Respiratory syncytial virus (RSV) in Alaska Native children from the Yukon Kuskokwim (YK) Delta is associated with a hospitalization rate five times higher than that reported for the general US child population. The role of other viral respiratory pathogens has not been studied in this population. YK Delta children <3 years of age hospitalized with respiratory infections and same aged community control children were prospectively enrolled between October 2005 and September 2007. Polymerase chain reaction detection of viruses was performed on nasopharyngeal samples. Characteristics of hospitalized and asymptomatic control children were analyzed. From October 2005 to September 2007, 440 hospitalized and 425 control children were analyzed. Respiratory viruses were detected in 90% (395) of hospitalized children: 194 (44%) rhinovirus, 131 (30%) adenovirus, 102 (23%) RSV, 77 (18%) para influenza viruses (PIV), 66 (15%) human metapneumovirus (hMPV), 23 (5%) influenza, and 25 (6%) coronavirus. Fifty-two percent (221) of control children had a virus detected, most commonly rhinovirus (33%), and adenovirus (16%). RSV, PIV, hMPV, and influenza were significantly more common in hospitalized cases than control children, but rhinovirus, adenovirus, and coronavirus were not. RSV and hMPV were associated with higher severity of illness. In this study, RSV remains the most important virus associated with respiratory hospitalization, although hMPV and PIV were also common. RSV and hMPV were associated with more severe illness. Rhinovirus and adenovirus were detected in two-thirds of hospitalized children, but their frequent detection in control children made their role in respiratory hospitalization uncertain.

The human adenovirus 2 (Ad2) transforming region is located in the left 11.1% of the viral genome and encodes two early transcription units, ElA and E1B. Based on the amino acid sequence deduced from the Ad2 E1B DNA sequence (Gingeras et al., J. Biol. Chem. 257:13475-13491, 1982), we have prepared antibodies against synthetic peptides, 8 to 16 amino acids in length, encoded at the NH2 and COOH termini of the major ElB-19K and ElB-53K tumor antigens. The antipeptide antibodies immunoprecipitated the targeted ElB-19K or ElB-53K tumor antigens from extracts of Ad2-infected cells. The specificity of the antipeptide antibody recognition of the EIB tumor antigens was confirmed by peptide competition studies. Antipeptide antibodies directed to the NH2 and COOH termini immunoprecipitated the ElB-19K and ElB-53K tumor antigens from two Ad2-transformed rat cell lines, F17 and F4, providing evidence that identical tumor antigens are synthesized in Ad2-infected and Ad2-transformed cells. These results show that the ElB-19K and ElB-53K T antigens are not processed proteolytically at either the NH2 or COOH terminus. Our data provide strong evidence at the protein level that the ElB-19K and ElB-53K tumor antigens partially overlap in DNA sequence, with the ElB-19K initiating translation at the first ATG at nucleotide 1711 in translation reading frame 1 and the ElB-53K tumor antigen initiating translation at the second ATG at nucleotide 2016 in reading frame 3. This confirms the results of others on the N-terminal amino acid sequence of ElB-19K and theoretical deductions based on the DNA sequence. Our findings prove that the large ElB-53K T antigen initiates translation at the second ATG at nucleotide 2016 and not at equally plausible initiation codons located farther downstream at nucleotides 2202 and 2235. Thus, the ElB-53K T antigen is another example of a protein which initiates translation at an internal ATG rather than at the 5'-proximal ATG. During the early stages of adenovirus 2 (Ad2) productive infection, seven regions of the viral genome are transcribed, early region 1A (ElA), E1B, E2A, E2B, E3, E4, and late region 1 (Li) (see reference 45 for a recent review). EIA, located at map position 1.3 to 4.5, and E1B, located at map position 4.6 to 11.1, are of special interest because they encode the information for Ad2-induced cell transformation. Much is known about El of Ad2 and the closely related AdS (both group C). The DNA encoding the transforming genes has been sequenced (7, 16, 57). The major El-coded mRNAs and proteins have been identified by cell-free translation and by immunoprecipitation with antisera against Ad-transformed cells (Ad tumor sera). Cell-free translation studies with Ad2 and AdS El mRNAs have detected about eight proteins that are encoded by ElA and E1B (7, 13, 17, 23, 24, 35, 37, 38). As determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, two early proteins of apparent Mr 41,000 to 53,000 are translated from the ElA-13S mRNA, two early proteins of apparent Mr 35,000 to 4...

Dialyzed extracts of the gentamicin producer Micromonospora purpurea catalyze reactions which represent transaminations proposed for 2-deoxystreptamine biosynthesis. To determine whether these transaminations were catalyzed by a single aminotransferase or by multiple enzymes, we purified and characterized an L-glutamine:keto-scyllo-inositol aminotransferase from M. purpurea. This enzyme was purified 130-to 150-fold from late-log-phase mycelia of both wild-type M. purpurea and a 2-deoxystreptamine-less idiotroph. The cofactor pyridoxal phosphate was found to be tightly bound to the enzyme, and spectral analysis demonstrated its participation in the transamination reactions of this enzyme. The major physiological amino donor for the enzyme appears to be L-glutamine; the keto acid product derived from glutamine was characterized as 2-ketoglutaramate, indicating that the alpha amino group of glutamine participates in the transamination. We found that crude extracts contained omega-amidase activity, which may render transaminations with glutamine irreversible in vivo. The substrate specificity of the aminotransferase was shown to be restricted to deoxycyclitols, monoaminocyclitols, and diaminocyclitols, glutamine, and 2-ketoglutaramate, which contrasts with the broader substrate specificity of mammalian glutamine aminotransferase. The appearance of the enzyme in late-log phase, coupled with its narrow substrate specificity, indicates that it participates predominantly in 2-deoxystreptamine biosynthesis rather than in general metabolism. The enzyme catalyzes reactions which represent both transamination steps of 2-deoxystreptamine biosynthesis. Although copurification of two aminotransferases cannot be ruled out, our data are consistent with the participation of a single aminotransferase in the formation of both amino groups of 2-deoxystreptamine during biosynthesis by M. purpurea. We propose that this aminotransferase participates in a key initial step in the biosynthesis of most aminocyclitol antibiotics.2-Deoxystreptamine is a component of numerous clinically important antibiotics such as gentamicin, neomycin, tobramycin, amikacin, and hygromycin, while streptamine is known to occur naturally in place of 2-deoxystreptamine in at least one member (SS-56C) of the destomycin group (for reviews, see references 19 and 21). A pathway for 2-deoxystreptamine biosynthesis has been proposed (Fig. 1) on the basis of mutasynthesis and isotopic competition studies (9, 13) and in vitro enzymatic studies (5). The two aminotransferase steps of this pathway may be analogous to the two transaminations which occur in the biosynthesis of the streptidine moiety of streptomycin (27). These reactions in streptidine biosynthesis are catalyzed by separate enzymes; L-glutamine:keto-scyllo-inositol aminotransferase (glutamine-scyllo-inosose aminotransferase; EC 2.6.1.50) catalyzes the first transamination reaction (26). This enzymatic activity has also been detected in extracts of actinomycetes that produce bluensomycin (26) and the 2-deoxys...

COOH (peptide 204), targeted to the common C-terminus of human adenovirus 12 (Adl2) tumor antigens encoded by the ElA 13S mRNA and 12S mRNA, has been synthesized. Antibody prepared in rabbits against peptide 204 immunoprecipitated two proteins of apparent Mr 47,000 and 45,000 from extracts of [35S]methionine-labeled Adl2-early infected KB cells and a 47,000 protein from extracts of the Adl2-transformed hamster cell line, HE C19. Immunoprecipitation analysis of infected and transformed cells labeled with 32P, showed that both major Adl2 EIA T antigens are phosphoproteins. Immunofluorescence microscopy of Adl2-early infected KB cells with antipeptide antibody showed the site of ElA protein concentration to be predominantly nuclear. ElA proteins were detected by imtnunofluorescence at 4 to 6 h postinfection and continued to increase until at least 18 h postinfection. Antipeptide 204 antibody was used to analyze the proteins synthesized in Escherichia coli cells transformed by plasmids containing cDNA copies of the Adl2 EIA 13S mRNA or 12S mRNA under the control of the tac promoter (D.

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