MicroRNA 29 modulates ß-cell mitochondrial metabolism and insulin secretion via underlying miR-29-OXPHOS complex pathways.

AIM: MicroRNAs (miRNAs) regulate ß-cell function, and ß-cell mitochondria and insulin secretion are perturbed in diabetes. We aimed to identify key miRNAs regulating ß-cell mitochondrial metabolism and novel ß-cell miRNA-mitochondrial pathways. METHODS: TargetScan (http://www.targetscan.org/) was used to predict if 16 miRNAs implicated in ß-cell function target 27 cis-eGenes implicated in mitochondrial activity. The expression of candidate miRNAs and insulin secretion after 24 and 1 h pre-incubation in 2.8, 11.1- and 16.7-mM glucose was measured in clonal INS-1 832/13 ß-cells. MiR-29 silenced INS-1 832/13 cells were assessed for insulin secretion (glucose, pyruvate, and K+), target cis-eGene expression (Ndufv3 and Ndufa10 components of mitochondrial complex I (CI)), OXPHOS (CI-V) protein expression, and mitochondrial OXPHOS respiration/activity. The expression of differentially expressed miR-29 miRNAs was evaluated in Goto-Kakizaki (GK) rat, db/db mouse and type 2 diabetic (T2D) human islets, as well as NMRI mouse islets cultured under glucolipotoxic conditions. RESULTS: MiR-29, miR-15 and miR-124 were predicted to regulate ~20 cis-eGenes, while miR-29 alone was predicted to regulate ≥12 of these in rat and human species. MiR-29 expression and insulin secretion were reduced in INS-1 832/13 cells after 24 h in elevated glucose. MiR-29 knockdown increased all tested insulin secretory responses, Nudfv3, Ndufa10, complex I and II expression, and cellular mitochondrial OXPHOS. MiR-29 expression was reduced in db/db islets but increased in GK rat and T2D human islets. CONCLUSION: We conclude miR-29 is a key miRNA in regulating ß-cell mitochondrial metabolism and insulin secretion via underlying miR-29-OXPHOS complex pathways. Furthermore, we infer reduced miR-29 expression compensatorily enhances insulin secretion under glucotoxicity.

Primary third eyelid lymphoma with aggressive local behavior in a dog.

Lymphoma is the most common hematopoietic neoplasm in dogs. However, cases primarily involving the third eyelid are rarely reported in the species. The subtype most frequently described in this location is marginal zone lymphoma of the mucosa-associated lymphoid tissue, which has an indolent behavior. A 10-year-old intact female Poodle was presented with a 2-month history of a nodule in the left third eyelid for evaluation. This nodule was surgically excised; however, local recurrence was detected nineteen days postoperatively, with the formation of a mass that expanded and replaced the extraocular orbital tissues and infiltrated the eye. Euthanasia was elected due to the deteriorating clinical condition, and a necropsy was performed. Macroscopically, the mass infiltrated and expanded the eyelids with narrowing of the palpebral fissure, expanded into the orbital space, and infiltrated the cornea and bulbar conjunctiva. Histologically, the mass was composed of sheets of intermediate to large neoplastic lymphocytes, with frequent epitheliotropism and tingible bodies. Neoplastic lymphocytes were immunoreactive for PAX5, CD20, and CD79a and were not immunoreactive for CD3. The neoplasm was restricted to the left eyelid and orbit. A diagnosis of primary third eyelid lymphoma (extranodal) with immunophenotype B and locally aggressive behavior was made based on the macroscopic, histological, and immunohistochemical findings.

Angle Assessment for Upper Limb Rehabilitation: A Novel Light Detection and Ranging (LiDAR)-Based Approach.

The accurate measurement of joint angles during patient rehabilitation is crucial for informed decision making by physiotherapists. Presently, visual inspection stands as one of the prevalent methods for angle assessment. Although it could appear the most straightforward way to assess the angles, it presents a problem related to the high susceptibility to error in the angle estimation. In light of this, this study investigates the possibility of using a new approach to angle calculation: a hybrid approach leveraging both a camera and LiDAR technology, merging image data with point cloud information. This method employs AI-driven techniques to identify the individual and their joints, utilizing the cloud-point data for angle computation. The tests, considering different exercises with different perspectives and distances, showed a slight improvement compared to using YOLO v7 for angle calculation. However, the improvement comes with higher system costs when compared with other image-based approaches due to the necessity of equipment such as LiDAR and a loss of fluidity during the exercise performance. Therefore, the cost-benefit of the proposed approach could be questionable. Nonetheless, the results hint at a promising field for further exploration and the potential viability of using the proposed methodology.

Correlation between macroscopic and microscopic relaxation dynamics of water: Evidence for two liquid forms.

Water is vital for life, and without it, biomolecules and cells cannot maintain their structures and functions. The remarkable properties of water originate from its ability to form hydrogen-bonding networks and dynamics, which the connectivity constantly alters because of the orientation rotation of individual water molecules. Experimental investigation of the dynamics of water, however, has proven challenging due to the strong absorption of water at terahertz frequencies. In response, by employing a high-precision terahertz spectrometer, we have measured and characterized the terahertz dielectric response of water from supercooled liquid to near the boiling point to explore the motions. The response reveals dynamic relaxation processes corresponding to the collective orientation, single-molecule rotation, and structural rearrangements resulting from breaking and reforming hydrogen bonds in water. We have observed the direct relationship between the macroscopic and microscopic relaxation dynamics of water, and the results have provided evidence of two liquid forms in water with different transition temperatures and thermal activation energies. The results reported here thus provide an unprecedented opportunity to directly test microscopic computational models of water dynamics.

Pasteurella multocida polyserositis in growing-finishing pigs.

Pasteurella multocida is the main secondary bacterium isolated from cases of swine pneumonia. Although highly pathogenic strains of P. multocida have been associated with primary septic lesions and polyserositis in pigs, studies on this pathological presentation in naturally occurring cases are limited. The aim of this work was to characterize the clinical, pathological and molecular findings in cases of P. multocida polyserositis in growing-finishing pigs in a commercial farm in Brazil. The mean age of 17 investigated pigs was 120 days. Clinically, the disease was acute (11/17), with clinical signs of dyspnoea and apathy. Sudden death occurred in some animals (6/17). The main gross findings included fibrinous serositis affecting the abdominal and thoracic cavities (17/17), fibrinous pericarditis (15/17), marked cranioventral pulmonary consolidation (17/17) and splenic infarcts (3/17). P. multocida was isolated in all cases from systemic sites, including the pericardial sac and abdominal exudate. Molecular typing of genus and species was performed on four isolates, and all were characterized as P. multocida type A. Another five isolates were positive for the pathogenicity marker gene pfhA by polymerase chain reaction. This study reinforces the role of P. multocida as a cause of polyserositis in growing-finishing pigs.

Ophthalmic and immunopathological characterization of systemic infectious diseases in cats.

Ocular involvement in systemic diseases is frequent in cats; however, without concurrent clinical and ophthalmic examinations with gross and/or histologic analysis of the eye, these findings can be underdiagnosed. This article aims to provide gross, histologic, and immunohistochemical characteristics of ocular lesions from cats submitted to necropsy, focusing on those caused by systemic infectious agents. Cats that died due to a systemic infectious disease were selected based on necropsy diagnosis and presence of ocular lesions. Gross, histologic, and immunohistochemical findings were recorded. From April 2018 to September 2019, 849 eyes of 428 cats were evaluated. Histologic abnormalities were seen in 29% of cases, which were classified as inflammatory (41%), neoplastic (32%), degenerative (19%), and metabolic/vascular (8%). Macroscopic changes were present in one-third of eyes with histologic lesions. Of these, 40% were attributed to inflammatory or neoplastic diseases associated with infectious agents. The most important infectious agents causing ocular disease in this study were feline leukemia virus, feline infectious peritonitis virus, and Cryptococcus sp. The most common ocular abnormalities associated with infectious agents were uveitis (anterior, posterior, or panuveitis), optic neuritis, and meningitis of the optic nerve. Ocular lesions secondary to systemic infections in cats are frequent; however, these are not always diagnosed because gross lesions are less common than histologic lesions. Therefore, both gross and histologic evaluation of the eyes of cats is recommended, mainly for cases in which the clinical suspicion or necropsy diagnosis suggests that an infectious agent might be related to the cause of death.

A Machine Learning Approach to Robot Localization Using Fiducial Markers in RobotAtFactory 4.0 Competition.

Localization is a crucial skill in mobile robotics because the robot needs to make reasonable navigation decisions to complete its mission. Many approaches exist to implement localization, but artificial intelligence can be an interesting alternative to traditional localization techniques based on model calculations. This work proposes a machine learning approach to solve the localization problem in the RobotAtFactory 4.0 competition. The idea is to obtain the relative pose of an onboard camera with respect to fiducial markers (ArUcos) and then estimate the robot pose with machine learning. The approaches were validated in a simulation. Several algorithms were tested, and the best results were obtained by using Random Forest Regressor, with an error on the millimeter scale. The proposed solution presents results as high as the analytical approach for solving the localization problem in the RobotAtFactory 4.0 scenario, with the advantage of not requiring explicit knowledge of the exact positions of the fiducial markers, as in the analytical approach.

Motor and somatosensory degenerative myelopathy responsive to pantothenic acid in piglets.

This report describes 2 events of degenerative myelopathy in 4- to 27-day-old piglets, with mortality rates reaching 40%. Sows were fed rations containing low levels of pantothenic acid. Piglets presented with severe depression, weakness, ataxia, and paresis, which were more pronounced in the pelvic limbs. No significant gross lesions were observed. Histologically, there were degeneration and necrosis of neurons in the spinal cord, primarily in the thoracic nucleus in the thoracic and lumbar segments, and motor neurons in nucleus IX of the ventral horn in the cervical and lumbar intumescence. Minimal-to-moderate axonal and myelin degeneration was observed in the dorsal funiculus of the spinal cord and in the dorsal and ventral nerve roots. Immunohistochemistry demonstrated depletion of acetylcholine neurotransmitters in motor neurons and accumulation of neurofilaments in the perikaryon of neurons in the thoracic nucleus and motor neurons. Ultrastructurally, the thoracic nucleus neurons and motor neurons showed dissolution of Nissl granulation. The topographical distribution of the lesions indicates damage to the second-order neurons of the spinocerebellar tract, first-order axon cuneocerebellar tract, and dorsal column-medial lemniscus pathway as the cause of the conscious and unconscious proprioceptive deficit, and damage to the alpha motor neuron as the cause of the motor deficit. Clinical signs reversed and no new cases occurred after pantothenic acid levels were corrected in the ration, and piglets received parenteral administration of pantothenic acid. This study highlights the important and practical use of detailed neuropathological analysis to refine differential diagnosis.

Interfacial layers between ion and water detected by terahertz spectroscopy.

Dynamic fluctuations in the hydrogen-bond network of water occur from femto- to nanosecond timescales and provide insight into the structural/dynamical aspects of water at ion-water interfaces. Employing terahertz spectroscopy assisted with molecular dynamics simulations, we study aqueous chloride solutions of five monovalent cations, namely, Li, Na, K, Rb, and Cs. We show that ions modify the behavior of the surrounding water molecules and form interfacial layers of water around them with physical properties distinct from those of bulk water. Small cations with high charge densities influence the kinetics of water well beyond the first solvation shell. At terahertz frequencies, we observe an emergence of fast relaxation processes of water with their magnitude following the ionic order Cs > Rb > K > Na > Li, revealing an enhanced population density of weakly coordinated water at the ion-water interface. The results shed light on the structure breaking tendency of monovalent cations and provide insight into the properties of ionic solutions at the molecular level.

Eosinophilic Granulomatous Myocarditis in Two Pigs.

Eosinophilic myocarditis is a human condition that has been rarely documented in animals. We now report two unrelated porcine cases of idiopathic eosinophilic granulomatous myocarditis that resembled the human disease and which were associated with sudden death. The most relevant gross finding in both cases was marked cardiomegaly, accompanied by raised, multifocal to coalescent small white nodules (1-2 mm) and poorly demarcated multifocal pale areas in the epicardium. Histologically, there were multifocal to coalescent areas of cardiomyocyte loss with replacement by an intense inflammatory infiltrate of eosinophils and epithelioid macrophages, and proliferation of fibrous connective tissue. Immunohistochemistry for porcine circovirus type 2 (PCV2) and Toxoplasma gondii, in-situ hybridization and quantitative polymerase chain reaction tests for PCV2 and porcine circovirus type 3 and aerobic bacterial culture on myocardium samples were negative.

Probing Adaptation of Hydration and Protein Dynamics to Temperature.

Protein dynamics is strongly influenced by the surrounding environment and physiological conditions. Here we employ broadband megahertz-to-terahertz spectroscopy to explore the dynamics of water and myoglobin protein on an extended time scale from femto- to nanosecond. The dielectric spectra reveal several relaxations corresponding to the orientational polarization mechanism, including the dynamics of loosely bound, tightly bound, and bulk water, as well as collective vibrational modes of protein in an aqueous environment. The dynamics of loosely bound and bulk water follow non-Arrhenius behavior; however, the dynamics of water molecules in the tightly bound layer obeys the Arrhenius-type relation. Combining molecular simulations and effective-medium approximation, we have determined the number of water molecules in the tightly bound hydration layer and studied the dynamics of protein as a function of temperature. The results provide the important impact of water on the biochemical functions of proteins.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Duodenal leiomyosarcoma in a cat: Cytologic, pathologic, and immunohistochemical findings.

A 7-year-old mixed-breed neutered male cat was presented with a history of chronic vomiting with increased frequency in the last month, progressing to depression and profuse projectile vomiting. Abdominal ultrasonography revealed an area of marked segmental duodenal thickening associated with a nodule, and euthanasia was performed due to a poor prognosis. Grossly, at the postmortem examination, the wall of a duodenal segment was severely expanded by a nodule measuring 5.0 cm in diameter, resulting in marked luminal narrowing and obstruction. Smears were made by scraping the cut surface of the tumor during the postmortem examination. The smears were highly cellular and contained mesenchymal cells arranged individually or in large non-cohesive aggregates. Cells varied in morphology from spindle to round to stellate-shaped with marked anisocytosis and anisokaryosis. Numerous bi- and multinucleated neoplastic cells were observed. Histologic examination revealed a densely cellular neoplastic proliferation of mesenchymal cells that effaced the duodenal submucosa, muscularis externa, and serosa layers. Most cells were spindle-shaped; however, some tumor areas had numerous round cells and bi- and multinucleated neoplastic cells. Spindle cells showed variable cytoplasmic immunoreactivity for vimentin, smooth muscle actin, and desmin. Round, bi-, and multinucleated giant cells only had marked multifocal cytoplasmic immunoreactivity for vimentin. Neoplastic cells did not have immunoreactivity for cytokeratin, GFAP, S100, and CD117. A final diagnosis of duodenal leiomyosarcoma was made. This article reports the pathologic, cytologic, and immunohistochemical findings of this case and discusses the main differential diagnoses in cases of intestinal sarcomas in domestic animals.

An outbreak of visna-maedi in a flock of sheep in Southern Brazil.

Visna-maedi is a multisystemic and progressive inflammatory disease caused by a non-oncogenic retrovirus (Visna-maedi virus, VMV). An outbreak of visna-maedi occurred in Southern Brazil in sheep with clinical signs of blindness and stumbling gait. At post-mortem examination, all animals had similar lesions, including heavy non-collapsed lungs and multifocal yellow areas in the cerebral white matter, affecting mainly the periventricular region. These lesions corresponded histologically to lymphocytic interstitial pneumonia and histiocytic periventricular encephalitis surrounding areas of necrosis, in addition to significant demyelination in the brain. Serology was performed in all the sheep from the flock and 14% were seropositive for VMV. The presence of VMV was confirmed through PCR and partial sequencing of the 5'LTR. Sequencing demonstrated that the virus had 89.7 to 90.0% of nucleotide identity with VMV strains reported in the USA. This is the first description of clinical disease related to VMV in Brazil leading to economic losses. This study calls for the need to implement control measures to prevent the spread of small ruminant lentiviruses in Brazil.

Polypodium leucotomos targets multiple aspects of oral carcinogenesis and it is a potential antitumor phytotherapy against tongue cancer growth.

Introduction: Oral cancer refers to malignant tumors, of which 90% are squamous cell carcinomas (OSCCs). These malignancies exhibit rapid progression, poor prognosis, and often mutilating therapeutical approaches. The determination of a prophylactic and/or therapeutic antitumor role of the polyphenolic extract Polypodium leucotomos(PL) would be relevant in developing new tools for prevention and treatment. Methods: We aimed to determine the antitumor effect of PL by treating OSCC cell lines with PL metabolites and evaluating its action during OSCC progression in vivo. Results: PL treatment successfully impaired cell cycling and proliferation, migration, and invasion, enhanced apoptosis, and modulated macrophage polarization associated with the tumoral immune-inflammatory response of tongue cancer cell lines (TSCC). PL treatment significantly decreased the expression of MMP1 (p < 0.01) and MMP2 (p < 0.001), and increased the expression of TIMP1 (p < 0.001) and TIMP2 (p < 0.0001) in these cells. The mesenchymal-epithelial transition phenotype was promoted in cells treated with PL, through upregulation of E-CAD (p < 0.001) and reduction of N-CAD (p < 0.05). PL restrained OSCC progression in vivo by inhibiting tumor volume growth and decreasing the number of severe dysplasia lesions and squamous cell carcinomas. Ki-67 was significantly higher expressed in tongue tissues of animals not treated with PL(p < 0.05), and a notable reduction in Bcl2 (p < 0.05) and Pcna (p < 0.05) cell proliferation-associated genes was found in dysplastic lesions and TSCCs of PL-treated mice. Finally, N-cad(Cdh2), Vim, and Twist were significantly reduced in tongue tissues treated with PL. Conclusion: PL significantly decreased OSCC carcinogenic processes in vitro and inhibited tumor progression in vivo. PL also appears to contribute to the modulation of immune-inflammatory oral tumor-associated responses. Taken together, these results suggest that PL plays an important antitumor role in processes associated with oral carcinogenesis and may be a potential phytotherapeutic target for the prevention and/or adjuvant treatment of TSCCs.

Gold-black manufacture, microstructure, and optical characterization.

A previous contribution formulates a first-principle dipole antenna theory for predicting the polarization-sensitive directional spectral absorptance of gold-black in the near infrared. The current contribution chronicles a successful effort to validate that theory. After a brief review of gold-black history, we describe in some detail the design and construction of a vapor-deposition cell for laying down gold-black coatings on a mirrorlike gold substrate. The microstructure of 4- and 8-µm-thick coatings is revealed using scanning electron microscopy. An automated bench-level reflectometer has been used to measure the in-plane bidirectional reflectivity of the gold-black coatings in the visible (532 nm) and near-infrared (800 nm) for p and s polarization. Measurements are reported over incident zenith angles ranging between 10 and 50 deg. Results obtained using the apparatus are consistent with the dipole antenna theory in this range of incident zenith angles.