Analysis Of Histological Sections Research Articles

Effects of chronic heat stress on spleen structure, apoptosis and immune response in Siberian Sturgeon (Acipenser baerii)

Chronic heat stress induced by global warming has important implications for fish survival, and the response of important immune organs, such as the spleen, to chronic heat stress remains to be investigated. In this study, one control group (20 ℃) and two heat stress groups (24 ℃ and 28 ℃) were set up to investigate the effects of heat stress on the spleen of Siberian Sturgeon (Acipenser baerii). As per the histological section analysis, heat stress caused vacuolization, reticulocyte hyperplasia, and macrophage infiltration in the spleen, with hyperplasia being more pronounced in the 28 ℃ group. The flow cytometry results showed that the spleen’s apoptosis level significantly increased at 28 ℃ (p < 0.05). Hematological studies revealed a significant increase in the number of red blood cells in the 28 ℃ group (p < 0.05). Meanwhile, the mRNA expression of immune-related factors (tgf-β, il-1β, and il-8) was significantly higher in the 28 ℃ heat stress group compared with the control group. This study reveals the effects of heat stress on the A. baerii spleen and contributes to the understanding of coping strategies of cold-water fish to chronic heat stress.

Origin, structure, and composition of the spider major ampullate silk fiber revealed by genomics, proteomics, and single-cell and spatial transcriptomics.

Spiders produce nature's toughest fiber using renewable components at ambient temperatures and with water as solvent, making it highly interesting to replicate for the materials industry. Despite this, much remains to be understood about the bioprocessing and composition of spider silk fibers. Here, we identify 18 proteins that make up the spiders' strongest silk type, the major ampullate fiber. Single-cell RNA sequencing and spatial transcriptomics revealed that the secretory epithelium of the gland harbors six cell types. These cell types are confined to three distinct glandular zones that produce specific combinations of silk proteins. Image analysis of histological sections showed that the secretions from the three zones do not mix, and proteomics analysis revealed that these secretions form layers in the final fiber. Using a multi-omics approach, we provide substantial advancements in the understanding of the structure and function of the major ampullate silk gland as well as of the architecture and composition of the fiber it produces.

<i>In situ</i> and <i>in silico</i> modeling of the hematopoiesis-inducing effect of chelidonic acid

The current trend in regenerative medicine, in the context of an aging population, is the search for new ways and means to optimize tissue bioengineering. One of the convenient models for in situ studying bone marrow regeneration is the subcutaneous ectopic osteogenesis test on scaffolds that imitate the architecture of bone tissue. Chelidonic acid (CA), a small molecule, is capable of participating in various cellular processes and metabolic pathways, and it can activate the osteogenic differentiation of mesenchymal stem cells. However, the molecular mechanisms behind the regulatory effects of CA remain unknown. The aim of this study was to investigate the modulatory effect of CA on the in situ formation of hematopoietic foci, as well as to predict target genes and intracellular signalling pathways involved in the hematopoietic activity of CA. An aqueous solution of CA, isolated from an extract of the Saussurea controversa plant. Course (daily for 35 days) oral administration of CA. Ectopic osteogenesis testing in Balb/c mice. Morphometric analysis of histological sections after 45 days and in silico modelling of gene expression with statistical analysis. CA, when administered orally in a low dose (10 mg/kg), threefold increases the normalized area of bone marrow in the composition of bone tissue plates grown in situ in a test of ectopic subcutaneous osteogenesis in mice. This effect is associated essentially (a probability of CA activity Pa 0.5 and a probability of inactivity Pi 0.5) with enhanced expression of 358 hematopoiesis-related genes, as predicted by in silico analysis. The top list with the highest Pa value included 10 target genes, such as GATA1, CITED2, SFRP1, EP300, LGALS9, VNN1, IL10RB, RARA, CD83, and HMOX1. CA has a significant ability to enhance the reparative remodelling of hematopoietic tissue in situ. The next phase of research will be to test actual target genes and signalling pathways that mediate the regulatory effect of HC on hematopoiesis both in vitro and in vivo, as well as in clinical settings.

Long-term residence and efficacy of adenovirus-mimetic nanoparticles in renal target tissue

A major shortcoming in the treatment of mesangial cell-associated diseases such as IgA nephropathy, diabetic nephropathy, or lupus nephritis, which frequently progress to end-stage renal disease, is poor drug availability in the glomerular mesangium. Drug delivery via active targeting of nanoparticles, using ligands attached to the particle surface for target cell recognition to increase the biodistribution to the mesangium, is a promising strategy to overcome this hurdle. However, although several glomerular tissue targeting approaches have been described, so far no study has demonstrated the particles’ ability to deliver sufficient drug amounts combined with an appropriate nanoparticle target retention time to trigger relevant biological effects in the mesangium. In our study, we encapsulated erastin, a ferroptosis-inducing model compound, into adenovirus-mimetic, mesangial cell-targeting nanoparticles, enabling the direct visualisation of biological effects through ferroptosis-dependent histological changes. By intravital microscopy and analysis of histological sections, we were not only able to localise the injected particles over 10 days within the target cells but also to demonstrate biological activity in the renal glomeruli. In conclusion, we have characterised adenovirus-mimetic nanoparticles as a highly suitable drug delivery platform for the treatment of mesangial cell-associated diseases and additionally provided the basis for a potential renal disease model.

Advancing drug development by leveraging microdissection for proteogenomic analysis of histological sections.

e14660 Background: Combining laser capture microdissection (LCM) with proteogenomics in cancer research offers a targeted way to way to explore the intricate interactions between the tumor microenvironment and tumor cells proper. For immuno-oncology (IO) research, this is especially important, since improvements in the predictability of patient response to IO-based drugs is greatly needed. An improved understanding of the spatial relationships involving tumor cells, stromal cells, blood supply and immune cell interactions may help to inform drug development and improve the indicators of success for IO-based drug regimens. Methods: From fresh frozen tissue, LCM is used to isolate and obtain distinct histological cell types. Nucleic acids and proteins are extracted from captured cells for in-depth multi-modality molecular profiling assays. This includes multiple genomic modalities and a customized solution-based mass spectrometry proteomics approach for samples or specimens with a limited amount of tissue. Results: Optimizing the analysis of minute tissue quantities from LCM captured cells is challenging. Following the isolation of nucleic acids, DNA sequencing can be performed for the discovery and analysis of actionable mutations, copy number variation, and methylation profiles. RNA-seq may be performed for gene expression. Regarding genomic-based studies, commercial kits are often utilized for these steps due to their general availability, rapid innovation, and the robust performance of these products. However, this kit-based adaptive approach is not true for proteomics. There remains a need for highly sensitive proteomic methods targeting small-sized samples. Our novel proteogenomics approach is focused around a customized enhanced liquid chromatography mass spectrometry (LC-MS) analysis of micro-scale and/or nano-scale tissue sections. This is achieved with a silver-stained one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D-SDS-PAGE) approach developed for LC-MS analysis of fresh-frozen tissue specimens obtained via LCM. This includes an in-gel digestion method adjusted and specifically designed to maximize the proteome coverage from amount-limited LCM samples to better facilitate in-depth molecular profiling. Conclusions: A proteogenomic approach is reported that is utilizing amount limited tissue quantities from microdissected fresh frozen tissue. The methodology may also be applicable to other types of specimens having limited nucleic acids, protein quantity, and/or sample volume.

The fix is not yet in: recommendation for fixation of lungs within physiological/pathophysiological volume range in preclinical pulmonary structure-function studies.

Quantitative characterization of lung structures by morphometrical or stereological analysis of histological sections is a powerful means of elucidating pulmonary structure-function relations. The overwhelming majority of studies, however, fix lungs for histology at pressures outside the physiological/pathophysiological respiratory volume range. Thus, valuable information is being lost. In this perspective article, we argue that investigators performing pulmonary histological studies should consider whether the aims of their studies would benefit from fixation at functional transpulmonary pressures, particularly those of end-inspiration and end-expiration. We survey the pressures at which lungs are typically fixed in preclinical structure-function studies, provide examples of conditions that would benefit from histological evaluation at functional lung volumes, summarize available fixation methods, discuss alternative imaging modalities, and discuss challenges to implementing the suggested approach and means of addressing those challenges. We aim to persuade investigators that modifying or complementing the traditional histological approach by fixing lungs at minimal and maximal functional volumes could enable new understanding of pulmonary structure-function relations.

INDICATORS OF MUC 5AC AND MUC 1 MUCOPOLYSACCHARIDE EXPRESSION IN THE NASAL MUCOSA OF PATIENTS WITH POSTNASAL DRIP SYNDROME AND NASAL SEPTUM DEVIATION

Introduction. Determining the cause of postnasal drip syndrome can be quite challenging. Nasal septum deformation has been recognized as one possible factor. Understanding the fundamental processes of postnasal drip syndrome is crucial for developing targeted therapeutic strategies for treating this pathology. In our study, we examined the clinical and histological features of postnasal drip syndrome in individuals with nasal septum deformation and investigated the expression of mucopolysaccharides, specifically the indicators of MUC 5AC and MUC 1.
 The aim. This study aims to investigate the clinical and histological aspects of postnasal drip syndrome (PNDS) in patients with nasal septum deformation. Specifically, the study aims to explore the expression levels of MUC5AC and MUC1 in biopsies of the nasal turbinate mucosa.
 Materials and methods. A total of 29 samples of nasal mucosa from the lower nasal turbinates were collected. These samples were divided into two groups: patients with nasal septum deviation with and without postnasal drip syndrome. Levels of MUC5AC and MUC1 were determined using Western blot analysis.
 Results. Upon analysis of histological sections, we identified a significant increase in tissue metaplasia and lymphoid infiltration in the nasal mucosa of patients with postnasal drip syndrome compared to the control group. The levels of mucopolysaccharides, MUC5AC and MUC1, were higher in the nasal mucosa of the research group compared to the control group.
 Conclusions. The obtained data suggest that anatomical changes in the nasal cavity may play a role in the development of postnasal drip syndrome through alterations in mucin secretion.

Abstract 730: Leveraging microdissection for proteogenomic analysis of histological sections to advance drug development

Abstract Background: Combining proteogenomics with laser capture microdissection (LCM) in cancer research offers a targeted way to explore the intricate interactions between tumor cells and the tumor microenvironment. This is especially important for immuno-oncology (IO) research where improvements in the predictability of patient response to IO-based drugs is sorely needed. An improved understanding of the spatial relationships involving tumor cells proper, stromal cells, blood supply and immune cell interactions may help to inform drug development and improve the indicators of success for IO-based drug regimens. Methods: LCM is used to isolate and obtain distinct histological cell types from fresh frozen tissue. Once cells have been captured, nucleic acids and proteins are extracted for in-depth multi-modality molecular profiling assays involving multiple genomic modalities and a customized solution-based mass spectrometry proteomics approach for samples or specimens with a limited amount of tissue. Results: Optimizing analysis of minute tissue quantities from LCM captured cells is challenging. Following the isolation of nucleic acids, RNA-seq may be performed for gene expression and DNA sequencing performed for the discovery and analysis of actionable mutations, copy number variation, and methylation profiles. Regarding genomic-based studies, commercial kits are often utilized for these steps due to their general availability, rapid innovation, and the robust performance of these products. However, this kit-based adaptive approach is not true for proteomics. There remains a need for highly sensitive proteomic methods targeting small-sized samples. A significant part of our proteogenomics approach is focused around an enhanced liquid chromatography mass spectrometry (LC-MS) analysis of micro-scale and/or nano-scale tissue sections. This is achieved with a silver-stained one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D-SDS-PAGE) approach developed for LC-MS analysis of fresh-frozen tissue specimens obtained via LCM. This includes an in-gel digestion method adjusted and specifically designed to maximize the proteome coverage from amount-limited LCM samples to better facilitate in-depth molecular profiling. Conclusions: Reported here is a proteogenomic approach that is leveraging from microdissected fresh frozen tissue. The methodology may also be applicable to other types of specimens having limited nucleic acids, protein quantity, and/or sample volume. Citation Format: Ik Jae Shin, Michael Tangrea, Michael R. Emmert-Buck, Donald J. Johann. Leveraging microdissection for proteogenomic analysis of histological sections to advance drug development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 730.

Huaier improves the efficacy of anti-PD-L1 Ab in the treatment of hepatocellular carcinoma by regulating tumor immune microenvironment

BackgroundCombination therapy is an effective method for augmenting the efficacy of immune checkpoint inhibitors (ICIs). Huaier is a commonly used Chinese patent medicine with substantial antitumor effects. The combination of Huaier and ICIs may increase the efficacy of ICIs against hepatocellular carcinoma (HCC). MethodsThe major components of Huaier were detected by high-performance liquid chromatography-mass spectrometry. The optimal antitumor dose of Huaier was investigated in H22-bearing mice. Next, Huaier was combined with anti-CD8α antibody (Ab) or anti-PD-L1 Ab to observe the antitumor effect. The safety of these combination drugs was evaluated through blood biochemical tests and hematoxylin and eosin staining of histological sections. RT-qPCR, immunohistochemistry, flow cytometry, and transcriptome sequencing were performed to investigate the potential action mechanism of anti-PD-L1 Ab combined with Huaier against HCC. ResultsHPLC-MS/MS identified 333 components of Huaier, including carboxylic acids and derivatives, thienothiophenes, phenols, flavonoids and so on. Huaier exhibited significant antitumor effects, with the strongest effect noted at a dose of 4 g/kg. Huaier boosted CD8+ T cells infiltration into the tumor. Next, CD8+ T cells were depleted by with anti-CD8α Ab, and the antitumor effect of Huaier was suppressed. Flow cytometry results revealed that CD8+ T cells were reduced in the Huaier+anti-CD8α Ab group, with the antitumor effect of this group being inhibited. This indicated that CD8+ T cells were key players in the antitumor activity of Huaier. Meanwhile, Huaier inhibited microvessel density (MVD), downregulated vascular endothelial growth factor A (VEGFA), and upregulated PD-L1 in tumor tissues. Finally, Huaier combined with anti-PD-L1 Ab exhibited a greater antitumor effect in the H22-bearing mice. And the results of liver and kidney function tests and histological section analysis unveiled that the safety of these drugs was excellent. According to the transcriptome sequencing results, Huaier combined with anti-PD-L1 Ab possibly exerted anti-HCC effects through immunomodulation, immune response, and so on. ConclusionsHuaier exhibited a significant antitumor effect. It promoted CD8+ T cells infiltration, upregulated PD-L1 expression, downregulated VEGFA expression, and inhibited MVD, thereby playing a significant antitumor immunoregulatory effect. The combination of Huaier and anti-PD-L1 Ab has significant antitumor effects, and this regimen has good safety. Therefore, Huaier combined with anti-PD-L1 Ab is a promising therapeutic approach against HCC.

Effect of Metformin on Adipose Tissue Degeneration During Obesity

Obesity is associated with an increased inflammation involved in the development of atherosclerosis and insulin resistance. Macrophages are key in the genesis of these processes. Obesity induces macrophage accumulation in adipose tissue, they produce many of the inflammatory molecules secreted by adipose tissue like TNF-α, interleukins (IL-1β, IL-6, IL-8), as well as acute phase proteins such as haptoglobin (Hp), PAI-1 and C-reactive protein. The presence of these molecules in obesity relates this pathology to various inflammation degrees.The analysis of histological sections obtained from rabbit adipose tissue biopsy samples and processed by the hematoxylin-eosin and immunohistochemistry, showed the effect of metformin, a molecule which acts on the insulin metabolism, by the degeneration of this tissue during obesity. Metformin and its effects on metabolism are such that decreases the inflammation process suffered by adipose tissue, which is reflected by morphological changes in the distribution and migration of cells in the histological structure.

Pharmacological Effects of Grape Leaf Extract Reduce Eimeriosis-Induced Inflammation, Oxidative Status Change, and Goblet Cell Response in the Jejunum of Mice.

Coccidiosis is a parasitic infection threatening poultry products globally. Parasite resistance to drugs is one of the barriers to Eimeria control. Natural products are one of the sources of compounds that prevent parasite infections. The current study was, therefore, conducted to evaluate the effect of Vitis vinifera leaf extract on anti-inflammatory response, oxidative status, and goblet cell response against Eimeria papillate infection in mice. Methanol was used as a solvent for phytochemicals. The mice were divided into six groups: The first group was the control. The second group was uninfected and treated with 200 mg/kg of extract to test toxicity, and the third, fourth, fifth, and sixth groups of mice received 1 × 103 sporulated E. papillate oocysts. The third group received no treatment. The fourth and fifth groups were treated daily with 100 and 200 mg/kg of V. vinifera leaf extract, respectively, while the sixth group received 25 mg/kg of toltrazuril daily via gavage. On day 5 p.i., the animals were sacrificed, and jejunum samples were prepared for analyses of histological sections and oxidative stress. The phytochemical analysis using GC-MS of the extract showed the presence of 12 biologically active compounds. The most effective dose was 200 mg/kg, which significantly decreased the number of parasitic stages in the jejunal sections of the mice. The findings demonstrate that E. papillate infection in mice results in significant histopathological changes in the jejunum, including inflammation, epithelial vacuolation, villi loss, and a decrease in goblet cell density. When infected mice received treatment, the histological injury score within the infected jejunum tissue decreased by 63%, and the goblet cell quantity dramatically increased, approaching the control values. Finally, the extract ameliorated the changes in glutathione and malondialdehyde due to E. papillate infection. The extract was proven to have anti-inflammatory properties and reduce the number of oocysts. Overall, the findings show that V. vinifera leaf extract has significant anticoccidial effects in vivo.

Probiotic Bacteria from Human Milk Can Alleviate Oral Bovine Casein Sensitization in Juvenile Wistar Rats.

This study aims to see if probiotic bacteria from human milk could ameliorate oral cow's milk sensitization. The probiotic potential of the SL42 strain isolated from the milk of a healthy young mother was first determined. Rats were then randomly gavaged with cow's milk casein without an adjuvant or assigned to the control group. Each group was further subdivided into three groups, with each receiving only Limosilactobacillus reuteri DSM 17938, SL42, or a phosphate-buffered saline solution. Body weight, temperature, eosinophils, serum milk casein-specific IgE (CAS-IgE), histamine, and serum S100A8/A9 and inflammatory cytokine concentrations were measured. The animals were sacrificed after 59 days; histological sections were prepared, and the spleen or thymus weights, as well as the diversity of the gut microbiota, were measured. On days 1 and 59, SL42 abridged systemic allergic responses to casein by dropping histamine levels (25.7%), CAS-specific IgE levels (53.6%), eosinophil numbers (17%), S100A8/9 (18.7%), and cytokine concentrations (25.4-48.5%). Analyses of histological sections of the jejunum confirmed the protective effect of probiotic bacteria in the CAS-challenged groups. Lactic acid bacteria and Clostridia species were also increased in all probiotic-treated groups. These findings suggest that probiotics derived from human milk could be used to alleviate cow's milk casein allergy.

Inactivation of Vibrio parahaemolyticus and retardation of quality loss in oyster (Crassostrea gigas) by ultrasound processing during storage

The health problems caused by foodborne pathogens of raw oysters have been widely concerned. Traditional heating methods tend to lead the loss of the original nutrients and flavors, in this study, the nonthermal ultrasound technology was applied to inactivate Vibrio parahaemolyticus on raw oysters, and the retardation effects on microbial growth and quality loss of oysters stored at 4 ℃ after ultrasonic treatment were also investigated. After treated by 7.5 W/mL ultrasound for 12.5 min, the Vibrio parahaemolyticus in oysters was reduced by 3.13 log CFU/g. By measuring total aerobic bacteria and total volatile base nitrogen, the growth trend after ultrasonic treatment was delayed compared with heat treatment, and the shelf life of oysters was prolonged. At the same time, ultrasonic treatment delayed the changes of color difference and lipid oxidation of oysters during cold storage. Texture analysis showed that ultrasonic treatment helped maintain the good textural structure of oysters. Histological section analysis also demonstrated that muscle fibers were still tightly packed after ultrasonic treatment. Low-field nuclear magnetic resonance (LF-NMR) illustrated that the water in the oysters was well maintained after ultrasonic treatment. In addition, gas chromatograph – ion mobility spectrometer (GC-IMS) showed that ultrasound treatment could better preserve the flavor of oysters during cold storage. Therefore, it is believed that ultrasound can inactivate foodborne pathogens of raw oysters and keep its freshness and original taste better during storage.

Pharmacological Blockade of TGF-Beta Reduces Renal Interstitial Fibrosis in a Chronic Ischemia–Reperfusion Animal Model

The targeting of transforming growth factor β (TGF-β) has been shown to reduce complications related to ischemia-reperfusion injury (IRI) post-surgically. Pirfenidone (PFD) specifically inhibits TGF-β expression and has been demonstrated to provide protection from IRI in short-term allograft models, though not yet in long-term models. A chronic unilateral IRI model was established using male Wistar rats. The animals were divided into two groups: one with IRI and a pre-treatment of PFD (0.5 mg/kg) followed by 0.5 mg/kg/day of orally administered PFD for 30 days, and a control group without PFD treatment. A sham group was also included. Kidneys and blood samples were collected after 30 days, and the renal function was evaluated by measuring the serum creatinine and KIM-1 levels. RT-PCR was used to analyze fibrosis-related genes, and Luminex to quantify the pro-inflammatory serum IL-18 cytokine. Renal section staining and histological analysis were used to detect collagen deposits. Comparison within the groups showed an increase in serum creatinine and KIM-1 expression after IRI in the control group, while PFD reduced COLL1A1 and TGF-β expression and demonstrated a reduction in fibrosis through histological stains. The treatment group also showed a reduction in IL-18. Our results suggest that PFD exerts protective effects on chronic renal IRI, reducing fibrosis development and inflammation. This study provides new insights into the treatment and management of chronic renal function loss after IRI.

Biological mechanisms of infection resistance in tissue engineered blood vessels compared to synthetic expanded polytetrafluoroethylene grafts

ObjectiveSynthetic expanded polytetrafluoroethylene (ePTFE) grafts are known to be susceptible to bacterial infection. Results from preclinical and clinical studies of bioengineered human acellular vessels (HAVs) have shown relatively low rates of infection. This study evaluates the interactions of human neutrophils and bacteria with ePTFE and HAV vascular conduits to determine whether there is a correlation between neutrophil-conduit interactions and observed differences of their infectivity in vivo. MethodsA phase III comparative clinical study between investigational HAVs (n = 177) and commercial ePTFE grafts (n = 178) used for hemodialysis access (ClinicalTrials.gov Identifier: NCT02644941) was evaluated for conduit infection rates followed by histological analyses of HAV and ePTFE tissue explants. The clinical histopathology of HAV and ePTFE conduits reported to be infected was compared with immunohistochemistry of explanted materials from a preclinical model of bacterial contamination. Mechanistic in vitro studies were then conducted using isolated human neutrophils seeded directly onto HAV and ePTFE materials to analyze neutrophil viability, morphology, and function. ResultsClinical trial results showed that the HAV had a significantly lower (0.93%; P = .0413) infection rate than that of ePTFE (4.54%). Histological analysis of sections from infected grafts explanted approximately 1 year after implantation revealed gram-positive bacteria near cannulation sites. Immunohistochemistry of HAV and ePTFE implanted in a well-controlled rodent infection model suggested that the ePTFE matrix permitted bacterial infiltration and colonization but may be inaccessible to neutrophils. In the same model, the HAV showed host recellularization and lacked detectable bacteria at the 2-week explant. In vitro results demonstrated that the viability of human neutrophils decreased significantly upon exposure to ePTFE, which was associated with neutrophil elastase release in the absence of bacteria. In contrast, neutrophils exposed to the HAV material retained high viability and native morphology. Cocultures of neutrophils and Staphylococcus aureus on the conduit materials demonstrated that neutrophils were more effective at ensnaring and degrading bacteria on the HAV than on ePTFE. ConclusionsThe HAV material seems to demonstrate a resistance to bacterial infection. This infection resistance is likely due to the HAV's native-like material composition, which may be more biocompatible with host neutrophils than synthetic vascular graft material.

РОЛЬ ШВАННОВСКИХ КЛЕТОК В ПАТОЛОГИИ ЧЕРВЕОБРАЗНОГО ОТРОСТКА У ДЕТЕЙ

Objective: To compare the immunohistochemical (IHC) characteristics of appendices removed from children with acute appendicitis and chronic pain syndrome (CP) in the right lower quadrant of the abdomen (RLQA), as well as to define clinical and morphological correlations. Methods: The structure of fifty-one appendices of children aged 5-14 years who underwent appendectomy for chronic appendicitis/CP in the RLQA (24 patients, Group 1) and acute appendicitis (27 patients, Group 2) was assessed. Image analysis of histological sections stained with hematoxylin-eosin and immunohistochemically for S100 protein and proliferating cell nuclear antigen (PCNA) was performed. Results: Image analysis showed the presence of a significantly higher volume density (VD) and numerical density (ND) (p0.05) of immunoreactive cells when stained for S100 protein, as well as the ND of intramural ganglia (p0.05) against the background of minimal histological changes in the wall of the appendix in patients of Group 1 compared to Group 2. On the contrary, the ND of PCNA-immunoreactive cells was significantly higher in patients of Group 2 against the background of pronounced inflammatory changes in the appendix (p0.01). In the biopsy samples of the Group 1 patients, a significantly higher ND of the lymphoid nodule germinal centers was also noted (p0.05), while the ND of tingible body macrophages was higher in Group 2 patients (p0.001). These results demonstrate differences in neuroimmune parameters in the appendices in the two described nosologic groups and indicate the possible role of the appendiceal neural component as a prerequisite for developing CP in the RLQA in children. Keywords: Appendix, Schwann cell, S100 protein, PCNA, appendicitis, chronic abdominal syndrome.

Resorbable Nanomatrices from Microbial Polyhydroxyalkanoates: Design Strategy and Characterization.

From a series of biodegradable natural polymers of polyhydroxyalkanoates (PHAs)-poly-3-hydroxybutyrate (P(3HB) and copolymers containing, in addition to 3HB monomers, monomers of 3-hydroxyvalerate (3HV), 3-hydroxyhexanoate (3HHx), and 4-hydroxybutyrate (4HB), with different ratios of monomers poured-solvent casting films and nanomembranes with oriented and non-oriented ultrathin fibers were obtained by electrostatic molding. With the use of SEM, AFM, and measurement of contact angles and energy characteristics, the surface properties and mechanical and biological properties of the polymer products were studied depending on the method of production and the composition of PHAs. It has been shown in cultures of mouse fibroblasts of the NIH 3T3 line and diploid human embryonic cells of the M22 line that elastic films and nanomembranes composed of P(3HB-co-4HB) copolymers have high biocompatibility and provide adhesion, proliferation and preservation of the high physiological activity of cells for up to 7 days. Polymer films, namely oriented and non-oriented nanomembranes coated with type 1 collagen, are positively evaluated as experimental wound dressings in experiments on laboratory animals with model and surgical skin lesions. The results of planimetric measurements of the dynamics of wound healing and analysis of histological sections showed the regeneration of model skin defects in groups of animals using experimental wound dressings from P(3HB-co-4HB) of all types, but most actively when using non-oriented nanomembranes obtained by electrospinning. The study highlights the importance of nonwoven nanomembranes obtained by electrospinning from degradable low-crystalline copolymers P(3HB-co-4HB) in the effectiveness of the skin wound healing process.

3D analyses reveal T cells with activated nuclear features in T-cell/histiocyte-rich large B-cell lymphoma

AbstractNodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) can show variable histological growth patterns and present remarkable overlap with T-cell/histiocyte-rich large B-cell lymphoma (THRLBCL). Previous studies suggest that NLPHL histological variants represent progression forms of NLPHL and THRLBCL transformation in aggressive disease. Since molecular studies of both lymphomas are limited due to the low number of tumor cells, the present study aimed to learn if a better understanding of these lymphomas is possible via detailed measurements of nuclear and cell size features in 2D and 3D sections. Whereas no significant differences were visible in 2D analyses, a slightly increased nuclear volume and a significantly enlarged cell size were noted in 3D measurements of the tumor cells of THRLBCL in comparison to typical NLPHL cases. Interestingly, not only was the size of the tumor cells increased in THRLBCL but also the nuclear volume of concomitant T cells in the reactive infiltrate when compared with typical NLPHL. Particularly CD8+ T cells had frequent contacts to tumor cells of THRLBCL. However, the nuclear volume of B cells was comparable in all cases. These results clearly demonstrate that 3D tissue analyses are superior to conventional 2D analyses of histological sections. Furthermore, the results point to a strong activation of T cells in THRLBCL, representing a cytotoxic response against the tumor cells with unclear effectiveness, resulting in enhanced swelling of the tumor cell bodies and limiting proliferative potential. Further molecular studies combining 3D tissue analyses and molecular data will help to gain profound insight into these ill-defined cellular processes.

Alternate day fasting and time-restricted feeding may confer similar neuroprotective effects during aging in male rats.

Age associated neurodegenerative changes are acknowledged to play a causative role in a majority of neurological diseases that accompany aging in organisms. To alleviate the deteriorative effects of aging in the brain, we investigated the effects oftwo types of intermittent fasting (IF) methods: alternate day fasting (ADF) and time- restricted feeding (TRF) in young (3months) and old (24months) in male Wistar rats comparing the results with age matched controls. The evaluation of biomarkers of oxidative stress showed significant decline in the old (ADF and TRF) groups in addition to up regulation in antioxidant levels. It was observed that ADF and TRF methods helped reduce ROS accumulation in the mitochondria and increased the activity of the electron transport chain complexes especially C-I and III. Gene expression analysis of autophagy genes like beclin and LC3B showed upregulated expression in ADF and TRF group. Sirtuin1 expression too significantly increased during fasting in both young and old groups showing fasting induced protection from aging. Histological analysis of sections of cerebral cortex and CA1 area provide evidence that fasting protected neurons against degeneration with age. Our results prompt us to conclude that the efficacy of these fasting methods ADF and TRF are reliable anti- aging strategies with respect to dietary restriction interventions. Moreover, both these methods compete closely in conferring protection from oxidative stress and inducing neuroprotective changes in brain of aged rats when compared to their young counterparts.

Real-time Analysis of Skin Biopsy Specimens With 2-Photon Fluorescence Microscopy

Nonmelanoma skin cancers (NMSCs) are primarily diagnosed through paraffin section histologic analysis of skin biopsy specimens that requires days to weeks before a formal diagnosis is reported. Two-photon fluorescence microscopy (TPFM) has the potential for point-of-care diagnosis of NMSC and other dermatologic conditions, which could enable same-visit diagnosis and treatment. To demonstrate that TPFM imaging of NMSC can occur within minutes of obtaining biopsies and provide similar histological features to those of conventional histology and evaluate TPFM diagnostic performance with respect to conventional histology. This comparative effectiveness pilot study examined 29 freshly excised biopsies from confirmed NMSC lesions in patients presenting for treatment. Biopsies underwent imaging immediately with TPFM on site at Rochester Dermatologic Surgery (Victor, New York) between October 2019 and August 2021. The imaged biopsies were subsequently submitted for paraffin histology to produce coregistered images. Twelve of these coregistered image pairs (41.4%) were used as a training set. Fifteen (51.7%) were used in a masked evaluation by a board-certified dermatopathologist. Two (6.9%) were excluded from the study before evaluation because they could not be coregistered. Sensitivity, specificity, and accuracy of TPFM for NMSC biopsies were evaluated compared with conventional histology. Fourteen of the 15 biopsy specimens (93.3%) in the evaluation set were identically diagnosed with TPFM and paraffin histology. The TPFM had 100% sensitivity (95% CI, 48%-100%), 100% specificity (95% CI, 69%-100%), and 100% accuracy (95% CI, 78%-100%) for basal cell carcinoma diagnosis. For squamous cell carcinoma diagnosis, TPFM had 89% sensitivity (95% CI, 52%-100), 100% specificity (95% CI, 54%-100%), and 93% accuracy (95% CI, 68%-100%). For overall NMSC diagnosis, TPFM had a 93% sensitivity (95% CI, 66%-100%), 100% specificity (95% CI, 3%-100%), and 93% accuracy (95% CI, 68%-100%). Examination of the 1 discordant pair revealed mismatched imaging planes as the source of error. The results of this comparative effectiveness pilot study suggest that TPFM captures histological characteristics of NMSC that are present in conventional histology, which reveals its potential as a rapid, point-of-care diagnostic alternative that does not need extensive sample preparation or retraining for image evaluation. Further validation of TPFM imaging performed for a larger cohort is needed to fully evaluate its diagnostic accuracy and potential effect within the field.