Antimicrobial effects of herbal extracts on Streptococcus mutans and normal oral streptococci

Streptococcus mutans is associated with dental caries. A cariogenic biofilm, in particular, has been studied extensively for its role in the formation of dental caries. Herbal extracts such as Cudrania tricuspidata, Sophora flavescens, Ginkgo biloba, and Betula Schmidtii have been used as a folk remedy for treating diseases. The purpose of this study was to evaluate and compare the antibacterial activity of herbal extracts against normal oral streptococci, planktonic and biofilm of S. mutans. Streptococcus gordonii, Streptococcus oralis, Streptococcus salivarius, Streptococcus sanguinis, and S. mutans were cultivated with brain heart infusion broth and susceptibility assay for the herbal extracts was performed according to the protocol of Clinical and Laboratory Standard Institute. Also, S. mutans biofilm was formed on a polystyrene 12-well plate and 8-well chamber glass slip using BHI broth containing 2% sucrose and 1% mannose after conditioning the plate and the glass slip with unstimulated saliva. The biofilm was treated with the herbal extracts in various concentrations and inoculated on Mitis-Salivarius bacitracin agar plate for enumeration of viable S. mutans by counting colony forming units. Planktonic S. mutans showed susceptibility to all of the extracts and S. mutans biofilm exhibited the highest level of sensitivity for the extracts of S. flavescens. The normal oral streptococci exhibited a weak susceptibility in comparison to S. mutans. S. oralis, however, was resistant to all of the extracts. In conclusion, the extract of S. flavescens may be a potential candidate for prevention and management of dental caries.     

Abscisic Aldehyde Is an Intermediate in the Enzymatic Conversion of Xanthoxin to Abscisic Acid in Phaseolus vulgaris L. Leaves

The enzymatic conversion of xanthoxin to abscisic acid by cell-free extracts of Phaseolus vulgaris L. leaves has been found to be a two-step reaction catalyzed by two different enzymes. Xanthoxin was first converted to abscisic aldehyde followed by conversion of the latter to abscisic acid. The enzyme activity catalyzing the synthesis of abscisic aldehyde from xanthoxin (xanthoxin oxidase) was present in cell-free leaf extracts from both wild type and the abscisic acid-deficient molybdopterin cofactor mutant, Az34 (nar2a) of Hordeum vulgare L. However, the enzyme activity catalyzing the synthesis of abscisic acid from abscisic aldehyde (abscisic aldehyde oxidase) was present only in extracts of the wild type and no activity could be detected in either turgid or water stressed leaf extracts of the Az34 mutant. Furthermore, the wilty tomato mutants, sitiens and flacca, which do not accumulate abscisic acid in response to water stress, have been shown to lack abscisic aldehyde oxidase activity. When this enzyme fraction was isolated from leaf extracts of P. vulgaris L. and added to extracts prepared from sitiens and flacca, xanthoxin was converted to abscisic acid. Abscisic aldehyde oxidase has been purified about 145-fold from P. vulgaris L. leaves. It exhibited optimum catalytic activity at pH 7.25 in potassium phosphate buffer.     

Protein synthesis in chloroplasts: V. Translation of messenger RNA for the large subunit of fraction I protein in a heterologous cell-free system

The addition of spinach chloroplast total RNA to cell-free extracts from Escherichia coli stimulates amino acid incorporation into protein. The products were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and were qualitatively and quantitatively similar to those synthesized in intact isolated chloroplasts. There are two major discrete products of both systems with molecular weights of 52,000 and 35,000. The [³⁵S]methionine-containing chymotryptic peptides of the 52,000 M_r polypeptide synthesized in the E. coli cell-free system have been compared with those of fraction I protein large subunit labelled with [³⁵S]methionine in vivo. From the close similarity in chromatographic properties of the peptides of the two polypeptides, we conclude that the 52,000 M_r product of chloroplast RNA-directed protein synthesis in E. coli extracts is the large subunit of fraction I protein.     

Structural studies of acidic glucomannans from strains of Serratia marcescens O14 and O6

Partially acetylated acidic glucomannans have been isolated from three strains of Serratia marcescens serogroup O14 and one strain of the cross-reacting serogroup O6. Degradative and spectroscopic studies established that the polysaccharides have branched tetrasaccharide repeating-units of the structure shown. Individual polymers may vary in the extent or location of O-acetylation, and in the extent of undefined heterogeneity apparently associated with the glucosyluronic acid residues. Although the polymers were obtained from lipopolysaccharide extracts, there are indications of a (micro)capsular origin. The acidic glucomannans may constitute a common antigen which defines the O14-O6 complex of S. marcescens.     

Searsia species with affinity to the N-methyl-d-aspartic acid (NMDA) receptor

Species of Searsia are used in traditional medicine to treat epilepsy. Previous studies on S. dentata and S. pyroides have shown that this is likely mediated via the N-methyl-d-aspartic acid (NMDA) receptor. Ethanolic extracts of leaves of six Searsia species were tested in a binding assay for affinity to the NMDA-receptor. S. dentata, S. gerrardii, S. glauca, S. natalensis, S. pentheri and S. pyroides all contained compounds with affinity to the NMDA-receptor. The species with the best affinity was S. pentheri. This might make it possible to use the Searsia species interchangeably, so the more accessible Searsia species can be used in traditional medicine.     

Fermentation of Aqueous Plant Seed Extracts by Lactic Acid Bacteria

The effects of lactic acid bacterial fermentation on chemical and physical changes in aqueous extracts of cowpea (Vigna unguiculata), peanut (Arachis hypogea), soybean (Glycine max), and sorghum (Sorghum vulgare) were studied. The bacteria investigated were Lactobacillus helveticus, L. delbrueckii, L. casei, L. bulgaricus, L. acidophilus, and Streptococcus thermophilus. Organisms were inoculated individually into all of the seed extracts; L. bulgaricus and S. thermophilus were also evaluated together as inocula for fermenting the legume extracts. During fermentation, bacterial population and changes in titratable acidity, pH, viscosity, and color were measured over a 72-h period at 37°C. Maximum bacterial populations, titratable acidity, pH, and viscosity varied depending upon the type of extract and bacterial strain. The maximum population of each organism was influenced by fermentable carbohydrates, which, in turn, influenced acid production and change in pH. Change in viscosity was correlated with the amount of protein and titratable acidity of products. Color was affected by pasteurization treatment and fermentation as well as the source of extract. In the extracts inoculated simultaneously with L. bulgaricus and S. thermophilus, a synergistic effect resulted in increased bacterial populations, titratable acidity, and viscosity, and decreased pH in all the legume extracts when compared to the extracts fermented with either of these organisms individually. Fermented extracts offer potential as substitutes for cultured dairy products.     

The effects of the piroplasm Babesia bigemina on the survival and reproduction of the blue tick,Boophilus decoloratus

Seven strains of the insect pathogenic fungus Verticillium lecanii have been examined in the Calliphora erythrocephala bioassay for the production, in surface culture on Czapek Dox medium, of insecticidal secondary metabolites. One strain which had lost its pathogenicity on storage yielded no active compounds. The remainder yielded dipicolinic acid (pyridine-2,6-dicarboxylic acid) which was responsible for the insecticidal activity of acidic extracts. Neutral extracts from two strains contained novel insecticidal C₂₅ compounds.     

Mycopesticidal effects of characterized extracts of Penicillium isolates and purified secondary metabolites (including mycotoxins) on Drosophila melanogaster and Spodoptora littoralis

The characterized crude extracts of seven Penicillium isolates were bioassayed against Spodoptora littoralis larvae for weight reduction and mortality. A total of 15 metabolites were detected in the extracts, 13 of which were named. Five extracts caused significant reductions in weight and four of these caused significant increases in mortality, both at the P < 0.05 level. Seven purified secondary metabolites selected from those present in the crude extracts were bioassayed at 10 ppm against Drosophila melanogaster for feeding inhibition and S. littoralis for feeding inhibition and mortality. Significant effects (P < 0.05) were observed in all three assays for ochratoxin A, brevianamide A, citrinin, and penicillic acid. Viomellein significantly (P < 0.05) reduced feeding in D. melanogaster and survival in S. littoralis. Cyclopenol significantly (P < 0.05) inhibited feeding in both insects but not survival of S. littorolis. Significant effects were not observed for patulin (P > 0.05). High levels of feeding inhibition were obtained for brevianamide A (84.4 ± 18.21%) and penicillic acid (95.1 ± 1.21 %) against S. littoralis. Cyclopenol and brevianamide A were found to be antagonistic, and this is the first valid toxicity data ascribed to these metabolites. The implications of these results with respect to potential mycopesticides are considered.     

Bioefficacy of some Rhizobactrial isolates against sorghum root Rot pathogen Bipolaris sorokiniana

This study carried out to identify certain microbial allelochemicals with antifungal activity of some rhziobacterial isolates against Bipolaris sorokiniana fungi. The fungicidal activity of isolated microbe metabolites was compared based on inhibition % of fungal growth. Results showed that ethyl acetate crude extracts with two concentrations (500 and 1000 ppm) of Pseudomonas geniculata (SC) and Bacillus cereus (S4) were the most efficient isolates recorded inhibition % 33.62 and 52.59% followed by S4 (Bacillus cereus (ATCC 14579) which achieved inhibition % 33.62 and 46.55% at the same concentrations, respectively. After 4 days.The constituents analyzed by LC-MS/MS and FTIR of the ethyl acetate extracts of the Pseudomonas geniculata ATCC19374 were afforded aminobutyric acid, 1,4-benzoquinone, coumaric acid, sinapic acid, tryptophan amino acid, Succinic acid and ferulic acid. While, the secondary metabolites of (Bacillus cereus ATCC 14579 extract were aminobutyric acid, 1,4-benzoquinone, coumaric acids, sinapic acid, ferulic acid and benzoic acid. Results indicated that the isolates of Pseudomonas geniculata ATCC19374 and Bacillus cereus ATCC 14579 could be use as a good element in plant root rot pathogen Bipolaris sorokiniana management.     

Saccharopine and 2-aminoadipic acid in Reseda odorata

2(S),2′(S)-N⁶-(2′-Glutaryl)lysine (l-saccharopine) and 2(S)-2-aminoadipic acid have been isolated from Reseda odorata. When traditional isolation procedures are used l-pyrosaccharopine (5(S),5′(S)-N-(5′-amino-5′-carboxy-pentyl)-2-pyrrolidone-5-carboxylic acid) is formed from l-saccharopine by lactamisation. The degree of lactamisation during various isolation steps has been studied, The amino acids were identified by IR and PMR spectroscopy and the configurations established by enzymic and polarimetric analyses. The contents of saccharopine and 2-amino-adipic acid have been determined relative to the total nitrogen content at various stages in the growth cycle of R. odorata.     

Anti-inflammatory,antioxidant, anti-cholinesterase activity and mutagenicity of South African medicinal orchids

While the role of various processes in inflammatory-related degenerative disorders is still being researched, many avenues of research have concentrated on the treatment and/or prevention of these disorders. Inflammatory-responses, the cholinergic system and oxidative stress have often been linked to the symptoms prevalent in aged persons and Alzheimer's patients. The current research explored the selective inhibition of cyclooxygenase (COX) enzymes, antioxidant and anti-cholinesterase activities of selected South African orchid extracts, currently traded in herbal markets along the east coast of South Africa. Out of a total of 53 evaluated extracts, significant anti-inflammatory activity was observed in nearly 40% of extracts in the COX-1 assay and 25% of extracts in the COX-2 assay. Overall, the DCM root extract of Ansellia africana was the most potent, the DCM tuber extract of Eulophia hereroensis was the only extract to significantly inhibit both COX enzymes, while all Bulbophyllum scaberulum organic root extracts exhibited COX-2 selective inhibitory activity. Bulbophyllum scaberulum DCM root extract was also the most effective anti-cholinesterase extract, performing better than galanthamine. In the single electron transfer (SET) (2,2′-diphenylpicrylhydrazyl (DPPH) free radical scavenging assay and ferric reducing antioxidant power (FRAP) reaction based assays, E. petersii pseudobulb and A. africana root extracts performed better than other extracts. In the hydrogen atom transfer (HAT) (β-carotene/linoleic acid assay) reaction-based assay, the leaf extract of Tridactyle tridentata and root extracts of Cyrtorchis arcuata and E. hereroensis exhibited the best antioxidant effects. The mutagenicity (Salmonella/microsome assay) was also determined. Organic leaf and root extracts of C. arcuata produced no genotoxic effects in comparison to the other tested species. None of the crude extracts tested demonstrated mutagenic effects using S. typhimurium strain TA98 with metabolic activation. The results obtained in this study validate the use of certain orchid species in South African traditional medicine for inflammation-related degenerative disorders.     

Antimicrobial Activity of Antrodia camphorata Extracts against Oral Bacteria

Antrodia camphorata (A. camphorata) is a unique, endemic and extremely rare mushroom species native to Taiwan, and both crude extracts of and purified chemical compounds from A. camphorata have been reported to have a variety of significant beneficial effects, such as anti-tumor and anti-inflammatory activity. However, reports on the effects of A. camphorata against dental pathogens have been limited. Oral health is now recognized as important for overall general health, including conditions such as dental caries, periodontal disease and rheumatoid arthritis. Streptococcus mutans (S. mutans) and Porphyromonas gingivalis (P. gingivalis) are the most common bacteria associated with dental plaque and periodontopathic diseases, respectively. Thus, our study examined the ability of five various crude extracts of A. camphorata to inhibit the growth of dental bacteria and anti-adherence in vitro. Among the extracts, the ethanol, ethyl acetate and chloroform extracts exhibited the lowest MICs against P. gingivalis and S. mutans (MIC = 4∼16 µg/mL). The MIC of the aqueous extract was greater than 2048 µg/mL against both P. gingivalis and S. mutans. In vitro adherence of S. mutans was significantly inhibited by the addition of either the ethyl acetate extract or chloroform extract (MIC = 16∼24 µg/mL), while the ethanol extract (MIC = 32∼64 µg/mL) exhibited moderate inhibitory activity. Based on the result of this study, the ethyl acetate and chloroform extracts of A. camphorata may be good candidates for oral hygiene agents to control dental caries and periodontopathic conditions.     

Intracellular α-Amylase of Streptococcus mutans

Sequencing upstream of the Streptococcus mutans gene for a CcpA gene homolog, regM, revealed an open reading frame, named amy, with homology to genes encoding α-amylases. The deduced amino acid sequence showed a strong similarity (60% amino acid identity) to the intracellular α-amylase of Streptococcus bovis and, in common with this enzyme, lacked a signal sequence. Amylase activity was found only in S. mutans cell extracts, with no activity detected in culture supernatants. Inactivation of amy by insertion of an antibiotic resistance marker confirmed that S. mutans has a single α-amylase activity. The amylase activity was induced by maltose but not by starch, and no acid was produced from starch. S. mutans can, however, transport limit dextrins and maltooligosaccharides generated by salivary amylase, but inactivation of amy did not affect growth on these substrates or acid production. The amylase digested the glycogen-like intracellular polysaccharide (IPS) purified from S. mutans, but the amy mutant was able to digest and produce acid from IPS; thus, amylase does not appear to be essential for IPS breakdown. However, when grown on excess maltose, the amy mutant produced nearly threefold the amount of IPS produced by the parent strain. The role of Amy has not been established, but Amy appears to be important in the accumulation of IPS in S. mutans grown on maltose.     

Metabolic regulation of apoB mRNA editing is associated with phosphorylation of APOBEC-1 complementation factor

Apolipoprotein B (apoB) mRNA editing is a nuclear event that minimally requires the RNA substrate, APOBEC-1 and APOBEC-1 Complementation Factor (ACF). The co-localization of these macro-molecules within the nucleus and the modulation of hepatic apoB mRNA editing activity have been described following a variety of metabolic perturbations, but the mechanism that regulates editosome assembly is unknown. APOBEC-1 was effectively co-immunoprecipitated with ACF from nuclear, but not cytoplasmic extracts. Moreover, alkaline phosphatase treatment of nuclear extracts reduced the amount of APOBEC-1 co-immunoprecipitated with ACF and inhibited in vitro editing activity. Ethanol stimulated apoB mRNA editing was associated with a 2- to 3-fold increase in ACF phosphorylation relative to that in control primary hepatocytes. Significantly, phosphorylated ACF was restricted to nuclear extracts where it co-sedimented with 27S editing competent complexes. Two-dimensional phosphoamino acid analysis of ACF immunopurified from hepatocyte nuclear extracts demonstrated phosphorylation of serine residues that was increased by ethanol treatment. Inhibition of protein phosphatase I, but not PPIIA or IIB, stimulated apoB mRNA editing activity coincident with enhanced ACF phosphorylation in vivo. These data demonstrate that ACF is a metabolically regulated phosphoprotein and suggest that this post-translational modification increases hepatic apoB mRNA editing activity by enhancing ACF nuclear localization/retention, facilitating the interaction of ACF with APOBEC-1 and thereby increasing the probability of editosome assembly and activity.     

Antimicrobial activity of some plant extracts against bacterial strains causing food poisoning diseases

Prevention of food spoilage and food poisoning pathogens is usually achieved by use of chemical preservatives which have negative impacts including: human health hazards of the chemical applications, chemical residues in food & feed chains and acquisition of microbial resistance to the used chemicals. Because of such concerns, the necessity to find a potentially effective, healthy safer and natural alternative preservatives is increased. Within these texts, Plant extracts have been used to control food poisoning diseases and preserve foodstuff. Antimicrobial activity of five plant extracts were investigated against Bacillus cereus, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Salmonella typhi using agar disc diffusion technique. Ethanolic extracts of Punica granatum, Syzygium aromaticum, Zingiber officinales and Thymus vulgaris were potentially effective with variable efficiency against the tested bacterial strains at concentration of 10 mg/ml while extract of Cuminum cyminum was only effective against S. aureus respectively. P. granatum and S. aromaticum ethanolic extracts were the most effective plant extracts and showed bacteriostatic and bactericidal activities against the highly susceptible strains of food borne pathogenic bacteria (S. aureus and P. aeruginosa) with MIC's ranged from 2.5 to 5.0 mg/ml and MBC of 5.0 and 10 mg/ml except P. aeruginosa which was less sensitive and its MBC reached to 12.5 mg/ml of S. aromaticum respectively. These plant extracts which proved to be potentially effective can be used as natural alternative preventives to control food poisoning diseases and preserve food stuff avoiding healthy hazards of chemically antimicrobial agent applications.     

Diplodiatoxin,chaetoglobosins, and diplonine associated with a field outbreak of Stenocarpella ear rot in Illinois

Stenocarpella maydis causes a fungal dry-rot of maize ears and is associated with diplodiosis, a neuromycotoxicosis in cattle grazing harvested maize fields in southern Africa and Argentina. There have been no reports of Stenocarpella metabolites in maize crop residues. Chemical investigations of S. maydis-infected grain from ears exhibiting different levels of ear rot severity following a 2010 field outbreak of Stenocarpella ear rot in Illinois led to the detection of diplodiatoxin and chaetoglobosins M and O as major components in the ethyl acetate extracts by LC-MS. Following post-harvest moist incubation of the S. maydis-infected grain, the amounts of each compound increased (approx. tenfold) and chaetoglobosin K was detected as a dominant toxin. In separate ¹H NMR-based analyses, the neurotoxin diplonine was detected as a minor component in methanol extracts of S. maydis-infected grain as well as cultures of S. maydis isolates from Midwest corn. Proline betaine (=stachydrine) and glycine betaine were also detected in these extracts as major components. This constitutes the first report of chaetoglobosin M, chaetoglobosin O, proline betaine, or glycine betaine from S. maydis, and the first record of diplodiatoxin, diplonine, proline betaine, glycine betaine, or chaetoglobosins M, O, or K being associated with a natural field outreak of S. maydis ear rot.     

Anti-influenza (H1N1) potential of leaf and stem bark extracts of selected medicinal plants of South India

Variations in antioxidant and anti-viral activities (against Influenza AP/R/8 (H1N1) virus) between the leaves and stem bark of selected medicinal plants were studied. Malin Darby canine kidney (MDCK) cells were used for the viral infection and the antiviral activity of the extracts was studied using sulphorhodamine B (SRB) assay. The stem bark of the plants including Strychnos minor, Diotacanthus albiflorus, Strychnos nux-vomica and Chloroxylon swietenia showed higher flavonoid contents as well as 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activity when compared with their leaves. In case of 1,1-diphenyl-2-picrylhydrazyl (DPPH) activity, the stem bark of S. nux-vomica and leaf extract of C. swietenia showed the highest activity. Based on the IC₅₀ values, the stem bark extracts of Cayratia pedata (20.5 μg/mL) and S. minor (22.4 μg/mL) showed high antiviral activity. In the mean-time S. nux-vomica, C. swietenia and C. swietenia bark extracts showed cytotoxicity to the MDCK cells. When comparing the stem bark and leaves the content of gallic acid, ferulic acid, o-coumaric acid, total flavonoids (TFC) and total phenols (TPC) was higher in stem bark and hence their anti-viral activity was high. Further study based on the metabolites against H1N1 can reveal the potential of therapeutic compounds against the viral disease.     

Triterpenoid,coumarin and quinone constituents of eleven Diospyros species (Ebenaceae)

From the bark and/or timber extracts of Diospyros hirsuta, D. moonii, D. quaesita, D. spinescens, D. thwaitesii and D. walkeri, the following compounds have been isolated; lupeol, betulin, betulinic acid sitosterol, taraxerol, taraxerone, ursolic acid, oleanolic acid scopoletin, plumbagin, elliptinone, diospyrin and diosindigo A. TLC examination of the bark and timber extract of D. acuta, D. chaetocarpa, D. oblongifolia, D. oppositifolia and D. rheophytica is reported. Lupeol betulin, oleanolic acid and sitosterol have been isolated from the fruit of D. oblongfolia.     

The antiradical activity of plant extracts and their health-improving and prophylactic combinations with a phosholipid complex

Using the chemiluminescence method, the effective concentration of antioxidants (AO) and their antiradical activity (ARA) have been measured for 13 plant extracts. All extracts demonstrated higher ARA than that of the synthetic antioxidant ionol. The highest ARA was found in extracts from Larix dahurica, Hypericum perforatum, Potentilla fruticosa, Aronia melanocarpa, and Rhaponticum carthamoides. Synergistic action was found for combinations of extracts from Aronia + Rhaponticum, Larix + Hibiscus, and Schizandra + Aronia; the synergistic effect β was 38, 33, and 22%, respectively. This effect may be attributed to compounds present in these extracts. Phospholipids (the phospholipid complex Lipoid S40) lacking any antioxidant effect alone, showed a potent synergistic effect in combination with the Aronia extract (β = 60%) and the Silybum extract (β = 41%). Combinations of plant extracts with the phospholipids complex potentiated their inhibitory activity by increasing the induction period. Clinical trials have demonstrated, the combinations used may be recommended as an additional component in the complex therapeutic treatment of such chronic diseases as cardiovascular and hepatobiliary and also as an individual prophylactic agent.     

Neurohistochemistry in molluscan species: focus on extracellular matrix

Carbohydrate polymers with acid?Cbase properties of extracellular matrix (ECM) have a critical impact on guidance, migration and synaptic transmission. The present study showed acid?Cbase properties of ECM stained with histochemical techniques. The central nervous system (CNS) of mature terrestrial slug Semperula maculata and freshwater bivalve Lamellidens corrianus was selected to visualize sulphated and carboxylic strong acid groups. Protein and carbohydrate molecules were found scattered in three different regions, viz. periganglionic sheath, along with the basement lamina, interperikaryal space including the perineuronal net, and neuropil, in both the molluscan species. Sulphated proteoglycans were abundant in periganglionic sheath of S. maculata and also occurred in the neuropil of L. corrianus. The interperikaryal space contained carboxyl groups along with hyaluronic acid, and neuropil contained abundant glycogen moiety in both species. Comparing the SDS?CPAGE of the neural tissue extracts revealed protein molecules that were found in the same fraction. The ECM of neuronal cells from S. maculata and L. corrianus presented different kinds of glycoproteins and proteoglycans, which showed its morphological as well as habitual differentiation.