Poisson function and volume ratio of soft anisotropic materials under large deformations.

Accurate transverse deformation measurements are required for the estimation of the Poisson function and volume ratio. In this study, pure silicone and soft composite specimens were subjected to uniaxial tension, and the digital image correlation method was used to measure longitudinal and in- and out-of-plane transverse stretches. To minimize the effects of measurement errors on parameter estimation, the measured transverse stretches were defined in terms of the longitudinal stretch using a new formulation based on Poisson's ratios and two stretch-dependent parameters. From this formulation, Poisson functions and volume ratio for soft materials under large deformations were obtained. The results showed that pure silicone can be considered isotropic and nearly incompressible under large deformations, as expected. In contrast, Poisson's ratio of silicone reinforced with extensible fabric can exceed classical bounds, including negative value (auxetic behavior). The incompressibility assumption can be employed for describing the stress-stretch curve of pure silicone, while volume ratios are required for soft composites. Data of human skin, aortic wall, and annulus fibrosus from the literature were selected and analyzed. Except for the aortic wall, which can be considered nearly incompressible, the studied soft tissues must be regarded as compressible. All tissues presented anisotropic behavior.

Synergistic effects of glial cell line-derived neurotrophic factor and base-medium on in vitro culture of testicular tissue derived from prepubertal collared peccary.

We evaluated the influence of different media plus various concentrations of Glial cell line-derived neurotrophic factor (GDNF) during the in vitro culture (IVC) of testicular tissues from prepubertal collared peccary. Testes from 5 individuals were collected, fragmented and cultured for 28 days (34°C and 5% CO2). Culture media were Dulbecco's modified essential medium (DMEM) or stem cell serum free media (StemPro-34™ SFM), both supplemented with various concentrations of GDNF (0, 10, or 20 ng/mL). Fragments were cultured on the flat surface of 0.75% agarose gel and were evaluated every 7 days for fragment area, histomorphology, cellular viability, and proliferative activity. Data were expressed as mean ± standard error and analyzed by Kruskal-Wallis's and Tukey test. Fragments area decreased over the 28 days-culture, regardless of the treatment. For morphology, the StemPro-37 SFM medium plus 10 ng/mL GDNF provided higher scores at all time points in comparison to DMEM using any GDNF concentration (p < .05). After 28 days, similar cellular viability (~70%) was observed in all treatments (p > .05). For proliferating cell nuclear antigen assay, only DMEM plus 10 ng/mL GDNF improved (p < .05) cellular proliferation on Days 14 and 28. Looking at argyrophilic nucleolar organizing regions, after 28 days, there were no differences among treatments regarding cell proliferative capacity for both spermatogonia and Sertoli cells (p > .05). In summary, the DMEM and StemPro-34 SFM are adequate medium for IVC of prepubertal peccary testicular tissue. Supplementation with GDNF, especially at a 10 ng/mL concentration, appears to be essential for the maintenance of cell survival and proliferation.

Effectiveness of Percutaneous Needle Electrolysis to Reduce Pain in Tendinopathies: A Systematic Review With Meta-Analysis.

CONTEXT: Tendon injuries are common disorders in both workers and athletes, potentially impacting performance in both conditions. This is why the search for effective treatments is continuing. OBJECTIVE(S): The objective of this study was to analyze whether the ultrasound-guided percutaneous needle electrolysis technique may be considered a procedure to reduce pain caused by tendinosis. EVIDENCE ACQUISITION: The search strategy included the PubMed, SCOPUS, CINAHL, Physiotherapy Evidence Database, SciELO, and ScienceDirect up to the date of February 25, 2024. Randomized clinical trials that assessed pain caused by tendinosis using the Visual Analog Scale and Numeric Rating Scale were included. The studies were evaluated for quality using the Cochrane Risk of Bias 2, and the evidence strength was assessed by the GRADEpro GDT. EVIDENCE SYNTHESIS: Out of the 534 studies found, 8 were included in the review. A random-effects meta-analysis and standardized mean differences (SMD) were conducted. The ultrasound-guided percutaneous needle electrolysis proved to be effective in reducing pain caused by tendinosis in the overall outcome (SMD = -0.97; 95% CI, -1.26 to -0.68; I2 = 58%; low certainty of evidence) and in the short-term (SMD = -0.83, 95% CI, -1.29 to -0.38; I2 = 65%; low certainty of evidence), midterm (SMD = -1.28; 95% CI, -1.65 to -0.91; I2 = 0%; moderate certainty of evidence), and long-term (SMD = -0.94; 95% CI, -1.62 to -0.26; I2 = 71%; low certainty of evidence) subgroups. CONCLUSION(S): The application of the ultrasound-guided percutaneous needle electrolysis technique for reducing pain caused by tendinosis appears to be effective. However, due to the heterogeneity found (partially explained), more studies are needed to define the appropriate dosimetry, specific populations that may benefit more from the technique, and possible adverse events.

Bovine papillomavirus vertical transmission: BPV diversity and expression in maternal and fetal tissues.

Bovine Papillomaviruses (BPVs) constitute a diverse group within the Papillomaviridae family, playing a crucial role in bovine health and economic considerations. This study investigates the dynamics of vertical transmission of BPV in cattle, focusing on five cows and their reproductive tissues, as well as three gravid cows and their fetuses. DNA and RNA samples were extracted from the warts, fetal skin, placenta, uterus, ovary, and blood of cows, as well as the skin and blood of fetuses. Polymerase Chain Reaction (PCR) targeted BPV types 1-6 and 8-14, was assessed in both cows and fetuses. Additionally, Reverse Transcription PCR (RT-PCR) examined BPV-2 E5 oncogene expression in the skin and reproductive sites of mother cows and fetuses. Our findings unveil a rich diversity of BPV types, including BPV-2, 3, 9, 10, 12, and 14, present in both maternal and fetal tissues. Intriguingly, certain types, namely BPV-4, 6, 8, and 11, were exclusively identified in maternal tissues A higher diversity of BPVs was observed in cow warts, followed by cow blood, fetal blood, and fetal skin. Strikingly similar BPV types in gravid cow blood and fetuses suggest primary dissemination through the bloodstream and transmission via the placenta, though detected in lower numbers in cow uterus and ovary. Histopathological analysis revealed no abnormalities in the reproductive tissues despite the presence of BPV. However, in one bladder sample from a cow that did not consume bracken fern, urothelial neoplasia in situ was observed. The study extends beyond detection, exploring the expression of the BPV-2 E5 oncogene in fetal tissues, providing insights into potential cell implications. Comparative analyses with previous studies highlight the uniqueness of our investigation, encompassing a broader array of BPV types in the gravid cows and their fetuses. The findings not only establish a foundation for further investigations into the mechanisms of vertical transmission but also highlight the need for targeted interventions and surveillance strategies to mitigate potential health risks associated with specific BPV types.

Lower limb balance, ankle dorsiflexion, orofacial tissue pressure, and occlusal force of rugby players.

This cross-sectional study examined the lower limb balance, ankle dorsiflexion, orofacial tissue pressure, and occlusal strength of rugby players. Twenty-six participants were divided into groups: rugby players (n â€‹= â€‹13) and healthy sedentary adults (n â€‹= â€‹13). Participants underwent an analysis of lower limb balance using a composite score (Y-Balance Test). Ankle dorsiflexion was measured using the Lunge Test. The Iowa Oral Performance Instrument was employed to measure orofacial tissue pressure. Bite force was measured with a dynamometer, and T-Scan assessed occlusal contact distribution. Data were analyzed using the t-test (p â€‹< â€‹0.05) and ANCOVA with age and weight as covariates, where it is possible to verify that these factors did not influence the results obtained. Significant differences were observed in the balance of the right (p â€‹= â€‹0.07) and left (p â€‹= â€‹0.02) lower limbs, where rugby players had lower composite scores. There were significant differences in the right (p â€‹= â€‹0.005) and left (p â€‹= â€‹0.004) lunges, with rugby players showing lower values, as well as lower tongue pressure (p â€‹= â€‹0.01) and higher lip pressure (p â€‹= â€‹0.03), with significant differences to sedentary participants. There was no significant difference in molar bite force and distribution occlusal contacts between groups. Rugby seems to reduce lower limb displacement, cause ankle hypomobility, lead to changes in orofacial tissues, particularly the tongue and lips. This study is significant for identifying significant differences between rugby players and sedentary individuals, providing new insights into the impact of rugby on health and performance, which can benefit sports training and injury prevention.

Prognostic Factors Associated with Tumor Recurrence and Overall Survival in Soft Tissue Sarcomas of the Extremities in a Colombian Reference Cancer Center.

Introduction: Soft tissue sarcomas (STS) are low-incidence tumors whose clinical and histopathological factors are associated with adverse oncological outcomes. This study evaluated prognostic factors (PF) associated with tumor recurrence and overall survival (OS) in patients diagnosed with STS of the extremities, treated at the Instituto Nacional de Cancerología (INC), Bogotá, Colombia. Materials and Methods: An analytical observational study of a historical cohort was carried out, including patients diagnosed with STS and managed surgically in the Functional Unit for Breast and Soft Tissue Tumors of the INC from January 2008 to December 2018. Results: A total of 227 patients were included; 74.5% had tumors greater than 5 cm. Most patients (29.1%) were in stage IIIB at diagnosis. Age was associated with higher mortality (HR = 1.01; CI95%: 1-1.02; p = 0.048). Tumor persistence at admission to the INC (HR = 2.34; CI95%: 1.25-4.35; p = 0.007) and histologic grade III (HR = 5.36; CI95%: 2.29-12.56; p = <0.001) showed statistical significance in the multivariate analysis for recurrence of any type, as did the PFs associated with a higher risk of local recurrence (HR = 2.85; CI95%: 1.23-6.57; p = 0.014 and HR = 6.09; CI95%: 2.03-18.2; p = 0.001), respectively. Tumor size (HR = 1.03; CI95%: 1-1.06; p = 0.015) and histologic grade III (HR = 4.53; CI95%: 1.42-14.49; p = 0.011) were associated with a higher risk of distant recurrence. Conclusions: This cohort showed that in addition to histologic grade and tumor size, tumor persistence at the time of admission has an impact on disease recurrence, so STS should be managed by a multidisciplinary team with experience in this pathology in high-volume reference centers.

Giant cell tumor of paralaryngeal soft tissues, extending to the laryngeal cartilage.

Paralaryngeal mass with secondary extension to the thyroid cartilage involving confluent cysts, subjected to conservative surgical treatment with a diagnosis of giant cell tumor of soft tissues, a neoplasm morphologically similar but genetically unrelated to osseous giant cell tumors.

Influence of keratinized mucosa on peri-implant diseases: a retrospective cohort study in humans.

OBJECTIVE: The present study aimed to assess the relationship between keratinized mucosa width and peri-implant diseases, namely peri-implant mucositis and peri-implantitis. MATERIALS AND METHODS: Ninety-one dental implants in function for ≥ 6 months from 40 partially or completely edentulous non-smoker subjects (24 females and 16 males) were evaluated clinically and radiographically. The width of keratinized mucosa, probing depth, plaque index, bleeding on probing, and marginal bone levels were assessed. Keratinized mucosa width was categorized as ≥ 2 mm or < 2 mm. RESULTS: There was no statistically significant association between keratinized buccal mucosa width and peri-implant mucositis or peri-implantitis (p ≥ 0.37). In the regression analysis, peri-implantitis was associated with longer implant function time (RR: 2.55, 95% CI: 1.25-11.81, p = 0.02) and implants in the maxilla (RR: 3.15, 95% CI: 1.61-14.93, p = 0.003). Mucositis was not associated with any of the factors analyzed. CONCLUSION: In conclusion, in the present sample, keratinized buccal mucosa width was not associated with peri-implant diseases, suggesting that a band of keratinized mucosa may not be necessary to maintain peri-implant health. Prospective studies are required to better understand its role in the maintenance of peri-implant health.

Mesenchymal Stromal Cells from Perinatal Tissues as an Alternative for Ex Vivo Expansion of Hematopoietic Progenitor and Stem Cells from Umbilical Cord Blood.

Umbilical cord blood (UCB) serves as a source of hematopoietic stem and progenitor cells (HSPCs) utilized in the regeneration of hematopoietic and immune systems, forming a crucial part of the treatment for various benign and malignant hematological diseases. UCB has been utilized as an alternative HSPC source to bone marrow (BM). Although the use of UCB has extended transplantation access to many individuals, it still encounters significant challenges in selecting a histocompatible UCB unit with an adequate cell dose for a substantial proportion of adults with malignant hematological diseases. Consequently, recent research has focused on developing ex vivo expansion strategies for UCB HSPCs. Our results demonstrate that co-cultures with the investigated mesenchymal stromal cells (MSCs) enable a 10- to 15-fold increase in the cellular dose of UCB HSPCs while partially regulating the proliferation capacity when compared to HSPCs expanded with early acting cytokines. Furthermore, the secretory profile of UCB-derived MSCs closely resembles that of BM-derived MSCs. Moreover, both co-cultures exhibit alterations in cytokine secretion, which could potentially impact HSPC proliferation during the expansion process. This study underscores the fact that UCB-derived MSCs possess a remarkably similar supportive capacity to BM-derived MSCs, implying their potential use as feeder layers in the ex vivo expansion process of HSPCs.

Endophytic bacterial community structure and diversity of the medicinal plant Mirabilis himalaica from different locations.

Endophytic bacteria play important roles in medicinal plant growth, abiotic stress, and metabolism. Mirabilis himalaica (Edgew.) Heimerl is known for its medicinal value as Tibetan traditional plant; however, little is known about the endophytic bacteria associated with this plant in different geographic conditions and vegetal tissues. To compare the endophytic bacterial community associated with this plant in different geographic conditions and vegetal tissues, we collected the leaves, stems, and roots of M. himalaica from five locations, Nongmu college (NM), Gongbujiangda (GB), Zhanang County (ZL), Lang County (LX), and Sangri County (SR), and sequenced the 16S rRNA V5-V7 region with the Illumina sequencing method. A total of 522,450 high-quality sequences and 4970 operational taxonomic units (OTUs) were obtained. The different tissues from different locations harbored unique bacterial assemblages. Proteobacteria and Actinobacteria were the dominant phyla in all the samples, while the dominant genera changed based on the different tissues. The endophytic bacterial structures in the leaf and stem tissues were different compared to root tissues. Redundancy analysis (RDA) showed that the endophytic bacterial community was significantly correlated with pH, available phosphorus (AP), total phosphorus (TP), total nitrogen (TN), and soil organic matter (SOM). These findings suggested that the geographic conditions, climate type, ecosystem type, and tissues determined the endophytic bacterial composition and relative abundances. This conclusion could facilitate an understanding of the relationship and ecological function of the endophytic bacteria associated with M. himalaica and provide valuable information for artificial planting of M. himalaica and identifying and applying functional endophytic bacteria.

Novel ultra-stretchable and self-healing crosslinked poly (ethylene oxide)-cationic guar gum hydrogel.

Hydrogels are three-dimensional structures with specific features that render them useful for biomedical applications, such as tissue engineering scaffolds, drug delivery systems, and wound dressings. In recent years, there has been a significant increase in the search for improved mechanical properties of hydrogels derived from natural products to extend their applications in various fields, and there are different methods to obtain strengthened hydrogels. Cationic guar gum has physicochemical properties that allow it to interact with other polymers and generate hydrogels. This study aimed to develop an ultra-stretchable and self-healing hydrogel, evaluating the influence of adding PolyOX [poly(ethylene oxide)] on the mechanical properties and the interaction with cationic guar gum for potential tissue engineering applications. We found that variations in PolyOX concentrations and pH changes influenced the mechanical properties of cationic guar gum hydrogels. After optimization experiments, we obtained a novel hydrogel, which was semi-crystalline, highly stretchable, and with an extensibility area of approximately 400 cm2, representing a 33-fold increase compared to the hydrogel before being extended. Moreover, the hydrogel presented a recovery of 96.8% after the self-healing process and a viscosity of 153,347 ± 4,662 cP. Therefore, this novel hydrogel exhibited optimal mechanical and chemical properties and could be suitable for a broad range of applications in different fields, such as tissue engineering, drug delivery, or food storage.

Analysis of the geometrical influence of ring-opening samples on arterial circumferential residual stress reconstruction.

This work consists of analyzing the impact of geometrical features (thickness and curvature) on the estimation of circumferential residual stresses in arteries. For this purpose, a specific sample of lamb abdominal artery is chosen for analysis and, through computational tools based on Python libraries, the stress-free geometry is captured after the ring opening test. Numerical simulations are then used to reconstruct the sample in order to estimate the circumferential residual stresses. Then, four stress-free geometry models are analyzed: an ideal geometry, i.e., constant curvature and thickness; a constant curvature and variable thickness geometry; a variable curvature and constant thickness geometry; and a variable curvature and thickness geometry. The numerical results show that models perform well from a geometric point of view, where the most different feature was the closed outer perimeter that differs about 14% from the closed real sample. As far as residual stress is concerned, differences up to 198% were found in more realistic models taking a constant curvature and thickness model as reference. Thus, the analysis of a realistic geometry with highly variable curvature and thickness can introduce, compared to an idealized geometry, significant differences in the estimation of residual stresses. This could indicate that the characterization of arterial residual stresses is not sufficient when considering only the opening angle and, therefore, it is also necessary to incorporate more geometrical variables.

Commitment of human mesenchymal stromal cells to skeletal lineages is independent of their morphogenetic capacity.

BACKGROUND: Mesenchymal stromal cells (MSCs) are multipotent cell populations obtained from fetal and adult tissues. They share some characteristics with limb bud mesodermal cells such as differentiation potential into osteogenic, chondrogenic, and tenogenic lineages and an embryonic mesodermal origin. Although MSCs differentiate into skeletal-related lineages in vitro, they have not been shown to self-organize into complex skeletal structures or connective tissues, as in the limb. In this work, we demonstrate that the expression of molecular markers to commit MSCs to skeletal lineages is not sufficient to generate skeletal elements in vivo. AIM: To evaluate the potential of MSCs to differentiate into skeletal lineages and generate complex skeletal structures using the recombinant limb (RL) system. METHODS: We used the experimental system of RLs from dissociated-reaggregated human placenta (PL) and umbilical cord blood (UCB) MSCs. After being harvested and reaggregated in a pellet, cultured cells were introduced into an ectodermal cover obtained from an early chicken limb bud. Next, this filled ectoderm was grafted into the back of a donor chick embryo. Under these conditions, the cells received and responded to the ectoderm's embryonic signals in a spatiotemporal manner to differentiate and pattern into skeletal elements. Their response to differentiation and morphogenetic signals was evaluated by quantitative polymerase chain reaction, histology, immunofluorescence, scanning electron microscopy, and in situ hybridization. RESULTS: We found that human PL-MSCs and UCB-MSCs constituting the RLs expressed chondrogenic, osteogenic, and tenogenic molecular markers while differentially committing into limb lineages but could not generate complex structures in vivo. MSCs-RL from PL or UCB were committed early to chondrogenic lineage. Nevertheless, the UCB-RL osteogenic commitment was favored, although preferentially to a tenogenic cell fate. These findings suggest that the commitment of MSCs to differentiate into skeletal lineages differs according to the source and is independent of their capacity to generate skeletal elements or connective tissue in vivo. Our results suggest that the failure to form skeletal structures may be due to the intrinsic characteristics of MSCs. Thus, it is necessary to thoroughly evaluate the biological aspects of MSCs and how they respond to morphogenetic signals in an in vivo context. CONCLUSION: PL-MSCs and UCB-MSCs express molecular markers of differentiation into skeletal lineages, but they are not sufficient to generate complex skeletal structures in vivo.

Cephalometric Characteristics in Chilean Latino Population with Normal Occlusion and Harmonic Profiles in Permanent Dentition. A Retrospective Study / Características Cefalométricas en Población Latina Chilena con Oclusión Normal y Perfiles Armónicos en Dentición Permanente. Un Estudio Retrospectivo

SUMMARY: Malocclusion is usually treated based on clinical decisions complemented with a cephalometric analysis, allowing the comparison of an individual with standard reference norms. Cephalometric standards have mostly been obtained from Caucasian population, but may not be appropriate for other ethnic groups, becoming a clinically relevant problem in multicultural and multiracial societies. The present study aimed to establish cephalometric norms for Chilean-Latino population, using a representative sample of class I individuals in permanent dentition. A sample of 72 cephalometric x-rays of class I growing individuals (47 women and 25 men) between 10 and 20 years of age with class I occlusion and harmonic profile was obtained from the records of the Universidad de los Andes taken between 2012 and 2019, including 1164 individuals. The radiographs were classified according to their cervical vertebral maturation status, and cephalometrically analyzed, obtaining vertical and sagittal parameters in soft and hard tissues, which were compared with Caucasian cephalometric norms. The statistical analysis was performed using descriptive and inferential statistics (T-test, ANOVA and Bonferroni tests). Cephalometric norms were obtained for hard and soft tissues. Upon comparison with Caucasian norms, the subjects included in the sample present a tendency towards a convex profile, significant incisal proclination, dental protrusion, labial biprotrusion and an acute nasolabial angle. There are cephalometric differences between the Caucasian cephalometric norms and those observed Chilean Latino population, displaying differences at a hard and soft tissue level that should be taken into account for clinical decision making in Orthodontics.

La maloclusión generalmente se trata con base en decisiones clínicas complementadas con un análisis cefalométrico, lo que permite la comparación de un individuo con normas de referencia estándar. Los estándares cefalométricos se han obtenido en su mayoría de población caucásica, pero pueden no ser apropiados para otros grupos étnicos, convirtiéndose en un problema clínicamente relevante en sociedades multiculturales y multirraciales. El presente estudio tuvo como objetivo establecer normas cefalométricas para población chileno-latina, utilizando una muestra representativa de individuos clase I en dentición permanente. Se obtuvo una muestra de 72 radiografías cefalométricas de individuos en crecimiento clase I (47 mujeres y 25 hombres) entre 10 y 20 años de edad con oclusión clase I y perfil armónico de los registros de la Universidad de los Andes tomados entre 2012 y 2019, incluidas 1164 personas. Las radiografías se clasificaron según su estado de maduración vertebral cervical, y se analizaron cefalométricamente, obteniendo parámetros verticales y sagitales en tejidos blandos y duros, que se compararon con normas cefalométricas caucásicas. El análisis estadístico se realizó mediante estadística descriptiva e inferencial (T-test, ANOVA y pruebas de Bonferroni). Se obtuvieron normas cefalométricas para tejidos duros y blandos. En comparación con las normas caucásicas, los sujetos incluidos en la muestra presentan una tendencia hacia un perfil convexo, proinclinación incisal significativa, protrusión dental, biprotrusión labial y un ángulo nasolabial agudo. Existen diferencias entre las normas cefalométricas caucásicas y las observadas en población latina chilena, mostrando diferencias a nivel de tejidos duros y blandos que se deben considerar para la toma de decisiones clínicas en Ortodoncia.

Lesões periapicais de origem não-endodôntica: uma revisão integrativa

O objetivo deste trabalho foi elencar quais são as principais lesões periapicais que não têm origem a partir da necrose pulpar. Para tanto, realizou-se uma revisão integrativa da literatura, com busca nas bases de dados Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS), Medical Literature Analysis and Retrievel System Online (MEDLINE) e Bibliografia Brasileira de Odontologia (BBO). Os descritores utilizados foram "doenças periapicais" e "diagnóstico diferencial", combinados com o operador booleano "AND". Foram encontrados 42 artigos que responderam aos critérios de inclusão e exclusão. A análise dos estudos incluídos mostrou que as principais lesões periapicais de origem não endodôntica são o queratocisto odontogênico e o ameloblastoma. Embora a prevalência de lesões periapicais não endodônticas seja baixa, é importante estar atento aos possíveis diagnósticos diferenciais, visto que as lesões que mais apareceram na literatura se tratam de patologias localmente agressivas e potencialmente mutiladoras. Ademais, a conduta do profissional que se depara com qualquer tipo de lesão periapical que apresente comportamento fora dos padrões de normalidade deve ser a realização de biópsia e análise histopatológica.

The aim of this study was to list the main periapical lesions that do not originate from pulpal necrosis. An integrative literature review was carried out, with a search in the Latin American and Caribbean Literature in Health Sciences (LILACS), Medical Literature Analysis and Retrievel System Online (MEDLINE) and Bibliografia Brasileira de Odontologia (BBO) databases. The keywords used were "periapical diseases" and "differential diagnosis", combined with the boolean operator "AND". 42 articles were found that met the inclusion and exclusion criteria. The analysis of the included studies showed that the main periapical lesions of non-endodontic origin are the odontogenic keratocyst and the ameloblastoma. Although the prevalence of non-endodontic periapical lesions is low, it is important to be aware of possible differential diagnoses, since the lesions that most appeared in the literature are locally aggressive and potentially mutilating pathologies. In addition, the conduct of the professional who is faced with any type of periapical lesion that presents behavior outside the normal range should be to perform a biopsy and histopathological analysis.

Application of Real-Time PCR Assays for the Diagnosis of Histoplasmosis in Human FFPE Tissues Using Three Molecular Targets.

Histoplasmosis is a fungal infection caused by the thermally dimorphic fungus Histoplasma capsulatum. This infection causes significant morbidity and mortality in people living with HIV/AIDS, especially in countries with limited resources. Currently used diagnostic tests rely on culture and serology but with some limitations. No molecular assays are commercially available and the results from different reports have been variable. We aimed to evaluate quantitative real-time PCR (qPCR) targeting three protein-coding genes of Histoplasma capsulatum (100-kDa, H and M antigens) for detection of this fungus in formalin-fixed paraffin-embedded (FFPE) samples from patients with proven histoplasmosis. The sensitivity of 100-kDa, H and M qPCR assays were 93.9%, 91% and 57%, respectively. The specificity of 100-kDa qPCR was 93% when compared against samples from patients with other mycoses and other infections, and 100% when samples from patients with non-infectious diseases were used as controls. Our findings demonstrate that real-time PCR assays targeting 100-kDa and H antigen showed the most reliable results and can be successfully used for diagnosing this mycosis when testing FFPE samples.

Nfinder: automatic inference of cell neighborhood in 2D and 3D using nuclear markers.

BACKGROUND: In tissues and organisms, the coordination of neighboring cells is essential to maintain their properties and functions. Therefore, knowing which cells are adjacent is crucial to understand biological processes that involve physical interactions among them, e.g. cell migration and proliferation. In addition, some signaling pathways, such as Notch or extrinsic apoptosis, are highly dependent on cell-cell communication. While this is straightforward to obtain from membrane images, nuclei labelling is much more ubiquitous for technical reasons. However, there are no automatic and robust methods to find neighboring cells based only on nuclear markers. RESULTS: In this work, we describe Nfinder, a method to assess the cell's local neighborhood from images with nuclei labeling. To achieve this goal, we approximate the cell-cell interaction graph by the Delaunay triangulation of nuclei centroids. Then, links are filtered by automatic thresholding in cell-cell distance (pairwise interaction) and the maximum angle that a pair of cells subtends with shared neighbors (non-pairwise interaction). We systematically characterized the detection performance by applying Nfinder to publicly available datasets from Drosophila melanogaster, Tribolium castaneum, Arabidopsis thaliana and C. elegans. In each case, the result of the algorithm was compared to a cell neighbor graph generated by manually annotating the original dataset. On average, our method detected 95% of true neighbors, with only 6% of false discoveries. Remarkably, our findings indicate that taking into account non-pairwise interactions might increase the Positive Predictive Value up to + 11.5%. CONCLUSION: Nfinder is the first robust and automatic method for estimating neighboring cells in 2D and 3D based only on nuclear markers and without any free parameters. Using this tool, we found that taking non-pairwise interactions into account improves the detection performance significantly. We believe that using our method might improve the effectiveness of other workflows to study cell-cell interactions from microscopy images. Finally, we also provide a reference implementation in Python and an easy-to-use napari plugin.

Genotyping of dengue virus from infected tissue samples embedded in paraffin.

Dengue has become one of the vector-borne diseases that affect humans worldwide. In Latin American countries, Colombia is historically one of the most affected by epidemics of this flavivirus. The underreporting of signs and symptoms of probable cases of dengue, the lack of characterization of the serotypes of the infection, and the few detailed studies of postmortem necropsies of patients are among other conditions that have delayed progress in the knowledge of the pathogenesis of the disease. This study presents the results of fragment sequencing assays on paraffin-embedded tissue samples from fatal DENV cases during the 2010 epidemic in Colombia. We found that the predominant serotype was DENV-2, with the Asian/American genotype of lineages 1 and 2. This work is one of the few reports of the circulating genotypes of dengue during the 2010 epidemic in Colombia, one of the most lethal dates in the country's history.

Proposing Clinicopathological Staging and Mitotic Count as Prognostic Factors for Canine Soft Tissue Sarcomas.

Soft tissue sarcomas (STSs) are a heterogeneous group of malignant mesenchymal tumors with similar histological features and biological behaviors. They are characterized by a low to moderate local recurrence rate and low metastasis, affecting approximately 20% of patients. Although this tumor set is vital in veterinary medicine, no previous unified staging system or mitotic count has been associated with patient prognosis. Therefore, this study proposed a new clinicopathological staging method and evaluated a cut-off value for mitosis related to the survival of dogs affected by STS. This study included 105 dogs affected by STS, treated only with surgery, and a complete follow-up evaluation. The new clinicopathological staging system evaluated tumor size (T), nodal involvement (N), distant metastasis (M), and histological grading criteria (G) to categorize the tumor stage into four groups (stages I, II, III, and IV). The proposed tumor staging system was able to differentiate patients' prognoses, with dogs with stage IV disease experiencing the lowest survival time and dogs with stage I disease having the highest survival time (p < 0.001). Moreover, we assessed the median mitosis (based on mitotic count) and its association with overall survival. Our study's median mitosis was 5, and patients with ≤5 mitoses had a higher survival time (p = 0.006). Overall, the proposed staging system and mitotic count seemed promising in the prediction of patient prognosis.

Normal tissue homeostasis and impairment of selective inflammatory responses in dendritic cells deficient for ATF6α.

The initiation of adaptive immunity relies on the performance of dendritic cells (DCs), which are specialized leukocytes with professional antigen presenting capabilities. As such, the molecular mechanisms safeguarding DC homeostasis are matter of intense research. Sensors of the unfolded protein response (UPR) of the endoplasmic reticulum, a three-pronged signaling pathway that maintains the fidelity of the cellular proteome, have emerged as regulators of DC biology. The archetypical example is the IRE1/XBP1s axis, which supports DC development and survival of the conventional type 1 DC (cDC1) subtype. However, the role of additional UPR sensors in DC biology, such as the ATF6α branch, has not been clearly elucidated. Even though Xbp1 is transcriptionally induced by ATF6α under ER stress, it is unclear if cDCs also co-opt the ATF6α branch in tissues. Here, we examine the role of ATF6α in cDC homeostasis in vivo and upon innate stimulation in vitro. In steady state, animals lacking ATF6α in CD11c+ cells (Itgax Cre x Atf6 fl/fl mice) display normal cDC frequencies in spleen, intestine, liver, and lung. Also, ATF6α deficient cDCs express normal levels of Xbp1 mRNA and additional UPR components. However, a reduction of lung monocytes is observed in Itgax Cre x Atf6 fl/fl conditional deficient animals suggesting that ATF6α may play a role in the biology of monocyte subsets. Notably, in settings of DC activation, ATF6α contributes to the production of IL-12 and IL-6 to inflammatory stimuli. Thus, although ATF6α may be dispensable for tissue cDC homeostasis in steady state, the transcription factor plays a role in the acquisition of selective immunogenic features by activated DCs.