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Τρίτη 23 Φεβρουαρίου 2021

Pericyte morphology and function

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Histol Histopathol. 2021 Feb 17:18314. doi: 10.14670/HH-18-314. Online ahead of print.

ABSTRACT

The proper delivery of blood is essential for healthy neuronal function. The anatomical substrate for this precise mechanism is the neurovascular unit, which is formed by neurons, glial cells, endothelia, smooth muscle cells, and pericytes. Based on their particular location on the vessel wall, morphology, and protein expression, pericytes have been proposed as cells capable of regulating capillary blood flow. Pericytes are located around the microvessels, wrapping them with their processes. Their morphology and protein expression substantially vary along the vascular tree. Their contractibility is mediated by a unique cytoskeleton organization formed by filaments of actin that allows pericyte deformability with the consequent mechanical force transferred to the extracellular matrix for changing the diameter. Pericyte ultrastructure is chara cterized by large mitochondria likely to provide energy to regulate intracellular calcium concentration and fuel contraction. Accordingly, pericytes with compromised energy show a sustained intracellular calcium increase that leads to persistent microvascular constriction. Pericyte morphology is highly plastic and adapted for varying contractile capability along the microvascular tree, making pericytes ideal cells to regulate the capillary blood flow in response to local neuronal activity. Besides the vascular regulation, pericytes also play a role in the maintenance of the blood-brain/retina barrier, neovascularization and angiogenesis, and leukocyte transmigration. Here, we review the morphological and functional features of the pericytes as well as potential specific markers for the study of pericytes in the brain and retina.

PMID:33595091 | DOI:10.14670/HH-18-314

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Visual deficits after traumatic brain injury

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Histol Histopathol. 2021 Feb 18:18315. doi: 10.14670/HH-18-315. Online ahead of print.

ABSTRACT

Traumatic brain injury (TBI) is frequently described as any head injury ceasing the brain's normal function. Anatomically, developmentally, and physiologically, the eye is deemed as an extension of the brain. Vision in TBI is underrepresented, and the number of active clinical trials in this field are sparse. Frequently, visual problems are overlooked at the time of TBI, often resulting in progressive vision loss, lengthening, and impairing rehabilitation. TBI can be either penetrative or non-penetrative, associated with degeneration of neurons, apoptotic cell death, inflammation, microglial activation, hemorrhage associated with vascular dysfunction; however, precise animal modeling that mimics the extensive visual deficits of TBI pathology remain elusive. Recent works in both the diagnostics and therapeutics fields are starting to make sub stantial progress in the right direction. Discussion of current advancements in TBI animal models and the recent pathophysiological findings related to the neuro-glia-vascular unit (NVU) will help elucidate novel targets for potential therapeutics lines. Only over the past decade have newer pharmaceutical and stem cell-based treatments begun to come to light. The potency for these new lines of TBI specific curatives will be discussed along with the review of current blast-induced TBI models, providing potential directions for future research.

PMID:33599281 | DOI:10.14670/HH-18-315

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Biological and biocompatible characteristics of fullerenols nanomaterials for tissue engineering

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Histol Histopathol. 2021 Feb 19:18316. doi: 10.14670/HH-18-316. Online ahead of print.

ABSTRACT

Fullerenes, as hydrophobic molecules, are limited in biomedical function due to their very low solubility. But taking C60 (OH) x as an example, the properties of fullerenols were analyzed. It was found that fullerenols had good stability, water solubility, good biocompatibility and low cytotoxicity by adding a hydroxyl group to carbon atoms. In the biomedical field, it has been found that fullerene C60 can be used as a powerful free radical scavenger, with antioxidant activity, with antibacterial and inhibitory effects on cancer cells. Fullerenols inherit the good properties of fullerenes, and are better used in cancer treatment, including loading drug therapy and directly as an anticancer drug. In addition, fullerenols are also used in the repair of myocardial injury, the treatment of myocardial infarction and neuroprotection. With the deve lopment of tissue engineering technology, the preparation of nerve scaffolds which can improve ischemia, hypoxia and oxidative stress after nerve injury has become a research hotspot. The electron absorption and reduction characteristics of fullerenols in biomedical research bring new ideas for the treatment of oxidative stress in the repair of peripheral nerve defects. It seems that the research on fullerenols loaded neural scaffold has great prospects.

PMID:33604882 | DOI:10.14670/HH-18-316

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Optimisation and validation of immunohistochemistry protocols for cancer research

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Histol Histopathol. 2021 Feb 22:18317. doi: 10.14670/HH-18-317. Online ahead of print.

ABSTRACT

BACKGROUND: Immunohistochemistry (IHC) has become a valuable laboratory technique for diagnosing, evaluating metastasis and informing treatment selection in several cancers. Standardization however remains a limiting factor in IHC. The main aim of this research study was to optimise, validate and standardize antibodies and IHC protocols for cancer research.

METHODS: Seven monoclonal mouse and rabbit antibodies were optimised using formalin-fixed paraffin embedded (FFPE) human tissue blocks. 4um sections of FFPE block were stained using the Roche Ventana XT or Ventana ULTRA IHC automated analysers. This study modified manufacturer recommended protocols by using a unique antigen retrieval method, adding an amplification step, varying primary antibody incubation times, as well as using the Roche Ventana Ultraview detection system.

R ESULTS: Optimum antibody localisation was observed in modified IHC protocols in comparison with manufacturer recommended protocols for anti-CEACAM-1, anti-CD31, anti-COX-2, anti-HER-2/neu, anti-S100P, anti-thrombomodulin and anti-VEGFR-3. Majority of antibodies required more than one modification of the initial protocol. For anti-VEGFR-3 optimum staining was observed following 4 protocol modifications.

CONCLUSIONS: This study has optimised and standardized several tissue-based biomarkers that may be, in the future, used to screen, diagnose and monitor patients with certain cancer, such as bladder cancer. Accurate data on optimised protocols reduce time and resources wasted on experimental protocols, and ultimately help identify biomarkers or biomarker panels, which may be used to select treatment regimens for various cancers.

PMID:33616195 | DOI:10.14670/HH-18-317

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Morphological changes in mouse ovary due to hormonal hypersecretion and matrix metalloproteinase -2 activity

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Histol Histopathol. 2021 Feb 23:18318. doi: 10.14670/HH-18-318. Online ahead of print.

ABSTRACT

We analyzed whether aberrant gonadotropin secretion affects the morphological remodeling of murine ovarian tissues facilitated by activated matrix metalloproteinase (MMP) enzymes. Six mice were intraperitoneally injected with 5 IU of pregnant mare serum gonadotropin (PMSG) or human chorionic gonadotropin (HCG) every two days after estrus synchronization. Morphology and expression of various MMPs were assessed following the successful induction of hormonal secretion in these tissues. HCG treatment, but not PMSG treatment, resulted in the expanded production of granulose second follicular cells. In addition, the number of developing follicular cells in the HCG group increased compared with that in the PMSG group. Ovarian diameters were also very small in the PMSG group. Immunohistochemistry revealed decreased MMP-2 protein activity in the HCG group and increased MMP-2 activity in the PMSG group. Activity was particularly high in theca and granulose cells of the PMSG group, but only partial activity was observed in the theca cells of the HCG group. Vascular endothelial growth factor activity was increased in both the external and internal theca cell walls in the PMSG group while the HCG group showed high overall expression of this protein in the internal theca cells. These data indicate that follicular cell activity and remodeling of the ovaries differ based on the type of secretory hormone signals they receive. Inappropriate gonadotropin secretion may induce functional changes in the ovaries, and follicular remodeling may be facilitated by the activity of various MMPs.

PMID:33620082 | DOI:10.14670/HH-18-318

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Swabbing for SARS-CoV-2 in tracheostomized patients

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Via hno

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HNO. 2021 Feb 23. doi: 10.1007/s00106-021-01014-3. Online ahead of print.

NO ABSTRACT

PMID:33620506 | DOI:10.1007/s00106-021-01014-3

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The first year of the SARS-CoV-2 pandemic-impact on otorhinolaryngology

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Via hno

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HNO. 2021 Feb 23. doi: 10.1007/s00106-021-01015-2. Online ahead of print.

ABSTRACT

BACKGROUND: The first year of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has already affected our public health care system to an enormous extent and will continue to do so in the future. Otorhinolaryngologists (ORLs) are suspected to be at high risk of infection, due to the high viral load in the mucosa of the upper airways. The current review evaluates the impa ct of the pandemic on ORLs' activities and assesses the risk infection.

METHODS: A selective literature research was conducted using relevant English and German terms for ORL, SARS-CoV‑2, risk, and infection at PubMed, medRxiv, and bioRxiv, as well as in the Deutsches Ärzteblatt and on the websites of the Robert Koch Institute and the Johns Hopkins University.

RESULTS: Protection recommendations for ORL include general hygiene measures and wearing KN95 masks for routine professional activities. When in contact with coronavirus disease 2019 (COVID-19) patients, it is recommended to extend the personal protective equipment by eye protection, gloves, cap, and gown. International otorhinolaryngology societies have released guidelines for procedures (e.g., tracheostomy, sinus surgery), propagating personal protection for the surgical team and reduction of aerosols. Testing for SARS-CoV‑2 in patients and medical staff can contribute to reducing the risk of infection. Vacc ination would provide some additional protection for ORLs and other health care professionals with increased exposure to aerosols. There is increasing evidence that ORLs are at a high risk of contracting SARS-CoV‑2.

CONCLUSION: Consequent personal protection, frequent testing of patients and health care professionals, and the promised SARS-CoV‑2 vaccinations may provide adequate protection for highly exposed persons.

PMID:33620505 | DOI:10.1007/s00106-021-01015-2

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Telocytes in the atherosclerotic carotid artery: Immunofluorescence and TEM evidence

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Via histochem

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Acta Histochem. 2021 Feb;123(2):151681. doi: 10.1016/j.acthis.2021.151681. Epub 2021 Jan 22.

ABSTRACT

Telocytes, which possess distinct body shapes and long telopodes, are allocated in the vascular wall. As a fundamental cell type, telocytes construct a three-dimensional network to form a support structure for the artery. This study aims to characterize the morphology and ultrastructure of telocytes in atherosclerotic arteries. ApoE gene-deficient mice were selected as the a therosclerosis animal model and fed a high-fat diet for at least 12 weeks, and immunofluorescence assays and transmission electron microscopy techniques were used to observe changes in telocytes in atherosclerotic arteries. By immunofluorescence staining, CD34, CD117 and PDGFR-α were positive compared with negative CD28/vimentin in telocytes in the atherosclerotic carotid artery, and they were distributed in the tunica intima and tunica adventitia. Under transmission electron microscopy, the bodies of telocytes became larger, while telopodes became shorter compared with their normal condition, and a mass of lipidosomes was present during the progression of atherosclerosis. These results demonstrate that immunofluorescence with TEM is the critical method for identifying TCs and that steatosis of TCs is a reason for atherosclerotic artery dysfunction.

PMID:33493960 | DOI:10.1016/j.acthis.2021.151681

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Androgen and estrogen receptors immunolocalization in the sand rat (Psammomys Obesus) cauda epididymis

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Via histochem

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Acta Histochem. 2021 Feb;123(2):151683. doi: 10.1016/j.acthis.2021.151683. Epub 2021 Jan 25.

ABSTRACT

Both androgens and estrogens play key, albeit incompletely described, roles in the functioning of the epididymis. Because this tightly-coiled tubular structure is compartmented, precise mapping of the distribution of sex steroid's receptors is important. Such receptors have been located in the first segments (caput, corpus), but the last part (cauda) remains poorly explored. We used immunochemistry to localize androgen (AR) and estrogen (ESR1 and ESR2) receptors in the cauda in the fat sand rat (Psammomys obesus). We compared results obtained during the breeding versus resting seasons. We also used individuals castrated, or castrated then treated with testosterone, or subjected to the ligation of their efferent ducts. During the breeding season, in principal cells, we found strong staining both for AR and ESR1 in the apical cytoplasm, and strong staining for ESR2 in the nucleus. During the resting season, principal cells were positive for AR and ESR1, but negative for ESR2. In castrated animals, staining was null for ESR2 and AR, and weak for ESR1. In castrated then treated animals, immuno-expression was restored but only for AR and ESR1. Following efferent duct ligation, AR reactivity decreased while ESR1 and ESR2 provided strong staining. Broadly similar, but not fully identical patterns were observed in basal cells. They were positive for ESR2 and A R during the breeding season, but not for ESR1. During the resting season, staining was modest for ESR1 and AR and negative for ESR2. In all experimentally treated animals, we observed weak staining for AR and ESR1, and a lack of signal for ESR2. Overall, this study provides strong evidence that androgens and estrogens are involved in the seasonal regulation of the whole epididymis in the fat sand rat, with marked differences between caput and cauda (the corpus is highly reduced in rodent).

PMID:33508524 | DOI:10.1016/j.acthis.2021.151683

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The vomeronasal organ of the Korean black goat, Capra hircus coreanae

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Via histochem

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Acta Histochem. 2021 Feb;123(2):151684. doi: 10.1016/j.acthis.2021.151684. Epub 2021 Jan 28.

ABSTRACT

We examined the localization of olfactory marker protein (OMP), protein gene product9.5 (PGP9.5), and glycan diversity in the vomeronasal organ (VNO) of the Korean black goat (Capra hircus coreanae) during the prenatal and postnatal periods using immunohistochemistry and lectin histochemistry. In fetal and 1-day-old goats, OMP was occasionally identified in receptor cells of the VNO sensory epithelium, and PGP9.5 was localized in both the sensory and non-sensory epithelia. In VNO from adult goats, OMP was abundant in the sensory epithelium and scarce in single cells of the non-sensory epithelium. These results suggest that OMP production is initiated in the VNO sensory epithelium (VNE) during the fetal stage, and that its activity is increased in adult VNO receptor cells and solitary cells in the non-sensory epithelium (VNSE). Furthermore, the free borders of the sensory epithelia were positive for 7 lectins, and 6 lectins were moderately and/or highly abundant in receptor cells. Supporting and basal cells, and nerve bundles had similar expression patterns. In VNE, 7 lectins were observed in the free border, and 6 in ciliated, goblet, and basal cells, and in gland acini. The intensities of WGA, LCA, and PNA were high in VSE receptor cells, and the intensity of PNA was high in ciliated cells of the VNSE. The other 3 lectins showed similar patterns throu ghout development. Collectively, these results confirm that the Korean black goat VNO starts developing during the late fetal stages and differentiates further after birth.

PMID:33517140 | DOI:10.1016/j.acthis.2021.151684

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IGF-1 as selected growth factor multi-response to antidepressant-like substances activity in C57BL/6J mouse testis model

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Via histochem

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Acta Histochem. 2021 Feb 5;123(3):151685. doi: 10.1016/j.acthis.2021.151685. Online ahead of print.

ABSTRACT

Insulin-like growth factor (IGF-1) affects almost all cells in the body. Extremely important functions of this growth factor have been demonstrated in the brain and the reproductive system of both, females and males. Also, it is considered as a pro-inflammatory cytokine adjusting tissue homeostasis. However, it seems to play a special role in the male reproductive sys tem and it may be disturbed by the application of antidepressants with different mechanisms of drug action during therapy. To date, the effect of antidepressant-like substances (ALS) on the course of physiological processes in male testicular cells is poorly understood. Therefore, the purpose of the research was to determine the presence, localizationof IGF-1R (insulin-like growth factor 1 β receptor) and mRNA gene expression of IGF-1R and IGF-1 after administration of 3-[(2-methyl-1,3-tiazol-4-yl)ethynyl]-pyridine (MTEP) and N-[2-(Cyclohexyloxy)-4-nitrophenyl]-methanesulfonamide (NS-398) in the different scheme in the testis of mice. Imipramine was used as a reference drug having a documented interaction with the mGluR5 receptors. The immunohistochemical analyses showed the localization of IGF-1R in Sertoli, Leydig, and germinal cells after all used substances. Differences in receptor localization were observed depending on the drugs applied and the type of analyzed cells. In cont rast, there was a significant increase in IGF-1 gene expression after IMI + NS-398 and in IGF-1R after MTEP + NS-398 and IMI + NS-398 administration. It can, therefore, be assumed that the use of a combination of NS-398 with some ALS may run different mechanisms of action and affect the regulation of reproductive function in mouse testis through maintaining homeostasis at the molecular and immunological levels related to IGF.

PMID:33556704 | DOI:10.1016/j.acthis.2021.151685

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