ABSTRACT
BACKGROUND: Eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) recommendations are frequently stated at 500 mg/d; however, adherence to these recommendations would result in a large global commercial EPA/DHA production deficit. Previously, our laboratory demonstrated that acute DHA intake in rats can increase the capacity for synthesis-secretion of n-3 (ω-3) polyunsaturated fatty acids (PUFAs). OBJECTIVE: We aimed to investigate the utility of a dietary DHA cycling strategy that employs 2 wk of repeated DHA feeding for a total of 3 cycles over 12 wk. METHODS: Male Long-Evans rats were fed a 10% fat diet by weight comprised of either 1) a 2-wk, 2% α-linolenic acid (ALA, DHA-ALA group 18:3n-3) diet followed by a 2-wk, 2% DHA + 2% ALA diet over 3 consecutive 4-wk periods ("DHA cycling," DHA-ALA group); 2) a 2% DHA + 2% ALA diet (DHA group) for 12 wk; or 3) a 2% ALA-only diet (ALA group) for 12 wk. At 15 wk old, blood and tissue fatty acid concentrations and liver mRNA expression and 13C-DHA natural abundances were determined. RESULTS: DHA concentrations in plasma, erythrocytes, and whole blood between the DHA-ALA group and the DHA groups were not different (P ≥ 0.05), but were 72-110% higher (P < 0.05) than in the ALA group. Similarly, DHA concentrations in liver, heart, adipose, and brain were not different (P ≥ 0.05) between the DHA-fed groups, but were at least 62%, 72%, 320%, and 68% higher (P < 0.05) than in the ALA group in liver, heart, adipose, and skeletal muscle, respectively. Compound-specific isotope analysis indicated that 310% more liver DHA in the DHA-ALA group compared with the DHA group is derived from dietary ALA, and this was accompanied by a 123% and 93% higher expression of elongation of very long-chain (Elovl)2 and Elovl5, respectively, in the DHA-ALA group compared with the ALA group. CONCLUSIONS: DHA cycling requires half the dietary DHA while achieving equal blood and tissue DHA concentrations in rats. Implementation of such dietary strategies in humans could reduce the gap between global dietary n-3 PUFA recommendations and commercial production.
Subject(s)
Docosahexaenoic Acids/pharmacology , alpha-Linolenic Acid/metabolism , Adipose Tissue/chemistry , Animals , Brain Chemistry , Docosahexaenoic Acids/administration & dosage , Dose-Response Relationship, Drug , Drug Administration Schedule , Erythrocytes , Fatty Acids/chemistry , Fatty Acids/metabolism , Mumps Vaccine/chemistry , Muscle, Skeletal/chemistry , Myocardium/chemistry , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Long-EvansABSTRACT
[In the present work virus particles of live mumps virus vaccine widely used for vaccination in Russia have been detected and visualized by atomic force microscopy. For quantitative estimation of the number of observed virus particles the special method has been proposed. The presence of protein component of the virus in vaccine was tested by ELISA and dot-blot analysis. Using quantitative real-time PCR assay the number of copies of viral RNA was estimated. The results of quantitative estimation obtained by real-time PCR corresponded with atomic force microscopy data.
Subject(s)
Mumps Vaccine/chemistry , Mumps Vaccine/genetics , Mumps Vaccine/immunology , Mumps virus/genetics , Mumps virus/immunology , Mumps virus/ultrastructure , Enzyme-Linked Immunosorbent Assay/methods , Humans , Microscopy, Atomic Force/methods , RNA, Viral/genetics , Reverse Transcriptase Polymerase Chain Reaction/methodsABSTRACT
A point mutation from guanine (G) to adenine (A) at nucleotide position 1081 in the hemagglutinin-neuraminidase (HN) gene has been associated with neurovirulence of Urabe AM9 mumps virus vaccine. This mutation corresponds to a glutamic acid (E) to lysine (K) change at position 335 in the HN glycoprotein. We have experimentally demonstrated that two variants of Urabe AM9 strain (HN-A1081 and HN-G1081) differ in neurotropism, sialic acidbinding affinity and neuraminidase activity. In the present study, we performed a structure-function analysis of that amino acid substitution; the structures of HN protein of both Urabe AM9 strain variants were predicted. Based on our analysis, the E/K mutation changes the protein surface properties and to a lesser extent their conformations, which in turn reflects in activity changes. Our modeling results suggest that this E/K interchange does not affect the structure of the sialic acid binding motif; however, the electrostatic surface differs drastically due to an exposed short alpha helix. Consequently, this mutation may affect the accessibility of HN to substrates and membrane receptors of the host cells. Our findings appear to explain the observed differences in neurotropism of these vaccine strains.
Subject(s)
Genetic Variation/genetics , HN Protein/genetics , Mumps Vaccine/genetics , Mumps virus/genetics , Amino Acid Substitution/genetics , Animals , Cell Line, Tumor , Chlorocebus aethiops , Genetic Variation/immunology , HN Protein/chemistry , Humans , Mumps Vaccine/chemistry , Mumps virus/immunology , Point Mutation , Structure-Activity Relationship , Vero CellsABSTRACT
A point mutation from guanine (G) to adenine (A) at nucleotide position 1081 in the hemagglutinin-neuraminidase (HN) gene has been associated with neurovirulence of Urabe AM9 mumps virus vaccine. This mutation corresponds to a glutamic acid (E) to lysine (K) change at position 335 in the HN glycoprotein. We have experimentally demonstrated that two variants of Urabe AM9 strain (HN-A1081 and HN-G1081) differ in neurotropism, sialic acidbinding affinity and neuraminidase activity. In the present study, we performed a structure-function analysis of that amino acid substitution; the structures of HN protein of both Urabe AM9 strain variants were predicted. Based on our analysis, the E/K mutation changes the protein surface properties and to a lesser extent their conformations, which in turn reflects in activity changes. Our modeling results suggest that this E/K interchange does not affect the structure of the sialic acid binding motif; however, the electrostatic surface differs drastically due to an exposed short alpha helix. Consequently, this mutation may affect the accessibility of HN to substrates and membrane receptors of the host cells. Our findings appear to explain the observed differences in neurotropism of these vaccine strains.
Subject(s)
Animals , Humans , Genetic Variation/genetics , HN Protein/genetics , Mumps Vaccine/genetics , Mumps virus/genetics , Amino Acid Substitution/genetics , Cell Line, Tumor , Chlorocebus aethiops , Genetic Variation/immunology , HN Protein/chemistry , Mumps Vaccine/chemistry , Mumps virus/immunology , Point Mutation , Structure-Activity Relationship , Vero CellsABSTRACT
A novel measles-mumps-rubella vaccine (Priorix) has been marketed by SmithKline Beecham. It contains live attenuated virus with measles and mumps strains slightly different from those present in MMR VAX (Pasteur Merieux MSD). The indications and contraindications are similar for both vaccines. Immunogenicity is also equivalent as well as general reactogenicity. By contrast local symptoms were reported significantly less frequently after Priorix.
Subject(s)
Measles Vaccine , Mumps Vaccine , Rubella Vaccine , Vaccines, Combined , Adjuvants, Immunologic , Child , Child, Preschool , Contraindications , Humans , Immunization Schedule , Infant , Measles Vaccine/adverse effects , Measles Vaccine/chemistry , Measles Vaccine/immunology , Measles-Mumps-Rubella Vaccine , Mumps Vaccine/adverse effects , Mumps Vaccine/chemistry , Mumps Vaccine/immunology , Rubella Vaccine/adverse effects , Rubella Vaccine/chemistry , Rubella Vaccine/immunology , Vaccination , Vaccines, Combined/adverse effects , Vaccines, Combined/chemistry , Vaccines, Combined/immunologyABSTRACT
The reason for the high incidence of vaccine-associated meningitis due to the Urabe AM9 vaccine was assessed by comparing the nucleotide (nt) sequence of the hemagglutinin-neuraminidase (HN) gene from vaccine virus to those of viruses isolated from persons with postvaccination meningitis. A G1081--> A nt substitution that was predicted to result in a Glu335--> Lys reversion in the HN protein was detected between Urabe AM9 (G) and postvaccine meningitis mumps virus isolates (A). Further analysis showed that the Urabe AM9 vaccine was a mixture of viruses with wild type (A) and variant (G) nt at position 1081. Urabe AM9 vaccinees who developed meningitis or parotitis possessed predominantly A (98%-100%) at nt 1081, indicating strong selection of the wild type (A) form relative to the variant (G) form. Mumps virus homogeneous for the variant Glu335 form of the HN gene may be safer than the original Urabe AM9 vaccine.
Subject(s)
HN Protein/chemistry , Mumps Vaccine/chemistry , Mumps virus/chemistry , Animals , Base Sequence , Chlorocebus aethiops , Genetic Variation , HN Protein/genetics , Humans , Meningitis, Viral/etiology , Molecular Sequence Data , Mumps Vaccine/adverse effects , Mumps virus/genetics , Sequence Analysis, DNA , Vero Cells , VirulenceABSTRACT
We examined live virus vaccines against measles, mumps, and rubella for the presence of pestivirus RNA or of pestiviruses by reverse transcription PCR. Pestivirus RNA was detected in two measles-mumps-rubella combined vaccines and in two monovalent vaccines against mumps and rubella. Nucleotide sequence analysis of the PCR products indicated that a modified live vaccine strain used for immunization of cattle against bovine viral diarrhea is not responsible for the contamination of the vaccines.
