Landa, M*
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High activity of alkaline phosphatase, acid phosphatase and non-specific esterase was demonstrated by histochemical methods in the tegument of scolex of a polycephalic larva of Hydatigera krepkogorski (Schulz et Landa, 1934). The nerve cells and fibres of scolex and bladder exhibited a high activity of alkaline phosphatase and non-specific esterase. Both enzymes were present also in sensory endings. In the bladder, the activity of alkaline and acid phosphatase and non-specific esterase was localized in the tegument and subtegumental cells. High activity of alkaline phosphatase in the tegument of scolex and bladder indicates a high transport of substances in these parts of larva. In the scolex, the activity of alkaline phosphatase was higher than the activity of acid phosphatase, whereas in the bladder tegument and subtegumental cells, the activity of alkaline phosphatase was lower than that of acid phosphatase.
- MeSH
- alkalická fosfatasa analýza MeSH
- Cestoda anatomie a histologie enzymologie MeSH
- esterasy analýza MeSH
- kyselá fosfatasa analýza MeSH
- larva enzymologie MeSH
- močový měchýř enzymologie MeSH
- nervová vlákna enzymologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- alkalická fosfatasa MeSH
- esterasy MeSH
- kyselá fosfatasa MeSH
Approximately 30% of patients with hypertension have hepatic steatosis, and it has recently been proposed that fatty liver be considered a feature of the metabolic syndrome. Obesity, diet, and level of physical activity are likely factors modulating risk for hepatic steatosis, however genetic factors could also influence susceptibility or resistance to fatty liver in hypertensive or normotensive subjects. In genetic studies in spontaneously hypertensive rats (SHRs) and Brown Norway (BN) rats, we discovered that a variant form of sterol regulatory element binding transcription factor 1 (Srebf1 gene, SREBP-1 protein) underlies a quantitative trait locus (QTL) influencing hepatic cholesterol levels in response to a high cholesterol diet. Compared with the BN allele of Srebf1, the SHR allele of Srebf1 includes variants in the promoter and coding regions that are linked to hepatic deficiency of SREBP-1 mRNA and protein, reduced expression of the SREBP-1 target gene stearoyl-CoA desaturase 1, reduced promoter activity for SREBP-1c, and relative protection from dietary induced accumulation of liver cholesterol. Genetic correction of reduced SREBP-1 activity by derivation of congenic and transgenic strains of SHR increased hepatic cholesterol levels, thereby confirming Srebf1 as a QTL influencing hepatic lipid metabolism in the rat. The Srebf1 variant regulating hepatic cholesterol did not appear to affect blood pressure. These findings (1) are consistent with the results of association studies indicating that common polymorphisms affecting SREBP-1 may influence cholesterol synthesis in humans and (2) indicate that variation in Srebf1 may influence risk for hepatic steatosis.
- MeSH
- alely MeSH
- cholesterol metabolismus MeSH
- geneticky modifikovaná zvířata MeSH
- hypertenze komplikace genetika MeSH
- játra metabolismus MeSH
- krevní tlak fyziologie MeSH
- krysa rodu Rattus MeSH
- lokus kvantitativního znaku genetika MeSH
- mutace genetika MeSH
- potkani inbrední BN MeSH
- potkani inbrední SHR MeSH
- protein SREBP1 genetika metabolismus MeSH
- rizikové faktory MeSH
- sekvenční analýza DNA MeSH
- ztučnělá játra komplikace genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- cholesterol MeSH
- protein SREBP1 MeSH
- Srebf1 protein, rat MeSH Prohlížeč
Within the group of higher fungi, edible medicinal mushrooms have a long history of being used as food and in folk medicine. These species contain biologically active substances with many potential beneficial effects on human health. The Pleurotus genus is representative of medicinal mushrooms because Pleurotus ostreatus is one of the most commonly cultivated culinary mushrooms. In our study, we focused on lesser-known species in the genus Pleurotus and measured their antioxidant and anti-inflammatory activity. We prepared extracts of the mushrooms and analyzed them using HPLC-HRMS, GC-MS, and 1H-NMR. Significant differences in biological activities were found among the Pleurotus spp. extracts. A MeOH extract of P. flabellatus was the most active as a radical scavenger with the highest ORAC, while a chloroform extract had significant anti-inflammatory COX-2 activity. The 80% MeOH extract of P. flabellatus contained the highest amounts of ergosterol, ergothioneine, and mannitol. The 80% MeOH extract of P. ostreatus Florida was the most active in the NF-κB inhibition assay and had the highest content of β-glucans (43.3% by dry weight). Given the antioxidant and anti-inflammatory properties of P. flabellatus, the potential therapeutic usefulness of this species is worth evaluating through in-depth investigations and confirmation by clinical trials.
To identify renally expressed genes that influence risk for hypertension, we integrated expression quantitative trait locus (QTL) analysis of the kidney with genome-wide correlation analysis of renal expression profiles and blood pressure in recombinant inbred strains derived from the spontaneously hypertensive rat (SHR). This strategy, together with renal transplantation studies in SHR progenitor, transgenic and congenic strains, identified deficient renal expression of Cd36 encoding fatty acid translocase as a genetically determined risk factor for spontaneous hypertension.
- MeSH
- antigeny CD36 genetika MeSH
- hypertenze genetika MeSH
- krysa rodu Rattus MeSH
- ledviny metabolismus MeSH
- lokus kvantitativního znaku MeSH
- potkani inbrední SHR MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- antigeny CD36 MeSH
Behavioral sensitization is a phenomenon occurring after repeated administration of various psychotropic substances and it is characterized by gradually increasing response to the particular drug. It has been described for majority of addictive substances including amphetamines. It is considered to reinstate drug-seeking behaviour and plays important role in the processes associated with drug abuse and addiction. There are published reports, particularly on preclinical level, that N-acetylcysteine (NAC) may affect addictive properties of different classes of drugs (e.g., cocaine, heroin, alcohol, cannabinoids, nicotine). Since the lack of information on possible effects of NAC on amphetamine derivatives we decided to test possible influence of this substance on behavioral sensitization to methamphetamine (MET) in the mouse open field test. Our results have shown a decreased acute stimulatory effect of MET caused by NAC and moreover, there was a non-significant trend of attenuated development of behavioral sensitization to MET after simultaneous long-term administration of MET and NAC. This suppression of MET stimulatory effects therefore suggested on the preclinical level possible promising efficacy of NAC on addictive properties associated with MET similarly as it was demonstrated by other authors in association with cocaine or heroin. Key words: N-acetylcysteine, Methamphetamine, Behavioral sensitization.
- MeSH
- acetylcystein * farmakologie aplikace a dávkování MeSH
- chování zvířat * účinky léků MeSH
- methamfetamin * farmakologie aplikace a dávkování MeSH
- myši MeSH
- stimulanty centrálního nervového systému * farmakologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- acetylcystein * MeSH
- methamfetamin * MeSH
- stimulanty centrálního nervového systému * MeSH
WAP is being recognized as the principal milk protein expressed in pregnant or lactating females of several mammalian species. Recently, it has been shown that the 6.3-kb 5' untranslated region of the rWAP gene is able to control, and almost completely restrict, the expression of the transgene into the mammary gland of the transgenic animal. We cloned the genomic fragment carrying the rWAP gene locus from the rabbit phage genomic library and used the 8.5-kb long 5' untranslated part of the rWAP gene to target the expression of hEPO, cloned from the human phage genomic library, into the mammary gland of the mouse. The vectors, carrying either the hEPO gene or the rWAP-hEPO hybrid gene, were injected into the mouse ova, and 12 transgenic animals were identified by PCR and Southern blot from the progeny of 168 tested littermates. Transgenic mice were viable, fertile and displayed a normal development. Recombinant human erythropoietin was produced in the milk of a transgenic mouse female at a secretion level of 5.3 mIU/ml, as detected by ELISA. Despite the low production of the transgenic glycoprotein in the milk we demonstrate that the hybrid gene can be expressed in the mammary gland of the host animal. Thus, WAP-based recombinant vectors, with additional optimizing modifications, can be useful for production of therapeutic proteins in the transgenic mammals.
- MeSH
- erythropoetin krev genetika MeSH
- exprese genu MeSH
- lidé MeSH
- mikroinjekce MeSH
- mléčné bílkoviny genetika MeSH
- mléčné žlázy zvířat fyziologie MeSH
- myši transgenní MeSH
- myši MeSH
- polymerázová řetězová reakce MeSH
- promotorové oblasti (genetika) genetika MeSH
- transgeny MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- erythropoetin MeSH
- mléčné bílkoviny MeSH
- whey acidic proteins MeSH Prohlížeč
Bovine in vitro matured oocytes were parthenogenetically activated by a single pulse of direct current. Suppression of second polar body extrusion by 60 V DC-pulse followed by cytochalasin B treatment or 240 V DC-pulse were used to diploidize parthenogenones. Analysis of the cytological events 20 h postactivation clearly documented high efficiency of both diploidization techniques. All oocytes exposed to a single 60 V DC-pulse and then to cytochalasin B for 3 to 6 h were activated and 70% to 88% possessed two pronuclei. Exposure to 240-V DC-pulse for 10 and 20 microseconds activated all oocytes, 72% and 62% formed two pronuclei, but a high incidence (22% and 30%) of degenerated oocytes was observed. Prolonged in vitro culture of oocytes after parthenogenetic activation and diploidization treatment showed that formed pronuclei were able to fuse and single prometaphase to telophase mitotic cleavage figures developed in all oocytes fixed 28 h after activation, except one cytochalasin B-treated oocyte with two prometaphase sets of chromosomes.
- MeSH
- cytochalasin B farmakologie MeSH
- diploidie MeSH
- elektrická stimulace MeSH
- mitóza účinky léků MeSH
- oocyty cytologie účinky léků MeSH
- partenogeneze účinky léků genetika MeSH
- skot MeSH
- techniky in vitro MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- cytochalasin B MeSH
Tubal cumulus-free oocytes recovered from C57BL/6J and BALB/c mice 16 to 24 h after hCG injection were parthenogenetically activated by a single DC-pulse of different voltage. The efficiency of activation was assessed 2, 4 and 24 h after electric stimulation according to the proportion of oocytes with second polar bodies, presence of pronuclei and development of parthenogenetic 2-cell embryos. The best results, 99% and 100% of oocytes with second polar body (2 h) or with second polar body and one pronucleus (4 h), were recorded when oocytes of both strains were activated by a single 60 V DC-pulse (30 microseconds) 20 h after hCG injection. Aging of oocytes or higher voltage of DC-pulse (120 V and 240 V) resulted in both strains in an increase of abnormal oocytes and oocytes with two pronuclei. While a large proportion of activated C57BL/6J oocytes were totally fragmented within 24 h and only 23% reached 2-cell stage, 78% of the activated BALB/c oocytes developed to parthenogenetic 2-cell embryos. It was shown that pronuclear oocytes and parthenogenetic 2-cell embryos for preparation of different types of cytoplasts could be effectively prepared by electric stimulation of tubal oocytes.
- MeSH
- buněčná diferenciace MeSH
- choriogonadotropin aplikace a dávkování MeSH
- elektrická stimulace MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- oocyty cytologie MeSH
- partenogeneze * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- choriogonadotropin MeSH
Production of recombinant human erythropoietin (rhEPO) for therapeutic purposes relies on its expression in selected clones of transfected mammalian cells. Alternatively, this glycoprotein can be produced by targeted secretion into the body fluid of transgenic mammals. Here, we report on the generation of a transgenic rabbits producing rhEPO in the lactating mammary gland. Transgenic individuals are viable, fertile and transmit the rhEPO gene to the offspring. Northern blot data indicated that the expression of the transgene in the mammary gland is controlled by whey acidic protien (WAP) regulatory sequences during the period of lactation. While the hybridization with total RNA revealed the expression only in the lactating mammary gland, the highly sensitive combinatory approach using RT-PCR/hybridization technique detected a minor ectopic expression. The level of rhEPO secretion in the founder female, measured in the period of lactation, varied in the range of 60-178 and 60-162 mIU/ml in the milk and blood plasma, respectively. Biological activity of the milk rhEPO was confirmed by a standard [3H]-thymidine incorporation test. Thus, we describe the model of a rhEPO-transgenic rabbit, valuable for studies of rhEPO glycosylation and function, which can be useful for the development of transgenic approaches designed for the preparation of recombinant proteins by alternative biopharmaceutical production.
- MeSH
- erythropoetin metabolismus MeSH
- geneticky modifikovaná zvířata genetika MeSH
- izotopové značení MeSH
- králíci genetika MeSH
- laktace metabolismus MeSH
- lidé MeSH
- mléčné bílkoviny genetika MeSH
- mléčné žlázy zvířat metabolismus MeSH
- mléko metabolismus MeSH
- molekulární sekvence - údaje MeSH
- regulace genové exprese genetika MeSH
- rekombinantní proteiny MeSH
- sekvence nukleotidů MeSH
- thymidin metabolismus MeSH
- zvířata MeSH
- Check Tag
- králíci genetika MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- erythropoetin MeSH
- mléčné bílkoviny MeSH
- rekombinantní proteiny MeSH
- thymidin MeSH
- whey acidic proteins MeSH Prohlížeč
Transgenic mice were produced from DNA-injected embryos stored for 2 to 30 days in liquid nitrogen. Of the 500 zygotes collected from (C57BL/6 x CBA)F1 mice, 363 (73%) survived DNA injection into pronuclei and 246 (82%) morphologically normal 4- and 8-cell embryos were flushed from temporary recipients 48 h later. Of the 200 DNA-injected 8-cell embryos cryopreserved by vitrification in microdrops, 194 (97%) were recovered and 188 (94%) embryos were intact one hour after thawing. Of the 50 DNA-injected and frozen/thawed embryos, 48 (96%) developed to morulae or blastocysts within 30 h of in vitro culture. Transfer of 100 DNA-injected and cryopreserved 8-cell embryos into 20 day-1 recipients resulted in 47 young born. Two mice were transgenic.
- MeSH
- DNA genetika MeSH
- embryo savčí fyziologie MeSH
- erythropoetin genetika MeSH
- exprese genu MeSH
- kryoprezervace * MeSH
- mikroinjekce MeSH
- míra přežití MeSH
- myši inbrední C57BL MeSH
- myši inbrední CBA MeSH
- myši transgenní genetika MeSH
- myši MeSH
- přenos embrya MeSH
- transfekce * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA MeSH
- erythropoetin MeSH