-analytical phase of IVD tests, focusing on pathogen enrichment. Further exploration of adhesin-ligand interaction, supported by present high-throughput "omics" technologies, might stimulate a new generation of broadly applicable pathogen detection and characterization tools. This review describes recent results of novel structure-defining technologies allowing for detailed molecular analysis of adhesins, their receptors and complexes. Since the host ligands evolve slowly, the corresponding adhesin interaction is under selective pressure to maintain a constant receptor binding domain. IVD should exploit such conserved binding sites and, in particular, use the human ligand to enrich the pathogen. We provide an inventory of methods based on adhesion factors and pathogen attachment mechanisms, which can also be of relevance to currently emerging pathogens, including SARS-CoV-2, the causative agent of COVID-19.

Assessment of the corneal collagen organization after chemical burn using second harmonic generation microscopy.

The organization of the corneal stoma is modified due to different factors, including pathology, surgery or external damage. Here the changes in the organization of the corneal collagen fibers during natural healing after chemical burn are investigated using second harmonic generation (SHG) imaging. Moreover, the structure tensor (ST) was used as an objective tool for morphological analyses at different time points after burn (up to 6 months). Unlike control corneas that showed a regular distribution, the collagen pattern at 1 month of burn presented a non-organized arrangement. SHG signal levels noticeably decreased and individual fibers were hardly visible. Over time, the healing process led to a progressive re-organization of the fibers that could be quantified through the ST. At 6 months, the stroma distribution reached values similar to those of control eyes and a dominant direction of the fibers re-appeared. The present results show that SHG microscopy imaging combined with the ST method is able to objectively monitor the temporal regeneration of the corneal organization after chemical burn. Future implementations of this approach into clinically adapted devices would help to diagnose and quantify corneal changes, not only due to chemical damages, but also as a result of disease or surgical procedures.

The Trimeric Autotransporter Adhesin YadA of Yersinia enterocolitica Serotype O:9 Binds Glycan Moieties.

Yersinia adhesin A (YadA) is a key virulence factor of Yersinia enterocolitica and Yersinia pseudotuberculosis. YadA is a trimeric autotransporter adhesin, a class of adhesins that have been shown to enable many Gram-negative pathogens to adhere to/interact with the host extracellular matrix proteins such as collagen, vitronectin, and fibronectin. Here, we show for the first time that YadA of Yersinia enterocolitica serotype O:9 not only interacts with proteinaceous surface molecules but can also attach directly to glycan moieties. We show that YadA from Y. enterocolitica serotype O:9 does not interact with the vitronectin protein itself but exclusively with its N-linked glycans. We also show that YadA can target other glycan moieties as found in heparin, for example. So far, little is known about specific interactions between bacterial autotransporter adhesins and glycans. This could potentially lead to new antimicrobial treatment strategies, as well as diagnostic applications.

Arrangement of the photoreceptor mosaic in a diabetic rat model imaged with multiphoton microscopy.

Diabetic retinopathy (DR) is defined as a microvascular pathology. However, some data have suggested that the retinal photoreceptors (PRs) might be important in the pathogenesis of this ocular disease. In this study the organization of the PRs in control and diabetic-induced rats was compared using multiphoton microscopy. The PR mosaic was imaged at different locations in non-stained retinas. The density of PRs was directly quantified from cell counting. The spatially resolved density presents a double-slope pattern (from the central retina towards the periphery) in both healthy and pathological samples, although the values for the latter were significantly lower all across the retina. Moreover, Voronoi analysis was performed to explore changes in PR topography. In control specimens a hexagonally packed structure was dominant. However, despite the non-controlled effects of the disease in retinal structures, this PR regularity was fairly maintained in diabetic retinas.

Distal Femoral Physeal Bar Resection Combined With Guided Growth for the Treatment of Angular Limb Deformity Associated With Growth Arrest: A Preliminary Report.

PURPOSE: Distal femoral growth arrest can result in progressive deformities and functional disability. The treatment is challenging given the significant growth potential of the distal femoral physis. This study addresses the short-term outcomes after distal femoral physeal bar resection combined with guided growth for the treatment of angular limb deformity. METHODS: We conducted a retrospective analysis of patients treated with distal femoral physeal bar resection, fat graft interposition, and growth modulation with a tension-band plate. Data recorded included patient demographics, growth arrest cause, physeal bar size, time-to-surgery, details of the operative procedure, and complications. The mechanical axis zones, tibiofemoral angle, and the anatomic lateral distal femoral angle were assessed on 51-inch anteroposterior standing radiographs. RESULTS: Five patients (3 male individuals) with valgus (n=4) and varus deformities (n=1) due to physeal arrests of the distal femur were analyzed. The cause of the physeal arrest was trauma (n=3) and infection (n=2). The average age at the time of surgery was 6.6 years (range: 2 to 11 y). Average size of the physeal bar was 413.4 mm, which represented 16.8% of the total distal femoral physis (range: 12% to 26%). Four of the 5 patients had a total correction of the deformity in 14.3 months (range: 9 to 22 mo). One patient required correction by osteotomy and external fixation. Postoperatively, 1 patient presented no improvement, and 4 had restoration of the longitudinal bone growth and alignment. Two patients had rebound valgus: one is being observed and another has undergone a repeat guided growth procedure. CONCLUSIONS: Distal femoral physeal bar resection combined with tension-band hemiepiphysiodesis provides a viable option for the correction of angular deformities associated with physeal arrest. Longer follow-up is required to evaluate future growth of the distal femoral physis after this combined procedure. LEVEL OF EVIDENCE: Level IV-therapeutic study.

Objective analysis of collagen organization in thyroid nodule capsules using second harmonic generation microscopy images and the Hough transform.

Papillary carcinoma is the most prevalent type of thyroid cancer. Its diagnosis requires accurate and subjective analyses from expert pathologists. Here we propose a method based on the Hough transform (HT) to detect and objectively quantify local structural differences in collagen thyroid nodule capsules. Second harmonic generation (SHG) microscopy images were acquired on non-stained histological sections of capsule fragments surrounding the healthy thyroid gland and benign and tumoral/malignant nodules. The HT was applied to each SHG image to extract numerical information on the organization of the collagen architecture in the tissues under analysis. Results show that control thyroid capsule samples present a non-organized structure composed of wavy collagen distribution with local orientations. On the opposite, in capsules surrounding malignant nodules, a remodeling of the collagen network takes place and local undulations disappear, resulting in an aligned pattern with a global preferential orientation. The HT procedure was able to quantitatively differentiate thyroid capsules from capsules surrounding papillary thyroid carcinoma (PTC) nodules. Moreover, the algorithm also reveals that the collagen arrangement of the capsules surrounding benign nodules significantly differs from both the thyroid control and PTC nodule capsules. Combining SHG imaging with the HT results thus in an automatic and objective tool to discriminate between the pathological modifications that affect the capsules of thyroid nodules across the progressions of PTC, with potential to be used in clinical settings to complement current state-of-the-art diagnostic methods.

Spermcast mating with release of zygotes in the small dioecious bivalve Digitaria digitaria.

Digitaria digitaria, a small astartid usually less than 10 mm in length, has a non-brooding behaviour in spite of its limited space for gonad development. This species lives in highly unstable environments with strong currents, which represent a challenge for fertilization and larval settlement. The studied population of D. digitaria from the Strait of Gibraltar area was dioecious, with significant predominance of females and sexual dimorphism, where females are larger than males. The reproductive cycle is asynchronous throughout the year, without a resting period, but with successive partial spawning events. The presence of stored sperm in the suprabranchial chamber and inside the gonad of some females, together with the release of eggs along the dorsal axis of both gills, points to internal oocyte fertilization. Bacteriocytes were found in the female and male follicle walls, but no bacteria were observed inside any of the gametes. Digitaria digitaria could represent a "missing link" between spermcast mating bivalves with brooded offspring and bivalves with broadcast release of eggs and sperm. The small size, limiting the oocyte production, together with the unstable environment could represent evolutionary pressures towards sperm uptake in D. digitaria.

Fluorescent bicolour sensor for low-background neutrinoless double ß decay experiments.

Observation of the neutrinoless double ß decay is the only practical way to establish that neutrinos are their own antiparticles1. Because of the small masses of neutrinos, the lifetime of neutrinoless double ß decay is expected to be at least ten orders of magnitude greater than the typical lifetimes of natural radioactive chains, which can mimic the experimental signature of neutrinoless double ß decay2. The most robust identification of neutrinoless double ß decay requires the definition of a signature signal-such as the observation of the daughter atom in the decay-that cannot be generated by radioactive backgrounds, as well as excellent energy resolution. In particular, the neutrinoless double ß decay of 136Xe could be established by detecting the daughter atom, 136Ba2+, in its doubly ionized state3-8. Here we demonstrate an important step towards a 'barium-tagging' experiment, which identifies double ß decay through the detection of a single Ba2+ ion. We propose a fluorescent bicolour indicator as the core of a sensor that can detect single Ba2+ ions in a high-pressure xenon gas detector. In a sensor made of a monolayer of such indicators, the Ba2+ dication would be captured by one of the molecules and generate a Ba2+-coordinated species with distinct photophysical properties. The presence of such a single Ba2+-coordinated indicator would be revealed by its response to repeated interrogation with a laser system, enabling the development of a sensor able to detect single Ba2+ ions in high-pressure xenon gas detectors for barium-tagging experiments.

Quantitative Discrimination of Healthy and Diseased Corneas With Second Harmonic Generation Microscopy.

PURPOSE: To analyze the spatial organization of pathological corneas with second harmonic generation (SHG) imaging and to provide a proof of concept to objectively distinguish these from the healthy corneas. METHODS: A custom-built SHG microscope was used to image the anterior stroma of ex vivo corneas, both control and affected by some representative pathologies. The structure tensor (ST) was employed as a metric to explore and quantify the alterations in the spatial distribution of the collagen lamellae. RESULTS: The collagen arrangement differed between healthy and pathological samples. The former showed a regular distribution and a low structural dispersion (SD < 40°) within the stroma with a well-defined dominant orientation. This regular arrangement drastically turns into a disorganized pattern in pathological corneas (SD > 40°). CONCLUSIONS: The combination of SHG imaging and the ST allows obtaining quantitative information to differentiate the stromal collagen organization in healthy and diseased corneas. This approach represents a feasible and powerful technique with potential applications in clinical corneal diagnoses. TRANSLATIONAL RELEVANCE: The ST applied to SHG microscopy images of the corneal stroma provides an experimental objective score to differentiate control from pathological or damaged corneas. Future implementations of this technique in clinical environments might might be a promising tool in Ophthalmology, not only to diagnose and monitor corneal diseases, but also to follow-up surgical outcome.