Morphology Profile Research Articles

Germplasm characterization and SDS-PAGE analysis of caper (Capparis spinosa L.) from different provenances

BackgroundCapparis spinosa L. is a typical desert plant that is resistant to high temperatures and drought, and at the same time is rich in medicinal and food values. The objective of this study is to explore the variations in nutrient composition, morphological characteristics, and SDS-PAGE patterns of caper seeds from different provenances, aiming to provide insights for the selection of superior seed provenances.ResultsIn this experiment, there were significant differences in the morphological characteristics and major nutritional components of caper seeds from different provenances. Seeds from the YKL (Karayagaqi Township, Yining County) and YKG (G218, KashiTown, Yining County) regions were larger in size compared to seeds from other regions. Among the four measured nutritional components, crude fat had the highest content, especially in the YKL and YKG region. The results of correlation analysis showed that crude fat was negatively correlated with soluble sugar and soluble protein but significantly positively correlated with starch content. As longitude increased from east to west, the morphological characteristics gradually increased. Based on the principal component analysis of all the parameters of the seeds, the eight provenances could be classified into three groups. HM (Hami), TGS (S202, Gaochang District, Turpan), HYW (Wubao Town, Yizhou District, Hami), TQQ (Qiquanhu Town, Turpan), and TLF (Turpan) were a group with higher soluble protein, soluble sugar, and water content. YKL and YKG were in one group, which had larger seed grains with high crude fat and starch content. AKS (Aksu) was in a separate group. The protein fractions from seeds of eight regions were extracted using Osborne fractionation method, it was found that glutelin content was the highest, while albumin content was the lowest. After these proteins were analyzed by SDS-PAGE, the electrophoretic patterns showed that the protein molecular weights were relatively small, and there were differences in protein bands among different provenances.ConclusionAccording to the PCA results, the eight seed provenances could be divided into three groups. There were both geographically distant ones clustered into one group, and those close to each other were also divided into one group. There were differences in seed morphology, nutrient content and SDS-PAGE profiles among the different seed sources. This difference might be caused by a combination of geographic and climatic factors. In addition, YKL and YKG were roughly selected as good seed provenances, which provided a theoretical basis for the development of C. spinosa L. germplasm resources.

Temporally and spatially resolved molecular profiling in fingerprint analysis using indium vanadate nanosheets-assisted laser desorption ionization mass spectrometry

This study presents the first-ever synthesis of samarium-doped indium vanadate nanosheets (IVONSs:Sm) via microemulsion-mediated solvothermal method. The nanosheets were subsequently utilized as a nano-matrix in laser desorption/ionization mass spectrometry (LDI-MS). It was discovered that the as-synthesized IVONSs:Sm possessed the following advantages: improved mass spectrometry signal, minimal matrix-related background, and exceptional stability in negative-ion mode. These qualities overcame the limitations of conventional matrices and enabled the sensitive detection of small biomolecules such as fatty acids. The negative-ion LDI mechanism of IVONSs:Sm was examined through the implementation of density functional theory simulation. Using IVONSs:Sm-assisted LDI-MS, fingerprint recognitions based on morphology and chemical profiles of endogenous/exogenous compounds were also achieved. Notably, crucial characteristics such as the age of an individual’s fingerprints and their physical state could be assessed through the longitudinal monitoring of particular biomolecules (e.g., ascorbic acid, fatty acid) or the specific biomarker bilirubin glucuronide. Critical information pertinent to the identification of an individual would thus be facilitated by the analysis of the compounds underlying the fingerprint patterns.Graphical

Morphologic, phenotypic, and genotypic similarities between primary tumors and corresponding 3D cell cultures grown in a repeatable system—preliminary results

BackgroundThree-dimensional (3D) cell cultures are the new frontier for reproducing the tumor micro-environment in vitro. The aims of the study were (1) to establish primary 3D cell cultures from canine spontaneous neoplasms and (2) to demonstrate the morphological, phenotypic and genotypic similarities between the primary canine neoplasms and the corresponding 3D cultures, through the expression of tumor differentiation markers.ResultsSeven primary tumors were collected, including 4 carcinomas and 3 soft tissue sarcomas. 3D cell cultures reproduced the morphological features of the primary tumors and showed an overlapping immunophenotype of the primary epithelial tumors. Immunohistochemistry demonstrated the growth of stromal cells and macrophages admixed with the neoplastic epithelial component, reproducing the tumor microenvironment. Mesenchymal 3D cultures reproduced the immunophenotype of the primary tumor completely in 2 out of 3 examined cases while a discordant expression was documented for a single marker in one case. No single nucleotide variants or small indel were detected in TP53 or MDM2 genes, both in primary tumors and in 3D cell cultures specimens. In one sample, MDM2 amplicons were preferentially increased in number compared to TP53 ones, indicating amplification of MDM2, detectable both in the primary tumor and in the corresponding cell culture specimen.ConclusionHere we demonstrate a good cell morphology, phenotype and genetic profile overlap between primary tumors and the corresponding 3D cultures grown in a repeatable system.

The Development of Human Ex Vivo Models of Inflammatory Skin Conditions.

Traditional research in inflammatory dermatoses has relied on animal models and reconstructed human epidermis to study these conditions. However, these models are limited in replicating the complexity of real human skin and reproducing the intricate pathological changes in skin barrier components and lipid profiles. To address this gap, we developed experimental models that mimic various human inflammatory skin phenotypes. Human ex vivo skins were stimulated with various triggers, creating models for inflammation-induced angiogenesis, irritation response, and chronic T-cell activation. We assessed the alterations in skin morphology, cellular infiltrates, cytokine production, and epidermal lipidomic profiles. In the pro-angiogenesis model, we observed increased mast cell degranulation and elevated levels of angiogenic growth factors. Both the irritant and chronic inflammation models exhibited severe epidermal disruption, along with macrophage infiltration, leukocyte exocytosis, and heightened cytokine levels. Lipidomic analysis revealed minor changes in the pro-angiogenesis model, whereas the chronic inflammation and irritant models exhibited significant decreases in barrier essential ceramide subclasses and a shift toward shorter acyl chain lengths (<C18), indicating skin barrier instability. Additionally, the irritant and chronic inflammation models are responsive to immunosuppressants. These models hold promise for advancing scientific understanding and the development of therapeutic and skincare solutions for individuals afflicted by compromised skin conditions.

Platelets characteristics in patients with coronary artery disease and atrial fibrillation

The aim. To estimate the connections between platelet morphological characteristics and amino acids (AA) profile in patients with coronary artery disease (CAD) and atrial fibrillation (AF).&#x0D; Materials and methods. 300 patients were included in the study. They were divided into 3 groups: first (CAD) – 149 patients with CAD but without arrhythmias, second (CAD+AF) – 124 patients with CAD and AF paroxysm and control group – 27 patients without CAD and arrhythmias. Platelets AA level was detected by method of ion exchange liquid column chromatography. Total platelet count (PC), mean platelet volume (MPV), platelets distribution width (PDW), platelet-to-leukocyte (PLR) were obtained from common blood count.&#x0D; Results. Significant decline of PC and rise of MPV and PLR ratio was found in CAD patients and AF paroxysm in comparison with group with CAD without arrhythmia, p &lt; 0.05. Significant increase of isoleucine, leucine and decrease of threonine, serine, glycine, valine levels was found, changes in branched chain amino acids (rise of Isoleucine + Leucine / Valine ratio) and Glycine + Serine (decline Glycine + Serine sum) metabolism were revealed in CAD patients and AF paroxysm in comparison with group with CAD without arrhythmia, p &lt; 0.05. Phenylalanine / Tyrosine ratio was significantly elevated in CAD patients with and without AF, p &lt; 0.05.&#x0D; Taurine, serine, and glycine correlated with platelets morphological characteristics: PC (r = 0.714, r = 0.732 and r = 0.340 consequently), MPV (r = -0.724, r = -0.390 and r = -0.571 consequently), PDW (r = -0.666 and r = -0.364 consequently) and PLR (r = 0.586 and r = 0.648 consequently), p &lt; 0.05. Platelets glycine (AUC = 0.8760), valine (AUC = 0.8707), leucine (AUC = 0.8267) and threonine (AUC = 0.8213) levels are closely connected with AF paroxysm in CAD patients by ROC-analysis results, p &lt; 0.05.&#x0D; Platelets amino acids combinations: Glycine + 0.6 × Valine (AUC = 0.9987), Threonine – 3.6 × Leucine (AUC = 0.9040) were found in prediction AF paroxysm for CAD patients by ROC-analysis results, p &lt; 0.05.&#x0D; Conclusions. Connections between platelets morphology and AA profile violations in patients with CAD and AF paroxysm were found in our study.

Unraveling the potential of sonochemically achieved DyMnO3/Dy2O3 nanocomposites as highly efficient visible-light-driven photocatalysts in decolorization of organic contamination

In the present day, the widespread presence of lingering contaminants in ecosystems has prompted scientists to develop novel semiconductor nanoarchitectures that assist in photocatalytic reactions mediated by visible light. As a result, we propose to prepare a series of Dy-Mn-O based nano-catalysts using a sonochemical approach utilizing various ionic phases of surfactants as structure-directing agents. In this study, X-ray diffraction (XRD) and Rietveld refinement techniques were used to explore the fundamental effects of surfactants on the compositional-structural features of the materials. In terms of morphological profiles, DyMnO3/Dy2O3 (DM) nanostructures fabricated with Triton X-80 as a structure-directing agent showed the best uniformity with an acceptable size range between 14.14 and 52.35 nm. In the visible-light-driven photocatalytic domain, these nanocomposites provide high responsiveness based on their optical band gap value of 2.0 eV. According to our findings, two individual factors affect dye activity, namely dye type and concentration, which is why a high decomposition efficiency of 78.8% was obtained for 10 ppm Acid violet (AV) using DyMnO3/Dy2O3 nanocomposites after 120 min of exposure to visible light. Furthermore, radical quenching test confirmation confirmed the mechanistic behind the degradation process. This indicates that active species of O2•− and •OH may play a significant role in photocatalysis. As a result of repeated processes over three consecutive cycles, binary DyMnO3/Dy2O3 nanocomposites had an efficiency of 64.4% in removing dyes from the environment, indicating their high stability.

Biocompatibility of ABS and PLA Polymers with Dental Pulp Stem Cells Enhance Their Potential Biomedical Applications.

Polylactic Acid (PLA) and Acrylonitrile-Butadiene-Styrene (ABS) are commonly used polymers in 3D printing for biomedical applications. Dental Pulp Stem Cells (DPSCs) are an accessible and proliferative source of stem cells with significant differentiation potential. Limited knowledge exists regarding the biocompatibility and genetic safety of ABS and PLA when in contact with DPSCs. This study aimed to investigate the impact of PLA and ABS on the adhesion, proliferation, osteogenic differentiation, genetic stability, proteomics, and immunophenotypic profile of DPSCs. A total of three groups, 1- DPSC-control, 2- DPSC+ABS, and 3- DPSC+PLA, were used in in vitro experiments to evaluate cell morphology, proliferation, differentiation capabilities, genetic stability, proteomics (secretome), and immunophenotypic profiles regarding the interaction between DPSCs and polymers. Both ABS and PLA supported the adhesion and proliferation of DPSCs without exhibiting significant cytotoxic effects and maintaining the capacity for osteogenic differentiation. Genetic stability, proteomics, and immunophenotypic profiles were unaltered in DPSCs post-contact with these polymers, highlighting their biosafety. Our findings suggest that ABS and PLA are biocompatible with DPSCs and demonstrate potential in dental or orthopedic applications; the choice of the polymer will depend on the properties required in treatment. These promising results stimulate further studies to explore the potential therapeutic applications in vivo using prototyped polymers in personalized medicine.

Forming and Microstructure Evolution Mechanism of Asymmetric Molten Pool in Laser Cladding

This study aimed to investigate the impact of substrate tilt angle and different scanning methods on powder utilization, morphology profile parameters, and microstructure evolution in laser cladding single-track experiments. The results revealed several key findings. Firstly, as the substrate tilt angle increased, the laser spot projected on the tilted substrate transformed from circular to elliptical, and the substrate heat input decreased proportionally with the tilt angle. Secondly, regardless of the scanning method employed, powder utilization decreased with higher substrate tilt angles due to increased powder asymmetry, leading to more collisions and rebounds between the powder particles and the substrate. Moreover, the trends of morphology profile parameters in longitudinal upward and downward scanning methods were similar to the transverse scanning, except for the clad width, which exhibited an opposite trend. Notably, the longitudinal downward scanning method demonstrated superior stability and higher powder utilization. These findings offer valuable insights for optimizing laser cladding processes, with the ultimate goal of enhancing the quality and efficiency of the fabricated components.

Collective Synthesis of Sarpagine and Macroline Alkaloid-Inspired Compounds.

Design strategies that can access natural-product-like chemical space in an efficient manner may facilitate the discovery of biologically relevant compounds. We have employed a divergent intermediate strategy to construct an indole alkaloid-inspired compound collection derived from two different molecular design principles, i.e. biology-oriented synthesis and pseudo-natural products. The divergent intermediate was subjected to acid-catalyzed or newly discovered Sn-mediated conditions to selectively promote intramolecular C- or N-acylation, respectively. After further derivatization, a collection totalling 84 compounds representing four classes was obtained. Morphological profiling via the cell painting assay coupled with a subprofile analysis showed that compounds derived from different design principles have different bioactivity profiles. The subprofile analysis suggested that a pseudo-natural product class is enriched in modulators of tubulin, and subsequent assays led to the identification of compounds that suppress in vitro tubulin polymerization and mitotic progression.

Ossifying Fibromyxoid Tumor with Atypical Features: A Case Report and Review of Literature

Abstract Introduction/Objective Ossifying fibromyxoid tumor (OFMT) is a soft tissue neoplasm usually behaving in a benign fashion, however, it can rarely show atypical or malignant features. It most frequently occurs within the extremities and trunk. Although OFMTs are translocation-associated tumors, and very commonly associated with recurrent gene rearrangements involving the PHF1 gene, their morphologic and immunophenotypic profile can overlap with other soft tissue tumors posing a diagnostic challenge. Methods/Case Report Herein we present a case of a 23-year-old woman who had an excision of a subcutaneous shoulder mass. Grossly, the 5.3 cm mass had a thin translucent capsule and a pink, soft-cut surface with scattered calcifications. Microscopic examination revealed a well-circumscribed lesion composed of plump spindle cells with uniform oval nuclei arranged in short fascicles within a mildly collagenous background. There was prominent hemangiopericytoma-like vasculature and foci of ossification. The tumor cells contained moderate cytoplasm and showed a mild degree of nuclear atypia. Mitotic activity was up to 6 per 10 high-power fields. Immunohistochemical stains showed tumor cells are positive for Desmin, SMA, AE1/AE3 (patchy), TLE, and SATB2, while negative for CD31, CD34, S-100, C-kit, DOG-1, CD117, HMB-45, Melan-A, ALK-1, SOX10, Inhibin, and STAT6. Ki-67 proliferation index was low (1-2%). Fluorescence in situ hybridization (FISH) testing was negative for SYT and MDM2 gene amplification. FISH testing for PHF1 gene (using experimental probes) showed a break apart signal in a subset (~ 25%) of tumor cells, which is equivocal but insufficient to establish clonal interchromosomal translocation involving PHF1. Results (if a Case Study enter NA) NA Conclusion There is a histologic spectrum and morphologic overlap of OFMTs with several other soft tissue neoplasms. OFMT with atypical features (increased cellularity and mitotic activity), as in this case, may harbor the unpredictable biologic potential for recurrence and metastasis. Additional studies are required to discover a morphologic, immunohistochemical, and possibly molecular pattern that will help to differentiate OFMT subtypes.

Bacillus mojavensis isolated from aguamiel and its potential as a probiotic bacterium

BackgroundMillenary fermented beverages are a source of industrially important microorganisms. Aguamiel and pulque are traditional Mexican beverages of pre-Hispanic origin, with a microbial diversity that contributes to the different fermentations (lactic, alcoholic and acetic). The aim of this research was to characterize the Bacillus mojavensis (BF2A1) strain isolated from aguamiel and determine its probiotic potential. The strain was identified through Mass Spectrometry (MS), molecular techniques, as well as morphological and biochemical profiling. The probiotic activity of the BF2A1 strain and its response during the gastric simulation was determined. ResultsThe strain BF2A1 is a Gram-positive, spore-forming bacillus, positive for catalase, gamma-hemolysis, citrate, ornithine, grows at 7.5% NaCl, and acetoin, but negative for motility, indole, and methyl red. Its taxonomic identity was determined as B. mojavensis both by MALDI-TOF MS and sequencing of 16S rDNA. Its probiotic potential was demonstrated as BF2A1 was tolerant to pH 2 (OD620nm 0.289 ± 0.012), 0.3% bile salts (OD620nm 0.103 ± 0.089), 8% NaCl (OD620nm 0.254 ± 0.096), and 1% lysozyme (OD620nm 1.342 ± 0.078) compared to the probiotic strain Lactobacillus leichmannii ATCC 7830TM (Laclei). The antagonistic effect of BF2A1 against Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC25923), and Candida albicans (ATCC60193) showed 25.5%, 8% and, 65% inhibitory growing effect, respectively. BF2A1 in the gastric simulation showed only a reduction of 1–2 log CFU/mL and showed after the intestinal phase a survival rate of 84.4% as compared to the control strains. ConclusionsThis study shows that BF2A1 isolated from aguamiel is a bacterium with probiotic properties that can be used in different areas of Biotechnology.How to cite: Martínez-Ortiz V, Trujillo-López MA, El-Kassis EG, et al. Bacillus mojavensis isolated from aguamiel and its potential as a probiotic bacterium. Electron J Biotechnol 2024;67. https://doi.org/10.1016/j.ejbt.2023.11.002.

Label-Free High Dimensional Single Cell Morphological Profiling of Different Hematological Malignancies By Ghost Cytometry

Characterizing the heterogeneity in human leukemias is critical for understanding mechanisms of disease onset, uncovering biomarkers for disease diagnosis, and guiding research into novel treatment approaches. While extensive research focus has been placed on characterizing the molecular heterogeneity in leukemia by modern 'omics technologies, comparatively less emphasis has been placed on studying the diversity in holistic morphological changes that occur in single cells in the context of disease. Here, we report on a novel approach to understanding and characterizing different hematological malignancies using ghost cytometry, a recently developed high-content flow cytometric method that leverages high-speed, artificial intelligence (AI) driven morphological characterization and analysis of single cells. We performed morphometric profiling on human bone marrow samples from patients with T and B-cell acute lymphoblastic leukemia (ALL, n=5), acute myeloid leukemia (AML, n=5), multiple myeloma (MM, n=5) and healthy bone marrow mononuclear cell (BM-MNC) controls (n=6) (Figure 1). Morphological profiles from individual cells were analyzed by unsupervised machine learning (universal manifold approximation and projection, UMAP) and compared within disease subsets and against controls. We identified disease-specific cell populations from a mixture of healthy control and disease samples without any labeling, based on completely label-free information (Figure 2). For validation of the specific cell subsets, we overlaid known cell surface markers (e.g. CD45 for blast cells, CD10/CD19 for B-ALL, CD33/CD34 for AML, CD38/CD138 for MM). The intensity of CD45 was dimmer and disease-specific markers were highly expressed in the disease-specific cell population. We also identified unique cell populations across blood cell populations that were distinct from the healthy BM-MNC controls. We also analyzed three human myeloma cell lines (AMO1, KMS, and L363) for comparison to the primary MM samples. In the UMAP analysis using LF-GC data, in all three types of cell lines, Bortezomib-resistant cell lines exhibited different distributions from the control parental cell lines. In addition, we observed distinct morphological profiles between primary myeloma samples and myeloma cell lines. Here we present a novel cell characterization approach for human hematological diseases that leverages AI-based, label-free, high-speed morphological characterization of single cells. We demonstrate that the approach can be used to identify subtle morphological differences in blast cells from a range of blood cancer subtypes. Application of morphological profiling and AI can be used to identify measurable, disease-related changes in these diseases that have diagnostic, drug screening, and therapeutic monitoring potential. Keywords: flow cytometry, acute leukemia, multiple myeloma, label-free cell analysis, biomarker, liquid biopsy, ghost cytometry, artificial intelligence, machine learning Figure 1. Research design overview. Bone marrow samples from patients and non-diseased controls will be analyzed by Ghost Cytometry to assess disease by conventional markers and identify novel morphometric subpopulations. Morphological information is acquired as label-free GMI signals without any labeling. The label-free GMI information is transformed into two-dimensional space by UMAP dimensionality reduction method to identify subpopulations of interest. Both samples are stained with known cell surface markers for validation of unsupervised classification. Figure 2. Identification of disease-specific cell subpopulations by unsupervised approach. Using an unsupervised machine learning technique called UMAP, label-free morphologically distinct cellular subpopulations can be identified in B-ALL, AML, MM when compared to healthy donor controls (left panels, inside the dotted circles). For validation of the subpopulations, known cell surface markers (CD45 for blast cells, CD10/CD19 for B-ALL, CD33/CD34 for AML, CD38/CD138 for MM) and cell trace marker (CellTracker) were overlaid. The intensity increases monotonically through the colormaps. The intensity of CD45 was dimmer and disease-specific markers were highly expressed in the disease-specific cell population.

Morphologic and Genetic Features of Unclassifiable Chronic Myeloid Neoplasms: Five-Year Experience of a Canadian Provincial Referral Center

Introduction: The challenge of diagnosing and definitively classifying myeloproliferative neoplasms (MPNs) and myelodysplastic/myeloproliferative neoplasms (MDS/MPNs) remains a conundrum in the field of hematology. While the World Health Organization has outlined diagnostic criteria for various categories, there remain cases that elude specific classification, falling under the umbrella of MPN, unclassifiable (MPN-U) and MDS/MPN, unclassifiable (MDS/MPN-U). In this study, we aimed to compare the morphological and genetic profiles of cases classified as MPN-U or MDS/MPN-U against more definitively classified cases (including chronic myelomonocytic leukemia (CMML), essential thrombocythemia (ET), MDS/MPN with ring sideroblasts and thrombocytosis (MDS/MPN-RS-T), and primary myelofibrosis (PMF), including prefibrotic PMF (pre-PMF)). Methods: Morphologically unclassifiable myeloid neoplasm cases referred for pathologist review to our laboratory in the largest tertiary care center in British Columbia, Canada were identified through pathology database query over a five-year period. Bone marrow materials were reviewed and reanalyzed with standardized collection of qualitative and semiquantitative data by two hematopathologists. Morphological parameters studied included the myeloid to erythroid ratio, bone marrow cellularity, lineage dysplasia, megakaryocyte histotopography, bone marrow blast frequency, fibrotic changes, and bone marrow vascularity, among others. Clinical data, cytogenetic findings, and results of a targeted sequencing panel assessing a set of approximately 50 genes recurrently mutated in myeloid malignancies were collected by electronic medical record review. A cohort of contemporarily collected myeloid neoplasms with definitive pathologic diagnoses was reviewed for comparison. Statistical analyses were performed using python3. Results: The cohort comprised 29 cases classified as MPN-U, 12 cases of MDS/MPN-U, and 5 cases each of ET, PMF, pre-PMF, CMML, and MDS/MPN-RS-T. MPN-U cases were further subcategorized based on the apparent reason for difficulty in classification, including: overlapping features with another MPN; late stage/overt fibrosis; early stage without fully-developed morphologic features; and coexisting atypical morphologic findings. MPN-U with overlapping features had highly variable reticulin fibrosis, while, as expected, late stage entities had a high degree of reticulin fibrosis and early stage entities showed less overt fibrosis (Figure, Left, demonstrating select morphologic features of evaluated chronic myeloid neoplasms). MPN-U as a whole appeared to show less prominent fibrosis-associated findings than MDS/MPN-U and to, in general, show a greater degree of neovascularization, similar to changes observed in cases with PMF. Splicing factor and epigenetic modifier mutations appeared to be overrepresented in MDS/MPN-U relative to MPN-U, notably excluding MPN-U with atypical morphologic features (Figure, Right, demonstrating the distribution of pathogenic variants identified in evaluated myeloid neoplasms; results are indicated as presence (1) or absence (0) of a variant for each gene). Notably, no CALR mutations were identified in the MDS/MPN-U category or early stage / atypical morphology MPN-U, though they were enriched in overtly fibrotic MPN-U entities. Discussion/Conclusion: In this cohort we observed, as expected, significant overlap in cases classified as MPN-U and MDS/MPN-U with better delineated entities, though patterns of morphologic change and mutational spectra suggest potential directions for further investigation of additional criteria that may guide accurate classification of these neoplasms. These findings reflect the heterogeneity of MPNs described in the literature and the complexity of their genetic underpinnings, particularly in the context of concurrent dyshematopoiesis; suggest enhanced bone marrow vascularization may be an informative differential diagnostic criterion in evaluation of these neoplasms, though further study is required; and emphasize the need for broader genetic testing and perhaps further subtyping of MPN-U cases for a more nuanced analytical approaches for these disorders.

Morphology and reproductivity profiling of male Senduro goats based on age differences

Senduro goats, a local breed of meat and dairy goats from Indonesia, are recognized for their significance in improving goat breeding and preserving valuable genetic resources. However, limited information exists regarding the reproductive physiology of Senduro goats, which poses challenges to the development of breeding programs and the preservation of genetic resources. This study aimed to investigate the morphological and reproductive profiles of male Senduro goats at different ages, focusing on morphological characteristics, mating behavior, and sperm quality. Morphological characteristics are assessed through body length measurements, while mating behavior serves as an indicator of reproductive behavior. Macroscopic evaluations of sperm quality include assessments of color, viscosity, pH, and volume, while microscopic examinations encompass motility (mass and individual), viability, and spermatozoa membrane integrity. The results showed morphological similarities between juvenile and adult samples, with their testicular size being the only significant difference. Based on macroscopic and microscopic examinations, no significant differences were found between groups. From the results it was concluded that there were no distinct differences in morphological characteristics, mating behavior, and sperm quality between male Senduro goats in the juvenile and adult stages.

Discovery of Molecular Glue Degraders via Isogenic Morphological Profiling.

Molecular glue degraders (MGDs) are small molecules that degrade proteins of interest via the ubiquitin-proteasome system. While MGDs were historically discovered serendipitously, approaches for MGD discovery now include cell-viability-based drug screens or data mining of public transcriptomics and drug response datasets. These approaches, however, have target spaces restricted to the essential proteins. Here we develop a high-throughput workflow for MGD discovery that also reaches the nonessential proteome. This workflow begins with the rapid synthesis of a compound library by sulfur(VI) fluoride exchange chemistry coupled to a morphological profiling assay in isogenic cell lines that vary in levels of the E3 ligase CRBN. By comparing the morphological changes induced by compound treatment across the isogenic cell lines, we were able to identify FL2-14 as a CRBN-dependent MGD targeting the nonessential protein GSPT2. We envision that this workflow would contribute to the discovery and characterization of MGDs that target a wider range of proteins.

Extranodal NK/T-Cell Lymphoma Predominantly Composed of Anaplastic Cells: A Frequently Misdiagnosed and Highly Aggressive Variant.

Extranodal NK/T-cell lymphoma (ENKTL) is a non-Hodgkin lymphoma associated with the Epstein-Barr virus that primarily affects individuals in East Asia and indigenous populations in Central and South America. Morphologically, ENKTL typically consists of medium-sized cells or a combination of small and large cells. This report presents 10 cases characterized by predominantly anaplastic cells with diffuse expression of CD30, resembling anaplastic lymphoma kinase-negative anaplastic large cell lymphoma (ALK-negative ALCL) and demonstrating highly aggressive behavior. The cohort included 9 males and 1 female, ranging in age from 29 to 65 years (median age: 47y). Eight patients presented with nasal disease, while 2 had non-nasal disease. Five patients had stage I/II disease, and the remaining 5 had stage III/IV disease. Morphologically, necrosis was observed in 9 cases, angiocentric-angiodestructive growth in 3 cases, and pseudoepitheliomatous hyperplasia in 2 cases. Anaplastic cells predominated in all cases, with some displaying eccentric, horseshoe-shaped, or kidney-shaped nuclei (referred to as "Hallmark" cells). The morphology profile was monomorphic in 3 cases and polymorphic in 7 cases. Immunohistochemically, all cases tested positive for cytotoxic granule markers (TIA1 and granzymeB) and Epstein-Barr virus-encoded RNA. Cytoplasmic expression of CD3ε and CD56 was observed in 9 of 10 cases. Interestingly, most cases (7 of 8) exhibited variable expression of MuM1, ranging from 10% to 90%. All cases showed diffuse positivity for CD30 but were negative for ALK, resulting in 3 cases being initially misdiagnosed as ALK-negative ALCL. Compared with nonanaplastic cases, anaplastic cells predominant ENKTL had a significantly higher frequency of "B" symptoms, bone marrow involvement, hemophagocytic lymphohistiocytosis, and higher Ki67 proliferative index. These findings provide valuable information for pathologists, expanding their understanding of the cytologic spectrum of ENKTL. This rare variant of ENKTL, characterized by the predominance of anaplastic cells and diffuse CD30 expression, exhibits high aggressiveness and should be differentiated from ALK-negative ALCL. Awareness of this uncommon variant is crucial in preventing misdiagnosis and ensuring the timely initiation of therapy.

Cobalt and Titanium Alleviate the Methylglyoxal-Induced Oxidative Stress in Pennisetum divisum Seedlings under Saline Conditions.

Salinity is considered to be a global problem and a severe danger to modern agriculture since it negatively impacts plants' growth and development at both cellular- and whole-plant level. However, cobalt (Co) and titanium (Ti), multifunctional non-essential micro-elements, play a crucial role in improving plant growth and development under salinity stress. In the current study, Co and Ti impact on the morphological, biochemical, nutritional, and metabolic profile of Pennisetum divisum plants under three salinity levels which were assessed. Two concentrations of Co (Co-1; 15.0 mg/L and Co-2; 25.0 mg/L), and two concentrations of Ti (Ti-1; 50.0 mg/L and Ti-2; 100.0 mg/L) were applied as foliar application to the P. divisum plants under salinity (S1; 200 mM, S2; 500 mM, and S3; 1000 mM) stress. The results revealed that various morphological, biochemical, and metabolic processes were drastically impacted by the salinity-induced methylglyoxal (MG) stress. The excessive accumulation of salt ions, including Na+ (1.24- and 1.21-fold), and Cl- (1.53- and 1.15-fold) in leaves and roots of P. divisum, resulted in the higher production of MG (2.77- and 2.95-fold) in leaves and roots under severe (1000 mM) salinity stress, respectively. However, Ti-treated leaves showed a significant reduction in ionic imbalance and MG concentrations, whereas considerable improvement was shown in K+ and Ca2+ under salinity stress, and Co treatment showed downregulation of MG content (26, 16, and 14%) and improved the antioxidant activity, such as a reduction in glutathione (GSH), oxidized glutathione (GSSG), Glutathione reductase (GR), Glyoxalase I (Gly I), and Glyoxalase II (Gly II) by up to 1.13-, 1.35-, 3.75-, 2.08-, and 1.68-fold under severe salinity stress in P. divisum roots. Furthermore, MG-induced stress negatively impacted the metabolic profile and antioxidants activity of P. divisum's root and leaves; however, Co and Ti treatment considerably improved the biochemical processes and metabolic profile in both underground and aerial parts of the studied plants. Collectively, the results depicted that Co treatment showed significant results in roots and Ti treatment presented considerable changes in leaves of P. divism under salinity stress.

Insights into the functional properties of a natural free amino acid mix: Effect on growth performance, nutrient metabolism, and immune response in a carnivorous fish, Asian seabass (Lates calcarifer)

Dietary supplements containing a functional feed additive have been shown to be beneficial to fish and shellfish aquaculture. However, the functional properties of aquafeed formulations have rarely been reported in fish. This study aimed to investigate the effects of natural free amino acid mix (FAAM) supplementation as a functional solution on the growth performance and nutrient utilization in a carnivorous fish, Asian seabass (Lates calcarifer). Five isonitrogenous and isolipidic diets were prepared with graded supplementation levels of FAAM at 0 % (control group), 0.25 %, 0.50 %, 0.75 %, and 1.0 %, denoted as FAAM0, FAAM0.25, FAAM0.5, FAAM0.75, and FAAM1.0, respectively. The experimental fish were fed different dietary FAAM supplementations to apparent satiation twice daily for eight weeks. Significant improvements were observed in the growth performance of fish among the five groups (P < 0.05). Fish fed with FAAM0.75 displayed significantly increased activities of lysozyme, myeloperoxidase, catalase, and glutathione peroxidase (P < 0.05). The activities of digestive enzymes, including amylase, protease, and lipase, were enhanced by the supplementation of FAAM in the feed (P < 0.05), especially for the groups that contained more than 0.5 % FAAM in the feed. Furthermore, the morphological profile of the intestinal tract, including the mucosal fold height, width, thickness, and goblet cell, increased in fish fed with FAAM at 1.0 % (P < 0.05). Moreover, FAAM supplementation in diets not only modulated the expression of immune-related genes (glutathione peroxidase (GPx), complement (C)3, C4, and C-reactive protein) in the liver but also positively impacted the growth-ralated genes, including growth hormone (GH), GH receptor (GHR), insulin-like growth factor I (IGF-I), and IGF-II. In addition, the amounts of monounsaturated fatty acids (mainly oleic acid (C18:1n9c)) and polyunsaturated fatty acids-especially γ-linolenic acid (C18:3 n6) and α-linolenic acid (C18:3n3)-increased in fish fed with diets containing FAAMs (P < 0.05). Interestingly, the diets supplemented with FAAMs also had a positive effect on the economic indices in terms of revenue-to-cost ratios. These findings provide a scientific basis for the application of FAAMs as a functional solution that can be used in feed formulations for Asian seabass.

Viability Analysis of Biomphalaria glabrata Hemocytes During Schistosoma mansoni Infection And Glyphosate-Based Herbicide Exposure

Pesticides are chemical agents that have a range of harmful effects on human health and the environment. They can cause drastic changes in natural communities, such as macroinvertebrates, plankton and fish. Snails, including Biomphalaria glabrata, are often present in aquatic communities and have multiple roles in limnic ecosystems. B. glabrata is an intermediate host of several species of helminths of medical and veterinary importance, such as Schistosoma mansoni, the etiological agent of schistosomiasis. The original Roundup® herbicide can affect snails and directly affects S. mansoni hemocytes, cells that act in the snails’ immune defense. Here we analyzed the effect of herbicide exposure on B. glabrata hemocytes divided into four groups: control group, infected-only group, treated-only group and infected+treated group. For this, flow cytometry and Neubauer-chamber counting were used to determine the morphology, viability and lectin expression profiles of hemocytes. We observed that the group infected by S. mansoni and treated with herbicide had a higher concentration of dead hemocytes in relation to the other groups. The treated group showed similar results to the control group, suggesting that the herbicide (Roundup™) alone does not interfere with the snails’ immune system. Regarding cellular-morphological-characterization analysis, hyalinocytes were the cells most commonly found in all groups studied. These findings suggest that S. mansoni infection and exposure to pesticides directly the immune system of the snails, stimulating the production of hemocytes, especially hyalinocytes, which have a high phagocytic power to quell the infection, but with toxic effects on the snail.

EPCO-34. IMAGE-BASED PHENOTYPIC HIGH-CONTENT CRISPR-CAS9 SCREEN FOR CHARACTERIZING CANCER DRIVERS IN PEDIATRIC HIGH-GRADE GLIOMA

Abstract The genomic landscape of pediatric high-grade glioma is characterized by a high level of aneuploidy but few focal amplifications or homozygous deletions. Cancer aneuploidies are the principal source of tumor heterogeneity driving tumor growth and therapy resistance. Here, we deployed an image-based phenotypic high-content CRISPR-Cas9 screen to identify aneuploidy-associated pediatric glioma driver genes. Through integrated computational analyses combining whole-genome sequencing datasets from 106 pediatric gliomas and 81 canine gliomas, we identified a set of 460 genes present in regions of aneuploidy in both diseases. We designed a custom guideRNA library to systemically knockout the 460 gene set using an arrayed CRISPR-Cas9 knockout screen in two pediatric high-grade glioma cell lines. By deploying 72 of 384-well plates on automated confocal Opera Phenix High-Content Screening System and dyes staining cell membrane, nucleus, and apoptotic cells, we established 5 million live-cell images across three timepoints (day 7, 8, 9 post-transduction). Each gene was randomized to 32 randomized wells across four 384-well plates to gather robust morphological profiles. Through image-based nucleus and cell segmentation workflow, we characterized 50 million single cells across 27,648 wells over three time points. Using semi-supervised machine learning methods, we derived 1,050 cell morphological features to characterize knockout phenotypes, e.g., viability, growth, and morphology (size, shape, texture of nucleus and cells) across cells. The top ranked genes were then linked to oncogenes and tumor suppressors based on pathway and ontology analysis as well as functional in vitro and in silico validation using Pediatric Cancer Dependency Map. We find convergence of the most impactful molecular abnormalities - based on their knockout phenotypes - on candidate signaling pathways for the development of new drugs and repurposing of existing drugs for pediatric high-grade glioma.