The Neonatal Pain, Agitation and Sedation Scale and the bedside nurse's assessment of neonates.

OBJECTIVE: To determine the reliability of an objective measure of pain, agitation and sedation using the Neonatal Pain, Agitation and Sedation Scale (N-PASS) compared with nursing bedside assessment. STUDY DESIGN: Neonates admitted in neonatal intensive care unit over a 6-month period were eligible. Pain and sedation were assessed with N-PASS, and a subjective questionnaire was administered to the bedside nurse. RESULT: A total of 218 neonates were eligible (median: gestational age 34.6 weeks, age at assessment 7 days). N-PASS pain score correlated significantly with both nurses' pain score (Spearman coefficient (r)=0.37; P<0.001) and agitation score (r=0.56; P<0.001). N-PASS sedation score correlated with nurses' sedation score (r=-0.39; P<0.001). Adjusting for gestational age, day of life, intrauterine drug exposure and use of high frequency ventilation only slightly attenuated the correlations (r=0.36, 0.55 and -0.31, respectively). CONCLUSION: The N-PASS captures nursing assessment of pain, agitation and sedation in this broad population and provides a quantitative assessment of subjective descriptions that often drives patient therapy.

First Report of Blueberry scorch virus on Highbush Blueberry in Connecticut and Massachusetts.

During June and July 2001, the Marucci Center received 33 foliage samples from healthy- and unhealthy-looking highbush blueberry (Vaccinium corymbosum L.) bushes from growers in Connecticut and Massachusetts, via local extension agents. Unhealthy bushes were reported to exhibit symptoms including leaf chlorosis and necrosis, blossom blight, tip dieback, or a general decline in vigor. Marginal leaf chlorosis, reddening, or necrosis characterized foliage samples from these bushes. Five-leaf samples from each bush were tested for Blueberry scorch virus (BlScV) (3) with Agri-Check detection kits (Hydros, Inc., Falmouth, MA). These kits use an indirect enzyme-linked immunosorbent assay (ELISA) protocol and antibodies developed at Rutgers University that are specific to the two eastern strains of BlScV (NJ1 and NJ2) (1). The ELISA extraction buffer was based on that used by Martin and Bristow (3) with 1% (wt/vol) nonfat dry milk powder added as a blocking agent. Fourteen samples from cvs. Blueray and Berkeley in both states and cvs. Elliott, Bluecrop, and Coville in Massachusetts tested positive for BlScV. These results were confirmed by a second test. Six of seven samples from symptomatic bushes and 8 of 26 samples from asymptomatic bushes harbored BlScV. Virus preparations extracted from five infected plants (two from Connecticut and three from Massachusetts) were examined using reverse transcription-polymerase chain reaction (RT-PCR) with oligonucleotide primers (5'-TGTGTCAAACAATATGGC-3' and 5'-GCATTTCGATGA-TTGCGG-3') designed to amplify a portion of the coat protein gene from any of the three known virus strains (1,2). Sequence analysis of fragments amplified from their coat protein genes revealed that two of the isolates from Massachusetts (GenBank Accession Nos. AY530957 and AY530958) and the two isolates from Connecticut (GenBank Accession Nos. AY530955 and AY530956) were similar but not identical to one another, and these four were most similar to strain NJ2. One isolate from Massachusetts (GenBank Accession No. AY530958) was most similar to strain NJ1. To our knowledge, this is the first report of BlScV on the east coast outside of New Jersey, where it was first reported in 1983 (4). These results indicate that the disease is now present in other blueberry-growing areas in the northeast and is likely to be spreading locally within those areas. Because blueberry scorch is symptomless in propagation material and may take several years to express symptoms in the field, the initial spread of the disease was probably due to the shipping of latently infected plants to BlScV-free areas before reliable testing was available. References: (1) T. D. Cavileer et al. J. Gen. Virol. 75:711, 1994. (2) B. T. Halpern and B. I. Hillman. Plant Dis. 80:219, 1996. (3) R. R. Martin and P. R. Bristow. Phytopathology 78:1636, 1988. (4) A. W. Stretch. (Abstr.) Phytopathology 73:375, 1983.

Evidence for interspecies transmission of viruses in natural populations of filamentous fungi in the genus Cryphonectria.

Interspecies transmission is a significant evolutionary event that has allowed a variety of pathogens to invade new host species. We investigated interspecies transmission of viruses between the chestnut blight fungus, Cryphonectria parasitica, and a sympatric unidentified Cryphonectria species in Japan. Two isolates of Cryphonectria sp. were found to contain Cryphonectria hypovirus 1 (CHV-1), which has been typically found in C. parasitica. Three lines of evidence support the hypothesis of interspecies transmission of CHV-1. First, host species occur sympatrically and therefore have the opportunity to come into physical contact. Second, we transmitted CHV-1 between species experimentally in the laboratory. Third, phylogenetic analysis of 476 bp of the ORF B region of CHV-1 showed that sequences from Cryphonectria sp. were more closely related to those from C. parasitica than to each other. Local geographical subdivision of virus sequences from both host species argues against the alternative hypothesis of independent evolution of CHV-1 since speciation of their hosts. Based on these findings, we rule out the hypotheses that CHV-1 diverged from viruses in a common ancestor of the hosts, or that ancestral polymorphisms in CHV-1 persisted in the two host taxa. Estimating the direction and frequency of interspecies transmission in nature will require more extensive samples of CHV-1 from both host species.

Cloning, sequencing, and promoter identification of Blueberry red ringspot virus, a member of the family Caulimoviridae with similarities to the "Soybean chlorotic mottle-like" genus.

The double-stranded DNA genome of Blueberry red ringspot virus (BRRV), a member of the family Caulimoviridae, was cloned and sequenced. The genome organization and relationships of the 8303 nt sequence revealed BRRV to be a tentative member of the genus that has been provisionally named "Soybean chlorotic mottle-like viruses", rather than a member of the genus Caulimovirus, in which it had been placed previously. Insertion of the putative 35S promoter homolog of BRRV into promoterless constructs carrying the UidA (beta-glucuronidase) gene resulted in high-level transient expression from cranberry and stable expression from transgenic tobacco. Sequences of 5'-RACE clones derived from transcripts from transgenic tobacco were consistent with the map position of the promoter.

Crypt1, an active Ac-like transposon from the chestnut blight fungus, Cryphonectria parasitica.

A moderately repetitive element was identified previously in the nuclear genome of the chestnut blight fungus, Cryphonectria parasitica, and has been used as a probe for population studies of the fungus. We report here that the repetitive element is a class II transposon of the hAT family of Activator (Ac)-like transposable elements. The element, named Crypt1, has a size of 3563 bp, including 21-bp terminal inverted repeats. A unique 8-bp direct repeat sequence flanking Crypt1 was identified in each of three clones examined. A single large ORF with the potential to encode a putative transposase of 946 amino acid residues was deduced from the sequence of Crypt1. Based on amino acid sequence alignments, Crypt1 is most closely related to other Ac-like transposons of filamentous ascomycetes. A single transcript of approximately 3.0 kb was identified by Northern hybridization experiments from Crypt1-containing isolates, suggesting that Crypt1 is an active element. An isolate containing a single, possibly defective, copy of Crypt1 was identified in C. parasitica isolates from China; no Crypt1 transcript was identified in this isolate. Transposition of Crypt1 was inferred from Southern and inverse PCR analyses of C. parasitica isolates maintained in the laboratory, but transposition appears to be a rare event.

Coding and reimbursement issues for the radiologist.

Radiologists are dependent on the proper execution of a complex administrative chain of disease and diagnosis coding and procedural coding to be properly reimbursed for the examinations they perform. The radiologist bears the ultimate responsibility for the appropriateness of these codes. However, many practicing radiologists are unaware of the critical link between the two coding systems and the systems that have developed to provide a common method of describing diseases, diagnoses, and procedures. This article is an introduction to these systems, and it emphasizes the importance of the involvement of the radiologist in coding. The inherent complexity of the reimbursement system in use is emphasized, as well as the essential role of the radiologist in complying with these often complicated and ever-changing directives.

Patients' role in the use of radiology testing for common office practice complaints.

BACKGROUND: Radiological studies are an important component of ambulatory medical costs, and guidelines often focus on their appropriate use. However, little is known about the correlates of the use of those services, particularly the influence of patients' preferences on physicians' utilization decisions. OBJECTIVES: To study patients presenting for outpatient treatment of respiratory problems and low back pain, and to examine the magnitude of the effect of the patients' perceived need for radiological studies (radiology preference score) on use of those services. DESIGN: Cross-sectional survey. SETTING: Office practices of generalist physicians in predominantly rural areas of 8 states. PARTICIPANTS: A total of 52 generalist physicians agreed to enroll consecutive Medicare-eligible patients making office visits for respiratory problems or low back pain. Of 1785 eligible patients invited to participate, 132 (7%) refused and 1137 (69%) of 1653 returned questionnaires. MEASUREMENTS: Radiology utilization rates (plain film, computed tomographic scan, or magnetic resonance image scan) were determined by patient self-report. To assess perceived need for radiological studies, we asked patients how necessary they believed an x-ray film was in the evaluation of 4 common complaints (respiratory problems, low back pain, knee pain, and knee swelling). A summary radiology preference score was created from 3 of the 4 items, excluding the item referring to the patients' index complaint. RESULTS: Six hundred fifteen respiratory and 522 low back pain patients were enrolled; mean ages were 69 and 64 years, respectively. Radiology utilization rates were 37% for respiratory and 26% for low back pain patients. In multiple logistic regression models, for respiratory patients radiology utilization was related significantly to the radiology preference score (odds ratio [OR] for fourth quartile compared with first quartile, 1.94; 95% confidence interval [CI], 1.11-3.37; P = .02), to having a physician who owned radiology equipment (OR, 1.81; 95% CI, 1.23-2.66; P = .002), and current smoking (OR, 1.58; 95% CI, 1.04-2.41; P = .03). For low back pain patients, radiology utilization was significantly related to the radiology preference score (OR for fourth compared with first quartile, 2.55; 95% CI, 1.29-5.06; P = .007), bothersomeness of the pain (OR for fourth compared with first quartile, 3.74; 95% CI, 1.74-8.04; P<.001), and a diagnosis of osteoporosis (OR, 1.67; 95% CI, 1.01-2.75; P = .04). CONCLUSIONS: Patients' perceived need for radiological studies was significantly associated with use of those services for outpatients with respiratory problems and low back pain. These findings suggest that patients communicate their wishes to physicians, either directly or indirectly, regarding services they think are necessary. Differences in physicians' adherence to guidelines regarding radiology utilization may in part reflect variations in patients' perceived need for those services. Efforts to educate patients about when radiological studies are medically indicated may be an important complement to practice guidelines or other utilization-related financial incentives.

In vitro translational analysis of genomic, defective, and satellite RNAs of Cryphonectria hypovirus 3-GH2.

Cryphonectria hypovirus 3-GH2 (CHV3-GH2) is a member of the fungal virus family Hypoviridae that differs from previously characterized members in having a single large open reading frame with the potential to encode a protein of 326 kDa from its 9.8-kb genome. The N-terminal portion of the ORF contains sequence motifs that are somewhat similar to papain-like proteinases identified in other hypoviruses. Translation of the ORF is predicted to release autocatalytically a 32.5-kDa protein. A defective RNA, predicted to encode a 91.6-kDa protein representing most of the N-terminal proteinase fused to the entire putative helicase domain, and two satellite RNAs, predicted to encode very small proteins, also are associated with CHV3-GH2 infected fungal cultures. We performed in vitro translation experiments to examine expression of these RNAs. Translation of three RT-PCR clones representing different lengths of the amino-terminal portion of the ORF of the genomic RNA resulted in autocatalytic release of the predicted 32.5-kDa protein. Site-directed mutagenesis was used to map the processing site between Gly(297) and Thr(298). In vitro translation of multiple independent cDNA clones of CHV3-GH2-defective RNA 2 resulted in protein products of approximately 92 kDa, predicted to be the full-length translation product, 32 kDa, predicted to represent the N-terminal proteinase, and 60 kDa, predicted to represent the C-terminal two-thirds of the full-length product. In vitro translation of cDNA clones representing satellite RNA 4 resulted in products of slightly less than 10 kDa, consistent with the predicted 9.4 kDa product.

Satellite and defective RNAs of Cryphonectria hypovirus 3-grand haven 2, a virus species in the family Hypoviridae with a single open reading frame.

Cryphonectria parasitica hypovirus 3-Grand Haven 2 (CHV3-GH2) is the most recently characterized member of the Hypoviridae family of viruses associated with hypovirulence of the chestnut blight fungus. Isolates of CHV3-GH2 contain either three or four double-stranded (ds) RNAs that are visible on ethidium bromide-stained agarose or polyacrylamide gels. Only the largest dsRNA appears to be required for virus infectivity, and was characterized previously (C. D. Smart et al., 1999, Virology 265, 66-73). In this study, we report the cloning, sequencing, and analysis of the other three dsRNAs. Sizes of the accessory dsRNAs are 3.6 kb (dsRNA 2), 1.9 kb (dsRNA 3), and 0.9 kb (dsRNA 4), compared to 9.8 kb for the genomic dsRNA segment (dsRNA 1). All three accessory dsRNA species are polyadenylated on the 3'-end of one strand, as is genomic dsRNA. DsRNA 2 represents a defective form of dsRNA 1, with the 5'-terminal 1.4 kb derived from the 5'-end of dsRNA 1 and the 3'-terminal 2.2 kb from the 3'-end of dsRNA 1. A single major open reading frame (ORF) is evident from deduced translations of dsRNA 2. The deduced translation product is a 91-kDa protein that represents a fusion consisting of the entire N-terminal protease and the entire putative helicase domain. DsRNAs 3 and 4 represent satellite RNAs that share very little sequence with dsRNA 1 and 2. DsRNA 4 is 937 nucleotides, excluding the poly(A)(+). The first AUG of the polyadenylated strand of dsRNA 4 occurs eight residues in from the 5'-terminus and would initiate the largest ORF on dsRNA 4, with the coding capacity for a 9.4-kDa protein. Within the deduced ORF and approximately 100 nucleotides from the 5'-end of dsRNA 4 is a 22-base sequence that is identical to sequences found in the nontranslated leaders of dsRNAs 1 and 2. DsRNA 3 accumulation in infected cultures varied, but it was less abundant than dsRNA 4. DsRNA 3 was found to represent a head-to-tail dimer of dsRNA 4 linked by a poly(A)/(U) stretch of 40-70 residues.

Elevation of nitrotyrosine and nitrate concentrations in cystic fibrosis sputum.

Nitric oxide (NO) is increased in the exhaled air of some patients with inflammatory lung disorders, but not in others. NO may combine with superoxide to form peroxynitrite, which lowers NO gas concentrations, increases formation of nitrate, and increases nitration of tyrosine residues on proteins. We hypothesized that superoxide released from neutrophils in the lower respiratory tract of cystic fibrosis (CF) results in increased nitrate and nitrotyrosine levels in sputum. In order to test this hypothesis, exhaled NO was collected from 5 stable adult CF subjects and from 5 nonsmoking normal controls. Consistent with previous reports, exhaled NO concentrations were not increased in CF exhaled air (22.6 +/- 1.5 ppb vs. 28.6 +/- 1.5 ppb in normals, P > 0.05). Sputum was collected from 9 adult CF subjects and the same 5 normal controls and evaluated for nitrite, nitrate, and nitrotyrosine. Nitrate and nitrotyrosine levels, but not nitrite, were significantly elevated in CF. Recently, myeloperoxidase has also been implicated as a mechanism of nitrotyrosine formation. Therefore, myeloperoxidase was measured and found to be elevated in the CF sputum (64.2 +/- 35.9 vs. 0.73 +/- 0.16 U/mL, P < 0.001), and was found to correlate with concentrations of nitrotyrosine (r = 0.87, P < 0.05). However, in vitro studies with myeloperoxidase and murine lung epithelial cells did not demonstrate a reduction of NO gas with nitrotyrosine or an increase in nitrate formation. These data demonstrate that nitrate and nitrotyrosine are elevated in the sputa of CF subjects and suggest increased production of NO in the lower respiratory tract of CF patients, despite the relatively low exhaled NO levels. Pediatr Pulmonol. 2000; 30:79-85. Published 2000 Wiley-Liss, Inc.

Evaluation of a mandatory radiology resident research rotation.

RATIONALE AND OBJECTIVES: This purpose of this study was to determine if a 1-month, mandatory research rotation results in resident interest and productivity in research. MATERIALS AND METHODS: A survey composed primarily of closed-end questions was constructed to evaluate the opinions and experience of 24 residents who completed the research rotation during a 3 1/2-year period. RESULTS: All 24 residents filled out the survey. Seventeen residents did clinical projects, two did bench research, and two did projects that were socioeconomic or legal in nature. Thirteen residents presented data at a meeting, and three of these presentations won awards. Resident research projects resulted in 18 manuscripts at the time of the survey (seven published, eight in preparation, and three in revision or review). Seventeen residents thought the experience was worthwhile and the research month should be continued; two residents did not. Ten residents reported that in retrospect, they would have done the rotation even if it had not been required, and eight reported the experience increased their appreciation of the value of research in medicine. CONCLUSION: A 1-month, mandatory resident research rotation produces resident interest in research and enhances departmental research productivity.

The past 25 years in medical imaging research: a memoir.

During the past 25 years, medical imaging research has progressed in both scope and quality. Factors intrinsic to the specialty and changes occurring in medicine and society have fostered imaging research development. The advent of new, computer-based technologies that can be brought to bear on research, the increasing sophistication of researchers, and the greater availability of extramural funding have been primary factors in the promulgation of research improvements. Radiology researchers have the opportunity to play an important role in the genesis of the molecular medicine of the future. Whether they do so is dependent on whether radiologists identify necessary resources, new researchers receive appropriate training, and investigators are willing to think differently than they have in the past about the capabilities of imaging.

Cryphonectria hypovirus 3, a virus species in the family hypoviridae with a single open reading frame.

Isolate Grand Haven (GH) 2 is a naturally occurring isolate of the chestnut blight fungus, Cryphonectria parasitica, that is greatly reduced in virulence due to the presence of a double-stranded RNA virus. Unlike many other virus-infected, hypovirulent isolates, GH2 is not substantially reduced in pigmentation, conidiation, or laccase expression compared to its virus-free counterpart. The dsRNA genome of the GH2 virus was cloned, sequenced, and compared to hypovirulence-associated viruses of the family Hypoviridae. GH2 dsRNA is considerably smaller than previously characterized members of the family, 9.8 kb compared to 12.5-12.7 kb for other members. The genome organization of GH2 dsRNA reflected the substantial difference in genome size. Like other members of the family, one strand contained a poly(A)(+) tail at the 3' end and a long sequence with several minicistrons at the 5' end of the same strand. Only a single open reading frame (ORF) of 8622 nucleotides was predicted from deduced translations of the poly(A)(+)-containing strand, however. This contrasts with the two-ORF structures of previously characterized members. Analysis of the deduced ORF of GH2 dsRNA revealed putative proteinase, RNA polymerase, and helicase domains similar to those previously identified in confirmed members of the virus family Hypoviridae. GH2 dsRNA was more distantly related to Cryphonectria hypovirus (CHV) 1-EP713 and CHV2-NB58 than the latter two were to each other but has features in common with each of those viruses. We propose that the GH2 virus be included in this taxon as a member of the genus Hypovirus, representing a strain of a new species, CHV3.

Fungal proteinase expression in the interaction of the plant pathogen Magnaporthe poae with its host.

Infection by pathogenic fungi involves breaching the outer layer of the host by either mechanical or enzymatic means. Subtilisin-like proteinases are considered to be important in the infection process of entomopathogenic, nematophagous, and mycoparasitic fungi. Little is known regarding the expression of such proteinases by plant pathogenic fungi. Magnaporthe poae, a fungal pathogen of Kentucky bluegrass, expressed a subtilisin-like proteinase, proteinase Mp1, in the infected roots. Antibody was produced against the purified enzyme. From immunoblot analysis, expression of the proteinase in infected roots correlated with increasing severity of disease symptoms. Sequence analysis of a genomic clone indicated proteinase Mp1 was homologous to other fungal subtilisin-like proteinases. DNA gel blot analysis indicated proteinase Mp1 was encoded by a small gene family.

The RSNA-AUR-ARRS introduction to research program for 2nd year radiology residents: effect on career choice and early academic performance. Radiological Society of North America. Association of University Radiologists. American Roentgen Ray Society.

PURPOSE: To evaluate graduates of the first 5 years of the Introduction to Research program, sponsored by the Radiological Society of North America, Association of University Radiologists, and American Roentgen Ray Society, to determine if they more frequently entered and demonstrated early success in academic careers than did nonparticipants. MATERIALS AND METHODS: In 1997, the authors surveyed all 396 program participants for the 1990-1995 period and 2,047 nonparticipants who were in the same residencies as the participants during that period. Career plans, research participation, and academic achievements were compared. RESULTS: The survey was completed by 321 participants (81%) and 1,153 nonparticipants (56%). More than a third of participants thought the program had a strong effect on their initial career decision. Forty-one percent of participants who completed training (n = 192) were currently in academia versus 26% of nonparticipants (n = 676; P = .001). Of the 321 participants, 35% believed they would spend most of their careers in academia and 15% thought research would be their major focus versus 19% and 8% for the 1,153 nonparticipants (P < .001). Participants had higher levels of academic achievement earlier in their careers than did nonparticipants. CONCLUSION: The Introduction to Research program is encouraging the development of successful research careers. However, the results may be influenced by the self- or departmental selection of participants who were oriented toward research independent of program participation.