gradient analysis
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Cieľ: Lamina cribriformis je hranicou 2 tlakov: vnútroočného tlaku (VOT) a intrakraniálneho tlaku (IKT). Rozdiel medzi VOT a IKT sa označuje ako translaminárny gradient (TLG). Cieľom bolo sledovať glakomatóznu progresiu (zorné pole, fundus vyšetrenie, HRT vyšetrenia) bez / s lokálnou antiglaukomatóznou terapiou) vo vzťahu k TLG. Pacienti a metodika: Hodnotenie významu TLG bolo skúmané v dvoch skupinách. I. skupina 57 pacientov s diagnostikovaným a liečeným primárnym glaukómom s otvoreným uhlom (PGOU), 10 pacientov s okulárnou hypertenziou (OH), 7 pacientov s nízkotenzným glaukómom (NTG) a 75 zdravých bez glaukómu. Hodnotenia TLG boli realizované jednorázovo a retrospektívne. V II. skupine boli prospektívne sledovaní 14 pacienti s OH a 24 pacientov s novozisteným PGOU bez nastavenej terapie. Vyšetrenia boli realizované 4x s odstupom 10–11 mesiacov. Všetky vyšetrenia zahrňovali základné očné vyšetrenia, ORA tonometriu, HRT vyšetrenie, gonioskopiu, farebný Doppler sonografiu ciev oka a očnice. Venózny pulzačný tlak (VPT) bol zaznamenaný oftalmodynamometrom Meditron (D-ODM). V prípadoch spontánnej venóznej pulzácie Vena centralis retinae, bol VPT považovaný za tlak rovnaký ako VOT Pre výpočet TLG bol použitý vzorec podľa Querfurtha IKT = 0,29 + 0,74 (VOT / PI (AO )). [PI(AO) – index pulzatility z arteria ophthalmica (AO)]. Výsledky: I. skupina: TLG bol v kontrolnej skupine bez glaukómu: 12,2 ? 2,0 torr. V skupine NTG: 9,0 ? 1,70 torr. PGOU: 11,1 ? 1,91 torr. OH: 12,6 ? 0.85 torr. Samotný IKT neprejavuje signifikantný vzťah k prítomnosti glaukómu, okulárnej hypertenzie. II. skupina: Pacienti s OH (14 pacientov) mali TLG v 12 prípadoch 3,8 ? 1,2 torr. 2 pacienti (OH) mali TLG 10 torr. a 15 torr. U jedného z nich (TLG = 15 torr.) bola po 4 rokoch zaznamenané glaukomatózna progresia. V skupine PGOU pred liečbou bol u všetkých pacientov bol TLG 15,0 ? 4,8 torr. Po nastavení lokálnej antiglaukomatóznej terapie a úprave VOT, sa u 20 pacientov znížil TLG na 3,6 ? 1,3 torr. Záver: TLG preukázal signifikantný vzťah k progresii glaukómu. Riziko glaukomatózneho poškodenia stúpa priamo úmerne so zvyšujúcou sa hodnotou translaminárneho gradientu. Translaminárny gradient slúži pre upresnenie tzv. „cieľového“ vnútroočného tlaku. Hodnotenie TLG má význam pri okulárnych poškodeniach (okulárna hypertenzia, glaukóm, cievne oklúzie, neuropatie zrakového nervu), intrakraniálnych procesoch, orbitopatiách, pre výber vhodného antiglaukomatika.
Objective: The cribriform plate is a threshold of the intraocular pressure (VOT) and of the intracranial pressure (IKT). The difference between the VOT and IKT is referred to as translaminar gradient (TLG). The goal was to evaluate the Glaucoma progression (visual field, fundus examination, HRT) with / without topical anti-glaucomatous therapy) in relation to the TLG. Patients and methods: the significance of TLG has been studied in two groups. I. Group: 57 patients diagnosed and treatment of Primary Open-Angle Glaucoma (PGOU), 10 patients with Ocular hypertension (OH), 7 patients with Normal-Tension Glaucoma (NTG), and 75 healthy without glaucoma. The examinations of TLG were carried out once and retrospectively. In II. group there were prospectively studied 14 patients with OH and 24 patients with newly detected PGOU without local therapy. The examinations were performed 4 times at intervals of 10 to 11 months. All tests included a basic eye examination, ORA tonometry, HRT examination, gonioscopy, Color Doppler sonography of blood vessels of the eye and orbit. Venous pulsation pressure (VPT) has been recorded by the Ophthalmodynamometer Meditron (D-ODM). In case of spontaneous retinal venous pulsation, VPT was considered as the same pressure as the VOT. The TLG was calculated with formula of Querfurth: ICT = 0.29 + 0.74 (VOT / PI (AO)). [PI(AO) – Pulsatility index of the Ophthalmic artery (AO)]. Results: I. group: TLG was in the control group without Glaucoma: 12.2 ? 2.0 torr. The NTG group: 9.0 ? 1.70 mm Hg. PGOU: 11.1 ? 1.91 mm Hg. OH: 12.6 ? 0.85 mm Hg. IKT alone does not show a significant relationship to the presence of glaucoma, ocular hypertension. II. Group: The average TLG in Ocular Hypertension (14 patients) has been 3.8 ? 1.2 torr. 2 patients (OH) had TLG 10 torr. and 15 torr. After 4 years in one of them (TLG = 15 torr.) there was recorded Glaucoma progression. In the PGOU group before antiglaucoma therapy, TLG was 15.0 ? 4.8 torr for all patients. After setting up local anti-glaucoma therapy and decreasing VOT, the TLG in 20 patients reduced to 3.6 ? 1.3 mm Hg. Conclusion: TLG showed a significant relationship to the Glaucoma progression. The risk of glaucomatous damage increases proportionally with increasing Translaminar gradient. Translaminar gradient can be use to refine the so-called. “Target VOT”. TLG has a role in ocular damage (ocular hypertension, glaucoma, vascular occlusion, optic neuropathy), intracranial damage, orbitopathy, selection of appropriate antiglaucomatous therapy.
- Klíčová slova
- translaminární gradient,
- MeSH
- glaukom s otevřeným úhlem patofyziologie terapie MeSH
- glaukom * patofyziologie terapie MeSH
- intrakraniální tlak MeSH
- kohortové studie MeSH
- lidé MeSH
- nitrooční tlak MeSH
- oční hypertenze MeSH
- oftalmodynamometrie MeSH
- progrese nemoci MeSH
- zrakové testy MeSH
- Check Tag
- lidé MeSH
Velocity separation of translation complexes in linear sucrose gradients is the ultimate method for both analysis of the overall fitness of protein synthesis as well as for detailed investigation of physiological roles played by individual factors of the translational machinery. Polysome profile analysis is a frequently performed task in translational control research that not only enables direct monitoring of the efficiency of translation but can easily be extended with a wide range of downstream applications such as Northern and Western blotting, genome-wide microarray analysis or qRT-PCR. This chapter provides a basic overview of the polysome profile analysis technique and the RNA isolation procedure from sucrose gradients. We also discuss possible experimental pitfalls of data normalization, describe main alternatives of the basic protocol and outline a novel application of denaturing RNA electrophoresis in several steps of polysome profile analysis.
- MeSH
- centrifugace - gradient hustoty metody MeSH
- elektroforéza metody MeSH
- interpretace statistických dat MeSH
- kvasinky MeSH
- polyribozomy chemie MeSH
- proteosyntéza genetika MeSH
- regulace genové exprese genetika MeSH
- RNA izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Objective: The cribriform plate is a threshold of the intraocular pressure (VOT) and of the intracranial pressure (IKT). The difference between the VOT and IKT is referred to as translaminar gradient (TLG). The goal was to evaluate the Glaucoma progression (visual field, fundus examination, HRT) with / without topical anti-glaucomatous therapy) in relation to the TLG. Patients and methods: the significance of TLG has been studied in two groups. I. Group: 57 patients diagnosed and treatment of Primary Open-Angle Glaucoma (PGOU), 10 patients with Ocular hypertension (OH), 7 patients with Normal-Tension Glaucoma (NTG), and 75 healthy without glaucoma. The examinations of TLG were carried out once and retrospectively. In II. group there were prospectively studied 14 patients with OH and 24 patients with newly detected PGOU without local therapy. The examinations were performed 4 times at intervals of 10 to 11 months. All tests included a basic eye examination, ORA tonometry, HRT examination, gonioscopy, Color Doppler sonography of blood vessels of the eye and orbit. Venous pulsation pressure (VPT) has been recorded by the Ophthalmodynamometer Meditron (D-ODM). In case of pontaneous retinal venous pulsation, VPT was considered as the same pressure as the VOT. The TLG was calculated with formula of Querfurth: ICT = 0.29 + 0.74 (VOT / PI (AO)). [PI(AO) – Pulsatility index of the Ophthalmic artery (AO)]. Results: I. group: TLG was in the control group without Glaucoma: 12.2 ± 2.0 torr. The NTG group: 9.0 ± 1.70 mm Hg. PGOU: 11.1 ± 1.91 mm Hg. OH: 12.6 ± 0.85 mm Hg. IKT alone does not show a significant relationship to the presence of glaucoma, ocular hypertension. II. Group: The average TLG in Ocular Hypertension (14 patients) has been 3.8 ± 1.2 torr. 2 patients (OH) had TLG 10 torr. and 15 torr. After 4 years in one of them (TLG = 15 torr.) there was recorded Glaucoma progression. In the PGOU group before antiglaucoma therapy, TLG was 15.0 ± 4.8 torr for all patients. After setting up local anti-glaucoma therapy and decreasing VOT, the TLG in 20 patients reduced to 3.6 ± 1.3 mm Hg. Conclusion: TLG showed a significant relationship to the Glaucoma progression. The risk of glaucomatous damage increases proportionally with increasing Translaminar gradient. Translaminar gradient can be use to refine the so-called. “Target VOT”. TLG has a role in ocular damage (ocular hypertension, glaucoma, vascular occlusion, optic neuropathy), intracranial damage, orbitopathy, selection of appropriate antiglaucomatous therapy.
- Klíčová slova
- translaminární gradient,
- MeSH
- glaukom s otevřeným úhlem patofyziologie terapie MeSH
- glaukom * patofyziologie terapie MeSH
- intrakraniální tlak MeSH
- kohortové studie MeSH
- lidé MeSH
- nitrooční tlak MeSH
- oční hypertenze MeSH
- oftalmodynamometrie MeSH
- progrese nemoci MeSH
- zrakové testy MeSH
- Check Tag
- lidé MeSH
- MeSH
- lidé MeSH
- mutační analýza DNA metody MeSH
- ženská infertilita diagnóza genetika MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
Soils play an important role in the ecosystem of karstic landscapes both as a buffer zone and as a source of acidity to belowground water. Although the microbiota of karstic soils is known to have a great effect on karstification processes, the activity and composition of these communities are largely unknown. This study gives a comparative analysis of soil microbial profiles from different parts of a doline located at Aggtelek, Hungary. The aim was to reveal the relationships between the vegetation type and genetic fingerprints and substrate utilisation (multi-SIR) profiles of the soil microbiota. Soil samples were collected in early and late springs along a transect in a doline covered with different types of vegetation. Genetic fingerprints of bacterial communities were examined by denaturing gradient gel electrophoresis (DGGE) based on the 16S rRNA gene, along with multi-SIR profiles of the microbial communities measured by the MicroResp method using 15 different carbon sources. Genetic fingerprinting indicated that vegetation cover had a strong effect on the composition of soil bacterial communities. Procrustean analysis showed only a weak connection between DGGE and multi-SIR profiles, probably due to the high functional redundancy of the communities. Seasonality had a significant effect on substrate usage, which can be an important factor to consider in future studies.
- MeSH
- Bacteria klasifikace genetika metabolismus MeSH
- denaturační gradientová gelová elektroforéza MeSH
- geologické jevy MeSH
- mikrobiota fyziologie MeSH
- půda chemie MeSH
- půdní mikrobiologie * MeSH
- RNA ribozomální 16S genetika MeSH
- roční období MeSH
- shluková analýza MeSH
- uhlík metabolismus MeSH
- veřejné parky * MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Maďarsko MeSH
In many fish species, sperm cryopreservation has deleterious effects and leads to a significant decrease in spermatozoa viability. However, the effect of cryopreservation on sperm cells that survive this process and are still viable is not fully understood. The objective of this study was to compare the viability and proteomes of fresh and cryopreserved sterlet (Acipenser ruthenus) sperm samples before and after live-dead cell separation using Percoll density gradient centrifugation. Both fresh and cryopreserved sperm samples were divided into two groups (with or without application of Percoll separation). At each step of the experiment, sperm quality was evaluated by video microscopy combined with integrated computer-assisted sperm analysis software and flow cytometry for live-dead sperm viability analysis. Sperm motility and the percentage of live cells were reduced in the cryopreserved group compared to the fresh group from 89% to 33% for percentage of motility and from 96% to 70% for live cells. Straight line velocity and linearity of track were significantly lower in cryopreserved samples than in those separated by Percoll before and after cryopreservation. However, the percentages of motile and live spermatozoa were higher than 90% in samples subjected to Percoll separation. Proteomic analysis of spermatozoa by two-dimensional differences in-gel electrophoresis coupled with matrix-assisted laser-desorption/ionization time-of-flight/time-of-flight mass spectrometry revealed that 20 protein spot abundances underwent significant changes in cryopreserved samples compared to fresh ones. However, only one protein spot was significantly altered when samples before and after cryopreservation followed by Percoll separation were compared. Thus, the results of this study show that cryopreservation leads to minimal proteomic changes in the spermatozoa population, retaining high motility and viability parameters. The results also suggest that global differences in protein profiles between unselected fresh and cryopreserved samples are mainly due to protein loss or changes in the lethal and sublethal damaged cell subpopulations.
- MeSH
- centrifugace - gradient hustoty metody MeSH
- kryoprezervace metody MeSH
- motilita spermií fyziologie MeSH
- oxid křemičitý chemie MeSH
- povidon chemie MeSH
- proteomika MeSH
- ryby fyziologie MeSH
- spermie fyziologie MeSH
- uchování spermatu metody MeSH
- viabilita buněk fyziologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
More than one 80S monosome can translate an mRNA molecule at a time producing polysomes. The most widely used method to separate 40S and 60S ribosomal subunits from 80S monosomes and polysomes is a high-velocity centrifugation of whole cell extracts in linear sucrose gradients. This polysome profile analysis technique has been routinely used to monitor translational fitness of cells under a variety of physiological conditions, to investigate functions of initiation factors involved in translation, to reveal defects in ribosome biogenesis, to determine roles of 5' UTR structures on mRNA translatability, and more recently for examination of miRNA-mediated translational repression (see an application of this protocol on Polysome analysis for determining mRNA and ribosome association in Saccharomyces cerevisiae).
