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PubMed2004

Cinnamomin--a versatile type II ribosome-inactivating protein.

Ribosome-inactivating proteins (RIPs) are a group of toxic proteins that can specifically act on the universally conserved sarcin/ricin domain (S/R domain) of the largest RNA in ribosome and thus irreversibly inactivate ribosome for protein synthesis. Cinnamomin is a multifunctional type II RIP isolated in our laboratory from the mature seeds of the camphor tree. This protein has been extensively studied with regard to its purification, characteristics, structure and function, genetic expression, enzymatic mechanism, physiological role in seed cell and toxicity to cancer cells and insect larvae. The research results of cinnamomin obtained in our laboratory are summarized in this review. Understanding of cinnamomin and the relative new proteins will help expand our knowledge of RIPs and may accelerate theoretical study and the development of their potential applications.

Algal Proteins↗
PubMed2004

Cleavage of supercoiled circular double-stranded DNA induced by a eukaryotic cambialistic superoxide dismutase from Cinnamomum camphora.

A eukaryotic cambialistic superoxide dismutase (SOD) has been purified to homogeneity from mature seeds of the disease- and insect-resistant camphor tree (Cinnamomum camphora). Besides the known role of this SOD in protecting cells against oxidative stress, it can induce the cleavage of supercoiled double-stranded DNA into nicked and linear DNA. It can not cleave linear DNA or RNA, demonstrating there is no DNase or RNase in the purified cambialistic SOD. Furthermore, the SOD can linearize circular pGEM-4Z DNA that is relaxed by topoisomerase I. This result indicates that the DNA-cleaving activity requires substrates being topologically constrained. The supercoiled DNA-cleaving activity of the cambialistic SOD can be inhibited by either SOD inhibitor (azide) or catalase and hydroxyl radical scavengers (ethanol and mannitol). The chelator of iron, diethylenetriaminepentaacetic acid (DTPA), also inhibits the supercoiled DNA-cleaving activity. These results show that the dismutation activity is crucial for the supercoiled DNA cleavage. The modification of tryptophan residue of the cambialistic SOD with N-bromosuccinimide (NBS) shows that these two activities are structurally correlative. The reaction mechanism is proposed that the hydroxyl radical formed in a transition-metal-catalyzing Fenton-type reaction contributes to the DNA-cleaving activity. In addition, the cleavage sites in supercoiled pGEM-4Z DNA are random.

Catalase↗
PubMed1977

Clinical testing of olfaction reassessed.

Odours in current use for testing olfaction (e.g., peppermint, camphor) cause considerable trigeminal nerve stimulation; this would render them relatively inefficacious in the detection of lesions of the main olfactory pathway. Musks and floral odours are considered to be relatively 'pure' olfactory stimulants, acting virtually exclusively via the first cranial nerve. These odours have been compared with standard odours in a group of patients whose olfactory pathways had been damaged by frontal tumours, surgical operation, head injury, multiple sclerosis and miscellaneous causes. Hyposmia or anosmia was detected more frequently and more reliably by musks and floral odours in all groups of patients; a number of patients had gross deficits of odour description without hyposmia or anosmia. Two-thirds of an unselected group of patients with multiple sclerosis had olfactory abnormalities. A substantial improvement in the rate of detection of organic lesions affecting the olfactory pathway can be achieved by substituting odours such as musk ketone, exaltolide, linalyl acetate and coumarin for those in current use.

Humans↗
PubMed1995

Physiological and histological recovery of the olfactory mucosa of the frog after a dichlobenil injection.

Four days after a single systemic injection of 50 mg/kg of dichlobenil (2,6-dichlorobenzonitrile), the olfactory neuroepithelium of the frog is extensively damaged. At the same time, electrophysiological responses to odorant stimulations (2-heptanone, D-limonene, amyl acetate, camphor) are largely reduced. Pretreatment of the animals with metyrapone, an inhibitor of cytochrome P-450 enzymatic systems, inhibits the histological and physiological toxic effects of the dichlobenil injection. The olfactory tissue recovered 3 months after the dichlobenil injections and responses to odorant stimulations returned. The same dichlobenil injections did not induce lesions in the vomeronasal neuroepithelium.

Animals↗
PubMed1999

Olfactory discrimination ability of human subjects for ten pairs of enantiomers.

We tested the ability of human subjects to distinguish between enantiomers, i.e. odorants which are identical except for chirality. In a forced-choice triangular test procedure 20 subjects were repeatedly presented with 10 enantiomeric odor pairs and asked to identify the bottle containing the odd stimulus. We found (i) that as a group, the subjects were only able to significantly discriminate the optical isomers of alpha-pinene, carvone and limonene, whereas they failed to distinguish between the (+)- and (-)-forms of menthol, fenchone, rose oxide, camphor, alpha-terpineol, beta-citronellol and 2-butanol; (ii) marked individual differences in discrimination performance, ranging from subjects who were able to significantly discriminate between 6 of the 10 odor pairs to subjects who failed to do so with 9 of the 10 tasks; (iii) that with none of the 10 odor pairs were the antipodes reported to differ significantly in subjective intensity when presented at equal concentrations; and (iv) that error rates were quite stable and did not differ significantly between sessions, and thus, we observed a lack of learning or training effects. Additional tests of the degree of trigeminality and threshold measurements of the optical isomers of alpha-pinene, carvone and limonene suggest that the discriminability of these three enantiomeric odor pairs is indeed due to differences in odor quality. These findings support the assumption that enantioselective molecular odor receptors may only exist for some but not all volatile enantiomers and thus that chiral recognition of odorants may not be a general phenomenon but is restricted to some substances.

Adult↗
PubMed2006

Urinary excretion of morphine and codeine following the administration of single and multiple doses of opium preparations prescribed in Taiwan as "brown mixture".

Parallel to the "poppy-seed defense" strategy commonly reported in the United States, donors of urine samples tested positive for opiates in Taiwan often claimed the consumption of Brown Mixture (BM) as the source of the observed morphine and codeine. Because BM contains opium powder (10.0-10.5% morphine), opium tincture (0.9-1.1% morphine), or camphorated opium tincture (0.045-0.055% morphine) and is a popular remedy, and heroin use is considered a serious criminal act, the claim of BM use has to be adequately addressed. In this study, BM from seven different manufacturers (5 tablets and 2 solutions) and urine samples from alleged heroin users and volunteers with various ingestion patterns and were analyzed for their morphine and codeine contents. The analytical procedure included hydrolysis, trimethylsilylation, and gas chromatography-mass spectrometry analysis. The contents of morphine and codeine in the tablets were found to be very consistent, but with significant differences in the two BM solutions. Morphine concentrations found in urine specimens collected from volunteers ingesting BM tablets (or solutions) were always < 4000 ng/mL. The following morphine-to-codeine ([M]/[C]) ratios were observed for urine specimens with morphine concentration > or = 300 ng/mL: (A) < 3.0 for volunteers ingesting BM solution and (B) > 3.0 (mostly > 5.0) for volunteers ingesting BM tablets and alleged heroin users. It appeared that (A) BM ingestion (tablet or solution) was unlikely to result in a morphine concentration > 4000 ng/mL; and (B) [M]/[C] ratio might not be an effective parameter to differentiate heroin use from BM tablet ingestion.

Codeine↗
PubMed1983

Structural characteristics of cytochrome P-450. Possible location of the heme-binding cysteine in determined amino-acid sequences.

Computer-aided analyses were made of the complete amino-acid sequences of two P-450 species, the phenobarbital-inducible major P-450 of rat liver microsomes (P-450PB) and camphor-hydroxylating P-450 of Pseudomonas putida (P-450cam). Statistically significant homology was recognized between the two P-450 sequences, but these sequences were not related to those of other groups of hemoproteins, such as hemoglobins, peroxidases, and cytochrome c's and b's. Two highly homologous regions, HR1 and HR2, and two other weakly homologous regions were found on optimally matched alignment of the P-450 sequences. The secondary structures of the two P-450's predicted by current prediction methods bear strong resemblance at these homologous regions. Both HR1 and HR2 contain a cysteine residue near the center of the homologous regions, and they are the only regions that show significant homology among all 48 combinations of local sequences around the cysteine residues (six on P-450PB and eight on P-450cam). HR1 is located in the N-proximal half of the molecule, is rich in hydrophilic residues, and is predicted to be helical. On the other hand, HR2 is close to the C-terminus, has intermediate hydrophobicity, and may take a complex secondary structure of a turn-sheet-helix. The amino-acid sequences around the HR1 and HR2 regions are also well conserved in another P-450 species, rabbit P-450LM2.

Amino Acid Sequence↗
PubMed1992

Development of assays for the detection of photomutagenicity of chemicals during exposure to UV light. II. Results of testing three sunscreen ingredients.

Three sunscreen ingredients, derivatives of benzylidene camphor, were tested for photomutagenic potential. These were benzenesulfonic acid, 4-[(4,7,7,-trimethyl-3-oxo-bicyclo [2.2.1] hept-2-ylidene) methyl] (Mexoryl SL), 4-(2-oxo 3-bornylidenemethyl) phenyl trimethylammonium methyl sulphate (Mexoryl SO) and 3,3'-(1,4-phenylenedimethylidyne) bis [7,7-dimethyl-2-oxo-bicyclo [2.2.1] heptane-1-methanesulfonic acid] (Mexoryl SX). Two complementary assay systems were used, one involving the induction of reverse mutations in Escherichia coli strain WP2, the other measuring the induction of chromosome damage in Chinese hamster ovary (CHO) cells. Irradiation with UVA and/or UVB was provided by an Osram Ultra-Vitalux sunlamp. None of the three sunscreens, tested either to the limit of solubility or toxicity, gave any indication of photomutagenicity in either assay, under conditions in which the positive control compound, 8-methoxypsoralen, was extremely photomutagenic. It is concluded that Mexoryls SL, SO and SX can be exposed to UV light without producing photomutagenicity measurable using a bacterial reverse mutation or a mammalian chromosome aberration assay.

Animals↗
PubMed1995

Residues important for the function of a multihelical DNA binding domain in the new transcription factor family of Cam and Tet repressors.

We report that some prokaryotic repressors including CamR and TetR belong to the same family. CamR and TetR bind to DNA using a multihelical DNA binding domain (DBD) at the N-termini of the proteins, while the C-termini are important for regulating the DNA binding in a manner dependent on their co-factors (camphor for CamR, tetracycline for TetR). In all, 11 important amino acid positions have been identified in the CamR DBD by the systematic substitution of residues by Ala. Of the 11 positions, 10 are either buried in the core, and thus important for creating the hydrophobic environment, or exposed on the surface, and thus important for binding to DNA. The eleventh residue, Gly, seems to be important for a loop structure. The DNA binding mode of this type of DBD and a general mechanism of regulating their DNA binding are discussed in reference to the crystal structure of TetR [Hinrichs et al., (1994) Science, 264, 418-420].

Amino Acid Sequence↗
PubMed2006

Comparison of in vitro and in vivo estrogenic activity of UV filters in fish.

In this work, we evaluate whether in vitro systems are good predictors for in vivo estrogenic activity in fish. We focus on UV filters being used in sunscreens and in UV stabilization of materials. First, we determined the estrogenic activity of 23 UV filters and one UV filter metabolite employing a recombinant yeast carrying the estrogen receptor of rainbow trout (rtERalpha) and made comparisons with yeast carrying the human hERalpha for receptor specificity. Benzophenone-1 (BP1), benzophenone-2 (BP2), 4,4-dihydroxybenzophenone, 4-hydroxybenzophenone, 2,4,4-trihydroxy-benzophenone, and phenylsalicylate showed full dose-response curves with maximal responses of 81-115%, whereas 3-benzylidene camphor (3BC), octylsalicylate, benzylsalicylate, benzophenone-3, and benzophenone-4 displayed lower maximal responses of 15-74%. Whereas the activity of 17beta-estradiol was lower in the rtERalpha than the hERalpha assay, the activities of UV filters were similar or relatively higher in rtERalpha, indicating different relative binding activities of both ER. Subsequently, we analyzed whether the in vitro estrogenicity of eight UV filters is also displayed in vivo in fathead minnows by the induction potential of vitellogenin after 14 days of aqueous exposure. Of the three active compounds in vivo, 3BC induced vitellogenin at lower concentrations (435 microg/l) than BP1 (4919 microg/l) and BP2 (8783 microg/l). The study shows, for the first time, estrogenic activities of UV filters in fish both in vitro and in vivo. Thus we propose that receptor-based assays should be used for in vitro screening prior to in vivo testing, leading to environmental risk assessments based on combined, complementary, and appropriate species-related assays for hormonal activity.

Animals↗
PubMed1998

Review of the effectiveness of capsaicin for painful cutaneous disorders and neural dysfunction.

BACKGROUND: Topical capsaicin is known to be a safe and effective pain management adjunct for rheumatoid arthritis, osteoarthritis, neuralgias, and diabetic neuropathy. However, studies and case reports in the literature have indicated that other conditions may also benefit from capsaicin: painful or itching cutaneous disorders from operations, injuries, or tumors; neural dysfunction; or inflammation of the airways and urinary tract. METHODS: To determine the effectiveness of capsaicin for painful cutaneous disorders and neural dysfunction, the authors analyzed data from 33 reports (MEDLINE search of 1966-96) on the efficacy of capsaicin. Outcome measures consisted of the response rate and degree of pain relief. Results from placebo-controlled trials were pooled when possible; effect of treatment was estimated by the method of DerSimonian and Laird. RESULTS: Pain relief for postmastectomy syndrome and cluster headache was greater with capsaicin than with placebo; also, psoriasis and pruritus responded better to capsaicin. Uncontrolled studies and case reports have indicated that pain or dysfunction was less at the end of capsaicin therapy for neck pain, loin pain/hematuria syndrome, oral mucositis, rhinopathy, reflex sympathetic dystrophy syndrome, detrusor hyperreflexia, and cutaneous pain due to tumor of the skin. CONCLUSIONS: Capsaicin is effective for psoriasis, pruritus, and cluster headache; it is often helpful for the itching and pain of postmastectomy pain syndrome, oral mucositis, cutaneous allergy, loin pain/hematuria syndrome, neck pain, amputation stump pain, and skin tumor; and it may be beneficial for neural dysfunction (detrusor hyperreflexia, reflex sympathetic dystrophy, and rhinopathy). A universal problem for many of the studies analyzed was the absence of a "burning placebo" such as camphor.

Capsaicin↗
PubMed2001

Two methods to evaluate the antimicrobial action of calcium hydroxide paste.

The objective of this study was to analyze two methods for determining the antimicrobial effectiveness of (i) calcium hydroxide plus saline, (ii) calcium hydroxide plus polyethylene glycol, and (iii) calcium hydroxide plus camphorated paramonochlorophenol. Four microorganisms (Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 29212), Pseudomonas aeruginosa (ATCC 27853), and Bacillus subtilis (ATCC 6633)), one yeast (Candida albicans (ICB/USP-562)), and one mixture of these organisms were used. The strains were inoculated in Brain Heart Infusion (BHI) and incubated at 37 degrees C for 24 h. Two methods, the direct exposure test and the agar diffusion test were used to evaluate antimicrobial effects. For the direct exposure test (DET) 288 paper points were contaminated with the standard microbial suspensions and exposed to the intracanal dressings for 1, 24, 48, and 72 h. The points were immersed in Letheen Broth, followed by incubation at 37 degrees C for 48 h. An inoculum of 0.1 ml obtained from Letheen Broth was then transferred to 7 ml of BHI under identical incubation conditions, and microbial growth was evaluated. Pastes showed activity between 1 and 72 h, depending on the microorganism/mixture tested. For the agar diffusion test 36 Petri plates with 20 ml of BHI agar were inoculated with 0.1 ml of the same microbial suspension used for the DET, using sterile swabs that were spread on the medium. Five cavities were made in each of two agar plates (total = 10) and completely filled with one of the calcium hydroxide pastes. The plates were preincubated for 1 h at environmental temperature and then incubated at 37 degrees C for 24 to 48 h. The inhibition zone around each well was recorded in millimeters, and the results were submitted to ANOVA and Tukey test (alpha = 0.05). All intracanal dressings induced inhibition zones (range 5.0-10.0 mm). Data obtained showed that both the DET and agar diffusion test are useful in establishing the calcium hydroxide antimicrobial spectrum, thus improving infection control protocols. The direct exposure method is independent of other variables and is a practical laboratory procedure. A complete antimicrobial effect was observed after 48 h on indicator microorganisms, in both tests, irrespective of the calcium hydroxide paste vehicle.

Analysis of Variance↗
PubMed2002

Effectiveness of intracanal irrigants and medications against the yeast Candida albicans.

An in vitro study was conducted to determine the susceptibility of the yeast Candida albicans to various intracanal irrigants and medications. The minimum inhibitory concentration (MIC) of sodium hypochlorite, hydrogen peroxide, chlorhexidine digluconate, and aqueous calcium hydroxide that is required to kill a standardized inoculum of C. albicans was determined. Growth of the yeast was measured by optical density. Sodium hypochlorite, hydrogen peroxide, and chlorhexidine digluconate were effective anticandidal agents with MICs of <10 microgram/ml, 234 microgram/ml, and <0.63 microgram/ml, respectively. Aqueous calcium hydroxide had no activity. A standardized inoculum of C. albicans cells was also placed in direct contact with either calcium hydroxide paste or camphorated para-monochlorophenol (CPMC), and candidal growth was assessed by colony counts on Sabouraud's dextrose agar. Calcium hydroxide paste and CPMC, when maintained in direct contact with C. albicans, were effective antifungal agents.

Analysis of Variance↗
PubMed1990

Collodion as a safe, cost-effective dressing for central venous catheters.

Catheter-related sepsis continues to be a major problem with the use of central venous catheters. Controversy exists with respect to dressing material and frequency of dressing changes. Collodion is a solution of pyroxylin in a solution of 75% ether and 25% alcohol. Camphor and castor oil are added to create a flexible noncontracting dressing when applied to the skin. We retrospectively reviewed the charts of 34 patients requiring central venous catheters between 1986 and 1988. All catheters were placed via the subclavian approach. Collodion was used as a dressing on all patients. Dressings were not routinely changed. The catheters remained in position an average of 16.5 days. No insertion site became infected; one episode of catheter-related sepsis occurred, and two catheters were inadvertently dislodged. The overall incidence of catheter-related sepsis was 2.9%.

Bacterial Infections↗
PubMed1994

Transposon-mediated mobilization of chromosomally located catabolic operons of the CAM plasmid by TOL plasmid transposon Tn4652 and CAM plasmid transposon Tn3614.

The CAM (camphor degradation) plasmid is integrated into the chromosome of Pseudomonas putida PaW-line strains and is not self-transferable as a plasmid via conjugation. Our results show that the mobilization of chromosomally located CAM and the integration of cam-operons into the chromosome of the new Cam+ transconjugants is a recA-independent process mediated by transposons Tn4652 (17 kbp) and Tn3614 (7.2 kbp). Transposon Tn3614 is apparently identical to the left-hand and the right-hand sequences of the TOL plasmid pWW0 transposon Tn4654. The insertion of Tn401 inside the left-hand terminal IR of Tn4652 completely inhibited the mobilization of CAM. According to our data transposons Tn4652 and Tn3614 together with CAM plasmid catabolic operons are integrated into the chromosome. We propose that in pseudomonads the transposons Tn4652 and Tn3614 play a key role in the evolution and spread of new catabolic plasmids in nature.

Chromosomes, Bacterial↗
PubMed2002

Dynamics of low anisotropy morphologies in directional solidification.

We report experimental results on quasi-two-dimensional diffusion limited growth in directionally solidified succinonitrile with small amounts of poly(ethylene oxide), acetone, or camphor as a solute. Seaweed growth, or dense branching morphology, is selected by growing grains close to the [111] plane, where the in-plane surface tension is nearly isotropic. The observed growth morphologies are very sensitive to small anisotropies in surface tension caused by misorientations from the [111] plane. Different seaweed morphologies are found, including the degenerate, the stabilized, and the strongly tilted seaweeds. The degenerate seaweeds show a limited fractal scaling range and, with increased undercooling, suggests a transition from "fractal" to "compact" seaweed. Strongly tilted seaweeds demonstrate a significant twofold anisotropy. In addition, seaweed-dendrite transitions are observed in low anisotropy growth.

Journal Article↗
PubMed2005

Double dendrite growth in solidification.

We present experiments on the doublon growth morphology in directional solidification. Samples used are succinonitrile with small amounts of poly(ethylene oxide), acetone, or camphor as the solute. Doublons, or symmetry-broken dendrites, are generic diffusion-limited growth structures expected at large undercooling and low anisotropy. Low anisotropy growth is achieved by selecting a grain near the {111} plane leading to either seaweed (dense branching morphology) or doublon growth depending on experimental parameters. We find selection of doublons to be strongly dependent on solute concentration and sample orientation. Doublons are selected at low concentrations (low solutal undercooling) in contrast to the prediction of doublons at large thermal undercooling in pure materials. Doublons also exhibit preferred growth directions and changing the orientation of a specific doublonic grain changes the character and stability of the doublons. We observe transitions between seaweed and doublon growth with changes in concentration and sample orientation.

Journal Article↗
PubMed2003

Solid state solvation in amorphous organic thin films.

The photoluminescence (PL) of the red laser dye DCM2, doped into blended thin films of polystyrene (PS) and the polar small molecule camphoric anhydride (CA), redshifts as the CA concentration increases. The DCM2 PL peaks at 2.20 eV (lambda=563 nm) for pure PS films and shifts to 2.05 eV (lambda=605 nm) for films with 24.5% CA (by mass). The capacitively measured electronic permittivity also increases from epsilon=2.4 to epsilon=5.6 with CA concentration. These results are consistent with the theory of solvatochromism developed for organic molecules in liquid solvents. To our knowledge, this work is the first application of a quantitative theory of solvation to organic molecules in amorphous thin films with continuously controllable permittivity, and demonstrates that "solid state solvation" can be used to predictably tune exciton energies in organic thin film structures.

Journal Article↗