Protein diffusion patterns in human saliva

BACKGROUND: There are two modes of saliva protein diffusion on a cellulose flat matrix. The monophasic mode of diffusion consists in a homogeneous distribution of salivary protein components on the matrix. On the contrary, in the biphasic mode, an area occupied by nondiffusible proteins is surrounded by an area occupied by diffusible proteins. AIM: To study protein diffusion patterns of saliva obtained from normal human volunteers. MATERIAL AND METHODS: Saliva was obtained from 33 subjects aged 67.5 +/- 12 years old (29 female). Forty microliters of saliva were deposited in the center of cellulose disks for protein diffusion assays. RESULTS: Thirty three whole saliva samples and 31 submandibular/sublingual samples showed a biphasic diffusion pattern. On the contrary, 62 out of 66 samples of parotid saliva displayed a monophasic diffusion pattern. In one parotid saliva samples and 2 submandibular/sublingual samples, the diffusion pattern could not be established. Patterns of salivary protein diffusion were highly consistent within the same individual. Eighty percent of subjects had an unequivocal pattern of saliva protein diffusion. CONCLUSIONS: The monophasic mode of saliva protein diffusion is a feature of parotid saliva, whereas the biphasic mode is characteristic of submandibular/sublingual and whole saliva.     

Detection of anti-HIV-1 IgG antibodies in whole saliva by GACELISA and Western blot assays

The present study, based on 158 HIV seropositives and 167 HIV seronegatives, demonstrates that saliva collected with the Omni-SAL device and tested with GACELISA (an IgG antibody capture ELISA) is an effective non-invasive alternative to serum for anti-HIV IgG antibody screening. The study also shows that a conventional serum Western blot kit can be used, with slight modifications, for confirmatory testing of saliva specimens. Collecting saliva with the Omni-SAL device had a very good acceptance rate among Tanzanian subjects, and although this diagnostic method is not yet known by the general public, 65% of the study participants preferred to give saliva instead of blood for HIV testing.     

Frequent isolation of human herpesvirus 7 from saliva

BACKGROUND: The purpose of this study was to investigate the influence of a prophylactic bone protective treatment with the bisphosphonate dichloromethane/diphosphonic acid (Cl2MBP) in an experimental model of osteolysis with intraosseous implantation of the Walker carcinosarcoma 256 B. METHODS: The biphosphonate was given as a high-dose, short-term treatment (30 mg/5 days sc) followed by a treatment-free interval (1-7 weeks) or as a low-dose, long-term prophylactic treatment (2.5 or 5.0 mg/day/3 weeks sc). Osteolysis was measured with a radiographic and histologic grading system. RESULTS: The high-dose short-term prophylactic treatment was shown clearly to inhibit tumor osteolysis. The osteoprotective effect decreases with increasing length of the treatment-free interval. A similar positive result could be achieved following the low-dose long-term prophylactic treatment. Dosage could not be shown to influence the inhibition of tumor osteolysis in the long-term bone protective treatment. A possible direct influence of the treatment on tumor growth could be ruled out. The prophylactic treatment does not inhibit body weight increase. Animals treated prophylactically showed less weight loss than the controls after tumor implantation. CONCLUSIONS: These results show that a prophylactic treatment with Cl2MBPs protects the skeleton effectively against tumor osteolysis.     

Simultaneous determination of catechins in human saliva by high-performance liquid chromatography

Green tea extracts have been suggested to possess a preventive effect against dental caries. A quantitative method for their anticariogenic substances, catechins, was developed to evaluate their concentrations in human saliva after mouthrinsing with green tea extract. Salivary catechins were extracted to the organic phase after forming a complex with diphenylborate and an ion-pair with tetra-n-butylammonium, and then back-extracted to the acidic aqueous phase. The extract was analyzed by high-performance liquid chromatography using diode array detection at absorption wavelengths ranging from 269 to 278 nm. In reversed-phase chromatography by a gradient elution, eight catechins originating from green tea and an internal standard were separated in 15 min without interfering peaks. All the catechins were simultaneously and selectively determined in the concentration range 0.05-25.0 microg/ml. In replicate spiking experiments with standards, the mean recovery ranged between 86 and 99%, and both intra- and inter-assay C.V.s were within 2.3%. When mouthrinsing with an aqueous solution of green tea extract (5.0 mg/ml) containing eight catechins, the quantitative results revealed that each catechin was retained at microg/ml levels in saliva for up to 60 min.     

Gustin from human parotid saliva is carbonic anhydrase VI

It has been found that yeast mutants deficient in cytosolic superoxide dismutase CuZnSOD are hypersensitive to ferrous iron. In contrast mutants that are deficient in catalases and cytochrome c peroxidase do not differ from the standard strain in this respect. These findings suggest that iron toxicity may depend on the redox status of the cell. They also shed light on the role of superoxide dismutases in preventing the toxic effects of oxygen.     

Wounds of the hand contaminated by human or animal saliva

A prospective and retrospective evaluation of 75 patients with hand wounds contaminated by human saliva (35) or animal saliva (40) demonstrates that a program of outpatient management can be sufficient for optimal care in many patients. This series challenges the proposition that hospitalization, radiographs, and surgical debridement are necessary for most such wounds. Sixty-seven per cent did not have surgical intervention and no complications resulted. Ninety-two per cent received antibiotics. Radiographs were obtained only when bony injury or entry into a joint was suspected. Delay in seeking treatment until obvious signs of infection or pain are present is common. Literature review details the anatomic factors important in the natural history and control of these infections, and the changes with respect to modes of treatment for these potentially dangerous wounds. The injury is caused by bites with the hand extended or, in fight-bite wounds, with the metacarpal-phalangeal and interphalangeal joints flexed, allowing deeper penetration and then sealing of the wound when the first is opened. Staphylococcus and Streptococcus are the organisms most frequently found in human bites, and in animal bites; Pasteurella multocida should be considered in dog and cat bites.     

Relation between zinc content in saliva and blood in healthy human adults

Measurements were made of zinc levels in saliva, whole blood and plasma as well as copper levels in plasma by atomic absorption spectrophotometry. The mean zinc concentration (in ppm) was 0.478 (n = 136) in resting mixed saliva, 0.046 (n = 36) in parotid saliva, 6.41 (n = 145) in whole blood and 0.87 (n = 145) in plasma. The whole blood zinc levels were significantly lower in females than in males (P less than 0.001). Females and blood donors (males) had a significantly (P less than 0.001) higher Cu:Zn ratio than males (non-donors). It would thus appear, first, that the local effect of salivary zinc cannot be assessed simply from the concentration of zinc in stimulated parotid saliva since about nine-tenths of the zinc in resting mixed saliva is derived from other sources and, second, that the Cu:Zn in plasma is the most eligible variable for detecting sub-clinical Zn deficiency.     

Origin and mechanisms of non-disjunction in human autosomal trisomies

The selective functions of D3 receptors in the brain are still poorly understood, mainly because all the ligands active at dopamine D3 receptors have also a high affinity for the D2 receptors. However, it is possible to study selectively D3 receptor function because some brain structures, such as the islands of Calleja, contain D3 and not D2 receptors. The position of the island of Calleja Magna in the rat brain makes it possible to inject dopamine D3 ligands into the vicinity of these D3 receptors, and to study their behavioral role, with no concomitant action on D2 receptors. We studied the effects on body temperature and on locomotion of unilateral microinjections of D2/D3 receptors ligands into the island of Calleja Magna and into the adjacent nucleus accumbens. The results show that D3 agonists injected into the island of Calleja Magna decrease body temperature and that this effect is potentiated by simultaneous injection of the D1 agonist SKF 38393. D3 agonists have no effect on locomotor activity in the island of Calleja Magna. In the nucleus accumbens, the D3 agonists have only weak effects on body temperature, but, when associated with a D1 agonist, strongly stimulate locomotor activity. The effects on body temperature of unilateral microinjections of dopamine agonists into unilaterally dopamine-depleted animals are the same as those in nondepleted ones. This indicates that the D3 receptors are localized postsynaptically in the island of Calleja Magna.     

Effects of hyperthyroidism and radioactive iodine given to ablate the thyroid on the composition of whole stimulated saliva

In recent years, a growing interest in the study of peptide antigenicity in relation to the role of flanking sequences and protein topology in processing, presentation, and recognition has been observed. However, the information available on the antigenicity of recombinant fusion proteins and their effect on the selection of antigen receptor repertoires is limited. To analyze the role of molecular topology of T epitopes in a system relevant to human pathology, we have used the bacterially expressed Schistosoma japonicum glutathione S transferase (GST) to construct recombinant antigens containing HIV-1 derived T cell determinants, and human T cell clones specific for these determinants. We found that antigenicity of a given GST-peptide combination was not the same when T cells and antigen presenting cells from different individuals were tested. Our results show that differences in processing and presentation of chimeric proteins are not dictated by the use of diverse restriction elements. We also found that the context in which an antigenic peptide is delivered affects the recruited repertoire as defined according to T cell receptor V beta usage and fine specificities of selected T cells.     

Peroxynitrite augments fMLP-stimulated chemiluminescence by neutrophils in human whole blood

The neutrophil respiratory burst was examined by the technique of luminol-dependent chemiluminescence (LDCL) triggered by submaximal concentrations of N-formyl-methionyl-leucyl-phenylalanine (fMLP) in diluted whole blood. We sought to identify the chemical species responsible for LDCL in whole blood, to examine the role of leukotriene B4 (LTB4) and other arachidonic acid metabolites as mediators of the fMLP signaling pathway, and to investigate the effect of peroxynitrite on this response. Both sodium azide and taurine significantly inhibited LDCL (93% inhibition with 100 microM azide, 52% inhibition with 10 mM taurine). More modest inhibition was seen with superoxide dismutase (SOD), catalase, the nitric oxide synthase inhibitor monomethyl-L-arginine (L-NMMA), and with inhibitors of the cyclooxygenase (indomethacin), lipoxygenase (AA-861; no effect), and cytochrome P-450 (SKF 525-A) pathways of arachidonic acid metabolism. The nitric oxide donor SIN-1 (1-100 microM) and peroxynitrite (10-300 microM) also augmented fMLP-induced LDCL. The augmentation seen with peroxynitrite and SIN-1 was attenuated by SOD. Despite the increase in LDCL, peroxynitrite caused a dose-related inhibition of fMLP-stimulated LTB4 release. In summary, our results indicate that (1) LDCL elicited by fMLP in diluted whole blood appears primarily mediated by hypochlorous acid derived from myeloperoxidase; (2) pretreatment with the nitric oxide donor SIN-1 or with peroxynitrite augments LDCL; and (3) LTB4 release does not contribute to fMLP-stimulated LDCL or in the modulation of LDCL by SIN-1 or peroxynitrite.     

Processing of adenosine receptor agonists in rat and human whole blood

A stability study of adenosine receptor agonists in rat and human whole blood was performed. The compounds were incubated at 37 degrees in fresh blood, and aliquots of the incubation mixture were hemolyzed at regular time intervals and analyzed with HPLC. N6-cyclopentyladenosine (CPA) and N6-cyclobutyladenosine (CBA) were degraded, whereas N6-cyclohexyladenosine, N6-cycloheptyladenosine and N6-sulfophenyladenosine were not. 2-Chloroadenosine had a half-life very similar to that of CPA. However, the 2'-, 3'-, and 5'-deoxyribose derivatives of CPA remained intact. The nucleoside transport inhibitor nitrobenzylthioinosine attenuated CBA and CPA metabolism in rat blood as did the inhibitor of adenosine deaminase erythro-9-(2-hydroxy-3-nonyl)adenine, albeit at relatively high concentrations. Complete blockade of CBA and CPA degradation was achieved by a preincubation of rat and human blood with the adenosine kinase (AK) inhibitor 5'-amino-5'-deoxyadenosine. We conclude that the two adenosine analogues are metabolized by AK both in rat and in human whole blood.     

Lipoxygenase products in human saliva: patients with oral cancer compared to controls

Porcine colipase, the protein cofactor of pancreatic lipase, was isolated from pancreas freshly collected on animals and from a side fraction from the production of insulin (Novo Nordisk A/S). Samples of purified colipase were analyzed for homogeneity by polyacrylamide gel electrophoresis, reverse-phase high-performance liquid chromatography (RPLC), quantitative N-terminal sequence determination and mass spectrometry. The activating properties of colipase preparations were assayed against tributyrin, triolein or the commercial Intralipid emulsion, in presence of bile salt. Two fractions of colipase with the same specific activity were purified from fresh pancreas. The major fraction (85%) contained one single protein corresponding to fragment 1-93 of the 95-residue form of colipase (procolipase) previously characterized in porcine pancreatic juice. The other fraction (15%) corresponded to fragment 1-91 of procolipase. Also, two fractions of colipase were purified from the side fraction supplied by Novo. These fractions consisted of the 95-residue proform of colipase and of fragment 1-93, respectively, both specifically cleaved at the Ile79-Thr80 peptide bond with partial removal of isoleucine at position 79 and serine at position 78. Procolipase split at the 79-80 bond retained full activity on tributyrin and triolein and on the Intralipid emulsion but the kinetics of hydrolysis of triacylglycerol substrates showed much longer lag periods than those observed with native procolipase. Also, all forms of procolipase split at the 79-80 bond showed one peak in RPLC but their retention time was markedly decreased as compared to that of native procolipase which indicated a weaker hydrophobic binding capacity. The value of the retention time was of the same order of magnitude as that of inactive reduced procolipase. Treatment of native procolipase by pancreatic endopeptidases showed that elastase is likely responsible for specific cleavage at the 79-80 bond of procolipase purified from the Novo extract. Limited proteolysis by trypsin of the proforms of colipase split at the 79-80 bond reduced the lag period. Results presented in this communication provide the first direct evidence showing that the finger-shaped peptide segment between half-cystine residues at positions 69 and 87 is involved in colipase-lipid interaction as previously hypothesized from the three-dimensional structure of the protein.     

Properties of whole saliva and dental plaque in relation to 40-month consumption of chewing gums containing xylitol, sorbitol of sucrose

The concern with nursing research in the future work of nurse graduates. Expectations from the viewpoint of the nurse researcher. Keeping in mind the concerns which occupy nurse researchers now and in the future, expectations are raised of the graduates of degree programmes in nursing science which are now also being established in the Federal Republic of Germany. This takes into account the structurally determined areas of self determined action in new fields of professional practice. Nurse graduates will be seen as "change agents" in their future areas of work and will have three essential aspects of involvement with nursing research: 1. they themselves will conduct research 2. they will receive, put into practice and transmit results of nursing research 3. they will initiate and commission nursing research projects. According to the broad topics of nursing research, requirements for nurses to act as professionals in their new working fields are formulated referring to the given structural conditions in Germany.     

Effect of macrolides on cytokine mRNA expression in human whole blood model

Recently, low dose and long term use of Macrolides (Mls) has been reported to be effective in treatment of chronic lower respiratory tract infections, however its mechanism is still obscure. We evaluated the effect of Mls (EM, AZM, RKM) on cytokine mRNA expressions. We preincubated the whole blood with several concentrations of Mls and removed the Mls and then stimulated human whole blood with LPS as an experimental vivo model. In order to examine cytokine mRNA expressions, we used the RT-PCR method. Cytokine mRNA expressions were suppressed significantly (p < 0.05) by pretreatment with EM, AZM; moreover, the suppression was peaked at low concentrations (0.04 approximately 0.2 microgram/ml). Although, Cytokine mRNA expressions were not suppressed by pretreatment with RKM. These results suggest that EM, AZM have suppression on Cytokine mRNA expressions, and consequently, this suppression has a reasonable effect for DPB patients.     

Interleukin-18 enhances lipopolysaccharide-induced interferon-gamma production in human whole blood cultures

Interleukin-18 (IL-18) is a newly described cytokine, formerly called interferon-gamma (IFN-gamma)-inducing factor. In a simple 24-h human whole blood culture, IFN-gamma was produced by the combination of lipopolysaccharide (LPS) plus IL-18. To liberate cytokines in the leukocyte and red cell compartments, the detergent Triton X-100 was added to the entire blood culture. The combination of low concentrations of LPS plus IL-18 induced a 3- to 5-fold greater production of IFN-gamma than did either stimulant alone. Tumor necrosis factor-alpha (TNF-alpha), IL-6, and IL-8 were also produced. The presence of IL-10 completely suppressed the production of IFN-gamma and reduced that of TNF-alpha, IL-6, and IL-8. Thus, IFN-gamma, TNF-alpha, IL-8, and IL-6 are produced in a single whole blood culture, making correlations in the synthesis of a T helper type 1 cytokine and proinflammatory cytokines with disease activity possible in a single culture.     

Control of whole body balance in the frontal plane during human walking

A whole-body inverted pendulum model was used to investigate the control of balance and posture in the frontal plane during human walking. The model assessed the effects of net joint moments, joint accelerations and gravitational forces acting about the supporting foot and hip. Three video cameras and two force platforms were used to collect kinematic and kinetic data from repeat trials on four subjects during natural walking. An inverse solution was used to calculate net joint moments and powers. Whole body balance was ensured by the centre of mass (CM) passing medial to the supporting foot, thus creating a continual state of dynamic imbalance towards the centerline of the plane of progression. The medial acceleration of the CM was primarily generated by a gravitational moment about the supporting foot, whose magnitude was established at initial contact by the lateral placement of the new supporting foot relative to the horizontal location of the CM. Balance of the trunk and swing leg about the supporting hip was maintained by an active hip abduction moment, which recognized the contribution of the passive accelerational moment, and countered a large destabilizing gravitational moment. Posture of the upper trunk was regulated by the spinal lateral flexors. Interactions between the supporting foot and hip musculature to permit variability in strategies used to maintain balance were identified. Possible control strategies and muscle activation synergies are discussed.