Valproate is known to disturb the kidney function, and high doses or prolonged intake may cause serum ion imbalance, kidney tubular acidosis, proteinuria, hyperuricosuria, polyuria, polydipsia, and ...dehydration. The aim of this
study was to see whether naringin would counter the adverse effects of high-dose valproate in C57Bl/6 mice and to which extent. As expected, valproate (150 mg/kg bw a day for 10 days) caused serum hyperkalaemia, more in male than female mice. Naringin reversed (25 mg/kg bw a day for 10 days) the hyperkalaemia and activated antioxidative defence mechanisms (mainly catalase and glutathione), again more efficiently in females. In males naringin combined with valproate was not as effective and even showed some prooxidative effects.
Gallic acid has been identified as an antioxidant component of the edible and medicinal plant Peltiphyllum peltatum. The present study examined its potential protective role against sodium fluoride ...(NaF)-induced oxidative stress in rat erythrocytes. Oxidative stress was induced by NaF administration through drinking water (1030.675 mg m-3 for one week). Gallic acid at 10 mg kg
and 20 mg kg
and vitamin C for positive controls (10 mg kg
) were administered daily intraperitoneally for one week prior to NaF administration. Thiobarbituric acid reactive substances, antioxidant enzyme activities (superoxide dismutase and catalase), and the level of reduced glutathione were evaluated in rat erythrocytes. Lipid peroxidation in NaF-exposed rats significantly increased (by 88.8 %) when compared to the control group (p<0.05). Pre-treatment with gallic acid suppressed lipid peroxidation in erythrocytes in a dose-dependent manner. Catalase and superoxide dismutase enzyme activities and glutathione levels were reduced by NaF intoxication by 54.4 %, 63.69 %, and 42 % (p<0.001; vs. untreated control group), respectively. Pre-treatment with gallic acid or vitamin C significantly attenuated the deleterious effects. Gallic acid isolated from Peltiphyllum peltatum and vitamin C mitigated the NaF-induced oxidative stress in rat erythrocytes.
Galna kiselina u ranijim je istraživanjima potvrđena kao važna antioksidativna sastavnica jestive i ljekovite biljke Peltiphyllum peltatum. U ovome istraživanju ispitan je njen potencijalni zaštitni učinak protiv oksidativnog stresa izazvanog natrijevim fluoridom (NaF) u eritrocitima štakora. Oksidativni stres izazvan je davanjem NaF eksperimentalnim životinjama putem vode za piće (1030,675 mg m
) tijekom jednog tjedna. Galna kiselina životinjama je davana intraperitonealno u dozama 10 mg kg
i 20 mg kg
na dan. Životinje iz pozitivno kontrolne skupine svaki su dan, osim NaF, intraperitonealno primale vitamin C u dozi 10 mg kg
. U eritrocitima štakora primjenom TBARS-testa izmjerene su razine lipidne peroksidacije, aktivnosti antioksidativnih enzima (superoksid dismutaze i katalaze) te razina reduciranoga glutationa. Lipidna peroksidacija u štakora izloženih NaF osjetno se povećala (88,8 %) u usporedbi s kontrolnom skupinom (p<0,05). Prethodni tretman galnom kiselinom smanjio je razinu lipidne peroksidacije u eritrocitima, ovisno o dozi. Toksičnost NaF smanjila je aktivnost enzima katalaze i superoksid dismutaze i razine glutationa za 54,4 %, 63,69 %, odnosno 42 % (p<0,001; u usporedbi s kontrolnom skupinom). Dobiveni rezultati upućuju na pozitivne učinke predobrade štakora galnom kiselinom izoliranom iz Peltiphyllum peltatum i vitaminom C na razinu oksidativnog stresa izazvanog natrijevim fl uoridom u eritrocitima štakora.
General anaesthetics are often used in patients who are under oxidative stress due to a critical illness or surgical trauma. Some anaesthetics may worsen oxidative stress and some may act as ...antioxidants. The aim of this study was to evaluate liver, brain, kidney, and lung tissue oxidative stress in rats exposed to desflurane and sevoflurane and in unexposed rats. The animals were divided in three groups: control (received only air); sevoflurane (8 %), and desflurane (4 %). After four hours of exposure, we evaluated the levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), Cu, and Zn. Exposure to either of the anaesthetics significantly increased lung MDA levels compared to control (Mann-Whitney U test; P<0.05), probably because it is the tissue directly exposed to anaesthetic gases. Oxidative stress and antioxidant activity in other tissues varied between the desflurane and sevoflurane groups. Our results suggest that anaesthesiologist should not only be aware of the oxidative or antioxidative potential of anaesthetics they use, but should also base their choices on organs which are the most affected by their oxidative action.
Poznato je da kirurške zahvate prati porast razine oksidativnog stresa, kako zbog stvaranja slobodnih kisikovih radikala tako i zbog smanjene aktivnosti obrambenih sustava koji se mogu oduprijeti njihovu djelovanju. Stoga su saznanja o antioksidativnom kapacitetu anestetika koji se primjenjuju prije nekoga kirurškog zahvata vrlo važna i od velikog su kliničkog značenja. Sevofluran i desfluran su inhalacijski anestetici koji se učestalo rabe u svrhu uvođenja bolesnika u anesteziju. Cilj ovog istraživanja bio je utvrditi razine oksidativnog stresa u različitim tkivima štakora i usporediti razlike u odgovoru tkiva na izlaganje navedenim anesteticima. U tu svrhu razine oksidativnog stresa izmjerili smo u jetri, mozgu, bubrezima i plućima štakora podijeljenih u tri eksperimentalne skupine. Kontrolna skupina udisala je samo zrak, dok su druge dvije skupine izložene 8 %-tnomu sevofluranu te 4 %-tnomu desfluranu tijekom 4 h. Nakon završetka obrade životinje su žrtvovane i uzimani su im uzorci tkiva za biokemijske analize. Mjerena je razina malondialdehida (MDA), aktivnost enzima superoksid dismutaze (SOD) i glutation peroksidaze (GSH-Px) te razine bakra i cinka. Izloženost anesteticima izazvala je oksidativni stres u plućima, na što upućuje značajno povišena razina MDA (Mann-Whitney U-test P<0.05) izmjerena u plućnom tkivu štakora obiju izloženih skupina u odnosu na kontrolu. Plućno je tkivo u odnosu na ostala tkiva podložnije štetnim utjecajima reaktivnih kisikovih radikala vjerojatno stoga što je ono prvo izloženo plinovitim anesteticima nakon njihova ulaska u organizam. Razine oksidativnog stresa i antioksidativne aktivnosti koje smo izmjerili u ostalim tkivima bile su različite te su ovisile o primijenjenom anestetiku. Na osnovi dobivenih rezultata možemo zaključiti da bi se zbog različitog odgovora tkiva izbor anestetika trebao provoditi na individualnoj osnovi.
Uvod/cilj: Hemodijaliza (HD) je povezana s brojnim biokemijskim abnormalnostima uključujući dislipidemiju i oksidativni stres. Cilj istraživanja bio je ispitati aktivnost i utvrditi povezanost enzima ...superoksid dismutaze (SOD) s parametrima lipidnog profi la kod bolesnika na HD. Metode: Presječnom studijom obuhvaćeno je 100 HD bolesnika koji su podijeljeni u dvije skupine: bolesnici koji su bili liječeni pomoću HD više od tri mjeseca, ali manje od pet godina (HD <5 godina, n=48) i bolesnici koji su bili liječeni pomoću HD pet godina i više (HD ≥5 godina, n=52). Kontrolnu skupinu (n=50) činile su po dobi i spolu odgovarajuće zdrave osobe bez subjektivnih i objektivnih pokazatelja bubrežne bolesti. Uzorci krvi služili su za određivanje koncentracija SOD, ukupnog kolesterola, triglicerida (TG), lipoproteina male gustoće (LDL) i lipoproteina velike gustoće (HDL). Serumska koncentracija SOD određena je metodom ELISA upotrebom komercijalnog kita. Rezultati: Serumska koncentracija SOD bila veća kod HD bolesnika u usporedbi sa zdravim kontrolnim osobama. Koncentracija SOD u skupini bolesnika na HD <5 godina bila je značajno veća u odnosu na kontrolnu skupinu ispitanika 12,29 (10,85-14,15 prema 11,04 (9,42-12,99) U/mL (p<0,05), ali nije utvrđena značajna razlika u koncentraciji SOD između skupina bolesnika na HD ≥5 godina 12,97 (10,27-14,56) U/mL i zdravih ispitanika kontrolne skupine. Uz to, nije bilo značajne razlike u serumskoj koncentraciji SOD između HD bolesnika koji su bili liječeni pomoću HD <5 godina i ≥5 godina. Utvrđeno je značajno smanjenje koncentracija TC, LDL i HDL u objema skupinama HD bolesnika u odnosu na kontrolnu skupinu (p<0,0005), ali razine serumskih TG nisu se značajno razlikovale između promatranih skupina. Rezultati dobiveni ROC analizom pokazali su da su serumske razine TC, LDL i HDL u odnosu na serumsku koncentraciju SOD osjetljivije u razlikovanju HD bolesnika i zdravih osoba. Zaključak: Na osnovi naših rezultata može se zaključiti da povišena serumska koncentracija SOD, zajedno s promjenama lipidnog profi la, povećava rizik od ateroskleroze i kardiovaskularnih komplikacija kod HD bolesnika.
Provider: - Institution: - Data provided by Europeana Collections- During the life cycle aerobic organisms are exposed to a number of endogenous
and exogenous sources of ROS. Present in low to ...moderate concentrations these
reactive molecules play an important role in many physiological processes
such as regulation of signaling cascades and gene expression. In high
concentrations, ROS can oxidize cellular proteins, lipids and DNA, causing
changes in structure and function, damage and even cell death. When the
concentration of free radicals exceeds the physiological level, a cell is
said to be in a state of oxidative stress. To prevent the onset and reduce
the consequences of oxidative stress, living organisms have developed
powerful antioxidant system (AOS). This system includes a set of mechanisms
to maintain the level of free radicals in the narrow range between
physiological and toxic concentrations. The most important enzyme components
in AOS are: superoxide dismutase (SOD), catalase (CAT), glutathione
peroxidase (GPx) and glutathione reductase (GR). These enzymes are involved
in the removal of ROS and participate in maintaining level of reduced
glutathione, thus ensuring the preservation of physiological functions and
inhibit the development of cell damage and disease. Because of such an
important role of AOS, the regulation of the functional expression of its
components is of particular importance for physiological and pathological
processes in aerobic organisms. One of the most important regulatory
molecules in this system is Nrf2 (NF-E2 related factor 2). Nrf2 is a
transcription factor that induces expression of many cytoprotective proteins
including antioxidant enzymes, and therefore play an important role in the
regulation of oxidative stress. Previous studies have clearly demonstrated a
link between oxidative stress and carcinogenesis. Cancer is a multistage
process that develops over three stages: initiation, promotion and
progression and oxidative stress is associated with each of them. Moreover,
antioxidant profile is altered in cancer cells compared with healthy, normal
tissue, so these molecules may be important biomarkers in the assessment of
risk and the degree of carcinogenesis. Endometrial cancer is one of the three
most common diseases of female reproductive organs. It has been shown that
benign and premalignant changes precede the malignant transformation of the
uterus, which is why these conditions may be considered as stages of
carcinogenesis. Despite numerous studies, the molecular processes involved in
multi-stage development of endometrial cancer are not yet fully known.
Therefore, the aim of this dissertation was to examine the changes in
expression of the four most important antioxidant enzymes in the blood and
tissues of patients with benign, premalignant and malignant endometrial
transformations, as well as the mechanism of their regulation by
transcription factor Nrf2. In this research we used endometrial tissue and
venous blood of patients diagnosed with: polypus endometrii, uterus
myomatosus, hyperplasia simplex endometrii, hyperplasia complex endometrii
and adenocarcinoma endometrii. Polypus endometrii and uterus myomatosus were
considered as benign, while hyperplasia simplex and hyperplasia complex
endometrii were considered as premalignant transformation of the uterus.
After adequate sample preparation, we initially determined the total protein
concentration in blood and endometrial tissue of all five groups of patients.
Then we performed SDS PAGE electrophoresis, transfer to nitrocellulose
membrane and chemiluminescent detection of CuZnSOD, CAT, GPx, GR and Nfr2
protein molecules. The protein levels were normalized with respect to
β-actin. Furthermore, in endometrial samples of all five groups of patients,
RNA was extracted and underwent reverse transcription into cDNA, followed by
the TaqMan Real-Time PCR method by which the amounts of CuZnSOD, CAT, GPx, GR
and Nfr2 mRNA were determined in respect to POLR2 as endogenous control. The
results of these experiments showed the following: 1. In comparison to
patients with polyps and myomas, in endometrium of women with hyperplasia
simplex and hyperplasia complex decreased levels of Nrf2 were recorded, while
in adenocarcinoma tissue Nrf2 protein level was increased. The observed
changes were regulated at the transcriptional level, except in adenocarcinoma
tissue in which increase in Nrf2 level was regulated by some
post-transcriptional mechanism. 2. CuZnSOD mRNA level and the amount of
CuZnSOD protein in the endometrium of patients with hyperplasia simplex and
hyperplasia complex were decreased compared to patients with polyps and
myomas. In adenocarcinoma tissue, however, the values of these parameters
were significantly increased compared to both benign and both premalignant
transformations. Observed transcriptional and translational CuZnSOD
variability in different stages of endometrial transformation were the
consequences of changes in the level of transcription factor Nrf2. 3.
Compared to the control groups with polyps and myomas, levels of CAT mRNA and
CAT protein were significantly decreased in the tissues of patients with
premalignant and malignant lesions of the uterus. It is also shown that Nrf2
had no direct effect on the CAT gene transcription, but acting on other
translational and posttranslational processes can probably influence the
level of this enzyme in the endometrium. 4. During carcinogenesis, increase
of GPx protein level was detected in premalignant endometrium, while decline
in the the amount of this enzyme was measured in adenocarcinoma tissue. GPx
level was positively correlated with the amount of transcription factor Nrf2,
but mechanism that regulates the expression of this enzyme varied with the
degree of tissue transformation. 5. In the course of malignant endometrial
transformation, increased expression of glutathione reductase was observed.
Consequent increased level of GR protein was the result not only of the
intensified transcription under the influence of Nrf2, but also of an
additional, posttranscriptional mechanism activated in transformed cells of
the uterus. 6. As regards AOE expression in the blood of examined patients,
it was observed that in comparison to the control group, in groups of women
with benign endometrial changes a decline in Nrf2 and GR levels were
measured, GPx level was increased, while the relative amounts of CuZnSOD and
CAT protein did not change significantly. Somewhat more pronounced changes
were observed in the blood of patients diagnosed with hyperplasia simplex,
hyperplasia complex and adenocarcinoma, characterized by declined protein
levels of Nrf2, CuZnSOD and GR, increased level of GPx and unaltered
expression of CAT. It was also shown that the blood expression levels of all
investigated AOE correlated positively with the amount of Nrf2. The results
of this dissertation indicate the existence and importance of the specific
expression pattern of antioxidant enzymes and transcription factor Nrf2 in
gynecological patients diagnosed with: polypus endometrii, uterus myomatosus,
hyperplasia simplex endometrii, hyperplasia complex endometrii and
adenocarcinoma endometrii. The observed changes confirm the important role of
the examined enzymes in complex molecular interactions that underlie the
transformation of endometrial cells. It is obvious that these AO molecules
not only influence the development of lesions in the uterine tissue, but they
are also an important factor for the progression of benign changes in
premalignant and malignant phenotype. It can be concluded that the results of
this dissertation contribute significantly to a better understanding of the
molecular processes involved in carcinogenesis. Implementation of this
knowledge in clinical practice could contribute to the quality of prevention,
diagnosis and treatment of gynecological patients, may lead to to improvement
in the disease course and prognosis and, consequently, may result in
increased survival rate of cancer patients.- Tokom čitavog životnog ciklusa aerobni organizmi su izloženi brojnim
endogenim i egzogenim faktorima koji indukuju povećanje produkcije ROS-a.
Prisutni u fiziološkim koncentracijama, reaktivni molekuli ROS-a imaju
značajnu ulogu u važnim ćelijskim procesima kao što su regulacija signalnih
kaskada i genske ekspresije. U visokim koncentracijama ROS mogu oksidovati
ćelijske proteine, lipide i DNK i time dovesti do promena strukture i
funkcije, oštećenja, pa i smrti ćelije. Kada koncentracija slobodnih radikala
premaši fiziološki nivo, smatra se da se ćelija nalazi u stanju oksidativnog
stresa. Kako bi sprečili nastanak i umanjili posledice ove vrste stresa, živi
organizmi su razvili moćan antioksidativni sistem zaštite (AOS). Ovaj sistem
uključuje seriju mehanizama kojima se nivo slobodnih radikala održava u uskom
opsegu između fiziološke i toksične koncentracije. Najvažnije enzimske
komponente AOS su: superoksid dismutaze (SOD), katalaza (CAT), glutation
peroksidaza (GPx) i glutation reduktaza (GR). Ovi enzimi uklanjaju višak
ROS-a i učestvuju u održavanju nivoa redukovanog glutationa, čime obezbeđuju
očuvanje osnovnih životnih funkcija i sprečavaju nastanak oštećenja i bolesti
ćelije. Zbog tako važne uloge AOS, regulacija funkcionalne ekspresije
njegovih komponenti je naročito značajna za fiziološke i patološke procese u
aerobnim organizmima. Jedan od najvažnijih regulatornih molekula u tom
sistemu je Nrf2 (NF-E2 related factor 2). Nrf2 je transkripcioni faktor koji
indukuje ekspresiju mnogih citoprotektivnih proteina uključujući i
antioksidativne enzime, zbog čega ima značajnu ulogu u regulaciji
oksidativnog stresa. Savremena istraživanja na različitim model-sistemima
ukazuju na povezanost oksidativnog stresa i kancerogeneze. Razvoj kancera je
višestepeni proces koji se razvija kroz tri faze: inicijaciju, promociju i
progresiju, a oksidativni stres je povezan sa svakim od ova tr
Provider: - Institution: - Data provided by Europeana Collections- Dental pulp originates from the embryonic ectomesenchyme and represents potentially important source of mesenchymal stem cells ...(MSCs) with the ability to regenerate all tissues of the craniofacial region, originating from the ectomesenchyme, too. N-acetyl-L-cysteine (NAC) is an antioxidant that may have an impact on the therapeutic use of MSCs. The aim of this study was to determine whether the cells isolated from the dental pulp of children deciduous teeth of children and expanded in vitro, have the characteristics of MSC, to examine the effect of NAC on their proliferation and differentiation and to determine whether NAC influences the metabolism of glucose, the activity of antioxidant enzymes and whether it reduces oxidative damage of various cellular macromolecules. The initial cell population was obtained from six pulps of deciduous teeth using ex vivo tissue explants method. The number of colonies (Colony Forming Unit – Fibroblast; CFU-F) was determined by seeding the low density cell culture. Proliferative potential and population doubling time of the cells in culture were followed by counting the cells after tripsinization of subconfluent cultures, every 4 days, respectively, during 40 days of cultivation. Flow cytometry was used for cell immunophenotypisation, aldehyde dehydrogenase (ALDH) activity determination, number of cells in different phases of cell cycle, number of cell divisions and percentage of cells in apoptosis. Activity of β-galactosidase (SA-β-Gal), was determined using cytochemistry. Osteogenesis, chondrogenesis and adipogenesis were induced using complete commercial mediums. Osteogenic differentiation was monitored via alkaline phosphatase activity (spectrophotometry), deposition of calcium in the extracellular matrix (Alizarin red staining) and the appearance of osteocalcin (immunocytochemistry). Chondrogenic differentiation was followed by measuring collagen type I (in situ hybridization), collagen type 2 (immunohistochemistry) and the determination of the concentration of glycosaminoglycans (spectrophotometry). Adipogenic differentiation was examined by visualization of fat droplets (Oil Red O staining). Activity of catalase and superoxide dismutase (SOD) in cell lysates was determined spectrophotometrically. Malondialdehyde (MDA) was determined by reaction with thiobarbituric acid (TBA). Carbonyl derivatives of proteins were determined by reaction with 2,4-dinitrophenylhydrazine. After total lipid extraction and methylation of fatty acids, fatty acid methyl esters were separated by gas-liquid chromatography, and the concentration of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids was expressed as percentage of total fatty acids detected in the sample. Isoenzyme forms of lactate dehydrogenase (LDH1, LDH2, LDH3, LDH4 and LDH5) were determined by vertical electrophoresis. All the above measurements were made with and without different concentrations of NAC (0,1 mM, 1 mM and 2 mM). Our results showed that the dental pulp of deciduous teeth of children had population of cells that formed a significant number of CFU-F (4% of the isolated cells) and continually proliferated for 40 days without decrease in the population doubling time. However, the resulting cell population was heterogeneous in terms of the division velocity. After three days of cultivation, around 12% of the cells were divided less than four times, while the remaining cells divided more than four times. Viability of cells was in average 95%, and only 3% of the cells were positive for β-galactosidase. During the entire time of the cultivation, expanded cells retained the ability to differentiate into osteoblasts, adipocytes and chondrocytes. Almost all cells expressed CD44, CD73, collagen type I, CD29, CD90 and osteonectin. Part of the cells expressed STRO-1, CD146 and CD106. Marker of hematopoietic stem cells have not been detected. Elevated ALDH activity ranged from 4% to 24% and was negatively correlated with the population doubling time. Our results showed that the effect of NAC on isolated cells dependeds on its concentration. In fact, after 72 hours of application of lower NAC concentrations (0,1 mM and 1 mM) the number of cells in culture increased by 20%, most probably due to the acceleration of the cell cycle of a small population of cells. Application of 2 mM NAC decreased the number of cells in culture. In addition, the growing concentration of NAC stimulated osteogenesis and chondrogenesis and inhibited adipogenesis. Also, the application of 0,1 mM NAC increased the percentage of cells which expressed high ALDH activity, and reduced the percentage of SA-β-gal positive cells. Application of 2 mM NAC led to an increase in the number of SA-β-gal positive cells. Lower concentrations of NAC reduced oxidative damage to various cellular macromolecules (reduced lipid peroxidation and the formation of carbonyl derivatives of proteins), and this was accompanied by reduced activity of SOD and catalase. In addition, it was shown that by adding 0,1 mM NAC the activity of LDH isoforms 4 and 5 increased, which could indicate an increase in the glycolitic activity, and at least in part explain the rapid proliferation of cells. After a comprehensive review of our results, we could conclude that by the method we applied, we isolated heterogeneous population of cells from the pulp of children deciduous teeth. Isolated cells had characteristics of dental pulp stem cells. Lower concentrations of NAC stimulated proliferation of cells in culture, reducing their oxidative damage and possibly increasing the volume of glycolysis, while all concentrations of NAC stimulated osteogenesis and chondrogenesis. We believe that the low concentration of NAC could find application in dental pulp stem cell expansion protocols, esspecially if necessary to stimulate their differentiation toward osteoblasts and chondrocytes.- Zubna pulpa vodi poreklo od embrionalnog ektomezenhima i potencijalno je važan izvor mezenhimalnih matičnih ćelija (MMĆ) za regeneraciju svih tkiva kraniofacijalne regije koja su, takođe, poreklom od ektomezenhima. N-acetil-L-cistein (NAC) je antioksidant koji može da ima uticaj na terapijsku primenu MMĆ. Cilj ovog rada je bio da utvrdi da li ćelije izolovane iz zubne pulpe mlečnih zuba dece i ekspandirane in vitro, imaju karakteristike MMĆ i da ispita dejstvo NAC-a na njihovu proliferaciju i diferencijaciju, kao i da se utvrdi da li NAC utiče na aktivnost enzima antioksidativne zaštite, oksidativno oštećenje različitih ćelijskih struktura i metabolizam glukoze. Metodom tkivnog eksplanta, iz šest pulpi mlečnih zuba dece, je dobijena početna populacija adheretnih ćelija. Za procenu broja CFU-F (eng. Colony Forming Unit – Fibroblast), ćelije su zasejavane u maloj gustini. Proliferativni potencijal i vreme udvajanja broja ćelija u kulturi je praćeno brojanjem ćelija posle tripsinizacije subkonfluentnih kultura, svaka 4 dana, tokom 40 dana. Protočna citometrija je korišćena za imunofenotipizaciju ex vivo umnoženih ćelija, određivanje aktivnosti aldehid- dehidrogenaze (ALDH), brzinu ulaska ćelija u sintetsku (S) fazu ćelijskog ciklusa, određivanje broja ćelijskih deoba i detekciju apoptoze. Aktivnost -galaktozidaze (SA-β-Gal), određivana je korišćenjem citohemije. Diferencijacija ekspandiranih ćelija u smeru osteogeneze, hondrogeneze i adipogeneze izvedena je korišćenjem komercijalnih medijuma. Promene na ćelijama tokom osteogene diferencijacije su praćene preko aktivnosti alkalne fosfataze spektrofotometrijski, deponovanja kalcijuma u ekstracelularni matriks (bojenje alizarin crvenim) i pojave osteokalcina (imunocitohemija). Hondrogena diferencijacija u peletama je praćena određivanjem kolagena tip I (in situ hibridizacija), kolagena tip 2 (imunohistohemija) i određivanjem koncentracije glikozaminoglikana (spektrofotometrija). Adipogena diferencijacija je ispitana vizuelizacijom masnih kapljica (Oil red O bojenje). Aktivnost katalaze i superoksid-dismutaze (SOD) u ćelijskom lizatu je određena spektrofotometrijski. Koncentracija malondialdehida (MDA) je određena reakcijom sa tiobarbiturnom kiselinom (TBA). Karbonilni derivati proteina određivani su reakcijom sa 2,4 -dinitrofenilhidrazinom. Posle ekstrakcije ukupnih lipida i metilovanja masnih kiselina, metil-estri masnih kiselina su razdvajani gasno-tečnom hromatografijom (GLC), a koncentracija zasićenih (SFA), mononezasićenih (MUFA) i i polinezasićenih masnih kiselina (PUFA) je izražena procentualno u odnosu na ukupne masne kiseline. Izoenzimski oblici laktat-dehidrogenaze (LDH1, LDH2, LDH3, LDH4 i LDH5) su odreĊivani vertikalnom elektroforezom. Sva navedena merenja su izvedena bez i sa različitim koncentracijama NAC-a (0,1 mM, 1 mM i 2 mM). Naši rezultati su pokazali da je iz zubne pulpe mlečnih zuba dece dobijena populacija ćelija koja formira značajan broj CFU-F (4% izolovanih ćelija) i kontinuirano proliferiše tokom 40 dana, bez opadanja vremena potrebnog za udvajanje njihovog broja u kulturi. Ipak, dobijena populacija ćelija je bila heterogena po brzini deoba. Oko 12% ćelija je posle tri dana kultivacije bilo podeljeno manje od četiri puta, dok se ostatak ćelija podelio više od četiri puta. Vijabilitet ćelija je bio u proseku oko 95%, a svega 3% ćelija je bilo pozitivno na β-galaktozidazu. Tokom čitavog vremena kultivacije kapacitet za diferencijaciju u osteoblaste, adipocite i hondrocite se nije menjao. Gotovo sve ćelije su u pasaži četiri eksprimirale CD44, CD73, kolagen tipa I, CD29, CD90 i osteonektin. Deo ćelija je eksprimirao STRO-1, CD146 i CD106. Marker matičnih ćelija hematopoeze nije detektovan u našem sistemu kultivacije. Opseg u kome se kretao procenat ćelija koje pokazuju povišenu aktivnost ALDH je iznosio od 4% do 24% i bio je u negativnoj korelaciji sa vremenom udvajanja ćelija u kulturi. Naši rezultati su pokazali da uticaj NAC-a na izolovane ćelije zavisi od
Galna kiselina u ranijim je istraživanjima potvrđena kao važna antioksidativna sastavnica jestive i ljekovite biljke Peltiphyllum peltatum. U ovome istraživanju ispitan je njen potencijalni zaštitni ...učinak protiv oksidativnog stresa izazvanog natrijevim fluoridom (NaF) u eritrocitima štakora. Oksidativni stres izazvan je davanjem NaF eksperimentalnim životinjama putem vode za piće (1030,675 mg m-3) tijekom jednog tjedna. Galna kiselina životinjama je davana intraperitonealno u dozama 10 mg kg-1 i 20 mg kg-1 na dan. Životinje iz pozitivno kontrolne skupine svaki su dan, osim NaF, intraperitonealno primale vitamin C u dozi 10 mg kg-1. U eritrocitima štakora primjenom TBARS-testa izmjerene su razine lipidne peroksidacije, aktivnosti antioksidativnih enzima (superoksid dismutaze i katalaze) te razina reduciranoga glutationa. Lipidna peroksidacija u štakora izloženih NaF osjetno se povećala (88,8 %) u usporedbi s kontrolnom skupinom (p<0,05). Prethodni tretman galnom kiselinom smanjio je razinu lipidne peroksidacije u eritrocitima, ovisno o dozi. Toksičnost NaF smanjila je aktivnost enzima katalaze i superoksid dismutaze i razine glutationa za 54,4 %, 63,69 %, odnosno 42 % (p<0,001; u usporedbi s kontrolnom skupinom). Dobiveni rezultati upućuju na pozitivne učinke predobrade štakora galnom kiselinom izoliranom iz Peltiphyllum peltatum i vitaminom C na razinu oksidativnog stresa izazvanog natrijevim fluoridom u eritrocitima štakora.
Flail-arm sindrom: prikaz slučaja Bučuk, Mira; Sonnenschein, Ivan; Novak, Petra
Medicina Fluminensis,
12/2014, Volume:
50., Issue:
4
Web Resource
Open access
Cilj: Prikazati rijedak klinički entitet, flail-arm sindrom, varijantu amiotrofične lateralne skleroze. Prikaz slučaja: Prikazujemo šezdesetosmogodišnjeg muškarca s atrofijom proksimalnih mišića ...gornjih ekstremiteta i ramenog pojasa. Miotatski refleksi u gornjim ekstremitetima nisu se izazivali, a u donjim ekstremitetima bili su pojačani. Plantarni je refleks obostrano bio fiziološki. Fascikulacije su bile vidljive u mišićima nadlaktica. Znakova slabosti bulbarnih mišića pacijent nije imao. Tijekom 27 mjeseci nakon nastupa simptoma njegovo je stanje bilo nepromijenjeno. Zaključak: Rezultati pretraga i klinička slika govore u prilog flailarm sindromu.
Radnici u pjeskarenju traper platna izloženi su visokom riziku od silikoze, profesionalne plućne bolesti uzrokovane udisanjem čestica silikatne prašine. Razvoj i progresija silikoze povezani su s ...aktivacijom imunosnog sustava i oksidativnim stresom. Pri aktivaciji imunosnoga sustava, interferon-gama potiče otpuštanje neopterina i enzima indoleamina 2, 3-dioksigenaze (IDO) u različitim vrstama stanica. Određivanje omjera kinurenina i triptofana te koncentracije neopterina pokazale su se učinkovitim metodama praćenja aktivacijskoga statusa IDO-a i staničnog imuniteta. Ovaj rad istražuje uzrokuje li profesionalna izloženost silici promjene u razinama neopterina, degradaciji triptofana i aktivnosti superoksid dismutaze (SOD) i katalaze (CAT), agenata u antioksidativnom obrambenom sustavu. U istraživanju je sudjelovalo 55 muških radnika u pjeskarenju traper platna i 22 zdrava muškarca u kontrolnoj skupini. Srednje vrijednosti razina neopterina i kinurenina, omjera kinurenina i triptofana, te aktivnosti SOD-a bile su više u radnika oboljelih od silikoze nego u kontrolnoj skupini (p<0,05). Razina neopterina i omjer kinurenina i triptofana bile su u pozitivnoj korelaciji (p<0,05). Međutim, korelacija nije uočena između mjerenih vrijednosti i radnog staža. Neke od mjerenih vrijednosti bitno su ovisile o težini patologije. Dobiveni rezultati daju naslutiti da izloženost silici uzrokuje aktivaciju staničnog imunosnog odgovora. Povećane razine neopterina i degradacije triptofana potvrđuju mogućnost njihova korištenja kao pokazatelja staničnog imunosnog odgovora.
kisikovih radikala tako i zbog smanjene aktivnosti obrambenih sustava koji se mogu oduprijeti njihovu djelovanju. Stoga su saznanja o antioksidativnom kapacitetu anestetika koji se primjenjuju prije ...nekoga kirurškog zahvata vrlo važna i od velikog su kliničkog značenja. Sevofl uran i desfl uran su inhalacijski anestetici koji se učestalo rabe u svrhu uvođenja bolesnika u anesteziju. Cilj ovog istraživanja bio je utvrditi razine oksidativnog stresa u različitim tkivima štakora i usporediti razlike u odgovoru tkiva na izlaganje navedenim anesteticima. U tu svrhu razine oksidativnog stresa izmjerili smo u jetri, mozgu, bubrezima i
plućima štakora podijeljenih u tri eksperimentalne skupine. Kontrolna skupina udisala je samo zrak, dok su druge dvije skupine izložene 8 %-tnomu sevofl uranu te 4 %-tnomu desfl uranu tijekom 4 h. Nakon
završetka obrade životinje su žrtvovane i uzimani su im uzorci tkiva za biokemijske analize. Mjerena je razina malondialdehida (MDA), aktivnst enzima superoksid dismutaze (SOD) i glutation peroksidaze
(GSH-Px) te razine bakra i cinka. Izloženost anesteticima izazvala je oksidativni stres u plućima, na što upućuje značajno povišena razina MDA (Mann-Whitney U-test P<0.05) izmjerena u plućnom tkivu štakora obiju izloženih skupina u odnosu na kontrolu. Plućno je tkivo u odnosu na ostala tkiva podložnije štetnim utjecajima reaktivnih kisikovih radikala vjerojatno stoga što je ono prvo izloženo plinovitim anesteticima nakon njihova ulaska u organizam. Razine oksidativnog stresa i antioksidativne aktivnosti koje smo izmjerili u ostalim tkivima bile su različite te su ovisile o primijenjenom anestetiku. Na osnovi dobivenih rezultata možemo zaključiti da bi se zbog različitog odgovora tkiva izbor anestetika trebao provoditi na individualnoj
osnovi.
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