scholarly journals Comparison of cryoprotective methods for histological examination of rat and porcine lung tissue

2021 ◽  
Vol 90 (2) ◽  
pp. 225-231
Author(s):  
Rea Jarošová ◽  
Petra Ondráčková ◽  
Zdeněk Patočka ◽  
Zbyšek Sládek
Keyword(s):  
Lung Tissue ◽  
Immunohistochemical Analysis ◽  
Supercooled Liquids ◽  
Histological Evaluation ◽  
Pulmonary Tissue ◽  
Pulmonary Parenchyma ◽  
Target Molecules ◽  
Phosphate Buffered Saline ◽  
Tissue Cryopreservation ◽  
Tissue Reactivity

Proper histological evaluation of lung tissue and accurate antigen detection by immunodetection techniques require histological tissue processing to preserve tissue reactivity and open alveolar spaces. In this study, we focused on testing and comparing different procedures of tissue cryopreservation. Sucrose or Tissue Tek were used with several methods of freezing samples by supercooled liquids and liquid nitrogen. Changes in tissue caused during the freezing of samples and the effect of cryoprotectants on the tissue were recorded. Rat and porcine pulmonary tissues were used in this experiment. This study aimed to optimize the process of lung cryopreservation with emphasis on enabling proper anatomical evaluation and preserving a high tissue immunoreactivity. The best results were obtained by inflating pulmonary parenchyma with a 1 : 1 mixture of O.C.T. with phosphate buffered saline (PBS) frozen in supercooled n-heptane placed on dry ice. Pulmonary tissue prepared in this way enables to perform proper histological evaluation and to detect target molecules by immunohistochemical analysis.

scholarly journals Intestinal Anti-Inflammatory Activity of the Aqueous Extract from Ipomoea asarifolia in DNBS-Induced Colitis in Rats

2018 ◽  
Vol 19 (12) ◽  
pp. 4016 ◽  
Author(s):  
Valéria da Silva ◽  
Aurigena de Araújo ◽  
Daline Araújo ◽  
Maíra Lima ◽  
Roseane Vasconcelos ◽  
...  
Keyword(s):  
Protective Effect ◽  
Aqueous Extract ◽  
Intestinal Inflammation ◽  
Activity Index ◽  
Immunohistochemical Analysis ◽  
Histological Evaluation ◽  
Myeloperoxidase Activity ◽  
Down Regulation ◽  
Anti Inflammatory ◽  
Ipomoea Asarifolia

Inflammatory bowel disease is triggered by an uncontrolled immune response associated with genetic, environmental, and intestinal microbiota imbalance. Ipomoea asarifolia (IA), popularly known as “salsa” or “brave salsa”, belongs to the Convolvulaceae family. The aim of this approach was to study the preventive effect of IA aqueous extract in 2,4-dinitrobenzene sulfonic acid (DNBS)-induced colitis in rats. Rats pretreated with IA extract or sulfasalazine (SSZ) received intracolonic instillation of DNBS in 50% ethanol (v/v). IA extract presented a protective effect against intestinal inflammation, with improvement in the disease activity index and macroscopic damage. IA or SSZ significantly reduced myeloperoxidase activity, and also down-regulation of the gene expression of JNK1, NF-κβ-p65, STAT3, and decreased levels of TNFα, IL-1β, and increased IL-10, associated with a significant improvement of oxidative stress, in addition to a reduction in MDA and an increase of glutathione in colonic tissue. The protective effect of the extract was also confirmed in histological evaluation, showing preservation of the colonic cytoarchitecture. Immunohistochemical analysis revealed down-regulation of NF-κβ-p65, iNOS, IL-17, and up-regulation of SOCs-1 and MUC-2. IA extract presents antioxidant and anti-inflammatory intestinal properties, and proved to be a potential application for preventing damage induced by DNBS.

scholarly journals Effects of CX3CL1 inhibition on murine bleomycin-induced interstitial pneumonia

2020 ◽  
Vol 18 ◽  
pp. 205873922095990
Author(s):  
Soichi Yamada ◽  
Shion Miyoshi ◽  
Junko Nishio ◽  
Satoshi Mizutani ◽  
Zento Yamada ◽  
...  
Keyword(s):  
Interstitial Pneumonia ◽  
T Cell Activation ◽  
Lung Tissue ◽  
Cell Activation ◽  
Immunohistochemical Analysis ◽  
Inflammatory Cells ◽  
Lavage Fluid ◽  
Novel Therapy ◽  
Tissue Sections

Background: Treatment for interstitial pneumonia (IP) associated with collagen diseases has not been established. There is a need to elucidate the pathogenesis of IP and develop a novel therapy. We aimed to clarify the role of chemokine (C-X3-C motif) ligand 1 (CX3CL1, also known as fractalkine) in IP. Methods: Bleomycin (BLM) was intratracheally administered to C57BL/6 mice to induce IP. For treatment with control Ab or anti-CX3CL1 mAb, the mice were administered either Ab three times per week for 2 weeks from the day of BLM administration until euthanasia. Expressions of CX3CL1 and its unique receptor CX3CR1 in the lung tissue were examined by immunohistochemical analysis. Cellular infiltration and lung fibrosis were evaluated based on hematoxylin-eosin-staining and Sirius red staining of the lung tissue sections, respectively. Bronchoalveolar lavage fluid (BALF) cells were analyzed by flow cytometry. Results: CX3CL1 and CX3CR1 were strongly expressed in the lung tissue from mice with BLM-induced IP (BLM-IP). Treatment with anti-CX3CL1 mAb did not significantly alter inflammatory cell infiltration or fibrosis in the lung tissue. However, the number of M1-like macrophages in BALF was decreased and surface CD3 expression on T cells was increased by anti-CX3CL1 mAb treatment. Conclusions: Inhibition of CX3CL1 decreased inflammatory cells and may attenuate T cell activation in BALF. CX3CL1 inhibitor may have the potential to suppress the infiltration and activation of immune cells in IP.

Invasive pulmonary aspergillosis in a child with acute myeloid leukaemia: pharmacotherapy and surgical management

Oncoreview ◽  
2016 ◽  
Vol 6 (4) ◽  
pp. 0-0
Author(s):  
Jan Styczyński
Keyword(s):  
Acute Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Surgical Management ◽  
Lung Tissue ◽  
Invasive Pulmonary Aspergillosis ◽  
Invasive Fungal Disease ◽  
Pulmonary Tissue ◽  
Hematopoietic Stem ◽  
Good Penetration ◽  
Acute Myeloid

This paper reports on diagnostic and therapeutic management of pulmonary invasive fungal disease (IFD) in a child with relapsed acute myeloid leukaemia, undergoing chemotherapy followed by hematopoietic stem cell transplantation. Surgical management with resection of the involved lung tissue was based on the location of fungal infiltrates close to large circulatory vessels. After examination of resected pulmonary tissue, a diagnosis of proven IFD was done. This case report is an example that aspergillosis is usually the cause for pulmonary IFD. Pharmacotherapy of pulmonary IFD should be based on compounds with good penetration to lung tissue: amphotericin B lipid form or voriconazole.

The air cysts and cystoid changes in pulmonary tissue

2020 ◽  
Vol 29 (6) ◽  
pp. 745-754
Author(s):  
M. A. Karnaushkina ◽  
D. V. Burenchev ◽  
A. D. Strutynskaya
Keyword(s):  
Lung Tissue ◽  
Clinical Symptoms ◽  
Lung Parenchyma ◽  
Diagnostic Methods ◽  
Pulmonary Tissue ◽  
Clinical Observations ◽  
Bronchopulmonary Diseases ◽  
Tissue Changes ◽  
Radiological Pattern ◽  
Radiological Patterns

Computed tomography (CT) of chest organs is one of the most accurate diagnostic methods allowing the physician to assess the condition of lung parenchyma. Correct interpretation of CT results requires the clinician to recognize normal appearance of lung parenchyma on X-ray and know changes visualized in various bronchopulmonary diseases. It is important that the physician knows and understands underlying cause of a particular radiological pattern in order to discuss with the radiologist lung tissue changes that have been identified considering clinical symptoms. Descriptions of radiological patterns and discussion of corresponding typical clinical observations are presented in the article devoted to air cyst syndrome and cystoid changes in the lung tissue.

BCL3 and NSCLC: Are they related?

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e18507-e18507
Author(s):  
Fotinos-Ioannis D Dimitrakopoulos ◽  
Anastasia E Kottorou ◽  
Anna G Antonacopoulou ◽  
Stella Marousi ◽  
Ioulia Koukourikou ◽  
...  
Keyword(s):  
Lung Tissue ◽  
Immunohistochemical Analysis ◽  
Stage Ii ◽  
Disease Stage ◽  
Normal Lung Tissue ◽  
Maximum Diameter ◽  
Normal Lung ◽  
Histological Subtype ◽  
Healthy Donors ◽  
Significant Difference

e18507 Background: NFkB pathways have become objects of detailed research in the last years, although, little is known of the possible role of BCL3 in lung carcinogenesis. The aim of this study was to define the relation of the BCL3 single nucleotide polymorphism rs8100239 with NSCLC and its association with BCL3 protein expression. Methods: Using Tagger and Sysnps programs, we chose the rs8100239 polymorphism of BCL3.We used 294 blood and FFPE normal tissue specimens from patients with NSCLC and 280 blood specimens from healthy donors. DNA isolation was performed using the Qiagen DNA blood and the QIAamp DNΑ FFPE Tissue kits. Samples were genotyped using real-time PCR. Immunohistochemical analysis for BCL3 was performed on 90 FFPE specimens (80 tumors and 10 tumor-adjacent normal tissues) from NSCLC patients. Results: Approximately half of the healthy donors (45.5%) were AT heterozygotes, 36.6% were TT and 17.9% were AA homozygotes. The respective frequencies in patients were 43.8%, 33.1% and 23.1%. There was no statistically significant difference in allele frequencies between healthy controls and patients (p=0.297). However, patients of stage II carrying a T allele displayed two and five-year survival benefit compared to patients with AA genotype (p<0.001). Immunohistochemical analysis for BCL3 protein revealed a statistically significant difference between malignant and normal lung tissue (p<0.001). More specifically, BCL3 was detected in both the cytoplasm and nucleus in neoplastic tissues with strong intensity, whereas in non neoplastic tissue no immunostaining was noticed. BCL3 protein levels were not associated with the polymorphism. However, cytoplasmic expression of BCL3 was correlated with age (p=0.009), histological subtype (p=0.031) and disease stage (0.038). Furthermore, tumors with smaller diameter had higher BCL3 nuclear expression levels than larger tumors (p=0.009). Conclusions: Patients of stage II carrying a T allele had improved two and five-year survival. BCL3 expression is not correlated with rs8100239 status. However, it is related to histological subtype, stage and maximum diameter while it differs between malignant and normal lung tissue. This research has been co-financed by the EU and Greek national funds (Heracleitus II Program).

scholarly journals Ex Vivo Artifacts and Histopathologic Pitfalls in the Lung

2016 ◽  
Vol 140 (3) ◽  
pp. 212-220 ◽  
Author(s):  
Erik Thunnissen ◽  
Hans J. L. G. Blaauwgeers ◽  
Erienne M. V. de Cuba ◽  
Ching Yong Yick ◽  
Douglas B. Flieder
Keyword(s):  
Smooth Muscle ◽  
Lung Tissue ◽  
Ex Vivo ◽  
Open Lung ◽  
Pulmonary Parenchyma ◽  
Lung Biopsies ◽  
Diagnostic Pitfalls ◽  
Morphologic Appearance

Context Surgical and pathologic handling of lung physically affects lung tissue. This leads to artifacts that alter the morphologic appearance of pulmonary parenchyma. Objective —To describe and illustrate mechanisms of ex vivo artifacts that may lead to diagnostic pitfalls. Design In this study 4 mechanisms of ex vivo artifacts and corresponding diagnostic pitfalls are described and illustrated. Results —The 4 patterns of artifacts are: (1) surgical collapse, due to the removal of air and blood from pulmonary resections; (2) ex vivo contraction of bronchial and bronchiolar smooth muscle; (3) clamping edema of open lung biopsies; and (4) spreading of tissue fragments and individual cells through a knife surface. Morphologic pitfalls include diagnostic patterns of adenocarcinoma, asthma, constrictive bronchiolitis, and lymphedema. Conclusion Four patterns of pulmonary ex vivo artifacts are important to recognize in order to avoid morphologic misinterpretations.

scholarly journals Possibility of detecting “middle-mass molecules” in the pulmonary tissue of fatally injured persons

2013 ◽  
Vol 17 (3 (67) p.1) ◽  
pp. 16-17
Author(s):  
A. M. Biliakov
Keyword(s):  
Lung Tissue ◽  
Forensic Medicine ◽  
Early Stage ◽  
Tissue Extract ◽  
Pulmonary Tissue ◽  
Quantitative Content ◽  
Short Period

In order to solve the problem whether it is possible to use “middle-mass molecules” for the purposes of forensic medicine, their content has been measured in a lung tissue extract of persons who died at an early stage of the antemortem period. It has been found out that “middle-mass molecules” are detected in an extract of the lung tissue of persons who died immediately after an injury, within a short period of time (from several minutes to some tens of minutes) and in 1 to 2 hours after being injured. Moreover, their quantitative content in those who died 1 to 2 hours after the injury is statistically different from other groups of injured persons (р<0,05).

Lung tissue volume during development of edema in isolated canine lungs

1980 ◽  
Vol 48 (6) ◽  
pp. 1038-1044 ◽  
Author(s):  
C. R. Felton ◽  
W. G. Johanson
Keyword(s):  
Pulmonary Edema ◽  
Blood Volume ◽  
Lung Tissue ◽  
Venous Pressure ◽  
Base Line ◽  
Abrupt Increase ◽  
Lung Weight ◽  
Pulmonary Tissue ◽  
Tissue Volume ◽  
Pulmonary Tissue Volume

The technique of estimating pulmonary tissue volume (Vt) by rebreathing a tissue-soluble gas is rapid and noninvasive. We examined the sensitivity of this technique for the estimation of Vt in isolated, perfused canine lungs during the development of pulmonary edema. Vt was 84 ± 15% (mean ± SD) of the associated lung weight for lung weights of up to 240% of base line but decreased to 70 ± 2% when lung weight exceeded 310% of base line. Small rebreathing tidal volumes resulted in significantly smaller values for Vt in edematous lungs. An abrupt increase in pulmonary venous pressure increased lung weight due to vascular distension; Vt measurements detected less than half of this increase, implying that certain portions of the intravascular blood volume are not measured by this technique.

Microvascular fluid flow in ex vivo and engineered lungs

2021 ◽  
Author(s):  
Micha Sam Brickman Raredon ◽  
Alexander James Engler ◽  
Yifan Yuan ◽  
Allison Marie Greaney ◽  
Laura E. Niklason
Keyword(s):  
Fluid Flow ◽  
Ex Vivo ◽  
Mechanistic Model ◽  
Culture Fluid ◽  
Pulmonary Tissue ◽  
Capillary Recruitment ◽  
Pulmonary Parenchyma ◽  
Barrier Resistance ◽  
Microvascular Recruitment ◽  
Insight Into

In recent years, it has become common to experiment with ex vivo perfused lungs for organ transplantation, and to attempt regenerative pulmonary engineering using decellularized lung matrices. However, our understanding of the physiology of ex vivo organ perfusion is imperfect: it is not currently well understood how decreasing microvascular barrier affects the perfusion of pulmonary parenchyma. Additionally, protocols for lung perfusion and organ culture fluid-handling are far from standardized, with widespread variation on both basic methods and on ideally controlled parameters. To address both of these deficits, a robust, non-invasive, and mechanistic model is needed which is able to predict microvascular resistance and permeability in perfused lungs while providing insight into capillary recruitment. Although validated mathematical models exist for fluid flow in native pulmonary tissue, previous models generally assume minimal intravascular leak from artery to vein and do not assess capillary bed recruitment. Such models are difficult to apply to both ex vivo lung perfusions, in which edema can develop over time and microvessels can become blocked, and to decellularized ex vivo organomimetic cultures, in which microvascular recruitment is variable and arterially-perfused fluid enters into the alveolar space. Here, we develop a mathematical model of pulmonary microvascular fluid flow which is applicable in both instances, and we apply our model to data from native, decellularized, and regenerating lungs under ex vivo perfusion. The results provide substantial insight into microvascular pressure-flow mechanics, while producing previously unknown output values for tissue-specific capillary-alveolar hydraulic conductivity, microvascular recruitment, and total organ barrier resistance.

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