Tegumental glucose permeability in male and female Schistosoma mansoni

Tegumental hexose transporters have been kinetically characterized in mated and separated male and female Schistosoma mansoni 8-12 wk postinfection. Significant gender-specific differences in Km and Vmax were observed. In mated males, the estimated constants (mean +/- SE) were: Km = 0.63 +/- 0.31 mM, Vmax = 0.93 +/- 0.44 nmol/mg worm water/min, and the Kd = 0.25 +/- 0.09 microliter/mg worm water/min. In mated females the kinetics were: Km = 0.99 +/- 0.40 mM, Vmax = 1.22 +/- 0.42 nmol/mg worm water/min, and Kd = 0.60 +/- 0.14 microliter/mg worm water/min. The influx of 2-deoxy-D-glucose and 3-O-methylglucose has been similarly characterized; these analogs share the same glucose transporter in male and female schistosomes. 2-Deoxy-D-glucose has a higher affinity, and 3-O-methylglucose a lower affinity, than does glucose. Because mated male schistosomes supply glucose to female partners, similarities between the free glucose concentration of the male and the affinity of the transporter determined for mated female schistosomes suggest that male-to-female transfer may be a potentially rate-limiting step in glucose utilization by the female. Permeability x surface are (PS) products and Vmax/Km ratios were significantly elevated in mated schistosomes, suggesting that the transporter is primarily localized to the dorsal surface of the male. Gender- and mating-specific analyses of PS products indicate that tegumental permeability to glucose is significantly increased in mated schistosomes, and compares very favorably to that of the host liver.     

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.     

Reduction in Brain Glucose Utilization Rate after Tryptophol (3-Indole Ethanol) Treatment

Abstract: 3-Indole ethanol has been recently identified as the hypnotic agent in trypanosomal sleeping sickness, and because it is formed in vivo after ethanol or disulfiram treatment, it is also associated with the study of alcoholism. When administered intraperitoneally to rats (250 mg/kg) tryptophol induced a sleep-like state that lasted less than an hour (no righting reflex was apparent 2 min after injection, but it returned at 11 min in bovine serum albumin solution, and 47 min in 40% ethanol solution). In ethanol solutions, tryptophol reduced brain cortical glucose utilization by 55% to the basal brain metabolic rate, and this effect lasted less than 1 h. Synergistic effects of tryptophol and ethanol were suggested by the observation that in albumin solution, tryptophol reduced brain glucose utilization by 35%, but a normal rate was not observed until 2 h postinjection.     

Contraceptive Use and Pregnancy Outcomes among Opioid Drug-Using Women: A Retrospective Cohort Study

Objective

The contraceptive needs of illicit opioid users differ from non-drug users but are poorly understood. The aim of this study was to describe contraceptive use and pregnancy outcomes in opioid-using women, and to examine their association with a range of risk factors.

Method

This retrospective cohort study used UK general practice records, Treatment Outcomes Profile and National Drug Treatment Monitoring System data, and a nested data validation exercise. A cohort of 376 women aged 20–61 years were in active treatment for opioid addiction in October 2010 at two specialised primary care practices in North-East England. Outcomes were age-adjusted prevalence estimates for contraceptive use and pregnancy outcomes in users of illicit opioids. The association between lifestyle-related risk factors and contraception was explored.

Results

Drug-using women made lower use of planned (non-condom) contraception (24% vs 50%, p<0.001), had more frequent pregnancy terminations (0.46 vs. 0.025, p = 0.004) and higher annual incidence of chlamydia (1.1% vs. 0.33%, p<0.001), when compared with age-matched population data. Specifically, there was low use of oral contraceptives (4% vs. 25%, p<0.001), IUCD (1% vs. 6%, p<0.001), and sterilisation (7% vs. 6%, p = 0.053), but higher rates of injectable contraceptives (6% vs. 3%, p = 0.003). A total of 64% of children aged <16 years born to this group did not live with their mother. No individual risk factor (such as sex-working) significantly explained the lower use or type of non-condom contraception.

Conclusions

This is the first study to describe planned contraceptive use among drug-users, as well as the association with a range of risk factors and pregnancy outcomes. The low uptake of planned contraception, set against high rates of terminations and sexually transmitted disease demonstrates the urgent clinical need to improve contraceptive services, informed by qualitative work to explore the values and beliefs influencing low contraceptive uptake.     

A national survey of airborne pollen and grass flowering in New Zealand,with implications for respiratory disorder

Airborne pollen and spore levels were monitored at seven sites in New Zealand using the Intermittent Cycling Rotorod sampler during the summer of 1988/1989. Grasses formed the major component of atmospheric pollen levels during spring and summer at every locality. Peak levels of grass and total pollen occurred during December or late November, with a slight latitudinal lag apparent at the more southern sites. Highest levels were recorded at the smaller rural centres of Gore and Kaikohe and the lowest at the larger urban centres of Auckland, Christchurch and Wellington. We make a first approximation of the likely risk to hayfever and allergic asthma patients at each of the seven centres. For example, significantly higher grass pollen levels were experienced at Kaikohe on 44% and 65% of days during November and December, compared with just 15% and 8% at Auckland. By recording the flowering seasons of the principal allergenic grass species at each locality, we determined the potentially allergenic grasses contributing to peak pollen levels, the most ubiquitous being tall fescue (Festuca arundinacea Schreb.), ryegrass (Lolium perenne L.,L. multiflorum Lam.), cocksfoot (Dactylis glomerata L.), Yorkshire fog (Holcus lanatus L.) and sweet vernal (Anthoxanthum odoratum L.). Corresponding author. Deceased.     

Regional modulations in tegumental glucose transporter kinetics in the rat tapeworm

Comparisons of glucose transporter kinetics in 8-day (Km = 0.34 mM, Vmax = 14 nmole.min-1.g-1), 10-day (Km = 0.46 mM, Vmax = 18 nmole.min-1.g-1), the first quartile of 17-day (Km = 0.51 mM, Vmax = 21 nmole.min-1.g-1), and the first quartile of 32-day (Km = 0.33 mM, Vmax = 39 nmole.min-1.g-1) rat tapeworms (Hymenolepis diminuta) suggest maximal velocities may vary with age. A gradient in glucose transporter density is suggested in the rat tapeworm by changes in the estimated transporter Vmax in the first through fourth quartiles. Alterations in the physiological efficiency (as indicated by the Vmax/Km ratio) and permeability (indicated by the unsaturated permeability-area product) of the glucose transporter were determined to be significantly greater in the first quartile than in other quartiles of 17-day hymenolepids. A similar trend was apparent in older (32-day) worms. In tapeworms maintained for 30 min in glucose-free medium, maximal velocities were highest in the anterior (first) quartile, and reductions were seen in successive second, third, and fourth quartiles. When worms were maintained in a medium containing 11 mM glucose, maximal velocities were about twofold greater, but the Vmax increased in each successive quartile. The apparent half-saturation constants, which indicate that concentration of external glucose at which half of the glucose transporter proteins are occupied, are reduced approximately 50% in tapeworms maintained in glucose-free medium. These studies demonstrate that regional differences exist in the glucose transporter of the rat tapeworm, analogous to the intestinal glucose gradient. Furthermore, substrate-induced modulations in the transporter may also exhibit independent regional variability.     

Expression of α-amylase and other gibberellin-regulated genes in aleurone tissue of developing wheat grains

Aleurone tissue from freshly harvested immature wheat grains (Triticum aestivum L. cv. Sappo) which is normally unresponsive to gibberellic acid can be made responsive by subjecting the tissue to a pre-incubation treatment in a simple buffered medium prior to the addition of the growth substance. The effectiveness of this treatment is dependent on grain age, with grains less than 15–20 days post anthesis failing to become converted to a responsive state whilst tissue from grains older than this become increasingly susceptible. Tissue from grains of a certain age (approx. 25–28 days post anthesis) produce small amounts of -amylase following this treatment even in the absence of exogenously applied growth substance. Using different ³²-labelled complementary-DNA probes for -amylase in wheat it was demonstrated that the failure of freshly harvested tissue to produce -amylase was correlated with the absence of the appropriate mRNA species. Inability to accumulate -amylase mRNA in response to gibberellic acid was removed by the pre-iccubation treatment and also by enforced drying. The gibberellin-regulated expression of other unidentified genes also responds to pre-incubation or drying. Induction of gibberellin-responsiveness in immature aleurone cells did not extend to the secretion of acid phosphatase, protease and ribonuclease.Abbreviations cDNA complementary DNA - dpa days post anthesis - GA gibberellin - GA₃ gibberellic acid     

Nutrient transport and the blood-brain barrier in developing animals

Structural alterations in the development of the blood-brain barrier (BBB) can be seen in capillary profiles from the rat cortex. The neonatal luminal membrane is amplified with irregular folds, a possible adaptation to reduced cerebral blood flow rates. By 21 days the capillaries have resolved to a smooth-surfaced, adult-like appearance. Developmental alterations in the basement membrane, tight junctions, capillary seams, Golgi, pinocytotic vesicles, and cytoplasmic thickness are observed. Two studies have addressed developmental modulations in BBB polarity; both indicate that brain-to-blood transport mechanisms that were inoperative in the early neonatal rat become functional in weanlings. Six of the seven major independent BBB nutrient transport systems that regulate plasma-to-brain uptake have been kinetically characterized in the newborn rabbit, and comparisons have been made in the weanling (28-day-old) rabbit. All of these saturable transport systems are operative at birth, which suggests that the immature rabbit has a mature BBB with respect to regulation of nutrients. Purine base permeability, affinity, and uptake velocities are virtually unchanged during postnatal development. Subtle alterations in amino acid and amine transport were suggested by the lower-affinity (high-capacity) transport mechanisms characterized in the newborn as compared to the 28-day-old BBB. Under conditions of elevated plasma levels (typical of the neonate), these higher-capacity mechanisms would facilitate a relative increase in metabolite influx to the developing brain. Significant differences in kinetics were also observed for the monocarboxylic acid and hexose transport systems in the absence of developmental changes in permeability times surface area products. A low-affinity, high-capacity monocarboxylic acid transport system operates at birth. It supplies the developing brain with increased quantities of ketone bodies, but is seen as a high-affinity, low-capacity mechanism in the 28-day-old rabbit. Concomitantly, the higher-affinity glucose carrier defined in newborn rabbits modulates, and by 28 days becomes a lower-affinity, high-capacity mechanism capable of delivering about 2 mumol X min-1 X g-1 of glucose to the (anesthetized) brain.     

Cupular secretion by Xenopus laevis line organs: autoradiographic evidence for incorporation of 3H-glucose and 35S-sulfate

Autoradiographic evidence for incorporation of 3H-glucose and 35S-sulfate into the cupulae of Xenopus laevis (African clawed toad) lateral line organs was obtained after injection into the dorsal lymph sacs of adult animals. Time intervals of 15 minutes to 4 hours after administration of these labeled metabolic precursors were used to examine the time course of the apparent mechanism of growth of the cupulae. Our results suggest that the two layers of accessory cells (the sustentacular cells and inner layer of mantle cells), concentrically arranged around the organ's central sensory (hair) cells, elaborate distinct cupular components. Sustentacular cells, immediately adjacent to the sensory cells, appear to produce and extrude at their exposed apices a cupular "core" substance labeled by 3H-glucose, but not by 35S-sulfate. The layer of inner mantle cells, external to the sustentacular cells, was labeled by both precursors and is spatially situated to secrete a cupular sheath enclosing the cupular core. Ultrastructural differences between the secretory products within the two cell types were marked. Electron microscopic autoradiography of toads killed 4 hours after 3H-glucose injection showed that silver grains were associated with accumulations of the respective secretory products in sustentacular and inner mantle cells, and label was found over the cupular trough area, where the bases of the cupulae are attached. These results suggest that the cupular core and sheath may both contain mucopolysaccharide, and the sheath, a sulfated mucopolysaccharide.     

Newborn Rabbit Blood–Brain Barrier Is Selectively Permeable and Differs Substantially from the Adult

Abstract: Examination of blood-brain barrier (BBB) function by the intracarotid injection technique has been utilized in studies of newborn (6–30 h) and adult rabbits. The exclusion of mannitol (mol. wt. 182), dextran (mol. wt. 60,000–90,000) and indium-bound EDTA indicate that the newborn BBB has restrictive properties similar to the adult. At birth, saturable, carrier-mediated transport mechanisms are present, regulating the entry of glucose, amino acids, organic acids, purines, nucleosides and choline. No difference in brain uptake of glucose was observed between adult and newborn, but considerably higher uptake rates for arginine, choline and adenine were seen in the newborn. In contrast to suggestions of an immature barrier in young animals, these studies indicate that a sophisticated, selective BBB is operative at birth. Furthermore, the specific selectivity and dramatic increases seen for certain metabolites imply a vital function in the newborn for these carrier systems.