Identification, Development, and Regional Distribution of Thymidylate Synthetase in Adult Rabbit Brain

Abstract: The development and regional distribution of thymidylate synthetase (TS) (EC 2.1.1.45) in rabbit brain were determined. After optimization of the assay for brain, TS activity in brain was measured by a nonspecific (³H₂O release) and specific method. The specific method involved the conversion of [6-³H]deoxyuridine monophosphate (dUMP) to [³H]thymidine phosphate and the subsequent identification of [³H]thymidine. The specific activity of the enzyme in whole brain of newborn rabbits declined from 10.35 ± 1.17 units/mg protein to 0.71 ± 0.09 units/mg protein at 10–12 weeks of age. Two-year-old rabbits had 0.81 ± 0.04 units/mg protein. The decline in specific activity with age was not due to an inhibitor of TS activity or a change in the K_m for dUMP. The K_m for dUMP of the unpurified enzyme in the brains of both 10-day-old and young adult rabbits was 0.8 μ m . In young adult rabbits (3 months) the specific activity of TS was similar in the various regions of the brain tested except for the cerebellum, which had 40% higher specific activity than the whole brain. The results show that TS is widely distributed in adult rabbit brain, and, although the activity declines with age, it stabilizes at adult levels at 3 months of age.     

Immunodot detection of nisin Z in milk and whey using enhanced chemiluminescence

A highly specific antisera was produced in New Zealand white rabbits against nisin Z, a 3400 Da bacteriocin produced by Lactococcus lactis ssp. lactis biovar. diacetylactis UL 719. A dot immunoblot assay was then developed to detect nisin Z in milk and whey. As few as 1·5 10⁻¹ international units per ml (IU ml⁻¹), corresponding to 0·003 μg ml⁻¹ of pure nisin Z, were detected in carbonate-bicarbonate buffer within 6 h using chemiluminescence. When milk and whey samples were tested, approximately 0·155 μg ml⁻¹ (7·9 IU ml⁻¹) of nisin Z was detected. The detection limit obtained was lower than that of traditional methods including microtitration and agar diffusion.     

Experimental Infection of Nosema sp. Ghose in an Unnatural Host Lesioderma sericorne

ABSTRACT. Various doses of a microsporan parasite, Nosema sp., were fed to third and fourth instar larvae of Lesioderma sericorne that infested different types of stored grains. A spore dose of 3 × 10³ spores/individual resulted in a 39% infection rate, reduction in larval and adult weights, and mean spore concentrations of 1.28 ± 0.2 × 10⁸ spores/larva and 1.1 ± 0.2 × 10⁸ spores/adult. At the above dose, mortality was not well marked (about 35% in larvae and 25% in adults). At 3 × 10⁴ spores/individual, the rate of mortality increases to 80% in larvae and 60% in adults. However, more of the pest population (88% of larvae and 73% of adults) died at a dose of 3 × 10⁵ spores/individual. This dose produced mean spore concentrations of 3.91 ± 0.2 × 10⁸ spores/larva and 2.89 ± 0.2 × 10⁸ spores/adult. Insect death was caused by heavy damage to gut epithelia and fat bodies.     

Some mechanical properties of the flagellar-pedicellar suspension in Aedes aegypti (L.)

Abstract The rotation of the flagellum of male and female Aedes aegypti (L.) was measured in a wind tunnel at wind speeds between 0 and 116cms^-1. Although the surface area of the female flagellum is only 10% of the male, it rotates through double the angle of the male flagellum for the same increase in wind speed. The larger rotation of the female flagellum could result from: a reduced resistive torque in the flagellar-pedicellar suspension; a larger antennal-positioning reflex; a smaller initial angle of attack. The resistive torque in the flagellar-pedicellar suspension was measured in both male and female Aedes aegypti. In ten adult females the average torque at 8^o rotation was 2.70 ± 1.33 times 10^-10Nm and in ten males it was 6.46 ± 3.46 times 10^-10Nm. The significance of these results is discussed with reference to the sexual dimorphism in the sensory tasks required of Johnston's organ in the two sexes.     

Morphological and functional alterations induced in trout intestine by dietary cadmium and lead

Effects of oral administration of lead and cadmium on structure and function of trout intestine were investigated in Salmo gairdneri. Fish were fed either cadmium (5 mg kg*¹ fish per day) or lead (10 mg kg⁻¹ fish p er day) and sacrificed after a 15 or 30 day treatment.
Levels of cadmium and lead in kidney, liver and spleen were significantly increased by the 15th day of treatment.
Following both cadmium and lead treatment, morphological observations showed an increased mucous cell activity, a disruption of intestinal brush border and an increased renewal rate of absorptive cells.
Influx (J_ms), outflux (J_sm) of chlorine and sodium ion and net fluxes were measured on perfused intestinal segments and ouabain-sensitive sodium, potassium-ATPase (Na, K-ATPase) activity was determined on intestinal scrapings. Cadmium did not alter either sodium chlorine transepithelial fluxes or sodium, potassium-ATPase activity, but lead did. In the middle intestine, lead modified significantly transepithelial sodium and chlorine fluxes (J_ms Na: 3.21 ± 0.34 to 1.79 ± 0.29 and J_ms Cl: 3.32 ± 0.34 to 1.86 ± 0.22μmol h⁻¹ cm⁻², n = 6) after 30-day diet. J_net remain unchanged and Na, K-ATPase activity decreased. In the posterior intestine, lead altered only J_ms Na (1.95 ± 0.31 to 0.37 ± 0.09μmol h⁻¹ cm⁻²) and J_ms Cl. Consequently J_nes were also decreased.
So, although both cadmium and lead induce morphological disorders in the middle and in the posterior intestine of the trout, they have different effects on the absorption mechanisms of ions.     

Pyruvate Carboxylase Activity in Primary Cultures of Astrocytes and Neurons

Abstract: The activity of the pyruvate carboxylase was determined in brains of newborn and adult mice as well as primary cultures of astrocytes, of cerebral cortex neurons, and of cerebellar granule cells. The activity was found to be 0.25 ± 0.14, 1.24 ± 0.07, and 1.75 ± 0.13 nmol · min⁻¹· mg⁻¹ protein in, respectively, neonatal brain, adult brain, and astrocytes. Neither of the two types of neurons showed any detectable enzyme activity (i.e., < 0.05 nmol · min⁻¹· mg⁻¹). It is therefore concluded that pyruvate carboxylase is an astrocytic enzyme.     

Direct and indirect evidence for polypeptide absorption by the teleost gastrointestinal tract

The ability of the gastrointestinal tract of chinook salmon, Oncorhynchus tshawytscha , to absorb polypeptides in vivo was investigated by reference to the appearance of orally administered adrenocorticotropic hormone (ACTH) within the blood of fish previously treated with dexamethasone (3μg g^−l body weight) in order to suppress endogenous ACTH secretion. Further, since cortisol presence within plasma is dependent upon the availability of ACTH, dexamethasone blockade of endogenous ACTH secretion, in conjunction with subsequent measurements of plasma cortisol levels, provides a means by which biological patency of absorbed exogenous ACTH may be demonstrated. Levels of ACTH and cortisol in plasma of dexamethasone-treated salmon were therefore measured for a period of 360 min immediately following oral intubation of ACTH (15μg g⁻¹ body weight). Peak plasma presence of ACTH-like immunoreactivity (676.6 ± 33.6 pg ml⁻¹ plasma) and cortisol (227.1 ± 29.0 ng ml⁻¹ plasma) were recorded 120 min after ACTH administration. Results from the experimental groups were compared to those of 15 control treatments. Since administration of ACTH to chinook salmon elicited a consistent and significant elevation in not only plasma ACTH but also cortisol presence, it is contended that the salmonid gut expresses an ability to absorb polypeptides of dietary origin. The significance of these findings with respect to the oral administration of biologically active peptides and proteins to fish of importance to aquaculture is discussed.     

Seasonal patterns in water, sodium and energy turnover in free-living echidnas, Tachyglossus aculeatus (Mammalia: Monotremata)

The energetics of an egg-laying mammal, the echidna ( Tachyglossus aculeatus ), were studied in the wild by means of isotope turnover techniques. Water and sodium influx rates were highest in summer (47.7±15.3 ml kg^-1 day^-1 and 1.20±0.52 mmol kg^-1 day^-1, respectively) and associated with high metabolic rates (0.509±0.048 ml CO₂ g^-1 h^-1). Water and sodium influxes and metabolic rates were lowest in May and June (7.8±6.4 ml H₂O kg^-1 day^-1, 0.21±0.12 mmol Na kg^-1 day^-1 and 0.205±0.129 ml CO₂ g^-1 h^-1, respectively). These low rates in late autumn/early winter are associated with reduced activity, the animals spending substantial periods of time in torpor. The comparatively low isotope turnover rates of echidnas are a consequence of their diet; ants and termites which have low mass-specific energy contents.     

Heterogeneous Na+ Sensitivity of Na+,K+-ATPase Isoenzymes in Whole Brain Membranes

Abstract: The Na⁺ sensitivity of whole brain membrane Na⁺,K⁺-ATPase isoenzymes was studied using the differential inhibitory effect of ouabain (α1, low affinity for ouabain; α2, high affinity; and α3, very high affinity). At 100 m M Na⁺, we found that the proportion of isoforms with low, high, and very high ouabain affinity was 21, 38, and 41%, respectively. Using two ouabain concentrations (10⁻⁵ and 10⁻⁷ M ), we were able to discriminate Na⁺ sensitivity of Na⁺, K⁺-ATPase isoenzymes using nonlinear regression. The ouabain low-affinity isoform, α1, exhibited high Na⁺ sensitivity [ K _a of 3.88 ± 0.25 m M Na⁺ and a Hill coefficient ( n ) of 1.98 ± 0.13]; the ouabain high-affinity isoform, α2, had two Na⁺ sensitivities, a high ( K _a of 4.98 ± 0.2 m M Na⁺ and n of 1.34 ± 0.10) and a low ( K _a of 28 ± 0.5 m M Na⁺ and an n of 1.92 ± 0.18) Na⁺ sensitivity activated above a thresh old (22 ± 0.3 m M Na⁺); and the ouabain very-high-affinity isoform, α3, was resolved by two processes and appears to have two Na⁺ sensitivities (apparent K _a values of 3.5 and 20 m M Na⁺). We show that Na⁺ dependence in the absence of ouabain is the result of at least of five Na⁺ reactivities. This molecular functional characteristic of isoenzymes in membranes could explain the diversity of physiological roles attributed to isoenzymes.     

Thermoregulation and metabolism in the Cape golden mole (Insectivora: Chrysochloris asiatica)

The Cape golden mole, Chrysochloris asiatica is an insectivore which excavates superficial foraging burrows as it searches for its food. It has a mean (±S.D.) resting metabolic rate (RMR) when newly captured of 1–17±0.17 cm³ O₂g^-1 h^-1 ( n = 14), within the thermoneutral zone (TNZ) of 30–32°C.
The body temperature (Tb) of the mole in the TNZ is low 32.9 ± 0.36 ( n = 14) and remains stable at ambient temperatures (Tas) from 28–32°C. Above 32°C (range 34–37°C), Tb increases albeit slightly to 36 ± 1.75°C ( n = 14). The conductance is high 0.27 ± 006cm³ O₂g^-1 h^-l°C^-1 ( n = 46) at the lower limit of thermoneutrality. The mean RMR at 9°C (the lowest Ta tested) was 4.82±11 cm³ O₂g^-1h^-1, which is 4.1 times that of the RMR in the TNZ.
At an ambient temperature of 9°C, three of the golden moles entered a state of torpor where the RMR was reduced from 5.9±0.56 to 10 1.0 ± 0.69cm³O₂g^-1h^-1.     

Reduction of caecal Listeria monocytogenes in Leghorn chicks following treatment with a competitive exclusion culture (PREEMPT™)

Day-of-hatch Leghorn chicks were treated by oral gavage with PREEMPT^™, a continuous-flow competitive exclusion culture containing broiler caecal bacteria, followed by an oral challenge with Listeria monocytogenes , to determine the effects of PREEMPT^™ on L. monocytogenes caecal colonization. Increased ( P < 0·001) concentrations of caecal propionic acid in control chicks compared with PREEMPT^™-treated chicks at 3 days of age were indicative of the establishment of the PREEMPT^™ bacteria. Caeca from control chicks at 7 days after the oral challenge with L. monocytogenes contained mean 3·4 ± 1·4 log cfu g⁻¹ of caecal content, while caeca from PREEMPT^™-treated chicks contained no detectable Listeria. Enrichment for L. monocytogenes resulted in 100 % of caeca from control chicks testing culture-positive for L. monocytogenes , while none of the caeca from PREEMPT^™-treated chicks were culture-positive. The results indicated that prophylactic treatment of newly hatched chicks with PREEMPT^™ significantly reduced caecal colonization by L. monocytogenes.     

Reaction of Muscimol with 4-Aminobutyrate Aminotransferase

Abstract: The reaction of muscimol as amino donor substrate for GABA transaminase (GABA-T) has been studied using enzyme purified from rabbit brain. Enzyme activity was assayed by measuring the glutamate produced using glutamate dehydrogenase. Kinetic parameters determined at 37°C were for GABA, K _m (app) = 1.92 ± 0.24 m M , specific activity = 7.33 ± 0.27 μmol/min/mg ( k _cat= 13.7s⁻¹), and for muscimol, K _m (app) = 1.27 ± 0.15 m M , specific activity = 0.101 ± 0.009 μmol/min/mg ( k _cat= 0.19s⁻¹). Addition of muscimol to the enzyme caused the spectral changes associated with conversion of the pyridoxaldimine form to the pyridoxamine form, and the first-order rate constant for the reaction showed a dependence on muscimol concentration that followed saturation kinetics, with a K = 1.1 ±0.18 m M and k _max= 0.065 ± 0.004 s⁻¹ (19°C). The rate of spectral change observed on addition of muscimol to ornithine transaminase was extremely slow—at least an order of magnitude slower than that seen with GABA-T.     

Growth, diet and metabolism of common wolf–fish in the North Sea, a fast–growing population

The von Bertalanffy growth parameters for common wolf–fish Anarhichas lupus in the North Sea were: male: L ∞=111·2 cm, t ₀=–0·43 and K =0·12; and female: L ∞=115·1 cm, t ₀=–0·39 and K =0·11, making this the fastest growing stock reported. Resting metabolic rates (RMR±S.E.) and maximum metabolic rates (MMR±S.E.) for six adult common wolf–fish (mean weight, 1·39 kg) at 5° C were 12·18±1·6 mg O₂ kg^–1 h^–1 and 70·65±7·63 mg O₂ kg^–1 h^–1 respectively, and at 10° C were 25·43±1·31 mg O₂ kg^–1 h^–1 and 113·84±16·26 mg O₂ kg^–1 h^–1. Absolute metabolic scope was 53% greater at 10° C than at 5° C. The diet was dominated by Decapoda (39% overall by relative occurrence), Bivalvia (20%) and Gastropoda (12%). Sea urchins, typically of low energy value, occupied only 7% of the diet. The fast growth probably resulted from summer temperatures approximating to the optimum for food processing and growth, but may have been influenced by diet, and reduced competition following high fishing intensity.     

Relationships between biovolume and carbon and nitrogen content of bacterioplankton

Abstract Cell volume, carbon and nitrogen content were determined for bacteria grown in batch cultures in water samples collected at five localities in western Florida, USA. Cultures were set up by inoculating 0.2 μm filtered water with 2.5 to 7.0% of 1.0 μm filtered water. Biovolumes of the bacteria were measured by epifluorescence photomicrography. Bacterial carbon and nitrogen contents were determined with a CHN analyser. During incubations, bacterial volumes doubled from 0.070±0.037 μ m³(mean ± S.E.) to 0.153 ± 0.036 μ m³ at early stationary phase. Bacterial C:N ratios ranged between 2.8 and 10.3, with a mean of 6.5, and were inversely correlated with cell volumes. Conversion factors for volume to carbon and nitrogen content were relatively high and variable, ranging from 0.21 to 161 pg C μm⁻³ (mean: 0.72 pg C μm⁻³) and from 0.05 to 0.25 pg N μm⁻³ (mean: 0.12 pg N μm⁻³). Small cells contained more C and N per unit volume than did large cells. The data suggested that biovolume to biomass conversion factors may be higher than previously thought and may be highly variable both temporally and geographically.     

Surface free energies of oral streptococci

Abstract The surface free energies ( γ _b) of a variety of oral streptococci were determined from contact angle measurements on bacterial deposits, using the concept of dispersion and polar components. At least four strains of each species were tested. Strains of Streptococcus mutans, S. sanguis and S. salivarius possessed relatively high surface free energies (103 ± 12 mJ · m⁻²) and at the species level no significant difference was found. In contrast, the strains of S. mitis had remarkably low surface free energies (45 ± 14 mJ · m⁻²). S. milleri appeared to be a heterogeneous species, showing surface free energies over a range of 32–119 mJ · m⁻². No significant differences were observed between laboratory strains and strains freshly isolated from the oral cavity.     

Water-relation parameters of giant and normal cells of Capsicum annuum pericarp

Abstract. Measurements of the water-relation parameters of the giant subepidermal cells (volume, V = 0.119 to 1.658 mm³; = 0.53±0.35 mm³, SD, n = 23) and the smaller mesocarp parenchyma cells ( V = 0.10 to 0.79×10⁻³ mm³; = 0.36±0.27×10⁻³ mm³, SD, n = 6) of the inner pericarp surface of Capsicum annuum L. were made using the Jülich pressure probe. The volumetric elastic modulus ɛ for the large cells was between 1.5 and 27 MPa for a pressure range of 0.09 to 0.41 MPa. For the small cells ɛ was 0.1 to 0.6 MPa for a pressure range of 0.22 to 0.39 MPa. The turgor pressure P , the half-time of water exchange T _1/2, and the hydraulic conductivity L _p were as follows, with SD and number of replicates: large cells, P = 0.27±0.06 MPa (23), T _1/2=2.7±2.2 s (46), L _p=5.8±3.7 pm s⁻¹ Pa⁻ (46); small cells, P = 0.33±0.07 MPa (6), T _1/2= 33±10s (12), L _p=0.21±0.07 pm s⁻¹ Pa⁻¹ (12). The determination of these basic water-relation parameters is considered as a prerequisite for future ecotoxicological and phytopathological studies. The differences between the large and the small cells are discussed in relation to a desirable biophysical definition of succulence. Further, for the large cells a pressure and volume dependence of ɛ was demonstrated.     

Oxygen consumption of East Siberian cod: no support for the metabolic cold adaptation theory

Standard metabolic rate ( R _s) at 2°C of eight East Siberian cod Arctogadus borisovi , caught in West Greenland, body mass of 601.5 ± 147.6 g (mean ± s.D.), was 40.9 ± 5.9 mg O₂ kg^-1 h^-1 and 59.0 ± 6.6mg O₂ kg^-1 h^-1 when extrapolated to a standardized 100 g fish. R _s was compared with three other Gadidae, to test the theory of metabolic cold adaptation (MCA). There was no evidence of MCA in the family.     

Seasonal differences in routine oxygen consumption rates of the bonnethead shark

Routine oxygen consumption rates of bonnethead sharks, Sphyrna tiburo , increased from 141·3±29·7 mg O₂ kg⁻¹ h⁻¹ during autumn to 218·6±64·2 mg O₂ kg⁻¹ h⁻¹ during spring, and 329·7±38·3 mg O₂ kg⁻¹ h⁻¹ during summer. The rate of routine oxygen consumption increased over the entire seasonal temperature range (20–30° C) at a Q ₁₀=2·34.     

Membrane proteins with molecular masses of 88, 90 and 150 kDa are responsible for binding of human immunoglobulin G Fc fragment to the native cells of Mycoplasma salivarium

Abstract Growth of Pyrococcus furiosus was studied in batch cultures with cellobiose, maltose and pyruvate as limiting substrates. Fermentation mass balances of all conversions were complete. These data suggested that the pathway for conversion of the disaccharides is essentially the same. The molar growth yields on maltose and cellobiose were about equal (±50 g cell dry weight (cdw) per mol disaccharide). The molar growth yield on pyruvate was ±6.3 g cdw mol⁻¹. Growth yields were influenced by the hydrogen partial pressure. When P. furiosus was co-cultured with a methanogen a yield of 56 g cdw mol⁻¹ disaccharide and 8.6 g cdw mol⁻¹ pyruvate was obtained. When the data were interpreted according to the proposed pyroglycolytic pathway, the calculated Y_ATP values were different for the various pH₂ conditions, suggesting that an additional energy conserving site may be present in the pathway.     

Annual changes of bacterial mortality due to viruses and protists in an oligotrophic coastal environment (NW Mediterranean)

The impact of viruses and protists on bacterioplankton mortality was examined monthly during 2 years (May 2005–April 2007) in an oligotrophic coastal environment (NW Mediterranean Sea). We expected that in such type of system, (i) bacterial losses would be caused mainly by protists, and (ii) lysogeny would be an important type of virus–host interaction. During the study period, viruses and grazers together were responsible for 50.6 ± 40.1% day⁻¹ of bacterial standing stock losses (BSS) and 59.7 ± 44.0% day⁻¹ of bacterial production losses (BP). Over the first year (May 2005–April 2006), protists were the principal cause of bacterial mortality, removing 29.9 ± 20.4% day⁻¹ of BSS and 33.9 ± 24.3% day⁻¹ of BP, whereas viral lysis removed 13.5 ± 17.0% day⁻¹ of BSS and 12.3 ± 12.3% day⁻¹ of BP. During the second year (May 2006–April 2007), viruses caused comparable bacterial losses (29.2 ± 14.8% day⁻¹ of BSS and 40.9 ± 20.7% day⁻¹ of BP) to protists (28.6 ± 25.5% day⁻¹ of BSS and 32.4 ± 20.0% day⁻¹ of BP). In 37% of cases higher losses of BP due to viruses than due to protists were found. Lysogenic infection was detected in 11 of 24 samplings. Contrary to our expectations, lytic infections dominated over the two years, and viruses resulted to be a significant source of bacterial mortality in this oligotrophic site.