Tissue Localization Research Articles

Applications of single‑cell omics and spatial transcriptomics technologies in gastric cancer (Review).

Gastric cancer (GC) is a prominent contributor to global cancer-related mortalities, and a deeper understanding of its molecular characteristics and tumor heterogeneity is required. Single-cell omics and spatial transcriptomics (ST) technologies have revolutionized cancer research by enabling the exploration of cellular heterogeneity and molecular landscapes at the single-cell level. In the present review, an overview of the advancements in single-cell omics and ST technologies and their applications in GC research is provided. Firstly, multiple single-cell omics and ST methods are discussed, highlighting their ability to offer unique insights into gene expression, genetic alterations, epigenomic modifications, protein expression patterns and cellular location in tissues. Furthermore, a summary is provided of key findings from previous research on single-cell omics and ST methods used in GC, which have provided valuable insights into genetic alterations, tumor diagnosis and prognosis, tumor microenvironment analysis, and treatment response. In summary, the application of single-cell omics and ST technologies has revealed the levels of cellular heterogeneity and the molecular characteristics of GC, and holds promise for improving diagnostics, personalized treatments and patient outcomes in GC.

C. elegans insulin-like peptides

Insulin-like peptides are a group of hormones crucial for regulating metabolism, growth, and development in animals. Invertebrates, such as C. elegans, have been instrumental in understanding the molecular mechanisms of insulin-like peptides. Here, we review the 40 insulin-like peptide genes encoded in the C. elegans genome. Despite the large number, there is only one C. elegans insulin-like peptide receptor, called DAF-2. The insulin and insulin-like growth factor signaling (IIS) pathway is evolutionarily conserved from worms to humans. Thus C. elegans provides an excellent model to understand how these insulin-like peptides function. C. elegans is unique in that it possesses insulin-like peptides that have antagonistic properties, unlike all human insulin-like peptides, which are agonists. This review provides an overview of the current literature on C. elegans insulin-like peptide structures, processing, tissue localization, and regulation. We will also provide examples of insulin-like peptide signaling in C. elegans during growth, development, germline development, learning/memory, and longevity.

21. VARIATION OF MICROBIOTA DEPENDING ON THE LOCATION OF COLORECTAL CANCER

Abstract Introduction Colorectal cancer (CRC) growth may be determined by various factors, with gut microbiota being one of them. The aim of this study is to Identify the microbiota differences in tumoral tissue (TT), non-tumoral tissue (NTT) and feces depending on CRC localization. Methods Prospective analysis was conducted on patients with CRC who underwent colorectal surgery in 2021 and 2022. Three samples were collected from each patient: feces, tissue from tumor (TT), and tissue from healthy colon (NTT). The study employed 16S rRNA massive sequencing techniques, followed by bioinformatics analyses. Results A total of 23 patients, comprising 18 males and 5 females, were included in the study. Tumor distribution revealed 12 cases in the right colon, 7 in the left colon, and 4 in the rectum. Significant differences were observed in beta diversity between the microbiota of feces and both types of tissue (TT and NTT). However, no differences in beta diversity were noted between TT and NTT. NTT exhibited a predominance of the phylum Actinobacteriota, while Bacteroidetes was the main phylum in feces. TT was characterized by a higher proportion of Fusobacteriota, with the genera Fusobacteria and Streptococcus being predominant compared to feces and the genus Fusobacteria compared to NTT. When considering tumor location, notable differences emerged. There was greater microbiota diversity in the right colon compared to the left colon and rectum. Phyla Clostridiales, Bifidobacteriales, Acidaminococcales, and Vibrionales were more abundant in the right colon, while Staphylococcales were more prevalent in the left colon. Conclusion The observed variations in microbiota based on tissue type and location imply its potential involvement in colorectal cancer (CRC) pathogenesis.

IGF2BP2-m6A-circMMP9 axis recruits ETS1 to promote TRIM59 transcription in laryngeal squamous cell carcinoma

Laryngeal squamous cell carcinoma (LSCC) is a common malignancy of the head and neck. Recently, circular RNA (circRNA) has been studied extensively in multisystem diseases. However, there are few research on biological functions and molecular mechanisms of circRNAs in LSCC. CircRNA array was used to detect the differentially expressed circRNAs. Kaplan–Meier and cox regression analysis were used to identify survival based on circMMP9. The qRT-PCR, RNase R treatment, sanger sequencing and in situ hybridization were used to verify circMMP9 expression, characteristics and localization in LSCC tissues and cells. Functionally, colony formation, MTS, transwell and in vivo assays were proceeded to detect the biological function of circMMP9 in LSCC progression. The RNA-seq was conducted to identify the molecular targets of circMMP9. Mechanically, MeRIP, RNA Immunoprecipitation (RIP), RNA pulldown, Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays were carried on to verify the regulatory mechanism of circMMP9. CircMMP9 was discovered upregulated in LSCC tissues and cells, and high level of circMMP9 was associated with poor prognosis, low degree of pathological grading, high TNM stage and lymph node metastasis of LSCC. CircMMP9 knockdown prevented LSCC progression both in vitro and in vivo, whereas, circMMP9 overexpression had the opposite effect. CircMMP9 was stabilized by IGF2BP2 in m6A-dependent manner. TRIM59 was identified as downstream target of circMMP9. CircMMP9 recruited ETS1 to stimulate TRIM59 transcription. Moreover, TRIM59 accelerated LSCC progression via activating the PI3K/AKT signal pathway. Our findings offered a unique regulatory mechanism for circMMP9 in LSCC, as well as a novel proof that circMMP9 may be utilize as a diagnostic marker and therapeutic target for LSCC patients.

Energy-guided diffusion model for CBCT-to-CT synthesis

Cone Beam Computed Tomography (CBCT) plays a crucial role in Image-Guided Radiation Therapy (IGRT), providing essential assurance of accuracy in radiation treatment by monitoring changes in anatomical structures during the treatment process. However, CBCT images often face interference from scatter noise and artifacts, posing a significant challenge when relying solely on CBCT for precise dose calculation and accurate tissue localization. There is an urgent need to enhance the quality of CBCT images, enabling a more practical application in IGRT. This study introduces EGDiff, a novel framework based on the diffusion model, designed to address the challenges posed by scatter noise and artifacts in CBCT images. In our approach, we employ a forward diffusion process by adding Gaussian noise to CT images, followed by a reverse denoising process using ResUNet with an attention mechanism to predict noise intensity, ultimately synthesizing CBCT-to-CT images. Additionally, we design an energy-guided function to retain domain-independent features and discard domain-specific features during the denoising process, enhancing the effectiveness of CBCT-CT generation. We conduct numerous experiments on the thorax dataset and pancreas dataset. The results demonstrate that EGDiff performs better on the thoracic tumor dataset with SSIM of 0.850, MAE of 26.87 HU, PSNR of 19.83 dB, and NCC of 0.874. EGDiff outperforms SoTA CBCT-to-CT synthesis methods on the pancreas dataset with SSIM of 0.754, MAE of 32.19 HU, PSNR of 19.35 dB, and NCC of 0.846. By improving the accuracy and reliability of CBCT images, EGDiff can enhance the precision of radiation therapy, minimize radiation exposure to healthy tissues, and ultimately contribute to more effective and personalized cancer treatment strategies.

Plasma fibroblast activation protein is decreased in acute heart failure despite cardiac tissue upregulation

BackgroundCardiac fibroblast activation protein (FAP) has an emerging role in heart failure (HF). A paradoxical reduction in its levels in pathological conditions associated with acute processes has been observed. We aimed to identify FAP cardiac tissue expression and its relationship with the main cardiac fibrosis-related signaling pathways, and to compare plasma FAP levels in acute and chronic HF patients.MethodsTranscriptomic changes were assessed via mRNA/ncRNA-seq in left ventricle tissue from HF patients (n = 57) and controls (n = 10). Western blotting and immunohistochemistry were used to explore FAP protein levels and localization in cardiac tissue. ELISA was performed to examine plasma FAP levels in acute HF (n = 48), chronic HF (n = 15) and control samples (n = 7).ResultsFAP overexpression in cardiac tissue is related to the expression of molecules directly involved in cardiac fibrosis, such as POSTN, THBS4, MFAP5, COL1A2 and COL3A1 (P < 0.001), and is directly and inversely related to pro- and antifibrotic microRNAs, respectively. The observed FAP overexpression is not reflected in plasma. Circulating FAP levels were lower in acute HF patients than in controls (P < 0.05), while chronic HF patients did not show significant changes. The clinical variables analyzed, such as functional class or etiology, do not affect plasma FAP concentrations.ConclusionsWe determined that in HF cardiac tissue, FAP is related to the main cardiac fibrosis signaling pathways as well as to pro- and antifibrotic microRNAs. Additionally, an acute phase of HF decreases plasma FAP levels despite the upregulation observed in cardiac tissue and regardless of other clinical conditions.Graphical abstract

Cover Image:

Pericycle‐localized OsK5.2 in Rice Root: Rice is a staple food for half of the world's population, but rice grains have low calcium (Ca) concentrations. Wang et al. (Pages 252‐264) report that a mutant with loss of function of OsK5.2 showed higher Ca accumulation in both the grain and straw. OsK5.2 localizes to the root pericycle and participates in the root‐to‐shoot translocation of Ca. The cover shows tissue localization of OsK5.2 (red) detected by immunostaining and root cell walls in light blue.

Canine soft tissue sarcomas: the expression of RUNX2 and karyopherin alpha-2 in extraskeletal (soft tissues) and skeletal osteosarcomas.

Extraskeletal osteosarcoma (EOS) is a malignant tumor producing bone matrix and/or chondroid material, without direct attachment to bone or periosteum. In humans and dogs, EOS is highly infiltrating, rapidly growing, often characterized by osteoid deposition and variable ossification, similar to primary skeletal osteosarcoma (SOS). In dogs, EOS arises from visceral and soft tissue locations, occasionally in trauma or foreign body sites, or in granulomas. Few data are currently available on the phenotype of these tumors. The present study aims to assess the expression RUNX2 and Karyopherin alpha-2 in EOS, comparing it with SOS and the data available from the human counterpart. Seventeen cases of canine osteosarcoma (13 EOS and 4 SOS) were retrospectively selected and submitted to immunohistochemistry for RUNX2 and Karyopherin alpha-2. Our results showed that, in EOS, RUNX2 is expressed in a mean of 73.07 ± 5.36 neoplastic cell nuclei, in face of a mean 36.15 ± 6.25 of Karyopherin alpha-2 positive nuclei. Osteoclasts, when present, were negative for both markers. No correlation was observed among the two markers (p > 0.05), nor statistically significant difference in quantitative expression was assessed comparing EOS and SOS groups. RUNX2 is expressed in canine EOS similarly to SOS and could be used as a diagnostic marker in a larger panel. Karyopherin alpha-2 is expressed in canine EOS and SOS similarly to human SOS and could be validated in future studies as an additional diagnostic marker. Further studies should be planned to evaluate the expression of these proteins as prognostic predictive parameters.

Epileptic tissue localization using graph-based networks in the high frequency oscillation range of intracranial electroencephalography

PurposeHigh frequency oscillations (HFOs) are an emerging biomarker of epilepsy. However, very few studies have investigated the functional connectivity of interictal iEEG signals in the frequency range of HFOs. Here, we study the corresponding functional networks using graph theory, and we assess their predictive value for automatic electrode classification in a cohort of 20 drug resistant patients. MethodsCoherence-based connectivity analysis was performed on the iEEG recordings, and six different local graph measures were computed in both sub-bands of the HFO frequency range (80–250 Hz and 250–500 Hz). Correlation analysis was implemented between the local graph measures and the ripple and fast ripple rates. Finally, the WEKA software was employed for training and testing different predictive models on the aforementioned local graph measures. ResultsThe ripple rate was significantly correlated with five out of six local graph measures in the functional network. For fast ripples, their rate was also significantly (but negatively) correlated with most of the local metrics. The results from WEKA showed that the Logistic Regression algorithm was able to classify highly HFO-contaminated electrodes with an accuracy of 82.5 % for ripples and 75.4 % for fast ripples. ConclusionFunctional connectivity networks in the HFO band could represent an alternative to the direct use of distinct HFO events, while also providing important insights about hub epileptic areas that can represent possible surgical targets. Automatic electrode classification through FC-based classifiers can help bypass the burden of manual HFO annotation, providing at the same time similar amount of information about the epileptic tissue.

P121 Characterisation of the miRNAome in Crohn’s Disease patients reveals transcriptomic changes associated with tissue composition and drug treatment

Abstract Background Gene regulation plays a major role in Crohn's Disease (CD) susceptibility and drug response. MicroRNA (miRNA)-mediated mechanisms tune gene expression at the post-transcriptional level. MiRNAs are 20-to-25-nt long non-coding RNAs that repress protein expression by direct inhibition or cleavage of the targets. Evaluating the miRNAome is emerging as a promising tool to clarify pathogenesis and identify promising biomarkers. We characterised the regulatory miRNAome across disease subtypes, gut tissue locations and infliximab treatment in CD. Methods We generated 120 transcriptomic profiles from terminal ileum and left colon biopsies from Hospital Universitario de la Princesa (Madrid). This cohort is composed of 10 patients with active endoscopy, 10 quiescent CD cases, and 10 controls (30 in total). In a balanced setting, half of the samples were incubated with infliximab for 18 hours at 37ºC and the rest with basal medium (Table 1). We carried out an exploratory analysis to gain insights into the determinants of miRNAome variability, including variance partition analyses and linear mixed modelling to detect significantly altered miRNAs. We also harnessed results from several publications including GSE16879 dataset to perform a comparative study with infliximab response signatures. Results Tissue location is the primary determinant of miRNA expression variability. The Variance Partition model identified 30 top miRNAs whose variability is maximised across regions. However, patient-specific effects also play a relevant role. We also identified 96 differentially expressed miRNAs between terminal ileum and left colon, with an overrepresentation of molecules involved in stress and developmental pathways (Fig.1). Among these, we find miR200s and miR429, that are associated with suppression of epithelial-mesenchymal transition and are up-regulated in colon more than in ileum. Down-regulation of miR196 is also shown in terminal ileum. Regarding the effects of infliximab treatment, we detected nine deregulated miRNAs which are associated with cellular response pathways. These include miR200s and miR192, which targets NOD2 and down-regulates NF-κB. Our results show an up-regulation of miR192 upon infliximab incubation (log2FC=2.37), which is associated with a reduction in TNFα expression and, consequently, with induced overexpression of miR192. We conclude that this miRNA is a potential target for therapy development. Conclusion In short, tissue location, and to a lesser extent disease subtype, are the main determinants of miRNAome in CD. A few treatment-responsive miRNAs like miR192 and miR200s are identified, in spite of detection challenges, emphasizing the need to improve our comprehension of the miRNA dynamics associated with drug response.

Discrimination of lipid composition and cellular localization in human liver tissues by stimulated Raman scattering microscopy.

The molecular mechanisms driving the progression from nonalcoholic fatty liver (NAFL) to fibrosing steatohepatitis (NASH) are insufficiently understood. Techniques enabling the characterization of different lipid species with both chemical and spatial information can provide valuable insights into their contributions to the disease progression. We extend the utility of stimulated Raman scattering (SRS) microscopy to characterize and quantify lipid species in liver tissue sections from patients with NAFL and NASH. We applied a dual-band hyperspectral SRS microscopy system for imaging tissue sections in both the C-H stretching and fingerprint regions. The same sections were imaged with polarization microscopy for detecting birefringent liquid crystals in the tissues. Our imaging and analysis pipeline provides accurate classification and quantification of free cholesterol, saturated cholesteryl esters (CEs), unsaturated CE, and triglycerides in liver tissue sections. The subcellular resolution enables investigations of the heterogeneous distribution of saturated CE, which has been under-examined in previous studies. We also discovered that the birefringent crystals, previously found to be associated with NASH development, are predominantly composed of saturated CE. Our method allows for a detailed characterization of lipid composition in human liver tissues and enables further investigation into the potential mechanism of NASH progression.

P049 The lipidome of creeping fat in Crohn’s disease points towards a harmful microenvironment

Abstract Background A pathognomonic observation in Crohn’s disease (CD) is the existence of mesenteric adipose tissue (MAT) wrapping around the inflamed and fibrotic intestine, so-called creeping fat. The etiology of this phenomenon is unclear, and both a protective and harmful role in disease pathogenesis and progression have been proposed. Given this conflicting evidence, we aimed to further unravel the role of creeping fat by extensive profiling of its lipidome. Methods We prospectively included 22 CD patients who underwent an ileocecal resection with ileocolonic anastomosis (Table 1). Perioperatively paired adipose tissue samples were collected from 3 locations: (1) creeping fat, (2) MAT close to the proximal unaffected ileum, and (3) MAT far from the diseased ileum, defined as near the base of the ileocecal artery (Fig. 1A). The lipidome of all 66 samples was profiled by quantitative mass spectrometry (targeted C18 LC-MS/MS and HILIC LC-MS/MS; Lipometrix core KU Leuven). This approach enables the detection of mediator lipids, and membrane and storage lipids, of which the latter group represents more complex chemical structures and thus a higher diversity in classification and number of species. Data analysis and paired Wilcoxon rank- sum tests were performed with Python and R 4.3.2. Results Across all CD adipose tissue samples, we detected 38 mediator lipid species, and 456 membrane and storage lipid species . When paired-wise comparing creeping fat with MAT far from the diseased ileum, we observed a significant decrease in 16 lipid mediators (p&amp;lt;0.05), namely arachidonic acid (AA), docosahexaenoic acid (DHA), 4 epoxy and 10 hydroxy mediator lipids; with hydroxy mediator X being the top downregulated in creeping fat (fold change (FC)=-2.11, p=0.0003) (Fig. 1B). While AA and DHA are (semi)-essential fatty acids and function as precursors, the other 8 decreased lipids in creeping fat are known to be anti-inflammatory, and 3 (incl. mediator X) are also anti-fibrotic. The lipid mediator profile of MAT close to the proximal unaffected ileum was similar to that of creeping fat, except for a pro-inflammatory lipid being increased in creeping fat (FC=2.30, p=0.004) (Fig. 1B). Lastly, when evaluating the membrane and storage lipid abundances both at species level (sum notation; fatty acyl composition) and at class level, no paired-wise differences among the three adipose tissue locations were found (p&amp;gt;0.05). Conclusion In this pilot lipidomics study, we did not observe any changes in storage or membrane lipids in creeping fat as compared to paired unaffected MAT. In contrast, a decrease in anti-inflammatory and anti-fibrotic lipid species along with an increase in a pro-inflammatory species points towards a harmful microenvironment within creeping fat.

Intraoperative segmental pancreatic occlusion and insulin assay combination optimizes the localization of lesions and confirmation of complete resection in pancreatic hypoglycemia patients

AimTo evaluate the efficacy of the application of intraoperative segmental pancreatic occlusion and insulin assay in surgical procedures for pancreatic hypoglycemia. MethodsWe retrospectively analyzed the clinical data of 11 pancreatic hypoglycemia cases treated in the China-Japan Friendship Hospital between September 2015 and August 2021. Intraoperative segmental pancreatic occlusion and insulin assay were used to enhance hypersecretory pancreatic tissues’ localization and to achieve a complete resection. Intraoperative testing of insulin levels (peripheral venous blood) was carried out at several time points starting from before the resection of hypersecretory tissues (base value) and at 1 minute, 5 minutes, 15 minutes, 30 minutes, and 60 minutes after resection. Additional testing every 30 minutes until the end of the operation was carried out when necessary. ResultsA total of 11 pancreatic hypoglycemia cases were included; 9 cases were insulinomas (all with single pancreatic lesions, with 4 located in the head, 1 in the body, and 4 in the tail), 1 MEN-1, and 1 nesidioblastosis. The insulin assay (30 minutes after the resection of hypersecretory tissues) enhanced the ability to locate target tissues and the accuracy of complete resection to 100%. As for intraoperative blood glucose monitoring, the accuracy 30 minutes after resection was as low as 36.6%. Postoperative levels of insulin and glucose were normal in all patients, with no recurrence of hypoglycemic symptoms during postoperative follow-up visits (9 to 72 months). ConclusionIntraoperative segmental pancreatic occlusion and insulin assay in pancreatic hypoglycemia is a simple, accurate, and fast approach that enhances the localization and complete resection of hypersecretory tissues. Such a combination is highly significant in challenging cases of hypoglycemia.

Cloning, distribution, and effects of growth regulation of MC3R and MC4R in red crucian carp (Carassius auratus red var.).

Melanocortin-3 and -4 receptors (MC3R and MC4R), G protein-coupled receptors, play vital roles in the regulation of energy homeostasis. To understand the functions of mc3r and mc4r in the energy homeostasis of red crucian carp (Carassius auratus red var., RCC), we cloned mc3r and mc4r, analyzed the tissue expression and localization of the genes, and investigated the effects of knockout of mc3r (mc3r +/-) and mc4r (mc4r +/-) in RCC. The full-length cDNAs of RCC mc3r and mc4r were 1459 base pairs (bp) and 1894 bp, respectively. qRT-PCR indicated that mc3r and mc4r were profusely expressed in the brain, but lower expressed in the periphery tissues. ISH revealed that mc3r and mc4r were located in NPP, NPO, NAPv, NSC, NAT, NRL, NLTl, and NLTp of the brain, suggesting that mc3r and mc4r might regulate many physiological and behavioral aspects in RCC. To further verify the roles of mc3r and mc4r in energy homeostasis, the mc3r+/- and mc4r+/- fish were obtained by the CRISPR/Cas9 system. The average body weights, total lengths, body depths, and food intake of mc4r+/- fish were significantly higher than those of mc3r+/- and the normal wild-type (WT) fish, but there was no difference between the mc3r+/- and WT fish, indicating that the RCC phenotype and food intake were mainly influenced by mc4r but not mc3r. Interestingly, mc4r+/- fish displayed more visceral fat mass than mc3r+/- and WT fish, and mc3r+/- fish also exhibited slightly more visceral fat mass compared to WT. RNA-seq of the liver and muscle revealed that a large number of differentially expressed genes (DEGs) differed in WT vs. mc3r+/-, WT vs. mc4r+/-, and mc3r+/- vs. mc4r+/-, mainly related to lipid, glucose, and energy metabolism. The KEGG enrichment analysis revealed that DEGs were mainly enriched in pathways such as steroid biosynthesis, fatty acid metabolism, fatty acid biosynthesis, glycolysis/gluconeogenesis, wnt signaling pathway, PPAR signaling pathway, and MAPK signaling pathway, thereby affecting lipid accumulation and growth. In conclusion, these results will assist in the further investigation of the molecular mechanisms in which MC3R and MC4R were involved in the regulation of energy homeostasis in fish.

Identification and Analysis of SND1 as an Oncogene and Prognostic Biomarker for Lung Adenocarcinoma

Transcription factor (TF) can bind specific sequences that either promotes or represses the transcription of target genes, and exerts important effects on tumorigenesis, migration, invasion. Staphylococcal nuclease-containing structural domain 1 (SND1), which is a transcriptional co-activator, is considered as a promising target for tumor therapy. However, its role in lung adenocarcinoma (LUAD) remains unclear. This study aims to explore the role of SND1 in LUAD. Data from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), Clinical Proteomic Tumor Analysis Consortium (CPTAC), and Human Protein Atlas (HPA) database was obtained to explore the association between SND1 and the prognosis, as well as the immune cell infiltration, and subcellular localization in LUAD tissues. Furthermore, the functional role of SND1 in LUAD was verified in vitro. EdU assay, CCK-8 assay, flow cytometry, scratch assay, Transwell assay and Western blot were performed. SND1 was found to be upregulated and high expression of SND1 is correlated with poor prognosis of LUAD patients. In addition, SND1 was predominantly present in the cytoplasm of LUAD cells. Enrichment analysis showed that SND1 was closely associated with the cell cycle, as well as DNA replication, and chromosome segregation. Immune infiltration analysis showed that SND1 was closely associated with various immune cell populations, including T cells, B cells, cytotoxic cells and dendritic cells. In vitro studies demonstrated that silencing of SND1 inhibited cell proliferation, invasion and migration of LUAD cells. Besides, cell cycle was blocked at G1 phase by down-regulating SND1. SND1 might be an important prognostic biomarker of LUAD and may promote LUAD cells proliferation and migration.

Non-purulent myositis caused by direct invasion of skeletal muscle tissue by Leptospira in a hamster model.

Myalgia is a common symptom of Leptospira infection in humans. Autopsies have reported that muscle tissue shows degeneration and necrosis of the myofibers and infiltration of inflammatory cells composed mainly of macrophages and lymphocytes. It remains unclear whether Leptospira directly infects the muscle and how the infiltrating inflammatory cells are involved in muscle fiber destruction. This study evaluated the relationship between histopathological changes and leptospiral localization in the muscle tissue of a hamster model. The influence of macrophages in skeletal muscle injury was also investigated, using selective depletion of macrophages by administration of liposomal clodronate. Hamsters infected subcutaneously with Leptospira interrogans serovar Manilae strain UP-MMC-SM showed myositis of the thighs adjacent to the inoculated area beginning at 6 days post-infection. The myositis was non-purulent and showed sporadic degeneration and necrosis of muscle fibers. The degeneration of myofibers was accompanied by aggregations of macrophages. Immunofluorescence staining revealed leptospires surrounding the damaged muscle fibers. Subcutaneous injection of formalin-killed Leptospira or intraperitoneal injection of live Leptospira caused no myositis in hamster thighs. Liposomal clodronate treatment in infected hamsters reduced macrophage infiltration in muscle tissue without impacting bacterial clearance. Muscle necrosis was still observed in the infected hamsters treated with liposomal clodronate, and there was no significant change in serum creatine kinase levels compared to those in animals treated with liposomes alone. Our findings suggest that leptospiral invasion of muscle tissue from an inoculation site leads to the destruction of muscle fibers and causes non-purulent myositis, whereas the infiltrating macrophages contribute less to muscle destruction.

Breast calcifications. Literature review

Breast cancer continues to maintain a leading position in the structure of the incidence of all oncological diseases among women. Early diagnosis of the disease at the stage of assessing indirect signs makes it possible in the future to maintain a good quality of life for patients by forming proper dynamic monitoring and performing functionally sparing surgical treatment. Interest in the early detection of benign and malignant pathology of the mammary gland is growing every year. A multidisciplinary approach to the study of the formation of breast calcifications makes it possible to look at the problem in a multifaceted way, and ways to solve problems in the process of diagnosis underlie the biochemical understanding of the mechanisms of formation, which is no less important when evaluating mammographic images. Practically up to 50 % of non-palpable breast tumors in practice are detected by signs of existing calcifications in the gland tissue. At present, the problem of early detection of malignant tumors of the breast by radiographic signs of these calcifications, their shape and nature of location in the gland tissue and their further pathomorphological confirmation is being actively studied.We have analyzed the literature data of foreign and domestic authors over the past 20 years. The problem was considered from the mechanism of formation and morphological features of calcifications in the breast tissue to their radiographic assessment on mammographic images. According to the results of the studied literature, the biochemical features of the formations were taken as the basis for understanding the nature of calcifications in the breast tissue. On a biochemical level, calcifications are usually classified into two main types: type I, consisting of calcium oxalate, and type II, consisting of hydroxyapatite. The classification is based on chemical composition and mammographic characteristics, including morphology, distribution and density. Speaking about calcium oxalate, the benign nature of education is most often implied, which cannot be said about hydroxyapatite. Mammography is the main method for diagnosing these formations, taking into account the characteristics of the characteristic radiological signs, which are also detailed in this article.Understanding the molecular and structural development of calcifications may aid in the detection and treatment of breast lesions. Identification of these structures in a patient diagnosed with breast cancer refers to the early diagnosis of the disease, where, with timely prescribed therapy, we get high chances of good long-term results of relapse-free survival.

Pathology and Monkeypox virus Localization in Tissues From Immunocompromised Patients With Severe or Fatal Mpox.

Pathology and Monkeypox virus (MPXV) tissue tropism in severe and fatal human mpox is not thoroughly described but can help elucidate the disease pathogenesis and the role of coinfections in immunocompromised patients. We analyzed biopsy and autopsy tissues from 22 patients with severe or fatal outcomes to characterize pathology and viral antigen and DNA distribution in tissues by immunohistochemistry and in situ hybridization. Tissue-based testing for coinfections was also performed. Mucocutaneous lesions showed necrotizing and proliferative epithelial changes. Deceased patients with autopsy tissues evaluated had digestive tract lesions, and half had systemic tissue necrosis with thrombotic vasculopathy in lymphoid tissues, lung, or other solid organs. Half also had bronchopneumonia, and one-third had acute lung injury. All cases had MPXV antigen and DNA detected in tissues. Coinfections were identified in 5 of 16 (31%) biopsy and 4 of 6 (67%) autopsy cases. Severe mpox in immunocompromised patients is characterized by extensive viral infection of tissues and viremic dissemination that can progress despite available therapeutics. Digestive tract and lung involvement are common and associated with prominent histopathological and clinical manifestations. Coinfections may complicate mpox diagnosis and treatment. Significant viral DNA (likely correlating to infectious virus) in tissues necessitates enhanced biosafety measures in healthcare and autopsy settings.

Unsupervised unpaired multiple fusion adaptation aided with self-attention generative adversarial network for scar tissues segmentation framework

Late gadolinium enhancement (LGE) is a specialized imaging technique used in cardiovascular magnetic resonance (CMR) imaging to detect and characterize areas of scar tissue or fibrosis within the heart muscle. After a heart attack, the affected region of the heart muscle becomes scarred due to insufficient blood supply. LGE CMR enhances the infarcted myocardium to appear with distinctive brightness compared to healthy tissues. It is primarily used to assess various heart conditions and can identify the extent and location of scar tissue, helping in risk stratification and guiding treatment decisions. Automated myocardium segmentation in LGE CMR poses challenges due to intensity heterogeneity caused by the accumulation of contrast agents in infarcted areas. Furthermore, LGE CMR images with gold standard labels are particularly scarce compared to other sequences, making the task even more demanding. Besides, the heart’s motion involves complex non-rigid deformations, including regional wall thickening, circumferential, and longitudinal ventricular shortening, making it challenging to model. This work presents an unsupervised unpaired domain adaptive scar tissue segmentation framework aided with the self-attention generative adversarial network. Besides, we proposed a novel segmentation architecture based on a parallel transformer for scar segmentation and compared the results with and without style transfer GAN and unpaired style self-supervised attention GAN. Besides, to model longitudinal changes in scare, we used eight days’ time points as a source and 1 month and 12 as a target and applied our proposed attention-generative network to transfer style from one-time point to another time. Experiments on four different LGE MICCAI datasets from 2019 to 2023 achieved significantly better performance than state-of-the-art segmentation methods.

Real-Time Dynamics of Water Transport in the Roots of Intact Maize Plants in Response to Water Stress: The Role of Aquaporins and the Contribution of Different Water Transport Pathways.

Using an original methodological and technical approach, we studied the real-time dynamics of radial water transfer in roots and transpiration rate in intact maize plants in response to water stress. It was shown that the response of maize plants to water stress, induced by 10% PEG 6000, was accompanied by changes in the intensity and redistribution of water transfer along different pathways of radial water transport in the roots. It was shown that during the first minutes of water stress impact, the intensity of transcellular and symplastic water transport in the roots decreased with a parallel short-term increase in the transpiration rate in leaves and, presumably, in apoplastic transport in roots. Further, after a decrease in transpiration rate, the intensity of transcellular and symplastic water transport was restored to approximately the initial values and was accompanied by parallel upregulation of some PIP aquaporin genes in roots and leaves, changes in aquaporin localization in root tissues, and changes in xylem sap pH. Under water stress conditions, cell-to-cell water transport in roots becomes dominant, and aquaporins contribute to the simultaneous regulation of water transport in roots and shoots under water stress.