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Consequences of congenital hearing loss - a longterm view.

Responsibility for detection of hearing loss at the earliest possible age rests on the shoulders of the medical profession. Early detection and presentation of language through all available sensory channels, most notably the visual, are essential since deaf children of hearing parents, the vast majority of the deaf, characteristically remain grossly deficient linguistically despite years of special schooling. The deaf children of deaf parents, who are not as deprived linguistically since they learn a manual language at the normal age of language acquisition, tend to be less severely handicapped than the deaf children of hearing parents. Deafness is a hidden handicap in infancy although lack of vestibular function may delay motor milestones like sitting and walking but does not constitute a serious problem after they are achieved. The average 18 year old deaf student achieves a reading level comparable to that of a hearing fourth grader and the mathematical skills of a sixth grader. He is also deficient in science, social studies, and general information despite his average scores on nonverbal intelligence tests. Although many deaf students drop out of school before graduation and few go on to higher education, congenitally deaf adults are usually self-supporting and independent. Eighty per cent of the deaf marry hearing impaired persons. The deaf are greatly assisted by the cohesive deaf community existing in every sizable city which provides a social, recreational, and economic framework to their lives. As a result, delinquency and psychiatric illness are not higher among the deaf than the hearing, despite the difficulties they have dealing with the complexities of the hearing society.

Adolescent↗

Sequencing and analysis of the large virulence plasmid pLVPK of Klebsiella pneumoniae CG43.

We have determined the entire DNA sequence of pLVPK, which is a 219-kb virulence plasmid harbored in a bacteremic isolate of Klebsiella pneumoniae. A total of 251 open reading frames (ORFs) were annotated, of which 37% have homologous genes of known function, 31% match the hypothetical genes in the GenBank database, and the remaining 32% are novel sequences. The obvious virulence-associated genes carried by the plasmid are the capsular polysaccharide synthesis regulator rmpA and its homolog rmpA2, and multiple iron-acquisition systems, including iucABCDiutA and iroBCDN siderophore gene clusters, Mesorhizobium loti fepBC ABC-type transporter, and Escherichia coli fecIRA, which encodes a Fur-dependent regulatory system for iron uptake. In addition, several gene clusters homologous with copper, silver, lead, and tellurite resistance genes of other bacteria were also identified. Identification of a replication origin consisting of a repA gene lying in between two sets of iterons suggests that the replication of pLVPK is iteron-controlled and the iterons are the binding sites for the repA to initiate replication and maintain copy number of the plasmid. Genes homologous with E. coli sopA/sopB and parA/parB with nearby direct DNA repeats were also identified indicating the presence of an F plasmid-like partitioning system. Finally, the presence of 13 insertion sequences located mostly at the boundaries of the aforementioned gene clusters suggests that pLVPK was derived from a sequential assembly of various horizontally acquired DNA fragments.

Amino Acid Sequence↗

Role of speech therapy and sign language prior to cochlear implantation.

The speech therapist should assess and develop the communication skills of the deaf child prior and after a cochlear implantation. The goals are first the acquisition of a useful communication system by the deaf child (for avoiding a cognitive stimulation lack) and parallely the acquisition of good oral language competencies. The different modes of communication useable in the profoundly deaf education are presented. Exclusive sign language use is not indicated in implanted children. It is well demonstrated that an exclusive oral communication is associated with an insufficient knowledge of the oral language. The total communication (combination of listening, speech reading, signing with or without finger spelling) is a defective communication mode with a bad representation of the morpho-syntactic and the phonologic aspects of the oral language. The best choice seems to be the simultaneously use of a spoken language and a manually coding of their phonological structures. The limits of this alternative are discussed.

Child↗

Speech, language and literacy skills 3 years later: a follow-up study of early phonological and metaphonological intervention.

BACKGROUND: Three years before the present study, 19 preschool children participated in a phonological and metaphonological intervention programme. The phonological intervention programme was based on non-linear phonological analyses. The metaphonological intervention programme included both rhyming and alliteration tasks and was directly targeted during the last section of the programme. All children made significant gains in phonology, and many in metaphonology. However, the literature notes a tendency for children with history of early speech or language impairments to have difficulty acquiring literacy skills or to show residual speech impairment. The participants of the 1998 study were therefore considered at risk for continuing speech impairment and/or academic performance. AIMS: The primary objectives were to document the children's later speech, language and literacy skills, and to determine potential relationships between previous and concurrent child factors. METHODS & PROCEDURES: Twelve children from the original cohort (aged 6;1-8;5) received a comprehensive battery of speech, language, cognitive and academic tasks 3 years after the original study. Data were evaluated both concurrently and in terms of the children's preschool performance. OUTCOMES & RESULTS: Five of the children had residual phonological impairment. Only two children showed below average reading (decoding and comprehension), although five also showed below average spelling performance. Children with limitations in verbal memory, language production and metaphonology at the follow-up point were more likely to show delays in literacy. Language production skills at the follow-up point were most strongly correlated with literacy development. In terms of the early study, the strongest predictor for literacy development was performance on metaphonology tasks at the end of the early intervention study. The strongest predictor for ongoing speech impairment was phonological skill at the end of the early study. CONCLUSIONS: Results suggest that early phonological and metaphonological intervention can promote normalization of speech development and normal acquisition of literacy skills for at least some children with a history of severe phonological impairment. In the earlier study, the component structure of words (onsets, rhymes, codas) was emphasized through both metaphonological and non-linear phonological intervention. Risk for literacy and ongoing speech impairment can be reduced through early intervention that draws attention to the structure of words.

Child↗

The complete nucleotide sequence of phi CTX, a cytotoxin-converting phage of Pseudomonas aeruginosa: implications for phage evolution and horizontal gene transfer via bacteriophages.

phi CTX is a cytotoxin-converting phage isolated from Pseudomonas aeruginosa. In this study, we determined the complete nucleotide sequence of the phi CTX phage genome. The precise genome size was 35,538 bp with 21 base 5'-extruding cohesive ends. Forty-seven open reading frames (ORFs) were identified on the phi CTX genome, including two previously identified genes, ctx and int. Among them, 15 gene products were identified in the phage particle by protein microsequencing. The most striking feature of the phi CTX genome was an extensive homology with the coliphage P2 and P2-related phages; more than half of the ORFs (25 ORFs) had marked homology to P2 genes with 28.9-65.8% identity. The gene arrangement on the genome was also highly conserved for the two phages, although the G + C content and codon usage of most phi CTX genes were similar to those of the host P. aeruginosa chromosome. In addition, phi CTX was found to share several common features with P2, including the morphology, non-inducibility, use of lipopolysaccharide core oligosaccharide as receptor and Ca(2+)-dependent receptor binding. These findings indicate that phi CTX is a P2-like phage well adapted to P. aeruginosa, and provide clear evidence of the intergeneric spread and evolution of bacteriophages. Furthermore, comparative analysis of genome structures of phi CTX, P2 and other P2 relatives revealed the presence of several hot-spots where foreign DNAs, including the cytotoxin gene, were inserted. They appear to be deeply concerned in the acquisition of various genes that are horizontally transferred by bacteriophage infection.

Amino Acid Sequence↗

Language processing modulated by literacy: a network analysis of verbal repetition in literate and illiterate subjects.

Previous behavioral and functional neuroimaging data indicate that certain aspects of phonological processing may not be acquired spontaneously, but are modulated by learning an alphabetic written language, that is, learning to read and write. It appears that learning an alphabetic written language modifies the auditory-verbal (spoken) language processing competence in a nontrivial way. We have previously suggested, based on behavioral and functional neuroimaging data, that auditory-verbal and written language interact not only during certain language tasks, but that learning and developing alphabetic written language capacities significantly modulates the spoken language system. Specifically, the acquisition of alphabetic orthographic knowledge has a modulatory influence on sublexical phonological processing and the awareness of sublexical phonological structure. We have suggested that developing an orthographic representation system for an alphabetic written language, and integrating a phoneme-grapheme correspondence with an existing infrastructure for auditory-verbal language processing, will result in a modified language network. Specifically, we suggest that the parallel interactive processing characteristics of the underlying language-processing brain network differ in literate and illiterate subjects. Therefore, the pattern of interactions between the regions of a suitably defined large-scale functional-anatomical network for language processing will differ between literate and illiterate subjects during certain language tasks. In order to investigate this hypothesis further, we analyzed the observed covariance structure in a PET data set from a simple auditory-verbal repetition paradigm in literate and illiterate subjects, with a network approach based on structural equation modeling (SEM). Based on a simple network model for language processing, the results of the present network analysis indicate that the network interactions during word and pseudoword repetition in the illiterate group differ, while there were no significant differences in the literate group. The differences between the two tasks in the illiterate group may reflect differences in attentional modulation of the language network, executive aspects of verbal working memory and the articulatory organization of verbal output. There were no significant differences between the literate and illiterate group during word repetition. In contrast, the network interactions differed between the literate and illiterate group during pseudoword repetition. In addition to differences similar to those observed in the illiterate group between word and pseudoword repetition, there were differences related to the interactions of the phonological loop between the groups. In particular, these differences related to the interaction between Broca's area and the inferior parietal cortex as well as the posterior-midinsula bridge between Wernicke's and Broca's area. In conclusion, the results of this network analysis are consistent with our previously presented results and support the hypothesis that learning to read and write during childhood influences the functional architecture of the adult human brain. In particular, the basic auditory-verbal language network in the human brain is modified as a consequence of acquiring orthographic language skills.

Adult↗

Tomato leaf curl virus from Bangalore (ToLCV-Ban4): sequence comparison with Indian ToLCV isolates, detection in plants and insects, and vector relationships.

Tomato leaf curl virus (ToLCV) is a whitefly (Bemisia tabaci) transmitted geminivirus (family Geminiviridae, genus Begomovirus) causing a destructive disease of tomato in many regions of India, East Asia and Australia. While ToLCV isolates from Australia and Taiwan have a single genomic component (designated DNA-A), those from Northern India have two components (DNA-A and DNA-B). The ToLCV isolates from Southern India (Bangalore) previously cloned seem to have a DNA-A-like monopartite genome. We have used degenerate DNA-A-specific PCR primers to clone the genome of a ToLCV isolate (named ToLCV-Ban4) from field-infected tomato plants growing in Bangalore, India, in 1997. Degenerate DNA-B-specific PCR primers have not allowed to amplify a putative DNA-B from infected tomato, at the time when DNA-B fragments were amplified from plants infected by known bipartite begomoviruses. The full-length 2759 nucleotide-long DNA-A-like viral genome was sequenced. Similarly to other monopartite ToLCV and TYLCV isolates, ToLCV-Ban4 contains six open reading frames, two on the virion strand and four on the complementary strand. Sequence comparisons indicated that ToLCV-Ban4 is similar to the other three isolates from Bangalore previously sequenced, and is closely related to ToLCV-Ban2 (approximately 91% nucleotide sequence identity). Phylogenetic analysis showed that the ToLCV isolates from Bangalore constitute a group of viruses separated from those of Northern India. ToLCV-Ban4 was detected in tomato and in its whitefly vector Bemisia tabaci by one or by a combination of ELISA, Southern blot hybridization and PCR. Parameters of virus acquisition, retention and transmission by the whitefly vector were investigated in the laboratory. Single whiteflies were able to acquire ToLCV-Ban4 from infected tomato and to transmit the virus to tomato test plants, but five insects were necessary to achieve 100% transmission. Minimum acquisition access and inoculation access periods were 10 min and 20 min, respectively. A latent period of 6 h was required for B. tabaci to efficiently infect tomato test plants. Following a 24 h acquisition access period the insect retained its ability to infect tomato test plants for 12 days, but not for its entire life. In one insect/one plant inoculation tests, female whiteflies were more efficient (approximately 95%) than males (approximately 25%) in transmitting the virus.

Animals↗

Acquisition of apparent DNA slippage structures during extensive evolutionary divergence of pcaD and catD genes encoding identical catalytic activities in Acinetobacter calcoaceticus.

The pca operon from the Gram- bacterium Acinetobacter calcoaceticus encodes all of the enzymes required for catabolism of protocatechuate to common intermediary metabolites. This report presents the 2754-nucleotide (nt) sequence of a HindIII restriction fragment containing pcaD, the 801-bp gene encoding beta-ketoadipate enol-lactone hydrolase I. The deduced primary structure of A. calcoaceticus PcaD shares 44% amino acid (aa) sequence identity with the aligned primary structure of CatD (beta-ketoadipate enol-lactone hydrolase II) from the same organism, and the overall nt sequence identity of the two genes is 51.8%. In the 56% of the genes where selection for identical aa residues was not imposed, pcaD and catD have diverged so extensively that nt sequence identity of the aligned segments is only 28.2%; the G+C contents of these segments from the respective genes differ by 8%. Conserved within the aligned PcaD and CatD aa sequences is a Ser residue corresponding to the nucleophile within the alpha/beta-fold of many hydrolytic enzymes. In this region of primary structure, PcaD and CatD appear to have maintained some different aa sequences derived from a common ancestor. Conservation of the different aa sequences during extreme evolutionary divergence suggests that separate segments of primary structure, conserved within either PcaD or CatD, may be functionally incompatible within recombinant enzymes. Consequently, selection for avoidance of genetic exchange between pcaD and catD could account for the thorough nt substitution in regions where identical aa were not selected. Sequence repetitions within pcaD suggest that the multiple mutations required for its extensive divergence from catD were achieved in part by acquisition of a complex DNA slippage structure.

Acinetobacter calcoaceticus↗

Are horizontal transfers involved in the evolution of the Streptococcus thermophilus exopolysaccharide synthesis loci?

A 32.5kb variable locus of the Streptococcus thermophilus CNRZ368 chromosome, the eps locus, contains 25 ORF and seven insertion sequences (IS). The putative products of 17 ORF are related to proteins involved in the synthesis of polysaccharides in various bacteria. The two distal regions and a small central region of the eps locus are constant and present in all or almost all of the S. thermophilus strains tested. The other regions are variable and present in only some S. thermophilus strains tested, particularly in the closely related strains CNRZ368 and A054. A 13.6kb variable region of the eps locus of S. thermophilus CNRZ368 contains two ORF that are almost identical to epsL and orfY of the eps locus of Lactococcus lactis NIZOB40 and seven IS belonging to four different families, ISS1, IS981, IS1193 and IS1194. Five of these sequences were probably acquired by horizontal transfer from L. lactis (Bourgoin, F., et al., 1996. Gene 178, 15-23). Three probes of this 13.6kb region hybridized with the DNA of several L. lactis strains tested. A specific probe for another sequence within the S. thermophilus eps locus, epsF, hybridized with the DNA of one of the L. lactis strains tested. Sequence comparisons also suggest that five ORF of the eps locus have a mosaic structure and probably result from recombinations between sequences that are 10 to 50% divergent. The chimeric structure of the eps locus suggests a very complex evolution. This evolution probably involves both the acquisition of the 13.6kb region from L. lactis by horizontal transfer and exchanges within the S. thermophilus species.

Chromosome Mapping↗

Fourier temporal interpolation improves electrocardiograph-gated myocardial perfusion SPECT.

UNLABELLED: The optimal temporal sampling rate in electrocardiograph-gated myocardial SPECT is questionable: low rates, typically 8 frames per cardiac beat (8fr/cb), favor image quality, whereas high rates, typically 16 frames per cardiac beat (16fr/cb), favor the accuracy of left ventricular (LV) functional parameters. We examined whether Fourier temporal interpolation (FTI) from 8fr/cb to 16fr/cb can combine the advantages of low and high rates. METHODS: In 34 patients imaged after stress injection of (99m)Tc-sestamibi, 4 sets of reconstructed gated slices were compared: a raw 16fr/cb acquisition (R16), a raw 8fr/cb acquisition (R8), a 16fr/cb set obtained by FTI of 8fr/cb projections (IP), and a 16fr/cb set obtained by FTI of 8fr/cb reconstructed slices (IS). LV ejection fraction (LVEF), end-diastolic volume (EDV), and end-systolic volume (ESV) obtained from the final LV volume curve were compared for the 4 datasets. Deviation of the whole LV volume curve was quantified for IP and IS with respect to R16. Image quality was evaluated by consensus reading of end-diastolic slices of the 4 sets. For R16, IP, and IS, cine display fluidity was quantified by a roughness index calculated from the LV volume curve. RESULTS: No differences in EDVs or ESVs were found among R16, IP, and IS, whereas R8 gave smaller EDVs and larger ESVs. LVEF was lower with R8, IP, and IS than with R16: -3.9%, -1.2%, and -1.3%, respectively. The LV volume curve was closer to R16 with IP than with IS. Image quality was better with IP and IS than with R8 and better with R8 than with R16. Cine display fluidity was better with IP than with R16 and better with R16 than with IS. CONCLUSION: FTI improved image quality not only over that provided by R16 but even over that provided by R8. The sole worsened LV functional parameter was LVEF, which was slightly underestimated with respect to that estimated by R16. Of the 2 FTI variants, IP was superior to IS for cine display fluidity and accuracy of the LV volume curve with respect to the data obtained with R16. Therefore, FTI to 16fr/cb performed before reconstruction on a pixel-by-pixel basis on 8fr/cb projections improves image quality and cine display fluidity over those of both R8 and R16 acquisitions at the sole cost of a 1% underestimation of LVEF.

Algorithms↗

Cloning, nucleotide sequence, and mutagenesis of a gene (irpA) involved in iron-deficient growth of the cyanobacterium Synechococcus sp. strain PCC7942.

We describe the cloning and sequencing of a gene from the cyanobacterium Synechococcus sp. strain PCC7942, designated irpA (iron-regulated protein A), that encodes for a protein involved in iron acquisition or storage. Polyclonal antibodies raised against proteins which accumulate during iron-deficient growth were used as probes to isolate immunopositive clones from a lambda gt11 genomic expression library. The clone, designated lambda gtAN26, carried a 1.7-kilobase (kb) chromosomal DNA insert and was detected by cross-reactivity with antibody against a 36-kilodalton protein. It was possible to map a 20-kb portion of the chromosome with various DNA probes from lambda gt11 and lambda EMBL-3 clones, and Southern blot analysis revealed that the irpA gene was present in a single copy and localized within a 1.7-kb PstI fragment. DNA sequencing revealed an open reading frame of 1,068 nucleotides capable of encoding 356 amino acids which yields a protein with a molecular weight of 38,584. The hydropathy profile of the polypeptide indicated a putative N-terminal signal sequence of 44 amino acid residues. IrpA is a cytoplasmic membrane protein as determined by biochemistry and electron microscopy immunocytochemistry. The upstream region of the irpA gene contained a consensus sequence similar to the aerobactin operator in Escherichia coli. This fact, plus a mutant with a mutation in irpA that is unable to grow under iron-deficient conditions, led us to suggest that irpA is regulated by iron and that the gene product is involved in iron acquisition or storage.

Amino Acid Sequence↗

Effects of methylphenidate on learning a 'beginning reading vocabulary' by normal adults.

The effects of methylphenidate on reading was studied by having nine adults learn a beginning vocabulary of 96 Chinese characters under placebo and 5 mg and 10 mg methylphenidate by two teaching methods which differed in the rate at which new items were introduced for practice. Using lists composed of 12 characters and their English equivalents, all items were presented prior to the start of practice trials (simultaneous method), or each item was introduced only as the subject practiced items previously presented (progressive method). Learning performance was analyzed in terms of total errors and errors made at two criterion stages in list acquisition. Overall performance was significantly facilitated by the low (5 mg) dose or methylphenidate, but only with the simultaneous method. Significant drug effects were also obtained at criterion stages which were associated with high error rates. It was concluded that drug effects are likely to be most pronounced in difficult learning situations and that dosage appears to be an important variable for consideration in further studies.

Adult↗

Virion association of IE62, the varicella-zoster virus (VZV) major transcriptional regulatory protein, requires expression of the VZV open reading frame 66 protein kinase.

IE62, the major transcriptional regulatory protein encoded by varicella-zoster virus (VZV), is associated with the tegument of gradient-purified virions. Here, we show that most, if not all, of the association requires the expression of open reading frame 66 (ORF66), a protein kinase. The association of IE62 with wild-type VZV virions was confirmed using immunoelectron microscopy with IE62-specific antibodies, which reacted with virions in ultrathin sections of VZV-infected cells. Fractionated purified virions from cells infected with recombinant VZV ROka contained substantial levels of the 175-kDa virion IE62 protein and also contained the ORF66 protein. However, virions from cells infected with recombinant VZV ROka66S, in which ORF66 is disrupted, lacked not only the ORF66 protein but also most of the virion 175-kDa IE62 polypeptide. The virion-associated protein kinase activity was still present in ROka66S virions, although the 175-kDa protein substrate for the virion kinase was absent, implying that the virion protein kinase is encoded by genes other than ORF66. The very low levels of IE62 in ROka66S virions indicate that ORF66 protein mediates the redistribution of IE62 to sites of tegument assembly. IE62 was resolved into several species from VZV-infected cells which showed mobility differences between ROka and ROka66S, and a specific form of IE62 was detected in ROka virions. These results are consistent with a role for the ORF66-mediated phosphorylation of IE62 that results in cytoplasmic distribution of the regulatory protein for tegument inclusion. They support a model in which VZV tegument acquisition occurs in the cytoplasm. As such, two unusual features of VZV IE62, namely, its virion inclusion and its phosphorylation and nuclear exclusion by the ORF66 protein kinase, are functionally linked.

Herpesvirus 3, Human↗

Oviductin, the oviductal protease that mediates gamete interaction by affecting the vitelline coat in Bufo japonicus: its molecular cloning and analyses of expression and posttranslational activation.

Previous studies indicated that the acquisition of egg fertilizability during transit through the pars recta portion of the oviduct in Bufo japonicus is accompanied by hydrolytic conversion of the vitelline coat 40- to 52-kDa components to 39-kDa components induced by a 66-kDa serine protease, "oviductin." In this study, we cloned a 3028-bp cDNA that contained an open reading frame encoding 974 amino acids with a calculated molecular mass of 107.6 kDa, including two protease domains and three repeats of CUB domains. Sequence analysis indicated that the catalytically active 66-kDa protein comprised an N-terminally located oviductin protease and two CUB domains. The oviductin gene was transcribed as a part of 6-kb mRNA that was expressed specifically in the cells lining the bottom of epithelial folds in the oviductal pars recta, and this expression was highly accelerated when the pars recta fragments were cultured in the presence of hCG. Western blot analyses using antibodies against a protease domain revealed that the catalytically inactive 102-kDa proteins in the pars recta granules yield 66-kDa catalytically active and 82- and 59-kDa inactive molecules. We propose that the oviductin translated as 107.6-kDa precursors are processed both N- and C-terminally to give rise to a 66-kDa active form comprising a serine protease and two CUB domains.

Amino Acid Sequence↗

An efficient and cost effective nuclear medicine image network.

An image network that is in use in a large nuclear medicine department is described. This network was designed to efficiently handle a large volume of clinical data at reasonable cost. Small, limited function computers are attached to each scintillation camera for data acquisition. The images are transferred by cable network or floppy disc to a large, powerful central computer for processing and display. Cost is minimized by use of small acquisition computers not equipped with expensive video display systems or elaborate analysis software. Thus, financial expenditure can be concentrated in a powerful central computer providing a centralized data base, rapid processing, and an efficient environment for program development. Clinical work is greatly facilitated because the physicians can process and display all studies without leaving the main reading area.

Computer Communication Networks↗

Swedish nurses' attitudes towards research and development within nursing.

AIM OF THE STUDY: The aim was to develop and test an assessment instrument in order to study attitudes towards research and development within nursing among randomly selected professional nurses, registered nurses (RNs). BACKGROUND: The investigation was initiated because of the development within nursing and nursing education in Sweden towards better knowledge about research for the last four decades. METHODS: A questionnaire was designed, consisting of three parts: (1) demographic data, (2) attitude scale and (3) research awareness. Appropriate psychometric statistics were used such as factor analysis and parametric as well as nonparametric statistics to compare groups. The form was distributed to 407 randomly selected RNs. RESULTS: The response rate was 71% (n=289). The respondents were from four different examination years, representing four different nursing education systems in Sweden. Factor analysis (Maximum Likelihood and Oblimin rotation) gave seven factors with a total variance of 58% and Cronbach's alpha between 0.60 and 0.84. The factors were labelled 'Research language', 'Need of research knowledge', 'Participation', 'The Profession', 'Meaningfulness', 'Study literature' and 'Developing - Resources'. The results indicated that the respondents in general had positive attitudes towards nursing research. There were significant differences between the examined groups. The group examined in 1966 appeared as the least positive group. Forty-six per cent never or seldom utilized nursing research findings in their daily practice. About half of the respondents never read research reports. CONCLUSION: The results indicated that the new instrument is sensitive for measuring professional nurses' attitudes towards research and development in nursing. Even if the respondents had a positive attitude towards research and development, there was a poor application in their daily work. Further, the respondent's age, the year of RN examination and acquisition of research skills seemed to be of importance for the attitudes.

Adult↗

Preparation and characterization of Neisseria meningitidis mutants deficient in production of the human lactoferrin-binding proteins LbpA and LbpB.

Pathogenic members of the family Neisseriaceae produce specific receptors facilitating iron acquisition from transferrin (Tf) and lactoferrin (Lf) of their mammalian host. Tf receptors are composed of two outer membrane proteins, Tf-binding proteins A and B (TbpA and TbpB; formerly designated Tbp1 and Tbp2, respectively). Although only a single Lf-binding protein, LbpA (formerly designated Lbp1), had previously been recognized, we recently identified additional bacterial Lf-binding proteins in the human pathogens Neisseria meningitidis and Moraxella catarrhalis and the bovine pathogen Moraxella bovis by a modified affinity isolation technique (R. A. Bonnah, R.-H. Yu, and A. B. Schryvers, Microb. Pathog. 19:285-297, 1995). In this report, we characterize an open reading frame (ORF) located immediately upstream of the N. meningitidis B16B6 lbpA gene. Amino acid sequence comparisons of various TbpBs with the product of the translated DNA sequence from the upstream ORF suggests that the region encodes the Lf-binding protein B homolog (LbpB). The LbpB from strain B16B6 has two large stretches of negatively charged amino acids that are not present in the various transferrin receptor homologs (TbpBs). Expression of the recombinant LbpB protein as a fusion with maltose binding protein demonstrated functional Lf-binding activity. Studies with N. meningitidis isogenic mutants in which the lbpA gene and the ORF immediately upstream of lbpA (putative lbpB gene) were insertionally inactivated demonstrated that LbpA, but not LbpB, is essential for iron acquisition from Lf in vitro.

Amino Acid Sequence↗

Rapid acquisition of entire DNA polymerase gene of a novel herpesvirus from green turtle fibropapilloma by a genomic walking technique.

A 4837-bp sequence of a newfound green turtle herpesvirus (GTHV), implicated in the etiology of green turtle fibropapilloma, was obtained from tumor tissues of a green turtle with fibropapilloma using a genomic walking method based on restriction enzyme digestion, self-ligation and inverse polymerase chain reaction (IPCR). The 4837-bp sequence was 56.23% G/C rich and contained three nonoverlapping open reading frames (ORF). The largest ORF (3507-bp) encoded the DNA polymerase gene (pol gene), which exhibited a high degree of homology at both amino acid and nucleotide levels with the DNA pol genes of human and animal herpesviruses, with a predicted protein of 1169 amino acids and molecular weight of 132.6 kilodaltons. The ATG at 518 to 520 was the first initiation codon in the ORF and was presumed to be the first methionine codon of the pol gene. Phylogenetic analysis, based on the amino acid sequence of the GTHV DNA pol gene and the corresponding regions of other known human and animal herpesviruses, indicated that GTHV belonged to the Alphaherpesvirinae subfamily. The upstream ORF of the pol gene encoded the N-terminal region of the GTHV homologue of the DNA-binding protein gene, whereas the downstream ORF was the C-terminal region of a gene which was homologous to ORFs conserved in human and animal herpesviruses, i.e., herpes simplex virus 1 (HSV1) gene UL31, Epstein-Barr virus (EBV) gene BFLF2, equine herpesvirus 1 (EHV1) gene 29, and alcelaphine herpesvirus 1 (AHV1) hypothetical protein 69 gene. The arrangement of these three genes in GTHV genome was identical to that seen in other alphaherpesviruses. The sequence and location of the DNA pol gene in the GTHV genome should greatly facilitate future studies of the viral life cycle.

Alphaherpesvirinae↗