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PubMed2006

Initiator-independent and initiator-dependent rubber biosynthesis in Ficus elastica.

The rubber-producing tree, Ficus elastica (the Indian rubber tree), requires the same substrates for rubber production as other rubber-producing plants, such as Hevea brasiliensis (the Brazilian or Para rubber tree), the major source of commercial natural rubber in the world, and Parthenium argentatum (guayule), a widely studied alternative for natural rubber production currently under commercial development. Rubber biosynthesis can be studied, in vitro, using purified, enzymatically active rubber particles, an initiator such as FPP, IPP as the source of monomer, and a metal cofactor such as Mg2+. However, unlike H. brasiliensis and P. argentatum, we show that enzymatically active rubber particles purified from F. elastica are able to synthesize rubber, in vitro, in the absence of added initiator. In this paper, we characterize, for the first time, the kinetic differences between initiator-dependent rubber biosynthesis, and initiator-independent rubber biosynthesis, and the effect of cofactor concentration on both of these processes.

Ficus↗
PubMed2004

A novel cDNA from Parthenium argentatum Gray enhances the rubber biosynthetic activity in vitro.

Natural rubber (cis-1,4-polyisoprene) is an isoprenoid compound produced exclusively in plants by the action of rubber transferase. Despite a keen interest in revealing the mechanisms of rubber chain elongation and chain length determination, the molecular nature of rubber transferase has not yet been identified. A recent report has revealed that a 24 kDa protein tightly associated with the small rubber particles of Hevea brasiliensis, therefore designated small rubber particle protein (SRPP), plays a positive role in rubber biosynthesis. Since guayule (Parthenium argentatum Gray) produces natural rubber similar in size to H. brasiliensis, it is of critical interest to investigate whether guayule contains a similar protein to the SRPP. A cDNA clone has been isolated in guayule that shares a sequence homology with the SRPP, thus designated guayule homologue of SRPP (GHS), and the catalytic function of the protein was characterized. Sequence analysis revealed that the GHS is highly homologous in several conserved regions to the SRPP (50% identity). In vitro functional analysis of the recombinant protein overexpressed in E. coli revealed that the GHS plays a positive role in isopentenyl diphosphate incorporation into high molecular weight rubbers as SRPP does. These results indicate that guayule and Hevea rubber trees contain a protein that is similar in its amino acid sequence and plays a role in isopentenyl diphosphate incorporation in vitro, implying that it contributes to the enhancement of rubber biosynthetic activity in rubber trees.

Allergens↗
PubMed1997

Results of patch testing with a special series of rubber allergens.

The purpose of this study was to examine the results of patch testing with the rubber components on a standard screening tray and compare them with the results of testing with a special series of 27 rubber components (rubber tray). 1670 patients were patch tested with the screening tray and 317 of these were also tested with the rubber tray. 16% of those tested with the rubber tray had a positive response to at least 1 of the rubber allergens on the screening tray and 22% had a positive response to at least 1 of the allergens on the rubber tray. The most common positive response to the rubber tray allergens was to tetramethylthiuram monosulfide. There were no responses to 3 of the components on the rubber tray and there was only 1 positive response to a further 4 components. The diagnostic test characteristics of the rubber components on the screening tray were examined using the rubber tray as the gold standard. The sensitivity of the screening tray was 94%, specificity 51%, positive predictive value 87% and negative predictive value 71%. Of the 317 tested, 11% were found to have a positive to a substance on the rubber tray that was not evident from the results of the screening tray.

Adult↗
PubMed1989

Purification of a prenyltransferase that elongates cis-polyisoprene rubber from the latex of Hevea brasiliensis.

We have purified "rubber transferase" from latex of the commercial rubber tree Hevea brasiliensis and find that it is a dimer with a monomeric molecular mass of 38,000 Da, requires Mg2+, and is stabilized by thiols in agreement with studies of a partially purified preparation previously described (Archer, B. L., and Cockbain, E. G. (1969) Methods Enzymol. 15, 476-480). Greater than 90% of the [1-14C]isopentenyl pyrophosphate which is incorporated into deproteinated rubber particles by the purified prenyltransferase is added to high molecular mass polyisoprene (greater than 20,000 Da). Purified prenyltransferase and deproteinated rubber particles reconstitute 40-60% of the biosynthetic activity of whole latex in samples matched for rubber content. Incorporation is linear with added rubber particles up to at least 10 mg/ml rubber or 20 microM rubber molecules (based on a number average molecular mass of 500,000 Da). Prenyltransferase concentrations estimated in whole latex (0.37% or 160 nM) are sufficient to saturate all elongation sites in whole latex, and addition of purified prenyltransferase does not increase [1-14C]isopentenyl pyrophosphate incorporation. Deproteinated rubber particles can be titrated with the pure enzyme (Kd = 9 nM) demonstrating that the fraction of rubber molecules available for addition is low (approximately 0.01%). An estimated 7,000 isoprene units are added per complex at a rate of 1/s in a typical assay. Hevea prenyltransferase catalyzes the formation of cis-isoprene in the presence of rubber particles. However, in the absence of rubber particles and in the presence of dimethylallyl pyrophosphate, the purified prenyltransferase catalyzes the formation of geranyl pyrophosphate and all trans-farnesyl pyrophosphate as demonstrated by thin layer chromatography, gas chromatography, and molecular exclusion chromatography.

Amino Acid Sequence↗
PubMed1988

Occupational vitiligo due to unsuspected presence of phenolic antioxidant byproducts in commercial bulk rubber.

We investigated the occurrence of cutaneous depigmentation (vitiligo) among employees of a company that manufactured hydraulic pumps. The interiors of these pumps were injection-molded with rubber. We identified a small but significant cluster of vitiligo cases among a group of employees who frequently handled the rubber used in this injection molding process. Although none of the additives specified in the rubber formulations was a phenolic or catecholic derivative, known to be potential causes of chemically induced vitiligo, gas chromatographic analysis identified a para-substituted phenol (2,4-di-tert-butylphenol, DTBP) in solid samples of the most frequently used rubber. Surface wipe analysis confirmed that workers could be exposed to DTBP from simple handling of the rubber. We subsequently established that the solid bulk rubber used as the base in these stock rubber formulations contained both DTBP and smaller quantities of p-tert-butylphenol. Both had formed as unsuspected byproducts during chemical synthesis of two antioxidants added to the solid bulk rubber by a major rubber supplier. We conclude that the unsuspected presence of potential chemical depigmenting agents in solid bulk rubber, from which industrial rubber products are formulated, may contribute to the occurrence of occupational vitiligo, and that a simple review of ingredients in rubber formulations is inadequate to detect their presence.

Adult↗
PubMed1990

The Enzymatic Synthesis of Rubber Polymer in Parthenium argentatum Gray.

Washed rubber particles isolated from stem homogenates of Parthenium argentatum Gray by ultracentrifugation and gel filtration on columns of LKB Ultrogel AcA34 contain rubber transferase which catalyzes the polymerization of isopentenyl pyrophosphate into rubber polymer. The polymerization reaction requires Mg(2+) isopentenyl pyrophosphate, and an allylic pyrophosphate. The K(m) values for Mg(2+), isopentenyl pyrophosphate, and dimethylallyl pyrophosphate were 5.2 x 10(-4) molar, 8.3 x 10(-5) molar, and 9.6 x 10(-5) molar, respectively. The molecular characteristics of the rubber polymer synthesized from [(14)C]isopentenyl pyrophosphate were examined by gel permeation chromatography on three linear columns of 1 x 10(6) to 500 Angstroms Ultrastyragel in a Waters 150C Gel Permeation Chromatograph. The peak molecular weight of the radioactive polymer increased from 70,000 in 15 minutes to 750,000 in 3 hours. The weight average molecular weight of the polymer synthesized over a 3 hour period was 1.17 x 10(6) compared to 1.49 x 10(6) for the natural rubber polymer extracted from the rubber particles. Over 90% of the in vitro formation of the rubber polymer was de novo from dimethylallyl pyrophosphate and isopentenyl pyrophosphate. Treatment of the washed rubber particles with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate solubilized the rubber transferase. The solubilized enzyme(s) catalyzed the polymerization of isopentenyl pyrophosphate into rubber polymer with a peak molecular weight of 1 x 10(5) after 3 hours of incubation with Mg(2+) and dimethylallyl pyrophosphate. The data support the conclusion that the soluble preparation of rubber transferase is capable of catalyzing the formation of a high molecular weight rubber polymer from an allylic pyrophosphate initiator and isopentenyl pyrophosphate monomer.

Journal Article↗
PubMed2006

Magnesium ion regulation of in vitro rubber biosynthesis by Parthenium argentatum Gray.

Natural rubber is produced by a rubber transferase (a cis-prenyltransferase). Rubber transferase uses allylic pyrophosphate to initiate the rubber molecule and isopentenyl pyrophosphate (IPP) to form the polymer. Rubber biosynthesis also requires a divalent metal cation. Understanding how molecular weight is regulated is important because high molecular weight is required for high quality rubber. We characterized the in vitro effects of Mg(2+) on the biosynthetic rate of rubber produced by an alternative natural rubber crop, Parthenium argentatum (guayule). The affinity of the rubber transferase from P. argentatum for IPP.Mg was shown to depend on the Mg(2+) concentration in a similar fashion to the H. brasiliensis rubber transferase, although to a less extreme degree. Also, in vitro Mg(2+) concentration significantly affects rubber molecular weight of both species, but molecular weight is less sensitive to Mg(2+) concentration in P. argentatum than in H. brasiliensis.

Asteraceae↗
PubMed2000

Use of rubber dam and irrigant selection in UK general dental practice.

AIM: To evaluate factors which influence rubber dam use and irrigant selection in UK National Health Service (NHS) endodontics. METHODOLOGY: A postal survey was conducted amongst two age cohorts of dentists, representing all of the 1970-73 (older) and 1990-93 (younger) graduates of two northern English dental schools (n = 643). Key and supplementary questions were posed on levels of rubber dam use, irrigant selection, and factors influencing practice in NHS endodontics. After manual checking, validated (dual) entry of responses was made to a flat ASCII data file before analysis with SPSS software. The threshold for statistical significance was set at the 95% probability level. RESULTS: Eighty-five per cent of the valid sample responded to the questionnaire. Regardless of age and qualifying school, less than one-fifth of dentists always or frequently used rubber dam, whilst 60% never used it. Qualifying school had a significant influence on rubber dam use, whilst age had a variable influence. Major disincentives to the use of rubber dam included the perception that patients do not like it, that the NHS fee was inadequate to justify its use, that it took too long to apply, and that dentists had received inadequate training. Frequent users of rubber dam were significantly less likely to cite these disincentives than nonusers. Overall, local anaesthetic solution was the most common endodontic irrigant. Irrigant choice was strongly linked to rubber dam use, and to graduation cohort. Seventy-one per cent of rubber dam users irrigated with sodium hypochlorite, compared with only 38% of nonusers. This pattern was reversed for local anaesthetic irrigation. Younger graduates were significantly more likely to irrigate with local anaesthetic solution than their older counterparts, and the younger graduates of one school showed a highly significant increase in the use of chlorhexidine. CONCLUSIONS: 1) The majority of UK Health Service dentists never use rubber dam isolation in endodontic treatment. 2) Qualifying school has a significant impact on rubber dam use, and irrigant selection. 3) Use of rubber dam has a significant association with irrigant choice in endodontics.

Age Factors↗
PubMed1991

Frequency, spectrum and occupational relevance of type IV allergies to rubber chemicals.

3851 consecutive patients patch tested between January 1985 and March 1990 have been analysed for rubber allergies. The incidence of rubber allergy was 3.8% (n = 145). In 80/145 patients (55%), the source of rubber sensitization was occupational, 67 of whom (84%) had acquired allergy from wearing rubber gloves at work. Most of them (36%) were employed in the health services. The most commonly positive rubber-mix in this group was thiuram-mix (72%) followed by carba-mix (25%). 13/80 patients (16%) had occupational rubber allergy from industrial rubber products other than gloves. Patch tests revealed thiuram-mix (62%) as the most commonly positive rubber-mix but, in contrast to the group with glove-induced rubber allergy, black-rubber-mix came second (38%). In 47/145 patients (32%), the source of rubber sensitization was non-occupational; in 18/145 (13%) the origin remained unknown.

Adolescent↗
PubMed1996

Allergic patch test reactions caused by the rubber chemical cyclohexyl thiophthalimide.

Rubber chemicals are among the most common occupational contact sensitizers. The most common rubber sensitizers are thiurams, thiazoles, carbamates and paraphenylenediamine derivatives. Here we present data on a less-well-known rubber chemical, N-(cyclohexylthio)phthalimide (CTP; CAS 17796-82-6). This chemical is currently the most widely used vulcanization retarder, but data on allergic contact dermatitis caused by CTP are lacking. We conducted a survey of 310 patients who had been patch tested with 30 rubber chemicals including CTP. 11 (3.5%) showed an allergic patch test reaction provoked by CTP, and 9.0% by thiurams. 4 of the patients reacted only to CTP and not to other rubber chemicals, whereas the other 7 concomitantly reacted to other rubber chemicals. After analyzing the patch test data of these 11 patients, it was concluded that CTP probably did not cross-react with the other rubber chemicals. Therefore the patch test results may indicate independent sensitization to CTP and other rubber chemicals. Because very little data on the components of rubber chemicals in rubber products are available, the source of the putative sensitization to the rubber vulcanization retarder CTP is unknown.

Cross Reactions↗
PubMed1989

Allergic reactions to rubber condoms.

With the increased use of condoms, contact dermatitis to rubber is being seen more often. To develop a rubber condom suitable for use by rubber sensitive people, a "hypoallergenic" condom, which is washed in ammonia to reduce the residues of rubber accelerators, has been manufactured. Fifty patients allergic to various rubber accelerators were patch tested with an ordinary condom and the new washed condom. Fifty patients undergoing routine patch test investigation who were not allergic to rubber were also tested as controls. Twenty two of the rubber sensitive patients had a positive reaction to the new rubber condom compared with four of the control patients. Washing rubber condoms in ammonia does not appear to reduce the residues of rubber accelerators sufficiently for their use by rubber sensitive people. A non-allergenic condom is required.

Adolescent↗
PubMed1997

Natural rubber-containing medical devices; user labeling--FDA. Final rule.

The Food and Drug Administration (FDA) is issuing a final rule requiring labeling statements on medical devices, including device packaging containing natural rubber that contacts humans. The rule requires labeling of medical devices containing natural rubber latex that contacts humans to state: "Caution: This Product Contains Natural Rubber Latex Which May Cause Allergic Reactions."; labeling of medical devices containing dry natural rubber that contacts humans to state: "This Product Contains Dry Natural Rubber."; labeling of medical devices containing natural rubber latex in their packaging that contacts humans to state: "Caution: The Packaging of This Product Contains Natural Rubber Latex Which May Cause Allergic Reactions."; labeling of medical devices containing dry natural rubber in their packaging that contacts humans to state: "The Packaging of This Product Contains Dry Natural Rubber."; and that the claim of hypoallergenicity be removed from the labeling of medical devices that contain natural rubber. These requirements are being established in response to numerous reports of severe allergic reactions and deaths related to a wide range of medical devices containing natural rubber.

Dermatitis, Contact↗
PubMed2000

Alternative sources of natural rubber.

Rubber (cis-1,4-polyisoprene) is one of the most important polymers naturally produced by plants because it is a strategic raw material used in more than 40,000 products, including more than 400 medical devices. The sole commercial source, at present, is natural rubber harvested from the Brazilian rubber tree, Hevea brasiliensis. Primarily due to its molecular structure and high molecular weight (> 1 million daltons) this rubber has high performance properties that cannot easily be mimicked by artificially produced polymers, such as those derived from, e.g., bacterial poly-hydroxyalkanoates (PHAs). These high performance properties include resilience, elasticity, abrasion resistance, efficient heat dispersion (minimizing heat build-up under friction), and impact resistance. Medical rubber gloves need to fit well, be break-resistant, allow the wearer to retain fine tactile sensation, and provide an effective barrier against pathogens. The sum of all these characteristics cannot yet be achieved using synthetic gloves. The lack of biodiversity in natural rubber production renders continuity of supply insecure, because of the risk of crop failure, diminishing acreage, and other disadvantages outlined below. A search for alternative sources of natural rubber production has already resulted in a large number of interesting plants and prospects for immediate industrial exploitation of guayule (Parthenium argentatum) as a source of high quality latex. Metabolic engineering will permit the production of new crops designed to accumulate new types of valued isoprenoid metabolites, such as rubber and carotenoids, and new combinations extractable from the same crop. Currently, experiments are underway to genetically improve guayule rubber production strains in both quantitative and qualitative respects. Since the choice for gene activities to be introduced or changed is under debate, we have set up a complementary approach to guayule with yeast species, which may more quickly show the applicability and relevance of genes selected. Although economic considerations may prevent commercial exploitation of new rubber-producing microorganisms, transgenic yeasts and bacteria may yield intermediate or alternative (poly-)isoprenes suitable for specific applications.

Amino Acid Sequence↗
PubMed1994

Extractable latex allergens and proteins in disposable medical gloves and other rubber products.

BACKGROUND: IgE-mediated sensitization to rubber proteins is being reported with increasing frequency in health care workers. To explore the relative importance of various sources of allergen exposure, we measured the total rubber allergen and protein levels in extracts of disposable rubber gloves and compared the allergen levels with those in extracts of other medical and consumer rubber products. METHODS: Rubber allergens were measured by inhibition immunoassay with a rubber glove extract as the solid-phase allergen and pooled plasma from five rubber-sensitized health care workers as the IgE antibody source. Proteins were measured by Ninhydrin assay. RESULTS: Among 71 lots of gloves tested, the extractable allergen and protein levels were significantly correlated and were appreciably higher in powdered gloves than in powder-free gloves. Allergen levels varied 3000-fold among gloves from different manufacturers and were higher in examination gloves than in surgical or chemotherapy gloves. Measurable allergen was found in 11 of 24 lots of "hypoallergenic" gloves tested. Allergen levels in toy balloons were comparable to those in powdered gloves; much lower allergen levels were measured in condoms and anesthesia rebreathing bags. CONCLUSIONS: The allergen content of disposable rubber gloves varies widely and is higher in powdered gloves than in powder-free gloves and higher in examination gloves than in surgical gloves. Hypoallergenic gloves may contain substantial amounts of IgE-binding proteins. Gloves and toy balloons appear to be more important sources of rubber allergens than the other rubber products tested.

Allergens↗
PubMed1999

Isolation, characterization, and functional analysis of a novel cDNA clone encoding a small rubber particle protein from Hevea brasiliensis.

Biochemical evidence reported so far suggests that rubber synthesis takes place on the surface of rubber particles suspended in the latex of Hevea brasiliensis. We have isolated and characterized a cDNA clone that encodes a protein tightly bound on a small rubber particle. We named this protein small rubber particle protein (SRPP). Prior to this study, this protein was known as a latex allergen, and only its partial amino acid sequence was reported. Sequence analysis revealed that this protein is highly homologous to the rubber elongation factor and the Phaseolus vulgaris stress-related protein. Southern and Northern analyses indicate that the protein is encoded by a single gene and highly expressed in latex. An allergenicity test using the recombinant protein confirmed that the cloned cDNA encodes the known 24-kDa latex allergen. Neither ethylene stimulation nor wounding changed the transcript level of the SRPP gene in H. brasiliensis. An in vitro rubber assay showed that the protein plays a positive role in rubber biosynthesis. Therefore, it is likely that SRPP is a part of the rubber biosynthesis machinery, if not the rubber polymerase, along with the rubber elongation factor.

Allergens↗
PubMed1986

Dermatitis and urticaria from rubber and plastic gloves.

The number and nature of allergic occupational glove dermatoses were analysed. 542 cases of allergic contact dermatosis were diagnosed during 1974-1983. Amongst these, 68 (12.5%) were caused by rubber or plastic gloves. 2 patients had contact urticaria due to rubber gloves. Gloves were the main cause of occupational allergic rubber eczema, inducing 63 (58.3%) of 108 rubber eczema cases. 38 of them had positive reactions to rubber chemicals and glove material, 14 to glove material only, and 11 to rubber chemicals. 5 cases of allergic eczema from plastic gloves were diagnosed, all due to polyvinyl chloride (PVC). 2 cases of contact urticaria from natural rubber gloves were diagnosed by a provocation test. Epicutaneous testing with material of natural rubber gloves and rubber chemicals was negative. The present study shows that allergy to rubber gloves is usual, while allergy to plastic gloves is rare. Thus, plastic gloves should be used, when possible. Patch testing with protective gloves should always be used when patients develop prolonged hand dermatitis and where the possibility of glove eczema exists.

Clothing↗
PubMedDate not supplied

The effect of rubber dam placement on the arterial oxygen saturation in dental patients.

This study assessed the effect of rubber dam placement on arterial blood oxygen saturation in dental patients; it also determined whether the effects are technique sensitive. The study group consisted of 28 ASA Class I patients who were randomly allocated to one of two groups: Group A--rubber dam isolation of the maxilla (from tooth #14 to #6) and Group B-rubber dam isolation of the mandible (from tooth #19 to #27). A pulse oximeter was used to detect arterial blood oxygen saturation in both groups. Each patient's oxygen saturation (Sp02) was recorded every 30 seconds for two minutes to establish a baseline. Group A subjects received local infiltration in the vestibule above tooth #14, while Group B subjects received an inferior alveolar nerve block using 1.8 ml of 2% Lidocaine with 1:100,000 epiphrine, respectively. During the subsequent five minutes, the patient's Sp02 was recorded every 30 seconds. A rubber dam was then placed, which extended to the anterior septal angle (which completely covers the nose). This rubber dam remained in place for 20 minutes, with the patient's Sp02 being recorded every 30 seconds. The rubber dam was then altered (cut) to expose the nasal passages, creating what is known as proper rubber dam isolation, and the Sp02 was recorded every 30 seconds for 20 minutes. In both groups, there was no significant change in arterial oxygen saturation before or after rubber dam isolation was performed. Also, there was no significant difference in Sp02 when comparing the rubber dam isolation technique. Although rubber dam placement has no effect on blood oxygen levels in healthy patients, its effects on unhealthy patients are unknown.

Analysis of Variance↗
PubMed1998

In vitro and in vivo microbial adhesion and growth on argon plasma-treated silicone rubber voice prostheses.

Patients who undergo a total laryngectomy usually receive a silicone rubber voice prosthesis for voice rehabilitation. Unfortunately, biofilm formation on the esophageal side of voice prostheses limits their lifetime to 3-4 mon on average. The effects of repeated argon plasma treatment of medical grade, hydrophobic silicone rubber on in vitro adhesion and growth of bacteria and yeasts isolated from voice prostheses, as well as in vivo biofilm formation are presented here. In vitro experiments demonstrated that initial microbial adhesion over a 4 h time span to plasma-treated, hydrophilized, silicone rubber was generally less than on original, hydrophobic silicone rubber, both in the absence and presence of a salivary conditioning film on the biomaterial. Growth studies over a time period of 14 d at 37 degrees C in a modified Robbins device, showed that fewer Candida cells adhered on plasma-treated, hydrophilized silicone rubber as compared to on original, hydrophobic silicone rubber. For the in vivo evaluation of biofilm formation on plasma-treated silicone rubber voice prostheses, seven laryngectomized patients received a partly hydrophilized "Groningen Button" voice prosthesis for a planned evaluation period of 4 wk. After removal of the voice prostheses, the border between the hydrophilized and the original, hydrophobic side of the prostheses was clearly visible. However, biofilm formation was, unexpectedly, less on the original, hydrophobic sides, although the microbial compositions of the biofilms on both sides were not significantly different. Summarizing, this study demonstrates that in vitro microbial adhesion and growth on silicone rubber can be reduced by plasma treatment, but in vivo biofilm formation on silicone rubber voice prostheses is oppositely enhanced by hydrophilizing the silicone rubber surface. Nevertheless, from the results of this study the important conclusion can be drawn that in vivo biofilm formation on voice prostheses is controlled by the hydrophobicity of the biomaterials surface used.

Journal Article↗