In vitro dose-response effects of poly(amidoamine) dendrimers [amino-terminated and surface-modified with N-(2-hydroxydodecyl) groups] and quantitative determination by a liquid chromatography-hybrid quadrupole/time-of-flight mass spectrometry based method.

This article presents a dose-response study of the effects of two types of third-generation (G3) and fourth-generation poly(amidoamine) (PAMAM) dendrimers on two cell lines (RTG-2 and H4IIE) by in vitro cytotoxicity assays with 3-(4,5-dimethylthizol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), neutral red uptake (NRU), and lactate dehydrogenase (LDH) assays. We particularly investigated the potential cytotoxic effect of positive surface charge, which a cationic amino-terminated PAMAM dendrimer can display, on the marked ability of PAMAM dendrimers to cross the cell membrane compared with PAMAM dendrimers functionalized with chains of N-(2-hydroxydodecyl). Quantification of dose-response effects was performed by use of mass spectrometry analysis. The analytical method using liquid chromatography-hybrid quadrupole/time-of-flight mass spectrometry that we developed allowed characterization of defective dendrimers instead of "ideal structures." Identification was based on accurate mass measurement, assignment of elemental composition, and the fully resolved (13)C/(12)C isotopic clusters of the multiply charged ions of PAMAM dendrimers. Validation of the liquid chromatography-mass spectrometry method made possible reliable and reproducible quantification of the extracellular and intracellular concentration of dendrimers at a micromolar level (limits of detection from 0.14 to 1.34 µM and from 0.43 to 1.82 µM in standard and culture medium, respectively). A higher cytotoxicity was found with the H4IIE cell line for surface-modified PAMAM dendrimers. The LDH assay was significantly more sensitive than the MTT and NRU assays, with half-maximal inhibitory concentrations (IC(50)) of 12.96 and 38.31 µg mL(-1) for surface-modified G3 and G4 dendrimers, respectively. No cytotoxic effects, in terms of IC(50), of amino-terminated PAMAM dendrimers were observed on both H4IIE and RTG-2 cells when the concentration was below 500 µg mL(-1) for G3 and G4 dendrimers.

Effects of cerium oxide nanoparticles to fish and mammalian cell lines: An assessment of cytotoxicity and methodology.

Two cerium oxide nanoparticles (CeO(2) NPs) and one micro-sized CeO(2) particle were thoroughly characterized in their pristine form, in water and in cell culture medium. The particles were tested for cytotoxicity to the H4IIE rat hepatoma cell line or the RTG-2 rainbow trout gonadal cell line by means of four standard cytotoxicity assays. Nominal concentrations were verified by inductively coupled plasma mass spectrometry (ICP-MS) and methods were assessed for their suitability to detect reliably adverse effects due to particle exposure. All three particles showed aggregation in water and media. In the H4IIE cell line, the MTT cytotoxicity test revealed that negative effects could be observed for the CeO(2) NPs after 24h and for all particles after 72h of exposure, making the effects size, concentration and time dependent. No negative effect for the concentrations tested was detected for the remaining three assays and the RTG-2 cell line, making the MTT assay and the H4IIE cell line an appropriate system to assess adverse effects of CeO(2) NPs. A verification of the nominal concentration through ICP-MS revealed that there was a discrepancy between nominal and measured concentration depending on concentration and particle tested. Interferences of particles with assays were found to be present and need to be taken into consideration.

Varroa-tolerant Italian honey bees introduced from Brazil were not more efficient in defending themselves against the mite Varroa destructor than Carniolan bees in Germany

In Europe and North America honey bees cannot be kept without chemical treatments against Varroa destructor. Nevertheless, in Brazil an isolated population of Italian honey bees has been kept on an island since 1984 without treatment against this mite. The infestation rates in these colonies have decreased over the years. We looked for possible varroa-tolerance factors in six Italian honey bee colonies prepared with queens from this Brazilian island population, compared to six Carniolan colonies, both tested at the same site in Germany. One such factor was the percentage of damaged mites in the colony debris, which has been reported as an indicator of colony tolerance to varroa. A mean of 35.8 of the varroa mites collected from the bottoms of the Italian bee colonies were found damaged, among which 19.1 were still alive. A significantly greater proportion of damaged mites were found in the Carniolan bees (42.3) and 22.5 were collected alive. The most frequent kind of damage found was damaged legs alone, affecting 47.4 of the mites collected from debris in Italian bees, which was similar to the amount found in Carniolan colonies (46). The mean infestation rate by the varroa mite in the worker brood cells in the Italian bee colonies was 3.9 in June and 3.5 in July, and in drone brood cells it was 19.3 in June. In the Carniolan honey bee colonies the mean infestation rates in worker brood cells were 3.0 and 6.7, respectively in the months of June and July and 19.7 in drone brood cells in June. In conclusion, the 'Varroa-tolerant' Italian honey bees introduced from Brazil produced lower percentages of damaged mites (Varroa destructor) in hive debris and had similar brood infestation rates when compared to 'susceptible' Carniolan bees in Germany. In spite of the apparent adaptation of this population of Italian bees in Brazil, we found no indication of superiority of these bees when we examined the proportions of damaged mites and the varroa-infestation rates, compared to Carniloan bees kept in the same apiary in Germany

Varroa-tolerant Italian honey bees introduced from Brazil were not more efficient in defending themselves against the mite Varroa destructor than Carniolan bees in Germany.

In Europe and North America honey bees cannot be kept without chemical treatments against Varroa destructor. Nevertheless, in Brazil an isolated population of Italian honey bees has been kept on an island since 1984 without treatment against this mite. The infestation rates in these colonies have decreased over the years. We looked for possible varroa-tolerance factors in six Italian honey bee colonies prepared with queens from this Brazilian island population, compared to six Carniolan colonies, both tested at the same site in Germany. One such factor was the percentage of damaged mites in the colony debris, which has been reported as an indicator of colony tolerance to varroa. A mean of 35.8% of the varroa mites collected from the bottoms of the Italian bee colonies were found damaged, among which 19.1% were still alive. A significantly greater proportion of damaged mites were found in the Carniolan bees (42.3%) and 22.5% were collected alive. The most frequent kind of damage found was damaged legs alone, affecting 47.4% of the mites collected from debris in Italian bees, which was similar to the amount found in Carniolan colonies (46%). The mean infestation rate by the varroa mite in the worker brood cells in the Italian bee colonies was 3.9% in June and 3.5% in July, and in drone brood cells it was 19.3% in June. In the Carniolan honey bee colonies the mean infestation rates in worker brood cells were 3.0 and 6.7%, respectively in the months of June and July and 19.7% in drone brood cells in June. In conclusion, the 'Varroa-tolerant' Italian honey bees introduced from Brazil produced lower percentages of damaged mites (Varroa destructor) in hive debris and had similar brood infestation rates when compared to 'susceptible' Carniolan bees in Germany. In spite of the apparent adaptation of this population of Italian bees in Brazil, we found no indication of superiority of these bees when we examined the proportions of damaged mites and the varroa-infestation rates, compared to Carniloan bees kept in the same apiary in Germany.

Mandibular Gland Volatiles and Their Ontogenetic Patterns in Queen Honey Bees, Apis mellifera carnica.

Extracts of mandibular glands taken from adult queens of the honey bee, Apis mellifera carnica, were analysed by gas chromatography-mass spectroscopy. More than 100 compounds could be identified among which oxygenated fatty acids with six, eight, 10 and 12 carbon atoms are particularly interesting since they show structural relationships to the queen substance, (E)-9-oxo-2-decenoic acid. Changes in the patterns of volatiles were followed up from emergence until the full dominant status of an egg-laying queen in a strong colony. Generally, the amount of volatiles per gland was found to increase with age. The final level of queen substance (9-ODA) content is reached at the postmating stage about 10 days after emergence. Ontogenetic patterns of concentrations were determined for those components regarded to predominantly contribute to the royal pheromone. Characteristic compositions of signals, possibly involved in the premating, mating and postmating dominance status of a honey bee queen are discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved

Juvenile hormone titer in capped worker brood of Apis mellifera and reproduction in the bee mite Varroa jacobsoni.

Juvenile hormone (JH) titers were recorded from fifth instar worker larvae of Apis mellifera carnica, Apis mellifera lamarckii, and Africanized honeybees kept under temperate and tropical climatic conditions. No differences in hormone titer according to honeybee race or climatic conditions were determined. However, the rate of reproduction of the ectoparasitic mite, Varroa jacobsoni, on larvae of the different honeybee races was highly variable. The possible role of honeybee JH in control of the parasite's reproduction is discussed.

Septic arthritis caused by Bacteroides fragilis.

As improvements in bacteriologic techniques have enhanced the recovery of anaerobic bacteria from clinical specimens, there has been an increasing awareness of the role of anaerobes in disease. Bacteroides fragilis is the most common anaerobic organism found in clinical specimens. Although it is the anaerobe most frequently associated with bacteremia and a common isolate in intraabdominal infections, infections of the female genital tract, wounds, and abscesses, B. fragilis is a rare cause of septic arthritis. The isolation of this organism from four patients with septic arthritis in three Cleveland hospitals between 1978 and 1982 suggests that septic arthritis due to B. fragilis may be a more common clinical entity than previously appreciated. In this report we describe these cases and review the pertinent literature.

Intravenous and standard immune serum globulin preparations interfere with uptake of 125I-C3 onto sensitized erythrocytes and inhibit hemolytic complement activity.

Antibody-sensitized sheep erythrocytes were used as a model to determine the effects of therapeutic immune serum globulin (ISG) preparations on the ability of this particulate activator to fix C3 and initiate hemolysis. Both standard and intravenous forms of ISG inhibit uptake of 125I-C3, presumably by competing for the deposition of "nascent" C3b molecules onto the erythrocytes. Both forms of ISG also inhibit hemolytic activity of whole serum or purified complement components. The inhibition appears to be a specific property of IgG itself, since similar inhibition was not caused by equivalent concentrations of human serum albumin, and was not affected by the buffer in which the ISG was dissolved. Interference with C3 uptake onto antibody-sensitized platelets and/or inhibition of hemolytic complement activity could contribute to the efficacy of high dose intravenous ISG in idiopathic thrombocytopenic purpura.

Inhibition of DNA synthesis accompanied by stimulation of protein synthesis in novobiocin-treated developing sea urchins.

Novobiocin, an inhibitor of bacterial DNA gyrase, induces abnormalities in developing embryo of the American sea urchin Arbacia punctulata. In addition this drug is also a specific inhibitor of DNA synthesis. This block in DNA synthesis is accompanied by a stimulation in protein synthesis. This effect appears to be characteristic of novobiocin, as hydroxyurea another inhibitor of DNA synthesis in the developing sea urchin, does not stimulate protein production. The present findings constitute the first report of a novobiocin-induced specific inhibition of DNA synthesis in a higher eukaryote.

Microwave spectroscopic imagery of the Earth.

The microwave spectrometer on the Nimbus 6 satellite has produced the first microwave spectral images of the earth. It has yielded global maps of (i) atmospheric temperature profiles, (ii) the distributions of water vapor and liquid water over ocean, and (iii) the coverage and type of ice and snow. The method has potential for operational synoptic monitoring.

On the mechanism of the acridine orange sensitized photodynamic inactivation of lysozyme. II. Kinetics in presence of N-acetylglucosamine.

The photodynamic deactivation of lysozyme in presence of acridine orange is caused by a reaction between singlet oxygen formed via the dye triplet state and the protein. In order to identify the region where the singlet oxygen reacts with the protein we have investigated the kinetics of the deactivation in presence ofthe inhibitor of the enzymatic reaction N-acetylglucosamine (GlcNAc). The overall experimental rate constant becomes slower with increasing saccharide concentrations. As we can exclude experimentally that this kinetical effect is caused in presence of the saccharide by a physical quenching of singlet oxygen or of the dye triplet state it has to be assumed that GlcNAc protects the surrounding of its bindings place at subsite C of the enzymatic center sterically against an attack of singlet oxygen. In this region three tryptophan residues are located, which could be sensitive against singlet oxygen. Surprisingly, however, it has been found that only those species are protected, in which a second saccharide molecule is bound to the protein, probably at subsite E at the enzymatic center, where no sensitive amino acid side chains are located.

On the mechanism of the acridine orange sensitized photodynamic inactivation of lysozyme. I. Basic kinetics.

The kinetics of the photodynamic desactivation of lysozyme in presence of acridine orange as the sensitizer have been investigated in detail varying oxygen, protein, dye concentration, ionic strength and pH value. The kinetics can be approximately described as an over all pseudo-first-order rate process. Changing the solvent from water to D2O or by quenching experiments in presence of azide ions it could be shown that the desactivation of lysozyme is caused exclusively by singlet oxygen. The excited oxygen occurs via the triplet state of the dye with a rate constant considerably lower than that to be expected for a diffusionally controlled reaction. Singlet oxygen react chemically (desactivation, k=2.9 X 10(7) m(-1) sec(-1)) and physically (quenching process, k=4.1 X 10(8) m(-1) sec(-1)) with the enzyme. The kinetical analysis shows that additional chemical reaction between singlet oxygen and lysozyme would have only little influence on the kinetics of the desactivation as long as their products would be enzymatically active and their kinetical constants would be less than about 1 X 10(8) m(-1) sec(-1)).

Microwave spectrometer on the nimbus 5 satellite: meteorological and geophysical data.

The Nimbus 5 microwave spectrometer has been used to measure thermal radiation in five frequency bands between 22.235 and 58.8 gigahertz, and has yielded both the temperature profile and, over ocean, the vapor and liquid water content of the terrestrial atmosphere, even in overcast conditions. Information has also been obtained on geophysical parameters that affect the surface emissivity, such as ice type, sea roughness, and snow cover. The experiment demonstrates the considerable potential of passive microwave sensing of meteorological and geophysical parameters.