Sol-gel encapsulation of binary Zn(II) compounds in silica nanoparticles. Structure-activity correlations in hybrid materials targeting Zn(II) antibacterial use.

In the emerging issue of enhanced multi-resistant properties in infectious pathogens, new nanomaterials with optimally efficient antibacterial activity and lower toxicity than other species attract considerable research interest. In an effort to develop such efficient antibacterials, we a) synthesized acid-catalyzed silica-gel matrices, b) evaluated the suitability of these matrices as potential carrier materials for controlled release of ZnSO4 and a new Zn(II) binary complex with a suitably designed well-defined Schiff base, and c) investigated structural and textural properties of the nanomaterials. Physicochemical characterization of the (empty-loaded) silica-nanoparticles led to an optimized material configuration linked to the delivery of the encapsulated antibacterial zinc load. Entrapment and drug release studies showed the competence of hybrid nanoparticles with respect to the a) zinc loading capacity, b) congruence with zinc physicochemical attributes, and c) release profile of their zinc load. The material antimicrobial properties were demonstrated against Gram-positive (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus) and negative (Escherichia coli, Pseudomonas aeruginosa, Xanthomonas campestris) bacteria using modified agar diffusion methods. ZnSO4 showed less extensive antimicrobial behavior compared to Zn(II)-Schiff, implying that the Zn(II)-bound ligand enhances zinc antimicrobial properties. All zinc-loaded nanoparticles were less antimicrobially active than zinc compounds alone, as encapsulation controls their release, thereby attenuating their antimicrobial activity. To this end, as the amount of loaded zinc increases, the antimicrobial behavior of the nano-agent improves. Collectively, for the first time, sol-gel zinc-loaded silica-nanoparticles were shown to exhibit well-defined antimicrobial activity, justifying due attention to further development of antibacterial nanotechnology.

An additional middle cuneiform?

Additional cuneiform bones of the foot have been described in reference to the medial bipartite cuneiform or as small accessory ossicles. An additional middle cuneiform has not been previously documented. We present the case of a patient with an additional ossicle that has the appearance and location of an additional middle cuneiform. Recognizing such an anatomical anomaly is essential for ruling out second metatarsal base or middle cuneiform fractures and for the preoperative planning of arthrodesis or open reduction and internal fixation procedures in this anatomical location.

Further investigations of a simple counting technique for measuring mixtures of two pure beta-emitting radionuclides.

A simple liquid scintillation counting technique to measure the activity composition of a mixture containing two known pure beta-emitting radionuclides was recently developed at the NMISA. The method has been applied to various two-component mixtures of (32)P, (33)P and (35)S, primarily to gauge the effect of spectral energy differences on the method's ability to extract the individual activities. Excellent results were obtained for mixtures of (33)P and (35)S, radionuclides with similar, low beta energies. Mixtures containing the high-energy beta-emitter (32)P were more difficult to resolve, although quenching of the counting sources with CHCl(3) improved mixture resolution.

Conformational study of insect adipokinetic hormones using NMR constrained molecular dynamics.

Mem-CC (pGlu-Leu-Asn-Tyr-Ser-Pro-Asp-Trp-NH2), Tem-HrTH (pGlu-Leu-Asn-Phe-Ser-Pro-Asn-Trp-NH2) and Del-CC (pGlu-Leu-Asn-Phe-Ser-Pro-Asn-Trp-Gly-Asn-NH2) are adipokinetic hormones, isolated from the corpora cardiaca of different insect species. These hormones regulate energy metabolism during flight and so are intimately involved in an insect's mobility. Secondary structural elements of these peptides and the N7 analogue, [N7]-Mem-CC (pGlu-Leu-Asn-Tyr-Ser-Pro-Asn-Trp-NH2), have been determined in dimethylsulfoxide solution using NMR restrained molecular mechanic simulations. The neuropeptides were all found to have an extended structure for the first 4 residues and a beta-turn between residues 4-8. For Tem-HrTH and Del-CC, asparagine (N7) which is postulated to be involved in receptor binding and/or activation, projects outward form the beta-turn. Mem-CC does not have an asparagine at position 7 while, for [N7]-Mem-CC, the N7 sidechain folds inside the beta-turn preventing its interaction with the receptor.

Metal-ion speciation in blood plasma incorporating the tetraphosphonate, N,N-dimethylenephosphonate-1-hydroxy-4-aminopropilydenediphosphonate (APDDMP), in therapeutic radiopharmaceuticals.

In a quest for more effective radiopharmaceuticals for pain palliation of metastatic bone cancer, this paper relates results obtained with 166Ho and 153Sm complexed to the bone seeking phosphonate, N,N-dimethylenephosphonate-1-hydroxy-4-aminopropylidenediphosphonate (APDDMP). APDDMP is synthesised from the known bone cancer pain palliation agent 1-hydroxy-3-aminopropylidenediphosphonate (APD) and was complexed to lanthanide trivalent metal ions. This work is performed to utilise the idea that the energetic beta-particle emitter, 166 Ho, coupled with phosphonate ligands such as APD and APDDMP could afford a highly effective radiopharmaceutical in the treatment of bone cancer. Complex-formation constants of APDDMP with the important blood plasma metal-ions, Ca2+, Mg2+, and Zn2+ and the trivalent lanthanides Ho3+ and Sm3+ were measured by glass electrode potentiometry at 37 degrees C and I = 150 mM. Blood plasma models were constructed using the computer code ECCLES and the results compared with those gathered from animal tests. The 166Ho-APDDMP complex was found to have little liver or bone uptake while 153Sm-APDDMP had a moderate bone uptake. This was primarily due to the high affinity of APDDMP for Ca(II). Clinical observations could be explained by the blood plasma modelling.

Design of copper-based anti-inflammatory drugs.

It has been shown that the inflammation associated with rheumatoid arthritis can be reduced using copper complexes. In order to improve the bioavailability of copper and hence efficacy of these complexes we have synthesized three different series of ligands, each having different characteristics. Thermodynamic results for copper(II) complexes for these polyamino, diaminodiamido and triaminodiamido ligands are presented. The polyamino ligands form the most stable complexes in vivo but tissue distribution studies in mice show that [Cu(3,6,9,12-tetraazatetradecanedioate)] is excreted rapidly, unchanged in the urine. The diamino ligand complexes are much less stable than their polyamino analogues and animal studies using [Cu(N,N'-bis[2-(dimethylamino)ethyl]-ethanediamide)H2] indicate that the complex dissociates in vivo and is excreted slowly via the liver. The triaminodiamido copper(II) complexes are approximately 2 log units more stable than their diamino analogues. Computer simulation calculations indicate that these complexes are also likely to dissociate in plasma. Measured partition coefficients, however, suggest the possibility of dermal absorption.

Trace metal excretion in patients with homozygous hypercholesterolaemia.

In patients with familial hypercholesterolaemia regular therapeutic apheresis is acknowledged to have long-term benefit. A previously unrecognised complication of such intervention is the development of anaemia that reflects a sub-optimal dietary iron intake coupled with accelerated loss of this trace metal in the fluid discarded after each procedure. Additional contributions result from enhanced urinary excretion as a result of chelation to citrate used as an anticoagulant and frequent blood sampling. The underlying pathophysiologic process appears to be reduced deformability. We now document similar and significant losses of zinc, copper and chromium in these circumstances. In the case of the latter three elements, no associated clinical syndromes have thus far been identified, probably because deficiency states are less well-recognised than that due to iron loss and, additionally, because critical reductions are avoided by their replenishment during a normal food intake. These studies are, nevertheless, relevant since they are the basis for recommending prophylactic supplementation during this form of management.

Metal-ion speciation in blood plasma incorporating the bisphosphonate, 1-hydroxy-4-aminopropilydenediphosphonate (APD), in therapeutic radiopharmaceuticals.

In the quest for more effective pain palliation radiopharmaceuticals for metastatic bone cancer, this paper relates results obtained with 166Ho complexed to the bone-seeking bisphosphonate, 1-hydroxy-4-aminopropililydenediphosphonate (APD). APD is itself a bone cancer pain palliation agent and this work was therefore driven by the idea that the energetic beta-particle emitter, 166Ho, coupled with APD could afford a highly effective radiopharmaceutical in the treatment of bone cancer. Complex-formation constants for important blood plasma metal-ions were measured by potentiometry or polarography at 37 degrees C and I = 150 mmol dm-3. The latter technique was used for systems where precipitates formed at ligand-to-metal ratios appropriate for potentiometry. For trivalent lanthanides, neither electrochemical technique could be used. Animal tests showed that the 166Ho-APD complex was taken up primarily by the liver due to precipitation or colloid formation.

Metal ion speciation in blood plasma: gallium-67-citrate and MRI contrast agents.

UNLABELLED: Metal chelate ions are commonly used in medical diagnostic imaging as MRI contrast or imaging agents. The efficacy of these metals depends on their in vivo behavior, which in turn depends on their in vivo speciation. METHODS: A computer model has been used to simulate the speciation of Ga3+ and Gd3+ in blood plasma. The model has been tested against known clinical data and then used to investigate Ga3+ uptake by tumor cells. The iatrogenic effect of a gadopentetic acid enhanced MRI scan upon the biodistribution of 67Ga citrate has also been calculated. RESULTS: The speciation of Ga3+ calculated using the computer model is concordant with clinical data. The results support transferrin mediated uptake of Ga3+ by tumor cells but also account for Ga(III) biodistribution observed in hypotransferrinemic subjects. In a study of the effect of gadopentetic acid upon 67Ga gallium citrate, neither residual DTPA nor [Gd(DTPA)]2- cause significant changes in the speciation of Ga(III). The calculations show that dissociation of 4% of the administered gadopentetic acid results in the formation of a mixed, Gd(III) and Ga(III), metal transferrin complex and a 100-fold increase in the concentration of [Ga(OH)4]-. CONCLUSION: Computer simulation is a valuable tool which can be used to explain/understand in vivo behavior of radioactive metal ions.

Hybrid genes over-express pertactin from Bordetella pertussis.

Pertactin is a surface adhesin of Bordetella pertussis which is produced in small quantities when expressed from the native prn promoter. Hybrid genes were constructed in which the prn promoter was replaced by either the fha or tox promoter. Recombinant B. pertussis strains containing chromosomally integrated hybrid tox promoter/prn (toxpprn) or fha promoter/prn (fhapprn) genes expressed pertactin at approximately 5- and 8-fold the wild-type level, respectively. The pertactin was correctly processed and secreted and was biochemically and antigenically comparable to its wild-type counterpart, as determined by N-terminal sequence analysis, immunoblotting, peptide mapping, circular dichroism and antigenicity studies. In an adherence assay, a strain over-expressing pertactin was no more adherent than the wild-type strain, but a pertactin-deficient strain was less adherent.

Formation constants at high ionic strength-I. Potentiometric determination of protonation constants for succinic, propionic and mono-methyl succinic acid in different ionic media.

The protonation constants, K(r), for the ligands succinic acid (SA), mono-methyl succinate (MS) and propionate (PA) have been determined, at 25 degrees C, by glass electrode potentiometry in 3 mol/dm(3) (M) NaNO(3), KNO(3), NH(4)NO(3), Ca(NO(3))(2) and Et(4)NBr aqueous media. Results are compared with literature constants determined in 3M NaClO(4). The order of stability was found to be K(1)(SA) > K(1)(PA) > K(1)(MS) > K(2)(SA) and for the ligands in the different media K(r) followed the general trend with respect to the background electrolyte Et(4)NBr > NaClO(4) > KNO(3) > NaNO(3) > NH(4)NO(3) > Ca(NO(3))(2).

Formation constants at high ionic strength-II. The ionic strength correction of formation constants using a simplified pitzer equation.

A simplified approach to the correction of equilibrium constants from one ionic strength and/or medium to another is developed. A computer program capable of performing these calculations is presented together with results of its application to the NH (3)H (+), CH (3)COO (-)H (+), CH (3)CH (2)COO (-)H (+), - OOCCH (2)COO (-)H (+) and HOOCCH (2)COO (-)H (+) systems.

A prototype recombinant vaccine against respiratory syncytial virus and parainfluenza virus type 3.

We have produced a genetically-engineered chimeric protein composed of the external domains of the respiratory syncytial virus (RSV) fusion (F) protein and the parainfluenza virus type 3 (PIV-3) hemagglutinin-neuraminidase (HN) protein in insect cells using the baculovirus expression system. The yield of the soluble chimeric FRSV-HNPIV-3 protein could be increased approximately 2-fold by using Trichoplasia ni (High Five) insect cells in place of Spodoptera frugiperda (Sf9) for expression. The chimeric protein, purified from the supernatant of baculovirus-infected High Five cells by immunoaffinity chromatography was correctly processed at the F2-F1 proteolytic cleavage site. Immunochemical analysis of the chimera with a panel of anti-F and anti-HN monoclonal antibodies suggested that the antigenicity of the major F and HN neutralization epitopes of the chimeric protein was preserved. Immunization of cotton rats with two 1 or 10 micrograms doses of the chimeric protein adsorbed to aluminum phosphate elicited strong PIV-3 specific HAI responses as well as PIV-3 and RSV specific neutralizing antibodies, and at either dose completely protected against challenge with live RSV and PIV-3.

Technetium-99m labelling of bis-oxime ligands.

The synthesis and labelling of a new bis-amide-oxime ligand E,E-2,9-bis(hydroxyimino)-4,7-diaza 5,6-dioxodecane (AdO) with 99mTc has been achieved. Protein binding, partition coefficient and tissue distribution of this complex and two related bis-amine-oxime ligands is reported. The biodistribution of the complexes are disappointing with only limited brain and myocardial uptake. Structures for the complex are postulated.

Chromium-cage complex as contrast agent in MR imaging--biodistribution studies of the [57Co]cobalt analogue.

The synthesis and T1 and T2 relaxivities of the Cr(III)-(NH2)-Sar-cage complex is reported. An outer-sphere relaxation mechanism is postulated for the relaxivity of the complex. Tissue distribution studies in mice using a [57Co]cobalt analogue as a radioactive tracer showed that the complex is excreted rapidly in the urine. Some renal uptake of the complex is seen. Appreciable uptake of labelled cage complex was observed in 3-methylcholanthrene induced murine rhabdomyosarcoma.

The glycopeptides of the mouse immunoglobulin A T15.

Cleavage of mouse IgA T15 with papain yielded (a) a glycosylated Fab fragment, (b) a non-glycosylated Fc fragment and (c) a glycosylated C-terminal peptide. The cleavage sites at the hinge and at the end of the C alpha 3 domain were located by sequencing. The two glycopeptides were prepared from the Fab and C-terminal fragments by pronase digestion. The C alpha 1 glycopeptide at Asn 155 was complex type with alpha (1-3)galactose terminal groups, and closely resembled the Asn 171 glycopeptide of mouse IgM (Anderson et al. (1985) Arch. Biochem. Biophys. 243, 605-618). In contrast, the C-terminal glycopeptide at Asn 446 was entirely different from the corresponding IgM glycopeptide, being complex rather than high-mannose type.

Gadolinium(III) complex equilibria: the implications for Gd(III) MRI contrast agents.

A computer model of blood plasma which has allowed the effect of Gd(III) contrast agents to be simulated has been developed. Initial binding of Gd(III) is to transferrin. At high concentration the metal ion binds to citrate and salicylate. At a concentrate of 10(-3) M, GdCl3 is predicted to effect a redistribution of the in vivo Zn(II), Ca(II), and Fe(II) complexes present in blood plasma. There is little effect on the Cu(II) distribution. At a concentration below 10(-5) M EDTA and DTPA have little effect on the free Gd(III) metal ion concentration. Above this concentration though, the metal ion is bound almost exclusively to the EDTA or DTPA. An attempt is made to relate the toxicity of GdCl3, [Gd(EDTA)]-, and [Gd(DTPA)]2- to the thermodynamic stability of these complexes. The effect of substitution kinetics is also discussed.

The use of bacteriophage-mediated depolymerisation in investigations of the structure of the capsular polysaccharide from Klebsiella serotype K71.

The structure (1) of the heptasaccharide repeating-unit of the capsular polysaccharide from Klebsiella serotype K71 follows from methylation analysis and n.m.r. and mass-spectrometric studies of the oligosaccharides obtained on depolymerisation of the polysaccharide with a bacteriophage-borne endo-rhamnosidase. [formula; see text].