RÉSUMÉ
For some infrastructures such as oil and gas extraction boreholes or radioactive waste repositories, where distributed optical fiber sensors are employed to grant the safety of the facilities, the presence of gas species such as hydrogen or deuterium is one of the most relevant parameters to monitor. The possibility of employing the same kind of sensors for this purpose is of special interest, reducing the cost by employing a single interrogator, able to measure multiple parameters by simply employing adequate sensing fibers. To meet this goal, we present here a chemical sensor based on chirped-pulse phase-sensitive optical time-domain reflectometry (CP-φOTDR), which is able to detect these species while they diffuse into the silica fiber. The ability of chirped-pulse φOTDR to measure a change in refractive index with sensitivity around 10-8 has allowed determining hydrogen concentration with accuracy on the order of 10-3 mol/m3 and spatial resolution â¼6 m. Another experiment provides an indirect measurement of the solubility of deuterium in a standard telecom-grade optical fiber, which is found to be around 1.47×1024 m3/bar.
RÉSUMÉ
Optical fibers hold promise for accurate dosimetry in small field proton therapy due to their superior spatial resolution and the lack of significant Cerenkov contamination in proton beams. One known drawback for most scintillation detectors is signal quenching in areas of high linear energy transfer, as is the case in the Bragg peak region of a proton beam. In this study, we investigated the potential of innovative optical fiber bulk materials using the sol-gel technique for dosimetry in proton therapy. This type of glass is made of amorphous silica (SiO[Formula: see text]) and is doped with Gd[Formula: see text] ions and possesses very interesting light emission properties with a luminescence band around 314 nm when exposed to protons. The fibers were manufactured at the University of Lille and tested at the TRIUMF Proton Therapy facility with 8.2-62.9 MeV protons and 2-6 nA of extracted beam current. Dose-rate dependence and quenching were measured and compared to other silica-based fibers also made by sol-gel techniques and doped with Ce[Formula: see text] and Cu[Formula: see text]. The three fibers present strong luminescence in the UV (Gd) or visible (Cu,Ce) under irradiation, with the emission intensities related directly to the proton flux. In addition, the 0.5 mm diameter Gd[Formula: see text]-doped fiber shows superior resolution of the Bragg peak, indicating significantly reduced quenching in comparison to the Ce[Formula: see text] and Cu[Formula: see text] fibers with a Birks' constant, k[Formula: see text], of (0.0162 [Formula: see text] 0.0003) cm/MeV in comparison to (0.0333 [Formula: see text] 0.0006) cm/MeV and (0.0352 [Formula: see text] 0.0003) cm/MeV, respectively. To our knowledge, this is the first report of such an interesting k[Formula: see text] for a silica-based optical fiber material, showing clearly that this fiber presents lower quenching than common plastic scintillators. This result demonstrates the high potential of this inorganic fiber material for proton therapy dosimetry.
Sujet(s)
Gadolinium , Fibres optiques , Protonthérapie , Radiométrie/instrumentation , Humains , Transfert linéique d'énergie , Luminescence , Fantômes en imagerie , Protons , Radiométrie/méthodes , SiliceRÉSUMÉ
We report a method for fabricating fiber Bragg gratings (FBG) resistant to very severe environments mixing high radiation doses (up to 3 MGy) and high temperatures (up to 230°C). Such FBGs have been written in two types of radiation resistant optical fibers (pure-silica and fluorine-doped cores) by exposures to a 800 nm femtosecond IR laser at power exceeding 500 mW and then subjected to a thermal annealing treatment of 15 min at 750°C. Under radiation, our study reveals that the radiation induced Bragg wavelength shift (BWS) at a 3 MGy dose is strongly reduced compared to responses of FBGs written with nonoptimized conditions. The BWS remains lower than 10 pm for temperatures of irradiation ranging from 25°C to 230°C without noticeable decrease of the FBG peak amplitude. For an applicative point of view, this radiation induced BWS corresponds to an additional error on the temperature measurements lower than 1.5°C, opening the way to the development of radiation-tolerant multi-point temperature sensors for nuclear industry.
RÉSUMÉ
A novel cellulase was isolated from the thermoacidophilic bacterium Alicyclobacillus acidocaldarius ATCC27009 grown in medium containing carboxymethylcellulose. The enzyme is a glycosylated monomer of 56.2 kDa, relatively thermostable, with optimal pH and temperature of 4.0 and 65 degrees C, respectively. Enzymatic assays on several polysaccharides demonstrated that CelG was specific for carboxymethylcellulose.
Sujet(s)
Cellulase/composition chimique , Cellulase/isolement et purification , Bactéries à Gram positif formant des endospores/enzymologie , Cellulase/métabolisme , Chromatographie d'affinité , Chromatographie sur gel , Électrophorèse sur gel de polyacrylamide , Concentration en ions d'hydrogène , Spécificité du substrat , Température , UltracentrifugationRÉSUMÉ
AIMS: It is evaluated the effectiveness of the combined action of two highly thermostable enzymes for the hydrolysis of xylans at high temperature in order to produce D-xylose. METHODS AND RESULTS: Xylans from different sources were hydrolyzed at high degree at 70 degrees C by co-action of a xylanase from the thermophilic bacterium Anoxybacillus flavithermus BC and the novel beta-xylosidase/alpha-arabinosidase from the hyperthermophilic crenarchaeon Sulfolobus solfataricus Oalpha. Beechwood xylan was the best substrate among the xylans tested giving, by incubation only with xylanase, 32.8 % hydrolysis after 4 h. The addition of the beta-xylosidase/alpha-arabinosidase significantly improved the rate of hydrolysis, yielding 63.6% conversion after 4 h incubation, and the main sugar identified was xylose. CONCLUSIONS: This study demonstrates that a significant degree of xylan degradation was reached at high temperature by co-action of the two enzymes. Xylose was obtained as a final product in considerable yield. SIGNIFICANCE AND IMPACT OF THE STUDY: Although the xylan represents the second most abundant polysaccharide in nature, it still doesn't have significant utilization for the difficulties encountered in its hydrolysis. Its successful hydrolysis to xylose in only one stage process could make of it a cheap sugar source and could have an enormous economic potential for the conversion of plant biomass into fuels and chemicals.
Sujet(s)
Bacillaceae/enzymologie , Endo-1,4-beta xylanases/métabolisme , Sulfolobus solfataricus/enzymologie , Xylanes/métabolisme , Bacillaceae/croissance et développement , Chromatographie en phase liquide à haute performance/méthodes , Diholoside/analyse , Glucosidases/métabolisme , Glycosidases/métabolisme , Température élevée , Hydrolyse , Triholosides/analyse , Xylanes/analyse , Xylose/analyseRÉSUMÉ
Diagnostic surveys carried out on ovine larval echinococcosis in slaughtering plants at a national level in 1998 made it possible to meet several objectives, i.e. to estimate the existing levels of prevalence, to evaluate the effectiveness of the control actions applied by the Programme against Hydatidosis and to contribute to the improvement of the health information systems for epidemiological surveillance. The work included sheep (Ovis aries), both lambs (2-4 teeth) and adults (6-8 teeth). A random sample was performed among sheep from the plants that slaughter the majority of animals in the country. The sample included 22 animals per herd, and it involved removing their offal (liver, lungs and heart) and processing serial sections at the laboratory. The histopathological studies were performed on the lesions that could not be adequately identified at gross examination, and those that were so small that had to be submitted for confirmation of the diagnosis. Of the 2035 animals that were examined, 1019 were lambs (2-4 teeth) and 1016 were adults (6-8 teeth). The prevalence of larval echinococcosis was 7.7% in lambs and 18.0% in adults. The overall prevalence in the liver was 8.5 and 8.0% in the lungs. As to the characteristics of the morphologic structures of the larvae, in younger animals 29% of the lesions were calcified, and 71.0% were hyaline, while in older animals the proportions were 34.4% calcified versus 65.6% hyaline. Other conditions found in the liver parenchyma included: white stain (0.8%), caseous lymphadenitis (3.0%), abscess (0.4%), larvae of Taenia hydatigena (5.1%), Fasciola hepatica (3.9%). And 1.7% of the animals presented larvae of Taenia ovis in the cardiac muscle. Previous surveys carried out in 1994 using the same methodology showed a prevalence of 15.2% in 2-4 teethed animals and 33.9% in 6-8 teethed animals, thus confirming that rates have decreased by 49.6 and 47.0%, respectively. The data obtained reflect the reduction of areas contaminated with eggs of Echinococcus granulosus, which would cause a loss in immunity against this parasite after a certain time. Prevalence of larvae T. hydatigena and T. ovis could change if more sites were considered, a fact that was confirmed in studies carried out in our country. The species O. aries must be used as an indicator in the control programmes, especially in the category of 2-4 teethed lambs. The fact they are not as frequent in our market must be considered; in 1998 they accounted for only 3.0% of the total numbers of animals.
Sujet(s)
Abattoirs , Échinococcose/médecine vétérinaire , Echinococcus/isolement et purification , Maladies des ovins/épidémiologie , Ovis aries/parasitologie , Facteurs âges , Animaux , Échinococcose/épidémiologie , Échinococcose/prévention et contrôle , Echinococcus/classification , Echinococcus/croissance et développement , Fasciola hepatica/croissance et développement , Recommandations comme sujet , Étapes du cycle de vie , Modèles logistiques , Prévalence , Maladies des ovins/prévention et contrôle , Taenia/classification , Taenia/croissance et développement , Uruguay/épidémiologieRÉSUMÉ
This study attempted control of transmission of Echinococcus granulosus from dogs to sheep in different areas in the Department of Florida, Uruguay, by treating dogs with praziquantel at intervals of 6, 12 and 16 weeks. The 6-week interval was based on the prepatent period of infection with E. granulosus, the 12- and 16-week intervals were based on the rate of reinfection with tapeworms in dogs in the area. Dogs had become reinfected with E. granulosus between 2 and 4 months after treatment, whereas they became reinfected with the Taenia spp. tapeworms within 2 months of treatment. One year after the start of treatments sentinel lambs were born and grazed the farms in the three treatment areas. Approximately, 15 months later when the sentinel lambs were killed and examined for parasites the six weekly treatments had stopped the transmission of E. granulosus to the sentinel lambs. Treatment of dogs at 12- and 16-week intervals failed to stop transmission of E. granulosus but both the numbers of farms and the numbers of sheep infected with E. granulosus were lower where dogs received 12 weekly treatments compared with dogs receiving 16 weekly treatments and a fourth area where dogs had received no treatments (chi(2)P=0.002). Lambs continued to become infected with the Taenia spp. tapeworms in all the areas. Control was complicated by large changes in the dog population. From a starting population of 1164 dogs in the three treated areas, 832 new dogs, most of these adult hunting dogs, entered the population and 793 dogs were lost from the population.
Sujet(s)
Anthelminthiques/usage thérapeutique , Maladies des chiens/prévention et contrôle , Échinococcose/médecine vétérinaire , Praziquantel/usage thérapeutique , Maladies des ovins/prévention et contrôle , Animaux , Maladies des chiens/traitement médicamenteux , Maladies des chiens/transmission , Chiens , Échinococcose/traitement médicamenteux , Échinococcose/transmission , Echinococcus/effets des médicaments et des substances chimiques , Echinococcus/croissance et développement , Ovis , Maladies des ovins/transmission , Facteurs temps , UruguayRÉSUMÉ
A pharmacokinetic study based on the distribution of radioactivity from the double labelled S-adenosyl-L-methionine (SAM) has been carried out by oral administration of the liposoluble stable salt [methyl-(14)C, 8-(3)H]SAM N-ole-1-oyltaurate to rats. The SAM sulfate p-toluensulfonate salt, the only SAM salt at present commercialized as drug, was chosen as reference compound to have a comparative pharmacokinetic analysis. The metabolism of the SAM is characterised by a differential use of the two labelled moieties by the various organs, liver being the most active in metabolizing the sulfonium compound with a preferential uptake of the methyl-(14)C fragment. The radioactivity detected after the administration of [methyl-(14)C, 8-(3)H]SAM N-ole-1-oyltaurate is, in all the organs examined, two times higher than the [methyl-(14)C, 8-(3)H]SAM sulfate p-toluensulfonate compound, attesting that the liposoluble [methyl-(14)C, 8-(3)H]SAM N-ole-1-oyltaurate is provided with better bioavailability.
Sujet(s)
Acides oléiques/pharmacocinétique , Adémétionine/analogues et dérivés , Adémétionine/pharmacocinétique , Taurine/pharmacocinétique , Administration par voie orale , Animaux , Biodisponibilité , Chromatographie en phase liquide à haute performance , Fèces/composition chimique , Foie/composition chimique , Foie/métabolisme , Mâle , Acides oléiques/administration et posologie , Rats , Rat Wistar , Adémétionine/administration et posologie , Taurine/administration et posologie , Taurine/analogues et dérivés , Distribution tissulaireRÉSUMÉ
The aim of the experiment was to determine whether supplementation of the lamb-ewe unit during intra-uterine and postnatal life affects testicular stereology, particularly Sertoli cell numbers, in 120 pregnant Corriedale ewes grazed either native pastures (control group) or improved pastures+grain supplement (treated group). Ewes bearing single ram lambs were maintained under the same feeding regime until lambs were castrated (99 days of age). Body weight, testosterone and FSH blood serum levels were recorded at 45, 75 and 99 days of age. Body weight was higher (P<0.01) in the treated group from birth on. Serum testosterone values did not differ between groups. Serum FSH values tended to differ at 45 days of age (P<0.06). Testicular weight and testes histology showed earlier pubertal development and a tendency for higher Sertoli cell numbers in the treated (supplemented) group. This tendency may indicate that extensively reared lambs supplemented during fetal and postnatal life have higher testicular growth and sperm production in later life.
RÉSUMÉ
Both "environmental chemistry" and metabolic biochemical reactions can constantly generate in vivo free radicals and other oxygen-derived species that can cause severe damage to almost all biomolecules, especially to DNA, proteins, and lipids. The superoxide anion has been shown to be the most readily generated and spread radical among organisms and it is a common intermediate of oxidative stress processes in the cells. The antioxidant defense system of superoxide dismutases (SOD) scavenges and minimizes the formation of this radical, and thus plays a major role in reducing cumulative oxidative damage in different cell compartments both in aerobic and anaerobic cells. In the cell, cytosol SODs are constitutively present and induced by many oxidative agents able to raise the superoxide concentrations. Presence of SODs, however, in extracellular cell-associated locations demonstrates how valuable they are in maintaining the integrity of cells against oxidative stress generated by the cell environment, particularly upon increased oxygenation. Because SODs have recently been found in Archaea, which are prokaryotes, sometimes living in extreme environments, even in anaerobic ones, these enzymes can be considered essential: they may have allowed the evolution of aerobic respiration starting from an ancient form of oxygen-insensitive life.
Sujet(s)
Archéobactéries/métabolisme , Stress oxydatif , Oxygène/métabolisme , Superoxide dismutase/métabolismeRÉSUMÉ
The aim of the present study was to determine whether pre- and post-pubertal young rams on different grazing regimes, resulting in differences in live weight (LW), would show corresponding differences in testicular growth or testicular morphometry that could influence the reproductive traits of these rams upon reaching adulthood. Forty-one spring-born Corriedale rams were reared on either native pasture (low feeding level, Group L, n=22) or improved pasture (higher feeding level, Group H, n=19) from 1 to 7 months of age. Thereafter, half the animals in the native-pasture group were placed on improved pasture and vice versa, thus creating an additional four differential-grazing treatment groups (Groups LL, n=11; LH, n=11; HL, n=10; and HH, n=9). Animals were managed in this way until 18 months of age. Half the animals from each group were then castrated and their testes were subjected to morphometric analysis. The remaining animals (Groups LL, n=6; LH, n=6; HL, n=5; and HH, n=4) were managed together until 30 months of age (from 18 to 27 months on native pastures and from 27 to 30 months of age on improved pastures, at a stocking rate of two to three rams per hectare), whereupon they were also castrated for testicular morphometry. LW and scrotal circumference (SC) were recorded every 60 days. The stereological analysis of testicular parenchyma included counts of elongated spermatids and Sertoli cells. Differences (P<0.001) in LW were observed between feeding levels, even at 30 months of age. Differences (P<0.001) in SC existing at the end of the differential treatment (18 months of age) disappeared (n.s.) soon after. Most differences (P<0.05) in testicular morphometry existing at the end of the differential treatments were no longer significant 1 year later. It is concluded that changes in grazing management during pre- and post-pubertal periods can induce short-lived differences in testicular post-natal growth in Corriedale rams but do not influence testicular morphology or function later in life.
Sujet(s)
Aliment pour animaux , Élevage , Ovis/croissance et développement , Testicule/croissance et développement , Animaux , Poids , Castration/médecine vétérinaire , Traitement d'image par ordinateur , Mâle , Répartition aléatoire , Scrotum/physiologie , Cellules de Sertoli , Ovis/physiologie , Spermatides/physiologie , Statistique non paramétrique , Testicule/anatomie et histologie , UruguayRÉSUMÉ
S-adenosyl-L-methionine (SAM) is an important metabolic intermediate that serves as a donor of methyl and aminopropyl groups to a variety of acceptor molecules. The molecule in vitro is unstable both in solution and in crystalline form undergoing irreversible conversion to 5'-methyltioadenosine (MTA) and homoserine lactone. Since this form of instability seems to be prevented in the cell of the living organism by bonds with macromolecules, we designed and developed a novel class of salts of SAM with large size anions to improve the stability of the sulfonium compound outside the cell. For this purpose we synthesised and characterised by NMR and IR spectroscopy anions consisting of amidic derivatives of taurine with fatty acids. Stability studies performed with the new SAM salts indicate that SAM becomes much more stable when it interacts with large size anions and in fact, more than 84% of the SAM is recovered after 36 months in lyophilized samples. The high stability of the new products widens the possibility of new therapeutic applications of SAM in human therapy.
Sujet(s)
Adémétionine/analogues et dérivés , Taurine/analogues et dérivés , Anions , Stabilité de médicament , Lyophilisation , Concentration en ions d'hydrogène , Liposomes , Résonance magnétique nucléaire biomoléculaire , Adémétionine/composition chimique , Spectrophotométrie IR , Taurine/composition chimiqueRÉSUMÉ
The present study was conducted: (a) to determine the degree of seasonal variation in testis stereology in Corriedale rams between autumn and winter; (b) to test the hypothesis that testis stereology of Corriedale rams grazing native pastures during autumn and winter would differ from those of Corriedale rams grazing sown pastures and supplemented with grain during the same period; and (c) to determine whether Sertoli cell numbers differ in adult rams between the breeding season (autumn) and the following non-breeding season (winter). Twenty experimental animals were studied. Six rams (autumn control group, C-A) that had been grazing on native pasture (stocking rate = 2-3 animals ha(-1)) were castrated at the beginning of the experiment (March, early autumn). Seven rams (winter control group, C-W) continued to graze on native pasture at the same stocking rate until the end of the experiment (August, late winter). Another seven rams (treated group, T) grazed on improved pasture (stocking rate = 1-2 animals ha(-1)) and were supplemented with 1 kg grain ram(-1) day(-1) until the end of the experiment. Live weight, scrotal circumference, serum testosterone concentration and selected testicular stereological parameters were measured. The treatment did not impede the winter reduction in testicular activity and reduced its magnitude slightly (group T) compared with controls (group C-W). Sertoli cell numbers were higher in autumn (group C-A) than in winter, both on native (group C-W) and sown pastures (group T). Diminishing Sertoli cell numbers between autumn and the following winter suggest the occurrence of that Sertoli cell death during this period. The results indicate that, although the reproductive activity of Corriedale rams is moderately seasonal, a restricted change in grazing and grain supplementation can only modify it to a limited extent.
Sujet(s)
Élevage , Ovis/physiologie , Testicule/anatomie et histologie , Phénomènes physiologiques nutritionnels chez l'animal , Animaux , Castration/médecine vétérinaire , Traitement d'image par ordinateur , Mâle , Photopériode , Dosage radioimmunologique/médecine vétérinaire , Répartition aléatoire , Analyse de régression , Saisons , Cellules de Sertoli/physiologie , Cellules de Sertoli/ultrastructure , Ovis/anatomie et histologie , Numération des spermatozoïdes/médecine vétérinaire , Spermatides/physiologie , Testostérone/sang , UruguayRÉSUMÉ
The catalytic properties of a beta-galactosidase from Aspergillus oryzae, entrapped into a spongy polyvinyl alcohol cryogel, were studied. This polymeric matrix was selected because of its mild conditions of preparation and its stability, biocompatibility, structural strength and diffusive properties. The enzyme was entrapped, in high percentage, into cryogel sponges and its activity and kinetic parameters were determined and compared with those of the free enzyme, using as substrates o-nitrophenyl-beta-galactopyranoside (ONPG) or lactose. The immobilized enzyme showed a reduced activity with ONPG and lactose, probably because of substrate diffusion limitations through the matrix, but it was more stable to temperature, pH and ionic strength than the free enzyme. Lactose hydrolysis under continuous experimental conditions was performed using the matrix-enzyme cited above.
Sujet(s)
Aspergillus oryzae/enzymologie , Enzymes immobilisées , beta-Galactosidase/métabolisme , Animaux , Catalyse , Stabilité enzymatique , Température élevée , Concentration en ions d'hydrogène , Hydrolyse , Cinétique , Lactose/métabolisme , Nitrophénylgalactoside/métabolisme , Concentration osmolaire , Poly(alcool vinylique)RÉSUMÉ
Enzymes that convert starch and dextrins to alpha,alpha-trehalose and glucose were found in cell homogenates of the hyperthermophilic acidophilic archaeon Sulfolobus shibatae DMS 5389. Three enzymes were purified and characterized. The first, the S. shibatae trehalosyl dextrin-forming enzyme (SsTDFE), transformed starch and dextrins to the corresponding trehalosyl derivatives with an intramolecular transglycosylation process that converted the glucosidic linkage at the reducing end from alpha-1,4 to alpha-1,1. The second, the S. shibatae trehalose-forming enzyme (SsTFE), hydrolyzed the alpha-1,4 linkage adjacent to the alpha-1,1 bond of trehalosyl dextrins, forming trehalose and lower molecular weight dextrins. These two enzymes had molecular masses of 80 kDa and 65 kDa, respectively, and showed the highest activities at pH 4.5. The apparent optimal temperature for activity was 70 degrees C for SsTDFE and 85 degrees C for SsTFE. The third enzyme identified was an alpha-glycosidase (Ss alpha Gly), which catalyzed the hydrolysis of the alpha-1,4 glucosidic linkages in starch and dextrins, releasing glucose in a stepwise manner from the nonreducing end of the polysaccharide chain. The enzyme had a molecular mass of 313 kDa and showed the highest activity at pH 5.5 and at 85 degrees C.
Sujet(s)
Glucose/biosynthèse , Amidon/métabolisme , Sulfolobus/enzymologie , Tréhalose/biosynthèse , Glucosyltransferases/métabolismeRÉSUMÉ
A protein with beta-glycosidase activity from Sulfolobus solfataricus (S beta gly) was expressed in the yeast Saccharomyces cerevisiae. The purification procedure was made fast and easy by employing a single chromatographic step. After 5.8-fold purification, the cell extract gave a homogeneous enzyme at 166 U/mg. The recombinant enzyme was functionally and structurally similar to the wild-type enzyme. Kinetic experiments showed the same wide substrate specificity; in fact, the expressed enzyme hydrolyzed beta-D-gluco-, fuco-, and galactosides and a large number of glucoside dimers and oligomers, linked beta 1 -> 4. Moreover, the molecular mass of the enzyme was estimated to be 60 kDa by SDS-PAGE and 240 kDa by gel filtration, glycerol gradient, and ultracentrifugation analyses, indicating that the enzyme has a tetrameric structure. The N-terminal amino acid sequence, the temperature dependent activity, and content of secondary structure were similar to those of the wild-type enzyme. CD spectral and kinetic analyses showed that the only differences from the wild-type enzyme consist of the absence of lysine methylation, the presence of some glycosylated amino acid residues, and lower thermostability. Furthermore, calorimetric analyses on the expressed protein indicated values of delta dH = 5072 kJ/ mol and delta (d)C(p)= 100 kJ/mol, appreciably lower than those of the wild-type protein.
Sujet(s)
Saccharomyces cerevisiae/enzymologie , bêta-Glucosidase/composition chimique , Séquence d'acides aminés , Dichroïsme circulaire , Stabilité enzymatique , Glucosides/pharmacologie , Glycosylation , Concentration en ions d'hydrogène , Cinétique , Données de séquences moléculaires , Masse moléculaire , Protéines recombinantes/composition chimique , Saccharomyces cerevisiae/génétique , Sulfolobus/enzymologie , Sulfolobus/génétique , Température , bêta-Glucosidase/biosynthèse , bêta-Glucosidase/isolement et purificationRÉSUMÉ
The application of enzymes isolated from extreme thermophiles in biotechnological processes is hampered by their unconventional fermentation conditions. The expression, in mesophilic hosts, of genes encoding for thermophilic proteins enables these difficulties to be overcome and permits the production of enzymes in high yield by using conventional fermentation plants and an efficient enzyme purification utilizing heat precipitation of host proteins. The beta-glycosidase gene from Sulfolobus solfataricus, a thermoacidophilic archaeon growing at 87 degrees C and pH 3.5, has been cloned and expressed in Saccharomyces cerevisiae (baker's yeast). The fermentation of a S. cerevisiae strain on a 100-litre scale and the two-step purification of the expressed beta-glycosidase by cell autolysis and extracts thermal precipitation is described. This procedure, after 72 h of autolysis, gave a yield 56-fold higher with respect to that obtained with the beta-glycosidase from S. solfataricus.
Sujet(s)
Saccharomyces cerevisiae/enzymologie , bêta-Glucosidase/biosynthèse , Biotechnologie , Clonage moléculaire , Fermentation , Régulation de l'expression des gènes codant pour des enzymes/génétique , Concentration en ions d'hydrogène , Dénaturation des protéines , Sulfolobus/enzymologie , Sulfolobus/génétique , Température , bêta-Glucosidase/génétique , bêta-Glucosidase/isolement et purificationRÉSUMÉ
A base-line survey was carried out on the transmission dynamics of Echinococcus granulosus, Taenia hydatigena and Taenia ovis in sheep in the Department of Florida, Uruguay. Mean life expectancy at birth of the sheep population in this Department was estimated at 3.5 and 4.8 years for male and female sheep, respectively. Both intensity and prevalence of E. granulosus infection increased with age providing no evidence that immunity was acquired in the population. This revealed that E. granulosus was only relatively stable and in an equilibrium endemic steady state with an estimate of the basic reproduction ratio (Ro) of about 1.2. The mature larval population in sheep was over-dispersed with parameter k, from fitting a negative binomial, being 0.199 and 0.556 in 42- and 54-month-old sheep, respectively. The mean number of cysts increased by 1.05 cysts per year and the infection pressure was calculated as 0.174 infections per year, implying that each infection consisted of 693.6 eggs to produce 6.03 cysts. Neither intensity nor prevalence of infection with T. hydatigena or T. ovis increased with age. This indicated that immunity was acquired to reinfection by both parasites and they were in an equilibrium hyperenedemic steady state with an estimated Ro of 4.2-5.8. Viability and fertility of E. granulosus, and viability of T. hydatigena cysts in the older age groups were low. The effects of the different transmission levels of the parasites on potential control strategies are discussed.
Sujet(s)
Échinococcose/médecine vétérinaire , Maladies des ovins/transmission , Taeniase/médecine vétérinaire , Animaux , Chiens , Échinococcose/épidémiologie , Échinococcose/transmission , Echinococcus , Femelle , Humains , Mâle , Ovis , Maladies des ovins/épidémiologie , Maladies des ovins/prévention et contrôle , Taenia , Taeniase/épidémiologie , Taeniase/transmission , Uruguay/épidémiologieRÉSUMÉ
The present study was undertaken to estimate the time for the attainment of spermatogenesis in springborn Corriedale ram lambs under conditions of extensive grazing systems in Uruguay. Clinical (live weight, scrotal circumference, penile/preputial separation), morphological (light and electron microscopy) and endocrinological (testosterone levels) parameters were examined. Two experiments in 2 consecutive years were carried out. In Exp. I, 40 ram lambs were clinically examined, weighed, and blood-sampled at 2 week-intervals between 78 and 216 days of age. Sixteen were castrated in 3 selected periods: 132-162 (n:2), 145-175 (n:6) and 186-216 days (n:8). In Exp. II, 40 ram lambs appertaining to the next generation of the same flock were examined as above at 180-210 days of age, and castrated for morphological studies. The time for attainment of complete spermatogenesis correlated significantly with most corporal parameters (i.e. scrotal circumference (r = 0.52); testicular weight (r = 0.61), epididymal weight (r = 0.60), penile/preputial separation (r = 0.75). The age of castrated ram lambs correlated with the degree of spermatogenesis (r = 0.83), although no significant correlations were found with live weight or with levels of circulating testosterone. The histology showed major variations in the degree of development of the seminiferous epithelium. Cells undergoing degeneration were a common finding through the initial stages of spermatogenesis, coincident to the presence of sperm abnormalities and foreign cells in semen. By day 180 and onwards, both histology and seminal picture normalized. It is concluded that, under these rearing conditions, the onset of puberty (expressed as morphologically established spermatogenesis) in Corriedale ram lambs is attained at 180-216 days of age when they reach 23 cm of scrotal circumference and 191 g of testis weight. The finding of a high correlation between these parameters (r: 0.93) confirms the usefulness of the measurement of scrotal circumference during clinical examination of ram lambs in this breed.
Sujet(s)
Élevage , Ovis/physiologie , Spermatogenèse/physiologie , Analyse de variance , Aliment pour animaux , Phénomènes physiologiques nutritionnels chez l'animal , Animaux , Climat , Mâle , Poaceae , Dosage radioimmunologique/médecine vétérinaire , Saisons , Testostérone/sang , Facteurs temps , UruguayRÉSUMÉ
The ultrastructure of the tight junctional complex of pubertal ram lamb Sertoli cells was studied in immersion-fixed samples and related to clinical data and a light microscopical classification of the degree of spermatogenesis attained in the corresponding seminiferous tubule. Although the process followed the general mammalian developmental trend for tight junction complex formation, 2 unusual ultrastructural features were detected: the presence of an active Golgi complex during the early stage of tight junction formation and the transitory presence of ribosomes on both faces of the ectoplasmic cisternae bordering the developing junctions. The significance of these findings is discussed.