Q112363514
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Vyd. 1. 106 s. : il., tab. ; 30 cm
Cílem skripta je podat základní informace zejména o struktuře a ultrastruktuře živočišných buněk s přihlédnutím k jejich funkci.
- Konspekt
- Buněčná biologie. Cytologie
- Učební osnovy. Vyučovací předměty. Učebnice
- NLK Obory
- biologie
- cytologie, klinická cytologie
- NLK Publikační typ
- učebnice vysokých škol
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.
The objective of the present study was to compare the effects of dietary oils containing polyunsaturated fatty acids (PUFA) n-3 and n-6, respectively, on cutaneous wound healing in rats, and to demonstrate the usefulness of the matrix-assisted-laser-desorption-ionization mass spectroscopy-imaging (MALDI MSI) method in this type of experiment. Superiority of PUFA n-3 in this context was the tested hypothesis. Four groups of male Wistar rats by twelve animals each were fed a diet with added 5% of palm oil (P; control), fish oil (F), Schizochytrium microalga oil (Sch) and safflower oil (S), respectively, for eight weeks. Consequently, dorsal full-thickness cutaneous excisions were performed, and selected markers of wound healing were evaluated 18 days post excision. The median of signal intensity corresponding to an amount of collagen α-1 (III) fragment, quantified using MALDI MSI, decreased in a sequence P > F > Sch > S (P < 0.001). Using haematoxylin-eosin staining of the histological preparations, semi-quantitatively assessed epithelium height tended to decrease in the order of P > S > Sch > F; the wound extent in the sequence of P > Sch > S > F; and the sequence of the progress of neo-angiogenesis was assessed as S > P > F ≈ Sch. It was concluded that the tested hypothesis was confirmed only partially: PUFA n-3 showed better results regarding the wound extent, but were inferior in terms of epithelium height and progress of neo-angiogenesis. This was the first time MALDI MSI was successfully employed for evaluating skin wound healing in a rat model.
- MeSH
- dietní tuky nenasycené * terapeutické užití MeSH
- dietní tuky terapeutické užití MeSH
- hojení ran * MeSH
- kolagen typu I, řetězec alfa 1 analýza MeSH
- nenasycené mastné kyseliny terapeutické užití MeSH
- potkani Wistar MeSH
- rány a poranění dietoterapie veterinární MeSH
- reepitalizace MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice statistika a číselné údaje veterinární MeSH
- výsledek terapie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- klinická studie MeSH
- MeSH
- buněčná diferenciace účinky léků MeSH
- buněčné linie MeSH
- cytokiny genetika metabolismus MeSH
- interleukin-10 genetika metabolismus MeSH
- interleukin-6 genetika metabolismus MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- lipopolysacharidy farmakologie MeSH
- makrofágy metabolismus účinky léků MeSH
- nádory prostaty * MeSH
- TNF-alfa genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Lactobacillus, Bifidobacterium and Enterococcus cultures are increasingly used as probiotics for humans and pigs. The aim of this study was to investigate if co-cultivation of porcine and human neutrophils with probiotics can lead to increased apoptosis in vitro. Ten adult Large white pigs and 10 healthy human donors were used in this study. Neutrophils were isolated by dextran sedimentation and cultivated with and without the lactic acid bacteria Bifidobacterium bifidum, Lactobacillus rhamnosus, and Enterococcus faecium for 2, 4, 24 and 48 h. Early and late apoptosis was measured using flow cytometry, and cell lysis was detected based on lactate dehydrogenase activity (LDH). A significant (P < 0.05; P < 0.01) increase in apoptotic neutrophils and LDH was observed at 24 h and 48 h in vitro. All probiotics exerted their greatest effects on the early apoptosis of porcine neutrophils, while the effects of L. rhamnosus were most pronounced on late apoptosis and those of B. bifidumon LDH release of human neutrophils. The increased neutrophil apoptosis caused by probiotic bacteria can be beneficial for more efficient efferocytosis and faster resolution of inflammation and tissue regeneration. In conclusion, we have demonstrated that the interaction of B. bifidum, L. rhamnosus,and E. faecium with human and porcine neutrophils leads to their apoptosis.
- MeSH
- apoptóza MeSH
- Bifidobacterium bifidum fyziologie imunologie MeSH
- Enterococcus faecium fyziologie imunologie MeSH
- Lacticaseibacillus rhamnosus fyziologie imunologie MeSH
- lidé MeSH
- neutrofily MeSH
- probiotika * aplikace a dávkování farmakologie metabolismus MeSH
- průtoková cytometrie metody MeSH
- Sus scrofa MeSH
- techniky in vitro MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
BACKGROUND: Macrophages may play a prominent role in defense of the bovine mammary gland, and their functionality is necessary for successful eradication of bacterial pathogens. In contrast to necrosis, however, apoptosis has not yet been studied in macrophages from bovine mammary glands. Therefore, the aim of this study was to confirm the occurrence of apoptosis in macrophages from resting heifer mammary glands and during the inflammatory response. METHODS: Inflammatory response was induced by phosphate buffered saline (PBS) and by lipopolysaccharide (LPS). Resident macrophages (RESMAC) were obtained before and inflammatory macrophages (INFMAC) 24, 48, 72 and 168 hours after inducing inflammatory response in mammary glands of unbred heifers. Cell samples were analyzed for differential counts, apoptosis and necrosis using flow cytometry. RESULTS: Populations of RESMAC and INFMAC contained monocyte-like cells and vacuolized cells. Apoptosis was detected differentially in both morphologically different types of RESMAC and INFMAC and also during initiation and resolution of the inflammatory response. In the RESMAC population, approximately one-tenth of monocyte-like cells and one-third of vacuolized cells were apoptotic. In the INFMAC population obtained 24 h after PBS treatment, approximately one-tenth of monocyte-like cells and almost one-quarter of vacuolized cells were apoptotic. At the same time following LPS, however, we observed a significantly lower percentage of apoptotic cells in the population of monocyte-like INFMAC and vacuolized INFMAC. Moreover, a higher percentage of apoptotic cells in INFMAC was detected during all time points after PBS in contrast to LPS. Comparing RESMAC and INFMAC, we observed that vacuolized cells from populations of RESMAC and INFMAC underwent apoptosis more intensively than did monocyte-like cells. CONCLUSIONS: We conclude that apoptosis of virgin mammary gland macrophages is involved in regulating their lifespan, and it is involved in the resolution process of the inflammatory response.
- MeSH
- apoptóza MeSH
- kultivované buňky MeSH
- lipopolysacharidové receptory metabolismus MeSH
- lipopolysacharidy farmakologie MeSH
- makrofágy cytologie metabolismus MeSH
- mléčné žlázy zvířat cytologie účinky léků patologie MeSH
- regulace genové exprese MeSH
- skot MeSH
- viabilita buněk fyziologie MeSH
- zánět chemicky indukované MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of the study was to evaluate the effect of selected temperatures on viability (apoptosis and necrosis) of bovine blood neutrophil granulocytes (neutrophils) in vitro. The following temperatures were tested: -80, -20, 4, 23, 37 degrees C. Heparinised bovine blood was incubated for 1, 4 and 24 h under respective temperature. Apoptosis and necrosis of neutrophils were detected by light microscopy, transmission electron microscopy (TEM) and flow cytometry (FCM). From selected temperatures, 4 degrees C impaired the neutrophil viability least. The proportion of apoptotic and necrotic neutrophils amounted to (mean +/- SD) 5.25 +/- 3.53% and 0.83 +/- 0.38%; 7.09 +/- 2.07% and 1.64 +/- 0.50%; 35.39 +/- 12.53% and 5.46 +/- 1.46%; after 1, 4 and 24 h incubation, respectively. The temperature (4 degrees C) is the best alternative for short-term storage.
- MeSH
- apoptóza MeSH
- experimentální radiační poranění MeSH
- financování organizované MeSH
- kryoprezervace metody veterinární MeSH
- nekróza patologie MeSH
- neutrofily patologie MeSH
- počet leukocytů MeSH
- průtoková cytometrie MeSH
- skot MeSH
- teplota MeSH
- transmisní elektronová mikroskopie MeSH
- viabilita buněk MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
