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Adverse effects of bone wax in surgery of the orbit.

The goal of this article is to establish the incidence and scope of adverse reactions to bone wax in a large orbital surgical series. We report two patients with bone wax granulomas of the orbit as a remote surgical complication. These are the first reported cases of adverse reactions to bone wax in the ophthalmic literature. A chart review was conducted on all patients from the University of British Columbia Orbit Clinic that had surgery with temporary or permanent removal of orbital bone. Two patients with bone wax granulomas were identified. In one case, intraoperative cultures grew Staphylococcus aureus, confirming that the wax may indeed act as a nidus for infection. No cases of pseudoarthrosis have occurred. This syndrome of chronic granulomatous giant cell foreign body inflammation has characteristic clinical, radiologic, and histopathologic features. The literature regarding adverse reactions to bone wax is reviewed, and specific implications for orbital surgery are discussed.

Child, Preschool↗

Polyester wax: a new embedding medium for the histopathologic study of human temporal bones.

BACKGROUND: Celloidin and paraffin are the two common embedding mediums used for histopathologic study of the human temporal bone by light microscopy. Although celloidin embedding permits excellent morphologic assessment, celloidin is difficult to remove, and there are significant restrictions on success with immunostaining. Embedding in paraffin allows immunostaining to be performed, but preservation of cellular detail within the membranous labyrinth is relatively poor. OBJECTIVES/HYPOTHESIS: Polyester wax is an embedding medium that has a low melting point (37 degrees C), is soluble in most organic solvents, is water tolerant, and sections easily. We hypothesized that embedding in polyester wax would permit good preservation of the morphology of the membranous labyrinth and, at the same time, allow the study of proteins by immunostaining. METHODS: Nine temporal bones from individuals aged 1 to 94 years removed 2 to 31 hours postmortem, from subjects who had no history of otologic disease, were used. The bones were fixed using 10% formalin, decalcified using EDTA, embedded in polyester wax, and serially sectioned at a thickness of 8 to 12 mum on a rotary microtome. The block and knife were cooled with frozen CO2 (dry ice) held in a funnel above the block. Sections were placed on glass slides coated with a solution of 1% fish gelatin and 1% bovine albumin, followed by staining of selected sections with hematoxylin and eosin (H&E). Immunostaining was also performed on selected sections using antibodies to 200 kD neurofilament and Na-K-ATPase. RESULTS: Polyester wax-embedded sections demonstrated good preservation of cellular detail of the organ of Corti and other structures of the membranous labyrinth, as well as the surrounding otic capsule. The protocol described in this paper was reliable and consistently yielded sections of good quality. Immunostaining was successful with both antibodies. CONCLUSION: The use of polyester wax as an embedding medium for human temporal bones offers the advantage of good preservation of morphology and ease of immunostaining. We anticipate that in the future, polyester wax embedding will also permit other molecular biologic assays on temporal bone sections such as the retrieval of nucleic acids and the study of proteins using mass spectrometry-based proteomic analysis.

Adolescent↗

Sandpipers (Scolopacidae) switch from monoester to diester preen waxes during courtship and incubation, but why?

Recently, a shift in preen wax composition, from lower molecular weight monoesters to higher molecular weight diesters, was described for individuals of a sandpiper species (red knot, Calidris canutus) that were about to leave for the tundra breeding grounds. The timing of the shift indicated that diester waxes served as a quality signal during mate choice. Here, this hypothesis is evaluated on the basis of a survey of preen wax composition in 19 sandpiper species. All of these species showed the same shift observed in the high-Arctic breeding red knots. As the shift also occurred in temperate breeding species, it is not specific to tundra-breeding sandpipers. Both sexes produced the diester waxes during the incubation period until hatching, in addition to the short period of courtship, indicating that diesters' functions extend beyond that of a sexually selected 'make-up'. The few non-incubating birds examined (males of curlew sandpipers (C. ferruginea) and ruffs (Philomachus pugnax)) had the lowest likelihood of secreting diesters, indicating a functional role for diester preen waxes during incubation. We propose that diester preen waxes enhance olfactory crypticism at the nest.

Animals↗

Mutation of the RESURRECTION1 locus of Arabidopsis reveals an association of cuticular wax with embryo development.

Insertional mutagenesis of Arabidopsis (Arabidopsis thaliana) was used to identify a novel recessive mutant, designated resurrection1 (rst1), which possesses a dramatic alteration in its cuticular waxes and produces shrunken nonviable seeds due to arrested embryo development. The RST1 gene sequence associated with these phenotypes was verified by three independent, allelic, insertion mutants, designated rst1-1, rst1-2, and rst1-3, with inserts in the first exon, 12th intron, and fourth exon, respectively. These three rst1 allelic mutants have nearly identical alterations in their wax profiles and embryo development. Compared to wild type, the wax on rst1 inflorescence stems is reduced nearly 60% in total amount, has a proportional reduction in aldehydes and aldehyde metabolites, and has a proportional increase in acids, primary alcohols, and esters. Compared to wild type, the C(29) alkanes on rst1 are nearly 6-fold lower, and the C(30) primary alcohols are 4-fold higher. These results indicate that rst1 causes shunting of most wax precursors away from alkane synthesis and into the primary-alcohol-producing branch of the pathway. In contrast to stems, the wax on rst1 mutant leaves increased roughly 43% in amount relative to the wild type, with the major increase occurring in the C(31) and C(33) alkanes. Unique among known wax mutants, approximately 70% of rst1 seeds are shrunken and nonviable, with these being randomly distributed within both inflorescence and silique. Viable seeds of rst1 are slightly larger than those of wild type, and although the viable rst1 seeds contain more total triacylglycerol-derived fatty acids, the proportions of these fatty acids are not significantly different from wild type. Shrunken seeds contain 34% of the fatty acids of wild-type seeds, with proportionally more palmitic, stearic, and oleic acids, and less of the longer and more desaturated homologs. Histological analysis of aborted rst1 seeds revealed that embryo development terminates at the approximate heart-shaped stage, whereas viable rst1 and wild-type embryos develop similarly. The RST1 gene encodes a predicted 1,841-amino acid novel protein with a molecular mass of 203.6 kD and a theoretical pI of 6.21. The RST1 transcript was found in all tissues examined including leaves, flowers, roots, stems, and siliques, but accumulation levels were not correlated with the degree to which different organs appeared affected by the mutation. The new RST1 gene reveals a novel genetic connection between lipid synthesis and embryo development; however, RST1's exact role is still quite unknown. The degree to which RST1 is associated with lipid signaling in development is an important focus of ongoing studies.

Arabidopsis↗

CER4 encodes an alcohol-forming fatty acyl-coenzyme A reductase involved in cuticular wax production in Arabidopsis.

A waxy cuticle that serves as a protective barrier against uncontrolled water loss and environmental damage coats the aerial surfaces of land plants. It is composed of a cutin polymer matrix and waxes. Cuticular waxes are complex mixtures of very-long-chain fatty acids and their derivatives. We report here the molecular cloning and characterization of CER4, a wax biosynthetic gene from Arabidopsis (Arabidopsis thaliana). Arabidopsis cer4 mutants exhibit major decreases in stem primary alcohols and wax esters, and slightly elevated levels of aldehydes, alkanes, secondary alcohols, and ketones. This phenotype suggested that CER4 encoded an alcohol-forming fatty acyl-coenzyme A reductase (FAR). We identified eight FAR-like genes in Arabidopsis that are highly related to an alcohol-forming FAR expressed in seeds of jojoba (Simmondsia chinensis). Molecular characterization of CER4 alleles and genomic complementation revealed that one of these eight genes, At4g33790, encoded the FAR required for cuticular wax production. Expression of CER4 cDNA in yeast (Saccharomyces cerevisiae) resulted in the accumulation of C24:0 and C26:0 primary alcohols. Fully functional green fluorescent protein-tagged CER4 protein was localized to the endoplasmic reticulum in yeast cells by confocal microscopy. Analysis of gene expression by reverse transcription-PCR indicated that CER4 was expressed in leaves, stems, flowers, siliques, and roots. Expression of a beta-glucuronidase reporter gene driven by the CER4 promoter in transgenic plants was detected in epidermal cells of leaves and stems, consistent with a dedicated role for CER4 in cuticular wax biosynthesis. CER4 was also expressed in all cell types in the elongation zone of young roots. These data indicate that CER4 is an alcohol-forming FAR that has specificity for very-long-chain fatty acids and is responsible for the synthesis of primary alcohols in the epidermal cells of aerial tissues and in roots.

Alcohols↗

The SHINE clade of AP2 domain transcription factors activates wax biosynthesis, alters cuticle properties, and confers drought tolerance when overexpressed in Arabidopsis.

The interface between plants and the environment plays a dual role as a protective barrier as well as a medium for the exchange of gases, water, and nutrients. The primary aerial plant surfaces are covered by a cuticle, acting as the essential permeability barrier toward the atmosphere. It is a heterogeneous layer composed mainly of lipids, namely cutin and intracuticular wax with epicuticular waxes deposited on the surface. We identified an Arabidopsis thaliana activation tag gain-of-function mutant shine (shn) that displayed a brilliant, shiny green leaf surface with increased cuticular wax compared with the leaves of wild-type plants. The gene responsible for the phenotype encodes one member of a clade of three proteins of undisclosed function, belonging to the plant-specific family of AP2/EREBP transcription factors. Overexpression of all three SHN clade genes conferred a phenotype similar to that of the original shn mutant. Biochemically, such plants were altered in wax composition (very long fatty acid derivatives). Total cuticular wax levels were increased sixfold in shn compared with the wild type, mainly because of a ninefold increase in alkanes that comprised approximately half of the total waxes in the mutant. Chlorophyll leaching assays and fresh weight loss experiments indicated that overexpression of the SHN genes increased cuticle permeability, probably because of changes in its ultrastructure. Likewise, SHN gene overexpression altered leaf and petal epidermal cell structure, trichome number, and branching as well as the stomatal index. Interestingly, SHN overexpressors displayed significant drought tolerance and recovery, probably related to the reduced stomatal density. Expression analysis using promoter-beta-glucuronidase fusions of the SHN genes provides evidence for the role of the SHN clade in plant protective layers, such as those formed during abscission, dehiscence, wounding, tissue strengthening, and the cuticle. We propose that these diverse functions are mediated by regulating metabolism of lipid and/or cell wall components.

Acclimatization↗

Age-related changes in sebaceous wax ester secretion rates in men and women.

Sustainable rates of sebaceous wax ester secretion were measured on the foreheads of 109 men and 167 women, aged 15-97. Each measurement was made after first depleting the cutaneous sebum reservoir by overnight absorption of lipid into a layer of bentonite clay. Lipid was then absorbed for 3 h into fresh clay in which two 2-cm cloth disks were embedded. The absorbed lipid was extracted from the disks with ether and analyzed for wax esters by thin-layer chromatography. For both men and women there was a wide range of wax ester secretion rates at all ages. Rates were highest in the 15- to 35-year-olds and appeared to decline continuously throughout the adult age range. Values of log(wax esters) were better correlated with age than the untransformed values of wax ester secretion. The equations of best fit of log(wax esters) vs age suggested that sebum secretion declines about 23% per decade in men and 32% per decade in women.

Adolescent↗

Periodic evaluation of the occlusal vertical dimension of maxillary dentures from the wax trial denture through 48 hours after polymerization.

PURPOSE: This study measured the effects of commercial resin type on maxillary complete dentures with monoplane teeth by periodically comparing the occlusal vertical dimension of the polymerized dentures with the baseline, wax trial denture fiducial measurements. MATERIALS AND METHODS: Commercially available compression-molded, injection-molded, and fluid poly(methyl methacrylate) resins, as well as one compression-molded methyl acrylate ester copolymer, were evaluated. Ten dentures were fabricated from each resin using monoplane teeth. The occlusal vertical dimension at the articulator pin was measured at the wax denture stage for each specimen, and changes in occlusal vertical dimension for each denture were evaluated at a simulated laboratory remount, and at 0, 3, 6, 24, and 48 hours after a simulated clinical remount. Repeated measures analysis of variance (alpha = 0.05) and post hoc one-way factorial analysis of variance and Scheffe's F-Tests for each resin group were performed using ranks of raw data. RESULTS: Changes in the maxillary denture mean occlusal vertical dimensions were recognized throughout the evaluation periods compared with the wax-denture baseline, and time was a significant influence on displacement (p = .0001). Only the compression-molded poly(methyl methacrylate) dentures exhibited a mean laboratory remount occlusal vertical dimension that was significantly greater than the mean wax denture measurement, and all resin systems exhibited occlusal error that was significantly less than the laboratory remount measurements at 48 hours. At 48 hours, all resin groups exhibited mean occlusal vertical dimension changes that were less than 1 mm compared with the wax denture. Only compression-molded poly(methyl methacrylate) dentures exhibited a mean 48-hour clinical remount measurement that was statistically similar to the mean wax denture occlusal vertical dimension. CONCLUSIONS: Individual maxillary dentures from all resin types and at all intervals exhibited dimensional change. At the last evaluation period, the compression-molded poly(methyl methacrylate) showed no change in mean occlusal vertical dimension from baseline, whereas remaining groups exhibited occlusal vertical dimensions significantly less than baseline.

Acrylates↗

Neutral lipid biosynthesis in engineered Escherichia coli: jojoba oil-like wax esters and fatty acid butyl esters.

Wax esters are esters of long-chain fatty acids and long-chain fatty alcohols which are of considerable commercial importance and are produced on a scale of 3 million tons per year. The oil from the jojoba plant (Simmondsia chinensis) is the main biological source of wax esters. Although it has a multitude of potential applications, the use of jojoba oil is restricted, due to its high price. In this study, we describe the establishment of heterologous wax ester biosynthesis in a recombinant Escherichia coli strain by coexpression of a fatty alcohol-producing bifunctional acyl-coenzyme A reductase from the jojoba plant and a bacterial wax ester synthase from Acinetobacter baylyi strain ADP1, catalyzing the esterification of fatty alcohols and coenzyme A thioesters of fatty acids. In the presence of oleate, jojoba oil-like wax esters such as palmityl oleate, palmityl palmitoleate, and oleyl oleate were produced, amounting to up to ca. 1% of the cellular dry weight. In addition to wax esters, fatty acid butyl esters were unexpectedly observed in the presence of oleate. The latter could be attributed to solvent residues of 1-butanol present in the medium component, Bacto tryptone. Neutral lipids produced in recombinant E. coli were accumulated as intracytoplasmic inclusions, demonstrating that the formation and structural integrity of bacterial lipid bodies do not require specific structural proteins. This is the first report on substantial biosynthesis and accumulation of neutral lipids in E. coli, which might open new perspectives for the biotechnological production of cheap jojoba oil equivalents from inexpensive resources employing recombinant microorganisms.

Acinetobacter↗

Mastoid canal and migrated bone wax in the sigmoid sinus: technical report.

A study of the migration of bone wax into the sigmoid sinus through the mastoid canal is reported here. In 7 of 161 patients who underwent retromastoid craniectomy, the postoperative soft tissue window image computed tomographic scans demonstrated a hypodense mass in the ipsilateral sigmoid sinus. The density value of the hypodense mass ranged from -34 to -79 Hounsfield units, which was neither as low as that of air nor as high as that of cerebrospinal fluid, but was comparable to that of fat tissue or bone wax. The continued presence of all of these masses in the sigmoid sinus was confirmed 1 month to 2 years after surgery. These computed tomographic findings suggested that this abnormal hypodense mass might be a migrated fragment of the bone wax that had been used for the control of venous bleeding from the mastoid emissary vein, because each of the seven affected patients had a large mastoid foramen and a large quantity of bone wax had been needed to control the bleeding during retromastoid craniectomy. No other material with the potential to migrate into the sigmoid sinus had been applied as a packing material. In two of the seven patients, venous magnetic resonance angiography after surgery demonstrated that the ipsilateral sigmoid sinus was not patent and the computed tomographic scans also revealed that the hypodense masses occupied the sigmoid sinus. It is concluded that the intrasurgical application of a large quantity of bone wax to control the bleeding from the large emissary veins carries a risk of the migration of bone wax into the sigmoid sinus.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Attachment forces of ants measured with a centrifuge: better 'wax-runners' have a poorer attachment to a smooth surface.

The symbiotic ant partners of glaucous Macaranga ant-plants show an exceptional capacity to run on the slippery epicuticular wax crystals covering the plant stem without any difficulty. We test the hypothesis that these specialised 'wax-runners' have a general, superior attachment capacity. We compared attachment on a smooth surface for 11 ant species with different wax-running capacities. The maximum force that could be withstood before an ant became detached was quantified using a centrifuge recorded by a high-speed video camera. This technique has the advantage of causing minimum disruption and allows measurements in very small animals. When strong centrifugal forces were applied, the ants showed a conspicuous 'freezing reflex' advantageous to attachment. Attachment forces differed strongly among the ant species investigated. This variation could not be explained by different surface area/weight ratios of smaller and larger ants. Within species, however, detachment force per body weight (F/W) scaled with the predicted value of W(-)(0.33), where W is body weight in newtons. Surprisingly, our results not only disprove the hypothesis that 'wax-runners' generally attach better but also provide evidence for the reverse effect. Superior 'wax-runners' (genera Technomyrmex and Crematogaster) did not cling better to smooth Perspex, but performed significantly worse than closely related congeners that are unable to climb up waxy stems. This suggests an inverse relationship between adaptations to run on wax and to attach to a smooth surface.

Animals↗

Spectroscopic assessment of Australian cotton waxes.

An investigation into the spectroscopic analysis of cotton waxes on Australian cottons was undertaken. The chemical composition of cotton wax is complex and contains a number of lipid classes. Infrared transmission spectroscopy coupled with principal component analysis was found to be capable of discriminating between solvent-extracted cotton waxes with differences in their alkyl functionality. Based on high-performance thin-layer chromatography (HPTLC) results, these differences were associated with an increase in levels of the alkane wax component. On the basis of these results, a photo-acoustic spectroscopic method was developed that could be used to distinguish raw cottons on the basis of these differences. This method was utilized to screen cottons from the Cotton Seed Distributors 2001 seed trial. A preliminary assessment of the scouring and dyeing properties of the various cottons, identified using the photo-acoustic method, was carried out. The results tended to confirm that cottons with increased alkyl functionality, most likely associated with alkane wax, were more difficult to remove and residual wax on the fiber acted as a barrier to dyestuff penetration, thus lowering color yield.

Australia↗

Epicuticular waxes from caatinga and cerrado species and their efficiency against water loss.

The effects of the contents and chemical composition of the foliar epicuticular waxes of species from the caatinga (Aspidosperma pyrifolium, Capparis yco, Maytenus rigida and Ziziphus joazeiro) and cerrado (Aristolochia esperanzae, Didymopanax vinosum, Strychnos pseudoquina and Tocoyena formosa) were evaluated as to the resistance to water loss by means of an experimental device constructed for this purpose. In general, the waxes of the caatinga species investigated were more efficient against water loss than cerrado species. Increase of the thickness of the waxy deposits from 40 to 90 microg.cm-2 had no significant effect on the resistance to water loss. The chemistry of the wax constituents was shown to be an important factor to determine the degree of resistance to evaporation. n-Alkanes and alcoholic triterpenes were the most efficient barriers, while hentriacontan-16-one (a ketone) and ursolic acid (an acid triterpene) revealed low efficiency. The higher efficiency of the waxes of the leaves from caatinga species (mainly those of C. yco and Z. joazeiro) is probably accounted for the predominance of n-alkanes in their composition. The lower efficiency of the waxes of A. pyrifolium (caatinga), T. formosa and A. esperanzae (both species from the cerrado) is probably a consequence of the predominance of triterpenoids in the waxes of the two former species and hentriacontan-16-one in the latter.

Brazil↗

Fabrication of a wax ear by rapid-process modeling using stereolithography.

PURPOSE: The purpose of this article is to describe an automated technique that creates a mirror-image wax model of the contralateral ear using a rapid prototyping technique, which allows precise positioning and adaptation to the deficient side of the face. MATERIALS AND METHODS: Magnetic resonance images (MRI) are taken of the face and then digitized and reformatted into a 3-dimensional image. The image of the normal ear is extracted, mirrored, and positioned onto the image of the deformed side of the face. The digitized data, when used in conjunction with stereolithography, produces a photopolymerized resin model ear. A model wax ear is created by pouring molten wax into a silicone mold of the resin model ear. RESULTS: A model wax ear of identical dimensions, shape, and internal contouring as the contralateral normal ear is created. The technique allows the undersurface of the model wax ear to be precisely positioned against the surface contours of the deformed side of the face in 1 patient. CONCLUSION: The use of stereolithography from MRI scan data is a suitable technique for producing a model wax ear that fits the deformed side of the face. The 3-dimensional computer image of the face may also be used to plan the position (level and protrusion) of the ear.

Computer-Aided Design↗

Waxed windshields are hazardous in the rain.

Seven automobiles were washed and waxed at four car washes. Photographic determinations were made of the glare produced by the wet or dry waxed windshield in a headlight beam. When wet, the waxed windshields scattered three times more light than in the normal human eye. The wet wax scattering was 24.8 times higher than when dry. No wax residues should be permitted on windshields and the National Highway Traffic Safety Administration should issue a mandatory windshield cleaning requirement after waxing.

Automobile Driving↗

Studies on the compressibility of wax matrix granules of acetaminophen and their admixtures with various tableting bases.

Matrix granules of acetaminophen have been formed by a melt granulation process whereby the acetaminophen powder was triturated with the melted wax--goat wax, glyceryl monostearate or carnuba wax. The compressibility of the matrix granules and their admixture, with diluent granules (lactose, alpha-cellulose or microcrystalline cellulose) was investigated. The granules were compressed to tablets at a constant load (30 arbitrary units on the load scale) of a manesty single punch machine. Resulting tablets were evaluated for tensile strength (T) and disintegration times (DT). Granule flow was determined by measuring their angle of repose when allowed to fall freely on a level surface. Matrix granules prepared by melt granulation with goat wax or glyceryl monostearate were too sticky and therefore did not flow at all. They were also poorly compressible (T values = 0.20MN/m2). Inclusion of the diluent remarkably improved granule flow property and compressibility. The T values of the tablets (measure of compressibility) increased from about 0.24 to 0.65 MN/m2 during increase in diluent (lactose) content from 20 to 80 %w/w. Microcrystalline cellulose and alpha-cellulose were more effective than lactose in promoting compressibility of the granules. By contrast the matrix granules formed with carnuba wax were free flowing (angle of repose, 18.60). Addition of the diluent further improved flowability slightly. The matrix granules (without a diluent) were readily compressible (T value, 1.79MN/m2). Addition of the diluent (80%w/w) reduced T values (MN/m2) slightly to 1.32 (lactose), 1.48 (alpha-cellulose) and 1.74 (microcrystalline cellulose). Tablets of the matrix granules only, disintegrated rapidly within 3 minutes. DT was further reduced to <30 s by addition of any of the diluents. The indication is that the inclusion of the diluents studied can be used to improve the compressibility of the otherwise poorly compressible matrix granules. Based on the flowability, compressibility, and disintegration data, carnuba wax proved most promising in the melt granulation of the test drug for sustained release applications.

Acetaminophen↗

[Plant epicuticular wax and drought resistance as well as its molecular biology].

Plant epicuticular wax is a collective term used to describe the organic components of the cuticle which covers the outer surface of aerial plant tissues. It is also the last defensive barrier of plant, playing important roles in plant growth and development. The components, transport, crystal formation and morpho-ultrastructure of plant epicuticular wax as well as the investigations concerning the relationship between epicuticular wax and adaptations to environment are reviewed. Especially, we systematically summarized the relationships among wax, cutin transpiration, leaf WUE and yield and the progresses in the physiological and molecular biology research regarding wax; we also discussed the problems existing in wax research.

Adaptation, Physiological↗

The promotional effect of bone wax on experimental Staphylococcus aureus osteomyelitis.

The consequences of using surgical bone wax are not well studied. We evaluated the infection-promoting potential of sterile bone wax in a rat model of chronic Staphylococcus aureus osteomyelitis. The addition of bone wax greatly reduced the quantitative bacterial inoculum (log colony-forming units) required to establish chronic osteomyelitis in 50% and 100% of challenged animals. The 50% infection rate was reduced from log 6.9 to 2.6 and the 100% infection rate from 8.2 to 4.4, respectively (p less than 0.015, t test for parallelism). Separate experiments were done 10 to 30 minutes after inoculation with only log 6.4 staphylococci. Tibiae of animals that received bone wax yielded more organisms than those that did not (log 2.76 +/- 0.68 versus 1.72 +/- 0.94, p less than 0.01). At 24 hours quantitative colony counts were not significantly different whether animals received wax or not (log 5.02 +/- 0.42 versus 4.43 +/- 0.65, p greater than 0.09). These studies suggest that the routine surgical use of bone wax should be reassessed.

Animals↗