Lenediamine complexes with gold was substantially milder than cisplatin with no evidence of apoptosis or necrosis in the entire series of animals receiving a drug dose of 32.2 mg/kg for 14 days.ConclusionsGold (III) compound [Au(en)Cl2]Cl in sub-acute toxicity study, produced less renal and hepatic 1326631 toxicity as compared to other clinically established antineoplastic drugs. In the entire series of animals, no renal tubular necrosis was seen. Mild pyelitis and congestion dominated the histopathological picture. In hepaticRenal and Hepatic Toxicity of a Gold (III) Compoundtissue, ballooning degeneration of varied extent and severity prevailed in the drug dosed animals with no evidence of hepatocytic degeneration and necrosis.Author ContributionsConceived and designed the experiments: AA AI AMMA. Performed the experiments: AA DT MS. Analyzed the data: AA MS. Contributed reagents/materials/analysis tools: AA AI AMMA DT MS. Wrote the paper: AA AI MS. Designing and writing grant proposal: AI AA AMMA MS DT. 4 IBP cost Developing the drug: AI AMMA. Treating the animals: AMMA. Preparing the tissue, histological evaluation: AA MS DT. Analysis and preparing manuscript: AA AI AMMA MS DT.AcknowledgmentsWe acknowledge services of Mrs Khalda Al Johy, Mrs Zainab Al Najar, Mr Shakir Ahmad and Mrs Maria Rosario Lazaro in conducting the laboratory work.
Quantitative PCR (qPCR) is a sensitive and reliable 1379592 method used to quantify the number of target gene copies in a given sample. The accuracy of absolute quantification relies on the use of standards of known copy numbers run in the same experiment as the sample(s) being analyzed [1]. In environmental and industrial microbiology, microbial counts can be rapidly deduced using molecular methods based on known numbers of 16S rRNA genes or specific functional genes present in the genome [2]. The 16S rRNA gene is the most widely used tool for assessing microbial diversity and numbers in environmental samples using PCR-based methods [1,3]. The ,1500 base pair (bp) full-length 16S rRNA gene sequence contains conserved regions that are flanked by hyper-variable regions, and it is the degree of variation within these hyper-variable regions that distinguishes between microbial taxa at various classification levels [4,5]. Therefore, taxa-specific 16S rRNA gene primer sets coupled with SYBR chemistry is the most cost-effective method to target and quantify the maximum number of microbes at a given classification level within a test sample Cucurbitacin I reviewed in [6]. A recent report has shown that supercoiled plasmid DNA used to generate standard curves grossly overestimated the number of pcna gene copies by 8-fold using the microalgae eukaryotic system[7]. Lin et al. [8] reported a similar finding, with a 3-fold increase in the NK603/zSSIIb gene(s) using the eukaryotic system, maize. Interestingly, little attention has been paid to the effect of circularTable 1. List of primers used to amplify the 16S rRNA gene.Primer Cloning fD1 rP2 Arc8f Arc1492r qPCR 27f 338r Arc8f Arc344rTaxa/ Direction Domain Sequence (59-39)ReferenceForward Reverse Forward ReverseBacteriaAGAGTTTGATCCTGGCTCAG ACGGCTACCTTGTTACGACTT[5]Archaea TCCGGTTGATCCTGCC GGCTACCTTGTTACGACTT[13]Forward Reverse Forward ReverseBacteriaAGAGTTTGATCMTGGCTCAG GCTGCCTCCCGTAGGAGT[14]Archaea TCCGGTTGATCCTGCC TCGCGCCTGCTGCICCCCGT[15]doi:10.1371/journal.pone.0051931.tEffect of qPCR Standards on 16S Gene Estimatesplasmid standards in bacterial and archaeal systems which commonly have genomes and pl.Lenediamine complexes with gold was substantially milder than cisplatin with no evidence of apoptosis or necrosis in the entire series of animals receiving a drug dose of 32.2 mg/kg for 14 days.ConclusionsGold (III) compound [Au(en)Cl2]Cl in sub-acute toxicity study, produced less renal and hepatic 1326631 toxicity as compared to other clinically established antineoplastic drugs. In the entire series of animals, no renal tubular necrosis was seen. Mild pyelitis and congestion dominated the histopathological picture. In hepaticRenal and Hepatic Toxicity of a Gold (III) Compoundtissue, ballooning degeneration of varied extent and severity prevailed in the drug dosed animals with no evidence of hepatocytic degeneration and necrosis.Author ContributionsConceived and designed the experiments: AA AI AMMA. Performed the experiments: AA DT MS. Analyzed the data: AA MS. Contributed reagents/materials/analysis tools: AA AI AMMA DT MS. Wrote the paper: AA AI MS. Designing and writing grant proposal: AI AA AMMA MS DT. Developing the drug: AI AMMA. Treating the animals: AMMA. Preparing the tissue, histological evaluation: AA MS DT. Analysis and preparing manuscript: AA AI AMMA MS DT.AcknowledgmentsWe acknowledge services of Mrs Khalda Al Johy, Mrs Zainab Al Najar, Mr Shakir Ahmad and Mrs Maria Rosario Lazaro in conducting the laboratory work.
Quantitative PCR (qPCR) is a sensitive and reliable 1379592 method used to quantify the number of target gene copies in a given sample. The accuracy of absolute quantification relies on the use of standards of known copy numbers run in the same experiment as the sample(s) being analyzed [1]. In environmental and industrial microbiology, microbial counts can be rapidly deduced using molecular methods based on known numbers of 16S rRNA genes or specific functional genes present in the genome [2]. The 16S rRNA gene is the most widely used tool for assessing microbial diversity and numbers in environmental samples using PCR-based methods [1,3]. The ,1500 base pair (bp) full-length 16S rRNA gene sequence contains conserved regions that are flanked by hyper-variable regions, and it is the degree of variation within these hyper-variable regions that distinguishes between microbial taxa at various classification levels [4,5]. Therefore, taxa-specific 16S rRNA gene primer sets coupled with SYBR chemistry is the most cost-effective method to target and quantify the maximum number of microbes at a given classification level within a test sample reviewed in [6]. A recent report has shown that supercoiled plasmid DNA used to generate standard curves grossly overestimated the number of pcna gene copies by 8-fold using the microalgae eukaryotic system[7]. Lin et al. [8] reported a similar finding, with a 3-fold increase in the NK603/zSSIIb gene(s) using the eukaryotic system, maize. Interestingly, little attention has been paid to the effect of circularTable 1. List of primers used to amplify the 16S rRNA gene.Primer Cloning fD1 rP2 Arc8f Arc1492r qPCR 27f 338r Arc8f Arc344rTaxa/ Direction Domain Sequence (59-39)ReferenceForward Reverse Forward ReverseBacteriaAGAGTTTGATCCTGGCTCAG ACGGCTACCTTGTTACGACTT[5]Archaea TCCGGTTGATCCTGCC GGCTACCTTGTTACGACTT[13]Forward Reverse Forward ReverseBacteriaAGAGTTTGATCMTGGCTCAG GCTGCCTCCCGTAGGAGT[14]Archaea TCCGGTTGATCCTGCC TCGCGCCTGCTGCICCCCGT[15]doi:10.1371/journal.pone.0051931.tEffect of qPCR Standards on 16S Gene Estimatesplasmid standards in bacterial and archaeal systems which commonly have genomes and pl.