Calculating reaction rate constants and estimating the efficacy of selective enzyme inhibitors

The dynamic steady state of a pair of forward and backward enzymatic reactions is dependent on the balance between the enzymes catalyzing the reactions. By selectively inhibiting one or more of the enzymes involved, this balance is shifted into a new steady state, making it possible to calculate the reaction rate constants after measurement of the reactants. Ideally, the inhibitors should completely eliminate either reaction, but this is often not the case. Here we present and discuss a method for calculating the reaction rate constants and, thus, for evaluating the efficacy of one or more inhibitors when introduced to a forward-backward pair of enzymatic reactions.     

Combined effects of Cu,Zn, and Hg on the increase in length of Ascophyllum nodosum (L.) Le Jolis

The increase in length of apices of Ascophyllum nodosum (L.) Le Jolis was measured in paired combinations of Cu, Hg, and Zn.Significant antagonistic effects appeared with exposure to Cu + Zn after 3 days (P < 0.001), and with Hg and Zn after 4 days (P < 0.01). When Cu and Hg were combined, only a weak antagonistic effect was indicated after 7 days of exposure. It is supposed that the observed detoxificating effect of Zn to Cu and Hg is partly related to the relative number of ions present.     

Measurement error and confidence intervals for ROC curves

Measurement error in a continuous test variable may bias estimates of the summary properties of receiver operating characteristics (ROC) curves. Typically, unbiased measurement error will reduce the diagnostic potential of a continuous test variable. This paper explores the effects of possibly heterogenous measurement error on estimated ROC curves for binormal test variables. Corrected estimators for specific points on the curve are derived under the assumption of known or estimated measurement variances for individual test results. These estimators and associated confidence intervals do not depend on normal assumptions for the distribution of the measurement error and are shown to be approximately unbiased for moderate size samples in a simulation study. An application from a study of emerging imaging modalities in breast cancer is used to demonstrate the new techniques.     

The effect of local nitric oxide synthase inhibition on the diameter of pulpal arteriole in dental bond material-induced vasodilation in rat

Local application of dental bond materials can cause pulpal vasodilation and hyperemia. Such local hemodynamic changes may be mediated by alterations in the levels of locally generated nitric oxide (NO). In different species systemic administration of NO synthase inhibitors leads to a decrease in pulpal blood flow. In contrast, the local administration of these inhibitors has not been tested yet. Therefore, the effect of locally blocked NO synthase on the internal diameter of rat pulpal arterioles under basal conditions and immediately after dental bond material application was studied by using vitalmicroscopic technique. The NO synthase blocker (L-NAME) was locally administered on a thinned dentine layer of the left lower incisor. L-NAME reduced the diameter of the pulpal arteriole both in basal and after bond material-induced hyperaemic conditions. These data suggest that the local formation of NO may have a significant role in the acute vasodilation induced by bond material application and also in maintenance of basal pulpal arteriolar tone.     

Vegetative reproduction and chemical control with post-emergent herbicides of field bindweed (Convolvulus arvensis L.)

It is clearly seen from data that roots of Convolvulus arvensis L. have more and less intensive regenerative period during growing season. The more intensive period is in autumn, because in that time roots culminate nutrients, carbohydrate as starch and sugar. The less intensive regenerative or shoot-growing period is in spring, called "late spring bud dormancy". Experiments were conducted to get more information and further details about the regenerative capacity of roots close to and far from the collar of Convolvulus arvensis L. Root segments closer to collar have an intensive regenerative capacity than those ones further to collar. By data of Bakke et al. (1939) is well known, roots exhumed from deep soil layers are able to create shoots with low intensity. So finally we can exclaim that regenerative capacity is decreasing further to collar. Using mechanical weed control it is sufficient to till the upper layer of soil, but many times. Chemical treatments are most effective in the integrated weed control. It is clearly seen that auxin-type herbicide such as 2,4-D, fluroxipir, MCPA. dicamba give the best result. They gave 95% weed control effect used them separately or in combination with other herbicides. Combination of Banvel 480 S (dicamba) and Logran 75 WG (triasulfuron) introduced 95% weed control effect. Only one time got absolutely 100% weed control effect, in the case of Glyphosate active substance. Caused total plant destruction. Excellent result was given with the application of Pledge 50WP (flumioxazin). Herbicides mentioned above are absolutely allowed to take an important and significant part in chemical plant protection against Convolvulus arvensis L. Other herbicides like Granstar 75DF (tribenuron-methyl), Basis 75DF (rimsulfuron + tifensulfuron-methyl) and Huszár (jodosulfuron-methyl-sodium + mefenpir-diethyl) are not so effective against Convolvulus arvensis L., as compared to the previous ones.     

KLN-5: a safe monocationic lipophosphoramide to transfect efficiently haematopoietic cell lines and human CD34+ cells

The safe and efficient delivery of nucleic acids into haematopoietic stem cells (HSCs) has a wide range of therapeutic applications. Although viruses are being used in most clinical trials owing to their high transfection efficacy, recent results highlight many concerns about their use. Synthetic transfection reagents, in contrast, have the advantage of being safe and easy to manage while their low transfection efficiency remains a hurdle that needs to be addressed before they can be widely used. Using information on transfection mechanisms, a new family of monocationic lipids called lipophosphoramides was synthesized. Their efficiency to transfer genes into haematopoietic cell lines (K562, Jurkat and Daudi) and CD34+ cells was assessed. In this study, we report that one of these new compounds, KLN-5, leads to more efficient transfection activity than one of our previously most efficient reagents (EG-308) and the commercially available monocationic lipids (DC-CHOL and DOTAP/DOPE) (P<0.05). In addition, only a slight toxicity related to the chemical structure of the new compounds is observed. Moreover, we show that KLN-5 can successfully carry the transgene into haematopoietic progenitor cells (CD34+). These results demonstrate that synthetic transfection reagents represent a viable alternative to viruses and could have potential practical utility in a number of applications.     

Changes in the root-associated fungal communities along a primary succession gradient analysed by 454 pyrosequencing

We investigated changes in the root-associated fungal communities associated with the ectomycorrhizal herb Bistorta vivipara along a primary succession gradient using 454 amplicon sequencing. Our main objective was to assess the degree of variation in fungal richness and community composition as vegetation cover increases along the chronosequence. Sixty root systems of B. vivipara were sampled in vegetation zones delimited by dated moraines in front of a retreating glacier in Norway. We extracted DNA from rinsed root systems, amplified the ITS1 region using fungal-specific primers and analysed the amplicons using 454 sequencing. Between 437 and 5063 sequences were obtained from each root system. Clustering analyses using a 98.5% sequence similarity cut-off yielded a total of 470 operational taxonomic units (OTUs), excluding singletons. Between eight and 41 fungal OTUs were detected within each root system. Already in the first stage of succession, a high fungal diversity was present in the B. vivipara root systems. Total number of OTUs increased significantly along the gradient towards climax vegetation, but the average number of OTUs per root system stayed unchanged. There was a high patchiness in distribution of fungal OTUs across root systems, indicating that stochastic processes to a large extent structure the fungal communities. However, time since deglaciation had impact on the fungal community structure, as a systematic shift in the community composition was observed along the chronosequence. Ectomycorrhizal basidiomycetes were the dominant fungi in the roots of B. vivipara, when it comes to both number of OTUs and number of sequences.