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Collective cell organization can be influenced by surface curvature, as studied in cylindrical, spherical, and sinusoidal surfaces; however, hyperbolic geometries have been overlooked due to their biofabrication complexity, despite their presence in biological structures, and their potential role in minimizing cell bending energy. Therefore, understanding cell behavior in complex-hyperbolic geometries is important to design in vitro biomimetic platforms and scaffolds. This work studies the role of curvature on collective cell spatiotemporal growth and adhesion, using the gyroid, a minimal periodic hyperbolic surface. The gyroid biofabrication has been limited by its interconnected curved structure, which is solved here by using its fundamental unit of symmetry, parametrized in terms of mean and Gaussian curvatures, achieving its biofabrication with>90% of resolution, allowing the exploration of its effect in guiding cell behavior, isolating the curvature variable from substrate stiffness and extracellular ligands. Three cell lines were studied: two epithelial-like (HEPG2 and HEK293), and one fibroblastic (NIH3T3), using superficial cell seeding and cell hydrogel encapsulation. It was observed by confocal microscopy that the gyroid was able to orient and guide the cells cytoskeleton in the direction to minimize their energy, showing less nuclei compression and eliciting more confluency than in semi-spheres and flat surfaces.

The upwelling hypothesis has been proposed to explain reduced or lack of population structure in seabird species specialized in food resources available at cold-water upwellings. However, population genetic structure may be challenging to detect in species with large population sizes, since variation in allele frequencies are more robust under genetic drift. High gene flow among populations, that can be constant or pulses of migration in a short period, may also decrease power of algorithms to detect genetic structure. Penguin species usually have large population sizes, high migratory ability but philopatric behavior, and recent investigations debate the existence of subtle population structure for some species not detected before. Previous study on Humboldt penguins found lack of population genetic structure for colonies of Punta San Juan and from South Chile. Here, we used mtDNA and nuclear markers (10 microsatellites and RAG1 intron) to evaluate population structure for 11 main breeding colonies of Humboldt penguins, covering the whole spatial distribution of this species. Although mtDNA failed to detect population structure, microsatellite loci and nuclear intron detected population structure along its latitudinal distribution. Microsatellite showed significant Rst values between most of pairwise locations (44 of 56 locations, Rst = 0.003 to 0.081) and 86% of individuals were assigned to their sampled colony, suggesting philopatry. STRUCTURE detected three main genetic clusters according to geographical locations: i) Peru; ii) North of Chile; and iii) Central-South of Chile. The Humboldt penguin shows signal population expansion after the Last Glacial Maximum (LGM), suggesting that the genetic structure of the species is a result of population dynamics and foraging colder water upwelling that favor gene flow and phylopatric rate. Our findings thus highlight that variable markers and wide sampling along the species distribution are crucial to better understand genetic population structure in animals with high dispersal ability.

AbstractsBackground/ObjectivesWe aim to describe the clinical and imaging characteristics of young children with dark without pressure- a well-defined area of dark retina in the mid periphery associated with attenuated hyperreflective outer segment bands on spectral domain OCT.Subjects/MethodsRetrospective medical chart review. We reviewed the charts and retinal images of children with dark without pressure (DWP) seen in the paediatric eye clinic that had undergone retinal imaging.ResultsWe identified 6 children (3 males, 3 females) ranging in age from 3 to 13 years with DWP lesions. These were an incidental finding in children referred for other disorders. The lesions spared the macula and were typically in the mid-peripheral retina. In those children who could cooperate with detailed imaging, the DWP lesions were hypoautofluorecent on fundus autofluorescence imaging and OCT imaging showed reduced reflectance of the outer segments and EZ line as has been described in older children and adults. In two of the six subjects a DWP lesion appeared in an area of previously normal fundus; in no subjects did the lesion resolve over the period of follow up.ConclusionsThe clinical and imaging characteristics of DWP lesions are similar in young children to those described in adults. The site of dysfunction appears to be at the level of the outer retina, but the precise cause is unknown. Recognition of this benign disorder will prevent patient anxiety and unnecessary further investigation.

Corynebacterium uterequi has been isolated from mares with infertility and early embryonic loss in mares from select herds of Europe and South Africa, but there is limited information regarding its occurrence and pathogenic role in other regions. This study described the laboratorial characteristic of uterine isolates of Corynebacterium sp. from mares with reproductive disorders in Rio Grande do Sul state (Brazil), and the associated cytological and histopathological findings. Considering the lack of data on the characteristics of this bacterial agent and its role in reproductive disorders in mares, microbiological culture and cytological and histopathological analyses were conducted in samples of thirty mares with reproductive disorders. Corynebacterium sp. was isolated from uterine samples of twelve mares, either as the sole bacterial isolate (n = 6) or in combination with other bacterial species (n = 6). Microbiological analysis of the isolates revealed morphological and biochemical profiles highly similar to C. uterequi. Molecular analysis of the 16S rRNA and rpoB gene regions from one isolate revealed high nucleotide identity - 99.38% and 100%, respectively–with C. uterequi sequences available in GenBank (accession number CP011546.1). Cytological evaluation revealed moderate to severe inflammation in eight of the twelve mares from which Corynebacterium sp. was isolated. Among these eight, inflammation was observed in two mares with Corynebacterium sp. only and in six mares where Corynebacterium sp. was isolated in combination with other bacteria. Histopathology of endometrial biopsies revealed glandular and periglandular fibrosis, periglandular nests, and glandular ectasia in two samples and lymphoplasmacytic endometritis in another. Additionally, two mares among the six that had Corynebacterium sp. as the sole isolate in uterine swabs presented multifocal periglandular fibrosis with nest formation and glandular ectasia. Hence, our study showed C. uterequi circulation in mares from Brazil and our findings contributed to the knowledge of selected aspects of this agent. Key words: horse, Gram-positive bacteria, reproductive losses, characterization, isolation. Corynebacterium uterequi tem sido isolado de éguas com infertilidade e perda embrionária precoce em rebanhos selecionados da Europa e África do Sul, porém há informações limitadas sobre sua ocorrência e papel patogênico em outras regiões. Este estudo teve como objetivo descrever as características laboratoriais de isolados uterinos de Corynebacterium sp. de éguas com distúrbios reprodutivos no Rio Grande do Sul (Brasil) e os achados citológicos e histopatológicos associados à infecção. Considerando a escassez de dados sobre as características deste agente bacteriano e seu papel nos distúrbios reprodutivos em éguas, foram realizadas análises microbiológicas, citológicas e histopatológicas em amostras de trinta éguas com histórico de distúrbios reprodutivos. Corynebacterium sp. foi isolado de amostras uterinas de doze éguas, estando associado (n = 6) ou não (n = 6) a outros agentes etiológicos. A análise microbiológica dos isolados revelou perfis morfológicos e bioquímicos altamente semelhantes aos de C. uterequi. A análise molecular das regiões do DNA 16S rRNA e rpoB de um dos isolados indicou alta identidade de nucleotídeos (99,38% e 100%, respectivamente) com C. uterequ depositado no GenBank (CP011546.1). Na citologia de oito éguas, foi observada inflamação de grau moderado a acentuado e entre esses oito animais, a inflamação foi observada em duas éguas com Corynebacterium sp. apenas e em seis éguas onde Corynebacterium sp. foi isolado em combinação com outras bactérias. A histopatologia das biópsias endometriais revelou fibrose glandular e periglandular, ninhos periglandulares e ectasia glandular em duas amostras, e endometrite linfoplasmocitária em outra. Duas éguas que apresentaram exclusivamente Corynebacterium sp. nos suabes uterinos exibiram fibrose periglandular multifocal com formação de ninhos e ectasia glandular. Assim, nosso estudo mostrou a circulação de C. uterequi em éguas do Brasil e nossos achados contribuem para o conhecimento de aspectos selecionados desse agente. Palavras-chave: cavalos, bactéria Gram-positiva, perdas reprodutivas, caracterização, isolamento bacteriano.

Background: Magnesium is a mineral that modulates several physiological processes. However, its relationship with intestinal microbiota has been scarcely studied. Therefore, this study aimed to assess the role of dietary magnesium content to modulate the intestinal microbiota of Wistar male rats. Methods: Rats were randomly assigned one of three diets: a control diet (C-Mg; 1000 mg/kg), a low magnesium content diet (L-Mg; 60 mg/kg), and a high magnesium content diet (H-Mg; 6000 mg/kg), for two weeks. After treatment, fecal samples were collected. Microbiota composition was assessed by sequencing the V3–V4 hypervariable region. Results: The C-Mg and L-Mg groups had more diversity than H-Mg group. CF231, SMB53, Dorea, Lactobacillus and Turibacter were enriched in the L-Mg group. In contrast, the phyla Proteobacteria, Parabacteroides, Butyricimonas, and Victivallis were overrepresented in the H-Mg group. PICRUSt analysis indicated that fecal microbiota of the L-Mg group were encoded with an increased abundance of metabolic pathways involving carbohydrate metabolism and butanoate metabolism. Conclusion: Dietary magnesium supplementation can result in intestinal dysbiosis development in a situation where there is no magnesium deficiency. Conversely, low dietary magnesium consumption is associated with microbiota with a higher capacity to harvest energy from the diet.

Infectious enteritis is highly prevalent among dogs worldwide and, in some cases, it can be fatal. This study describes the clinical and laboratorial findings of single and mixed infections associated with severe enteritis in 76 dogs from Southern Brazil. Intestinal segments and/or fecal samples were subjected to histopathology and molecular detection of DNA viruses, bacteria and protozoa. Severe intestinal lesions were observed in most cases. Single infections were detected in 52.6% of cases, double (36.8%) and triple (10.5%) infections were also identified. Carnivore protoparvovirus 2 (CPV-2) was the most frequent agent in single infections (36.8%). Coinfection by CPV-2 and Giardia spp. was the most common in dual infections (19.7%), followed by CPV-2 and Cryptosporidium spp. (10.5%). The most frequent triple infection was CPV-2, Giardia sp. and Cryptosporidium spp. (6.6%). Our results shown that single and mixed infections are associated with severe enteritis in dogs in southern Brazil, mainly involving CPV-2 and Giardia sp. Key words: gastroenteritis, dogs, bacteria, virus, protozoa, diagnosis. Enterites infecciosas são altamente prevalentes entre cães em todo o mundo e, em alguns casos, podem ser fatais. Este estudo descreve os achados clínicos e laboratoriais de infecções simples e mistas associadas a enterites graves em 76 cães do sul do Brasil. Segmentos intestinais e/ou fezes foram submetidos à análise patológica e detecção molecular de vírus de genoma DNA, bactérias e protozoários. Lesões intestinais severas foram observadas na maioria dos casos. Infecções únicas foram detectadas em 52,6% dos casos, e infecções duplas (36,8%) e triplas (10,5%) também foram identificadas. O Protoparvovírus carnívoro 2 (CPV-2) foi o agente mais frequente nas infecções simples (36,8% do total). Coinfecção por CPV-2 e Giardia sp. foi a mais comum em infecções duplas (19,7%), seguido por CPV-2 e Cryptosporidium spp. (10,5%). A tripla infecção mais frequente foi CPV-2, Giardia spp. e Cryptosporidium spp. (6,6%). Nossos resultados demostraram que infecções simples e mistas estão associadas a enterites graves em cães no sul do Brasil, envolvendo principalmente CPV-2 e Giardia sp. Palavras-chave: gastroenterite, cães, bactérias, vírus, protozoários, diagnóstico.

The necessity of developing more realistic artificial food requires the aggregation of different biomaterials in an ordered and controlled manner. One of the most advanced methods for this is food printers reproducing additive manufacturing processes. This study presents a fully automatized 3D food printer leveraging collaborative Cartesian and multi-ink robotic systems to create complex food structures, with materials with different rheological settings using a screw conveyor configuration with controlled motion velocity. The developed food printer followed a formal mechatronic design strategy with fully functional instrumentation and automation systems. An adaptive controller was developed and implemented to regulate the coordinated operation of booth robotic devices, which are enforced by the G-code corresponding to the target food structure, leading to the necessary resolution. This device was tested with different commercial food inks to develop structures with complex shapes. The workability of the developed printer was confirmed by examining the food samples obtained using multiple materials, including creating different three-dimensional structures of a single complex food ink and creating simple structures made of different food inks with diverse structures that could yield a synthetic tissue that reproduces synthetic meat.

Canine distemper virus (CDV) and canine circovirus (CanineCV) have been described worldwide in multi-systemic disease in dogs. Both agents may be occasionally associated with other viral pathogens, but reports of coinfection by CDV and CanineCV associated with disease are rare. In this article, we report a coinfection between CDV and CanineCV detected during an investigation of viral agents involved in multisystemic disease in dogs in Southern Brazil. Molecular testing by PCR in lungs and intestines of 77 dogs necropsied in pathology services (2015–2020) revealed several single and mixed viral infections, including a CDV/CanineCV coinfection. In the case reported here, gross and histological findings were compatible with CDV pathology (bronchointerstitial pneumonia and viral intracytoplasmatic inclusions in pneumocytes and transitional epithelial cells of urinary bladder). CanineCV DNA and CDV antigens were detected in lung and intestine fragments by PCR and immunohistochemistry, respectively. CanineCV PCR amplicons subjected to nucleotide sequencing showed > 98.6% nucleotide identity with CanineCV sequences from GenBank. Although the role of CanineCV in the pathogenesis of the reported case could not be determined, our results show that CanineCV may be associated with other viral infections in cases of multisystemic disease in dogs. These results reinforce the circulation of CanineCV in dogs in Brazil and highlight the importance of including this virus in the list of differential diagnoses of respiratory and gastroenteric infectious diseases in dogs.

Aldehyde dehydrogenase 1A1 (ALDH1A1) is a cancer stem cell (CSC) marker related to clinical outcomes in breast cancer (BC). The aim of this study was to analyze the relationship between ALDH1A1, programmed death ligand 1 (PD-L1) and tumor-infiltrating lymphocytes (TILs) in triple negative (TN) and human epidermal growth factor receptor 2-positive (HER2+) BC tumors, and its association with clinicopathological characteristics and outcomes. A retrospective, historical cohort study of patients diagnosed with early or locally advanced BC treated with neoadjuvant chemotherapy was conducted. ALDH1A1, PD-L1 expression and TILs were assessed using immunohistochemistry. A total of 75 patients were analyzed (42.7% TN, 57.3% HER2+ tumors). ALDH1A1+ was related to HTILs (p = 0.005) and PD-L1+ tumors (p = 0.004). ALDH1A1+ tumors presented higher CD3+ (p = 0.008), CD4+ (p = 0.005), CD8+ (p = 0.003) and CD20+ (p = 0.006) TILs. ALDH1A1+ (p = 0.018), PD-L1+ (p = 0.004) and HTILs (p < 0.001) were related to smaller tumors. ALDH1A1+ was related to pathologic complete response (pCR) (p = 0.048). At the end of the follow-up (54.4 [38.3–87.6] months), 47 patients (62.7%) remained disease-free, and 20 (26.7%) had died. HTILs were related to improved disease-free survival (p = 0.027). ALDH1A1+ was related to PD-L1+ and HITLs, that might be related to higher pCR rates with neoadjuvant therapy.