Intestinal transfer of manganese: resemblance to and competition with calcium.

The effect of calcium, phosphate and the sugars lactose and sorbitol on the intestinal absorption of manganese were studied in adult male rats. Gastric gavage showed that lactose (100 mM or 200 mM) increased the hepatic retention of 54Mn, while phosphate decreased it. In situ ileal loop studies indicated that Mn absorption was normally complete in 30 min. Sorbitol had no effect on uptake during this period, but extended Mn absorption from 30 min to 120 min. Low concentrations of Mn (10 microM) did not alter the enhancing effect of lactose on calcium transport (10 mM), but the enhancing effect of lactose on Mn transport was blocked by this high calcium concentration. Intestinal alkaline phosphatase activity was rapidly stimulated by Mn. These similarities plus the competition between cations, especially calcium, suggest that a common mechanism exists in their intestinal transport.     

Fine structure of the midgut and Malpighian papillae in Campodea (Monocampa) quilisi Silvestri, 1932 (Hexapoda, Diplura) with special reference to the metal composition and physiological significance of midgut intracellular electron-dense granules

The fine structure of the midgut and the Malpighian papillae in Campodea (Monocampa) quilisi Silvestri, 1932 (Hexapoda, Diplura) specimens was described. We observed the presence of electron-dense granules (EDGs) in the midgut epithelial cells, similar in genesis, structure and aspect to the type A spherocrystals described in the midgut epithelium of Collembola and Diplopoda. Energy-dispersive X-ray microanalysis was used to detect the chemical composition of the granules and to relate it to the concentrations of some potential toxic heavy metals (Pb, Cu, Zn) in soil and litter. Chemical composition of the granules seems strongly influenced by the presence and bioavailability of heavy metals in the external environment. Specimens from a contaminated abandoned mining and smelting area (Colline Metallifere, southern Tuscany) were able to accumulate Fe, Mn, Zn, Pb and Cu in their midgut EDGs. In addition, we observed that C. (M.) quilisi was able to excrete the metal-containing granules into the external medium by the moulting of the intestinal epithelium. This confirms that the process of ionic retention of midgut cells is particularly significant in animals lacking Malpighian tubules.     

Quantitative energy dispersive X-ray microanalysis of eight elements in pancreatic endocrine and exocrine cells after cryo-fixation

Quantitative X-ray microanalysis of 8 elements was performed on ultrathin, freeze-dried sections of islets and pancreas pieces from non-inbredob/ob-mice. Diffusion of elements was reduced to a minimum by rapidly freezing the tissue samples between nitrogen-cooled polished copper surfaces and avoiding the use of chemical fixatives and stains. The ultrastructural morphology was adequately maintained to allow measurements on secretory granules, mitochondria, cell nuclei, and cytoplasm free of these organelles. The distribution of the various elements between cellular compartments was similar in islet -cells and exocrine pancreas cells. However, the insulin secretory granules were outstanding in exhibiting the highest concentrations of zinc and calcium. In comparison with cytoplasm in the -cells, the insulin granules accumulated calcium 2-fold and zinc as much as 40-fold. As no correlation could be made for endoplasmic reticulum in the cytoplasmic measurements areas, the true accumulations above cytosol are likely to be even higher.     

Cytochemical and X-ray microanalytical studies of calcium in the extending zone, particularly in the rhizodermis of Zea mays roots

Calcium was detected in CaCl (10 m M )-pretreated roots of Zea mays L. (cv. LG 11) by means of electron probe X-ray microanalysis with energy-dispersive spectrometry (EDS) using embedded samples, fixed by the antimonate staining procedure. A high level of calcium was found where large amounts of antimonate precipitates were observed by light or transmission electron microscopy. In the elongation zone, after 20 h in humid air following a 2 h CaCl₂ pretreatment, the level of calcium was higher in trichoblasts than in atrichoblasts. In these cells it was detected mainly in the walls and nucleus, and antimonate staining was observed in the walls. Abundant precipitates containing calcium were associated with the nucleus, vacuoles, mitochondria and endoplasmic reticulum of non-differentiated cells, whereas they were confined to the walls of these cells just after the CaCl₂ pretreatment. The involvement of calcium in the formation of root hairs is discussed.     

Role of intestinal mucus in crystal biogenesis: an electron-microscopical,diffraction and X-ray microanalytical study

Summary In the posterior intestine of the sea-water eel, mucus plays an important role in biocrystallization of calcium ions. By means of transmission and scanning electron microscopy associated with X-ray microanalysis and X-ray diffraction it has been possible to determine the role of mucous fibers as nucleation sites. Biocrystallization occurs in 2 steps: (1) Calcification of mucus. As soon as mucus is excreted in the intestinal lumen, it is loaded with calcium, as shown by lanthanum affinity and X-ray microanalysis on freeze-dried tissues. (2) Genesis of crystals. Needleshaped crystallites build up in coalescent spherites in the intestinal lumen near the microvilli. Genesis occurs as follows: (a) crystallite mineralization by nucleation in an organic matrix composed of glycoproteinaceous mucous fibers, followed by the appearance of spherites; (b) coalescence in spherites and association of spherites in rhombohedra; (c) extrusion of organic material during the final step of crystallization.     

Effect of silver nanoparticles on growth performance, metabolism and microbial profile of broiler chickens

This study evaluated the potential of silver nanoparticles (AgNano) as an antimicrobial growth-promoting supplement for broiler chickens. One hundred forty-four seven-day-old broiler chicks were distributed randomly to AgNano treatments at 0, 10 and 20 mg/kg (Control, Group AgNano10, and Group AgNano20, respectively) provided via the drinking water from day 7 to 36 post-hatching. Body weight and feed consumption were measured weekly. In addition, balance and respiration experiments were carried out to determine nitrogen (N) utilisation and energy retention. At days 22 and 36, blood samples and intestinal content were collected to evaluate the effects of AgNano on plasma concentration of immunoglobulins and the intestinal microflora, respectively. The provision of water solutions containing different concentrations of AgNano had no effect on postnatal growth performance and the energy metabolism of broiler chickens. However, in Group AgNano10 N intake (p = 0.05) and retention (p = 0.03) was increased, but N excretion and efficiency of utilisation was not affected. The populations of bacteria in the intestinal samples were not affected by AgNano supplementation. The concentration of immunoglobulin (IgG) in the blood plasma of broilers supplemented with AgNano decreased at day 36 (p = 0.012). The results demonstrated that AgNano affects N utilisation and plasma IgG concentration; however, it does not influence the microbial populations in the digestive tract, the energy metabolism and growth performance of chickens.     

Ultrastructural Localization of Calcium in the Presumptive Ectodermal Cells in Gastrulae of the Newt, Cynops pyrrhogaster, as Revealed by Cytochemistry and X-Ray Microanalysis

The ultrastructural localization of calcium in the presumptive ectodermal cells of gastrulae of the newt, Cynops pyrrhogaster , was examined by cytochemical methods and X-ray microanalysis (XMA). The cells were fixed with solutions that contained potassium oxalate, potassium ferricyanide and potassium pyroantimonate to preserve the localization of intracellular calcium in situ and for the analysis of electron density due to calcium. Electron-dense deposits associated with the localization of calcium were observed under the electron microscope. Specificially, pigment granules, round vacuoles, endoplasmic reticulum and mitochondria as well as the extracellular matrix were observed to contain calcium. In addition, XMA clearly demonstrated the localization of calcium in all of these electron-dense organelles and yolk granules.     

Ingestion of difructose anhydride III enhances absorption and retention of calcium in healthy men

We examined the effects of a nondigestible disaccharide difructose anhydride III (DFAIII) on calcium absorption and retention by means of a human balance study of single-blind crossover design. Twelve healthy male subjects ingested 250 mg of shell powder as calcium carbonate (corresponding to 100 mg of calcium) with or without 1.0 g DFAIII three times a day for 13 d. In the last 4 d as a balance period, all urine and feces were collected and evaluated for calcium excretion. The apparent calcium absorption (mg/d) and rate of absorption (%) were higher, and those of retention were much higher, in the DFAIII group than in the control group. Furthermore, serum osteocalcin increased after the experimental period in the DFAIII group but not in the control group. These results indicate that DFAIII ingestion enhances intestinal calcium absorption, which might be beneficial for bone metabolism.     

Effects of watershed liming on the soil chemistry of Woods Lake,New York

The effects of watershed liming on the exchange complex of a forest soil were investigated at Woods Lake, in the west-central Adirondack Park, New York. Attempts to neutralize lake acidity via direct application of calcite during the 1980"s were short-lived due to a short hydraulic retention time. The Experimental Watershed Liming Study (EWLS) was initiated to investigate watershed base addition as a potentially more long-term strategy for mitigation of lake acidity. In this paper we discuss the changes in the exchangeable soil complex which occurred in response to the calcite addition and attempt a mass balance for calcite applied to the watershed.An extensive sampling program was initiated for the watershed study. Soil samples were collected from pits prior to and in the two years following treatment to evaluate changes in soil chemistry. Calcite addition significantly altered the exchange complex in the organic horizon. Increases in pH caused deprotonation of soil organic matter and increases in cation exchange capacity, providing additional exchange sites for the retention of added calcium. Exchangeable acidity decreased to very low values, allowing the base saturation of upper organic horizons to increase to nearly 100 percent.Post-treatment sampling found that approximately 48 percent of the calcite remained undissolved in the soil"s Oe horizon two years later. Dissolution of the calcite was affected by field moisture conditions, with greater dissolution in wetter areas of the watershed. Mass balances calculated for calcium applied to the watershed suggest that only 4 percent of the calcium was removed through the lake outlet. Approximately 96 percent of the calcium applied remained within the watershed; as undissolved calcite, on soil exchange sites or stored in the vegetation, groundwater or surface waters of the watershed.     

The distribution of zinc, cadmium, lead and copper within the hepatopancreas of a woodlouse

The distribution of metals within the hepatopancreas of Oniscus asellus (Crustacea, Isopoda) from two uncontaminated sites, and two sites contaminated with zinc, cadmium and lead, has been studied by atomic absorption spectrophotometry, light microscopy, transmission and scanning electron microscopy and X-ray microanalysis. The hepatopancreas contains two types of intracellular granule. The first type, in the S cells, are spherical granules which contain copper, sulphur and calcium. In woodlice from contaminated sites, these ‘copper’ granules, also contain zinc, cadmium and lead. The second type, in the B cells, are flocculent deposits which contain iron. In woodlice from contaminated sites, these ‘iron’ granules also contain zinc and lead. Moribund woodlice from contaminated sites have large numbers of ‘copper’ and ‘iron’ granules in the hepatopancreas and a fine deposit of zinc and lead on the membranes of the cells. There are numerous microorganisms in close association with the microvillous border of the hepatopancreas of woodlice from all four sites. Within the microorganisms of Oniscus asellus from contaminated sites, there are deposits of material which contain zinc, lead, calcium and phosphorus ‘Copper’ and ‘iron’ granules could have evolved as storage sites for essential metals to be utilized when demand from the body exceeds uptake from the food. Woodlice in contaminated sites may be able to ‘detoxify’ potentially harmful amounts of essential and non-essential metals by storing them in a relatively insoluble form within these granules.     

向日葵柱头,花柱和珠孔中钙分布的超微细胞化学定位

用焦锑酸盐沉淀法对向日葵（ＨｅｌｉａｎｔｈｕｓａｎｎｕｕｓＬ．）授粉前后柱头、花柱和珠孔中的钙进行了超微细胞化学定位。同时还运用Ｘ射线能谱（ＥＤＸ）和波谱（ＷＤＸ）两种方法进行了Ｘ射线定性分析,证明了前法所得沉淀确系焦锑酸钙。观察表明,花粉萌发和花粉管生长所经的柱头接受面,花柱引导组织和珠孔引导组织中含钙较柱头、花柱和珠孔的其它部位明显地多。柱头乳突细胞的表面和花柱引导组织的胞间基质中、尤其胞间基质与细胞壁外层相接之处钙很密集。在珠孔外端引导组织中,以角质层为界,钙主要分布于其近珠柄侧。花粉管壁果胶质层中有相当多的钙。结合向日葵中已有的研究和其他文献,讨论了钙的分布与花粉管生长的关系     

X-ray microanalysis of the mineral components in the scales of an amoeba,Cochliopodium sp. (Testacea)

Summary Mineral components of the scales in an amoeba, Cochliopodium sp., were examined by energy-dispersive X-ray microanalysis. The precipitates in potassium antimonate-treated material detected calcium in the scales. Calcium was also clearly detected in freeze-substituted thin sections. Similar deposits of calcium antimonate were detected in scales in formation within vacuoles, and also in Golgi cisternae, Golgi vesicles and special granules near the nucleus. There were only minute amounts of magnesium and potassium. This suggests that calcium is the main mineral component of the scales and that it is added in the Golgi complex during scale formation.     

1 alpha, 25-Dihydroxy-vitamin D-3 regulates ATP-dependent calcium transport in basolateral plasma membranes of rat enterocytes

Basolateral plasma membrane vesicles of rat small intestinal epithelium accumulate calcium through an ATP-dependent pumping system. The activity of this system is highest in duodenum and decreases towards the ileum. This distribution along the intestinal tract is similar as the active calcium absorption capacity of intact intestinal epithelial segments. ATP-dependent calcium uptake in basolateral membrane vesicles from duodenum and ileum increased significantly after repletion of young vitamin D-3-deficient rats with 1 alpha,25-dihydroxy-vitamin D-3. Ca2+ -ATPase activity in duodenal basolateral membranes increased to the same extend as ATP-dependent calcium transport, but (Na+ + K+)-ATPase activity remained unaltered.     

Intestinal apoB synthesis, lipids, and lipoproteins in chylomicron retention disease

Chylomicron retention disease is characterized by fat malabsorption, hypocholesterolemia, normal fasting triglycerides, and marked intestinal steatosis despite the presence of both plasma and intestinal apoprotein B. The defect remains unknown but presumably involves the synthesis or secretion of chylomicrons. The present investigation examines this hypothesis by studying the biosynthesis of chylomicrons in cultured jejunal explants and by defining the quantitative and qualitative abnormalities of plasma lipids and of circulating lipoproteins. Following 2-3 years of a low fat diet supplemented with medium chain triglycerides, six patients with chylomicron retention disease had significantly higher triglyceride (TG) levels coupled with a decrease in both free (FC) and esterified cholesterol (EC) as well as in essential fatty acids and phospholipids (PL) when compared to healthy controls. The low total plasma cholesterol was largely accounted for by low levels of both low density (LDL) and high density lipoprotein (HDL) cholesterol. VLDL and LDL were characterized by a diminished percentage of CE with an increase of TG while HDL contained relatively more FC as well as PL and less CE. The diameter of VLDL was larger whereas those of LDL and HDL were smaller than in normal controls. Jejunal explants, when incubated with [14C]palmitate, were capable of normal biosynthesis of TG, diglycerides, PL, and CE. These lipids, however, except for PL, were retained in the tissue and could not be secreted into the culture medium. Incubation of intestinal biopsies with [3H]leucine and [14C]mannose resulted in normal protein synthesis and reduced glycosylation. The presence of intestinal apoB-48 was confirmed by immunoblot using 2D8 antibodies. These data suggest that the intestinal defect in this disease results from a disorder of the final assembly of chylomicrons or in the mechanism of their exocytosis.     

Phosphorylation of peptides derived from isolated soybean protein: effects on calcium binding, solubility and influx into Caco-2 cells

Calcium homeostasis in the human body is maintained primarily via the absorption of calcium through the intestine. In order to maintain an efficient absorption of calcium with minimal calcium loss due to the formation of calcium phosphate precipitates in the small intestinal lumen, we developed a calcium-binding mediator using peptides derived from isolated soybean protein (ISP). ISP was modified via tryptic digestion and chemical phosphorylation using sodium trimetaphosphate, thereby generating soybean phosphopeptides (SPP), and this was followed by conducting a binding reaction with calcium chloride. We have established an optimized procedure and reaction conditions for maximal phosphorylation and calcium binding. Consequently, the phosphorylation of soybean peptides resulted in considerable improvement in their calcium binding activities. Next, we demonstrated that SPP was able to render calcium ions resistant to precipitate formation with inorganic phosphates, which suggested the enhancement of calcium bioavailability. Finally, we noted that the addition of calcium-bound SPP induced an increase in cytosolic calcium concentration in the intestinal Caco-2 cells, due to an influx of calcium. These findings provide a new basis by which we may assess the possibility that SPP, as a potent calcium carrier, can be utilized in the prevention of poor absorption of dietary calcium in animals.     

Effects of chronic ethanol feeding and acetaldehyde metabolism on calcium transport by rat liver mitochondria

The prolonged feeding of ethanol to rats alters in vitro mitochondrial transport of calcium. Hepatic mitochondria isolated from rats fed ethanol for 7 weeks exhibited decreased retention of calcium in the presence of 4mM-Pi. This defect was associated with enhanced efflux of calcium when mitochondria were incubated with EGTA. Acetaldehyde at low, "physiological" concentrations (100 microM) enhanced calcium retention by mitochondria but this response was blunted after chronic ethanol administration. The in vitro actions of acetaldehyde appear to be mediated, in part, by its metabolism in mitochondria since pretreatment of rats with cyanamide (an aldehyde dehydrogenase inhibitor) prevents this effect.     

Effect of age, vitamin D, and calcium on the regulation of rat intestinal epithelial calcium channels

Transepithelial transport of calcium involves uptake at the apical membrane, movement across the cell, and extrusion at the basolateral membrane. Active vitamin D metabolites regulate the latter two processes by induction of calbindin D and the plasma membrane ATPase (calcium pump), respectively. The expression of calbindin D and the calcium pump declines with age in parallel with transepithelial calcium transport. The apical uptake of calcium is thought to be mediated by the recently cloned calcium channels-CaT1 (or ECaC2, TRPV6) and CaT2 (or ECaC1, TRPV5). The purpose of these studies was to determine whether there were age-related changes in intestinal calcium channel regulation and to identify the dietary factors responsible for their regulation. Young (2 months) and adult (12 months) rats were fed either a high calcium or low calcium diet for 4 weeks. The low calcium diet significantly increased duodenal CaT1 and CaT2 mRNA levels in both age groups, but the levels in the adult were less than half that of the young. The changes in calcium channel expression with age and diet were significantly correlated with duodenal calcium transport and with calbindin D levels. To elucidate the relative roles of serum 1,25(OH)2D3 and calcium in the regulation of calcium channel expression, young rats were fed diets containing varying amounts of calcium and vitamin D. Dietary vitamin D or exogenous 1,25(OH)2D3 more than doubled CaT1 mRNA levels, and this regulation was independent of dietary or serum calcium. These findings suggest that the apical calcium channels, along with calbindin and the calcium pump, may play a role in intestinal calcium transport and its modulation by age, dietary calcium, and 1,25(OH)2D3.     

Tissues and organs as indicators of intestinal absorption of minerals and trace elements,evaluated in rats

Tissue and organ deposition and blood parameters were evaluated as indices of mineral and trace element absorption in rats. The absorption of elements was quantified in relation to nitrogen retention, i.e., considering the weight gain and new tissue synthesis. A rapeseed meal diet was supplied with three levels of calcium, two levels of zinc, and two levels of copper in a factorial design. In general, an increase in dietary mineral content increased the relative absorption, which in turn, increased the tissue deposition progressively. Striated muscle, however, did not respond to either an increased calcium or zinc supply. Furthermore, an increased calcium absorption caused a depression of the fractional phosphorus and magnesium content of femur bones. The copper content of the kidneys and the heart muscle was directly proportional to the amount of absorbed zinc and iron, respectively. The iron content of tissues was, in general, inversely proportional to zinc absorption and showed a tendency to be directly proportional to copper absorption. The zinc level in tissues was, in a similar way, inversely correlated to measured calcium absorption. In conclusion, interactions between elements do not only affect the intestinal element absorption, but also the distribution of already absorbed elements in tissues and organs.     

Temperature-mediated regulation of calcium flux and motility in fowl spermatozoa.

The motility of fowl spermatozoa at various temperatures was shown to be a function of their intracellular calcium content, measured after hypotonic lysis of the cells. Retention of calcium by spermatozoa, with consequent enhancement of motility, increased as the temperature was lowered from 40 degrees to 30 degrees C. Raising the temperature within this range subsequently reduced calcium retention and motility again. The temperature-dependent retention of calcium was a function of the rate of calcium efflux rather then influx. The temperature-sensitive efflux mechanism appeared to involve a Ca2+ ATPase which was relatively inactive at 30 degrees C, but active at 40 degrees C.     

Biological activity assessment of 1 alpha,25-dihydroxyvitamin D3-26,23-lactone in the rat

1 alpha,25-Dihydroxyvitamin D3-26,23-lactone [1 alpha,25(OH)2D3-26,23-lactone] was compared to 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3] in terms of their stimulation, in vivo, of intestinal calcium transport and mobilization of calcium from bone in the rat (the two classic vitamin D-mediated responses), and their relative binding to the chick intestinal receptor for 1 alpha,25(OH)2D3, 1 alpha,25-(OH)2D3-26,23-lactone was found to be only one-thirtieth as active as 1 alpha,25-(OH)2D3 in the stimulation of intestinal calcium transport and was found to mediate a significant reduction in the steady-state serum calcium levels. Associated with the reduction in serum calcium was a significant increase in urinary calcium excretion for 24 h after the administration of the steroid. Prior administration of 1 alpha,25(OH)2D3-26,23-lactone partially blocked the actions of a subsequently administered dose of 1 alpha,25(OH)2D3 in increasing serum calcium levels, but did not affect the action of 1 alpha,25(OH)2D3 in stimulating intestinal calcium transport. The binding affinity of 1 alpha,25(OH)2D3-26,23-lactone to the chick intestinal cytosol receptor protein was observed to be 670 times lower than that of 1,25-(OH)2D3 which indicates that perturbation of the 25-hydroxylated side chain by formation of the 26,23-lactone causes a significant reduction in ligand affinity for the receptor.