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Intensive algal cultivation usually requires a high flux of dissolved inorganic carbon (Ci) to support productivity, particularly for high density algal cultures. Carbon dioxide (CO2) enrichment can be used to overcome Ci limitation and enhance productivity of algae in intensive culture, however, it is unclear whether algal species with the ability to utilise bicarbonate (HCO3 (-)) as a carbon source for photosynthesis will benefit from CO2 enrichment. This study quantified the HCO3 (-) affinity of three green tide algal species, Cladophora coelothrix, Cladophora patentiramea and Chaetomorpha linum, targeted for biomass and bioenergy production. Subsequently, we quantified productivity and carbon, nitrogen and ash content in response to CO2 enrichment. All three species had similar high pH compensation points (9.7-9.9), and grew at similar rates up to pH 9, demonstrating HCO3 (-) utilization. Algal cultures enriched with CO2 as a carbon source had 30% more total Ci available, supplying twenty five times more CO2 than the control. This higher Ci significantly enhanced the productivity of Cladophora coelothrix (26%), Chaetomorpha linum (24%) and to a lesser extent for Cladophora patentiramea (11%), compared to controls. We demonstrated that supplying carbon as CO2 can enhance the productivity of targeted green tide algal species under intensive culture, despite their clear ability to utilise HCO3 (-).

The optimised reduction of dissolved nutrient loads in aquaculture effluents through bioremediation requires selection of appropriate algal species and strains. The objective of the current study was to identify target species and strains from the macroalgal genus Ulva for bioremediation of land-based aquaculture facilities in Eastern Australia. We surveyed land-based aquaculture facilities and natural coastal environments across three geographic locations in Eastern Australia to determine which species of Ulva occur naturally in this region and conducted growth trials at three temperature treatments on a subset of samples from each location to determine whether local strains had superior performance under local environmental conditions. DNA barcoding using the markers ITS and tufA identified six species of Ulva, with U. ohnoi being the most common blade species and U. sp. 3 the most common filamentous species. Both species occurred at multiple land-based aquaculture facilities in Townsville and Brisbane and multiple strains of each species grew well in culture. Specific growth rates of U. ohnoi and U. sp. 3 were high (over 9% and 15% day(-1) respectively) across temperature treatments. Within species, strains of U. ohnoi had higher growth in temperatures corresponding to local conditions, suggesting that strains may be locally adapted. However, across all temperature treatments Townsville strains had the highest growth rates (11.2-20.4% day(-1)) and Sydney strains had the lowest growth rates (2.5-8.3% day(-1)). We also found significant differences in growth between strains of U. ohnoi collected from the same geographic location, highlighting the potential to isolate and cultivate fast growing strains. In contrast, there was no clearly identifiable competitive strain of filamentous Ulva, with multiple species and strains having variable performance. The fast growth rates and broad geographical distribution of U. ohnoi make this an ideal species to target for bioremediation activities at land-based aquaculture facilities in Eastern Australia.

This paper is a review of local plants used in water infusions as aromatic and refreshing hot beverages (recreational tea) consumed in food-related settings in Europe, and not for specific medicinal purposes. The reviewed 29 areas are located across Europe, covering the post-Soviet countries, eastern and Mediterranean Europe. Altogether, 142 taxa belonging to 99 genera and 40 families were reported. The most important families for making herbal tea in all research areas were Lamiaceae and Asteraceae, while Rosaceae was popular only in eastern and central Europe. With regards to botanical genera, the dominant taxa included Mentha, Tilia, Thymus, Origanum, Rubus and Matricaria. The clear favorite was Origanum vulgare L., mentioned in 61% of the regions. Regionally, other important taxa included Rubus idaeus L. in eastern Europe, Chamaemelum nobile (L.) All. in southern Europe and Rosa canina L. in central Europe. Future research on the pharmacological, nutritional and chemical properties of the plants most frequently used in the tea-making process is essential to ensure their safety and appropriateness for daily consumption. Moreover, regional studies dedicated to the study of local plants used for making recreational tea are important to improve our understanding of their selection criteria, cultural importance and perceived properties in Europe and abroad.

Background: The idea that knowledge flows through social networks is implicit in research on traditional knowledge, but researchers have paid scant attention to the role of social networks in shaping its distribution. We bridge those two bodies of research and investigate a) the structure of network of exchange of plant propagation material (germplasm) and b) the relation between a person's centrality in such network and his/her agroecological knowledge., Methods: We study 10 networks of germplasm exchange (n = 363) in mountain regions of the Iberian Peninsula. Data were collected through participant observation, semi-structured interviews, and a survey., Results: The networks display some structural characteristics (i.e., decentralization, presence of external actors) that could enhance the flow of knowledge and germplasm but also some characteristics that do not favor such flow (i.e., low density and fragmentation). We also find that a measure that captures the number of contacts of an individual in the germplasm exchange network is associated with the person's agroecological knowledge., Conclusion: Our findings highlight the importance of social relations in the construction of traditional knowledge.

Although 1,2-cyclic sulfamidates derived from α-methylisoserine undergo nucleophilic displacement at the quaternary center, to the best of our knowledge their behavior with amines as nucleophiles has never been explored. We have found that a broad range of amines can be used, demonstrating the scope of the reaction, and that excellent control of the chemoselectivity can be achieved. Application of this methodology for the synthesis of a chiral α,β-diamino acid and an important piperazinone heterocycle is also presented. Additionally, we have found that DMF and DMSO behave not only as polar aprotic solvents but also as O-nucleophilic reagents, allowing the incorporation of an oxygen atom at a quaternary center of the electrophile, with inversion of configuration., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The interactions between the fruit fly Anastrepha spatulata Stone (Diptera: Tephritidae) and two species of moths, the gelechiid Coleotechnites sp. and an unidentified tortricid species, were examined on their effects on seed production in terms of their exploitation within fruits of Schoepfia schreberi J.F. Gmel (Santalales: Olacaceae). The study was carried out in three experimental sites during 3 yr. Under conditions of abundant fruit, A. spatulata was the dominant exploiter at the population level, as shown by its ability to infest the largest number of fruits of the three herbivores and substantially displace its moth competitors. In a separate experiment, when resource-partitioning species were excluded, A. spatulata infested twice as many fruits as the two moth species (44.3%). Field observations examined the ability of A. spatulata to locate suitable fruits. We found that, the variation in fruit characters influenced fruit suitability in relation to its size (4.1261 ± 0.0272 mm [mean ± SE]) and weight (0.0618 ± 0.0005 mg [mean ± SE]). Uninfested fruits produce viable seed but the interaction of all species (exploiting for the fruits) led to limited seed formation.

An interlaboratory collaborative study to validate a colorimetric phosphatase inhibition assay for quantitative determination of the okadaic acid (OA) toxins group in molluscs, OkaTest, was conducted. Eight test materials, including mussels, scallops, clams, and cockles, were analyzed as blind duplicates. Blank samples and materials containing different OA toxin levels ranging from 98 to 275 microg/kg OA equivalents were included. The study was carried out by a total of 16 laboratories from 11 different countries. Values obtained for repeatability relative standard deviations (RSDr) ranged from 5.4 to 11.2% (mean 7.5%). Reproducibility RSD (RSD(R)) values were between 7.6 and 13.2% (mean 9.9%). The Horwitz ratio (HorRat) values ranged between 0.4 and 0.6. A recovery assay was also carried out using a sample spiked with OA. A mean recovery of 98.0% and an RSD of 14.5% were obtained. The results obtained in this validation study indicate that the colorimetric phosphatase inhibition assay, OkaTest, is suitable for quantitative determination of the OA toxins group. OkaTest could be used as a test that is complementary to the reference method for monitoring the OA toxins group.

The reactions of cyclic sulfamidates as electrophiles with a variety of nitrogen-containing aromatic heterocycle nucleophiles, such as pyridines, N-alkylimidazoles and N-methylbenzimidazol, was explored. In all cases, although the nucleophilic substitution reactions occurred on quaternary centres, elimination products were not detected. The inversion of configuration at this quaternary centre was determined by X-ray diffraction analysis and the enantiomeric excess of the reactions was checked by chiral HPLC. This synthetic approach allowed us to obtain a new family of chiral charged β(2,2)-amino acids, including a new bisamino acid that incorporates an imidazolium salt as a cross-linker. In this context, the treatment of these chiral imidazolium salts with Ag(2)O opens the way to new chiral N-heterocyclic carbenes, which are important substrates in the fields of organometallic and organocatalytic chemistry. Additionally, we have done a thorough conformational analysis of these β-amino acid derivatives, both in the solid state and in solution. The most important conformational feature of these acyclic systems is the rigidity of the N-CH(2)-C-N(+) dihedral angle, which is essentially due to the gauche effect., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Aqueous extracts from common tropical seaweeds were evaluated for their effect on the life cycle of the commercially important ectoparasite, Neobenedenia sp. (Platyhelminthes: Monogenea), through the survival of attached adult parasites, period of embryonic development, hatching success and oncomiracidia (larvae) infection success. There was no significant effect of any extract on the survival of adult parasites attached to fish hosts or infection success by oncomiracidia. However, the extracts of two seaweeds, Ulva sp. and Asparagopsis taxiformis, delayed embryonic development and inhibited egg hatching. The extract of A. taxiformis was most effective, inhibiting embryonic development of Neobenedenia sp. and reducing hatching success to 3% compared with 99% for the seawater control. Furthermore, of the 3% of eggs that hatched, time to first and last hatch was delayed (days 14 and 18) compared with the seawater control (days 5 and 7). Asparagopsis taxiformis shows the most potential for development as a natural treatment to manage monogenean infections in intensive aquaculture with the greatest impact at the embryo stage., (Copyright © 2012 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

Some morphogenetic and metabolic processes were sensitive to a high atmospheric CO(2) concentration during sunflower primary leaf ontogeny. Young leaves of sunflower plants growing under elevated CO(2) concentration exhibited increased growth, as reflected by the high specific leaf mass referred to as dry weight in young leaves (16 days). The content of photosynthetic pigments decreased with leaf development, especially in plants grown under elevated CO(2) concentrations, suggesting that high CO(2) accelerates chlorophyll degradation, and also possibly leaf senescence. Elevated CO(2) concentration increased the oxidative stress in sunflower plants by increasing H(2)O(2) levels and decreasing activity of antioxidant enzymes such as catalase and ascorbate peroxidase. The loss of plant defenses probably increases the concentration of reactive oxygen species in the chloroplast, decreasing the photosynthetic pigment content as a result. Elevated CO(2) concentration was found to boost photosynthetic CO(2) fixation, especially in young leaves. High CO(2) also increased the starch and soluble sugar contents (glucose and fructose) and the C/N ratio during sunflower primary leaf development. At the beginning of senescence, we observed a strong increase in the hexoses to sucrose ratio that was especially marked at high CO(2) concentration. These results indicate that elevated CO(2) concentration could promote leaf senescence in sunflower plants by affecting the soluble sugar levels, the C/N ratio and the oxidative status during leaf ontogeny. It is likely that systemic signals produced in plants grown with elevated CO(2), lead to early senescence and a higher oxidation state of the cells of these plant leaves., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

Context: Pancreatic injuries caused by blunt trauma are often treated conservatively, except for the highest grades of these., Case Report: We report a case of complete transection of the distal pancreas in a young adult which was successfully managed by spleen-preserving laparoscopic distal pancreatectomy followed by an islet autotransplantation in the patient's forearm striated muscle., Conclusion: We describe a mini-invasive approach for pancreatectomy with restoration of resected islets to the patient.

A phosphatase inhibition assay for detection of okadaic acid (OA) toxins in shellfish, OkaTest, was single laboratory validated according to international recognized guidelines (AOAC, EURACHEM). Special emphasis was placed on the ruggedness of the method and stability of the components. All reagents were stable for more than 6 months and the method was highly robust under normal laboratory conditions. The limit of detection and quantification were 44 and 56 µg/kg, respectively; both below the European legal limit of 160 µg/kg. The repeatability was evaluated with 2 naturally contaminated samples. The relative standard deviation (RSD) calculated was 1.4% at a level of 276 µg/kg and 3.9% at 124 µg/kg. Intermediate precision was estimated by testing 10 different samples (mussel and scallop) on three different days and ranged between 2.4 and 9.5%. The IC(50) values of the phosphatase used in this assay were determined for OA (1.2 nM), DTX-1 (1.6 nM) and DTX-2 (1.2 nM). The accuracy of the method was estimated by recovery testing for OA (mussel, 78-101%; king scallop, 98-114%), DTX-1 (king scallop, 79-102%) and DTX-2 (king scallop, 93%). Finally, the method was qualitatively compared to the mouse bioassay and LC-MS/MS.

We tested how the availability of carbon and nitrogen determines both the production of Asparagopsis taxiformis (Delile) V. Trevis. and content of the two major halocarbons, bromoform and dibromoacetic acid. The halogenated secondary metabolites of Asparagopsis species are particularly interesting from an applied perspective due to their remarkable antimicrobial activity. Terrestrial ecologists named the relationship between resources and secondary metabolites as the carbon (C)/nutrient balance (CNB) hypothesis. This relationship was tested both in the laboratory, with a factorial analysis using different concentrations of total ammonia (TAN) and dissolved inorganic carbon (DIC), and in an integrated aquaculture system where TAN and DIC fluxes of fish effluent were manipulated. The total C/N content of A. taxiformis biomass cultivated in laboratory was highly significantly linearly related to the content of both halocarbons, as predicted by the CNB hypothesis. A. taxiformis cultivated at low levels of carbon and high levels of nitrogen (N) (lowest C/N ratio) had the lowest content in both halogenated metabolites. Increased availability of CO2 in the medium resulted in a general higher halocarbon content in the biomass, even though the effect was only statistically significant for bromoform at high levels of N. The farm experiments supported the results of the laboratory experiments. DIC fluxes had the highest effect on the production of both bromoform and biomass, as shown by multiple regression analysis. In A. taxiformis integrated aquaculture, C, rather than N, is the most important factor affecting the production of biomass and of valuable halocarbon secondary metabolites., (© 2011 Phycological Society of America.)

This paper explores the role of a hindered cyclic sulfamidate derived from α-methylisoserine as an electrophile in a nucleophilic displacement reaction with nitrogen-containing aromatic heterocycles. Several imidazoles and pyrazole were tested as nucleophiles in the absence of an additional base to give the corresponding ring-opening compounds. We show that the process takes place by inversion of the configuration of the quaternary electrophilic center, retaining the enantiomeric excess of the starting sulfamidate. This reaction opens the way to obtain important quaternary imidazole derivatives such as an innovative type of bis-amino acid related to histidinoalanine and a novel α,α-disubstituted β-amino acid (β(2,2)-amino acid).

Background: There is a lack of prospective studies evaluating the natural history of colonic ischaemia (CI). We performed such a study to evaluate the clinical presentation, outcome, and mortality as well as clinical variables associated with poor prognosis., Methods: An open, prospective, and multicentre study was conducted in 24 Spanish hospitals serving a population of 3.5 million people. The study included only patients who met criteria for definitive or probable CI. A website (www.colitisisquemica.org) provided logistical support., Results: A total of 364 patients met criteria for inclusion. CI was suspected clinically in only 24.2% of cases. The distribution of clinical patterns was as follows: reversible colopathy (26.1%), transient colitis (43.7%), gangrenous colitis (9.9%), fulminant pancolitis (2.5%), and chronic segmental colitis (17.9%). A total of 47 patients (12.9%) had an unfavorable outcome as defined by mortality and/or the need for surgery. Multivariate analysis identified the following signs as independent risk factors for an unfavorable outcome: abdominal pain without rectal bleeding [odds ratio (OR) 3.9; 95% confidence interval (CI) = 1.6-9.3], non-bloody diarrhoea (OR 10; 95% CI = 3.7-27.4), and peritoneal signs (OR 7.3; 95% CI = 2.7-19.6). Unfavorable outcomes also were more frequent in isolated right colon ischaemia (IRCI) compared with non-IRCI (40.9 vs. 10.3%, respectively; p < 0.0001). The overall mortality rate was 7.7%., Conclusions: The clinical presentation of CI is very heterogeneous, perhaps explaining why clinical suspicion of this disease is so low. The presence of IRCI, and occurrence of peritoneal signs or onset of CI as severe abdominal pain without bleeding, should alert the physician to a potentially unfavorable course.

Acute esophageal necrosis is a rare disorder, and its etiology is unknown, the mechanism of damage being usually multifactorial and secondary to ischemic compromise, acute gastric outlet obstruction, and malnutrition. Endoscopic findings show circumferential black discoloration of the distal esophagus with proximal extension ending sharply at the gastroesophageal junction, which is the most common presentation. Prognosis depends on comorbid illnesses. In this study we analyze all cases reported in a retrospective analysis over a 2-year period to define risk factors, clinical presentation, endoscopic features, histological appearance, treatment and outcome. Our department has recorded 7 cases from 6,003 endoscopies performed in the last 2 years. The finding of a "black esophagus" represented 0.11% of cases.

Heterozygous familial hypercholesterolemia affects one every 400 individuals, is caused by mutations in the LDL receptor gene and is associated with premature coronary artery disease. Nowadays, LDL cholesterol can be efficiently reduced with the new therapies to reduce blood lipids. We report a female patient who consulted in 1975, when she was 46 years old, for severe hypercholesterolemia. In 2003, a sample of leukocyte DNA was obtained and the uncommon 1705+1G>A mutation of the LDL receptor gene was detected. No mutations in the apolipoprotein B gene were found. The patient was treated successfully with simvastatin 80 mg/day and ezetimibe 10 mg/day and LDL cholesterol levels were reduced below 200 mg/dl.

Background and Aims: One of the problems analysing cause-effect relationships of growth and environmental factors is that a single factor could be correlated with other ones directly influencing growth. One attempt to understand tropical trees' growth cause-effect relationships is integrating research about anatomical, physiological and environmental factors that influence growth in order to develop mathematical models. The relevance is to understand the nature of the process of growth and to model this as a function of the environment., Methods: The relationships of Aphananthe monoica, Pleuranthodendron lindenii and Psychotria costivenia radial growth and phenology with environmental factors (local climate, vertical strata microclimate and physical and chemical soil variables) were evaluated from April 2000 to September 2001. The association among these groups of variables was determined by generalized canonical correlation analysis (GCCA), which considers the probable associations of three or more data groups and the selection of the most important variables for each data group., Key Results: The GCCA allowed determination of a general model of relationships among tree phenology and radial growth with climate, microclimate and soil factors. A strong influence of climate in phenology and radial growth existed. Leaf initiation and cambial activity periods were associated with maximum temperature and day length, and vascular tissue differentiation with soil moisture and rainfall. The analyses of individual species detected different relationships for the three species., Conclusions: The analyses of the individual species suggest that each one takes advantage in a different way of the environment in which they are growing, allowing them to coexist.

Characteristics developed by Cactaceae for adaptation to climates where water is limited include crassulacean acid metabolism (CAM), a thick cuticle, and spines and trichomes that intercept a proportion of solar radiation. A few studies consider morpho-anatomical and physiological characteristics of Cactaceae seedlings, which may help understand their establishment, growth, and eventual reproduction. In this study, photosynthetic metabolism (titratable protons) and morpho-anatomical features of Stenocereus beneckei seedlings were examined under limiting water conditions. Soil moisture treatments consisted of -0.03, -0.5, -1.5, and -3.0 MPa, and seedling samples were taken at 3 h intervals on one day at 7 and 9 months of age with three replicates per treatment. The results show irregular fluctuations in acidity concentrations during the first 6 and 7 months of age; at 9 months, an increase in titratable proton values was observed during the night, and it seems that soil moisture does not determine CAM expression. Seedlings from smaller seeds are less tolerant to water stress, they had poor growth in all treatments, and at -3.0 MPa after 3 months of drought none survived. Anatomical observations show collapsed cells associated with a high accumulation of calcium oxalate crystals and starch grains, as a response to water deficit. Titratable acidity concentration increased with seedling age, and CAM expression did not accelerate with soil water deficit.

Objective: To know the relationship between bone turnover markers and urinary calcium excretion with and without oral calcium load in postmenopausal and young women., Patients and Method: A cross-sectional study in 100 women (50 with normal menstrual cycle and 50 postmenopausal). After 5 days with a specific diet, urine of 24 hours, 2 hours in fasting, and 4 hours after oral calcium load was collected (the last urine collection was only done in 20 patients in each group. In these patients, an x-ray absorptiometry of distal forearm and vitamin D levels were evaluated too). Calcium, alkaline phosphatase, calcitonin, parathormone, N-telopeptides of type I collagen, and osteocalcin serum levels were measured in fasting. Creatinine and calcium were measured in urine to obtain the Ca/cr index., Results: Postmenopausal women had lower calcitonin levels while the Ca/cr index of 2 hours in fasting was higher than the eumenoreic women. We found significant correlation between Ca/cr index of 2 hours with age (R = .21; p = .04), parathormone (R = .21; p = .04), and calcitonin levels (R = -.29; p = .005). There was not correlation with the urinary calcium excretion including the 4 hour postload collection with the biochemical markers of bone turnover or mineral bone density of distal forearm., Conclusions: The urinary Ca/Cr index of 2 hours in fasting is related to parathormone levels and has a negative relationship with calcitonin levels in postmenopausal women. However, the urinary calcium excretion after oral calcium load was not related with the bone turnover grade.

We report the case of a 64-year-old man without hemorrhagic history experiencing epistaxis. The standard hemostasis assessment including prothrombin index, activated partial thromboplastin time (APTT) and platelet count found an isolated abnormal APTT (105 sec vs 33 sec). Therefore, coagulation factors were explored. An acquired factor VIII deficiency (5%) with anti-FVIII antibody (4.5 Bethesda unit.mL-1) was recognised. This anti-FVIII antibody was related to a Chlamydia pneumoniae pulmonary infection. Treatment consisted of: i) Four successive anterior packing and activated factor VII infusion (Novoseven); ii) steroids injection and bi-antibiotherapy. The time course of the epistaxis was favourable under treatment.

Excessive hyperbilirubinemia can cause irreversible neurological damage in the neonatal period. However, the complete understanding of the pathogenesis of unconjugated bilirubin (UCB) encephalopathy remains a matter of debate. This study investigates whether UCB inhibits the endocytosis of cationized ferritin (CF) by cultured rat astrocytes. The relationship between endocytosis and MTT reduction, as well as changes on tubulin and glial fibrillary acidic protein (GFAP) assembly, were also evaluated. Inhibition of endocytosis was complete in the presence of 171 microM UCB, while a marked decrease of CF labeling was noticed for 86 microM UCB. In addition, MTT reduction was inhibited by 60 to 76% as UCB concentrations changed from 17 to 171 microM, while alterations on both GFAP and microtubule morphology were only achieved by cell exposure to 171 microM UCB. These findings indicate that inhibition of CF endocytosis in rat cortical astrocytes by UCB is a concentration-dependent process that appears to be primarily related to a direct effect on the cell membrane and not to any alteration of cytoskeletal microtubules and intermediate filaments.

The molecular basis of bilirubin toxicity to nerve cell function is still unclear. Since astrocytes are the main transporters of synaptically released glutamate and impaired glutamate uptake results in neuronal death, we investigated the effect of unconjugated bilirubin (UCB) on [(3)H]glutamate uptake in cultured rat astrocytes and the role of bilirubin ionization on toxicity. Astrocytes were incubated for 5-15 min, with UCB concentrations from 17 to 342 microM and UCB/albumin molar ratios of 0.2-3.0, at pH 7.0, 7.4, and 8.0. Exposure of astrocytes for 15 min to 85.5 microM UCB and 28.5 microM albumin resulted in a 63.1% decrease of glutamate uptake (p < 0.01). Interestingly, the effect demonstrated to be correlated with the UCB/albumin molar ratio (r = -0.986, p < 0.01) and a significant decrease was observed for a UCB/albumin molar ratio as low as 0.8. Inhibition of glutamate transport was also pH-dependent as it occurred at 7.4 (p < 0.05) and 8.0 (p < 0.01), but not at 7.0, suggesting that the monoanionic species of UCB accounted for the inhibition. These findings indicate that UCB, and more precisely the monoanionic species, impairs a crucial function of astrocytes such as glutamate transport and support a potential role of astrocyte function in the pathogenesis of UCB-related brain damage (kernicterus)., (Copyright 1999 Academic Press.)

Objective: To communicate our results with primary angioplasty (PA) and stenting de novo in acute myocardial infarction., Material and Methods: 163 patients (pts) had clinical diagnosis of MI with an average onset time of symptoms of 2.5 hours. They were divided in two groups: 100 pts with PA using only balloon (group I) and 63 pts with primary stenting (group II) (63 pts with 85 stents). A high pressure technique (14-16 atm) was used in group II and all pts were on salicylic acid and ticlopidine. Both groups were compared., Results: Clinical characteristics, infarct location and infarct related-artery were similar, being left anterior descending the more frequently vessel involved. Clinical success rate was higher in group II (97% vs 87%) and complication rate was threefold higher in group I (9% vs 3%). A mean follow up of 57 months in group I showed a 21% clinical and/or angiographic restenosis rate, while the 12 months follow up in group II failed to show any case of restenosis at all., Conclusions: The use of stents in PA improves the results achieved with balloon procedures. There is a greater success rate, less complications and better survival patients who had the procedure while being in cardiogenic shock.

The aminopeptidase PepC is a cysteine peptidase isolated from lactic acid bacteria. Its structural and enzymatic properties closely resembles those of the bleomycin hydrolases, a group of cytoplasmic enzymes isolated from eukaryotes. Previous biochemical and structural data have shown that the C-terminal end of PepC partially occupies the active site cleft. In this work the substrate specificity of PepC was engineered by deletion of the four C-terminal residues. The mutant PepCDelta432-435 cleaved peptide substrates as an oligopeptidase while the aminopeptidase specificity was totally abolished. The substrate size dependency indicated that PepCDelta432-435 possesses an extended binding site able to accommodate four residues of the substrate on both sides of the cleaved bond. The activity of PepCDelta432-435 towards tryptic fragments of casein revealed a preference for peptides with hydrophobic amino acids at positions P2 and P3 and for Gly, Asn and Gln at position P1. PepCDelta432-435 was shown to be highly sensitive to the thiol peptidase inhibitors leupeptin or E64 which are inefficient towards the wild-type PepC. In conclusion, deletion of the four C-terminal residues in PepC produces a new enzyme with properties resembling those of an endopeptidase from the papain family.

The distribution of vasoactive intestinal peptide (VIP)- and calcitonin gene-related peptide (CGRP)-immunoreactive nerves and 125I-labeled VIP- and CGRP-binding sites was studied in the hamster seminal vesicle of 12-, 30- and 60-day-old animals. In addition, the general innervation of the seminal vesicle was examined using the general neuronal marker synaptophysin. Our results show that the densities of the overall (synaptophysin immunoreactive) and CGRP-immunoreactive innervation is constant during the post-natal development of the gland. However, a significant decrease in VIP-containing nerves is observed at the end of puberty. The autoradiographic study revealed that in 12-day-old animals, the epithelium presents VIP binding sites. However, in 30-day-old animals, VIP binding sites are observed in the epithelium of only a few clumps of acini. In 60-day-old animals, the gland is composed of acini with dilated lumina where VIP binding sites are not detected. In all groups studied the epithelium does not exhibit CGRP binding sites. The seminal vesicle muscle layer displays specific binding sites for both VIP and CGRP at all post-natal developmental times, but the density of VIP binding sites is higher in 12- than in 30- and 60-day-old animals. Our results, showing the presence of specific VIP and CGRP binding sites during the development of the hamster seminal vesicle, suggest that these neuropeptides may be involved in the growth and differentiation of the gland.

We characterized the 3272-26A-->G mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, creating an alternative acceptor splice site in intron 17a, that competes with the normal one, although we predict from consensus values, with lower efficiency. We analyzed five Cystic Fibrosis (CF) Portuguese patients with the 3272-26A-->G/F508del genotype. Besides clinical and haplotype characterization of those patients, we report here results from CFTR transcript analysis in nasal brushings from all five patients. RT-PCR analysis supports alternative splicing in all patients and carriers, but not in controls. By sequencing, we determined that the alternative transcript includes 25 nucleotides from intron 17a, which predictively cause frameshift and a premature stop codon. The use of this alternative splice site causes a reduction in the levels of normal transcripts from the allele with this mutation and, most probably, of normal protein as well. By immunocytochemistry of both epithelial primary cell cultures and slices from CF polyps, CFTR protein is detected at the cell membrane, with three different antibodies. Ussing chamber analysis of one nasal polyp shows a high sodium absorption, characteristic of CF. Altogether, the results suggest that the main defect caused by the 3272-26A-->G mutation is a reduction in normal CFTR transcripts and protein and therefore this mutation should be included in class V, according to Zielenski and Tsui., (Copyright 1999 Wiley-Liss, Inc.)

To obtain insight into the basic mechanisms controlling endocytosis, we tested the effects of different perfusates containing the cytological stain light green on endocytosis and ultrastructure of the vacuolar apparatus in renal proximal tubule cells. Rat proximal tubules were microinfused in vivo for 2 min in the presence or absence of light green with the following solutions: (A) perfusates containing inorganic salts and (B) perfusates with organic components or protein. In other experiments, the tubules were first microinfused for 2 min with 0.9% NaCl in the presence or absence of light green, then 15 min later further microinfused with or without light green using either group A or B solutions in order to either aggravate or reverse possible changes. All infused tubules were fixed after 5 min with 1% glutaraldehyde and examined by electron microscopy. In tubules microinfused without light green, the endocytic vacuolar apparatus in the apical cytoplasm showed a normal ultrastructure. However, microinfusion of solutions containing light green in either inorganic salts or a low concentration of protein caused significant changes in the apical endocytic apparatus. Large endocytic vacuoles were absent, and invaginations and small endocytic vacuoles were decreased in frequency. On the other hand, the amount of dense apical tubules was significantly increased, and in some cells dense apical tubules had transformed into a cisternalike network. These changes were aggravated in tubules which received a second microinfusion of NaCl and reversed in tubules that received a second infusion of protein. Furthermore, in the tubules microinfused with light green using perfusates containing organic components or protein, the apical cytoplasm of proximal tubule cells showed an essentially normal endocytic apparatus. The present study demonstrates that microinfusion of renal proximal tubules with light green disrupted normal endocytic membrane trafficking and recycling. These changes could be prevented or reversed by microinfusion of solutions containing protein or organic components.

PepCs isolated from lactic acid bacteria and bleomycin hydrolases of eukaryotic organisms are strict aminopeptidases which belong to the papain family of thiol peptidases. The structural basis of the enzymic specificity of the lactococcal PepC has been investigated by site-directed mutagenesis. The deletion of the C-terminal residue (Ala-435) abolished the aminopeptidase activity, whereas this deletion led to a new peptidase specificity. The enzymic properties of wild-type and mutant PepCs demonstrate that the terminal alpha-carboxy group plays a key role in the strict aminopeptidase activity.

The interaction of inorganic mercury with human and bovine milk proteins was studied. Gel filtration chromatography of skimmed milk and whey incubated with mercury showed that, in human milk, mercury was mainly bound to caseins, while a low proportion was bound to albumin. In bovine milk, mercury was associated with two protein fractions, caseins and beta-lactoglobulin. Furthermore, it was shown by electrophoresis that mercury induced the formation of dimers of beta-lactoglobulin. Thus, in both human and bovine milk, mercury possessed greater ability to interact with milk proteins than to the low-molecular-weight substances. However, the pattern of mercury distribution was different between the milk of these two species.

Background: The establishment of the nasopharyngeal flora was followed in Costa Rican children from birth to 1 year of age., Methods: Nasopharyngeal cultures were obtained at 1 (n = 413), 3 (n = 393), 6 (n = 376) and 12 months (n = 356) of age from children representative of the population in the Puriscal district. Weekly cultures were obtained from a subcohort of these children (n = 101). Mother-infant diads (n = 95) and preschool children (n = 208) attending day-care centers were also studied., Results: The estimated proportion of colonized children in the population differed markedly depending on the frequency of culture. Quarterly cultures showed a slow increase in carrier rates from 3.9% for Haemophilus influenzae, 3.1% for Streptococcus pneumoniae and 6.5% for Moraxella catarrhalis at 1 month of age to 10.1% carrying H. influenzae and 19.4% carrying S. pneumoniae by the end of the first year. By quarterly culture the proportion of children colonized at least once was 36% for S. pneumoniae, 26% for H. influenzae and 28% for M. catarrhalis. In contrast weekly sampling showed that 95 to 100% of the children were colonized at least once during the first year of life with H. influenzae, S. pneumoniae or M. catarrhalis. Nasopharyngeal carriage of H. influenzae, S. pneumoniae and M. catarrhalis was low in the mothers, and very few mother-infant pairs carried identical bacteria at the same time. In contrast carrier rates were high in the siblings attending day care (H. influenzae 27.9%, S. pneumoniae 39.4%, both organisms 26.6%). Infants with siblings had significantly higher bacterial carriage at all ages than infants without siblings., Conclusions: Quarterly nasopharyngeal cultures showed that Costa Rican infants acquire their nasopharyngeal flora at a rate comparable with that for infants in developed countries and that siblings are an important source of the bacteria. Weekly samplings showed that virtually all children were colonized at least once during the first year of life.

The importance of neuronal factors in the normal physiology of the seminal vesicles has been traditionally underestimated when compared to the trophic role of androgens. Immunohistochemical, autoradiographical and pharmacological experiments have, however, raised the possibility that neuropeptides, such as vasoactive intestinal polypeptide (VIP), neuropeptide tyrosine (NPY) and calcitonin gene-related peptide (CGRP), are necessary for full seminal vesicle function and development. These neuropeptides may be involved in the regulation of secretion, smooth muscle tone and blood flow. Furthermore, neuropeptides may have functional interactions with androgens affecting, probably, androgen receptor-dependent gene expression in these glands. It is now timely to focus attention on the biological relevance of neuropeptides in the seminal vesicles.

In the present work we have investigated the effects of medium- (15 days) and long-term (2 months) castration on vasoactive intestinal peptide (VIP)-immunoreactive nerve fibres and 125I-labelled VIP binding sites in the adult hamster seminal vesicle. The density of VIP- and synaptophysin (general neuronal marker)-containing nerve fibres was determined in immunofluorescently stained cryostat sections using a computerised image analysis system. The morphological analysis of 125I-VIP binding sites in seminal vesicle cryostat sections was performed by quantitative receptor autoradiography. Our results show that the densities of the overall (synaptophysin immunoreactive) and VIPergic innervation increase in both medium and long-term castrated animals. In absolute terms, the quantity of VIP- and synaptophysin- containing nerves is not altered in medium-term castrates, but decreases for synaptophysin in long-term castrates. Medium-term castration does not affect the density of 125I-VIP binding sites in the gland muscular coat, but a significant decrease is observed after long-term castration. In conclusion, our results indicate that whereas VIP nerves are apparently unaffected by castration, 125I-VIP binding sites in the muscular coat of hamster seminal vesicle are sensitive to androgen levels.

Our initial experience with intracoronary stenting without oral anticoagulant is presented. From december 1994 to december 1995 we implanted 58 stents in 43 patients (36 males). Indications were: retraction 1, restenosis 3, dissection 8, acute myocardial infarction 13 and de novo 33. All patients received ticlopidin, aspirin and diltiazem before the procedure, heparin and intracoronary nitroglycerin were given during the procedure, and ticlopidin and aspirin for the next six weeks. Non-compliant balloons at 14-16 atmospheres were used for stent implantation. The balloon/artery ratio was 1:1. Implant sites were: 29 in left anterior descendent, 17 in right coronary artery, 7 in circumflex and 5 in vein grafts. This procedure was successful in 40/43 (93%) patients. One patient died and 2 had acute occlusion. One patient underwent coronary artery bypass grafting and the other underwent a new dilatation with higher pressure. There were no subacute occlusions. During 140 days mean time follow up: 2 patients had angina (incomplete revascularization) and 27 patients with negative stress test. No myocardial infractions or deaths were present during the follow up period. Stent implantation with high pressure technique and without oral anticoagulant in coronary arteries greater than 3.0 mm is a safe and effective method with high rate success and very low acute complication rate.

The effect of some milk components on the cellular uptake of cadmium has been studied using a human intestinal cell line (Caco-2). Cadmium uptake by Caco-2 cells increased with the concentration of this metal in the culture medium, in a saturable way. These cells were exposed to different concentrations of cadmium and the synthesis of metallothionein was studied by a cadmium-saturation method. The levels of metallothionein increased with the cadmium concentration in the medium up to 20 microM of metal. Supplementation of the culture medium with 10% bovine milk caused a 25% decrease in the uptake of cadmium with respect to that internalized by the cells maintained in the culture medium alone. However, the uptake of cadmium from the medium supplemented with 10% human milk was similar to that with serum-free medium. beta-Lactoglobulin interacted with cadmium when studied by equilibrium dialysis, showing a stoichiometric binding constant of 5 x 10(4) l/mol. Interaction of lactoferrin with cadmium, however, was negligible. When Caco-2 cells were incubated in culture medium containing lactoferrin, cadmium uptake decreased with respect to that observed incubating the cells in a medium containing beta-lactoglobulin or in the free-protein medium. The inhibitory effect of lactoferrin on the uptake of cadmium might be due to a reduction of the cell surface charge, through its binding to the membrane.

Culture and immunomagnetic separation-polymerase chain reaction assays (IMS-PCR) were used to isolate and identify Shigella flexneri, S. dysenteriae type I and S. sonnei in faeces from 250 children up to 5 years and from their mothers (n = 143) selected at random from a large urban marginal community of Costa Rica. Children hospitalized because of severe diarrhoea (n = 110) were also studied. Only S. flexneri, mainly serotype 2a, and S. sonnei were found by culture. All specimens in which Shigella was cultured were also positive by the corresponding IMS-PCR. S. flexneri was isolated by culture from 1 (0.7%) mother and 4 (1.6%) community children. S. sonnei was found in 2 (0.8%) children. An additional 12 S. flexneri and four S. sonnei in the community children were found by IMS-PCR. In total, Shigella was cultured from 0.7% of mothers and 2.4% of children. By the IMS-PCR 2% of mothers and 8% of children were positive. S. flexneri was isolated by culture from 14 (12.7%) hospitalized children and S. sonnei from 1 (0.9%). An additional 11 S. flexneri and three S. sonnei were found by IMS-PCR. In total, Shigella was cultured from 13.6% of hospitalized children. By the IMS-PCR 26% of them were Shigella positive. Thus IMS-PCR was more than twice as effective in diagnosing shigellae as culture. Twelve (60%) Shigella positive community children were above 3-years-old and 25% of them were under one year. Seven (35%) of the Shigella positive children had dysenteric and 9 (45%) normal stools. Half of the Shigella infected community children had been weaned before the 3 months of age. By the age of 5 months, 90% of them were already weaned. Seventeen (59%) of the hospitalized Shigella positive children were under 1 year of age. The stools were watery or semiliquid in 13 (45%) and dysenteric in 12 (41%) of them. We conclude that shigellosis is common in Costa Rica and represents an important cause of severe infant diarrhoea requiring hospitalization.

The distribution of calcitonin gene-related peptide (CGRP)-immunoreactive nerves and CGRP binding sites, as well as the effect of CGRP on the muscle tension, was studied in the hamster seminal vesicle and coagulating gland. The use of an immunofluorescence staining technique on cryostat sections revealed that in the hamster seminal vesicle and coagulating gland, CGRP-positive nerve fibers are found in the connective interstitium and in the muscular and mucosal layers. Using an in vitro receptor autoradiographic technique, CGRP binding sites were found associated with the muscular coat. CGRP (10 pM to 1 microM) relaxed the seminal vesicle and the coagulating gland precontracted by either noradrenaline (10-30 microM) or the alpha 1-agonist, phenylephrine (10 microM). In preparations contracted by carbachol (10 microM), CGRP relaxed the seminal vesicle but not the coagulating gland. In both preparations, CGRP (1 microM) did not affect the muscle resting tension. These results suggest that CGRP may act as an inhibitory modulator of the autonomic control of contractility in the male accessory sex glands of the hamster.

The percentage of cadmium or lead present in the fat fraction of bovine milk is not affected by heating or freezing. In human skimmed milk, cadmium is mainly associated with a fraction with molecular weight lower than 10,000. Storage at -20°C for 10 days does not have any effect on the distribution of cadmium when milk is incubated with this metal before freezing. This treatment causes only a small increase in the amount of cadmium associated with the low molecular weight fraction when the metal is added after freezing. In bovine milk, 64% of cadmium is associated with a fraction with molecular weight above 70,000. Freezing causes a 37% decrease of the cadmium present in this fraction when the metal is added after thawing. When bovine milk was incubated with cadmium before freezing there was not a marked change in its distribution as when added after thawing. Heating at 63°C for 30 min caused a slight decrease in the amount of cadmium present in the casein fraction. The distribution change of cadmium after freezing or heating is probably due to the formation of complexes between the whey proteins and the metal, or to the disaggregation of the cadmium bound to casein micelles. Lead is mainly associated with caseins in bovine and human milk. No significant changes were caused by freezing or heating in the distribution of lead in human and bovine milk.

Primary cultures of epithelial cells from the hamster seminal vesicle were established in a chemically defined medium supplemented with hormones and growth factors. Epithelial cell clusters were prepared combining enzymatic dissociation and mechanical disaggregation and then seeded in bicameral systems equipped with collagen-membrane inserts. A growth curve was generated and the cells were characterized morphologically and morphometrically by light and electron microscopy. The immunocytochemical detection of cytokeratins and the measurement of transepithelial electrical resistance were also performed. The secretory activity was studied by fluorography using L-[35S]methionine as a precursor, and endocytosis was approached using horseradish peroxidase as a tracer. Our results show that epithelial cells of the seminal vesicle can be grown as a monolayer of morphological and functionally polarized cells which retain secretory and endocytic activities. These cell cultures might therefore prove useful to investigate further the regulation of secretion and endocytosis in the seminal vesicle and are a promising model to approach, in a broader scope, cell polarity and protein sorting and targeting.

Bradykinin (BK) and Tyr8-BK induced graded rat paw edema with EC50 values of 1.9 and 1.1 nmol/paw, while des-Arg9-BK (DABK, up to 300 nmol/paw) was marginally effective. Tyr8-BK, but not DABK, also caused a dose-related increase in mouse paw edema, with an EC50 of 1.3 nmol/paw. The response to Tyr8-BK (10 nmol/paw) in rat paw edema was antagonized by B2 receptor antagonists (HOE-140 or NPC 17731, 30 nmol/paw) but not by the B1 antagonist des-Arg9[Leu8]BK (DALBK, 100 nmol/paw). Daily intraplantar injections of Tyr8-BK (10 nmol/paw) for 7 days caused progressive desensitization (D) of edema in sham-operated and adrenalectomized Wistar rats. DABK (100 nmol/paw) caused marked paw edema in D paws from both groups, which was inhibited by DALBK (100 nmol/paw) and by dexamethasone (0.5mg/kg, s.c.). Systemic injection of lipopolysaccharide (10 micrograms/mouse, 24 h prior) potentiated the first and second phases of Formalin-induced pain but had no effect on paw edema. Coinjection of DABK (2-22 nmol/paw) with low doses of Formalin in lipopolysaccharide-treated mice, which had no effect on naive animals, dose dependently potentiated both phases of Formalin-induced pain but did not modify paw edema. These effects were antagonized by DALBK with ID50 values of 21.9 (first phase) and 64.1 (second phase) nmol/paw. Thus, both progressive desensitization of B2 receptors and systemic treatment with lipopolysaccharide induce a glucocorticoid-sensitive upregulation of B1 receptors mediating paw edema in the rat and Formalin-induced nociception in mice. These results suggest that induction of upregulation of B1 receptors may play important roles in controlling inflammatory processes and hyperalgesia.

Distribution of added lead and cadmium to bovine and human milk and whey has been studied. In bovine milk, about 97 and 89% of lead and cadmium, respectively, were recovered in the casein fraction obtained by enzymatic coagulation. However, only 6% of lead and 41% of cadmium were found in the same fraction separated by acid precipitation, indicating that the distribution of both metals is very different depending on the method used for milk fractionation. Moreover, gel filtration of bovine and human skimmed milk and whey after addition of lead and cadmium was carried out. Most of the lead was associated to the casein fraction after gel filtration of skimmed milk, whereas in the chromatography of whey, lead was eluted with the low molecular weight fraction in both species. However, a different pattern in the distribution of cadmium has been observed in the two species studied. In contrast to the binding of cadmium to the low molecular weight fraction in human skimmed milk and whey, it was mainly associated to a component of a molecular weight around 70,000 in bovine skimmed milk. This component was not present in bovine whey indicating that it is separated with casein during fractionation.

In newborn rodents, seminal vesicle epithelium (SVEP) cells display a poorly developed rough endoplasmic reticulum (RER) and Golgi complex, and they show no signs of secretion. From puberty onward, secretory material starts to appear, and the RER and Golgi complex progressively develop and reorganize until the adult ultrastructure is established around 40-60 days of age. Multivesicular bodies and lysosomes follow in this development but lysosomes evolve to lipofucsin granules with aging. The duration of the secretory cycle in SVEP cells is shorter than in other exocrine cells and the secretory protein pattern depends on the animal species, androgen status, and sexual activity. SVEP cells are also involved in endocytosis, which is coupled to exocytosis, and their endocytic pathway intersects the exocytic pathway in Golgi cisterns. The structure and function of SVEP cells depends mainly on testosterone, but other hormones and factors, such as the neuropeptide VIP, also influence their activity. Castration leads to programmed death and regression of SVEP cells to an extent that depends on the animal species. In addition, castration induces changes in the secretory protein pattern and delays its intracellular transport. Endocytic kinetics is also delayed following castration. Primary cultures of SVEP cells in a bicameral system are proposed as a model to investigate the activities of SVEP cells further.

Acquired immunodeficiency syndrome (AIDS) was first detected in Costa Rica in 1983. For four years most known cases were in hemophiliac men. Thereafter, AIDS in homosexual and bisexual men predominated. By December 31 of 1993, 563 persons had been diagnosed with the syndrome, 71% of them homosexual and bisexual men, 10% heterosexual men and women, 6% hemophiliacs, 2% intravenous drug abusers (IVDA's), 2% women and men who had blood transfusions, 1.4% infants born to HIV-infected mothers and 7% unknown. The epidemics in homosexual/bisexual men and in heterosexual women and men are rising; cases in infants and in persons who received blood or coagulation factors, are stagnant. The steady increase in AIDS among women is linked to exposure to bisexual partners. The moderate nature of the national epidemic reflects, in part, the low incidence of IVDA, the universal screening of blood donors for antibodies to the human immunodeficiency virus (HIV) since 1985, and the prompt banning of unsafe coagulation factors. The projection of AIDS for the year 2000 is 2,304 cases (606 accumulated incidence per million inhabitants). A national educational campaign, radio and television programs and other preventive actions, apparently did not influence the rate of receptive anal intercourse without condom (about 80%) during 9 years of the epidemic. Persons with HIV/AIDS often are deprived of social and medical benefits or are subjected to harassment and exploitation by the health sector. More efficient prevention must target children, adolescents and adults in reproductive age, to promote safer lifestyles, through education and counseling effected through primary health care.

Millions of Vibrio cholerae O1 El Tor were rapidly eliminated when added to commercial ceviche prepared by marination of mahi-mahi fish in lime juice. Likewise, large masses of viable vibrios present in laboratory contaminated fish, were readily eliminated after immersion in lime juice, during the preparation of ceviche. The killing effect was evident within 5 min of exposure of vibrios to lime juice, with reductions of more than 99.9% of the initial bacterial mass. After 2 h of marination of fish with lime juice (the minimum recommended), no vibrios were detected in the lowest working dilutions (1:10, 1:100). The Vibrio mass eliminated by lime juice was 2 to 6 logarithms greater than the maximum infectious dose, and 4 to 8 logs greater than the minimum infectious dose to induce cholera El Tor. Also, the killing time was shorter than the elapsing time between preparing and serving food in homes or restaurants. The traditional marination of fish with lime juice or its addition to seafood and meals immediately before consumption, should be protected and promoted to prevent infection with cholera vibrios. In the face of an epidemic of cholera, consumption of ceviche prepared with lime juice would be one of the safest ways to avoid infection with V. cholerae.

Lime juice killed millions of Vibrio cholerae O1, El Tor, Inaba, present on cabbage and lettuce contaminated in the laboratory. The lethal effect was evident within 5 min of exposure to lime juice. No vibrios could be recovered at dilution 1:10 using alkaline peptone water (APW) and thiosulfate-citrate-bile salts-saccharose agar (TCBS). More than 99.9% of the initial inoculum was effectively destroyed. The number of vibrios killed by lime juice was 2 to 6 logarithms greater than the maximum infecting dose, and 4 to 8 logs greater than the minimum infecting dose for cholera El Tor. The time interval needed for killing was smaller than the usual waiting time for serving food in homes and restaurants. The addition of lime juice to non-acidic foods, beverages and water, is strongly recommended to prevent infection with cholera vibrios and other acid-sensitive microorganisms. This measure is particularly important for rural and slum populations in the tropics and subtropics.

From October 1985 to May 1992, 400 patients had coronary angioplasty of 518 stenotic lesions with a 92% success rate. Mean stenosis was reduced from 88 to 15%. We had 25 major complications (6%): 8 emergency bypass surgery (2%), MI in 12 pts (3%) and 5 deaths (1%). Follow-up was possible in 329 of the 367 patients with a successful procedure (90%), with a mean follow-up of 44 months. We performed coronary angiogram in 113 patients with clinical data suggestive of restenosis, restenosis was found in 73, progression of the coronary atherosclerosis in 18 and 22 (19%) had no evidence of restenosis. A second coronary angioplasty was performed in 44 pts with a 93% success rate, 22 were sent to coronary bypass surgery and medical treatment was given to 7 patients. The 18 pts with progression of the disease were also sent to bypass surgery. During follow-up 25% of the patients required a second angioplasty or bypass surgery. A total of 115 (35%) pts showed evidence of restenosis: in 73 pts (22%) a coronary angiogram revealed restenosis, 12 had angina, 15 myocardial infarction and 15 died of cardiovascular disease. At the end of follow-up 259 pts (79%) were free of myocardial infarction, bypass surgery or cardiovascular death. Survival rate at 3.7 years was 94.5%. In conclusion, coronary angioplasty showed to be safe, effective and with a good long term outcome.

The presence and functional role of vasoactive intestinal peptide in the hamster seminal vesicle were studied by a combination of structural and functional approaches. The use of an immunofluorescence staining technique in both cryostat sections and whole-mount preparations revealed that vasoactive intestinal peptide-immunoreactive nerve fibres were mainly localized in the lamina propria of the mucosal layer. In double-stained preparations, vasoactive intestinal peptide immunoreactivity was found to be localized in nerves also containing acetylcholinesterase activity. At the ultrastructural level, the use of an immunogold staining method showed that vasoactive intestinal peptide immunoreactivity occurred in large granular vesicles (80-150 nm in diameter) in nerve varicosities which also contained small pleomorphic agranular vesicles. In order to evaluate the anatomical distribution of vasoactive intestinal peptide binding sites in the seminal vesicle, we have utilized an in vitro receptor autoradiographic technique. Vasoactive intestinal peptide binding sites were localized in the basal region of the secretory epithelium, in the muscle layer and in the wall of blood vessels. In vitro incorporation of [3H]L-leucine into protein by tissue slices revealed that vasoactive intestinal peptide (1 microM) significantly increases the amount of released protein. Vasoactive intestinal peptide (0.1-1 microM) did not affect the resting tension of the muscle but significantly inhibited the increase in muscle tension induced by carbachol. Atropine prevented the effect of carbachol, indicating that the latter is mediated by muscarinic receptors. Our results suggest that in the hamster seminal vesicle, vasoactive intestinal peptide is involved in the modulation of muscarinic function and in the control of secretion.