Natural infection of Trypanosoma sp. in domestic sheep from Ecuador

ABSTRACT: Trypanosomosis in sheep is a hemoparasitic disease of worldwide interest due to its effects on the health of animals and the economic impact on producers, is caused by the protozoan Trypanosoma sp. In the present study, the occurrence of Trypanosoma sp. in sheep from the urban-rural area of the canton Salitre was determined, through an applied study with a qualitative descriptive prospective cross-sectional approach, carried out between October 1, 2019, and February 19, 2020. Blood samples were extracted from the jugular vein and analyzed by the blood smear method using the Giemsa and Diff-Quick staining techniques. Previously, in a first study realized between 2018 and 2019, 2 cases (2%) of Trypanosoma sp., 1 case of Babesia sp. (1%), and 4 cases of Anaplasma marginale (4%) were identified, but without the presence of symptoms of the disease. However, in this new research study, of 170 animals sampled from 5 herds and aged between 3 and 8 years, 34 (20%) were positive for Trypanosoma sp., 6 for Babesia sp. (3.52%) and 6 for A. marginale (3.52%) with coinfection between them. In this research, deteriorated clinical aspects and low hematological values were also determined in positive animals; besides of the total of positive cases, 25 presented symptoms of hemoparasitic disease, 3 sheep had abortions and 8 died. The results of this study showed that Trypanosoma sp. could already become an endemic parasitosis in sheep in the country, representing a serious problem of animal health.

Differential Expression of DSCAM Guides the Patterning of Retinal Axons Along Their Path and at Their Target in the Developing Xenopus Visual System

Background The Xenopus retinotectal circuit is organized topographically, where the dorsal-ventral axis of the retina maps respectively on to the ventral-dorsal axis of the tectum; axons from the nasal-temporal axis of the retina project respectively to the caudal-rostral axis of the tectum. Studies throughout the last two decades have shown that mechanisms involving molecular recognition of proper termination domains are at work guiding topographic organization. Such studies have shown that graded distribution of molecular cues is important for topographic mapping. However, the molecular cues organizing topography along the developing optic nerve, and as retinal axons cross the chiasm and navigate towards their target in the tectum, remain unknown. Down syndrome cell adhesion molecule (DSCAM) has been characterized as a key molecule in axon guidance, making it a strong candidate involved in the topographic organization of retinal fibers along the optic path.Methods Using a combination of whole-brain clearing and immunohistochemistry staining techniques we characterized DSCAM expression and the projection of ventral and dorsal retinal fibers starting from the eye, followed to the optic nerve into the chiasm, and into the terminal target in the optic tectum in Xenopus laevis tadpoles. We also assessed the effects of DSCAM on the establishment of retinotopic maps through spatially and temporally targeted DSCAM knockdown on retinal ganglion cells (RGCs) with axons innervating the optic tectum. Results Highest expression of DSCAM was localized to the ventral posterior region of the optic nerve and chiasm; this expression pattern coincides with ventral fibers derived from ventral RGCs. Downregulating DSCAM levels affected the segregation and proper sorting of medial axon fibers, derived from ventral RGCs, within the tectal neuropil, indicating that DSCAM plays a role in retinotopic organization. ConclusionThese findings together with the observation that DSCAM immunoreactivity accumulates on the primary dendrites of tectal neurons indicates that DSCAM exerts multiple roles in coordinating retinotopic order and connectivity in the developing vertebrate visual system.

Evaluation of renal biopsies in various kidney diseases with reference to staining

Renal diseases of different origin and nature may produce essentially similar disturbances of renal functions and may have clinical similarities and hence there was a need to classify renal diseases more scientifically. The basic approach was to correlate clinical signs and symptoms with histological changes in the tissue, using both simple and special staining techniques so as to reach to a definitive diagnosis.The present study was conducted on renal biopsy referred to pathology department. Criteria for successful biopsy were as follows-Adequate biopsy sample size, correct processing of specimen, informed interpretation and issue of an accurate report. A total of 29 renal biopsies were examined. In minimal change disease, only in 4 patients the glomerulus was sclerosed. Membranous glomerulonephritis comprised of the maximum number of cases (9/30). Total of 3 cases of renal biopsies revealed amyloidosis. Focal amyloid deposits with deposits either near the hilum or perivascular areas were found in 33.3% of cases, while extensive amyloid deposits were found in 33.3% of the cases.It is necessary to determine both the type of renal disease and the cause of the primary disorder in order to make the diagnosis and various staining techniques play a very helpful role. The likelihood that the biopsy specimen accurately reflects the type and severity of the underlying disease is directly related to both the diffuseness of the disease process and the amount of tissue examined.

Development of New Staining Procedures for Diagnosing Cryptosporidium spp. in Fecal Samples by Computerized Image Analysis

Interpretation errors may still represent a limiting factor for diagnosing Cryptosporidium spp. oocysts with the conventional staining techniques. Humans and machines can interact to solve this problem. We developed a new temporary staining protocol associated with a computer program for the diagnosis of Cryptosporidium spp. oocysts in fecal samples. We established 62 different temporary staining conditions by studying 20 experimental protocols. Cryptosporidium spp. oocysts were concentrated using the Three Fecal Test (TF-Test®) technique and confirmed by the Kinyoun method. Next, we built a bank with 299 images containing oocysts. We used segmentation in superpixels to cluster the patches in the images; then, we filtered the objects based on their typical size. Finally, we applied a convolutional neural network as a classifier. The trichrome modified by Melvin and Brooke, at a concentration use of 25%, was the most efficient dye for use in the computerized diagnosis. The algorithms of this new program showed a positive predictive value of 81.3 and 94.1% sensitivity for the detection of Cryptosporidium spp. oocysts. With the combination of the chosen staining protocol and the precision of the computational algorithm, we improved the Ova and Parasite exam (O&P) by contributing in advance toward the automated diagnosis.

Opening the Gap: Rare Lichens With Rare Cyanobionts – Unexpected Cyanobiont Diversity in Cyanobacterial Lichens of the Order Lichinales

The last decades of research led to a change in understanding of lichens that are now seen as self-sustaining micro-ecosystems, harboring diverse microbial organisms in tight but yet not fully understood relationships. Among the diverse interdependencies, the relationship between the myco- and photobiont is the most crucial, determining the shape, and ecophysiological properties of the symbiotic consortium. Roughly 10% of lichens associate with cyanobacteria as their primary photobiont, termed cyanolichens. Up to now, the diversity of cyanobionts of bipartite lichens resolved by modern phylogenetic approaches is restricted to the filamentous and heterocytous genera of the order Nostocales. Unicellular photobionts were placed in the orders Chroococcales, Pleurocapsales, and Chroococcidiopsidales. However, especially the phylogeny and taxonomy of the Chroococcidiopsidales genera remained rather unclear. Here we present new data on the identity and phylogeny of photobionts from cyanolichens of the genera Gonohymenia, Lichinella, Peccania, and Peltula from a broad geographical range. A polyphasic approach was used, combining morphological and cultivation-depending characteristics (microscopy, staining techniques, life cycle observation, baeocyte motility, and nitrogen fixation test) with phylogenetic analyses of the 16S rRNA and 16S–23S ITS gene region. We found an unexpectedly high cyanobiont diversity in the cyanobacterial lichens of the order Lichinales, including two new genera and seven new species, all of which were not previously perceived as lichen symbionts. As a result, we describe the novel unicellular Chroococcidiopsidales genera Pseudocyanosarcina gen. nov. with the species Pseudocyanosarcina phycocyania sp. nov. (from Peltula clavata, Australia) and Compactococcus gen. nov. with the species Compactococcus sarcinoides sp. nov. (from Gonohymenia sp., Australia) and the new Chroococcidiopsidales species Aliterella compacta sp. nov. (from Peltula clavata, Australia), Aliterella gigantea sp. nov. (from Peltula capensis; South Africa), Sinocapsa ellipsoidea sp. nov. (from Peccania cerebriformis, Austria), as well as the two new Nostocales species Komarekiella gloeocapsoidea sp. nov. (from Gonohymenia sp., Czechia) and Komarekiella globosa sp. nov. (from Lichinella cribellifera, Canary Islands, Spain). Our study highlights the role of cyanolichens acting as a key in untangling cyanobacterial taxonomy and diversity. With this study, we hope to stimulate further research on photobionts, especially of rare cyanolichens.

Paraganglioma of the Thyroid Gland: A Case Report and a Review of the Literature

Paragangliomas of the thyroid gland are rare and usually they originate from the inferior laryngeal paraganglia. In this case report, we describe the case of a 78-year-old woman who presented with an incidental finding of thyroid nodule dislocating the trachea. After a systemic and radiological evaluation, right lobo-isthmectomy was performed, and the definitive diagnosis of paraganglioma was reached. Diagnosis of these thyroidal lesions could be difficult due to their rarity, to their specific radiological aspects and the need of employing specific histological staining techniques. Once the definitive diagnosis is reached, patients should undergo a systemic and genetic evaluation. Surgery is the gold standard treatment; radiotherapy should be considered when aggressive behavior is suspected. Regular long-lasting follow-up should be proposed to these patients considering the unpredictable behavior of these lesions.

Supervised Learning With Perceptual Similarity for Multimodal Gene Expression Registration of a Mouse Brain Atlas

The acquisition of high quality maps of gene expression in the rodent brain is of fundamental importance to the neuroscience community. The generation of such datasets relies on registering individual gene expression images to a reference volume, a task encumbered by the diversity of staining techniques employed, and by deformations and artifacts in the soft tissue. Recently, deep learning models have garnered particular interest as a viable alternative to traditional intensity-based algorithms for image registration. In this work, we propose a supervised learning model for general multimodal 2D registration tasks, trained with a perceptual similarity loss on a dataset labeled by a human expert and augmented by synthetic local deformations. We demonstrate the results of our approach on the Allen Mouse Brain Atlas (AMBA), comprising whole brain Nissl and gene expression stains. We show that our framework and design of the loss function result in accurate and smooth predictions. Our model is able to generalize to unseen gene expressions and coronal sections, outperforming traditional intensity-based approaches in aligning complex brain structures.

Assessment of Tumor Cell Death After Percutaneous Ultrasound– Guided Radiofrequency Ablation of Breast Carcinoma: A Prospective Study

Background: Current trends in breast cancer treatment include the use of less aggressive surgeries to reduce morbidity, shorten hospital stays and improve cosmetic results. The aim of the study is to assess tumor cell viability after percutaneous ultrasound (US)-guided radiofrequency ablation (RFA) for small breast cancer by a combination of staining techniques. Methods: A prospective study was conducted at a single institution from 2013 to 2017. Twenty women with invasive ductal carcinoma of the breast measuring ≤ 20 mm were treated with US-guided RFA followed immediately by surgical resection. Tumor viability pre- and post-RFA was assessed with Hematoxylin and Eosin (H&E), Nicotinamide adenine dinucleotide (NADH), Succinate dehydrogenase (SDH), Cytochrome c oxidase (COX), Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and Cytokeratin 18 and 19 (CK18/CK19) staining techniques. Outcomes and correlation with the different techniques were evaluated with principal component analysis Cronbach’s alpha. Results: Oxidative enzymes in frozen sections showed loss of SDH and NADH in 13 of the 16 tumors (81%) and COX in 11 of the 13 tumors (84%). In paraffin-embedded tissues, CK18 was negative or markedly reduced in 98% and CK19 in 100% of the cases. Lack of evidence of cell death was seen in 3 cases where the maximum temperature achieved at the center of the tumor was ≤ 70ºC. The reliability and internal consistency between the different staining techniques was high (Cronbach’s alpha, 0.8), with concordance between the staining results of the oxidative enzymes and of CK18/CK19. Conclusion: Loss of tumor viability in small breast tumors after US-guided percutaneous RFA could be assessed in our series with different staining methods. CK18 and CK19 could be used in paraffin-embedded tissues as surrogate markers of tumor cell viability after immediate RFA.