The influence of the thylakoid membrane surface properties on the distribution of ions in chloroplasts.

Thylakoid membranes isolated from peas have been subjected to ionic analyses using the technique of neutron activation. This has allowed the analyses of K+, Na+, Mg2+, Ca2+ and Cl- to be measured simultaneously on the same sample. By varying the ionic composition of the suspending medium it has been shown that these chloroplast membranes have no obvious chemical specificity for the inorganic cations studied and that the major controlling factor is the electrostatic neutralization of the surface negative charges. In agreement with the Gouy-Chapman theory and for the conditions used, divalent cations were preferentially attracted to the membrane surface. This finding, together with the ionic analysis of the unwashed thylakoids and of isolated intact chloroplasts, indicated that the major physiological surface cation is Mg2+ and that K+ is probably the main inorganic cation of the stroma. This conclusion is discussed in terms of counterion movement in response to light induced proton pumping at the thylakoid membrane.

Nonradioisotopic method for measuring iron absorption from a Gambian meal.

Iron absorption from a typical Gambian meal of rice with groundnut sauce was measured by the fecal balance technique in nonanemic adult males with 58Fe as an extrinsic label and neutron activation analysis and compared with ferrous sulphate. Gambian men had a higher efficiency of absorption than UK volunteers but the availability of the food Fe was approximately half that of ferrous sulphate in both groups of subjects.

Iron and zinc absorption in human subjects from a mixed meal of extruded and nonextruded wheat bran and flour.

The effect of extrusion cooking of a bran-flour mixture on iron and zinc retention was measured in normal adults. The stable isotopes 58Fe (1.253 mg) and 67Zn (5.13 mg) were administered with 40 g nonextruded or extruded cereal with milk and isotopic retention was measured from fecal excretion over the next 4-7 d by neutron-activation analysis (Fe) and fast-atom-bombardment mass spectrometry (Zn). 58Fe retention was 15.1 +/- 2.4% (means +/- SEM) with the nonextruded meal and 16.5 +/- 2.7% with the extruded meal. 67Zn retention was 18.9 +/- 1.7% with the nonextruded meal and 18.3 +/- 1.5% with the extruded meal. Extrusion cooking had no effect on 58Fe or 67Zn retention.

Metal ion release after total hip replacement.

The concentration of cobalt (Co), chromium (Cr), molybdenum (Mo), nickel (Ni), iron (Fe) and zinc (Zn) was measured using neutron activation analysis in tissue taken from an 81 year old female at necropsy. The patient had bilateral cobalt chromium molybdenum (Co-Cr-Mo) total hip replacements: one, a metal-on-metal had been in place for 14 years, the other a metal-on-plastic for 5.5 years. Although the metal-on-metal side had become painful, the patient remained active until she died. The measurements indicated that the concentrations of Co and Cr in the lung, kidney, liver and spleen were up to fifty times "Standard Man" values. High values occurred also in the urine and in the hair. The tissue adjacent to the metal-on-metal joint was heavily laden with metal wear debris, whereas that adjacent to the metal-on-plastic joint was relatively uncontaminated. The concentration varied with distance from the implant. Co predominated in the urine, whereas Cr predominated near the implants. The existence of such high levels, especially in the organs, is a possible cause for concern.

An application of neutron activation analysis to small biological samples: simultaneous determination of thirty elements in rat brain regions.

Thirty elements in 7 rat brain regions were determined by instrumental neutron activation analysis (INAA). The samples were irradiated by thermal neutrons using 3 different sets of conditions, depending on the nuclear characteristics of the elements. Analysis of the resulting radionuclides was by gamma-ray spectrometry using a high resolution Ge(Li) detector and Nuclear Data 6600 multichannel analyzer, which was fully computerized to give quantitative results for the gamma-ray spectra. This paper demonstrates the use of INAA for small biological samples and to show its potential elements, 7 rat brain regions are listed. It is interesting to note that certain elements, e.g. fluorine and potassium showed high and low regional differences respectively, and hypothalamus and hippocampus had higher elemental concentrations than other brain regions. However, since this paper is essentially an analytical one, no attempt is made to assess these data, which are preliminary, and the possible functional role of these elements will be discussed elsewhere.

Manganese mineral interactions in brain.

Manganese (Mn) is an essential mineral but is toxic when taken in excess. However, whether its interactions with other minerals in organs and cells are involved in mechanisms underlying Mn toxicity is poorly understood. We designed a developmental rat model of chronic Mn treatment (Group A: 1 mg MnCl2.4H2O per ml of drinking water; Group B: 10 mg MnCl2.4H2O per ml of drinking water; Group C: 20 mg MnCl2.4H2O per ml of drinking water; Control Group given water without manganese addition). Employing the model and instrumental neutron activation analysis, we investigated two hypotheses: (i) chronic manganese treatment alters the brain regional distribution of manganese and this altered manganese distribution also leads to region-specific changes of other metals; (ii) chronic manganese treatment induces differential changes in subcellular distributions of metals and electrolytes. In the treated rats, brain Mn level showed dose-related increases, the most pronounced being noted in striatum, hypothalamus, and hippocampus: these increases also led to alterations in regional distribution pattern of Mn. In the treated rats, Fe level was increased in hypothalamus, cerebellum, hippocampus, pons and medulla, and striatum. Cu level was increased in pons and medulla, hippocampus, midbrain, and striatum. Se level was increased in cerebellum, striatum, midbrain, hypothalamus, and pons and medulla. Zn level was increased in hypothalamus and striatum. Ca level was increased in midbrain but decreased in cerebellum; however, Mg and Al levels were not markedly affected. In brains of Mn-treated rats, Mn levels in subcellular fractions were all increased, being especially marked in nuclei, mitochondria, and synaptosomes; the subcellular distributions of Fe, Cu, Zn, and Mg were differentially altered although those of Al and Ca were minimally affected. These results are consistent with our hypotheses and may have implications in manganese neurotoxicity. The cellular and molecular mechanisms underlying manganese-mineral interactions in brain are still poorly defined and merit further investigation.

Loss of particulate contaminants from plant canopies under wet and dry conditions.

There is a requirement for data describing the loss with time of particulate contamination from plant canopies. Measurements were made of the loss rates of monodispersed silica spheres (three sizes, with Mass Median Aerodynamic Diameters (MMADs) 1.9, 5.3 and 8.4 microm) from wheat (Triticum aestivum) and broad bean (Vicia faba) canopies. The spheres were labelled with tracers detectable by Instrumental Neutron Activation Analysis (INAA). Canopies were contaminated under realistic turbulence conditions in a wind tunnel, then removed to sheltered and exposed field sites or to a glasshouse containing a rain simulator. Samples were taken periodically, and the level of contamination per plant determined by INAA. Statistical analysis of the resulting data suggested an offset exponential loss model, with a residue of deposit that is not lost over time. Loss half-lives in the order of 1-2 days were obtained for an exposed wheat crop and 3-4 days for a partially sheltered wheat crop, with permanent residues of initial deposit for the exposed crop of 4-8%, and for the partially sheltered crop of 22-52%. A broad bean crop under glasshouse conditions showed loss half-lives of 0.5-1.5 days with residues of 22-26% initial contamination. A double exponential loss model also fitted the data well in some cases, and it is possible that a slow loss of the residual deposit occurs, being masked by noise in the current data set.

Studies on iron availability in man, using stable isotope techniques.

1. Iron absorption from 10 mg Fe (as ferrous sulphate), labelled with 1.3 mg 58Fe, was measured in fasting, non-anaemic adult subjects by the faecal-balance technique. The measurement was performed twice, each subject being given, in random order, either 50 mg Fe or a placebo 18 h before the 58Fe-labelled FeSO4. 2. The 50 mg Fe load significantly reduced Fe absorption the following day (P less than 0.01), from a mean of 35.4 (SEM 4.6)% to 29.0 (SEM 5.1)%. This points to the importance of strict dietary control during Fe-absorption studies to eliminate bias in results. 3. In a separate study, the feasibility of using 58Fe-enrichment of erythrocytes, measured by neutron activation analysis (NAA), 10 d after a meal labelled with 0.69 mg 58Fe as an index of Fe absorption was examined. The levels of 58Fe in the blood were detectable by NAA. Regression analysis showed a significant relation between 58Fe-enrichment of blood and 58Fe absorption, calculated as the difference between intake and faecal excretion (R 0.59, P less than 0.05).

Iron absorption from a malted cocoa drink fortified with ferric orthophosphate using the stable isotope 58Fe as an extrinsic label.

The potential use of an extrinsic label to measure iron absorption from a ferric orthophosphate-fortified malted cocoa drink was examined by measuring the solubility of the FePO4 in 0.1 M-hydrochloric acid. The validity of using the stable isotope 58Fe as an extrinsic label was tested by comparing Fe absorption by rats from wheat flour extrinsically-labelled with 58Fe or 59Fe. Fe absorption from a malted cocoa drink (20 g powder made up with hot water) fortified with FePO4 (0.5 mg Fe/g powder) was measured in human subjects using 58Fe as an extrinsic label. Absorption was calculated by measuring unabsorbed 58Fe in faeces. Absorptions of Fe from the drink fortified with either FePO4 or ferrous sulphate were compared. The effect of the addition of ascorbic acid to the drink (1 mg/g powder) on Fe availability was also examined. The effect of fasting on Fe absorption from the drink was determined in rats by giving the drink to fasting animals or shortly after they had consumed a small meal. The FePO4 was totally soluble in 0.1 M-HCl and there were no differences in absorption between 58Fe- and 59Fe-labelled wheat flour. In the human experiment the proportion of Fe absorbed from the drink plus FePO4 and ascorbic acid was (mean with SE) 0.25 (0.02), from the drink plus FePO4 0.24 (0.02) and from the drink plus FeSO4 0.23 (0.03). Fasting had a significant effect on Fe availability; rats given the drink shortly after a small meal absorbed less than half as much Fe as those given the drink on a fasted stomach. It was concluded that the FePO4 used to fortify the malted cocoa drink was as well absorbed as FeSO4 but that the high levels of absorption were a reflection of the fasting condition of the subjects. The level of ascorbic acid was not great enough to enhance the availability of the FePO4 any further.

Neurochemical changes in rats chronically treated with a high concentration of manganese chloride.

Several neurochemical parameters were studied in brain regions of rats chronically treated with a high concentration of manganese chloride (20 mg MnCl2.4H2O per ml. of drinking water) throughout development until adulthood. Large increases in Mn accumulation were found in all brain regions (hypothalamus, +530%; striatum, +479%; other regions, +152 to +250%) of Mn-treated adult rats. In these animals, Ca levels were decreased (-20 to -46%) in cerebellum, hypothalamus, and cerebral cortex but were increased (+186%) in midbrain. Mg levels were decreased (-12 to -32%) in pons and medulla, midbrain, and cerebellum. Fe levels were increased (+95%) in striatum but were decreased (-28%) in cerebral cortex. Cu levels were increased (+43 to +100%) in pons and medulla and striatum but Zn levels were decreased (-30%) in pons and medulla. Na levels were increased (+22%) in striatum but those of K and Cl remained unchanged. Type A monoamine oxidase activities were decreased (-13 to -16%) in midbrain, striatum, and cerebral cortex, but type B monoamine oxidase activities decreased (-13%) only in hypothalamus. Acetylcholinesterase activities were increased (+20 to +22%) in striatum and cerebellum. The results are consistent with our hypothesis that chronic manganese encephalopathy not only affects brain metabolism of Mn but also that of other metals.

Changes in brain regional manganese and magnesium levels during postnatal development: modulations by chronic manganese administration.

Manganese (Mn) and magnesium (Mg) levels in hypothalamus, cerebellum, pons and medulla, striatum, midbrain, and cerebral cortex of control and Mn-treated (10 mg MnCl2.4H2O per ml of drinking water) rats during postnatal development were studied using instrumental neutron activation analysis. The age-dependent Mn accumulation showed regional variation: at day 5, this accumulation was most marked in striatum (12.05 micrograms/g wet weight) but least marked in cerebral cortex (0.85 micrograms/g wet weight). By day 10, pons and medulla, and hypothalamus were regions with, respectively, the highest (4.73 micrograms/g wet weight) and the lowest (0.52 micrograms/g wet weight) Mn levels. By contrast, brain regional Mn variations were less pronounced in weanling and adult rats. The age-dependent Mg accumulation showed regional variation at day 5, being most marked in pons and medulla (720 micrograms/g wet weight) and least marked in cerebral cortex (295 micrograms/g wet weight). Mg levels in all regions decreased after day 5; by day 120, only Mg level in cerebral cortex was lower than levels in other regions (the latter being very similar). In general, the age-related decreases in Mn and Mg levels paralleled the decreases in water content and increases in tissue weight, suggesting that the maturation of the blood-brain barrier may play important role(s) in brain Mn and Mg homeostasis. Chronic Mn-treatment from conception onwards altered the regional Mn and Mg distribution patterns during development. Our results are consistent with the hypothesis that chronic Mn toxicity exerts modulatory effects on brain regional metabolism and homeostasis of Mn and other metals during development.

Effects of chronic manganese treatment on rat brain regional sodium-potassium-activated and magnesium-activated adenosine triphosphatase activities during development.

The effects of chronic manganese (Mn) treatments (1 and 10 mg MnCl2.4H2O per ml of drinking water) from conception onwards on brain regional development of sodium-potassium-activated and magnesium-activated adenosine triphosphatases (Na-K-ATPase and Mg-ATPase) were studied. The activities of these enzymes were determined in hypothalamus, cerebellum, pons and medulla, striatum, midbrain and cerebral cortex (which included the hippocampus) of Mn-treated and age-matched control rats at 5 postnatal ages. Both ATPase activities doubled in most brain regions between day 5 and day 20 postnatal. In pons and medulla, striatum, midbrain, and cerebral cortex, adult levels of both enzymatic activities were attained by day 20 postnatal. Na-K-ATPase activities transiently increased in the midbrain (+25%) at day 12 with the lower Mn dose and in the cerebral cortex (+31%) at day 20 with the higher Mn dose. With the higher Mn dose only, Mg-ATPase activities were increased in the hypothalamus (+20%) at day 12 and in the pons and medulla (+22%) at day 20 but were decreased in the pons and medulla (-20%) at day 60. Thus, only transient changes in enzymatic activities were observed despite dose-dependent increases in the brain levels of Mn resulting from the Mn treatment. A hypothesis regarding the role of early but transient changes in brain metabolism in the pathogenesis of the initial psychotic symptoms in Mn intoxication was proposed and discussed in relation to several other observations of a similar nature.

Riboflavin deficiency and iron absorption in adult Gambian men.

Iron absorption from 3.38 mg 58Fe was measured in riboflavin-deficient Gambian men with haemoglobin (Hb) less than 11.5 g/dl before and after oral riboflavin therapy, and the results compared with a group not receiving riboflavin. Riboflavin status (as determined by erythrocyte glutathione reductase activation coefficient) and Hb increased in teh riboflavin-supplemented but not placebo group. Plasma ferritin levels were low and did not change in either group. There was very wide variation in percentage iron absorption between individuals and also within single individuals on two separate occasions but no measurable change with riboflavin supplementation. The results of the study indicate that the efficiency of iron utilization is impaired in riboflavin deficiency, but that iron absorption is unaffected.

Lactoferrin and iron absorption in newborn infants.

Results from experiments in this laboratory using 59Fe suggest that bovine lactoferrin (Lf) has no effect on iron absorption in rats. A study was therefore carried out in newborn infants to measure the effects of Lf on iron retention. Bovine Lf was labeled with the stable isotope 58Fe and fed to 7-day-old infants in a standard milk formula. Iron retention was estimated by measuring the unabsorbed 58Fe excreted in the feces during the following 3 days using neutron activation analysis. The results were compared with those obtained from a group of infants fed a similar level of iron as ferric chloride, labeled with 58Fe, together with 30 mg ascorbic acid. There was a very wide variation in percent iron retention amongst the infants but no overall difference between the Lf and ferric chloride groups. This confirms the previous findings in rats that Lf does not influence the availability of nonheme iron.

The effect of moderately increased intakes of complex carbohydrates (cereals, vegetables and fruit) for 12 weeks on iron and zinc metabolism.

Fifteen adult women were given diets in which the intake of complex carbohydrates was increased from 20 to 30 g over a 12 week period. Metabolic balances were carried out, iron and zinc absorption tests performed using stable isotopes, and Fe and Zn status monitored. Although effects on bowel function were observed, the changed diet had no influence on any of the previously described variables. It was concluded that a moderate increase in cereals, fruit and vegetables did not have an adverse effect on Fe or Zn nutrition.