Comparison of Manual and Automatic (MagNA Pure) Isolation Methods of Total RNA from Adipose Tissue

Aim Comparison of manual and automatic (MagNA Pure) isolation methods of total RNA from adipose tissue with respect to its quality and recovery factor. Material 120 human subcutaneous adipose tissue samples (about 100 mg/sample) were collected from patients during surgical operations. The tissue sample was stabilized in RNAlater (QIAGEN GmbH, Germany). Methods Total RNA was extracted by the following kits: Rneasy Protect Mini, Rneasy Lipid Tissue (QIAGEN GmbH, Germany) and MagNA Pure Compact RNA Isolation (Tissue) for MagNA Pure Compact Instrument (Roche Diagnostics GmbH, Germany). Results The average RNA yields with Rneasy Lipid Tissue kits were about two-fold higher in comparison with the Rneasy Protect Mini kit. When the MagNA Pure Compact System was used, RNA yields from the same sample were more uniform compared with manual systems. It was also more convenient and less time-consuming than the manual approach. No DNA contamination of total RNA samples was detected except for samples isolated by Rneasy Protect Mini Kit. Conclusion Rneasy Lipid Tissue Kit and MagNA Pure Compact RNA Isolation Kit (Tissue) provide RNA samples of high quantity, purity and PCR amplificability. RNA samples are suitable for further processing using methods of molecular biology.     

Sexual dimorphism in the size of nuclei with dioecious plants

Measurements of the size of the nuclei of dioecious plants showed that the nuclei of male and female plants differ in agreement with the larger quantity of chromatin. The male. plants ofRumex acetosella andMelandrium album had larger nuclei, their Y chromosome being larger than the X chromosome, the same is true forRumex acetosa where the Y chromosome is smaller but there are two in the set.Ginkgo biloba had larger female nuclei because the Y chromosome was smaller than the X. The curves obtained by grouping all the nuclei of both sexes never had two peaks with regard to the small differences between the classes of maximum frequency.     

Brevibacterium divaricatum: Influence of nutritional conditions on the wall composition and release of uncross-linked peptidoglycan chains into medium

Purified cell walls, originating from penicillin-treated (3 g/ml, 1 h) and-untreated Brevibacterium divaricatum cells grown on complex (CM) and glucose minimal medium with (MM) or without (Ca-free MM) calcium carbonate, were isolated by two procedures. Electron micrographs and chemical analysis revealed no differences between identically isolated walls with respect to the presence or absence of either penicillin or calcium carbonate in the glucose growth medium. On the contrary, the appearance and peptidoglycan content of the walls was greatly dependent on the procedure used for their isolation and the walls isolated from the cells grown on complex medium contained more materials other than peptidoglycan. It was shown that the presence of calcium carbonate in the glucose minimal medium was essential for accumulation of large amounts of peptidoglycan chains into the medium. Penicillin-induced interruption of cell wall synthesis was prerequisite for manifestation of the calcium carbonate stimulating effect.Abbreviations CM complex medium - MM chemically defined minimal medium based on glucose and containing calcium carbonate - Ca-free MM MM modified only by the omission of calcium carbonate - ET-walls Enzyme treated walls - FPR-walls French press-ruptured walls     

Constrained sessile drop as a new configuration to measure low surface tension in lung surfactant systems.

Existing methodology for surface tension measurements based on drop shapes suffers from the shortcoming that it is not capable to function at very low surface tension if the liquid dispersion is opaque, such as therapeutic lung surfactants at clinically relevant concentrations. The novel configuration proposed here removes the two big restrictions, i.e., the film leakage problem that is encountered with such methods as the pulsating bubble surfactometer as well as the pendant drop arrangement, and the problem of the opaqueness of the liquid, as in the original captive bubble arrangement. A sharp knife edge is the key design feature in the constrained sessile drop that avoids film leakage at low surface tension. The use of the constrained sessile drop configuration in conjunction with axisymmetric drop shape analysis to measure surface tension allows complete automation of the setup. Dynamic studies with lung surfactant can be performed readily by changing the volume of a sessile drop, and thus the surface area, by means of a motor-driven syringe. To illustrate the validity of using this configuration, experiments were performed using an exogenous lung surfactant preparation, bovine lipid extract surfactant (BLES) at 5.0 mg/ml. A comparison of results obtained for BLES at low concentration between the constrained sessile drop and captive bubble arrangement shows excellent agreement between the two approaches. When the surface area of the BLES film (0.5 mg/ml) was compressed by about the same amount in both systems, the minimum surface tensions attained were identical within the 95% confidence limits.     

Beitrag zur Kenntnis desAlnetum incanae in der Tschechischen Sozialistischen Republik (ČSR)

Gegenstand dieses Beitrages ist die syntaxonomisch-synökologische Charakteristik desAlnetum incanae Lüdi 1921 in den Tschechischen Ländern. Bestände dieser montanen Auenwaldassoziation begleiten hier Flussgewässer in Gebirgstälern und am Hochgebirgsrand, meist in den Lagen von 650–900 mü. NN. DasAlnetum incanae wird in zwei Subassoziationen geteilt. Das feuchteliebendeAlnetum incanae calthetosum Knapp exNeuhäuslová-Novotná subass. nova ist an Gleyböden (Nassgleye, sauerstoffreiche Gleye) gebunden, dasAlnetum incanae typicum Oberd. 1949 (mit zwei Varianten)bestockt Paternia-Böden. DieAegopodium podagraria-Variante der typischen Subassoziation bevorzugt pedogenetisch mehr entwickelte Böden mit tiefem Grund-wasser, meist in der Kontaktzone zu Klimaxgesellschaften.     

Beitrag zur Kenntnis der Waldgesellschaften der Kleinen Karpaten,Slowakei 2. Standortsverhältnisse

In diesem Beitrag werden die Standortsverhältnisse der kleinkarpatischen Laubwaldgesellschaften (Carici pilosae-Carpinetum, sekundärerPrimula-Eichen-Hainbuchenwald,Festuco heterophyllae-Quercetum, Stellario-Alnetum, Carici remotae-Fraxinetum, Dentario bulbiferae-Fagetum, Cephalanthero-Fagetum, Phyllitidi-Aceretum, Luzulo-Fagetum, Eu-Quercion pubescentis, Luzulo albidae-Quercetum) behandelt. Besonders wurden die Beziehungen der genannten Gesellschaften zu den physikalischen und chemischen Bodeneigenschaften und Bodentypen sowie ihre Höhenverbreitung verfolgt. Ein zusammenfassender Standortsvergleich dieser Gesellschaften mit einem Bestimmungsschlüssel nach Standortsverhältnisschen ist beigefügt.