Novel Cell Permeable Peptide Based on Multi Basic Furin-dependent Cleavage Site of Respiratory Syncytial Virus Fusion Protein

Cell permeable peptide (CPP) is able to transport itself or conjugated molecules such as nucleotides, peptides, and proteins into cells. Since short peptide of human immunodeficiency virus-1 Tat has been discovered as CPP, it has been continuously studied for their ability to transport heterologous cargoes into cells. In this study, we have focused on the fusion protein of respiratory syncytial virus (RSV), which has six basic amino acids in multi basic furin-dependent cleavage site (MBFCS) required to be cationic CPP. To develop more efficient CPP, the sequence, which linked two MBFCS, was synthesized (called RS-CPP). To assess cell permeable efficiency of RS-CPP or MBFCS, the peptides was conjugated with fluorescein isothiocyanate, and cell permeable efficiency was measured by fluorescence-activated cell sorting. Cell permeability of RS-CPP or MBFCS was increased in a dose-dependent manner, but RS-CPP showed more efficient cell permeability than MBFCS in MDCK, HeLa, Vero E6, and A549 cells. To evaluate whether RS-CPP can transport its conjugated functional peptide (VIVIT) in CD8+ T cell, it was confirmed that IL-2 and β-galactosidase expression were significantly inhibited through selective block of nuclear factor activated T-cell. To investigate endocytic pathways, Cre-mediated DNA recombination (loxP-STOP-loxP-LacZ reporter system) was investigated with divergent endocytosis inhibitors in TE671 cells, and RS-CPP endocytosis is occurred via binding cell surface glycosaminoglycan and clathrin-mediated endocytosis, or macropinocytosis. These results indicated that RS-CPP could be a novel cationic CPP, and it would help understanding for delivery of biologically functional molecules based on viral basic amino acids.

Characterisation of genotype VII Newcastle disease virus (NDV) isolated from NDV vaccinated chickens, and the efficacy of LaSota and recombinant genotype VII vaccines against challenge with velogenic NDV

A Newcastle disease virus (NDV) isolate designated IBS002 was isolated from a commercial broiler farm in Malaysia. The virus was characterised as a virulent strain based on the multiple basic amino acid motif of the fusion (F) cleavage site 112RRRKGF117 and length of the C-terminus extension of the hemagglutinin-neuraminidase (HN) gene. Furthermore, IBS002 was classified as a velogenic NDV with mean death time (MDT) of 51.2 h and intracerebral pathogenicity index (ICPI) of 1.76. A genetic distance analysis based on the full-length F and HN genes showed that both velogenic viruses used in this study, genotype VII NDV isolate IBS002 and genotype VIII NDV isolate AF2240-I, had high genetic variations with genotype II LaSota vaccine. In this study, the protection efficacy of the recombinant genotype VII NDV inactivated vaccine was also evaluated when added to an existing commercial vaccination program against challenge with velogenic NDV IBS002 and NDV AF2240-I in commercial broilers. The results indicated that both LaSota and recombinant genotype VII vaccines offered full protection against challenge with AF2240-I. However, the LaSota vaccine only conferred partial protection against IBS002. In addition, significantly reduced viral shedding was observed in the recombinant genotype VII-vaccinated chickens compared to LaSota-vaccinated chickens.

Study on basic amino acid contents in Dendrobium officinale / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the contents of 16 basic amino acid and find out the variation of them in Dendrobium officinale with different germplasms and physiological ages, and then provide scientific basis for the quality evaluation and the breeding of D. officinale.</p><p><b>METHOD</b>Thirty-three samples with 1-3 ages were collected from cultivated fields of Zhejiang. The samples were acid hydrolyzed, and then 16 basic amino acid contents of samples were determined by amino acid analyzer.</p><p><b>RESULT</b>The average contents of 7 necessary amino acid were in 0.28 - 2.96 mg x g(-1), the average contents of other 9 basic amino acid were in 0.53 - 4.20 mg x g(-1). The contents of many amino acids were impacted by germplasms significantly, and contents of several amino acids were impacted by physiological ages significantly.</p><p><b>CONCLUSION</b>There were rich basic amino acids in D. officinale. The breeding of D. officinale can increase the contents of essential amino acids and other basic amino acids. The relations among physiological age and amino acid contents were as follows: three years > two years > one year. The contents of Asp and Tyr have significantly negative correlation with magnesium, the content of Pro has significantly positive correlation with copper.</p>

Effect of glycemic control and hypertension on Serum basic amino acids in type II diabetes mellitus

Abnormal insulin secretion has been considered the main cause of changes in carbohydrate lipid and protein metabolism in diabetes mellitus [DM] with increased muscle proteolysis and amino acid mobilization to study the effect of glycemic control and hypertension, on three basic amino acids arginine [Arg.] lysine [Lys] and histidine [His] in type II DM. Fifty five patients with type II diabetes mellitus [DM] were included in the study. They were 39 nortnotensive and 16 hypertensives with different grades of glycemic control as revealed by the level of glycated Hb [HbAk]. Thirty eight were with poor control; HbAlc > 8.5%, [27 normotensives and II hypertensives] while 17 were with acceptable range of control, HbAlc 6.8 - 8.5% [12 normotensives and 5 hypertensives]. Forty non diabetic subjects [28 normotensives and 12 hypertensives] were also included in the study as a control group. Glycatcd Hb [HbAk] was measured by affinity resin column chromatography and serum amino acid analysis was done by reversed high performance liquid chromatography. The results show that type II DM had caused a significant reduction in serum Arg [a precursor of nitric oxide which has an important role in the reduction of diabetic complications] with a significant elevation in serum Lys.while no change in serum His could be seen. Hypertension, on the other hand, has caused a significant elevation in serum His in both diabetics and non diabetics. Serum Arg was not changed while a significant elevation in serum Lys. was found in the non diabetic hypertensive subjects which disappeared in the diabetic hypertensive patients. No effect of the glycemic control on the concentration of any of the three amino acids could be detected in the diabetics of the present study. The possible suggestions and explanations for these events are mentioned

Efficacy of Panipenem versus Imipenem against Clinical Isolate of Pseudomonas aeruginosa in a Two-Compartment Artificial Capillary Model

BACKGROUND: Panipenem (PAPM) is a new carbapenem which has an enhanced broad spectrum activity against both gram-positive and negative organisms. However, its activities in vitro against Pseudomonas aeruginosa are still under controversy. The aim of this study was to compare the activity of PAPM with those of imipenem (IMPM) against clinical isolates of P. aeruginosa using in vitro kinetic model and to evaluate the differences according to the quantity of basic amino acid in media. MATERIALS AND METHODS: Using a clinical isolate of P. aeruginosa (SGP14) from blood, an in vitro 2-compartment artificial capillary model based on a dialyzer unit was prepared. Antibiotics were given as a bolus q12 hrs for 48 hrs. Simulated doses and frequencies of PAPM and IMPM were 500 mg q12 hrs as approved by Korea Food and Drug Administration. Muller-Hinton broth (MHB) and minimal broth Davis (MBD) were used as culture media and we divided the experiments into 4 groups [PAPM (MHB), PAPM (MBD), IMPM (MHB), IMPM (MBD)]. At 0, 1, 2, 4, 6, 8, 12, 24, 32, and 48 h, samples were removed from peripheral compartment and viable bacterial counts were measured. RESULTS: The susceptibility of PAPM and IMPM for SGP14 were 64 and 2 ug/mL in MHB and 4 and 2 ug/mL in MBD, respectively. Up until 12 hours, changes in bacterial colony counts were not significantly different (P=0.073) for each group. However among the four groups, PAPM (MHB) showed the least changes compared with PMPM (MBD), IMPM (MBD). The largest decrement of colony during 48 hours was observed with PMPM (MBD), followed by IMPM (MHB) or IMPM (MBD), and PAPM (MHB) in decreasing order (P=0.00). There were no differences between IMPM (MHB) and IMPM (MBD) as for the change in colony counts. CONCLUSIONS: The bactericidal activities of panipenem against the clinical isolate of P. aeruginosa was similar (at 12 h) or superior (at 48 h) to that of imipenem in an in vitro 2-compartment artificial capillary model using minimal broth to simulate human serum drug concentrations.

Efficacy of Panipenem versus Imipenem against Clinical Isolate of Pseudomonas aeruginosa in a Two-Compartment Artificial Capillary Model

BACKGROUND: Panipenem (PAPM) is a new carbapenem which has an enhanced broad spectrum activity against both gram-positive and negative organisms. However, its activities in vitro against Pseudomonas aeruginosa are still under controversy. The aim of this study was to compare the activity of PAPM with those of imipenem (IMPM) against clinical isolates of P. aeruginosa using in vitro kinetic model and to evaluate the differences according to the quantity of basic amino acid in media. MATERIALS AND METHODS: Using a clinical isolate of P. aeruginosa (SGP14) from blood, an in vitro 2-compartment artificial capillary model based on a dialyzer unit was prepared. Antibiotics were given as a bolus q12 hrs for 48 hrs. Simulated doses and frequencies of PAPM and IMPM were 500 mg q12 hrs as approved by Korea Food and Drug Administration. Muller-Hinton broth (MHB) and minimal broth Davis (MBD) were used as culture media and we divided the experiments into 4 groups [PAPM (MHB), PAPM (MBD), IMPM (MHB), IMPM (MBD)]. At 0, 1, 2, 4, 6, 8, 12, 24, 32, and 48 h, samples were removed from peripheral compartment and viable bacterial counts were measured. RESULTS: The susceptibility of PAPM and IMPM for SGP14 were 64 and 2 ug/mL in MHB and 4 and 2 ug/mL in MBD, respectively. Up until 12 hours, changes in bacterial colony counts were not significantly different (P=0.073) for each group. However among the four groups, PAPM (MHB) showed the least changes compared with PMPM (MBD), IMPM (MBD). The largest decrement of colony during 48 hours was observed with PMPM (MBD), followed by IMPM (MHB) or IMPM (MBD), and PAPM (MHB) in decreasing order (P=0.00). There were no differences between IMPM (MHB) and IMPM (MBD) as for the change in colony counts. CONCLUSIONS: The bactericidal activities of panipenem against the clinical isolate of P. aeruginosa was similar (at 12 h) or superior (at 48 h) to that of imipenem in an in vitro 2-compartment artificial capillary model using minimal broth to simulate human serum drug concentrations.