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Mechanical properties of septal cartilage homografts.

The compressive mechanical properties of untreated and chemically and physically treated nasal septum homografts were determined. Mechanical properties of control, saline-, thimerosal (Merthiolate)- and Alcide-treated specimens were similar. At high strains, the stiffness of treated cartilage ranged from 12.8 to 22.5 MPa and was unaffected by storage time. In comparison, irradiated and freeze-dried nasal septum exhibited stiffnesses of 35 and 37.5 MPa, respectively, after approximately 1 month of storage. These values of stiffness were significantly different from controls at a 0.95 confidence level. On the basis of these results, it was concluded that Alcide and Merthiolate treatment did not alter the compressive mechanical properties of cartilage and that a combination of these treatments may adequately sterilize and preserve nasal septum homografts.

Adolescent↗

Characterization of somatostatin receptors in the rat adrenal glomerulosa zone.

Specific receptors for somatostatin have been identified and characterized in the rat adrenal glomerulosa zone in vivo and in vitro by binding studies with [125I]iodo-Tyr-somatostatin. In adult rats, the injection of [125I]iodo-Tyr-somatostatin was followed by rapid uptake of the labeled peptide in several tissues. The highest uptake was in the adrenal capsule, with a tissue to blood ratio of 4.5, followed by kidney, anterior pituitary, and liver with tissue to blood ratios of 2.0, 1.8, and 1.4, respectively. In vitro binding studies with adrenal capsular particles were performed at 16 C in the presence of bacitracin and thimerosal to reduce tracer degradation. Under these conditions, binding of [125I]iodo-Tyr-somatostatin to capsular membrane-rich fractions reached a steady state within 30-40 min and remained at a plateau for 120 min. Dissociation of bound somatostatin from its adrenal receptors was also rapid, with an initial half-time of 5 min. Equilibrium binding of somatostatin to adrenal capsular particles was saturable and of high affinity, with an association constant (Ka) of 1.5 x 10(10) M-1. The receptor-binding activities of several somatostatin analogs were consistent with their potencies as inhibitors of angiotensin II-stimulated aldosterone production in adrenal capsular cells and with their known biological activities upon GH release. These findings demonstrate that high affinity receptors with structural and biological specificities for somatostatin are present in the adrenal glomerulosa zone. Such receptors serve as the regulatory sites through which somatostatin inhibits the action of angiotensin II upon aldosterone production in the adrenal glomerulosa cell.

Adrenal Glands↗

Analgesic effects of intra-nasal enkephalins.

The analgesic effects of intranasal delivery of leucine enkephalin (Leu-Enk) and its synthetic analogue [D-ala(2)]-leucine enkephalinamide (YAGFL) with or without enzyme inhibitors and/or absorption enhancers were investigated using the acetic acid-induced writhing test in mice. The analgesic activity was significantly affected by the time delay after the administration of Leu-Enk; the inhibition rates for the groups administered with acetic acid 5 min and 30 min after the administration of Leu-Enk were 56.40 +/- 8.54 and 17.98 +/- 7.07%, respectively. The addition of enzyme inhibitors and absorption enhancers markedly increased the inhibition rate of Leu-Enk and YAGFL; their inhibition rates were about four times and twice those without any enzyme inhibitor or absorption enhancer, respectively. The enzyme inhibitors and absorption enhancers that produced the highest inhibition rates of Leu-Enk and YAGFL were azelaic acid (1%), thimerosal (0.5 mM, TM), ethylenediamine-tetraacetic acid (5 mM, EDTA) and L-alpha-lysophosphatidylcholine (0.5%, LPC), and TM (0.5 mM), EDTA (5 mM), LPC (0.5%) and povidone (5%), respectively. The ED50 value of both enkephalins was also determined and found to be about 13 microg kg(-1), which is 850 and 60 times more potent than literature values for ketoprofen and morphine, respectively. Based on these results it was concluded that Leu-Enk or YAGFL could exert very high analgesic activity when administered nasally with a combination of inhibitors and absorption enhancers as compared with other analgesics.

Acetic Acid↗

Thiomersal enhances the binding of histamine to the H1 receptor, but not histamine-stimulated inositol phosphate formation.

Thiomersal (thimerosal) was a weak inhibitor of the binding of [(3)H]mepyramine to histamine H(1) receptors in guinea-pig cerebellar membranes (11 +/- 1% inhibition at 10 microM, 32 +/- 3% inhibition at 300 microM). However, in the concentration range 3-30 microM, thiomersal enhanced the binding of histamine to the H(1) receptor, as reflected by the displacement of curves of histamine inhibition of [(3)H]mepyramine binding to lower concentrations, without any change in the Hill coefficient. The ratio of the IC50 values (the concentration giving 50% inhibition) in the absence and presence of thiomersal increased from 1.8 with 3 microM to 3.6 with 30 microM thiomersal. There was no consistent effect of thiomersal at concentrations of 30 microM and below on curves of mepyramine inhibition of [(3)H]mepyramine binding. In the presence of 10 microM thiomersal histamine-induced accumulation of inositol phosphates in U373 MG astrocytoma cells was partially inhibited (37 +/- 8% inhibition of the maximum response), without any significant change in the EC50 (the concentration giving the half maximal response) for histamine. Thus although histamine binding was potentiated by thiomersal, there was no potentiation of an H(1) receptor-mediated functional response.

Animals↗

A direct measurement of increased divalent cation influx in fertilised mouse oocytes.

On fertilisation of mouse oocytes, the fusing spermatozoon triggers a series of repetitive calcium (Ca2+) spikes. The Ca2+ spikes seem to be necessary for successful progression through the cell cycle and are regulated in a cell-cycle-dependent manner. The spikes appear to require the linkage of continuous Ca2+ influx to the periodic release of Ca2+ from intracellular stores by a process of Ca(2+)-induced Ca2+ release. The precise role of Ca2+ influx was explored using the manganese (Mn2+)-quench technique to monitor unidirectional cation influx into single mouse oocytes. There was a marked stimulation of cation influx associated closely with the upsweep of the first and subsequent fertilisation Ca2+ spikes. A smaller but significant increase in the rate of cation influx persisted in the interspike period in fertilised oocytes. Spike-associated entry was not as apparent in oocytes stimulated to spike repetitively by thimerosal or acetylcholine application. Instead, there was a continuous increase in cation influx underlying Ca2+ spiking which commenced with the onset of the first spike. Using the specific microsomal inhibitor thapsigargin and the Ca2+ ionophore ionomycin, we found evidence for a capacitative entry mechanism in mouse oocytes. We propose that the persistent influx of Ca2+ observed in response to all stimuli examined is controlled by a capacitative mechanism and sets the frequency of spiking by determining the time taken to refill the internal stores to a point where they are again sensitive enough to initiate the next spike.

Acetylcholine↗

The three modern faces of mercury.

The three modern "faces" of mercury are our perceptions of risk from the exposure of billions of people to methyl mercury in fish, mercury vapor from amalgam tooth fillings, and ethyl mercury in the form of thimerosal added as an antiseptic to widely used vaccines. In this article I review human exposure to and the toxicology of each of these three species of mercury. Mechanisms of action are discussed where possible. Key gaps in our current knowledge are identified from the points of view both of risk assessment and of mechanisms of action.

Animals↗

When science is not enough - a risk/benefit profile of thiomersal-containing vaccines.

Without a preservative, such as thiomersal (known as thimerosal in the US), multi-dose liquid presentations of vaccine are vulnerable to bacteriological contamination that can result in death or serious illness of the recipient. Concerns about levels of mercury exposure from thiomersal-containing vaccines were first raised in the US during 1999 in the context of Hepatitis B vaccine for newborns. Since then, a large body of evidence from animal and epidemiological studies has accumulated on the safety of thiomersal. Ironically, these data have become largely irrelevant in wealthy countries, where mono-dose, thiomersal-free vaccines have been introduced as a precautionary measure in almost all childhood vaccines, in part related to residual public scepticism. In poor countries, multi-dose vials remain important for vaccine delivery. There is a real danger that this controversy may result in the loss to the world of thiomersal as a preservative, simply from popular pressure. In reality, it would be impossible to cease overnight using thiomersal and maintain the supply of vital vaccines. This paper reviews and summarises the data available from published studies on mercury toxicity, and thiomersal in vaccines in particular, that overwhelmingly indicate continued use of thiomersal is safe in those countries where it is most needed.

Adverse Drug Reaction Reporting Systems↗

Postlicensure epidemiology of childhood vaccination: the Danish experience.

The efficacy, the ability to confer protection against a target disease and the safety of a vaccine are assessed in great detail before licensure. However, inherent limitations in the prelicensure assessment necessitate continued epidemiological evaluations of efficacy and safety issues after the introduction of vaccines into use. In Denmark, the opportunities available for epidemiological research are unique. In 2001, an initiative was undertaken to take advantage of these opportunities to study the postlicensure epidemiology of childhood vaccination with respect to effectiveness and safety. First, we describe the unique opportunities for postlicensure research in Denmark with respect to the data sources available and the epidemiological and statistical methods used. We then describe a number of recent postlicensure studies of effectiveness and safety that took advantage of these opportunities. Specifically, studies on the effectiveness of Haemophilus influenzae type b vaccination, the effectiveness of pertussis vaccination, the impact of a preschool pertussis booster on infant pertussis, measles-mumps-rubella vaccine and autism, thimerosal-containing vaccine and autism, measles-mumps-rubella vaccine and febrile seizures, childhood vaccination and Type 1 diabetes, and childhood vaccination and nontargeted infectious disease are discussed.

Autistic Disorder↗

Cell-mediated cytotoxicity-supporting activity of various human gammaglobulin preparations.

Antibody activity, especially that involved in the reaction of antibody-dependent cell-mediated cytotoxicity (ADCC), of five commercially available human gammaglobulin preparations (standard, pepsin-treated, plasmin-treated, polyethylene glycol-fractionated and S-sulfonated gammaglobulin) was measured. All these gammaglobulin preparations had high titers of hemagglutination inhibition and neutralizing antibody against measles virus. In ADCC reaction, the pepsin-treated gammaglobulin preparation showed no antibody activity. The standard gammaglobulin preparation showed weak activity only when highly diluted. The remaining three preparations showed high activity. Though the S-sulfonated gammaglobulin preparation showed no activity in ADCC reaction, it showed high activity after reconversion by means of oxidation and reduction in vitro. The plasmin-treated gammaglobulin preparation showed greater activity than the polyethylene glycol-fractionated preparation of the optimal concentration. In ADCC tests using the plasmin-treated gammaglobulin preparation, K cell activity was strongly inhibited by Hg (thimerosal), while, in those using the standard gammaglobulin preparation, the activity was hardly influenced by Hg, suggesting that the low ADCC activity of the standard gammaglobulin preparation of high concentrations was due to the inhibitory effect of aggregated immunoglobulin G molecules.

Antibody-Dependent Cell Cytotoxicity↗

Thiomersal in vaccines: balancing the risk of adverse effects with the risk of vaccine-preventable disease.

A number of affluent countries are moving to eliminate thiomersal (thimerosal), an ethylmercury preservative, from vaccines as a precautionary measure because of concerns about the potential adverse effects of mercury in infants. The WHO advocates continued use of thiomersal-containing vaccines in developing countries because of their effectiveness, safety, low cost, wide availability and logistical suitability in this setting. The guidelines for long-term mercury exposure should not be used for evaluating risk from intermittent single day exposures, such as immunisation using thiomersal-containing vaccines. Similar or higher mercury exposures likely occur from breast feeding and the health benefit of eliminating thiomersal from a vaccine, if any, is likely to be very small. On the other hand, the benefits accrued from the use of thiomersal-containing vaccines are considerably greater but vary substantially between affluent and developing regions of the world. Because of the contribution to overall mercury exposure from breast milk and diet in later life, the removal of thiomersal from vaccines would produce no more than a 50% reduction of mercury exposure in infancy and <1% reduction over a lifetime. Different public policy decisions are appropriate in different settings to achieve the lowest net risk, viewed from the perspectives of the individual vaccinee or on a population basis. In developing regions of the world, at least over the next decade, far more benefit will accrue from protecting children against widely prevalent vaccine-preventable diseases by focusing efforts aimed at improving infant immunisation uptake by using current, inexpensive, domestically-manufactured, thiomersal-containing vaccines, than by investing in thiomersal-free alternatives.

Animals↗

[Photosensitivity induced by piroxicam].

In Portugal, piroxicam is one of the most prescribed non-steroidal anti-inflammatory agents (NSAIDs) and is frequently associated to photosensitivity skin reactions, well known by all dermatologists. The authors review the physiopathology and clinical patterns of this adverse drug reaction. Importance is given to the high prevalence of thimerosal sensitisation in our country, a marker of piroxicam photosensitivity. The authors propose that piroxicam prescription should be avoided in Portugal, as this drug can be substituted by any other NSAID, without marked photosensitivity.

Anti-Inflammatory Agents, Non-Steroidal↗

Methods used to evaluate the effectiveness of contact lens care solutions and other compounds against Acanthamoeba: a review of the literature.

PURPOSE: The purpose of this paper is to review the literature concerning the methods used to evaluate contact lens care solutions against Acanthamoeba. Acanthamoeba keratitis is a potential threat, with 85% of the cases being reported in contact lens wearers. METHODS: Several studies from the published literature that evaluated contact lens disinfectants were reviewed. The variables included test organism, strain and morphology, growth conditions, inoculum preparation, inoculation method, test solutions and concentration, contact time, neutralization, recovery, quantitation method, and viability determination of survivors. The methods used to test Acanthamoeba against the disinfectants were compared and contrasted. RESULTS: After a thorough review of methods used to test Acanthamoeba, it was found that there is great variability in the methods used to evaluate contact lens disinfectants. The majority of the studies used A.castellanii and A.polyphaga cysts grown axenically in PYG medium containing cations at about 30 degrees C and the inoculum contained about 1.0 x 10(5) cells/mL. Inactivation media or centrifugation of cells was used to neutralize test samples. Quantitation was performed in most studies and viability was checked in all studies. The disinfectants tested most often were PHMB, hydrogen peroxide, thimerosal, and chlorhexidine. CONCLUSIONS: After reviewing the studies presented here it can be concluded that an effective method for testing Acanthamoeba against contact lens disinfectants would include A.castellanii or A.polyphaga grown axenically in PYG containing cations and a concentration of organisms high enough to adequately measure kill, a neutralization step, recovery and quantitation of organisms followed by a viability check of survivors.

Acanthamoeba↗

Reduction of the Ca2+ transport systems in mouse young brown fat cells.

Principal differences in the kinetics and amplitude of Ca2+ response to norepinephrine were found between freshly isolated young differentiated brown fat cells. An increase in the Ca2+ concentration in the cytoplasm ([Ca2+]i) in the young cells was unusually slow (A[Ca2+]i = 0.03 nM/s) in comparison with that in the differentiated cells, and the Ca2+ influx from the outside was not induced by Ca2+ mobilization agents, such as thapsigargin and ionomycin. Ionomycin increased [Ca2+]i up to 150 nM in a Ca2+-free medium and up to 270 nM in the normal medium. This results in that the intracellular Ca2+ stores in freshly isolated young cells are rather poor, and the mechanism of capacitive calcium entry does not virtually function. The data on chemical modification of Ca2+ channels in the plasma membrane by thimerosal suggest that the conductance of these channels is low and/or their number in young brown fat cells is insignificant.

Adipocytes↗

[Involvement of protein kinase C in the regulation of Ca2+ exit from intracellular stores of prolactin stimulated pig oocytes].

Involvement of protein kinase C in the regulation of Ca2+ exit from intracellular stores of pig oocytes activated by prolactin was investigated, using the fluorescent dye chlortetracycline. In the presence of extracellular calcium, the inhibitor of protein kinase C Ro 31-8220 increased calcium exit from intracellular stores in pig oocytes after prolactin treatment. In calcium-free medium, Ro 31-8220 exerted effect on calcium release from intracellular stores. In calcium-free medium, prolactin did not stimulate calcium release from intracellular stores of oocytes in the presence of thimerosal, while in the presence of protein kinase C inhibitor, prolactin increased Ca2+ content from intracellular stores in such oocytes. These data suggest a direct involvement of protein kinase C in the processes of regulation of Ca2+ exit from intracellular stores of pig oocytes stimulated by prolactin.

Animals↗

Vaccine adverse events: separating myth from reality.

Vaccines have turned many childhood diseases into distant memories in industrialized countries. However, questions have been raised about the safety of some vaccines because of rare but serious adverse effects that have been attributed to them. Pain, swelling, and redness at the injection site are common local reactions to vaccines. Fever and irritability may occur after some immunizations. Currently, no substantial evidence links measles-mumps-rubella vaccine to autism, or hepatitis B vaccine to multiple sclerosis. Thimerosal is being eliminated from routine childhood vaccines because of concerns that multiple immunizations with vaccines containing this preservative could exceed recommended mercury exposures. Family physicians should be knowledgeable about vaccines so that they can inform their patients of the benefits of immunization and any proven risks. If immunization rates fall, the incidence of vaccine-preventable illnesses may rise.

Adverse Drug Reaction Reporting Systems↗

Efficacy studies on Haemophilus gallinarum bacterin preparations.

Broth cultures inactivated and potentiated by selected methods were tested in chickens for efficacy against homologous and heterologous challenge inoculation, using 2 serotype A strains of Haemophilus gallinarum. Although the 2 strains were within the same serotype, they failed to cross protect. One dose of thimerosal-inactivated bacterin was protective against homologous challenge, but 2 doses of formalin-inactivated bacterin were required. A bivalent bacterin protected chickens well against 1 strain, but not the other, at the 1-dose level.

Animals↗

[Inhibition effect of merthiolate used as vaccines preservative on the oxygen-dependent metabolism of neutrophil granulocytes].

The influence of the newly developed complex vaccine Pyopol, containing the antigens of opportunistic microorganisms and polyoxydonium used as immunomodulator, on the oxygen-dependent metabolism of neutrophils was studied. The study revealed that the main components of the vaccine, both individually and in association, did not change cellular activity in the range of concentrations used in this study. The inhibition of the oxygen-dependent metabolism of neutrophil granulocytes in the presence of native or weakly diluted vaccine occurred due to the cytotoxic effect of thimerosal used as preservative.

Adjuvants, Immunologic↗

[Mercury in vaccines].

Thiomersal, also called thimerosal, is an ethyl mercury derivative used as a preservative to prevent bacterial contamination of multidose vaccine vials after they have been opened. Exposure to low doses of thiomersal has essentially been associated with hypersensitivity reactions. Nevertheless there is no evidence that allergy to thiomersal could be induced by thiomersal-containing vaccines. Allergy to thiomersal is usually of delayed-hypersensitivity type, but its detection through cutaneous tests is not very reliable. Hypersensitivity to thiomersal is not considered as a contraindication to the use of thiomersal-containing vaccines. In 1999 in the USA, thiomersal was present in approximately 30 different childhood vaccines, whereas there were only 2 in France. Although there were no evidence of neurological toxicity in infants related to the use of thiomersal-containing vaccines, the FDA considered that the cumulative dose of mercury received by young infants following vaccination was high enough (although lower than the FDA threshold for methyl mercury) to request vaccine manufacturers to remove thiomersal from vaccine formulations. Since 2002, all childhood vaccines used in Europe and the USA are thiomersal-free or contain only minute amounts of thiomersal. Recently published studies have shown that the mercury levels in the blood, faeces and urine of children who had received thiomersal-containing vaccines were much lower than those accepted by the American Environmental Protection Agency. It has also been demonstrated that the elimination of mercury in children was much faster than what was expected on the basis of studies conducted with methyl mercury originating from food. Recently, the hypothesis that mercury contained in vaccines could be the cause of autism and other neurological developmental disorders created a new debate in the medical community and the general public. To date, none of the epidemiological studies conducted in Europe and elsewhere support this assumption. Although any effort should be made to avoid useless exposure of vaccinees to a potentially toxic compound, it should be emphasized that 1) public communication on this issue has led to a decrease in the hepatitis B vaccination coverage of children born to HBs Ag positive mothers in the US; 2) this issue was not really relevant in France where until 2002, apart from two hepatitis B vaccines, all childhood vaccines were thiomersal-free, and 3) in developing countries using multidose vaccine vials, moving to thiomersal-free vaccines in unidose presentations would represent such an incremental cost that millions of children would no more have access to vaccination. Therefore the World Health Organisation still recommends the use of thiomersal-containing vaccines as part of the expanded programme of immunisation.

Autistic Disorder↗