The effect of condensed tannins fromLotus pedunculatusandLotus corniculatuson the growth of proteolytic rumen bacteria in vitro and their possible mode of action

2001 ◽  
Vol 47 (7) ◽  
pp. 626-633 ◽  
Author(s):  
A L Molan ◽  
G T Attwood ◽  
B R Min ◽  
W C McNabb
Keyword(s):  
Mode Of Action ◽  
Condensed Tannins ◽  
Large Subunit ◽  
Rumen Bacteria ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
Bisphosphate Carboxylase ◽  
Butyrivibrio Fibrisolvens ◽  
Significant Difference

Five strains of proteolytic rumen bacteria were treated with condensed tannins (CT) purified from Lotus pedunculatus and Lotus corniculatus to investigate their effect on the growth of these bacteria in vitro. Streptococcus bovis NCFB 2476, Eubacterium sp. C124b, Prevotella bryantii B14, Butyrivibrio fibrisolvens H17c, and Clostridium proteoclasticum B316Twere tested against 200, 400, and 600 µg CT·mL–1extracted from L. pedunculatus and L. corniculatus. In the absence of CT, all bacterial strains showed typical growth and reached maximum optical density (OD) after 6–8 h of incubation in a plant protein medium. Growth of Eubacterium sp., P. bryantii, and B. fibrisolvens was inhibited (P < 0.01–0.001) more by the CT from L. pedunculatus than by the CT from L. corniculatus. All strains continued to grow in the presence of 200 µg·mL–1of the CT from L. pedunculatus, but attained significantly (P < 0.05–0.01) lower maximum OD600values than (minus CT) controls, except for S. bovis. At 400 and 600 µg·mL–1, the addition of CT from L. pedunculatus inhibited (P < 0.05–0.001) the growth of all bacterial strains tested compared with controls. The growth of Eubacterium sp. and P. bryantii was stimulated for the first 4–6 h of incubation (P < 0.001) by 200 µg·mL–1of CT from L. corniculatus, but then declined leading to a significant difference in OD values compared with the controls. At 400 µg·mL–1, the CT from L. corniculatus reduced (P < 0.05–0.01) the growth of all strains except S. bovis, while 600 µg·mL–1inhibited (P < 0.01–0.001) the growth of all strains. To study the mechanism of CT action, the degradation of the large subunit (LSU) of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; Fraction 1 Leaf protein) was followed after bacterial cells or Rubisco were preincubated with CT extracted from L. corniculatus and L. pedunculatus. Both preincubations decreased LSU degradation, but they differed in their response to polyethylene glycol (PEG) addition. Addition of PEG to CT–Rubisco preincubations negated the effects of CT, while PEG addition to CT–bacteria preincubations did not. This implies that the CT–bacterial interaction is stronger than the CT–Rubisco interaction or the interaction is of a different type. Also, L. pedunculatus CT reduced the degradation of the LSU to a greater extent than the CT from L. corniculatus when preincubated with bacteria.Key words: condensed tannins, growth, in vitro, proteolytic rumen bacteria, mode of action, Rubisco.

scholarly journals Characterization of the gene and mRNA of the large subunit of ribulose-1,5-bisphosphate carboxylase in pea plants.

1983 ◽  
Vol 3 (4) ◽  
pp. 587-595 ◽  
Author(s):  
K K Oishi ◽  
K K Tewari
Keyword(s):  
Immunoglobulin G ◽  
Large Subunit ◽  
Rrna Genes ◽  
Mapping Technique ◽  
Affinity Column ◽  
Bisphosphate Carboxylase ◽  
Bamhi Fragment ◽  
Dna Fragment

mRNA coding for the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase was obtained by fractionating chloroplast polysomes on an affinity column, using anti-ribulose-1,5-bisphosphate carboxylase immunoglobulin G. Approximately 20% of the polysomal RNA specifically bound to the affinity column. LS mRNA was also isolated by fractionating chloroplast polysomal RNA on sucrose gradients. The LS mRNA fraction was identified by translation in vitro followed by immunoprecipitation with anti-ribulose-1,5-bisphosphate carboxylase immunoglobulin G. Labeled LS mRNA was hybridized to a genomic digests of pea chloroplast DNA. The LS gene was localized on a 3.55-kilobase pair BamHI fragment in SalI-SmaI DNA fragment 4. The BamHI fragment containing the LS gene was cloned, and a restriction endonuclease map was constructed. The LS gene was localized on a 1.9-kbp KpnI-EcoRI fragment. The LS gene was analyzed by electron microscopy, using the R loop mapping technique. LS mRNA was colinear with the gene, and its size was 1.35 +/- 0.2 kilobase pairs. When the LS mRNA was analyzed on methylmercury agarose gels, it comigrated with the 16S rRNA. The direction of transcription of the LS gene was in the same direction as that of the rRNA genes.

scholarly journals IN VITRO ANTIOXIDANT AND IN VIVO ANTIDIABETIC ACTIVITY OF TWO POTENTIAL PROBIOTIC ENTEROCOCCUS SPP. ON ALLOXAN-INDUCED DIABETIC RATS

2021 ◽  
pp. 94-98
Author(s):  
KAMNI RAJPUT ◽  
RAMESH CHANDRA DUBEY
Keyword(s):  
Diabetic Rats ◽  
Antidiabetic Activity ◽  
Control Group ◽  
Albino Rats ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
Animal Group ◽  
Enterococcus Spp

Objective: In vitro antioxidant activity, in vivo antidiabetic property and intestinal attachment by two potential probiotic bacterial strains, namely, Enterococcus faecium and Enterococcus hirae were studied using albino rats. Methods: Antioxidant the activity was assessed using 2,2-Diphenyl-1-picrylhydrazyl radicals scavenging assay. Alloxan was administered intraperitoneally to induce diabetic conditions in experimental rats. Animals were treated with oral administration of Enterococcus spp., such as E. faecium, and E. hirae isolated from goat and sheep milk. The control animal group received normal saline for the same days. Glibenclamide drug was used as a positive control against probiotic bacterial cells. Results: However, administration of probiotic bacterial strains E. faecium and E. hirae, in albino rats significantly (p<0.05) at varying doses lowered blood glucose levels in diabetic rats as compared to the diabetic control group. Both the species of Enterococcus increased the bodyweight of experimental rats. However, E. faecium was the best antidiabetic strain having the antioxidant activities also in comparison to E. hirae. The attachment of probiotic bacterial cells E. faecium on the rat’s intestine wall against pathogens was examined. Furthermore, E. faecium showed its aggregation with pathogens by attachment of the intestines of albino rats. This showed that both the bacterial strains exhibited in vivo antidiabetic effect. Conclusion: The results of this study showed that probiotic bacteria possess antioxidant, antidiabetic activities, and attachment of intestine.

scholarly journals In vitro synthesis and processing of a maize chloroplast transcript encoded by the ribulose 1,5-bisphosphate carboxylase large subunit gene.

1985 ◽  
Vol 5 (10) ◽  
pp. 2733-2745 ◽  
Author(s):  
L Hanley-Bowdoin ◽  
E M Orozco ◽  
N H Chua
Keyword(s):  
Transcription Initiation ◽  
Large Subunit ◽  
Plastid Dna ◽  
Subunit Gene ◽  
Coding Region ◽  
Protein Coding ◽  
Bisphosphate Carboxylase ◽  
Transcription In Vitro ◽  
Maize Chloroplast

The large subunit gene (rbcL) of ribulose 1,5-bisphosphate carboxylase was transcribed in vitro by using maize and pea chloroplast extracts and a cloned plastid DNA template containing 172 base pairs (bp) of the maize rbcL protein-coding region and 791 bp of upstream sequences. Three major in vitro RNA species were synthesized which correspond to in vivo maize rbcL RNAs with 5' termini positioned 300, 100 to 105, and 63 nucleotides upstream of the protein-coding region. A deletion of 109 bp, including the "-300" 5' end (the 5' end at position -300), depressed all rbcL transcription in vitro. A plasmid DNA containing this 109-bp fragment was sufficient to direct correct transcription initiation in vitro. A cloned template, containing 191 bp of plastid DNA which includes the -105 and -63 rbcL termini, did not support transcription in vitro. Exogenously added -300 RNA could be converted to the -63 transcript by maize chloroplast extract. These results established that the -300 RNA is the primary maize rbcL transcript, the -63 RNA is a processed form of the -300 transcript, and synthesis of the -105 RNA is dependent on the -300 region. The promoter for the maize rbcL gene is located within the 109 bp flanking the -300 site. Mutagenesis of the 109-bp chloroplast sequence 11 bp upstream of the -300 transcription initiation site reduced rbcL promoter activity in vitro.

Effects of heavy metals and other trace elements on the fermentative activity of the rumen microflora and growth of functionally important rumen bacteria

1978 ◽  
Vol 24 (3) ◽  
pp. 298-306 ◽  
Author(s):  
C. W. Forsberg
Keyword(s):  
Trace Elements ◽  
Rumen Fluid ◽  
Inhibitory Effect ◽  
Rumen Bacteria ◽  
Inhibitory Effects ◽  
Butyrivibrio Fibrisolvens ◽  
Fermentative Activity ◽  
Rumen Microflora ◽  
High Concentrations

The inhibitory effects of high concentrations of essential and non-essential trace elements were tested on the rumen microflora using the rate of fermentation in vitro as the assay. The elements (and the concentration causing 50% inhibition) in decreasing order of toxicity were Hg2+ (20 μg/ml), Cu2+ (21 μg/ml), Cr6+ (70 μg/ml), Se4+ (73 μg/ml), Ni2+ (160 μg/ml), Cd2+ (175 μg/ml), As3+ (304 μg/ml), and As5+ (1610 μg/ml). The elements tested that were either weak or non-inhibitory at concentrations greater than 400 μg/ml included Zn2+, Cr2+, Fe2+, Mn2+, Pb2+, and Co2+. Methylmercury was as inhibitory as mercuric chloride to the fermentation. When the inhibitory effect of Cd2+ was tested on separated bacterial and protozoal fractions, it was more inhibitory to the bacteria. The inhibitory effects of trace elements were also determined for a number of axenic cultures of rumen bacteria. The bacteria which most frequently exhibited the greatest sensitivity were Bacteroides succinogenes, Ruminococcus albus, Bacteroides amytophilus, and Eubacterium ruminantium. Those often exhibiting intermediate sensitivities included Butyrivibrio fibrisolvens, Selenomonas niminantium, and Megasphera elsdenii, while Streptococcus bovis was very refractory to all elements tested. Rumen fluid provided a modest protective effect for the bacteria.

scholarly journals In vivo capture of bacterial cells by remote guiding

2021 ◽  
Author(s):  
Iaroslav A. Rybkin ◽  
Sergey I. Pinyaev ◽  
Olga A. Sindeeva ◽  
Sergey V. German ◽  
Maja Koblar ◽  
...  
Keyword(s):  
Genetic Material ◽  
Local Concentration ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
Toxic Side Effect ◽  
High Concentrations ◽  
Strong Barrier ◽  
Growth Distribution

Recently, it has been shown that several bacterial strains can be very efficient in cancer treatment since they possess many important properties such as self-targeting, ease of detection, sensing and toxicity against tumors. However, there are only a few relevant candidates for such an approach, as targeting and detection one of the biggest challenges as well as there are many limitations in the use of genetic approaches. Here, it is proposed the solution that enables surface modification of alive bacterial cells without interfering with their genetic material and potentially reduces their toxic side effect. By the electrostatic interaction fluorescently labeled polyelectrolytes (PEs) and magnetite nanoparticles (NPs) were deposited on the bacterial cell surface to control the cell growth, distribution and detection of bacteria. According to the results obtained in vivo, by the magnet entrapment of the modified bacteria the local concentration of the cells was increased more than 5 times, keeping the high concentrations even when the magnet is removed. Since the PEs create a strong barrier, in vitro it was shown that the division time of the cells can be regulated for better immune presentation.

scholarly journals In vitro Evaluationof the Antimi- crobial Activity of a Topical Skin Preparation Containing 0.1% Polyhexanide vs a Topical Skin Prepa-ration Containing 1% Silver Sulfadiazine

2021 ◽  
Vol 6 (2) ◽  
pp. 1-7
Author(s):  
Barbara Maglione ◽  
Keyword(s):  
Antibacterial Activity ◽  
Positive Control ◽  
Negative Control ◽  
Silver Sulfadiazine ◽  
Bacterial Strains ◽  
Skin Preparation ◽  
E Coli ◽  
Topical Cream ◽  
Significant Difference

Aim: The effective in vitro antibacterial activity on Staphylococcus aureus (S.aureus), Pseudomonas aeruginosa (P.aeruginosa), Klebsiella pneumoniae (K.pneumoniae),Escherichia coli (E.Coli) and the combination of S.aureus and K. pneumonia of a topical cream based on 0.1% polyhexanidewas compared to a topical cream based on 1% silver sulfadiazine.A topical cream containing 0,1% gentamicin was used as a positive control and a white blank topical cream was used as negative control. Materials and Methods: The in vitro antibacterial activities were determined by agar well-diffusion assay. Two-way Analysis of Variance (ANOVA) was used to test, by calculation of P-values, for significant antiseptic activity in bacteria treated with 0.1% polyhexanide topical cream compared to 1% silver sulfadiazine and to the negative and positive controls. Results: Among the derivatives tested, all the active topical creams analyzed were able to reduce microbial strains. The topical cream based on 0.1% polyhexanide showed a significantly higher antibacterial efficacy in comparison to the topical cream based on 1% silver sulfadiazine on S. aureus and K. pneumonia and on the combination of S. aureus and K. pneumoniae,while no significant difference was detected between the antibacterial activity of the two topical creams against P. aeruginosa and E. coli. Conclusion: These results provide a further insight into the antibacterial activity of polyhexanide and its non-inferiority compared to silver sulfadiazine towards certain bacterial strains (P. aeruginosa and E. coli) and superiority towards other (S. aureus and K. pneumoniae)and support the use of 0.1% Polyhexanide topical preparation for the treatment of wounds that are infected or at risk of infection.

scholarly journals Inhibition of ribulose bisphosphate carboxylase assembly by antibody to a binding protein.

1986 ◽  
Vol 103 (4) ◽  
pp. 1327-1335 ◽  
Author(s):  
S Cannon ◽  
P Wang ◽  
H Roy
Keyword(s):  
Higher Plants ◽  
Binding Protein ◽  
Large Subunit ◽  
Ribulose Bisphosphate Carboxylase ◽  
Ribulose Bisphosphate ◽  
Dual Effect ◽  
Bisphosphate Carboxylase ◽  
Low Concentrations ◽  
The One

We have developed an assay to monitor in vitro the posttranslational assembly of the chloroplast protein, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO). Most of the newly synthesized 55-kD catalytic ("large") subunits of this enzyme occur in a 29S complex together with 60- and 61-kD "binding" proteins. When the 29S complex is incubated with ATP and MgCl2 it dissociates into subunits, and the formerly bound large subunits now sediment at 7S (still faster than expected for a monomer). Upon incubation at 24 degrees C, these large subunits assemble into RuBisCO. The minority of newly made large subunits which are not bound to the 29S complex also sediment at 7S. When endogenous ATP was removed by addition of hexokinase and glucose, the dissociation of the 29S complex was inhibited. Nevertheless, the 7S large subunits assembled into RuBisCO, and did so to a greater extent than in controls retaining endogenous ATP. Thus the 7S large subunits are also assembly competent, at least when ATP is removed. Apparently, in chloroplast extracts, ATP can have a dual effect on the assembly of RuBisCO: on the one hand, even at low concentrations it can inhibit incorporation of 7S large subunits RuBisCO; on the other hand, at higher concentrations it can lead to substantial buildup of the 7S large subunit pool by causing dissociation of the 29S complex, and stimulate overall assembly. At both high and zero concentrations of ATP, however, antibody to the binding protein inhibited the assembly of endogenous large subunits into RuBisCO. Thus it appears that all assembly-competent large subunits are associated with the binding protein, either in the 7S complex or in the 29S complex. The involvement of the binding protein in RuBisCO assembly may represent the first example of non-autonomous protein assembly in higher plants and may pose problems for the genetic engineering of RuBisCO from these organisms.

scholarly journals In-vitro Antimicrobial Activity of the Combined Effect of Kalanchoe Crenata and Vernonia Amygdalina on Salmonella Species

2020 ◽  
Author(s):  
Emmanuel Effah-Yeboah ◽  
Emmanuel Agyapong Asare ◽  
Gadafi Iddrisu Balali
Keyword(s):  
Growth Inhibition ◽  
Salmonella Typhi ◽  
Combined Effect ◽  
Diffusion Method ◽  
P Value ◽  
Bacterial Strains ◽  
Vernonia Amygdalina ◽  
Zone Of Inhibition ◽  
Significant Difference

Abstract IntroductionThe major breakthrough in the treatment of pathogenic diseases was the unearthing of naturally occurring antipathogenic agents or antibiotics. There have been upsurges in antibiotic-resistant strains of clinically important pathogens, which made way to the emergence of new-fangled bacterial strains that are multi-resistant. The major aim of scientists is to develop new antibiotics or other therapeutic strategies at a pace greater than that at which bacteria are developing resistance. Development of resistance to first-line antimicrobial therapies made way to recommendations for combination therapies for the treatment of some infections and some of this form of chemotherapy seems to be very successful.ObjectivesThis research was carried out the determine the effect of Kalanchoe crenata extract on salmonella Tyhi load. It was carried out to also assess the potency of the extract of Vernonia amygdalina on Salmonella typhi and also to ascertain the effect of the combined extract of Kalanchoe crenata and Vernonia amygdalina on salmonella typhi.MethodIn this research, Salmonella typhi was exposed to a crude extract of Kalanchoe crenata and Vernonia amygdalina and also the combination of the two extracts. Agar wells diffusion method was employed.ResultsThe combined effect was not sensitive to the Salmonella strain. The Salmonella strain was resistant to V. amygdalina than to K. crenata. K. cranata had the strongest activity against S. typhi with its highest zone of growth inhibition of 20 mm and lowest zone of inhibition of 7 mm while V. amygdalina produced consistent zone of growth inhibition of 5–6 mm; The combined effect produced a zone inhibition diameter only at the 100 mg/ml with zone of inhibition value of 14 mm. The subsequent lower concentrations did not show any activity against the microbes. At P-value = 0.05 two-way ANOVA statistics exhibited significant difference amongst the effects produced by the different extracts, though there were no substantial differences in the effects produced by the various concentrations.ConclusionThe salmonella strain was resistant to V. amygdalina than to K. crenata. At P-value = 0.05 there was a substantial difference in the sensitivity of the bacteria to the different extracts.

scholarly journals Investigating the Cytotoxic and Anti-Bacterial Activity of Commercially Available Local Anesthetics: An In-Vitro Analysis

2020 ◽  
Vol 29 (04) ◽  
pp. 185-192
Author(s):  
Eisha Imran ◽  
◽  
Faisal Moeen ◽  
Humayoon Satti ◽  
Lubna Rahman
Keyword(s):  
Local Anesthetic ◽  
Local Anesthetics ◽  
Bacterial Activity ◽  
Dilution Method ◽  
Human Mscs ◽  
Bacterial Strains ◽  
Significant Difference ◽  
Vitro Analysis ◽  
In Vitro Analysis

OBJECTIVE: To evaluate and compare the influence of three local anesthetic dental formulations manufactured in France (Septodont), Korea (Medicaine) and Pakistan (HD-Caine) in terms of cytotoxicity and anti-bacterial activities. METHODOLOGY: 90 commercially available local anesthetic cartridges of similar composition (2% lidocaine with epinephrine 100,000) viz. Septodont, Medicaine and HD-Caine were randomly collected from three different Pakistani cities and were assigned as Group S, Group M and Group H, respectively. The cartridges were further divided into three sub-groups each consisting of 10 cartridges to first evaluate cytotoxicity on Mesenchymal Stem Cells (MSCs) using a flow cytometer and secondly to investigate anti-bacterial activity by measuring zones of inhibition and through Broth Dilution Method against five bacterial strains. RESULTS: The results indicated that Septodont (94.5±0.1) and Medicaine (94.7±0.0) showed the highest viability percentage with no significant difference when the two were compared (P=0.6). HD-Caine (93.9±0.0) showed the least, being significantly (P<0.01) different from Septodont and Medicane. A statistically significant (P<0.05) difference was identified between the three study groups regarding the anti-bacterial activity. HD-Caine showed the highest anti-bacterial potential, followed by Medicaine and Septodont. CONCLUSION: Mild toxicity was observed by all the three groups in human MSCs, justifying their safe use in clinical practice. Additionally, Medicane and HD-Caine showed significant anti-bacterial activity indicating their possible use as sterile irrigants. KEYWORDS: Dental anesthesia, Lidocaine, Epinephrine, Antibacterial activity HOW TO CITE: Imran E, Moeen F, Satti H, Rahman L. Investigating the cytotoxic and anti-bacterial activity of commercially available local anesthetics: An in-vitro analysis. J Pak Dent Assoc 2020;29(4):185-192.

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