Biosynthesis and release of juvenile hormone during the reproductive cycle of the ring-legged earwig

Corpora allata of adult female Euborellia annulipes, incubated in medium containing ³H-methionine, synthesized and released juvenile hormone III. Labelled material co-migrating with methyl farnesoate was also found, suggesting this as an intermediate in the pathway of juvenile hormone III production. Juvenile hormone was not appreciably stored in the glands, but was released into the medium. In normal medium, 93.6 ± 1.6% of the total juvenile hormone III synthesized was released and 96.5% ± 0.3 in medium supplemented with 60 μM farnesoic acid. The rate of juvenile hormone III biosynthesis/release in vitro remained constant for at least 8 hr for glands of different activities. The rate of juvenile hormone production was closely correlated with the gonadotrophic cycle. In females with previtellogenic ovarian follicles (0.26 ± 0.004 mm), hormone production was only 0.59 ± 0.13 fmol hr⁻/corpus allatum; production increased to 1.52 ± 0.25 fmol hr⁻¹/corpus allatum when basal follicles were growing rapidly, and remained high during the period of oviposition. By 3 days following oviposition when females were brooding clutches, hormone production had declined to 0.46 ± 0.13 fmol hr⁻¹/corpus allatum. The addition of 60 μM farnesoic acid to the medium enhanced juvenile hormone biosynthesis at each stage examined. Lastly, elevating the level of l-methionine in the medium also enhanced hormone biosynthesis. Maximal hormone production was 32.8 ± 10.9 fmol hr⁻¹/corpus allatum, at an l-methionine concentration of 51 μM.     

BUOYANCY REGULATION IN THE COLONIAL DIAZOTROPHIC CYANOBACTERIUM TRICHODESMIUM TENUE: ULTRASTRUCTURE AND STORAGE OF CARBOHYDRATE,POLYPHOSPHATE, AND NITROGEN1

Trichodesmium tenue Wille (1904) was examined using transmission electron microscopy to determine the role of carbohydrate, phosphorus, and nitrogen storage in buoyancy regulation. Carbohydrate storage area (mean = 2.06 ± 0.61 [SE] μm²; 6.62% of total cell area) in negatively buoyant colonies (NBCs) was significantly higher (P < 0.001) than in positively buoyant colonies (PBCs) (mean = 0.38 ± 0.06 μm²; 0.73%). Distinct diel periodicity of carbohydrate content was found in NBCs demonstrated by an increase from darkness to afternoon. Polyphosphate content was significantly higher (P < 0.001) in NBCs, with a mean of 0.44± 0.10 μm² (1.54%), as compared to PBCs, with a mean of 0.14 ± 0.05 μm² (0.24%). Polyphosphate content increased in NBCs from morning to evening, and PBCs had a 10% decrease from morning to afternoon. Calculations indicated that averaged effects of polyphosphate on increased cell density is approximately 20% of that from carbohydrate accumulation. Density contribution due to ballast weight of carbohydrate and polyphosphate indicated that NBCs were 12 times more dense than PBCs. Mean area of cyanophycin granules (N storage) was not significantly different between PBCs and NBCs. In conclusion, Trichodesmium tenue can regulate buoyancy by carbohydrate ballasting similar to that noted in limnetic cyanobacteria. Polyphosphate storage and possibly nitrogen storage products play a significant role in buoyancy regulation.     

Photosynthetic parameters of Ulmus minor plantlets affected by irradiance during acclimatization

In order to set up large-scale acclimatization protocols of micropropagated plants, an in-depth knowledge of their physiological responses during in vitro to ex vitro transfer is required. This work describes the photosynthetic performance of Ulmus minor micropropagated plants during acclimatization at high irradiance (HI; 200 ± 20 μmol m^?2 s^?1 or low irradiance (LI; 100 ± 20 μmol m^?2 s^?1). During this experiment, leaf pigment content, chlorophyll a fluorescence, gas exchange, stomata morphology, the activity of the Calvin cycle enzymes and saccharides were measured in persistent and new leaves. The results indicated that HI induces a higher photosynthetic performance compared to LI. Therefore, plants acclimatized under HI are likely to survive better after field transfer.     

The production and secretion of cortisol by baboon neonates.

The metabolic clearance rate (MCR), transfer constants (p), production (PR) and secretion (SR) rates of cortisol (F) andrortisone (E) were determined by the continuous infusion of {1,2³H}F and {4-¹⁴C}E into 5 neonates delivered prior to the parturition by cesarean section (164–179 days; term = 184 days) and into 5 newborns delivered spontaneously per vagina at term (166 – 187 days). In spontaneously delivered animals, MCR-E (X ± SE, 34.3 ± 7.0 1/day/kg was greater (P < 0.001) than MCR-F (14.9 ± 1.5 1/day/kg), pF to E (59.7 ± 8.9%) exceeded (P < 0.001) pE to F (17.8 ± 3.0%) and the percentage of F bound to serum proteins other than albumin (57.5 ± 6.2) was greater (P < 0.001) than that of E (27.0 ± 10.3) Although the serum E level (25.6 ± 3.6 μg/100 ml) was similar to that of F (33.5 ± 8.0 μg/100 ml), the PR-E (6.4 ± 1.3 μ/min/kg) was greater (P < 0.001) than PR-F (3.3 ± 0.5 μ/min/kg). Approximately eighty-five percent of the E and 65% of the F produced orginated by secretion.In animals delivered by cesarean section, the serum F concentration (32.4 ± 6.7 μ/100ml), pE to F (13.4 ± 2.8%) pF to E (80.0 ± 12.2%) PR-E (4.5 ± 0.2 μ/min/kg) and SR-E (3.9 ± 0.3 μ/min/kg) were not different from values for spontaneously delivered animals. Serum E levels (35.9 ± 1.6 μ/100 ml) were higher but MCR-F (6.7 ± 0.6 1/day/kg) and MCR-E (18.2 ± 0.41/day/kg) lower in neonates delivered by cesarean section. Serum Cortisol binding capacity (μg F bound/100 ml) was greater (P < 0.025) in neonates delivered by cesarean section (23.6 ± 2.6) than in spontaneously delivered animals (14.4 ± 2.0). As a result of these changes in F and E dynamics, PR-F (1.4 ± 0.3 μ/min/kg) and SR-F (0.9 ± 0.2 μ/min/kg) in neonates delivered by cesarean section were lower (P< 0.01) than corresponding values in spontaneously delivered newborns.It is concluded that the greater F secretion in animals delivered spontaneously than those delivered by cesarean section probably results from increased fetal adrenal 3β-hydroxysteroid dehydrogenase-isomerase activity, which as previously reported, occurs in late gestation in this species.     

Morphometric characterization of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa using computer-assisted spermatozoa analysis (ASMA)

As part of a larger study on sperm quality and cryopreservation methods, the present study characterized the head morphometry of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa, using both scanning electron microscopy (SEM) and computer‐assisted morphology analysis (ASMA). The latter method has been used rarely in fish and this is its first application on sharpsnout sea bream and gilthead sea bream spermatozoa. Results obtained using SEM are expensive and time‐consuming, while ASMA provides a faster and automated evaluation of morphometric parameters of spermatozoa head. For sharpsnout sea bream spermatozoa, similar head measurement values were obtained using both ASMA and SEM, having a mean ± standard error length of 2.57 ± 0.01 μm vs 2.54 ± 0.02 μm, width of 2.22 ± 0.02 μm vs 2.26 ± 0.04 μm, surface area of 4.44 ± 0.02 μm² vs 4.50 ± 0.04 μm² and perimeter of 7.70 ± 0.02 μm vs 7.73 ± 0.04 μm using ASMA and SEM, respectively. Although gilthead sea bream spermatozoa were found to be smaller than those of sharpsnout sea bream, spermatozoal head morphometry parameters were also found to be similar regardless of evaluation method, having a mean head length of 1.97 ± 0.01 μm vs 1.94 ± 0.02 μm, head width of 1.80 ± 0.01 μm vs 1.78 ± 0.02 μm, surface area of 3.16 ± 0.03 μm² vs 3.18 ± 0.06 μm² and perimeter of 6.52 ± 0.04 μm vs 6.56 ± 0.08 μm using ASMA and SEM, respectively. The results demonstrate that ASMA can be considered as a reliable technique for spermatozoal morphology analysis, and can be a useful tool for studies on fish spermatozoa, providing quick and objective results.     

Giemsa-based cytological assessment of area,shape and nucleus:cytoplasm ratio of goblet cells of rabbit bulbar conjunctiva

Goblet cells were visualized in impression cytology specimens from bulbar conjunctiva of the rabbit eye using Giemsa staining. Highly magnified images were used to generate outlines of the goblet cells and their characteristic eccentric nuclei. Using sets of 10 cells from 15 cytology specimens, I found that the longest dimension of the goblet cells averaged 16.7 ± 2.3 μm, the shortest dimension averaged 14.4 ± 1.8 μm and the nucleus averaged 6.3 ± 0.8 μm. The goblet cells were ellipsoid in shape and the longest:shortest cell dimension ratio averaged 1.169 ± 0.091. The goblet cell areas ranged from 108 to 338 μm² (average 193 ± 50 μm²). The area could be predicted reliably from the longest and shortest dimensions (r² = 0.903). The areas of goblet cell nuclei were 15–58 μm² (average 33 ± μm²) and the nucleus:cytoplasm area fraction was predictably greater in smaller goblet cells and less in the larger goblet cells (Spearman correlation = 0.817). The nuclei were estimated to occupy an average of 9.5% of the cell volume. The differences in size, shape and nucleus:cytoplasm ratio may reflect differences in goblet cell maturation.     

Size-fractionated chlorophyll a and primary productivity in the Chukchi Sea and its northern Chukchi Plateau

Investigations on chlorophyll a and primary productivity were carried out in the Chukchi Sea and its northern Chukchi Plateau during the 2^nd Chinese National Arctic Research Expedition in the summer of 2003. The results showed that chlorophyll a concentrations were 0.009–30.390 μg/dm³ at the surveyed waters; the surface chlorophyll a concentrations were 0.050–4.644 μg/dm³ and the average value was (0.875±0.981) μg/dm³ in the surveyed area. In the Chukchi Sea Shelf, chlorophyll a concentrations at the depth from 10 m to bottom were higher than that in the surface water, and the concentrations were lower at the depth below 75 m in the Chukchi Plateau. Chlorophyll a concentrations descended in 3 sequential samplings on Transect R, with average values of (2.564±1.496) μg/dm³, (1.329±0.882) μg/dm³ and (0.965±0.623) μg/dm³, respectively. The potential primary productivity ((2.305± 1.493) mgC/(m³·h)) in the Chukchi Sea was higher than that ((0.527±0.374) mgC/(m³·h)) in the Chukchi Plateau. The results of the size-fractionated chlorophyll a and primary productivity showed that microplankton accounted for the majority of the total chlorophyll a (63.13%) and primary productivity (65.16%) at the survey stations. The contributions of the nanoplankton and picoplankton to the total chlorophyll a and primary productivity were roughly the same.     

Contribution of terrestrial invertebrates to the annual resource budget for salmonids in forest and grassland reaches of a headwater stream

1. The annual input, contribution to the diet of salmonids, and quantitative input of terrestrial invertebrates to four reaches with contrasting forest (n=2) and grassland riparian vegetation (n=2) were compared in a Japanese headwater stream.
2. The annual input of terrestrial invertebrates falling into the forest reaches (mean±1 SE=8.7×10³±0.3×10³ mg m^?2 year^?1) was 1.7 times greater than that in the grassland reaches (5.1×10³±0.8×10³ mg m^?2 year^?1), with clear seasonality in the daily input of invertebrates in both vegetation types. The daily input, however, differed between the vegetation types only in summer, when it rose to a maximum in both vegetation types.
3. Fish biomass also differed among the seasons in both vegetation types, being less in the grassland reaches. The contribution of terrestrial invertebrates to the salmonid diet in the forest and grassland reaches was 11 and 7% in spring, 68 and 77% in summer, 48 and 33% in autumn, and 1 and 1% in winter, respectively. The prey consumption rate of fish, which was similar between the vegetation types, increased with stream temperature and was highest in summer. Terrestrial invertebrates supported 49% (mean±1 SE=5.3×10³±0.4×10³ mg m^?2 year^?1) of the annual, total prey consumption (10.9×10³±1.7×10³ mg m^?2 year^?1) by salmonids in the forest and 53% (2.0×10³±0.3×10³ mg m^?2 year^?1) (3.8×10³±0.6×10³ mg m^?2 year^?1) in the grassland reaches.
4. Salmonids were estimated to consume 51 and 35% of the annual total (falling plus drift) input of terrestrial invertebrates in the forest and grassland reaches, respectively. The input of terrestrial invertebrates by drift, however, was almost equal to the output in both vegetation types, suggesting that the reach‐based, in‐stream retention of terrestrial invertebrates almost balanced these falling in.
5. Difference in the riparian vegetation, which caused spatial heterogeneity in the input of terrestrial invertebrates, could play an important role in determining the local distribution of salmonids.     

Ultrafiltration-light absorption spectrophotometric determination of silicon in blood

An ultrafiltration-light absorption spectrometric method for soluble molybdate-reactive silicon was assessed and applied to bovine and ovine blood plasma and sera, giving precise analytical results. Interfering protein above molecular weight 10,000–25,000 was removed by ultrafiltration, and silicon in ultrafiltrates was quantitated by measuring light absorption at 810 nm of the 1,2,4-aminonaphthol sulfonic acid/ascorbic acid-reduced silicomolybdate. Chemical interferences on the color-forming reaction of remaining blood components were tested by measuring recoveries of silicon added to real blood plasma samples and to synthetic blood plasma solutions, the latter containing typical levels of the major ions Na⁺, K⁺, Ca²⁺, HCO₃^?, and Cl^?, together with varying quantities of the potential interferants (amount per analytical reaction): phosphate (0–0.5 mg P), ferric ion (0–3 mg), fluoride (0–1.25 mg), vanadate (0–0.5 mg V), arsenate (0–10 μg As), and germanate (0–0.5 μg Ge). The mean recovery of added 0.8–9 μg silicon/g of bovine and ovine plasma was 97.7% (SE = 1.0, n = 17); the mean recovery of 1 and 5 μg silicon from synthetic blood plasma solutions with interferant levels up to 50-fold that in normal plasma was 99.2% (SE = 0.3, n = 47). Silicon concentrations found in bovine and ovine blood plasma and sera were typically around 7 μg/ml with procedural reagent blanks consistently low at a mean of 0.12 μg/test (SD = 0.011, n = 20). The silicon level in Center for Disease Control bovine serum (reference specimen Lot R-2274) was found to be (mean ± SE, n = 10) 1.147 ± 0.013 μg/g or 1.172 ± 0.013 μg/ml (25°C). The method detectivity (detection limit) was estimated at 0.03 μg.     

The effects of synthetic estrogens on the metabolic clearance and production rates of estrone and estradiol

The metabolic clearance rates (MCR) of estrone (1) and estradiol were determined by pulse injections and constant infusions of ³H-estrone and ³H-estradiol in seven women taking mestranol-containing compounds and in seven women taking ethinyl estradiol-containing compounds. These results were compared with the results previously obtained in our laboratory (2, 3, 4, 5) in comparable women not taking these compounds. In the women taking mestranol the mean (± SE) MCR for estradiol, 750 ± 600 1/day/m², was similar to our normal mean value, 790 ± 30 1/day/m². However, the mean MCR for estrone was less 1,010 ± 60 1/day/m² than that in normals 1,230 ± 30 1/day/m². In the women taking ethinyl estradiol the mean MCR for estradiol, 1,070 ± 60 1/day/m² was significantly (P < 0.01) greater than the normal value. The mean MCR for estrone, 1,180 ± 80 1/day/m² was not different from the normal.Using an immunoassay to measure the concentrations of estradlol and. estrone in plasma, the mean level of estradlol in mestranol users was 40 ± 7 Pg/ml and in ethinyl estradlol users 69 ± 4 pg/ml.The mean calculated production rate for estradlol in the mestranol users was 47 ± 8 μg/day and in the ethinyl estradlol users was 120 ± 22 μg/day. The mean calculated, production rates for estrone were 71 ± 12 and 93 ± 12 μg/day in the respective groups.Thus while mestranol appears to have little effect on endogenous estrogen metabolism, the use of ethinyl estradiol appears to increase the MCR of estradlol, but not of estrone. The MCR of estradlol returns to the normal range when ethinyl estradlol is stopped.     

Some electrophysiological and permeability properties of the mouse egg

Certain electrophysiological and ionic properties of the mouse egg (CF-1 and BDF 12–18 hr post ovulation) have been investigated. Membrane potential (?14 ± 0.4 mV, ± SE, inside negative), membrane resistance (2610 ± 38 ohm·cm²), and membrane capacitance (1.6 ± 0.03 μF cm^?2) have been determined by means of intracellular microelectrode recording techniques. Membrane potential and related parameters are stable for extended periods of time upon impalement and the magnitude of the cell membrane potential has been demonstrated to be sensitive to alteration in external sodium. The electrophysiological studies in conjunction with measurements of unidirectional potassium fluxes using isotope tracer-techniques have allowed determination of membrane permeability to potassium (8 × 10^?8 cm sec^?1) and membrane potassium conductance (25 μmho cm^?2). Furthermore, the use of tracer flux techniques has indicated that the exchangeable fraction of intracellular potassium is 204 ± 14 mM. This represents the bulk of egg potassium (222 ± 19 mM as determined from flame photometry). Studies of unidirectional potassium efflux have indicated that its movement out of the egg is made up of at least two components; an external potassium-independent potassium efflux and external potassium-dependent efflux, the latter possibly representing a potassium exchange mechanism. The combined electrophysiological and tracer-flux data indicate that only a small portion of the total membrane conductance is composed of potassium conductance at this stage of development. This and the fact that the membrane potential is far from the potassium equilibrium potential are similar to observations made on mature eggs of several other species.     

N2O, CH4 and CO2 emissions from seasonal tropical rainforests and a rubber plantation in Southwest China

The main focus of this study was to evaluate the effects of soil moisture and temperature on temporal variation of N₂O, CO₂ and CH₄ soil-atmosphere exchange at a primary seasonal tropical rainforest (PF) site in Southwest China and to compare these fluxes with fluxes from a secondary forest (SF) and a rubber plantation (RP) site. Agroforestry systems, such as rubber plantations, are increasingly replacing primary and secondary forest systems in tropical Southwest China and thus effect the N₂O emission in these regions on a landscape level. The mean N₂O emission at site PF was 6.0 ± 0.1 SE μg N m⁻² h⁻¹. Fluxes of N₂O increased from <5 μg N m⁻² h⁻¹ during dry season conditions to up to 24.5 μg N m⁻² h⁻¹ with re-wetting of the soil by the onset of first rainfall events. Comparable fluxes of N₂O were measured in the SF and RP sites, where mean N₂O emissions were 7.3 ± 0.7 SE μg N m⁻² h⁻¹ and 4.1 ± 0.5 SE μg N m⁻² h⁻¹, respectively. The dependency of N₂O fluxes on soil moisture levels was demonstrated in a watering experiment, however, artificial rainfall only influenced the timing of N₂O emission peaks, not the total amount of N₂O emitted. For all sites, significant positive correlations existed between N₂O emissions and both soil moisture and soil temperature. Mean CH₄ uptake rates were highest at the PF site (−29.5 ± 0.3 SE μg C m⁻² h⁻¹), slightly lower at the SF site (−25.6 ± 1.3 SE μg C m⁻² h⁻¹) and lowest for the RP site (−5.7 ± 0.5 SE μg C m⁻² h⁻¹). At all sites, CH₄ uptake rates were negatively correlated with soil moisture, which was also reflected in the lower uptake rates measured in the watering experiment. In contrast to N₂O emissions, CH₄ uptake did not significantly correlate with soil temperature at the SF and RP sites, and only weakly correlated at the PF site. Over the 2 month measurement period, CO₂ emissions at the PF site increased significantly from 50 mg C m⁻² h⁻¹ up to 100 mg C m⁻² h⁻¹ (mean value 68.8 ± 0.8 SE mg C m⁻² h⁻¹), whereas CO₂ emissions at the SF and RP site where quite stable and varied only slightly around mean values of 38.0 ± 1.8 SE mg C m⁻² h⁻¹ (SF) and 34.9 ± 1.1 SE mg C m⁻² h⁻¹ (RP). A dependency of soil CO₂ emissions on changes in soil water content could be demonstrated for all sites, thus, the watering experiment revealed significantly higher CO₂ emissions as compared to control chambers. Correlation of CO₂ emissions with soil temperature was significant at the PF site, but weak at the SF and not evident at the RP site. Even though we demonstrated that N and C trace gas fluxes significantly varied on subdaily and daily scales, weekly measurements would be sufficient if only the sink/ source strength of non-managed tropical forest sites needs to be identified.     

High heterotrophic CO<Subscript>2</Subscript> emissions from a Malaysian oil palm plantations during dry-season

Tropical peatlands are currently being rapidly cleared and drained for the establishment of oil palm plantations, which threatens their globally significant carbon sequestration capacity. Large-scale land conversion of tropical peatlands is important in the context of greenhouse gas emission factors and sustainable land management. At present, quantification of carbon dioxide losses from tropical peatlands is limited by our understanding of the relative contribution of heterotrophic and autotrophic respiration to net peat surface CO₂ emissions. In this study we separated heterotrophic and autotrophic components of peat CO₂ losses from two oil palm plantations (one established in ‘2000’ and the other in 1978, then replanted in ‘2006’) using chamber-based emissions sampling along a transect from the rooting to non-rooting zones on a peatland in Selangor, Peninsular Malaysia over the course of 3 months (June–August, 2014). Collar CO₂ measurements were compared with soil temperature and moisture at site and also accompanied by depth profiles assessing peat C and bulk density. The soil respiration decreased exponentially with distance from the palm trunks with the sharpest decline found for the plantation with the younger palms with overall fluxes of 1341 and 988 mg CO₂ m^?2 h^?1, respectively, at the 2000 and 2006 plantations, respectively. The mean heterotrophic flux was 909 ± SE 136 and 716 ± SE 201 mg m^?2 h^?1 at the 2000 and 2006 plantations, respectively. Autotrophic emissions adjacent to the palm trunks were 845 ± SE 135 and 1558 ± SE 341 mg m^?2 h^?1 at the 2000 and 2006 plantations, respectively. Heterotrophic CO₂ flux was positively related to peat soil moisture, but not temperature. Total peat C stocks were 60 kg m^?2 (down to 1 m depth) and did not vary among plantations of different ages but SOC concentrations declined significantly with depth at both plantations but the decline was sharper in the second generation 2006 plantation. The CO₂ flux values reported in this study suggest a potential for very high carbon (C) loss from drained tropical peats during the dry season. This is particularly concerning given that more intense dry periods related to climate change are predicted for SE Asia. Taken together, this study highlights the need for careful management of tropical peatlands, and the vulnerability of their carbon storage capability under conditions of drainage.     

Characterization of mannosyl transferases during the pupal instar of Stomoxys calcitrans (L)

Particulate fractions (10,000g) from pupae of Stomoxys calcitrans transfer [¹⁴C]-mannose from GDP-[¹⁴C]-mannose to dolichol monophosphate and proteins. Production of the mannosyl lipid was inhibited by Mn²⁺, UDP, GMP, GDP, and EDTA. The insect growth regulator diflubenzuron had no effect on mannosyl transferase activity. Dolichol monophosphate and Mg²⁺ stimulated mannosyl transferase activity. The mannosyl lipid product was identified as mannosyl-phosphoryl-dolichol (Man-P-Dol). The apparent K_m and V_max values for the formation of Man-P-Dol using GDP-[¹⁴C]-Man while holding dolichol phosphate constant were 2.4 ± 0.9 μM and 9.4 ± 2.3 pmol Man-P-Dol·min^?1·mg^?1 protein, respectively. The apparent K_m and V_max values using dólichol phosphate while holding GDP-Man constant were 2.2 ± 1.2 μM and 18.5 ± 1.7 pmol Man-P-Dol·min^?1·mg^?1 protein.     

CARRIER MEDIATED BLOOD-BRAIN BARRIER TRANSPORT OF CHOLINE AND CERTAIN CHOLINE ANALOGS

Blood-brain barrier (BBB) transport of choline and certain choline analogs was studied in adult and suckling rats, and additionally compared in the paleocortex and neocortex of adult rats. Saturable uptake was characterized by a single kinetic system in all cases examined, and in adult rat forebrains we determined a K_m= 442 ± 60 μM and V_max= 10.0 ± 0.6 nmol min^-1 g^-1. In 14–15-day-old suckling forebrains a similar K_m (= 404 ± 88 μM) but higher V_max (= 12.5 ± 1.5 nmol min^-1 g^-1) was determined. When choline uptake was compared in two regions of the forebrain, similar Michaelis-Menten constants were determined but a higher uptake velocity was found in the neocortex (i.e. neocortex K_m= 310 ± 103 μM and V_max= 12.6 ± 2.8 nmol min^-1g^-1; paleocortex K_m= 217 ± 76 μM and V_max= 7.2 ± 1.5 nmol min^-1 g^-1). Administration of radiolabelled choline at low (5 μM) and high (100 μM) concentrations, followed by microwave fixation 60 s later and chloroform-methanol-water separations of the homogenized brain did not suggest a relationship between concentration and the appearance of label in lipid or aqueous fractions as observed in another in-vitro study elaborating two-component kinetics of choline uptake. It was observed that 60s after carotid injection 12–14% of the radiolabel in the ipsilateral cortex was found in the chloroform-soluble fraction. Hemicholinium-3 (K_i= 111 μM), dimethylaminoethanol (K_i= 42 μM), tetraethyl ammonium chloride, tetramethyl ammonium chloride, 2-hydroxyethyl triethylammonium iodide, carnitine, normal rat serum, and to a lesser extent lithium and spermidine all inhibited choline uptake in the BBB. Unsubstituted ammonium chloride and imipramine did not inhibit choline uptake. No difference was observed in blood-brain barrier choline uptake of unanesthetised, carotid artery-catheterized animals, and comparable sodium pentobarbital-anesthetized controls.     

Measurement of inorganic pyrophosphate in biological fluids and bone tissues

A simple and rapid technique of measuring pyrophosphates in plasma, urine, and bone tissue is described, using the UDPG-pyrophosphorylase reaction and fluorimetrical determination of NADPH formed in a combined system of phosphorylation and reduction. This very specific method avoids the separation of Pi² by column chromatography, and its very great sensitivity enables measurement on a final sample corresponding to 0.2 ml of plasma, with a precision of about 3%.The mean plasma PPi concentration (±SE of mean) was 3.53 ± 0.19 μmoles/liter (SE=0.93), or 0.207 ± 0.006 mg Pi/liter. The normal range for this population (99% confidence limit) is therefore between 1.10 and 5.90 μmoles/liter or 0.068 – 0.366 mg Pi/liter. Analysis of the variation of the duplicate measurements shows a very small divergence of 3.4% or ±0.12 μm.Normal 24 hr urinary excretion is 53.0 ± 6.8 μmoles with 99% confidence limits of 10.0 and 96.0 μmoles.The average PPi concentration in calvaria of 20 10-day-old rats is 3.685 ± 0.16 nmoles/mg fresh weight.     

Perinatal changes in avian muscle: Implications from ultrastructure for the development of endothermy

Endothermic heat production and the capacity to shiver develop soon after hatching in birds, permitting chicks to regulate their body temperature. Physiological studies have not clearly identified the developmental events causing this change in function. Here, we use electron microscopy to examine the development of structures involved in muscle activation, contraction, and metabolism coincident with the development of shivering thermogenesis. A stereological study was used to compare the ultrastructure of chicken iliofibularis before endothermic heat production was present (24 h before hatching) and 120 h later, when the iliofibularis had substantial capacity for shivering. Profound increases were found in the t-tubule system and terminal cisternae, mitochondrial cristae, and lipids. The number of triadic profiles increased 3.8-fold (7.6 ± 1.31/100 μm² to 28.5 ± 2.90/100 μm² fiber area). The surface area of cristae per mitochondrial volume doubled (12.0 ± 1.50 pm2/pm3 to 25.7 ± 1.84 μm²/μm³). Lipid droplets were rare in the iliofibularis of embryos about to hatch, but accounted for 4.4% of the muscle fiber volume in day 4 birds. We suggest that these ultrastructural changes more fully activate the iliofibularis, allow it to produce more heat both from calcium pumping and from contraction, and increase its endurance, thus permitting the muscle to be effective in thermogenesis. © 1995 Wiley-Liss, Inc.     

Trypanosoma congolense: calf erythrocyte survival.

Hereford calves infected with Trypanosoma congolense developed an anemia which was most severe 10 weeks after infection when packed cell volumes (PCV) averaged 21.1 ± 2.5% (±2 SE) as compared to 33.1 ± 2.1% for controls. At the termination of the study, at 28 weeks postinfection PCVs of infected animals had risen to 27.5 ± 1.0% as compared to 34.0 ± 1.7% for controls. In parallel with PCVs the apparent half-lives of ⁵¹Cr-labeled erythrocytes were reduced as early as the first 2 weeks postinfection. The greatest difference in erythrocyte half-lives between infected and control animals was found during the fourth to sixth weeks of infection when volumes for infected animals averaged 128 ± 46 hr as compared to 321 ± 30 hr for controls. During this period the parasitemia was at its highest level. At 28 weeks postinfection the average apparent half-life of infected animals was 243 ± 43 hr compared with 304 ± 11 hr for controls. No differences were observed in gastrointestinal loss of ⁵¹Cr between infected and control animals; however, urinary excretion of isotope was greatly increased in infected animals when compared to controls. No significant changes in total blood volumes were observed between infected and control animals but total plasma volumes increased and total erythrocyte volumes decreased significantly in infected animals.     

An ultrastructural and developmental analysis of the corpus allatum of juvenile hormone deficient mutants ofDrosophila melanogaster

Summary The ultrastructure of the corpus allatum of theapterous mutantsap ⁴ andap ^56f ofDrosophila melanogaster during larval-pupal-adult metamorphosis and adult life was correlated with the gland's ability to synthesize juvenile hormone in vitro. During the early wandering period of the third instar of both mutants, a high concentration of smooth endoplasmic reticulum, mitochondria and mitochondrion-scalariform junction complexes are typical features of an active corpus allatum cell. Juvenile hormone biosynthesis by the glands is high at that time and, in fact, only slightly lower than that of wild type glands. In contrast to the wild type gland, the cells of the pupal and pharate adult corpus allatum of both mutants contains highly electron dense mitochondria with tubular cristae but no whorls of smooth endoplasmic reticulum nor glycogen clusters. The frequency and size of the lipid droplets, putatives depots of the juvenile hormone precursors, in cells of theap ^56f gland is a function of the insect's age, but both are lower than in wild type gland cells. Juvenile hormone biosynthesis by both mutant glands remains at the basal level when compared to increased synthesis by the wild type gland. The frequency and density of lipid droplets in cells of theap ⁴ corpus allatum are much lower than in theap ^56f glands. During adult life, the ultrastructural profile of theap ^56f corpus allatum is similar to that of the wild type gland although the in vitro production of juvenile hormone by the former is much lower than that of the wild type gland. The ultrastructural features of the adult corpus allatum ofap ⁴ homozygotes reveal precocious degeneration and support the view that this non-vitellogenic mutant is a juvenile hormone deficient mutation.