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Regulation of rubber biosynthetic rate and molecular weight in Hevea brasiliensis by metal cofactor.

Metal ion cofactors are necessary for prenyltransferase enzymes. Magnesium and manganese can be used as metal ion cofactor by rubber transferase (a cis-prenyltransferase) associated with purified rubber particles. The rubber initiation rate, biosynthetic rate, and molecular weight produced in vitro from Hevea brasiliensis rubber transferase is regulated by metal ion concentration. In addition, varies significantly with [Mg(2+)]. decreases from 8000 +/- 600 microM at [Mg(2+)] = 4 mM to 68 +/- 10 microM at [Mg(2+)] = 8 mM and increases back to 970 +/- 70 microM at [Mg(2+)] = 30 mM. The highest affinity of rubber transferase for IPP.Mg occurred when [Mg(2+)] = A(max) (metal concentration that gives highest IPP incorporation rate). A metal ion is required for rubber biosynthesis, but an excess of metal ions interacts with the rubber transferase inhibiting its activity. The results suggest that H. brasiliensis could use [Mg(2+)] as a regulatory mechanism for rubber biosynthesis and molecular weight in vivo.

Hemiterpenes↗

Biotechnological possibilities for waste tyre-rubber treatment.

Every year large amounts of spent rubber material, mainly from rubber tyres, are discarded. Of the annual total global production of rubber material, which amounts to 16-17 million tonnes, approximately 65% is used for the production of tyres. About 250 millions spent car tyres are generated yearly in USA only. This huge amount of waste rubber material is an environmental problem of great concern. Various ways to remediate the problem have been proposed. Among these are road fillings and combustion in kilns. Spent tyres, however, comprise valuable material that could be recycled if a proper technique can be developed. One way of recycling old tyres is to blend ground spent rubber with virgin material followed by vulcanization. The main obstacle to this recycling is bad adhesion between the crumb and matrix of virgin rubber material due to little formation of interfacial sulphur crosslinks. Micro-organisms able to break sulphur-sulphur and sulphur-carbon bonds can be used to devulcanize waste rubber in order to make polymer chains on the surface more flexible and facilitate increased binding upon vulcanization. Several species belonging to both Bacteria and Archaea have this ability. Mainly sulphur oxidizing species, such as different species of the genus Thiobacillus and thermoacidophiles of the order of Sulfolobales, have been studied in this context. The present paper will give a background to the problem and an overview of the biotechnological possibilities for solutions of waste rubber as an environmental problem, focusing on microbial desulphurization.

Bacteria↗

Molecular cloning, expression and characterization of cDNA encoding cis-prenyltransferases from Hevea brasiliensis. A key factor participating in natural rubber biosynthesis.

Natural rubber from Hevea brasiliensis is a high molecular mass polymer of isoprene units with cis-configuration. The enzyme responsible for the cis-1,4-polymerization of isoprene units has been idengified as a particle-bound rubber transferase, but no gene encoding this enzyme has been cloned from rubber-producing plants. By using sequence information from the conserved regions of cis-prenyl chain elongating enzymes that were cloned recently, we have isolated and characterized cDNAs from H. brasiliensis for a functional factor participating in natural rubber biosynthesis. Sequence analysis revealed that all of the five highly conserved regions among cis-prenyl chain elongating enzymes were found in the protein sequences of the Hevea cis-prenyltransferase. Northern blot analysis indicated that the transcript(s) of the Hevea cis-prenyltransferase were expressed predominantly in the latex as compared with other Hevea tissues examined. In vitro rubber transferase assays using the recombinant gene product overexpressed in Escherichia coli revealed that the enzyme catalyzed the formation of long chain polyprenyl products with approximate sizes of 2 x 103-1 x 104 Da. Moreover, in the presence of washed bottom fraction particles from latex, the rubber transferase activity producing rubber product of high molecular size was increased. These results suggest that the Hevea cis-prenyltransferase might require certain activation factors in the washed bottom fraction particles for the production of high molecular mass rubber.

Amino Acid Sequence↗

Mutagenicity of rubber vulcanization gases in Salmonella typhimurium.

Gases formed by rubber and rubber additives in the vulcanization process were collected with a laboratory-scale glass apparatus. Mutagenicity testing of the vulcanization gases by the Salmonella/microsome test was conducted with strains TA1535, TA1538, TA98, and TA100 in the absence and presence of a metabolizing system from rat liver homogenates. The mutagenicity of gases derived by heating chloroprene rubber and ethylene propylene rubber was established with both base substitution- and frameshift-sensitive strains and that of a styrene-butadiene rubber was established with the base substitution-sensitive stain TA100. Tests on pyrolysis gases from a butadiene acrylonitrile rubber revealed only toxic effects. Curing systems, additives, and filling materials from various sources were represented in the material. Gases were collected at temperature levels corresponding to both mixing and curing of these particular rubbers in the industrial operations. Attempts were made to correlate the mutagenicity of the gases to the presence of mutagenic components in the rubber mixtures.

Acrylonitrile↗

Glutaraldehyde: its uptake by sporing and non-sporing bacteria, rubber, plastic and an endoscope.

Uptake of glutaraldehyde to bacterial spores, germinating and outgrowing spores, vegetative cells (sporing and non-sporing bacteria), various types of rubber, plastic and an endoscope was investigated. Escherichia coli NCTC 10418 exhibited greatest uptake, followed by Bacillus subtilis NCTC 8236 vegetative cells and Staphylococcus aureus NCTC 6571. Germinated and outgrowing B. subtilis spores adsorbed more glutaraldehyde than resting spores, but less than vegetative cells. Low concentrations of alkaline and acid glutaraldehyde increased the surface hydrophobicity and inhibited the germination of bacterial spores, the alkaline solution to a greater extent in both cases. Rubbers exhibited varying degrees of uptake and are listed in decreasing order of uptake: red rubber, fluorinated rubber (Vinescol), silicone rubber (Silescol), butyl rubber (Butyl XX). Polypropylene, the only plastic examined, was found not to adsorb any glutaraldehyde. The endoscope adsorbed more glutaraldehyde (per gram) than fluorinated rubber but less than red rubber. No damage was observed.

Aldehydes↗

Occurrence of coring in insulin vials and possibility of rubber piece contamination by self-injection.

Coring is reported to occur because rubber pieces are shaved off from a rubber stopper when a needle is inserted into the rubber stopper of transfusion liquid formulation. We verified whether coring really occurs in insulin vials of self-injecting patients. We collected insulin cartridges from 30 hospitalized patients and used the primary injection (trial injection), the secondary injection and the cartridge remaining preparation as samples. We observed the rubber pieces using a microscope and measured the shape, number of pieces. The occurrence rate of coring was 73% for the primary injection, 47% for the secondary injection and 97% for the cartridge remaining preparation. The rubber pieces in the primary injection and the secondary injection which went through the needle are mostly in aggregate shape and the rubber pieces in the cartridge remaining preparation which did not go through the needle are mostly in needle-like shape. A number of small rubber pieces are found in both the primary injection and the secondary injection, indicating a high possibility that rubber pieces may be injected under subcutaneous tissue. The coring is considered to occur because needles are repeatedly inserted and rotated at the same spot. It is required to improve the structure to mount a needle to the pen-type injector in future. Coring is a very serious problem from the medical and pharmaceutical points of view. Further study should be made on the implication to latex allergy and lipodystrophy.

Adult↗

Influence of environmental stress on the kinetics and strength of attachment of Listeria monocytogenes Scott A to Buna-N rubber and stainless steel.

Attachment and detachment of Listeria monocytogenes Scott A to Buna-N rubber and stainless steel under varying conditions of temperature and pH were investigated using model systems. Numbers of attached cells increased with increasing attachment temperature (10 to 45 degrees C) and time (up to 120 min) for both test surfaces. Compared to Buna-N rubber, the rate of attachment to stainless steel was markedly more rapid for all temperature and pH conditions studied and could not be calculated. Rate of attachment to Buna-N rubber was found to be significantly lower when cells were attached at 10 degrees C. Growth temperature did not significantly affect rates of adhesion to Buna-N rubber. Altering the medium pH during attachment between 4 and 9 demonstrated that rates of adhesion were slower under alkaline conditions. Growth pH was also found to significantly affect rates of attachment to Buna-N rubber. Detachment of cells adhered to Buna-N rubber was significantly affected by growth temperature but not growth pH. Significant differences in detachment were also found between Buna-N rubber and stainless steel, inferring stronger attachment to Buna-N rubber. Cell surface hydrophobicity was found to be affected by both growth temperature and growth pH. However, changes in hydrophobicity could not be correlated to differences in rates of attachment. Addition of 0.01% trypsin to the attachment medium during cell exposure to either test surface resulted in a 99.9% reduction in the adhered cell population when compared to controls. This would suggest that proteins play a role in the initial attachment process of L. monocytogenes.

Bacterial Adhesion↗

Use of rubber dam and its association with other endodontic procedures in New Zealand.

The use of rubber dam for endodontic treatment in New Zealand was analysed by a national survey of general dental practitioners. The response rate was 79 percent. Rubber dam was used routinely by 57 percent, its use increasing among practitioners graduating since 1969. A greater percentage of practitioners with less than 10 years experience used rubber dam than did more experienced practitioners. Use of rubber dam was associated with sodium hypochlorite and EDTA as canal irrigants. Reamers were the favoured hand instrument of non-users of rubber dam. No significant differences were found in the canal obturation techniques of users and non-users, but rubber dam users used the long-cone paralleling method of radiography significantly more than non-users. Practitioners in solo practice used rubber dam significantly less than those in group practice. Rubber dam users attended significantly more refresher courses in endodontics than non-users.

Chi-Square Distribution↗

133Xenon absorption into rubber-protected portable cadmium telluride (CdTe(Cl)) detectors invalidating the 133Xenon washout method for measurement of cutaneous and subcutaneous blood flow rates in man.

The importance of 133Xenon absorption into rubber detector caps during cutaneous and subcutaneous blood flow measurement was investigated in 46 experiments involving 38 persons. 133Xenon was administered atraumatically. Cutaneous and subcutaneous washout rates were registered by portable Cadmium Telluride detectors without rubber caps, with rubber caps, and with rubber caps with Mylar membranes interposed between the rubber and the tissue investigated. No difference in rate constants obtained by means of various detector types was detected. The accumulation of 133Xenon in the rubber caps was found to take place within the first few minutes after the detectors had been brought into position. The 133Xenon then diffused back into the tissue exhibiting a great variation regarding rate constants. The 133Xenon diffused form rubber into air and perfused tissue tracing a monoexponential course; and again the rate constants would vary considerably. No correlation was found between elimination rates obtained with detectors with and without 133Xenon polluted caps, and no way of correcting for the 133Xenon content in the rubber caps was found. Relative changes in rate constants could still be recognized, but absolute values were not obtainable.

Absorption↗

[Hygienic evaluation of new brands of rubber intended for use in a water-supply system under high temperature and active chemical operating modes].

The paper presents the results of hygienic testing of new rubbers, IRP-2043 and IRP-1136. The rubber IRP-2043 does not change the quality of the water it contacts with. Water extracts of this rubber do not produce a toxic effect on homoiothermal animals. This rubber can therefore be recommended to be used in potable water supply systems. The rubber IRP-1136, when contacting with dechlorinated tap water, alters its organoleptic and physico-chemical properties. This rubber cannot be recommended for water supply systems. After a prolonged (18 months) exposure to the condensate of degradation products of the agent 0-30, urine conserving agent and a mixture of urine and the conserving agent the rubbers IRP-2043 and IRP-1136 do not alter physico-chemical and bacteriological properties of water or disturb physiological functions of homoiothermal animals. Having this in view, both rubbers can be used in water supply systems functioning at high temperatures and under the influence of active chemicals.

Animals↗

Nitrosamines and rubber.

Occupational exposure to N-nitrosamines in the rubber industry was first reported by Fajen et al. (1979). In order to study the origin and formation of nitrosamines in this industry, chemicals and industrial products, as well as the air in various working areas, were analysed (Spiegelhalder et al., 1980). All chemicals used for rubber compounding contain nitrosamines if they are derivatives of secondary amines; e.g., tetramethylthiurame, zinc-diethyldithiocarbamate or N-oxydiethylene benzothiazolylsulfenamide. All rubber products containing these dialkyl amine derivatives exhibited considerable levels of the corresponding nitrosamines. Accordingly, variable concentrations of airborne nitrosamines could be detected at places where rubber products are manufactured or stored. The nitrosamines found correspond to the compounded chemicals. The original nitrosamine level in rubber chemicals is not high enough to explain the amounts found in rubber products and in air, so that additional nitrosation must occur. The responsible nitrosating agents are described. Preliminary results show that, in most cases, the elimination of nitrosating agents or the use of different rubber chemicals can drastically reduce nitrosamine levels in rubber products and in working areas.

Air Pollutants↗

[Respiratory function in rubber-processing workers].

The prevalence of acute and chronic respiratory symptoms and diseases as well as lung function changes were studied in 409 rubber male workers and 172 nonexposed control male workers. Ventilatory capacity was measured by recording maximum expiratory flow-volume (MEFV) curves on which forced vital capacity (FVC), one second forced expiratory volume (FEV1) and flow rates at 50% and the last 25% of vital capacity were calculated (FEF50, FEF25). A significantly higher prevalence of all chronic respiratory symptoms except asthma was found in rubber workers in comparison to the controls. Smokers had significantly higher prevalence of most of the chronic respiratory symptoms than nonsmokers. Rubber workers exposed for more than 10 years had also significantly higher prevalence of chronic respiratory symptoms than those with shorter exposure. A high prevalence of acute symptoms during work shift was recorded in rubber workers. Ventilatory capacity was mostly decreased during work shift, particularly FEF50 and FEF25. All acute reductions were greater in rubber workers exposed for more than 10 years as compared to those with shorter exposure. Alupent inhaled at the end of the work shift significantly improved decreased values. Rubber workers demonstrated significantly lower ventilatory capacity tests with respect to the controls. A large number of rubber workers showed a decrease in lung functions tests less than 70% of the predicted normal values. Our data suggest that rubber workers are exposed to numerous noxious agents which may lead to acute and/or chronic impairment of lung function.

Adolescent↗

Silicone rubber tubulization in peripheral sensory nerve reconstruction: an experimental study in rabbits.

Silicone rubber (polydimethyl siloxane) tubes are used clinically in peripheral nerve reconstruction. A disadvantage of this procedure is that the material often has to be removed owing to its mechanical properties. The aim of our study was to investigate the healing of reconstructed sensory nerves tubulized by silicone rubber in an animal model. In our experiments, we reconstructed the saphenous nerves in 27 rabbits. In series 1 (n = 12), silicone rubber tubes were slid over a nerve suture without a gap. In series 2 (n = 12), silicone rubber tubes were slid over a 10-mm nerve gap. In series 3 (n = 12), conventional suturing was performed in the collateral saphenous nerves of the animals of the series 1. Epineurial suturing was performed. Three other collateral nonoperated saphenous nerves served as controls. The healing was studied after 3, 6, and 12 months. Morphometric analysis of the regenerating axons was performed by using our new method for quantification of nerve fibers in cross sections stained by immunohistochemistry and using confocal laser scanning microscopy. Data analysis was carried out using a software program especially developed for this purpose. Our results showed in the silicone procedures that at 12 months significantly fewer axons per fascicle area were present compared with conventional suturing. However, mean axon diameters in the distal nerve stump of the silicone procedures did not differ significantly compared with the conventional suturing procedure. The ratio of total axon area to total fascicle area in the distal nerve stumps of the silicone procedure without gap was significantly smaller compared with the conventionally sutured nerve. The percentage outgrowing axons from the proximal nerve stump into the distal one in the silicone rubber procedure without gap was 57%. This was significantly higher than in the silicone rubber procedure with 10-mm gap (48%). However, in conventional suturing, the percentage of outgrowing axons (99%) was significantly higher than in both tubulization procedures. It appeared that tubulization by silicone rubber of sutured nerves without gap did not enhance axon regeneration. Conventional suturing gave significantly better results. If a gap was present, the use of a silicone rubber tube was preferable to non-suturing.

Animals↗

Surface characterization of recycled tire rubber to be used in cement paste matrix.

The surface modification of tire rubber after treatment with saturated NaOH aqueous solution was investigated by HATR infrared analysis, potentiometric titration, and contact angle measurements. Infrared analysis of the powdered treated rubber showed a decrease in absorption at 1540, 1450, and 1395 cm(-1). This decrease is attributed to the removal of zinc stearate, an additive present in tire formulations that often migrates and diffuses to the surface, resulting in poor adhesion between the rubber and other materials. The potentiometric titration of the suspension of powdered rubber in 0.1 M NaCl showed that more hydrochloric acid was consumed by the untreated rubber, most likely a result of the hyrdrolysis of the zinc stearate to the organic acid. Contact angles of flat tire pieces showed an homogeneity enhancement of the treated rubber surface. The decrease of the zinc stearate on the treated rubber surface explains the improvement in the adhesion of this material to the cement matrix, observed in a previous research. The promising results of this study are a starting point for future research on incorporating rubber particles into cementitious materials as a means of successfully utilizing the vast amounts of tire waste currently in landfills.

Journal Article↗

Triple rubber band ligation for hemorrhoids: prospective, randomized trial of use of local anesthetic injection.

PURPOSE: Rubber band ligation is a common office procedure for hemorrhoids. Triple rubber band ligation in a single session has been shown to be a safe and economical way of treating hemorrhoids. However, postligation discomfort after triple rubber band ligation is not uncommon. The aim of this study was to evaluate the effectiveness of local anesthetic injection to the banded hemorrhoidal tissue in reducing postligation discomfort. METHODS: Patients attending an outpatient clinic for symptomatic hemorrhoids suitable for triple rubber band ligation were randomly assigned to two groups. In the treatment group rubber band ligation was performed at three columns of hemorrhoids, and 1 to 2 ml of 2 percent lignocaine was injected into the banded hemorrhoidal tissue. In the control group triple rubber band ligation was performed in a similar manner, but local anesthetic was not given. Patients were followed up by telephone at the second week and in the clinic after six weeks. RESULTS: From April to August 1996, 101 patients entered the trial and were treated with triple rubber band ligation. Sixty-two patients were randomly assigned to the local anesthetic injection group and 39 to the control group. Overall good to excellent results occurred in 89 percent of patients, and there was no difference between the two groups. Postligation pain occurred in 26 and 20 percent of patients in the treatment and control groups, respectively (P > 0.05). Postligation tenesmus occurred in 32 and 41 percent of patients in the treatment and control groups, respectively (P > 0.05). No patients suffered from septic complications or bleeding that required transfusion. CONCLUSION: Triple rubber band ligation in a single session is a safe, economical, and effective way of treating symptomatic hemorrhoids. Postligation pain and tenesmus occurred in 24 and 37 percent, respectively. Discomfort was usually tolerable. Local anesthetic injection to the banded hemorrhoidal tissue did not help to reduce postligation discomfort.

Anesthetics, Local↗

Randomized trial of rubber band ligation vs. stapled hemorrhoidectomy for prolapsed piles.

PURPOSE: The introduction of stapled hemorrhoidectomy may replace local techniques such as rubber band ligation as a first-line treatment for Grade III and small Grade IV piles. We conducted a randomized trial to determine the role of rubber band ligation in the era of stapled hemorrhoidectomy. METHODS: Fifty-five patients with Grade III or small Grade IV hemorrhoids were randomly allocated to either rubber band ligation or stapled hemorrhoidectomy. Patient demographics and procedure-related details were recorded. Follow-up was at two weeks and two and six months to assess complications, symptom relief, incontinence scores, quality of life, and patient satisfaction. RESULTS: Twenty-five patients were randomly assigned to rubber band ligation and 30 to stapled hemorrhoidectomy. The groups were equally matched for age, gender, grade of piles, continence scores, and quality of life. Stapled hemorrhoidectomy was associated with increased pain and analgesia usage at both 2-week and 2-month follow-up (P < 0.001). Rubber band ligation and stapled hemorrhoidectomy were equally effective in controlling symptomatic prolapse, but rubber band ligation was associated with an increased incidence of recurrent bleeding (P = 0.002). There were 6 procedure-related complications in the stapled hemorrhoidectomy group compared with none in the rubber band ligation group (P = 0.027). There was no difference between the two groups in terms of continence scores, patient satisfaction, or quality of life. CONCLUSION: Stapled hemorrhoidectomy is associated with more pain and minor morbidity than rubber band ligation in the treatment of Grade III and small Grade IV piles. However, for those patients who do not want the risk of further intervention procedures, stapled hemorrhoidectomy offers the better chance of a symptomatic cure.

Adult↗

Efficacy of tray adhesives for the adhesion of elastomer rubber impression materials to impression modeling plastics for border molding.

STATEMENT OF PROBLEM: Tray adhesive, which is used for the adhesion of elastomer rubber impression materials to a custom resin tray, lowers the retention of the impression materials to the impression modeling plastics, as some ingredients of tray adhesive make the impression modeling plastic soft and tacky. PURPOSE: The efficacy of tray adhesive, which is used for the adhesion of elastomer rubber impression materials to a custom resin tray, on the adhesion between elastomer rubber impression material and impression modeling plastic was investigated. MATERIAL AND METHODS: Four silicone rubber impression materials (two addition reaction types and two condensation reaction types), two polysulfide rubber impression materials, and one impression modeling plastic were used in this study. Tensile strength between elastomer rubber impression material and impression modeling plastic with or without the application of tray adhesive was evaluated. RESULTS: Although tray adhesives for both addition reaction type and both condensation reaction type of silicone impression materials and one tray adhesive for polysulfide rubber impression material increased the tensile strength between the impression material and impression modeling plastic, one tray adhesive for polysulfide rubber impression material decreased the tensile strength when sufficient drying time was not applied.

Adhesiveness↗

Protein farnesyltransferase inhibitors interfere with farnesyl diphosphate binding by rubber transferase.

Rubber transferase, a cis-prenyltransferase, catalyzes the addition of thousands of isopentenyl diphosphate (IPP) molecules to an allylic diphosphate initiator, such as farnesyl diphosphate (FPP, 1), in the presence of a divalent metal cofactor. In an effort to characterize the catalytic site of rubber transferase, the effects of two types of protein farnesyltransferase inhibitors, several chaetomellic acid A analogs (2, 4-7) and alpha-hydroxyfarnesylphosphonic acid (3), on the ability of rubber transferase to add IPP to the allylic diphosphate initiator were determined. Both types of compounds inhibited the activity of rubber transferases from Hevea brasiliensis and Parthenium argentatum, but there were species-specific differences in the inhibition of rubber transferases by these compounds. Several shorter analogs of chaetomellic acid A did not inhibit rubber transferase activity, even though the analogs contained chemical features that are present in an elongating rubber molecule. These results indicate that the initiator-binding site in rubber transferase shares similar features to FPP binding sites in other enzymes.

Alkyl and Aryl Transferases↗