Cells Lining Research Articles

Potential stem cell—Conditioned medium and their derived exosomes versus omeprazole in treatment of experimental model of gastric ulcer

Gastric ulcer is a common frequent clinical problem affecting all age and gender. This work aims to compare between the therapeutic effects of stem cell derived exosomes, stem cells conditioned medium and omeprazole on the healing of gastric ulcer model.Fifty rats were, assigned into 5 groups; control, gastric ulcer, omeprazole-treated, conditioned medium- treated, and exosomes-treated groups. Gastric ulcer was induced by aspirin dissolved in 1% carboxymethyl cellulose at a daily dose of 200 mg/kg for 5 consecutive days. Stomach specimens were obtained for histological, biochemical, and immunohistochemical assessments.The gastric ulcer group revealed widening of the fundic glands lumen containing, exfoliated dead cells. There was a remarkable distortion of the normal histological structure of the gastric mucosa with surface lining epithelial cell sloughing, vascular congestion and inflammatory cell infiltration. Both exosomes and conditioned medium treatments ameliorated almost all of the histopathological changes. Interestingly, the healing effect of exosomes was greater because it restored the histological architecture of gastric mucosa to nearly normal.In conclusion, this work may pave the future for using stem cell derived exosomes as a more convenient and effective adjuvant therapy in gastric ulcer.

The transcription factor PAX8 promotes angiogenesis in ovarian cancer through interaction with SOX17.

PAX8 is a master transcription factor that is essential during embryogenesis and promotes neoplastic growth. It is expressed by the secretory cells lining the female reproductive tract, and its deletion during development results in atresia of reproductive tract organs. Nearly all ovarian carcinomas express PAX8, and its knockdown results in apoptosis of ovarian cancer cells. To explore the role of PAX8 in these tissues, we purified the PAX8 protein complex from nonmalignant fallopian tube cells and high-grade serous ovarian carcinoma cell lines. We found that PAX8 was a member of a large chromatin remodeling complex and preferentially interacted with SOX17, another developmental transcription factor. Depleting either PAX8 or SOX17 from cancer cells altered the expression of factors involved in angiogenesis and functionally disrupted tubule and capillary formation in cell culture and mouse models. PAX8 and SOX17 in ovarian cancer cells promoted the secretion of angiogenic factors by suppressing the expression of SERPINE1, which encodes a proteinase inhibitor with antiangiogenic effects. The findings reveal a non-cell-autonomous function of these transcription factors in regulating angiogenesis in ovarian cancer.

Intravenous administration of umbilical cord lining stem cells in left ventricular assist device recipient: Rationale and design of the uSTOP LVAD BLEED pilot study

BackgroundLeft ventricular assist device (LVAD) implantation provides a robust survival advantage, however despite improvements in mortality, the adverse event burden of durable mechanical circulatory support remains high. Bleeding complications are one such significant complication. The uSTOP LVAD BLEED (Utilization of umbilical cord lining Stem cells TO Prevent LVAD associated angiodysplastic BLEEDing) pilot study is designed to evaluate the safety and tolerability of escalating doses of umbilical cord lining stem cells (ULSCs) in LVAD recipients to ameliorate the dysregulation of angiogenic factors seen in this population. DesignThis Phase Ia single-ascending dose pilot study will evaluate the IV administration of ULSCs in stable out-patients supported with an LVAD. In a 3 + 3 design, a maximum of 18 patients will receive an IV infusion of ULSCs. Main outcome measuresThe primary endpoints are safety and tolerability, secondary exploratory endpoints will include biomarker evaluation of angiogenic dysregulation. SummaryThis represents a novel cell type and route of administration in this population, while collecting initial data regarding the magnitude and duration of effects of cell therapy, and assessing the possibility of decreasing bleeding by a strategy of vascular stabilization. Clinical trial registrationClinicalTrials.gov Identifier: NCT04811261. https://clinicaltrials.gov/ct2/show/NCT04811261.

Loss of POGZ alters neural differentiation of human embryonic stem cells

POGZ is a pogo transposable element derived protein with multiple zinc finger domains. Many de novo loss-of-function (LoF) variants of the POGZ gene are associated with autism and other neurodevelopmental disorders. However, the role of POGZ in human cortical development remains poorly understood. Here we generated multiple POGZ LoF lines in H9 human embryonic stem cells (hESCs) using CRISPR/CAS9 genome editing. These lines were then differentiated into neural structures, similar to those found in early to mid-fetal human brain, a critical developmental stage for studying disease mechanisms of neurodevelopmental disorders. We found that the loss of POGZ reduced neural stem cell proliferation in excitatory cortex-patterned neural rosettes, structures analogous to the cortical ventricular zone in human fetal brain. As a result, fewer intermediate progenitor cells and early born neurons were generated. In addition, neuronal migration from the apical center to the basal surface of neural rosettes was perturbed due to the loss of POGZ. Furthermore, cortical-like excitatory neurons derived from multiple POGZ homozygous knockout lines exhibited a more simplified dendritic architecture compared to wild type lines. Our findings demonstrate how POGZ regulates early neurodevelopment in the context of human cells, and provide further understanding of the cellular pathogenesis of neurodevelopmental disorders associated with POGZ variants.

IL-6-MEDIATED OVEREXPRESSION OF HYBID (HYALURONAN-BINDING PROTEIN INVOLVED IN HYALURONAN DEPOLYMERIZATION) IN OSTEOARTHRITIC SYNOVIUM IS IMPLICATED FOR HYALURONAN DEGRADATION IN SYNOVIAL FLUIDS IN PATIENTS WITH KNEE OSTEOARTHRITIS

Purpose: Hyaluronan (HA)-binding protein involved in HA depolymerization (HYBID), alias cell migration-inducing protein (CEMIP) and KIAA1199, plays a key role in HA degradation. Although HYBID is expressed by rheumatoid synovial lining and sub-lining cells, little is known about cellular localization of HYBID in osteoarthritic synovial tissue. Transmembrane protein 2 (TMEM2), a transmembrane protein with sequence similarity to HYBID, is reportedly a hyaluronidase in mice. However, no studies have shown the expression or functional role of TMEM2 in human tissues.

The possible ameliorative role of Moringa oleifera seed oil on sofosbuvir-induced nephrotoxicity in albino rats; histopathological, immunohistochemical and biochemical studies

BackgroundHepatitis C is a liver infection caused by the hepatitis C virus (HCV). It can cause both acute and chronic hepatitis. Sofosbuvir (sofo) is a nucleotide analog inhibitor of HCV NS5B polymerase used to treat chronic hepatitis C infection as a component of a combination of antiviral treatment regimen. Many side effects of sofo were reported in different mammalian organs including kidney. Moringa oleifera (MO) is one of the medicinal plants which have many pharmacological activities and nutritional applications due to its rich phytonutrients content. This study aimed to investigate the possible ameliorative effect of MO seed oil against nephrotoxicity induced by sofo in adult male albino rats. The experimental animals were divided equally into four groups. Group I: animals were served as control. Group II: animals were orally given MO oil (2 ml/kg/day). Group III: animals were orally administered with sofo (36 mg/kg/day). Group IV: animals were orally given sofo then after 2 h they were given MO oil (with the same previous doses). All doses were daily given to the animals for eight weeks. At the end of the experiments, animals were sacrificed and sera were collected to determine urea, creatinine and malondialdehyde levels and catalase activity. Kidneys were removed out and prepared for both the histological and immunohistochemical studies.ResultsSofo-treated animals showed many pathological changes; damaged glomeruli and degenerated renal tubules with vacuolated lining epithelial cells contain pyknotic nuclei. In addition, leukocytic infiltration, congested blood vessels and hemorrhage were seen. Caspase-3 and PCNA were expressed in a large number of cells in the same group. Moreover, a significant increase in urea, creatinine and malondialdehyde levels was recorded as well as a significant decrease in catalase activity. Co-treatment of MO oil with sofo effectively counteracted the observed adverse effects. It attenuated the histological picture of the kidney, significantly ameliorated urea, creatinine and malondialdehyde levels and catalase activity and restored the normal expressions of caspase-3 and PCNA.ConclusionsMoringa oleifera oil can ameliorate nephrotoxicity induced by sofo via its antioxidant, anti-inflammatory and anti-apoptotic properties.

Patterns of changes in cellular composition of the stomach in chronic gastritis in children and adolescents

The aim of the work is to establish the regularities of changes in the cellular composition of the fundal and pyloric glands in chronic gastritis in children. Material and methods. 213 patients aged 7-17 years with chronic Hp-associated gastritis were examined. The cellular composition of the fundal and pyloric glands was determined in gastrobioptates. Immunohistochemical analysis was used to determine cells containing gastrin, somatostatin, pepsinogens I and II, acidic and neutral mucins. The results were expressed in‰ per 1000 epithelial cells. Results. As inflammation increases in the fundal glands, the number of main and cells containing pepsinogens I and II decreases, the number of lining, additional and endocrine cells increases. In the pyloric glands, the pool of endocrinocytes increases, the number of lining cells and cells immunopositive to pepsinogen II significantly decreases. Parallel to the severity of the process, there is a sharp increase in D- and G-cells and an increase in the number of epithelial cells containing acidic and neutral mucins. Conclusion. Chronic inflammation of the gastric mucosa is accompanied by a significant restructuring of the cellular composition of its glands.

Novel Blood Vascular Endothelial Subtype-Specific Markers in Human Skin Unearthed by Single-Cell Transcriptomic Profiling

Ample evidence pinpoints the phenotypic diversity of blood vessels (BVs) and site-specific functions of their lining endothelial cells (ECs). We harnessed single-cell RNA sequencing (scRNA-seq) to dissect the molecular heterogeneity of blood vascular endothelial cells (BECs) in healthy adult human skin and identified six different subpopulations, signifying arterioles, post-arterial capillaries, pre-venular capillaries, post-capillary venules, venules and collecting venules. Individual BEC subtypes exhibited distinctive transcriptomic landscapes associated with diverse biological pathways. These functionally distinct dermal BV segments were characterized by their unique compositions of conventional and novel markers (e.g., arteriole marker GJA5; arteriole capillary markers ASS1 and S100A4; pre-venular capillary markers SOX17 and PLAUR; venular markers EGR2 and LRG1), many of which have been implicated in vascular remodeling upon inflammatory responses. Immunofluorescence staining of human skin sections and whole-mount skin blocks confirmed the discrete expression of these markers along the blood vascular tree in situ, further corroborating BEC heterogeneity in human skin. Overall, our study molecularly refines individual BV compartments, whilst the identification of novel subtype-specific signatures provides more insights for future studies dissecting the responses of distinct vessel segments under pathological conditions.

IRIS: Infection with RespIratory Syncytial Virus in infants—a prospective observational cohort study

BackgroundRespiratory syncytial virus (RSV) is the most common cause of acute lower respiratory tract infection in infants. Globally, RSV is responsible for approximately 3.2 million hospital admissions and about 60,000 in-hospital deaths per year.MethodsInfection with RespIratory Syncytial Virus (IRIS) is an observational, multi-centre study enrolling infants with severe RSV infection and healthy controls. Inclusion criteria are age between 0 and 36 months and hospitalisation due to RSV infection at three German sites. Exclusion criteria are premature birth, congenital or acquired bronchopulmonary or cardiac diseases, and immunodeficiency. Healthy control probands are enrolled via recruitment of patients undergoing routine surgical procedures. Blood and respiratory specimens are collected upon admission, and RSV and other pathogens are analysed by multiplex polymerase chain reaction. Different biomaterials, including plasma, nasal lining fluid, blood cells, DNA, and RNA specimens, are sampled in a dedicated biobank. Detailed information on demographic characteristics and medical history is recorded, and comprehensive clinical data, including vital signs, medication, and interventions.DiscussionThe IRIS study aims to discover host and viral factors controlling RSV disease courses in infants. The approach including multi-omics characterisation in clinically well-characterized children with RSV bronchiolitis seeks to improve our understanding of the immune response against this virus. It may disclose novel diagnostic and treatment approaches for respiratory infections in infants.Trial registration: ClinicalTrials.gov, NCT04925310. Registered 01 October 2021—Retrospectively registered. https://clinicaltrials.gov/ct2/show/NCT04925310?cond=NCT04925310&draw=2&rank=1

Improved loss-of-function CRISPR-Cas9 genome editing in human cells concomitant with inhibition of TGF-β signaling

Strategies to modulate cellular DNA repair pathways hold immense potential to enhance the efficiency of CRISPR-Cas9 genome editing platform. In the absence of a repair template, CRISPR-Cas9-induced DNA double-strand breaks are repaired by the endogenous cellular DNA repair pathways to generate loss-of-function edits. Here, we describe a reporter-based assay for expeditious measurement of loss-of-function editing by CRISPR-Cas9. An unbiased chemical screen performed using this assay enabled the identification of small molecules that promote loss-of-function editing. Iterative rounds of screens reveal Repsox, a TGF-β signaling inhibitor, as a CRISPR-Cas9 editing efficiency enhancer. Repsox invariably increased CRISPR-Cas9 editing in a panel of commonly used cell lines in biomedical research and primary cells. Furthermore, Repsox-mediated editing enhancement in primary human CD4+ T cells enabled the generation of HIV-1-resistant cells with high efficiency. This study demonstrates the potential of transiently targeting cellular pathways by small molecules to improve genome editing for research applications and is expected to benefit gene therapy efforts.

The Effect of Whatsapp Group-Based Health Media on Knowledge, Attitudes and Behavior of Breast Cancer Early Detection of Health Students

Background: Breast cancer is a malignancy that develops from breast tissue that arises in the lining cells (epithelium) of the ducts or lobules. Breast cancer is one of the burdens of non-communicable diseases globally, increasing every year. Breast cancer is one of the most common cancers globally with 2,089 million new cases. One of the preventions of breast cancer that every woman can do is Breast Self-Examination (BSE). The objective of this study was to determine the effect of Whatsapp Group-based health promotion on the knowledge, attitudes, and behavior of early detection of breast cancer in health students in Badung Regency. Method:This study uses a cross-sectional research design with a total sample of 54 respondents. The study was conducted in one of the private universities in Badung Regency. Data were analyzed using SPSS, including univariate and bivariate analysis with the Wilcoxon signed-rank test. Result: The results found that the Whatsapp Group-based health promotion affected health students’ knowledge, attitudes, and behavior in Breast Self-Examination (BSE). This can be seen from the p-value of each variable, including knowledge (less than 0.001), attitude (less than 0.001), and behavior (less than 0.001).Conclusion: Counseling about BSE with health promotion media through Whatsapp Group is useful in increasing the knowledge, attitudes and behavior of health students

Current Management and Future Perspective in Pleural Mesothelioma.

Simple SummaryPlatin-based chemotherapy with pemetrexed has been the backbone of meaningful treatment in pleural mesothelioma for the last 16 years, alongside vinorelbine and gemcitabine, which show only modest response. Recently, a nivolumab and ipilimumab combination was approved, and it offers improvement in survival rate and quality of life. With the identification and understanding of novel biomarkers, many treatment options are currently being evaluated.Pleural mesothelioma is an aggressive malignancy arising from pleural mesothelial cell lining, predominantly associated with prior exposure to asbestos. The ban on asbestos use has led to its lower incidence in many countries, but globally the disease burden is expected to rise. Therefore, well-planned research is needed to develop more effective, tolerable and affordable drugs. The development of novel treatment has been too slow, with only two regimens of systemic therapy with robust phase 3 data approved formally to date. The treatment scenario for resectable disease remains controversial. However, recent developments in the understanding of disease and clinical trials have been encouraging, and may add better treatment options in the coming years. In this review, we discuss the current treatment options for pleural mesothelioma and shed light on some recent studies and ongoing trials.

Choosing the Reagent to Leach Fluorine from Spent Pot Lining of Aluminum Electrolysis Cells

The production of aluminum by electrolysis of cryolite-alumina melts is associated with generation of process waste including gas cleaning sludge, electric filter dust, coal flotation tailings, and spent pot lining (SPL). Most of this waste is then stored in sludge fields, deteriorating the environment near aluminum works. 30 to 50 kg of spent pot lining is produced per ton of Al. The authors’ research was dedicated to finding the best reagent for solvent extraction of valuable components (fluorine in particular) in the process of SPL hydrometallurgy. To that end, the research team sampled carbon parts of lining at the RUSAL Bratsk facility in Shelekhov, Irkutsk Oblast. The following reagents were tested: 2 wt.% solutions of sulfuric acid and sodium hydroxide. Leaching lasted 60 minutes in each experiment. 2% H2SO4solution was found to release fluorine at a higher rate. However, the resulting liquors contained sulfates, which might negatively affect the service life of equipment; for this reason, 2% NaOH solution was chosen for further leaching of fluorine from spent carbon parts of lining.

Effect of Zhuanggu Jianxi Decoction on Synovitis in Knee Osteoarthritis Rabbits via LXRs/NF-κB Pathway

<sec><title>Objective</title> To study the intervention effect and mechanism of Zhuanggu Jianxi decoction on synovitis in knee osteoarthritis (KOA) rabbits via the inflammatory regulation of synovial tissue and the expression of key node proteins in LXRs/NF-κB pathway. </sec><sec><title>Methods</title> Blank serum and drug-containing serum of rabbits were prepared for the intervention of synovial tissue. Twenty-two rabbits were randomly divided into two groups, with 16 in the model group and 6 in the blank group. The KOA model was injected with papain into the knee joint of the right hind limb. After the model was identified successfully, the synovial tissue of the right hind limb knee was cut into small pieces with a size about 1-2 mm³ under aseptic operation, 60 pieces were collected and randomly divided into model group, Zhuanggu Jianxi decoction group, LXRα inhibitor group and N-CoR inhibitor group with 15 pieces each, the synovial tissue pieces in each group were cultured in 6 well plates with every 3 pieces in each well, the four groups were intervened with 10% blank serum, 10% drug-containing serum, 10% drug-containing serum+5CPPSS-50 and 10% drug-containing serum+ML-792 respectively; the synovium of the blank group was prepared by the same method and cultured with 10% blank serum for 5 wells. After 7 days, synovial tissue was collected. Two wells of synovial tissue were taken from each group for HE staining, and the remaining synovial tissue was detected by Western blot and ELISA. </sec><sec><title>Results</title> 1) Morphology: compared with the blank group, HE staining in the model group showed the proliferation and disorder of synovial lining cells, a large number of inflammatory cell infiltration and fibrous tissue hyperplasia, and the morphological score was significantly increased (<italic>P</italic><0.05). Compared with the model group, mild hyperplasia and disorder of synovial lining cells, sparse scattered inflammatory cell infiltration and a small amount of fibrous tissue hyperplasia were observed in the Zhuanggu Jianxi decoction group, and the morphological score was significantly decreased (<italic>P</italic><0.05). Compared with the Zhuanggu Jianxi decoction group, the morphological score of pathological changes in the LXRα and N-CoR inhibitor groups was significantly increased (<italic>P</italic><0.05). 2) Inflammation-related indicators: compared with the blank group after intervention, the contents of IL-1β, TNF-α, MMP-3 and MMP-13 in the model group were significantly increased (<italic>P</italic><0.05); compared with the model group, the indexes above in the LXRα inhibitor group, N-CoR inhibitor group and Zhuanggu Jianxi decoction group were significantly decreased (<italic>P</italic><0.05); compared with the Zhuanggu Jianxi decoction group, the indexes above in the LXRα and N-CoR inhibitor groups were significantly increased (<italic>P</italic><0.05). Before and after intervention, the contents of IL-1β and TNF-α in the Zhuanggu Jianxi decoction group were significantly decreased (<italic>P</italic><0.05). 3) Protein expression: Compared with the blank group, the expressions of LXRα and N-CoR proteins in the model group were significantly decreased (<italic>P</italic><0.05), and the expression of P50 and P65 proteins were significantly increased (<italic>P</italic><0.05). Compared with the model group, LXRα and N-CoR protein in the LXRα inhibitor group and N-CoR protein in the N-CoR inhibitor group were not significantly decreased (<italic>P</italic>>0.05), P50 and P65 protein expression were significantly decreased (<italic>P</italic><0.05). The protein expression of LXRα and N-CoR in the Zhuanggu Jianxi decoction group were significantly increased (<italic>P</italic><0.05), and the protein expression of P50 and P65 were significantly decreased (<italic>P</italic><0.05). Compared with the Zhuanggu Jianxi decoction group, the protein expression of LXRα and N-CoR in both the LXRα and N-CoR inhibitor groups were significantly decreased (<italic>P</italic><0.05), and the protein expression of P50 and P65 were significantly increased (<italic>P</italic><0.05). </sec><sec><title>Conclusion</title> Zhuanggu Jianxi decoction could improve the pathological manifestations of synovial lining cell hyperplasia and the disorder arrangement, inflammatory cell infiltration and fibrous tissue hyperplasia, reduce the content of inflammatory related indexes in tissues, so as to reduce the inflammatory response of KOA synovial tissue in rabbits, which may be related to the up-regulation of LXRα and N-CoR protein expression in synovial cells via LXRs/NF-κB pathway in synovial tissue, thereby down-regulating P50 and P65 protein to reduce IL-1β, TNF-α, MMP-3 and MMP-13. </sec>

Orchestrated tumor apoptosis (Cu2+) and bone tissue calcification (Ca2+) by hierarchical Copper/Calcium-ensembled bioactive silica for osteosarcoma therapy

Malignant osteosarcoma is one of the serious health concerns attributed to inferior prognosis and uncontrolled metastasis in clinics. Indeed, multifunctional therapeutics with precise tumor killing and subsequent regeneration of damaged tissue are of immense requirement in treating osteosarcoma. Herein, this study demonstrates the fabrication of versatile calcium/copper-impregnated bioactive glass (Ca-BG/Cu-BG) frameworks with orchestrated cancer inhibition and bone tissue calcification towards osteosarcoma therapy. Initially, the designed composites are systematically characterized to explore various morphological attributes and physicochemical properties. The optimal formulation of Cu-BG (10 mol%) presented orchestrated effects of an excellent pH-responsive anticancer effect through hydroxyl radical (·OH) generation and facilitated hydroxyapatite formation towards bone tissue calcification. Further, in vitro investigations demonstrated substantial biocompatibility in mouse embryo osteoblast precursor cell line (MC3T3-E1) and exceptional anti-tumor effects in human osteosarcoma cells (HOS) through improved cellular internalization efficacy and free radical generation, respectively. Finally, in vivo immunostaining and micro-computed tomography (μCT) images of osteosarcoma-bearing mice confirmed Fenton-like reaction-induced chemodynamic therapy (CDT), over-expressed hypoxia-inducible factor (HIF)-1α through p53 pathway, and formation of calcified nodules. In addition, hematological analyses of hematoxylin-eosin (HE) stained main organ sections presented biosafety elucidations. Together, the synergistic prospects of Cu2+ and Ca2+ species had substantially orchestrated towards regenerative medicine.

Generation of a Cone Photoreceptor-specific GNGT2 Reporter Line in Human Pluripotent Stem Cells.

Fluorescent reporter lines generated in human pluripotent stem cells are a highly useful tool to track, isolate, and analyze cell types and lineages in live cultures. Here, we generate the first human cone photoreceptor reporter cell line by CRISPR/Cas9 genome editing of a human embryonic stem cell (hESC) line to tag both alleles of the Guanine nucleotide-binding protein subunit gamma-T2 (GNGT2) gene with a mCherry reporter cassette. Three-dimensional optic vesicle-like structures were produced to verify reporter fidelity and track cones throughout their development in culture. The GNGT2-T2A-mCherry hESC line faithfully and robustly labels GNGT2-expressing cones throughout the entirety of their differentiation in vitro, recapitulating normal fetal expression of this gene. Our observations indicate that human cones undergo significant migratory activity during the course of differentiation in vitro. Consistent with this, our analysis of human fetal retinae from different stages of development finds positional differences of the cone population depending on their state of maturation. This novel reporter line will provide a useful tool for investigating human cone development and disease.

Caffeine docking studies with keratin: Implications for its cosmetic applications

Keratin is found in hair, nails, horn, hoofs, and wool, as well as the epithelial cells of the skin's outer layers. Keratin is a structural and protective protein found in the epithelium. Some keratins also regulate biological functions like cell proliferation and protein synthesis. Keratins help build hair, nails, and the skin's outer layer. They are present on the lining of organs, glands, and other bodily cells. In this work, we have given theoretical evidence to emphasize the role of caffeine, in hair growth and cosmetic applications (skin) through docking studies. Auto Dock and Auto Dock Tools version 4.2 were used to dock molecules. The results show, higher keratin binding affinity of caffeine (ΔG −4.3 kcal/mol) suggests that caffeine could involve in hair growth formulations and skin care applications which make them suitable for cosmetic application. The reported data requires in vitro and in vivo validation.

NHLRC2 expression is increased in idiopathic pulmonary fibrosis

BackgroundVariants of NHL repeat-containing protein 2 (NHLRC2) have been associated with severe fibrotic interstitial lung disease in early childhood and NHLRC2 has been listed as a differentially expressed gene between rapidly and slowly progressing idiopathic pulmonary fibrosis (IPF) patients. However, its cell type-specific localization in human lung tissue is unknown. The aim of this study was to evaluate NHLRC2 mRNA and protein expression in different cell types of lung tissue samples and to investigate the effect of transforming growth factor (TGF)-β1 exposure on NHLRC2 expression in vitro.MethodsThe NHLRC2 expression in lung tissue samples was studied by immunohistochemistry (50 IPF, 10 controls) and mRNA in situ hybridization (8 IPF, 3 controls). The immunohistochemical NHLRC2 expression was quantified with image analysis software and associated with the clinical and smoking data of the patients. NHLRC2 expression levels in primary stromal and small airway epithelial cell lines after exposure to TGF-β1 was measured by quantitative reverse transcription polymerase chain reaction and Western blot analysis.ResultsNHLRC2 expression was detected especially in bronchiolar epithelial cells, type II pneumocytes and macrophages in normal lung. In the lungs of IPF patients, NHLRC2 was mainly expressed in hyperplastic alveolar epithelial cells lining fibroblast foci and honeycombs. NHLRC2 expression assessed by image analysis was higher in IPF compared to controls (p < 0.001). Ever-smokers had more prominent NHLRC2 staining than non-smokers (p = 0.037) among IPF patients. TGF-β1 exposure did not influence NHLRC2 levels in lung cell lines.ConclusionsNHLRC2 expression was higher in IPF compared to controls being widely expressed in type II pneumocytes, macrophages, bronchiolar epithelium, and hyperplastic alveolar epithelium. Additionally, its expression was not regulated by the exposure to TGF-β1 in vitro. Further studies are needed to clarify the role of NHLRC2 in IPF.

MORPHOLOGICAL JUSTIFICATION OF THE SIGNIFICANCE OF LUMBAR ARTERIES IN THE PREVENTION OF SPINAL CORD ISCHEMIA IN ATHEROSCLEROTIC LESIONS OF ABDOMINAL AORTA.

The aim: To investigate atherosclerosis of the abdominal part of the aorta and atherosclerosis of the lumbar arteries, aimed at improving surgical tactics during reconstructive interventions on the abdominal part of the aorta in patients with multifocal atherosclerosis. Materials and methods: 20 autopsies were performed. The macro preparation consisted of the part of the abdominal aorta 1х1 sm taken along with a separated lumbar artery for 1-1,5 sm. Histological cuts were coloured with hematoxylin and eosin. The histochemical research was conducted in order to establish changes in all layers of blood vessels. The immunohistochemical research was carried out along with generally accepted histological methods in 10 cases to determine the morphological vessel wall functional state, connective tissue and smooth muscle components. Results: In all cases there were morphological signs of aortic atherosclerosis with different degrees of damage to the lumen. In none of the analyzed cases did we establish pathomorphological signs of atherosclerosis of the lumbar artery distal to the mouth. In the walls of the lumbar arteries, we noted the preservation of the layered structure, the integrity of the endothelial cells, the normal arrangement of smooth muscle cells. Conclusions: It was established that there cannot be the atherosclerotic occlusion of the lumbar artery, since there are no signs of atheromatous lesions in its walls, the structure of all layers is preserved, the endothelial damage is absent, the location and structure of the lining cells is normal.

Transit along the vas deferens results in a high percentage of filiform spermatozoa with compacted chromatin in the rooster (Gallus domesticus).

The present work aimed to evaluate the chromatin compaction of rooster spermatozoa along the male reproductive tract, and to study the vas deferens lining cells, potentially involved in sperm maturation. Chromomycin A3 (CMA3) was used to determine the chromatin compaction of spermatozoa from testis (T), proximal (including epididymis, V1), intermediate (V2) and distal (V3) vas deferens, and ejaculate (E). Six Birchen Leonesa roosters were used. E was obtained in vivo by dorso-ventral massage. V1, V2 and V3 sperm were obtained post mortem (six pairs of vasa deferentia), by flushing. T was obtained by washing the testes, cut in halves. The fixed cells were stained with CMA3 and propidium iodide for flow cytometry assessment. Results showed higher (P P P.