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OALib Journal期刊

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Basic peptide protamine exerts antimicrobial activity against periodontopathic bacteria——Growth inhibition of periodontopathic bacteria by protamine  [PDF]
Tadashi Miura, Keishi Iohara, Tetsuo Kato, Kazuyuki Ishihara, Masao Yoshinari
Journal of Biomedical Science and Engineering (JBiSE) , 2010, DOI: 10.4236/jbise.2010.311138
Abstract: Protamine was investigated for its antibacterial activity against the periodontal pathogens, Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans. We determined the minimum inhibitory concentrations of protamine and its hydrolysate and their bactericidal activity. Protamine inhibited the growth of all periodontopathic bacteria tested on agar plates. Protamine, which MIC was 6.3 × 10-7 g L-1, was most effective against P. gingivalis. The antibacterial effect of native protamine was higher than that of its hydrolysate. An ATP bioluminescence assay revealed that protamine showed bactericidal activity against P. gingivalis in a time-dependent manner. These results indicate that protamine could be candidate peptide for prevention of P. gingivalis infection.
Physiological Effects of Salmon Milt Nucleoprotein on Movement, Stress Tolerance and Lifespan of C. elegans  [PDF]
Hiroaki Shintani, Tsubasa Furuhashi, Hiroyuki Hano, Masaji Matsunaga, Koji Usumi, Norimasa Shudo, Kazuichi Sakamoto
Food and Nutrition Sciences (FNS) , 2012, DOI: 10.4236/fns.2012.31009
Abstract: In recent years, various physiological functions of salmon milt extract, which consists of nucleic acid and nucleoprotein, have been reported. The objective of this study is to analyze the physiological function and its mechanism of salmon milt extract (NG) on nematodes (C. elegans). The wild type nematode N2 strain was bred on the plate containing of NG for four days, and its body length increased depending on NG concentration. When nematodes were bred with NG for a longer period, average lifespan was increased, and survival rate was increased by up to 20%. Generally, the movement of nematodes decreases with longer breeding period (i.e. aging). Analysis of movement (both gross thrashing movement and local pumping movement) showed that NG suppressed this decrease f movement with aging. Furthermore, the deease of survival rate by heat stress and oxidative stress was suppressed by NG administration. Nile Red staining analysis showed that fat accumulation varied depending on the concentration of NG. RT-PCR analysis revealed that the mRNA expression levels of the stress resistance genes sod-3 and sod-4 were increased. These results indicated that NG administration increased the expression of stress-tolerance-related genes, promoted stress tolerance, increased movement and prolonged lifespan in nematode.
Flow Cytometry: A Novel Approach for Indirect Assessment of Protamine Deficiency by CMA3 Staining, Taking into Account the Presence of M540 or Apoptotic Bodies
Zohreh Fathi,Marziyeh Tavalaee,Mohammad Reza Deemeh,Mehrdad Modaresi
International Journal of Fertility & Sterility , 2011,
Abstract: Background: Chromomycin A3 (CMA3) staining, either by the slide method or fluorescence microscopy,is widely used for indirect assessment of protamine deficiency in a semen sample. Flow cytometry isthe most suitable tool to improve assessment accuracy, both in terms of statistical analysis and forprevention of observer variation. This study provides a simple procedure to account for merocyanine540 (M540) or apoptotic bodies, which result in underestimation of the percentage of CMA3 positivity,by using propidium iodide (PI) staining. Therefore, this study aims to evaluate the percentage of CMA3by PI staining to exclude M540 bodies that prevent underestimation of CMA3 staining.Materials and Methods: This study is an experimental study. Semen samples collected from 104infertile men who referred to the Andrology Unit of the Isfahan Fertility and Infertility Center wereinitially assessed according to World Health Organization (WHO) criteria. Samples were washedtwice with Ham’s. Each sample was divided into two portions, a control and the other processed fordensity gradient centrifugation (DGC). Each portion was assessed for CMA3 staining by both theslide and flow cytometry methods. Coefficients of correlation and student t-test were carried outusing the Statistical Package for the Social Studies (SPSS 11.5).Results: Detection of CMA3 staining was more appropriate with fluorescence detector 3 (FL-3)rather than fluorescence detector 2 (FL-2) in the evaluation of protamine deficiency to excludeM540 bodies.Conclusion: This study, for the first time, provides the basis for assessment of CMA3 staining forflow cytometry. However, since the maximum excitation for CMA3 is not covered by the 488 nmlaser, we recommend further experimentation using a flow cytometer with optimal excitation.
A New Histone Structure Which Binds DNA at Its Eight Subunit N-Termini  [PDF]
Ken Biegeleisen
Open Access Library Journal (OALib Journal) , 2016, DOI: 10.4236/oalib.1102386
Abstract: A new model for the nucleosome is presented. The histone octamer core is unchanged, but the location of the DNA is different. Since the highest number, and highest concentration of positively- charged amino acid residues is located not in the “superhelical ramp” of the octamer core, but rather in the domain of the eight histone subunit N-termini collectively, the DNA is therefore placed there. The role models for the protein and DNA structures in the N-terminal domain are taken from the comparable role models for protein and DNA in the protamine-DNA complex in sperm cells. The histone subunit N-termini are each modeled as beta-strands, with psi/phi values of approximately /﹣130.5° respectively, which gives a straight chain. The DNA is modeled according to the “straight ladder” model of Tai Te Wu. Each DNA phosphate group is bound to a lysine or arginine residue of histone by a 3 A salt bridge. The new model lends itself so readily to further models of higher-order chromatin structure that the problem shifts entirely, from one of deducing any higher-order structure at all, to one of distinguishing between several models which compete for our attention.
Improving the Stability of Insulin in Solutions Containing Intestinal Proteases in Vitro
Liefeng Zhang,Hui Jiang,Wenjie Zhu,Lin Wu,Lingling Song,Qiuyan Wu,Yong Ren
International Journal of Molecular Sciences , 2008, DOI: 10.3390/ijms9122376
Abstract: Degradation of insulin was studied in this work. Casein and protamine could obviously suppress degradation of insulin by intestinal enzymes, and could protect insulin from degradation by the mechanism of competition and combination with proteolysis enzyme. What is more, co-incubated with HP-β-CD-casein or HP-β-CD-protamine, most insulin was protected from degradation by intestinal enzymes. In addition, it was found that the complexation of insulin with HP-β-CD was characterized by UV absorption spectra. These results indicated that HP-β-CD, casein and protamine could offer some positive and useful results, and could protect insulin from degradation during their transit through the intestinal tract.
Assessment of a DNA vaccine encoding an anchored-glycosylphosphatidylinositol tegumental antigen complexed to protamine sulphate on immunoprotection against murine schistosomiasis
Nascimento, Eduardo JM;Amorim, Rosa V;Cavalcanti, André;Alves, Veruska F;Nakazawa, Mineo;Pereira, Valéria RA;Lucena-Silva, Norma;
Memórias do Instituto Oswaldo Cruz , 2007, DOI: 10.1590/S0074-02762007000100003
Abstract: protamine sulphate/dna complexes have been shown to protect dna from dnase digestion in a lipid system for gene transfer. a dna-based vaccine complexed to protamine sulphate was used to induce an immune response against schistosoma mansoni anchored-glycosylphosphatidylinositol tegumental antigen in balb/c mice. the protection elicited ranged from 33 to 44%. the spectrum of the elicited immune response induced by the vaccine formulation without protamine was characterized by a high level of igg (igg1> igg2a). protamine sulphate added to the dna vaccine formulation retained the green fluorescent protein encoding-plasmid longer in muscle and spleen. the experiments in vivo showed that under protamine sulphate effect, the scope of protection remained unchanged, but a modulation in antibody production (igg1= igg2a) was observed.
An efficient calcium phosphate nanoparticle-based nonviral vector for gene delivery
Liu YC, Wang T, He FL, Liu Q, Zhang DX, Xiang SL, Su SP, Zhang J
International Journal of Nanomedicine , 2011, DOI: http://dx.doi.org/10.2147/IJN.S17096
Abstract: n efficient calcium phosphate nanoparticle-based nonviral vector for gene delivery Original Research (6097) Total Article Views Authors: Liu YC, Wang T, He FL, Liu Q, Zhang DX, Xiang SL, Su SP, Zhang J Published Date April 2011 Volume 2011:6 Pages 721 - 727 DOI: http://dx.doi.org/10.2147/IJN.S17096 Yachun Liu1,2,*, Tao Wang1,*, Fangli He1,*, Qian Liu1,*, Dexi Zhang2, Shuanglin Xiang1, Shengpei Su2, Jian Zhang1 1Key Laboratory of Protein Chemistry and Developmental Biology, Ministry of Education of China, College of Life Sciences; 2Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research and Key Laboratory of Sustainable Resources Processing and Advanced Materials of Hunan Province, Ministry of Education of China, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, China *These authors contributed equally to this work Background: Smaller nanoparticles facilitate the delivery of DNA into cells through endocytosis and improve transfection efficiency. The aim of this study was to determine whether protamine sulfate-coated calcium phosphate (PS-CaP) could stabilize particle size and enhance transfection efficiency. Methods: pEGFP-C1 green fluorescence protein was employed as an indicator of transfection efficiency. Atomic force microscopy was used to evaluate the morphology and the size of the particles, and an MTT assay was introduced to detect cell viability and inhibition. The classical calcium phosphate method was used as the control. Results: Atomic force microscopy images showed that the PS-CaP were much smaller than classical calcium phosphate particles. In 293 FT, HEK 293, and NIH 3T3 cells, the transfection efficiency of PS-CaP was higher than for the classical calcium phosphate particles. The difference in efficiencies implies that the smaller nanoparticles may promote the delivery of DNA into cells through endocytosis and could improve transfection efficiency. In addition, PS-CaP could be used to transfect HEK 293 cells after one week of storage at 4°C with a lesser extent of efficiency loss compared with classical calcium phosphate, indicating that protamine sulfate may increase the stability of calcium phosphate nanoparticles. The cell viability inhibition assay indicated that both nanoparticles show similar low cell toxicity. Conclusion: PS-CaP can be used as a better nonviral transfection vector compared with classical calcium phosphate.
Evaluation of Hemodynamic Changes Due to Protamine Administration by Calcium Gluconate after Coronary Artery Bypass Grafting Surgery
M Janati,MT Moeinvaziri,J Kojuri,A Alipoor
Iranian Cardiovascular Research Journal , 2009,
Abstract: Background: Administration of Protamine sulfate for heparin neutralization after cardiopulmonary bypass may be associated with adverse reactions such as transient hypotension to cardiovascular collapse. Although catastrophic events are rare and occur only in 2.6% of cardiac surgeries, it is associated with adverse postoperative outcome. The aim of this study is to investigate whether bolus administration of calcium gluconate can minimize the adverse hemodynamic effects of protamine.Patients and Methods: This randomized clinical trial (RCT) prospective study was conducted between Feb. 2006 to Dec. 2008. The patients were randomly allocated into three groups including group A (42 patients) who received only protamine after weaning from cardiopulmonary, group B (44 patients) concomitantly treated with protamine sulfate and calcium gluconate, and group C (40 patients) receiving calcium gluconate 5 minutes before administration of protamine. Hemodynamic variables such as systolic and diastolic blood pressures, mean of arterial pressure, central venous pressure and heart rate were obtained 0, 2, 4, 6, 8 and 10 minutes after protamine administration from each group.Results: Systolic blood pressure in groups A (control) and C (calcium administration before protamine) 0,2,4,6,8 and 10 minutes after protamine administration initially decreased and increased subsequently (P=0.228). Also no statistically significant difference was found in diastolic blood pressure (DBP), mean arterial pressure (MAP), central venous pressure (CVP), and heart rate (HR) in 0,2,4,6,8, and 10 minutes in any of the three groups. Conclusion: In our study, hemodynamic changes in 10 minutes after protamine administration for heparin neutralization in patients with good left ventricular systolic function after coronary artery bypass grafting surgery were mild, and prophylactic calcium gluconate administration concurrent with or before protamine injection was not recommended.
Single Nucleotide Polymorphism Analysis of Protamine Genes in Infertile Men
Ahamad Salamian,Kamran Ghaedi,Shahnaz Razavi,Mahmud Tavalaee
International Journal of Fertility & Sterility , 2008,
Abstract: Background: Single nucleotide polymorphism (SNPs) are considered as one of the underlyingcauses of male infertility. Proper sperm chromatin packaging which involves replacement ofhistones with protamines has profound effect on male fertility. Over 20 SNPs have been reportedfor the protamine 1 and 2.Materials and Methods: The aim of this study was to evaluate the frequency of two previouslyreported SNPs using polymerase chain reaction (PCR)-restriction fragment length polymorphism(RFLP) approach in 35, 96 and 177 normal, oligozoospermic and azoospermic individuals. TheseSNPs are: 1. A base pair substitution (G) at position 197 instead of T in protamine type 1 Openreading frame (ORF) including untranslated region, which causes an Arg residue change to Serresidue in a highly conserved region. 2. cytidine nucleotide change to thymidine in position of 248of protamine type 2 ORF which caused a nonsense point mutation.Results: The two mentioned SNPs were not present in the studied population, thus concluding thatthese SNPs can not serves as molecular markers for male infertility diagnosis.Conclusion: The results of our study reveal that in a selected Iranian population, the SNP G197Tand C248T are completely absent and are not associated with male infertility and therefore theseSNPs may not represent a molecular marker for genetic diagnosis of male infertility.
Semen Parameters and Chromatin Packaging in Microsurgical Varicocelectomy Patients
Marziyeh Tavalaee,Homayon Abbasi,Mohammad Reza Deemeh
International Journal of Fertility & Sterility , 2012,
Abstract: Background: Varicocelectomy is considered as standard treatment for male infertility for clinicalvaricocele. The aim of this study is to address the effects of varicocelectomy on semen parameters,chromatin packaging, and pregnancy outcome.Materials and Methods: This retrospective study was carried out between June 2006 and February2011 on 145 infertile men with grade II or III varicocele. Microsurgical varicocelectomy wasperformed as part of patient management. Sperm count, motility, morphology, and chromatinpackaging were assessed with a Makler counting chamber, light microscopy, Papanicoulaou andchromomycin A3 (CMA3) staining, respectively. In addition, we assessed spontaneous clinicalpregnancy and miscarriage rates.Results: The percentages of spontaneous cumulative pregnancies post-surgery were 33.1% (3months), 42.06% (6 months), 46.2% (9 months), 48.9% (12 months), and 55.8% (after 12 months).Percentages of spontaneous cumulative miscarriage post-surgery were 2.46% (3 months), 4.93%(6 months), 4.93% (9 months), 6.17% (12 months), and 6.17 % (after 12 months). Both spermparameters improved and the percentage of sperm protamine deficiency decreased significantlyafter varicocelectomy.Conclusion: These results confirm that varicocelectomy improves sperm parameters and chromatinpackaging, thereby improving the chance of pregnancy. Positive aspects of this study include thelarge number of patients studied, duration of follow up, one surgeon who performed all of thesurgeries, and type of surgery (microsurgery). The spontaneous pregnancy results also suggest thatif pregnancy is not achieved within twelve months post-surgery, an alternative approach such asassisted reproductive technology (ART) treatment should be considered.
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