Characterization of Jamaican agro-industrial wastes. Part II, fatty acid profiling using HPLC: precolumn derivatization with phenacyl bromide.

This paper describes the determination of fatty acid composition of coffee, citrus and rum distillery wastes using reversed-phase high-performance liquid chromatography (RP-HPLC). Lipid extracts of the waste samples are derivatized with phenacyl bromide and their phenacyl esters are separated on a C8 reversed-phase column by using continuous gradient elution with water and acetonitrile. The presence of saturated and unsaturated fatty acids in quantifiable amounts in the examined wastes, as well as the high percentage recoveries, are clear indications that these wastes have potential value as inexpensive sources of lipids. The HPLC procedures described here could be adopted for further analysis of materials of this nature.

Biodegradation of bisphenol-A in river sediment.

This research investigated the aerobic and anaerobic degradation of bisphenol-A (BPA) in river sediment. With the addition of 250 µg g⁻¹ BPA, the percentages of BPA remaining in sediment from sites A, B, and C were 21.9 %, 3.5 % and 12.5 %, respectively, after 5 days of incubation under aerobic conditions; degradation was not significant after 140 days of incubation under anaerobic conditions. The aerobic degradation of BPA was enhanced by adding yeast extract (5 mg L⁻¹), sodium chloride (1 %), cellulose (0.96 mg L⁻¹), brij 30 (55 µM), brij 35 (91 µM), rhamnolipid (130 mg L⁻¹), or surfactin (43 mg L⁻¹), with rhamnolipid yielding higher BPA degradation than the other additives. 2,4-bis (1,1-dimethyl ethyl) phenol, an intermediate product resulting from the aerobic degradation of BPA was accumulated in sediments. Of the bacterial strains isolated from the sediment, strains J1, J2, J3, and J4 expressed the best aerobic degrading ability. The highest BPA degradation rate was found in the sediment by the addition of strains J1, J2, J3, and J4 combined, whereas the sediment without the addition of the 4 strains had the lowest biodegradation rate. This research offers feasible methods for the removal of BPA in river sediment for bioremediation.

Biodegradation of phthalate esters in polluted soil by using organic amendment.

This study investigated the biodegradation of the phthalate esters (PAEs) di-n-butyl phthalate (DBP) and di-(2-ethyl hexyl) phthalate (DEHP) in sludge and sludge-amended soil. DBP (100 mg kg(-1)) and DEHP (100 mg kg(-1)) were added to sewage sludge, which was subsequently added to soil. The results showed that sewage sludge can degrade PAEs and the addition of sewage sludge to soil enhanced PAE degradation. Sludge samples were separated into fractions with various particle size ranges, which spanned 0.1-0.45 µm to 500-2000 µm. The sludge fractions with smaller particle sizes demonstrated higher PAE degradation rates. However, when the different sludge fractions were added to soil, particle size had no significant effect on the rate of PAE degradation. The results from this study showed that microbial strains F4 (Rhodococcus sp.) and F8 (Microbacterium sp.) were constantly dominant in the mixtures of soil and sludge.

Biodegradation of tetrachlorobisphenol-A in river sediment and the microbial community changes.

This study investigated the aerobic degradation of tetrachlorobisphenol-A (TCBPA) in sediment samples collected at three sites along Erren River in southern Taiwan. TCBPA biodegradation rate constants (k(1)) and half-lives (t(1/2)) ranged from 0.03 to 0.06 day(-1) and 11.6 to 23.1 days, respectively. The biodegradation of TCBPA was enhanced by the addition of cellulose, yeast extract, sodium chloride, brij 30, brij 35, surfactin, and rhamnolipid. Of the micro-organism strains isolated from the sediment, we found that strains L1 and L5 (identified as Bacillus megaterium and Pseudomonas putida) expressed the best biodegrading ability. The inoculation of sediment with the TCBPA-degrading bacteria could enhance the efficiency of degradation.

Characterization of Jamaican agro-industrial wastes. Part I: characterization of amino acids using HPLC: pre-column derivatization with phenylisothiocyanate.

Jamaican agro-industries generate large quantities of wastes, which are either discarded or under-utilized. In order to evaluate the possible utilization of these wastes, it is necessary that the profiles of the major biochemical groups be developed. This paper describes the determination of the amino acid composition of coffee, citrus, and rum distillery wastes using a reversed-phase high-performance liquid chromatography method. Acid hydrolysates of the wastes are derivatized with phenylisothiocyanate. They are analyzed as their phenylthiocarbamyl derivatives and determined quantitatively using norleucine as the internal standard. The presence of all the 17 amino acids investigated, nine of which include those essential for animal nutrition, are observed in the samples investigated, suggesting a high quality of protein with implications in the formulation of animal feeds.

Biodegradation of phenanthrene and pyrene in compost-amended soil.

This study investigated the biodegradation of the polycyclic aromatic hydrocarbons (PAHs) phenanthrene and pyrene in compost and compost-amended soil. The degradation rates of the two PAHs were phenanthrene>pyrene. The degradation of PAH was enhanced when the two PAHs were present simultaneously in the soil. The addition of either of the two types of compost (straw and animal manure) individually enhanced PAH degradation. Compost samples were separated into fractions with various particle size ranges, which spanned 2-50 microm, 50-105 microm, 105-500 microm, and 500-2000 microm. We observed that the compost fractions with smaller particle sizes demonstrated higher PAH degradation rates. However, when the different compost fractions were added to soil, compost particle size had no significant effect on the rate of PAH degradation. Of the micro-organisms isolated from the soil-compost mixtures, strains S1, S2, and S8, which were identified as Arthrobacter nicotianae, Pseudomonas fluorescens, and Bordetella Petrii, respectively, demonstrated the best degradation ability.

Biodegradation of phenanthrene and pyrene from mangrove sediment in subtropical Taiwan.

This study investigated the effects of various factors on the biodegradation of the polycyclic aromatic hydrocarbons (PAHs) phenanthrene and pyrene, and characterized the structure of the microbial community in mangrove sediment from subtropical Taiwan. The degradation of PAH was enhanced by the addition of brij 30, tween 80, yeast extract, hydrogen peroxide, sodium chloride or cellulose. However, PAH degradation was inhibited by the addition of humic acid, di-(2-ethylhexyl), phthalate (DEHP) or nonylphenol. Our results also showed that the addition of various substrates changed the microbial community in mangrove sediment. The strains MS2 (Dyella ginsengisoli) and MS4 (Bacillus pumilus) were the constantly dominant bacteria under various treatments in the mangrove sediment.

Immunological evidence supporting the use of extracts from Boehmeria jamaicensis Urb for treating the common cold and sinus infections

Mixed lymphocyte responses assays were conducted at 25.0 and 250.0 microg/mL of the crude ethanolic extract of Boehmeria jamaicensis Urb (coded as BJE) using peripheral lymphocytes obtained from individuals suffering from the common cold after four days of infection and from healthy individuals (without the common cold infection). At a concentration of 25 ug/mL, gamma interferon (IFN-gamma) was increased by 24.03 fold and interleukin 4 (IL-4) by 1.71 fold for the cells obtained from individuals with the common cold (Group A). The extract suppressed IFN-gamma by 8.3% while IL-4 was stimulated by 9.90 fold from peripheral lymphocytes obtained from healthy individuals (Group B). Gamma interferon was suppressed at 250 microg/mL while IL-4 was elevated by 1.86 fold for cells obtained from individuals suffering from the common cold (Group A). In conclusion, BJE could have implications for the treatment of the common cold.

Ensayos de reacci¨®n linfocitaria mixta fueron realizados a 25.0 y 250.0 ¦Ìg/mL de extracto etan¨®lico crudo de Boehmeria jamaicensis Urb (codificado como BJE), usando linfocitos perif¨¦ricos obtenidos de individuos con catarro com¨²n luego de cuatro d¨ªas de infecci¨®n, y de individuos sanos (sin la infecci¨®n del catarro com¨²n). Se hall¨® que el interfer¨®n-gamma (IFN-¦Ã) aument¨® en 24.03 veces, y la interleucina 4 (IL-4) en 1.71 veces para las c¨¦lulas obtenidas de individuos con catarro com¨²n, a 25¦Ìg/mL. El extracto inhibi¨® IFN-¦Ã en un 8.3 % en tanto que el IL-4 fue estimulado en 9.90 veces a partirde los linfocitos perif¨¦ricos obtenidos de individuos sanos. El gamma-interfer¨®n fue inhibido a 250 ¦Ìg/mL, mientras que la IL-4 se elev¨® en 1.86 veces para las c¨¦lulas obtenidas de individuos que sufren de catarro com¨²n.

Anaerobic degradation of nonylphenol in soil.

This study investigated the effects of various factors on the anaerobic degradation of nonylphenol (NP) in soil. The results show that the optimal pH for NP degradation was 7.0 and that the degradation rate was enhanced when the temperature was increased. The addition of compost enhanced NP degradation. The individual addition of the electron donors lactate, acetate, and pyruvate inhibited NP degradation. The high-to-low order of NP degradation rates under three anaerobic conditions was sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The results show that sulfate-reducing bacteria, methanogen, and eubacteria are involved in the anaerobic degradation of NP, with sulfate-reducing bacteria being a major component of the soil. Of the anaerobic strains isolated from the soil samples, strain AT3 expressed the best ability to biodegrade NP.

Anaerobic degradation of five polycyclic aromatic hydrocarbons from river sediment in Taiwan.

This study investigated the anaerobic degradation of five polycyclic aromatic hydrocarbons (PAHs) from Erren River sediment in southern Taiwan. The degradation rates of PAH were in the order: acenaphthene > fluorene > phenanthrene > anthracene > pyrene. The degradation rate was enhanced when the five compounds were present simultaneously in river sediment. Comparison of the PAH degradation rates under three reducing conditions showed the following order: sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The addition of electron donors (acetate, lactate and pyruvate) enhanced PAH degradation under methanogenic and sulfate-reducing conditions. However, the addition of acetate, lactate or pyruvate inhibited PAH degradation under nitrate-reducing conditions. The addition of heavy metals, nonylphenol and phthalate esters (PAEs) inhibited PAH degradation. Our results show that sulfate-reducing bacteria, methanogen and eubacteria are involved in the degradation of PAH; sulfate-reducing bacteria constitute a major microbial component in PAH degradation. Of the microorganism strains isolated from the sediment samples, we found that strain ER9 expressed the greatest biodegrading ability.

Immunological evidence supporting the use of extracts from Boehmeria jamaicensis Urb for treating the common cold and sinus infections.

Mixed lymphocyte responses assays were conducted at 25.0 and 250.0 microg/mL of the crude ethanolic extract of Boehmeria jamaicensis Urb (coded as BJE) using peripheral lymphocytes obtained from individuals suffering from the common cold after four days of infection and from healthy individuals (without the common cold infection). At a concentration of 25 ug/mL, gamma interferon (IFN-gamma) was increased by 24.03 fold and interleukin 4 (IL-4) by 1.71 fold for the cells obtained from individuals with the common cold (Group A). The extract suppressed IFN-gamma by 8.3% while IL-4 was stimulated by 9.90 fold from peripheral lymphocytes obtained from healthy individuals (Group B). Gamma interferon was suppressed at 250 microg/mL while IL-4 was elevated by 1.86 fold for cells obtained from individuals suffering from the common cold (Group A). In conclusion, BJE could have implications for the treatment of the common cold.

High-performance liquid chromatographic analysis of amino acids in ackee fruit with emphasis on the toxic amino acid hypoglycin A.

High-performance liquid chromatography is used to determine the amino acid content of ripe and unripe ackee fruit. Specific emphasis is placed on the level of the toxic amino acid hypoglycin A (hyp-A) in the unripe and ripe ackee fruit and seed. Unripe samples are found to contain significantly higher quantities (P < 0.05) of hyp-A when compared with ripe samples. Uncooked unripe fruit is found to contain 124.4 +/- 6.7 mg/100 g fresh weight and uncooked ripe fruit 6.4 +/- 1.1 mg/100 g fresh weight. The seed of the uncooked unripe fruit is found to contain 142.8 +/- 8.8 mg/100 g fresh weight, and the seed of uncooked ripe fruit has 106.0 +/- 5.4 mg/100 g fresh weight. Boiling fruit in water for approximately 30 min is efficient in removing hyp-A from the edible arilli; however, low levels of 0.54 +/- 0.15 mg/200 mL are detected in the water that was used to cook the ripe fruit. The average %recovery of the amino acids was 80.34%.

Arginine-modulated receptor-activated calcium influx via a NO/cyclic GMP pathway in human SK-N-SH neuroblastoma cells.

The effects of arginine on calcium mobilization in human SK-N-SH neuroblastoma cells were examined. It was found that arginine potentiated an increase in carbachol-induced Ca2+ from the external Ca2+ influx as opposed to an internal Ca2+ release from intracellular pools. The potentiation effect of arginine on carbachol-induced calcium mobilization was mimicked by either 8-bromo cyclic GMP or sodium nitroprusside. In addition, it was found that arginine induced NO production and an increase in cyclic GMP. Moreover, arginine-induced potentiation, NO production, and cyclic GMP increases were all suppressed after the preincubation of cells with N-methyl-L-arginine or N-nitro-L-arginine, nitric oxide synthase inhibitor. It is suggested that the NO production and subsequent cyclic GMP elevation induced by arginine are responsible for the potentiation of carbachol-induced Ca2+ increase. Our results show the existence of a NO/cyclic GMP pathway and an interconnection of NO and Ca2+ signaling pathways in human SK-N-SH neuroblastoma cells. We also observed that NO, which is produced by endothelial CPAE cells, has a modulating effect on cyclic GMP elevation in human SK-N-SH neuroblastoma cells. The intercellular communication role of NO and its cell-diffusing character may also affect the regulation of nonneuronal cells in their interactions with neuronal cells.

Streptococci and Pasteurella spp. associated with disease of the equine lower respiratory tract.

The likelihood of finding evidence of inflammation in 551 tracheal washes collected endoscopically from 278 Thoroughbred racehorses increased with the number of bacterial colony forming units (cfu) per ml of wash (P < 0.001). The aerobic bacteria Streptococcus zooepidemicus, Pasteurella/Actinobacillus-like species and Streptococcus pneumoniae were significantly associated with lower airway inflammation whereas coagulase-negative Staphylococcus spp., alpha-haemolytic Streptococcus spp., Acinetobacter spp., Bacillus spp., Escherichia coli, Pseudomonas aeruginosa, non-haemolytic Streptococcus spp. and Enterobacteriaceae were not; Bordetella bronchiseptica was not isolated. Lower airway inflammation was particularly associated with bacteria in horses < or = 3 years of age. S. zooepidemicus, S. pneumoniae and Pasteurella/Actinobacillus-like species were isolated from 167 of 551 washes, either alone or in combination.

Partial reversion of transformed phenotype of B104 cancer cells by antisense nucleic acids.

We used antisense RNA to inhibit the expression of oncogene neu and investigated the effects of diminished neu expression on the phenotypes of B104 cells containing activated oncogene neu. Antisense MT-neu and pSV-neo plasmids were cotransfected into neuroblastoma B104 cells. Southern analysis showed the integration of anti-neu DNA into B104 cells. The expression of neu was inhibited up to 90% as quantitated by immunoprecipitation. The growth rate and the potential to differentiate in these transfectants were not affected as compared to the parental cell lines. The ability to grow in soft agar was inhibited more than 90% in these transfectants. Our results indicated that antisense-RNA against a specific oncogene can decrease the tumorigenicity of tumor cells but may not be able to revert it to normal cells completely.

Release of the p53-induced repression on thymidine kinase promoter by single p53-binding sequence.

Herpes simplex thymidine kinase (TK) promoter was shown to be repressed by the wild-type p53. Using a model system that the p53-binding site was linked to the thymidine kinase promoter, we demonstrated that single p53-specific binding site was sufficient to abolish the repression. On the contrary, the mutant p53 had the opposite effects on the HSV-TK promoter in BHK cells. The results suggest that the p53-binding site may act as an enhancer to regulate the gene expression in a novel way in vivo.

The effects of glucocorticoid hormone on the expression of c-jun.

The effects of glucocorticoid hormone on the expression of c-jun in the fibroblasts were studied. The expression of c-jun was repressed by dexamethasone in the NIH3T3 cells, but not in the transformed B104-1 or EJ-Ras cells. The repression was not relieved by the addition of cycloheximide.

Interaction of acridines and tetrahydroacridines with DNA at low DNA/dye ratios.

This study examines certain aspects of the interaction of acridines with DNA. A comparative study of the methods available for the determination of the association constants (Kap) for compounds which interact with DNA has been pursued. A new equation which permits the spectrophotomeric determination of Kap has been derived. This equation can be applied to compounds which upon interaction with a polymer exhibit discrete absorption changes. Application of this equation to substituted acridines and tetrahydroacridines yields some preliminary information on the effect of ring substituents on the interaction of acridines with DNA. Low levels of DNA/dye ratios have been used in the studies reported herein.

Diacridines: bifunctional intercalators. I. Chemistry, physical chemistry and growth inhibitory properties.

The synthesis, as well as the rationale for synthesis of diacridines, double intercalators, as potential inhibitors of nucleic acid synthesis is presented. The syntheses of (9-acridyl)-putrescine and -spermine, and bis(-9-acridyl)-putrescine, -spermidine, -spermine diamines and of bis(6-chloro-2-methoxy-9-acridyl)-putrescine and -spermine diamines, all substituted on the terminal NH2 groups are described. In addition, the homologous series of diacridines connected by the amino groups of the diamines NH2(CH2)nNH2 (where n = 2,3,4,6,8,10,12,14,16,18) to the C-9 of the diacridines has been synthesized. The chemical properties of these compounds as well as their molecular relationship to DNA are presented. The effect of the double intercalators on the Tm of DNA and of (A)n - (U)n, (dA)n - (dT)n, (G)n - (C)n and on (dG)n - (dC)n have been determined. The double acridine intercalators produce a much greater increase of the Tm of these nucleic acids than do the single acridine intercalators. They also profoundly affect the Tm of DNA in physiological salt concentrations; under these latter conditions the single intercalators have no effect. The relationship between the length of the chain connecting the two acridine rings and the inhibition of the growth of P-388 cells in vitro and vivo is presented. Their growth inhibitory properties appear, in general, to parallel their intercalative abilities.