Abstract  Recently, [Tc-99m]TRODAT-1, the first Tc-99m-labeled tracer for imaging CNS dopamine transporters in humans, was reported. This tracer displayed excellent specific binding to dopamine transporters in the basal ganglia region of the brain, thus it is potentially useful for the diagnosis of deficit of dopamine transporters in neurodegenerative diseases, such as Parkinson's disease. Preparation of [Tc-99m]TRODAT-1 was previously achieved by a multistep kit formulation. It is highly desirable to further improve the preparation by developing a simplified one-vial formulation with a reduced amount of TRODAT-1 ligand for routine clinical use. To achieve this goal, a series of studies to optimize labeling efficiency by varying a combination of factors (amount of free ligand, reaction reagents, and reaction pH) was carried out. [Tc-99m]TRODAT-1 prepared by this new kit formulation was evaluated by assessing the brain uptake and target (striatum) versus nontarget (cerebellum) ratios in rats. Appropriate amounts of various ingredients for a one-vial kit formulation providing > or =90% radiolabeling yields were identified. The most consistent and reliable formulation contained 10 microg of TRODAT-1 (a reduction of free ligand from 200 microg to 10 microg), 32 microg of SnCl2, 10 mg of sodium glucoheptonate, and 840 microg of disodium EDTA in one vial as a lyophilized kit. It is feasible to reconstitute the vial with [Tc-99m]pertechnetate (0.5-2 mL, < or =1110 MBq, 30 mCi), resulting in a final solution with a pH value of 4.5-5.0. [Tc-99m]TRODAT-1, prepared by this new kit, was stable at room temperature for 6 h. Biodistribution studies of this agent in rats with the new formulation showed similar regional brain distribution as compared with those obtained with the previous preparation (high striatum-to-cerebellum ratio). In conclusion, using this lyophilized one-vial kit formulation, [Tc-99m]TRODAT-1 can be prepared with greater than 90% radiochemical purity. This simplified kit will significantly improve the reliability of preparation of this agent for routine clinical use.

Abstract  Rapid and efficient delivery of radioactive metal complexes to the cell interior would enable novel applications in medical imaging and radiotherapy. Membrane permeant peptide conjugates incorporating HIV-1 Tat transactivation protein sequences (GRKKRRQRRR) and an appropriate peptide-based motif (epsilon-KGC) that provides an N(3)S donor core for chelating technetium and rhenium were synthesized. Oxotechnetium(V) and oxorhenium(V) Tat-peptide complexes were prepared by facile transchelation reactions with permetalates, tin(II) chloride and sodium glucoheptonate. RP-HPLC showed two major [(99m)Tc]Tat-peptide species (4) that differed in retention time by approximately 2 min corresponding to two [Re]Tat-peptide species (7) shown to have identical mass, consistent with formation of two isomers, likely the oxo-metal diastereomers. [(99m)Tc]Tat-peptides were stable to transchelation in vitro. In human Jurkat cells, [(99m)Tc]Tat-peptide 4 showed concentrative cell accumulation (30-fold greater than extracellular concentration) and rapid uptake kinetics (t(1/2) < 2 min) in a diastereomeric-comparable manner. Paradoxically, uptake was enhanced in 4 degrees C buffer compared to 37 degrees C, while depolarization of membrane potential as well as inhibition of microtubule function and vesicular trafficking showed no inhibitory effect. Cells preloaded with 4 showed rapid washout kinetics into peptide-free solution. Modification of [(99m)Tc]Tat-peptide by deletion of the N-terminus Gly with or without biotinylation minimally impacted net cell uptake. In addition, the C-terminus thiol of the prototypic Tat-peptide was labeled with fluorescein-5-maleimide to yield conjugate 8. Fluorescence microscopy directly localized conjugate 8 to the cytosol and nuclei (possibly nucleolus) of human Jurkat, KB 3-1 and KB 8-5 tumor cells. Preliminary imaging studies in mice following intravenous administration of prototypic [(99m)Tc]Tat-peptide 4 showed an initial whole body distribution and rapid clearance by both renal and hepatobiliary excretion. Analysis of murine blood in vivo and human serum ex vivo revealed >95% intact complex, while murine urine in vivo showed 65% parent complex. Thus, these novel Tat-peptide chelate conjugates, capable of forming stable [Tc/Re(V)]complexes, rapidly translocate across cell membranes into intracellular compartments and can be readily derivatized for further targeted applications in molecular imaging and radiotherapy.

Abstract  One novel styrylpyridine derivatives(AV-45) coupled with Tc-99m complex was synthesized. Tc-99m-BAT-AV-45 was prepared by a ligand exchange reaction employing sodium glucoheptonate, and effects of the amount of ligand, stannous chloride, sodium glucoheptonate and pH value of reaction mixture on the radiolabeling yield were studied in details. Quality control was performed by thin layer chromatography and high performance liquid chromatography. Besides the stability, partition coefficient and electrophoresis of Tc-99m-BAT-AV-45 were also investigated. The results showed that the average radiolabeling yield was (95 +/- 1%) and Tc-99m-BAT-AV-45 with suitable lipophilicity was stable and uncharged at physiological pH.

Abstract  The subject of these studies was the thyroid gland tissue of middle-aged (14-month-old) female rats chronically treated with calcium glucoheptonate. The peripheral and central zone of the thyroids were stereologically analysed and the following morphometric parameters determined: the height and volumetric density of follicular epithelium, colloid, interstitium and follicles and index of activation rate. The height of follicular epithelium, its volume density and index of activation rate were significantly reduced (by 8%, p < 0.05, 18%, p < 0.025 and 34%, p < 0.01, respectively) as compared to the controls. However, the volumetric density of colloid and interstitium were increased (by 10% and 14% respectively). These morphometric results indicate that Ca treatment expressed an inhibitory effect on thyroid follicular cells structure in middle-aged female rats.

Abstract  The synthesis and labeling of (99m)Tc-N(3)-{N'-[2-sulfanyl-ethylamino)acetyl]-2-aminoethyl-sulfanyl-1-hexanamide]thymidine ((99m)Tc-NHT) were studied. In the presence of sodium glucoheptonate (GH) and ethylene diamine tetraacetic acid (EDTA), (99m)Tc-NHT was obtained by using bisaminoethanethiol (N(2)S(2)) as a bifunctional coupling agent. The radiochemical purity of the (99m)Tc-NHT was over 95%. Biodistribution of (99m)Tc-NHT was performed in hepatoma HepA tumor-bearing mice. At 2 h p.i., the ratios of tumor-to-muscle, tumor-to-bone and tumor-to-blood were 4.41 +/- 0.32, 2.45 +/- 0.24 and 1.51 +/- 0.18, respectively. (C) 2011 Chun Xiong Lu. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Abstract  The pituitary TSH cell structure of middle-aged (14-month-old) female Wistar rats chronically treated with estradiol dipropionate (EDP), calcium glucoheptonate (Ca) or with the combination of both was studied. TSH-producing cells were examined in the pituitary pars distalis using rabbit anti-rat beta-thyrotropin (TSH) serum and peroxidase-antiperoxidase immunohistochemistry. A stereological method for the determination of morphometric changes of the volume of TSH cells and nuclei as well as of their number and relative volume densities was used. All examined morphometric parameters in treated animals showed a significant decrease in comparison with immunoreactive TSH cells of the controls; the most significant decrease was observed in EDP-treated rats. These results together with structural features of immunoreactive TSH cells in the pituitary of middle-aged rats after chronic application of EDP or Ca indicate that both compounds inhibit these cells.

Abstract  Foliar calcium applications are used in many fruiting crops to minimize disease and physiological disorders. In blueberry (Vaccinium spp), it is used to improve fruit firmness with varying success. Two applications of foliar calcium applied to rabbiteye blueberry (V. virgatum Aiton) cvs. Alapaha and Powderblue as calcium nitrate [Ca(NO3)(2)], neutralized calcium carbonate (CaCO3), and chelated calcium (calcium glucoheptonate, C-14 H-26 CaO16) were made at the label rate of 2.3 L. ha(-1) applied in a volume of 935.3 L. ha(-1) (697 ppm, 108 ppm, and 604 ppm Ca per application, respectively). The applications were made at 30 and 15 days preharvest in 2013 and 2014. Fruit were hand harvested at 40% ripe and evaluated for berry weight, color, firmness, soluble solids, and acidity. In 2013, fruit were stored at 1 degrees C with 85% relative humidity and evaluated again at 7 and 15 days. In 2014, fruit and tissue samples were evaluated for Ca concentration. In 2013, 'Powderblue' had a 5% increase in firmness from the CaCO3 treatment when compared to control fruit. The chelated calcium treatment significantly increased fruit weight by 12% compared to the control for 'Alapaha'. Fruit firmness increased 5% and fruit weight decreased 10% for the Ca(NO3)(2) treatment compared to control for 'Alapaha' fruit sampled after 2 weeks of storage. In 2014, none of the treatments significantly increased fruit firmness or berry calcium concentration. For 'Powderblue' in 2014, all treatments significantly decreased firmness. Leaf Ca concentration was increased by 18% for 'Alapaha' and decreased by 26% for 'Powderblue' when comparing the chelated calcium treatment to non-treated leaves.

Abstract  In the direct labeling of antibodies with Re-186, 188, the lower redox potential of ReO4- than TcO4- requires the addition of excess SnCl2 and a medium-chelating agent for stabilizing the excess of SnCl2 in solution. Through extensive tests, sodium glucoheptonate (GH) was chosen as an excellent stabilizer for SnCl2 and also the reduced Re(V) from a variety of chelators, such as citrate, cyclodextrin, tartrate, inositol, glucose, glycine, etc. ReO4- solution was then quantitatively reduced for 2 h with newly prepared SnCl2(GH) solution. Then, we directly incorporated the reduced Re to the antibodies IgG modified with 135 ford of NaHSO3 and 3500 fold of 2-ME, and more than 90% of specific binding was yielded in 100-150 min at room temperature. TLC analysis indicated that less than 5% of activity was in the colloid form. Radiolabeled antibodies IgG were stable to the challenging of 700 fold of DTPA, and also showed fine in vivo stability.

Abstract  The effects of estradiol dipropionate (EDP) or calcium glucoheptonate (Ca) on the morphology and stereology of the PRL cells in 14-month-old Wistar female rats were studied. The animals were treated daily with EDP in the dose of 0.625 mg/kg b.w. or calcium glucoheptonate (Ca; 11.4 mg/kg b.w) for two weeks. The controls were injected with vehicle alone by the same schedule. Mammotrophs (PRL cells) were immunocytochemically localized by the PAP method. Blood PRL concentration was determined by Delfia procedure. In animals treated with EDP the volume of both, PRL cells and their nuclei, as well as the volume densities were significantly (p<0.05) increased by 17%, 9% and 38%, respectively, in comparison with the controls. In animals treated with Ca all morphometric parameters were insignificantly (p > 0.05) decreased compared to control rats. Serum concentration of PRL was significantly increased (p<0.05) by 17% after estradiol treatment, but in Ca-treated females this parameter was insignificantly (p>0.05) changed by 2% compared to controls. Based on these results, it can be concluded that EDP expresses a strong stimulatory effect on the morphology and function of pituitary PRL cells.

Abstract  The effects of multiple doses of estradiol dipropionate (EDP) or calcium glucoheptonate (Ca) on the growth and function of pituitary somatotropes (GH cells) were studied. Female middle-aged rats were receiving i.p. EDP (0.625 mg i.p./kg b.w), or Ca (11.4 mg/kg b.w) every day for two weeks. Blood samples were collected for hormone analyses and pituitaries dissected for histological and morphometric evaluation 24 h after the last injection. GH-producing cells were examined using the peroxidase-antiperoxidase (PAP) immunohistochemical procedure. Both EDP- and Ca-treatment significantly decreased all morphometric parameters of GH cells (p < 0.05) in comparison with the corresponding controls. Serum concentration of growth hormone (GH) in EDP- or Ca-treated groups was lower by 65% and 13% (p < 0.05) respectively, comparing to the controls. The difference between all morphometric parameters of EDPL and Ca-treated rats was statistically significant (p<0.05) in relation to the controls. These findings suggest that multiple EDP, or Ca application affects (directly or indirectly) the control of growth and secretory activity of GH cells in middle-aged female rats.

Abstract  A precipitate encountered in solutions of calcium gluceptate was identified as hydrated calcium gluceptate. Precipitation was associated with a change from a very soluble amorphous anhydrous form to a sparingly soluble crystalline hydrate, the presence of seed crystals inducing crystallization, and unsuitable proportions of the alpha- and beta-epimers of calcium gluceptate. Various commercial samples and the corresponding precipitates were examined by elemental analysis, thermal analysis, X-ray diffraction, IR spectroscopy, and GC-MS. The proportion of the alpha- and beta-epimers in commercial samples was quantitated by GC. In this method, an aqueous solution of calcium gluceptate was converted into a mixture of glucoheptonic acids and their corresponding lactones by passage through a cation-exchange resin. The solution was freeze-dried, the acid-lactone mixture converted to the gamma-lactones using concentrated hydrochloric acid, and the resulting material trimethylsilylated with trimethylsilylimidazole. Stability studies of solutions prepared from calcium gluceptate obtained from various commercial sources indicate that above approximately 50% alpha-epimer, stability decreased with an increase in the relative proportion of the alpha-epimer. Material complying with USP specifications (pure alpha-epimer) is the least stable in solution. It is suggested that calcium gluceptate containing approximately equal proportions of the alpha- and beta-epimers be introduced in the USP monograph together with a method for estimating the proportions of the epimers.

Abstract  The solubility of sodium naproxen was determined over a range of temperatures from 15.2 degrees C to 39.7 degrees C by two methods: analyses of samples from equilibrated solutions and a recently developed procedure utilizing a focused-beam reflectance method (FBRM). The results demonstrate the utility of the newer and, in some cases, simpler method. A discontinuity in the solubility was observed at 29.8 degrees C, identifying the temperature as which the dihydrate and anhydrous forms of sodium naproxen trade places as being the more stable of the two forms. The heats of solution for the two pseudopolymorphs were obtained from van't Hoff plots of the solubility data. These results were used to demonstrate how the heat of solution of one form can be estimated using the heat of dehydration obtained from differential scanning calorimetry (DSC) and the heat of solution from another form.

Abstract  In the structure of sodium D-glycero-D-gulo-heptonate dihydrate, Na+.C7H13O8-.2H2O, the glucoheptonate anion has a bent carbon chain conformation. There are extensive intermolecular hydrogen bonds involving all the hydroxy and water H atoms. The Na+ cation has a distorted octahedral coordination to six O atoms, with Na+...O distances ranging from 2.316 (2) to 2.645 (2) A.

Abstract  The goal of this study is to develop thermostable microneedle patch formulations for influenza vaccine that can be partially or completely removed from the cold chain. During vaccine drying associated with microneedle patch manufacturing, ammonium acetate and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer salts stabilized influenza vaccine, surfactants had little effect during drying, drying temperature had weak effects on vaccine stability, and drying on polydimethylsiloxane (PDMS) led to increased stability compared with drying on stainless steel. A number of excipients, mostly polysaccharides and some amino acids, further stabilized the influenza vaccine during drying. Over longer time scales of storage, combinations of stabilizers preserved the most vaccine activity. Finally, dissolving microneedle patches formulated with arginine and calcium heptagluconate had no significant activity loss for all three strains of seasonal influenza vaccine during storage at room temperature for 6 months. We conclude that appropriately formulated microneedle patches can exhibit remarkable thermostability that could enable storage and distribution of influenza vaccine outside the cold chain. copyright 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:740-749, 2015

Abstract  The purpose of this study was to investigate the influence of chronic calcium treatment on the structure and function of thyroid C cells in ovariectomzed adult female rats. Eighteen 3-month-old, female Wistar rats were divided into three groups. The first group was used as the sham-operated control, and the other two were surgically ovariectomized (Ovx). One month after gonadectomy, one group of Ovx rats was injected with 28.55 mg Ca-glucoheptonate (Ca)/kg b.w., while the other two groups were chronically treated with vehicle alone (Ovx and sham control). Two months after surgery, the animals were killed. In the thyroid C cells, calcitonin (CT) was localized with the peroxidase-antiperoxidase method. Stereology was used to evaluate morphometric changes in the volume of C cells, their nuclei and relative volume density. The number of C cells per unit area was calculated. Serum CT content was determined by radioimmunoassay. After chronic Ca treatment C cells were numerous with darker cytoplasm than in C cells of sham-operated control animals, but more degranulated than the corresponding cells of Ovx rats. Their volume was significantly decreased by 14% (p < 0.05), while the number was increased by 47% (p < 0.05) in comparison with corresponding controls. Serum CT concentration was decreased by 27% (n.s.) in comparison to sham-operated control. Calcium treatment of Ovx rats led to a 32% increase of serum CT concentration in relation to untreated Ovx animals. These results suggest that chronic Ca treatment of Ovx female rats positively affected CT release from thyroid C cells, without any significant changes in morphometric parameters.

Abstract  The effects of several calcium salts on in vitro growth, pectic enzyme activity, and colonization of excised peach twigs by the peach canker fungus, Leucostoma persoonii, were investigated. Fungal growth was determined on potato-dextrose agar amended with various calcium salts. The greatest growth reduction (85%) was caused by calcium propionate, followed by calcium hydroxide (76%) and calcium silicate (73%). The fungicides captan, iprodione, and thiophanate-methyl completely inhibited fungal growth. Lesion length was reduced when excised peach twigs were wounded with a cork borer, dipped for 15 or 60 min in the various calcium solutions, and inoculated with L. persoonii. For 15-min dips, lesion length was reduced more than 70% by calcium silicate, iprodione, and calcium propionate. After 15 min, calcium in the bark was not greater than that in the distilled, deionized water control. When twig segments were dipped for 60 min, lesion length was reduced more than 70% by calcium acetate, calcium sulfate, calcium heptagluconate, calcium oxide, calcium succinate, calcium chloride, calcium hydroxide, and the fungicide iprodione. No lesions occurred when twigs were treated with calcium propionate, captan, and thiophanate-methyl After 60 min, only twigs dipped in calcium sulfate showed significantly elevated levels of bark Ca2+. Canker length and calcium content of bark were negatively correlated (r = -0.26, P less than or equal to 0.05) after the 15-min treatment. No correlation was found with the 60-min dip, even though calcium content of the bark had increased significantly. When L. persoonii was grown on Ca2+ -amended media, pectin lyase activity was not reduced significantly by any treatment. Calcium oxide and calcium propionate, and calcium silicate treatments reduced polygalacturonase activity after 7 and 15 days, respectively.

Abstract  The effects of multiple treatment with estradiol dipropionate (EDP) or calcium glucoheptonate (Ca) or a combination of the two on gonadotrophic cells in the pituitary pars distalis of middle-aged female rats were examined. The animals were treated daily for two weeks with EDP (0.625 mg i.p./kg body weight) or Ca (11.4 mg/kg body weight) or EDP + Ca. Luteinising (LH) and follicle stimulating hormone (FSH)-producing cells were examined by immunohistochemistry using antisera to the specific (beta) beta-subunits of LH and FSH and a peroxidase-anti-peroxidase immunohistochemical procedure. Plasma levels of FSH and LH were measured by radio-immune assay. A stereological method for determining morphometric parameters in immunopositive FSH and LH cells was used. The number of gonadotrophs per unit area (mm2), their cellular volume and relative volume densities, as well as plasma levels of FSH and LH, were decreased in all treated females in comparison with the controls. The most significant decrease of these parameters was observed in EDP-treated animals. Such changes were also expressed in Ca-treated animals, but the alterations were less distinct. These results demonstrate that multiple EDP or Ca application to middle-aged female rats is able to inhibit, directly or indirectly, the morphofunctional state of gonadotrophic cells in the pituitary pars distalis.