High-fat diet-induced obesity leads to increased NO sensitivity of rat coronary arterioles: role of soluble guanylate cyclase activation

The impact of obesity on nitric oxide (NO)-mediated coronary microvascular responses is poorly understood. Thus NO-mediated vasomotor responses were investigated in pressurized coronary arterioles ( approximately 100 microm) isolated from lean (on normal diet) and obese (fed with 60% of saturated fat) rats. We found that dilations to acetylcholine (ACh) were not significantly different in obese and lean rats (lean, 83 +/- 4%; and obese, 85 +/- 3% at 1 microM), yet the inhibition of NO synthesis with N(omega)-nitro-l-arginine methyl ester reduced ACh-induced dilations only in vessels of lean controls. The presence of the soluble guanylate cyclase (sGC) inhibitor oxadiazolo-quinoxaline (ODQ) elicited a similar reduction in ACh-induced dilations in the two groups of vessels (lean, 60 +/- 11%; and obese, 57 +/- 3%). Dilations to NO donors, sodium nitroprusside (SNP), and diethylenetriamine (DETA)-NONOate were enhanced in coronary arterioles of obese compared with lean control rats (lean, 63 +/- 6% and 51 +/- 5%; and obese, 78 +/- 5% and 70 +/- 5%, respectively, at 1 microM), whereas dilations to 8-bromo-cGMP were not different in the two groups. In the presence of ODQ, both SNP and DETA-NONOate-induced dilations were reduced to a similar level in lean and obese rats. Moreover, SNP-stimulated cGMP immunoreactivity in coronary arterioles and also cGMP levels in carotid arteries were enhanced in obese rats, whereas the protein expression of endothelial NOS and the sGC beta1-subunit were not different in the two groups. Collectively, these findings suggest that in coronary arterioles of obese rats, the increased activity of sGC leads to an enhanced sensitivity to NO, which may contribute to the maintenance of NO-mediated dilations and coronary perfusion in obesity.     

Multistep mass tagging coupled with 2D LC-MS--an approach to increasing the number of identified proteins.

Identification of large numbers of proteins from complex biological samples is a continuing challenge in the area of quantitative proteomics. We introduce here a simple and reliable multistep mass tagging technique using our recently developed solid phase mass tagging reagents. When coupled with two-dimensional liquid chromatography/nano-electrospray ionization ion trap mass spectrometry (2D-LC/nano-ESI-MS), this method allows enhanced protein identification when tested on samples from prokaryotic and eukaryotic sources. The proteome of Escherichia coli D21 grown to either mid-exponential or stationary phase, and the membrane proteome from established breast cancer cell lines BT474 and MCF7 were used as model systems in these experiments. In both experiments, the numbers of total identified proteins are at least twice the numbers identified from a single tagging cycle. The sample complexity can be effectively reduced with corresponding increases in protein identification using the multistep method. The strategy described here represents a potentially powerful technique for large-scale qualitative and quantitative proteome research.     

Endorphin excess at weaning durably influences sexual activity, uterine estrogen receptor's binding capacity and brain serotonin level of female rats.

Perinatally, the first encounter between the maturing receptor and its target hormone results in hormonal imprinting, which adjusts the binding capacity of the receptor for life. In the presence of an excess of the target hormone or foreign molecules than can be bound by the receptor, faulty imprinting carries life-long consequences. In cytogenic organs, imprinting could also be provoked in other periods of life (late imprinting). Imprinting also durably influences the production of the imprinter and related hormones. In the present study, single beta-endorphin doses was given to three-week old female rats at 3 microg/animal, and the serotonin in five brain regions (frontal cortex, striatum, hippocampus, hypothalamus and brain stem) and uterine estrogen receptor content were determined, thymic glucocorticoid receptor binding capacity was measured, and sexual behavior was tested at five months of age. Brain serotonin levels highly significantly decreased, while sexual activity (Meyerson index and lordosis quotient) increased. At the same time, uterine estrogen receptor affinity decreased. There was no change in receptor binding capacity in the thymus. We will go on to discuss interrelations between the results. The experiments demonstrate that a non-perinatal treatment with a molecule acting at receptor level (late imprinting) can also lastingly influence various indexes in non-cytogenic organs. The results call attention to the possible long-lasting influence of an endorphin surge (caused, for example, by pain) on brain serotonin content and sexual behavior.     

A wireless communication system for determining site fidelity of juvenile dusky groupers Epinephelus marginatus (Lowe, 1834) using coded acoustic transmitters

The value of marine reserves to protect threatened and vulnerable species, such as the dusky grouper (Epinephelus marginatus), is often uncertain because of a lack of knowledge about their ecology and habitat utilisation. To address this issue, a Wireless Communication System for location-based positioning of fish tagged with coded acoustic transmitters was installed to determine site fidelity of juvenile dusky groupers (mean total length = 30.4 cm, range: 20.5–40.0 cm; mean total weight = 470 g, range: 113–950 g) within a marine reserve (Island of Ustica, Sicily, Italy). The system was based on five wireless hydrophones distributed approximately 200 m apart along the coastline of the reserve, each monitoring acoustic signals from transmitters within a reception cell. The hydrophones retransmitted detected signals as a radio signal to a land-based receiver and data logger. Unique identifiers for each hydrophone permitted the identification of the detection cell containing the tagged fish. The wireless hydrophones had partly overlapping detection ranges, dividing the marine reserve into more than five different detection cells. The average detection range was 232 m, but coverage varied from site to site and with time, depending on many factors, such as ambient noise, wave action and signal strength of the transmitters. Despite some limitations, the Wireless Hydrophone System (WHS) combined with manual tracking provided a valuable tool for investigating site fidelity in the dusky grouper. Preliminary home ranges for the fish were estimated by the kernel method (probability level of 95%), based on a limited set of manual tracking data, to be a median 5312 m² (range 1673–18626 m²).     

European Cuckoo Cuculus canorus parasitism and host's rejection behaviour in a heavily parasitized Great Reed Warbler Acrocephalus arundinaceus population

An unusually high frequency (64%) of European Cuckoo Cuculus canorus parasitism was found in Great Reed Warbler Acrocephalus arundinaceus clutches in central Hungary. Sixty-four per cent of the parasitized clutches contained one Cuckoo egg, 23% contained two, 10% had three and 3% had four. This means that 58% of the Cuckoo eggs were found in multiply parasitized clutches. In multiple parasitism the laying second Cuckoo removed an egg from the clutch randomly, so preferred neither the host eggs, nor the concurrent Cuckoo egg. Host response towards the parasitic eggs showed 66% acceptance, 12% ejection, 20% desertion and 2% egg burial. We found great variation in both the host and the parasitic egg colour and pattern. This reduces the chance that the parasitic egg's appearance matched that of the hosts' but, in spite of this, almost perfect mimesis was found in 28% of the Cuckoo eggs. Poorly mimetic Cuckoo eggs were more frequently rejected by Great Reed Warblers than parasite eggs that were very similar to the host eggs. This high level of mimicry sometimes makes it difficult for the observer to identify the parasitic egg, especially when it is similar in size to the host eggs. It is also difficult for the host, as shown by the relatively high recognition error and ejection cost.     

Littoral macrophyte–periphyton complexes in two Hungarian shallow waters

Periphyton developing on the surfaces of emergent and submerged aquatic plants has a significant influence on water quality. The periphyton types that form on various plant species can be characterized by their mass values, the proportion of the present organic and inorganic fractions, as well as their chlorophyll-a contents. Studies on periphyton complexes constituting integrated biomonitoring systems are useful to gain essential long-term information about the performance of shallow water bodies. The filtering and settling effect of Phragmites and other aquatic plants, as well as their periphyton was examined and clearly observable in the water areas and non-flooded aquatic habitats belonging to the second phase of Kis-Balaton Protection System, as it was indicated by the mass values and ash contents. The periphyton forming on the aquatic vegetation that annually develops in Kisköre Reservoir and yields a considerable biomass has a critical part in influencing water quality. The only difference (p<0.05) was found in the ash content of the periphyton, being lower in Kis-Balaton (48.64 ± 2.29 S.E., %) and higher in Kisköre Reservoir (57.42 ± 2.54 S.E., %). This paper presents the dry mass of the periphyton, as well as its ash and chlorophyll-a content, and the results obtained on the composition of the alga species of the periphyton.

     

Direct chemotactic effect of bradykinin and related peptides-significance of amino- and carboxyterminal character of oligopeptides in chemotaxis of tetrahymena pyriformis.

The chemotactic character of the nonapeptide bradykinin (BK1-9) and its derivatives was studied in the eukaryotic ciliated model Tetrahymena pyriformis. The results demonstrate that BK1-9 has a direct and ligand-specific chemoattractant effect (maximal at 10(-11) m) without any intermediate substance as is essential in some mammalian test systems. Evaluation of the chemotactic effect elicited by derivatives showed that the presence of N- and C-terminal arginines can influence chemotactic potency of the molecule via expression of pyrrolidine and aromatic ring structures of terminal amino acid residues. Removal of the N-terminal Arg (expression of Pro) results in a significant decrease in chemotaxis (BK2-9), while further truncation of the C-terminal, causing expression of the aromatic ring of Phe (BK2-8), results in a highly chemoattractant variant. A single pyrrolidine ring on the C-terminus BK1-7 also has a positive effect on the chemotactic character, however further truncation (BK1-6, BK1-5) causes the chemoattractant character to become chemorepellent. Study of chemotactic selection with BK derivatives supports our previous findings that only phylogenetically selected ligands or their close derivatives are able to induce long-term selection with chemotaxis.     

Mitochondrial Dehydrogenases in Different Taxa of Tetrahymena: Effect of Insulin

Mitochondrial dehydrogenase activity was measured in seven taxa of Tetrahymena (T. pyriformis G1, T. hegewishi, T. malaccensis, T. pigmentosa, T. shapiro, T. thermophila CU-399, T. thermophila MS-1). Enzyme activity was different in the taxa investigated. Insulin reduced enzyme activity in six of the seven taxa studied. The duration of activity reduction was relatively long (5–10 min.) in most of the cases, and in T. hegewishi this lasted up to the end of the measurements (30 min.). There was no interrelation between the basic dehydrogenase activity of the taxon and the effect of insulin. There was also no correlation between the degree of relationship (of the taxa) and the dehydrogenase profile after insulin treatment.     

Receptor amplifying effect of serotonin and serotonin analogues in a protozoan (Tetrahymena) model system

A 72-hour treatment of Tetrahymena with serotonin analogues at a concentration of 10(-9) mmole/l resulted in a decrease of the reproduction rate, whereas serotonin itself was ineffective. On the second exposure to antagonists, cellular division was enhanced but its rate remained around or below the control value, while the second exposure to serotonin produced a more marked effect than the first one did, which finding implies a receptor amplification. The most pronounced receptor amplification was caused by the first exposure to serotonin. This observation indicates that the receptor is rather selective as regards the amplification effect.