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PubMed1991

Primary and secondary structural patterns in eukaryotic cytochrome P-450 families correspond to structures of the helix-rich domain of Pseudomonas putida cytochrome P-450cam. Indications for a similar overall topology.

An extensive sequence analysis of the eukaryotic cytochrome P-450 (P-450) protein families was conducted with a view to identifying conserved regions that might be related to secondary structural features in the Pseudomonas putida camphor hydroxylase (P-450cam). All sequences available on-line were collected, classified and aligned within families. Distinctively different sequences were chosen from each of seven eukaryotic families, and an unbiased multi-alignment was constructed. Profile patterns of the most conserved regions were generated and screened against the sequence of P-450cam, the structure of which has been elucidated by X-ray crystallography. While some of these profiles did not map on the P-450cam sequence, the structurally most important helices were clearly identified and the correlations were found to be statistically significant. Our analysis suggests that the helix-rich domain with the cysteine pocket and the oxygen-binding site is conserved in all P-450 forms. Helices I and L from P-450cam can be easily identified in all eukaryotic P-450 forms. Other helices which seem to exist in all P-450 forms include helices C, D, G and J. K. In the helix-poor domain of P-450cam, only structures b3/b4 seem to have been conserved. The obvious sequence conservation throughout the helix-rich domain of the P-450cam protein might be expected for a molecular class whose overall topology is preserved. Additional support for the conservation of structure between eukaryotic cytochromes P-450 and P-450cam comes from secondary structure prediction of the eukaryotic sequences.

Amino Acid Sequence↗
PubMed1994

Reaction of inflamed periapical tissue to intracanal medicaments and root canal sealers.

The effects of different intracanal medicaments and zinc oxide-eugenol based root canal sealers on an experimentally induced apical periodontitis were studied histologically in mesial roots of lower molars of Wistar rats. After root canal instrumentation each canal was filled either with an intracanal medicament twice for 3 days each time or with a sealer for 21 days. The intracanal medicament chlorophenol caused periapical tissue damage. P-chloroxylenol-camphor lead to periapical improvement as did 5% sodium-hypochlorite. 12% sodium-hypochlorite gave tissue damage. After intracanal medication with an aqueous suspension of calcium hydroxide periapical repair was clearly visible. A combination of a corticoid and an antibiotic appeared to induce damage of the periapical tissues. The root canal sealers Endomethasone and N2 essentially impaired periapical repair. The results were acceptable for Aptal-Zink-Harz root canal fillings after 21 days as well as after 56 days.

Administration, Topical↗
PubMed1980

The risk of jaundice in glucose-6-phosphate dehydrogenase deficient babies exposed to menthol.

A major cause of neonatal morbidity and mortality in Lagos, Nigeria, is severe neonatal jaundice seen in G-6-PD deficient babies. The observation that the jaundice is more severe in outpatient than in inpatient babies suggests that its cause is exogenous. "Mentholated" powder which is commonly used in many clinics and at home to dress umbilical cords was suspected to be the offending agent. A controlled study of the effects of one of these powders was carried out on 60 consecutive G-6-PD deficient babies. In 30 of them the umbilical cords were dressed daily with the powder while the remaining half who were untreated served as controls. The treated babies developed statistically more significant jaundice than the controls. Inability of neonates to conjugate menthol in this power is probably responsible for the jaundice developed by these G-6-PD deficient babies. It is concluded that the use of menthol and/or camphor-containing commerical products on neonates be discontinued, especially in communities where the incidence of G-6-PD deficiency is high as the use of such products may be contributiing to the severity of neonatal jaundice.

Bilirubin↗
PubMed1970

Cytochrome content of two pseudomonads containing mixed-function oxidase systems.

The cytochrome and nonheme iron protein content of two pseudomonads, Pseudomonas oleovorans and P. putida, containing mixed function oxidase systems was examined. The mixed function oxidase system of P. oleovorans and P. putida had previously been shown to be present in cells which had been grown on hexane and camphor, respectively, as energy source. The content of protoheme was found to increase significantly when the organisms were grown on the substrates for mixed function oxidation. The nonheme iron protein content increased significantly in the case of P. putida and was constant in P. oleovorans. The cytochrome c content was essentially constant in both pseudomonads. The content of cytochrome P-450 in P. putida increased from an immeasurably low amount to 0.15 nmoles per mg (dry weight). The content of cytochrome o in P. oleovorans increased by a factor of 4.5. P. oleovorans was not found to contain detectable quantities of cytochrome P-450 either in the presence or absence of the mixed function oxidase system.

Alkanes↗
PubMed1973

Common enzymes of branched-chain amino acid catabolism in Pseudomonas putida.

Two types of Pseudomonas putida PpG2 mutants which were unable to degrade branched-chain amino acids were isolated after mutagenesis and selection for ability to grow on succinate, but not valine, as a sole source of carbon. These isolates were characterized by growth on the three branched-chain amino acids (valine, isoleucine, and leucine), on the corresponding branched-chain keto acids (2-ketoisovalerate, 2-keto-3-methylvalerate, and 2-ketoisocaproate), and on other selected intermediates as carbon sources, and by their enzymatic composition. One group of mutants lost 2-ketoisovalerate-inducible branched-chain keto acid dehydrogenase that was active on all three keto acids. There was also a concomitant loss of ability to grow on all three branched-chain amino acids as well as on all three corresponding keto acids, but there was retention of ability to use subsequent intermediates in the catabolism of branched-chain amino acids. Another type of mutant showed a marked reduction in branched-chain amino acid transaminase activity and grew poorly at the expense of all three amino acids, but it utilized subsequent intermediates as carbon sources. Both the transaminase and branched-chain keto acid dehydrogenase mutants retained the ability to degrade camphor. These findings are consistent with the view that branched-chain amino acid transaminase and branched-chain keto acid dehydrogenase are common enzymes in the catabolism of valine, isoleucine, and leucine.

Alcohol Oxidoreductases↗
PubMed1977

Transposition of a beta-lactamase locus from RP1 into Pseudomonas putida degradative plasmids.

The beta-lactamase gene from the RP1 plasmid transposes into at least two Pseudomonas putida degradative plasmids. Donor strains that carry RP1 (bla+ tet+ aphA+) and a degradative plasmid yield transconjugants that have only the bla+ marker of RP1. This occurs in up to 80% of all bla+ transconjugants. Segregation of the bla+ marker requires the presence of a degradative plasmid in the donor and is only observed in transconjugants that have received degradative markers. The bla+ tet aphA transconjugants show 100% linkage of bla+ to degradative markers in conjugation,transduction, and transformation crosses. A transduction cross of an (RP1), (SAL) donor shows that 8% of all SAL plasmids also carry the transposed bla+ marker. Tn401 is the name we assign to the bla+ transposon from RP1 observed in Pseudomonas. Its identity with the RP1 bla+ transposon observed in Escherichia coli is not known. In four cases, Tn401 has inserted into the camphor genes of the CAM-OCT plasmid.

Alkanes↗
PubMed1993

Evidence for autoregulation of camR, which encodes a repressor for the cytochrome P-450cam hydroxylase operon on the Pseudomonas putida CAM plasmid.

The regulatory gene camR on the CAM plasmid of Pseudomonas putida (ATCC 17453) negatively controls expression of the cytochrome P-450cam hydroxylase operon (camDCAB) for the camphor degradation pathway and is oriented in a direction opposite to that of the camDCAB operon. In this study, we examined expression of the camR gene by monitoring the beta-galactosidase activity of camR-lacZ translational fusions in P. putida camR and camR+ strains. We found that the camR gene was autogenously regulated by its own product, CamR. To search for an operator site of the camR gene, a cam repressor (CamR)-overproducing plasmid, pHAOV1, was constructed by placing the camR gene under the control of a pL promoter. The translational initiation codon of CamR was changed by site-directed mutagenesis from GTG to ATG to improve translation efficiency. Judging from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the CamR protein was expressed up to about 10% of the soluble protein of CamR-overproducing Escherichia coli JM83/pHAOV1 cells. Results of DNase I footprinting assays using the cell lysate indicated that the CamR repressor covered a single region between the camR gene and the camDCAB operon. Our findings also suggest that the camR gene autogenously regulates its own expression by binding of the gene product, CamR, to the operator, which also serves as an operator of the camDCAB operon.

Base Sequence↗
PubMed2002

Comparative investigation of the antimicrobial activity of PADMA 28 and selected European herbal drugs.

OBJECTIVE: PADMA 28 is a multicompound preparation of 20 herbs, calcium sulphate, and camphor, derived from Tibetan medicine. It is usually used in the treatment of peripheral circulatory disorders, accompanied by the symptoms tingling, formication, heaviness and tenseness in arms and legs, numbness in hands and feet, and cramps in the calf. Recently, the question of whether appropriate preparations of PADMA 28 also exhibit antibacterial and antimycotic activity has often been raised. As there are as yet no experimental findings that answer this question, an in vitro study was carried out. In a parallel survey we investigated the antimicrobial properties of 5 herbal drugs which are commonly used in the traditional European folk medicine for the topical treatment of mild skin infections, wounds and eczematous skin lesions. METHODS: The minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) of alcohol-based (tinctures) and aqueous (teas) herbal drug preparations were determined in vitro by a broth microdilution method for 5 Gram-positive and 5 Gram-negative bacteria, as well as the yeast Candida albicans. RESULTS: The aqueous and alcohol-based PADMA 28 preparations as well as the corresponding preparations of the European herbal drugs showed an antibacterial effect against Gram-positive bacteria in vitro. These bacteria revealed a somewhat higher sensitivity to the teas prepared from the European herbal drugs (MIC: 1.3-20.0 mg/ml) than to the aqueous preparations of PADMA 28 (MIC: 5.0-40.0 mg/ml). The better antibacterial activity of the European herbal drugs is probably based on their relatively high amount of tanning agents. On the other hand, all tested plant preparations inhibited not at all or only insufficiently the growth of the Gram-negative bacteria tested and that of Candida albicans. The ethanolic PADMA 28 tinctures showed an improved inhibitory effect on the Gram-positive bacteria (MIC: 0.38-1.51% tincture or 0.38-1.51 mg PADMA 28/ml) compared with the aqueous preparations; this effect is comparable to the ethanolic tinctures of the tested European herbal drugs (MIC: 0.4-1.6/3.2% tincture or 0.4-1.6/3.2 mg herbal drug/ml). CONCLUSION: All tested tea preparations and alcoholic tinctures of PADMA 28 as well as those of the selected European herbal drugs exhibited evident antibacterial effects against Gram-positive bacteria in vitro. On the other hand, except for Klebsiella pneumoniae, all Gram-negative bacteria tested and the yeast Candida albicans were insensitive against the different aqueous and alcohol-based plant extracts.

Anti-Bacterial Agents↗
PubMedDate not supplied

Afferent signals to the CNS appear not to condition the modulation of interleukin-1 receptors in the hippocampus.

Conditioned alteration of natural killer (NK) cell activity was used as an indicator of the functional bidirectional communication between the immune and central nervous systems. Poly I:C and lipopolysaccharide (LPS) from Escherichia coli and Salmonella typhimurium were used as unconditioned stimuli and odor of camphor as the conditioned stimulus. An attempt was made to demonstrate the role of central interleukin (IL-1) receptors in this communication process. Brain IL-1 receptors were down-regulated by treatment with 50 microg/mouse of LPS from S. typhimurium, but not with the same dose of LPS from E. coli or poly I:C. A significant level of conditioned augmentation of NK cell activity was observed with poly I:C. Conditioned alteration in NK cell activity was also observed with LPS from E. coli, but at much lower level than poly I:C. NK cell activity was not conditioned with LPS from S. typhimurium at the same dose as E. coli LPS, but conditioned enhancement of NK cell activity was observed with a higher dose (100 microg) of S. typhimurium LPS. These results suggest that modulation of central IL-1 receptors do not seem to play a role in the conditioned augmentation of NK cell activity.

Afferent Pathways↗
PubMed1980

Evaluation of surfactant-like effects of commonly used remedies for colds.

Volatile aromatics, eucalyptol, eucalyptus oil, camphor and menthol were spread on synthetic DPL films and pulmonary surfactants; the initial surface pressure of the surfactants was measured and the additional surface pressure increment recorded. Eucalyptol was allowed to be inhaled by rabbits and lung compliance was monitored. Under our experimental conditions the volatile aromatics exhibited surfactant-like effects, namely a decrease in surface tension between water and air and thus improved lung compliance values in vivo.

Animals↗
PubMed2005

Application of the Taguchi analytical method for optimization of effective parameters of the chemical vapor deposition process controlling the production of nanotubes/nanobeads.

Seven variable parameters of the chemical vapor deposition system have been optimized with the help of the Taguchi analytical method for getting a desired product, e.g., carbon nanotubes or carbon nanobeads. It is observed that almost all selected parameters influence the growth of carbon nanotubes. However, among them, the nature of precursor (racemic, R or Technical grade camphor) and the carrier gas (hydrogen, argon and mixture of argon/hydrogen) seem to be more important parameters affecting the growth of carbon nanotubes. Whereas, for the growth of nanobeads, out of seven parameters, only two, i.e., catalyst (powder of iron, cobalt, and nickel) and temperature (1023 K, 1123 K, and 1273 K), are the most influential parameters. Systematic defects or islands on the substrate surface enhance nucleation of novel carbon materials. Quantitative contributions of process parameters as well as optimum factor levels are obtained by performing analysis of variance (ANOVA) and analysis of mean (ANOM), respectively.

Argon↗
PubMed1985

A comparison of the antibacterial and cytotoxic effects of parachlorophenol.

The antibacterial and cytotoxic effects of parachlorophenol (as 35% camphorated PCP and 2% aqueous PCP) were compared directly, using the agar overlay technique. Human pulp fibroblasts and L929 cells were grown to confluence in 60-mm petri dishes. Bacterial suspensions (S. aureus, E. coli) in agar were poured as a thin layer in 60-mm petri dishes. All were overlaid with agar. Cells were exposed to medicament via filter paper discs placed on the agar surface. After 24 hours, the zone of cell lysis (fibroblasts) or inhibition of bacterial growth was measured as diameter in mm. A dose-response relationship was observed for both fibroblasts and bacteria. The zone of inhibition for fibroblasts was larger than that for bacteria, indicating that the cytotoxicity of parachlorophenol exceeds its antibacterial activity.

Animals↗
PubMed1987

In vitro infection and disinfection of dentinal tubules.

An in vitro model for dentinal tubule infection of root canals was developed. Cylindrical dentin specimens, 4 mm high with a diameter of 6 mm and a canal 2.3 mm wide, were prepared from freshly extracted bovine incisors. The cementum was removed from all dentin blocks. The tubules were opened by four-minute treatments with 17% EDTA and 5.25% NaOCl before being infected with Enterococcus faecalis ATCC 29212 in yeast extract-glucose broth. Bacteria rapidly invaded the tubules. After three weeks of incubation, a heavy infection was found 400 micron from the canal lumen, and the front of the infection reached 1000 micron in some blocks. Camphorated paramonochlorophenol (CMCP) and a calcium hydroxide compound, Calasept, were tested for their disinfecting efficacy toward E. faecalis-infected dentin. Liquid CMCP rapidly and completely disinfected the dentinal tubules, whereas CMCP in gaseous form disinfected tubules less rapidly. Calasept failed to eliminate, even superficially, E. faecalis in the tubules. The method used in bacteriological sampling allowed for sequential removal of 100-micron-thick zones of dentin from the central canal toward the periphery. Control specimens were uniformly infected and yielded growth in bur samples up to some 500 microns from the surface. The model proved quite sensitive and seems suitable for in vitro testing of root canal medicaments.

Animals↗
PubMed1999

In vitro transcriptional analysis of the cytochrome P-450cam hydroxylase operon.

To characterize the promoters of cytochrome P-450cam hydroxylase operon (camDCAB) and the repressor gene (camR), in vitro run-off transcription assays were performed using RNA polymerase (RNAP) holoenzyme reconstituted with the core enzyme and the sigma70 protein of Pseudomonas aeruginosa. Both the mRNAs of camDCAB and camR were accurately transcribed from the respective promoter by the reconstituted RNAP holoenzyme. Both the transcriptions were repressed by CamR protein and the repressions were suppressed by D-camphor, consistent with the regulation in P. putida. These results suggest that the RNA polymerase containing sigma70 recognizes the promoter of camDCAB as well as that of camR.

Bacterial Proteins↗
PubMed2001

In vitro and in vivo estrogenicity of UV screens.

Ultraviolet (UV) screens are increasingly used as a result of growing concern about UV radiation and skin cancer; they are also added to cosmetics and other products for light stability. Recent data on bioaccumulation in wildlife and humans point to a need for in-depth analyses of systemic toxicology, in particular with respect to reproduction and ontogeny. We examined six frequently used UVA and UVB screens for estrogenicity in vitro and in vivo. In MCF-7 breast cancer cells, five out of six chemicals, that is, benzophenone-3 (Bp-3), homosalate (HMS), 4-methyl-benzylidene camphor (4-MBC), octyl-methoxycinnamate (OMC), and octyl-dimethyl-PABA (OD-PABA), increased cell proliferation with median effective concentrations (EC(50)) values between 1.56 and 3.73 microM, whereas butyl-methoxydibenzoylmethane (B-MDM) was inactive. Further evidence for estrogenic activity was the induction of pS2 protein in MCF-7 cells and the blockade of the proliferative effect of 4-MBC by the estrogen antagonist ICI 182,780. In the uterotrophic assay using immature Long-Evans rats that received the chemicals for 4 days in powdered feed, uterine weight was dose-dependently increased by 4-MBC (ED(50 )309mg/kg/day), OMC (ED(50) 935 mg/kg/day), and weakly by Bp-3 (active at 1,525 mg/kg/day). Three compounds were inactive by the oral route in the doses tested. Dermal application of 4-MBC to immature hairless (hr/hr) rats also increased uterine weight at concentrations of 5 and 7.5% in olive oil. Our findings indicate that UV screens should be tested for endocrine activity, in view of possible long-term effects in humans and wildlife.

Animals↗
PubMed2005

Enantiomeric excess determination by fourier transform near-infrared vibrational circular dichroism spectroscopy: simulation of real-time process monitoring.

The first use of near-infrared (NIR) Fourier transform vibrational circular dichroism (FT-VCD) to follow changes in the enantiomeric excess (EE) of chiral sample molecules in time using a flow-cell sampling apparatus is reported. Simultaneous changes in the fractional composition and the EE of a mixture of two different chiral molecules were monitored as a function of time. This simulates the progress of the chemical reaction from a chiral reactant to a chiral product where the mole fractions and EE values of both species may change with time. For the molecules studied, alpha-pinene, camphor, and borneol, the accuracy of following EE changes for one species alone is approximately 2%, while for simultaneously following EE changes in two species it is approximately 3% for 30 min sampling periods at 16 cm(-1) spectral resolution. These findings demonstrate the potential for VCD to be used in the NIR region for real-time monitoring of the composition and %EE of chemical reactions involving the synthesis of chiral molecules.

Bicyclic Monoterpenes↗
PubMedDate not supplied

The role of molecular recognition in charge transport properties of doped polyaniline.

Molecular recognition plays a significant role in the counterion-induced processibility, morphological features, and physical properties of doped polyaniline (PANI). The interaction of the counterion and solvent controls the chain conformation and, as a result, the formation of extended and localized electronic states; hence, it holds the key for tuning a wide range of electrical and optical properties of doped PANI. The combined effects of counterion, solvent, and processing conditions tune the metal-insulator transition, temperature dependence of conductivity, magnetoresistance, and so forth in doped PANI. The typical examples are shown in the case of PANI doped by camphor sulfonic acid, 2-acrylamido-2-methyl-1-propane sulfonic acid, and dodecylbenzoyl sulfonic acid.

Aniline Compounds↗
PubMed1996

Spontaneous comedones on the skin of hairless descendants of Mexican hairless dogs.

In the first experiment, the skin sebum and humidity, perspiration ability of sweat glands, and histology of spontaneous comedones were examined in hairless descendants of Mexican hairless dogs. The skin of females showed lower humidity than that of males. Some animals with a large number of comedones exhibited remarkably high skin sebum scores. The comedones were distributed throughout the dorsal skin, and a cluster of lesions was found mainly in the limbs and prepuces. The sweat glands showed no perspiration in the sudorific test. Histologically, both infant and adult animals had lesions of micro- and/or "blackhead" comedones. Plugged follicles containing abundant keratic substances associated well-developed sebaceous glands. Spontaneous comedones in the skin of hairless dogs were grossly and histologically similar to the acne vulgaris observed in human beings. The skin of some adult animals showed a large number of protrusive comedones which were solid cystic structures containing organized substances. In the second experiment, three kinds of antiacne agents (sulfur and camphor, sulfur and resorcinol, and ibuprofen piconol) were applied daily to the test sites for one month. These antiacne agents caused prominent extrusion of keratin plugs from follicular sites. The results suggest that the hairless dogs are a predictive model for evaluating the efficacy of antiacne agents proposed for acne treatment.

Acne Vulgaris↗