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Intuitively, vagueness involves some sort of indeterminacy: if Plato is a borderline case of baldness, then there is no fact of the matter about whether or not he’s bald—he’s neither bald nor not bald. The leading formal treatments of such indeterminacy—three valued logic, supervaluationism, etc.—either fail to validate the classical theorems, or require that various classically valid inference rules be restricted. Here we show how a fully classical, yet indeterminist account of vagueness can be given within natural semantics, an alternative semantics for classical proof theory. The key features of the account are: there is a single notion of truth—definite truth—and a single notion of validity; sentences can be true, false, or undetermined; all classical theorems and all classical inference rule are valid; the sorites argument is unsound; ‘definitely’ is treated as a meta-language predicate; higher-order vagueness is handled via semantic ascent.

Objectives: To evaluate immediate placement and immediate restoration of a novel implant with a 12°-angled prosthodontic platform, in fresh extraction sockets of the aesthetic zone of the maxilla. Materials and methods: Tapered, roughened surface implants of 4 mm (n=15) or 5 mm (n=13) diameter were placed in 27 participants (mean age: 47.1 years; range: 21–71 years) requiring an immediate replacement of single anterior maxillary teeth. Provisional screw-retained all-ceramic crowns were placed within 4 h following optical impressions. At 8 weeks (baseline), definitive screw-retained all-ceramic crowns were placed in occlusion. Results: Twenty-six of the 28 implants met the inclusion criteria at surgery. Marginal bone levels revealed bone gain between surgery and baseline, and between baseline and 1 year of 0.2 mm (SD 0.75) and 0.78 mm (SD 2.45). Mean mid-buccal mucosal margins showed gains of 0.2 mm (SD 0.44). Prosthodontic maintenance and the aesthetics of the screw-retained implant crowns were facilitated by the external hex 12°-angled prosthodontic platform on the novel implant design, re-orientating the access cavity to the palatal or occlusal surfaces. All-ceramic implant crowns showed a high success rate with low maintenance issues over 1 year. Conclusion: Tapered, roughened-surfaced implants with a novel 12°-angled prosthodontic platform immediately placed in fresh extraction sockets, immediately restored with provisional crowns and subsequent definitive crowns at 8 weeks were successful for 1 year. To cite this article: Brown SDK, Payne AGT. Immediately restored single implants in the aesthetic zone of the maxilla using a novel design: 1-year report. Clin. Oral Impl. Res. 22, 2011; 445–454.

The interaction between echovirus 11 strain 207 (EV11-207) and decay-accelerating factor (DAF or CD55) at the apical surface of polarized Caco-2 cells results in rapid transport of the virus to tight junctions and in its subsequent uptake. A virus mutant (EV11-207R) which differs at 6 amino acids and whose affinity for DAF is apparently significantly lower remains at the apical surface, from where its uptake occurs. Binding of EV11-207 to DAF and its transport to tight junctions result in a loss of function of the junctions. In contrast, the mutant virus EV11-207R is not transferred to tight junctions, nor does it impair the integrity of these junctions. Cholesterol depletion from the apical membrane leads to DAF aggregation and, presumably, internalization and inhibits infection by EV11-207. However, infection by EV11-207R is significantly less sensitive to cholesterol depletion than infection by EV11-207, confirming the DAF requirement for EV11-207, but not EV11-207R, to infect cells. These data strongly indicate that in the case of infection of polarized epithelial cells by echovirus 11, DAF binding appears be a key determinant in the choice of entry pathway, at least in cell culture.

Coupled expression of the M1 and BM2 open-reading frames (ORFs) of influenza B from the dicistronic segment 7 mRNA occurs by a process of termination-dependent reinitiation. The AUG start codon of the BM2 ORF overlaps the stop codon of the upstream M1 ORF in the pentanucleotide UAAUG, and BM2 synthesis is dependent upon translation of the M1 ORF and termination at the stop codon. Here, we have investigated the mRNA sequence requirements for BM2 expression. Termination–reinitiation is dependent upon 45 nucleotide (nt) of RNA immediately upstream of the UAAUG pentanucleotide, which includes an essential stretch complementary to 18S rRNA helix 26. Thus, similar to the caliciviruses, base-pairing between mRNA and rRNA is likely to play a role in tethering the 40S subunit to the mRNA following termination at the M1 stop codon. Consistent with this, repositioning of the M1 stop codon more than 24 nt downstream from the BM2 start codon inhibited BM2 expression. RNA structure probing revealed that the RNA upstream of the UAAUG overlap is not highly structured, but upon encountering the M1 stop codon by the ribosome, a stem–loop may form immediately 5′ of the ribosome, with the 18S rRNA complementary region in the apical loop and in close proximity to helix 26. Mutational analysis reveals that the normal requirements for start site selection in BM2 expression are suspended, with little effect of initiation codon context and efficient use of noncanonical initiation codons. This suggests that the full complement of initiation factors is not required for the reinitiation process.

We present a simple numerical optimization procedure to search for highly entangled states of 2, 3, 4 and 5 qubits. We develop a computationally tractable entanglement measure based on the negative partial transpose criterion, which can be applied to quantum systems of an arbitrary number of qubits. The search algorithm attempts to optimize this entanglement cost function to find the maximal entanglement in a quantum system. We present highly entangled 4-qubit and 5-qubit states discovered by this search. We show that the 4-qubit state is not quite as entangled, according to two separate measures, as the conjectured maximally entangled Higuchi–Sudbery state. Using this measure, these states are more highly entangled than the 4-qubit and 5-qubit GHZ states. We also present a conjecture about the NPT measure, inspired by some of our numerical results, that the single-qubit reduced states of maximally entangled states are all totally mixed.

We have used X-ray crystallography to determine the structure of a decay accelerating factor (DAF)-binding, clinic-derived isolate of echovirus 11 (EV11-207). The structures of the capsid proteins closely resemble those of capsid proteins of other picornaviruses. The structure allows us to interpret a series of amino acid changes produced by passaging EV11-207 in different cell lines as highlighting the locations of multiple receptor-binding sites on the virion surface. We suggest that a DAF-binding site is located at the fivefold axes of the virion, while the binding site for a distinct but as yet unidentified receptor is located within the canyon surrounding the virion fivefold axes.

The Beaudette strain of IBV was passaged 16 times in chick kidney cells. Total cellular RNA was analyzed by Northern hybridization and was probed with 32 P-labeled cDNA probes corresponding to the first 2 kb of the 5′ end of the genome, but excluding the leader, and to the last 1.8 kb of the 3′ end of the genome. A new, defective IBV RNA species (CD-91) was detected at passage 6. The defective RNA, present in total cell extract RNA and in oligo-(dT) 30 -selected RNA from passage 15, was amplified by the reverse transcription-polymerase chain reaction (RT-PCR) to give four fragments. The oligonucleotides used were selected such that CD-91 RNA, but not the genomic RNA, would be amplified. Cloning and sequencing of the PCR products showed that CD-91 comprises 9.1 kb and has three regions of the genome. It contains 1133 nucleotides from the 5′ end of the genome, 6322 from gene 1b corresponding to position 12,423 to 18,744 in the IBV genome, and 1626 from the 3′ end of the genome. At position 749 one nucleotide, an adenine residue, was absent from CD-91 RNA. By Northern hybridization CD-91 RNA was detected in virions in higher amounts than the subgenomic mRNAs.

The Marine Forensics Panel of the Society of Naval Architects and Marine Engineers (SNAME) grew out of analyses of the shipwrecks of the German battleshipBismarck and the passenger shipsTitanic andLusitania. It is now co-sponsored by five other professional societies on both sides of the Atlantic. A number of ship losses have been investigated, and lessons have been learned relevant both to marine technology and history.

Previous studies in vitro of the processing of cloned polyprotein fragments from the coronavirus infectious bronchitis virus (IBV) large open reading frame (ORF1), confirmed the activity of a predicted 3C-like proteinase (3CLP) domain and suggested that the proteinase is released autocatalytically from the polyprotein in the form of a 35 kDa protein, 3CLpro, capable of further cleavages in trans. In order to identify such cleavages within the ORF1 polyprotein mediated by 3CLpro, the proteinase was expressed in bacteria, purified and used in trans cleavage assays with polyprotein fragments lacking the 3CLP domain as targets. The proteinase was expressed as a polyprotein fragment which was able to process during expression in bacterial cells, releasing mature 3CLpro. A histidine (His6) tag was introduced close to the C-terminus of the proteinase to aid purification. Processing demonstrated by the tagged proteinase was indistinguishable from that of the wild-type enzyme indicating that the site chosen for the tag was permissive. From these studies we were able to demonstrate trans cleavages consistent with the use of most of the previously predicted or identified sites within the open reading frame of gene 1. This tentatively completes the processing map for the ORF1 region with respect to 3CLpro.

Proteolytic processing of the polyprotein encoded by mRNA 1 is an essential step in coronavirus RNA replication and gene expression. We have previously reported that an open reading frame (ORF) 1a-specific proteinase of the picornavirus 3C proteinase group is involved in processing of the coronavirus infectious bronchitis virus (IBV) 1a/1b polyprotein, leading to the formation of a mature viral protein of 100 kDa. We report here the identification of a novel 10-kDa polypeptide and the involvement of the 3C-like proteinase in processing of the ORF 1a polyprotein to produce the 10-kDa protein species. By using a region-specific antiserum, V47, raised against a bacterial-viral fusion protein containing IBV sequence encoded between nucleotides 11488 and 12600, the 10-kDa polypeptide was detected in lysates from both IBV-infected and plasmid DNA-transfected Vero cells. Coexpression, deletion, and mutagenesis studies showed that this novel polypeptide was encoded by ORF 1a from nucleotide 11545 to 11878 and was cleaved from the 1a polyprotein by the 3C-like proteinase domain. Evidence presented suggested that a previously predicted Q-S (Q3783 S3784) dipeptide bond encoded by ORF 1a between nucleotides 11875 and 11880 was responsible for the release of the C terminus of the 10-kDa polypeptide and that a novel Q-N (Q3672 N3673) dipeptide bond encoded between nucleotides 11542 and 11547 was responsible for the release of the N terminus of the 10-kDa polypeptide.

The construction of a full-length clone of the avian coronavirus infectious bronchitis virus (IBV) defective RNA (D-RNA), CD-91 (9,080 nucleotides [Z. Penzes et al., Virology 203:286-293]), downstream of the bacteriophage T7 promoter is described. Electroporation of in vitro T7-transcribed CD-91 RNA into IBV helper virus-infected primary chick kidney cells resulted in the production of CD-91 RNA as a replicating D-RNA in subsequent passages. Three CD-91 deletion mutants were constructed--CD-44, CD-58, and CD-61--in which 4,639, 3,236, and 2,953 nucleotides, respectively, were removed from CD-91, resulting in the truncation of the CD-91 long open reading frame (ORF) from 6,465 to 1,311, 1,263, or 2,997 nucleotides in CD-44, CD-58, or CD-61, respectively. Electroporation of in vitro T7-transcribed RNA from the three constructs into IBV helper virus-infected cells resulted in the replication and packaging of CD-58 and CD-61 but not CD-44 RNA. The ORF of CD-61 was further truncated by the insertion of stop codons into the CD-61 sequence by PCR mutagenesis, resulting in constructs CD-61T11 (ORF: nucleotides 996 to 1,058, encoding 20 amino acids), CD-61T22 (ORF: nucleotides 996 to 2,294, encoding 432 amino acids), and CD-61T24 (ORF: nucleotides 996 to 2,450, encoding 484 amino acids), all of which were replicated and packaged to the same levels as observed for either CD-61 or CD-91. Analysis of the D-RNAs showed that the CD-91- or CD-61-specific long ORFs had not been restored. Our data indicate that IBV D-RNAs based on the natural D-RNA, CD-91, do not require a long ORF for efficient replication. In addition, a 1.4-kb sequence, corresponding to IBV sequence at the 5' end of the 1b gene, may be involved in the packaging of IBV D-RNAs or form part of a cis-acting replication element.

This paper represents the final forensic analyses concerning the losses of the passenger liners RMS Titanic and Lusitania based on photographic and other evidence taken from their wreck sites. It will detail the role of brittle fracture and notch sensitivity in the Titanic's steel plates that contributed to her loss on 14–15 April 1912 and the possibility of a similar occurrence in the Lusitania. The conclusions reached here are based upon six expeditions to the Titanic wreck site over a period of nine years, commencing with the Ballard-Michel Expedition in 1985 and ending with the 1994 IFREMER Expedition. The Lusitania analysis will reveal what brought about her loss on 7 May 1915, based upon the Ballard Expedition of 1993 and our own research into her loss, propulsion plant, and hull design.

We have reviewed the results of ^sup 99m^technetium-methylene diphosphonate (Tc-MDP) bone scintigrams performed on patients following total knee arthroplasty. In addition, p 67 gallium (Ga) citrate scintigrams were carried out sequentially on 29 patients. Three groups of patients were identified: those with asymptomatic knees (undergoing scans for other reasons); those with aseptic or septic loosening; and those with pain without radiologic evidence of loosening. There was good correlation between the results of the scans and the final outcome. We conclude that sequential ^sup 99m^Tc-MDP and p 67 Ga citrate scintigrams are useful for demonstrating the presence of aseptic and septic loosening in knee prostheses, and pain with a normal scan appearance is probably not due to loosening or infection.

Coronavirus gene expression involves proteolytic processing of the mRNA 1-encoded polyproteins by viral and cellular proteinases. Recently, we have demonstrated that an ORF 1b-encoded 100-kDa protein is proteolytically cleaved from the 1a/1b fusion polyprotein by a viral-specific proteinase of the picornavirus 3C proteinase group (3C-like proteinase). In this report, the 3C-like proteinase has been further analysed by internal deletion of a 2.3-kb fragment between the 3C-like proteinase-encoding region and ORF 1b and by substitution mutations of its catalytic centre as well as the two predicted cleavage sites flanking the 100-kDa protein. The results show that internal deletion of ORF 1a sequences from nucleotide 9911 to 12227 does not influence the catalytic activity of the proteinase in processing of the 1a/1b polyprotein to the 100-kDa protein species. Site-directed mutagenesis studies have confirmed that the predicted nucleophilic cysteine residue (Cys2922) and a histidine residue encoded by ORF 1a from nucleotide 8985 to 8987 (His2820) are essential for the catalytic activity of the proteinase, and that the QS(G) dipeptide bonds are its target cleavage sites. Substitution mutations of the third component of the putative catalytic triad, the glutamic acid 2843 (Glu2843) residue, however, do not affect the processing to the 100-kDa protein. In addition, cotransfection experiment shows that the 3C-like proteinase is capable of trans-cleavage of the 1a/1b polyprotein. These studies have confirmed the involvement of the 3C-like proteinase domain in processing of the 1a/1b polyprotein, the predicted catalytic centre of the proteinase, and its cleavage sites.

Nucleotide sequence analysis has shown previously that the genomic-length mRNA (mRNA1) of the coronavirus infectious bronchitis virus (IBV) contains two large open reading frames (ORFs), 1a and 1b, with the potential to encode polyproteins of approximately 441 and 300 kDa, respectively. We have characterized the specificity of a set of region-specific antisera raised against the 5′-portion of ORF 1a by immunoprecipitation of in vitro-synthesized, C-terminally truncated 1a polypeptides and used these antisera to detect virus-specific proteins in IBV-infected Vero cells. Two antisera, which had specificity for IBV sequences from nucleotides 710 to 2079 and 1355 to 2433, respectively, immunoprecipitated a polypeptide of approximately 87 kDa from IBV-infected Veto cells. In vitro translation of ORF 1a sequence terminating at nucleotide 5763 did not produce this protein unless the in vitro translation products were incubated with Vero cell S10 extracts prepared from either IBV-infected or mock-infected Vero cells. However, processing of the 87-kDa protein was also observed when the same region was expressed in Vero cells using the vaccinia virus/T7 expression system. This observation indicates that the 87-kDa polypeptide is encoded within the 5′-most 3000 nucleotides of mRNA 1 and that it might be cleaved from the 1a polyprotein by viral and cellular proteinases.

The genomic RNA of human astrovirus serotype 1 (HAst-1) contains three open reading frames (ORFs), 1a, 1b, and 2. ORF 1b is located downstream of, and overlaps, 1a, and it has been suggested on the basis of sequence analysis that expression of ORF 1b is mediated through -1 ribosomal frameshifting. To examine this possibility, a cDNA fragment containing the 1a-1b overlap region was cloned within a reporter gene and placed under the control of the bacteriophage SP6 promoter in a recombinant plasmid. Synthetic transcripts derived from this plasmid, when translated in the rabbit reticulocyte lysate cell-free system, specified the synthesis of polypeptides whose size and antibody reactivity were consistent with an efficient -1 ribosomal frameshift event at the overlap region. The HAst-1 frameshift signal has two essential components, a heptanucleotide slippery sequence, A6C, and a stem-loop structure in the RNA. The presence of this structure was confirmed by complementary and compensatory mutation analysis and by direct structure probing with single- and double-stranded RNA-specific reagents. The HAst-1 frameshift signal, like that present at the overlap of the gag and pro genes of the retrovirus human T-cell lymphotrophic virus type II, does not involve the formation of an RNA pseudoknot.

We have determined the complete genomic sequence of human astrovirus serotype 1 isolated in Newcastle upon Tyne. The genome is 6813 nucleotides long and contains three sequential open reading frames (ORFs). The two closest to the 5' end are linked by a ribosomal frameshifting motif and contain sequence motifs indicative of non-structural virus proteins: serine protease and RNA-dependent RNA polymerase. A nuclear addressing sequence is also located here. The 3' ORF encodes the virion structural polypeptides as a polyprotein precursor. This genomic organization resembles that of the plant virus family Luteoviridae.

A recombinant fowlpox virus was produced which expressed the fusion protein (F) of turkey rhinotracheitis virus (TRTV), a pneumovirus. Turkey poults were vaccinated twice, at an interval of 2 weeks, intramuscularly and by wing web on each occasion, with the recombinant or a control fowlpox virus. Two weeks after the second vaccination the poults were challenged superconjunctivally and intranasally with virulent TRTV. A partially protective immune response was achieved; turkeys vaccinated with the F recombinant showed milder clinical signs and 1000-fold less challenge virus was recovered from the nose and trachea compared with turkeys that had been vaccinated with control fowlpox virus. Expression of the F protein induced antibodies which were detectable both by an ELISA and a virus neutralization test. These results show that the immune responses to the F protein play a major role in protection against TRTV and indicate that recombinant viruses expressing the TRTV F protein have potential as vaccines against TRT.

The following Brief Report was written by residents. A discussion by a member of the residents' faculty follows. We invite any resident to submit such articles, together with commentary by a faculty member.

Negative-stranded virion RNA and oligonucleotide primers complementary to fusion (F) protein gene sequences were used to generate cDNA clones, revealing that the gene 5′-proximal to the F protein corresponded to the M2 (22K) gene, as in respiratory syncytial (RS) virus. The transcription start signal, GGGACAAGU, was identical to that of the F and matrix (M) proteins of turkey rhinotracheitis virus (TRTV). There were two sequences with the potential to function as transcription termination/poly(A) signals, located at nucleotides 751 to 762 and 777 to 787; 15 clones derived from mRNA indicated that the first of these sequences formed the major signal. Part of the next downstream (5′) gene was sequenced; unlike mammalian pneumoviruses the TRTV M2 gene did not overlap the beginning of the 5′-proximal gene. Northern blotting indicated that infected Vero cells contained less M2 mRNA than F mRNA and that about half of the M2 mRNA was present as a F-M2 dicistronic mRNA. The M2 gene contained two overlapping open reading frames (ORFs 1 and 2), as with RS virus. ORF 1 comprised 558 nucleotides with the coding potential for a 186 amino acid polypeptide, M r 20959, eight or nine residues shorter than for human RS virus strains. The overall amino acid identity was 40%, the N-terminal one-third of the proteins sharing 62% of residues, the remainder 29%. A hydropathy plot of the TRTV M2 protein had close similarity to that of the M2 of RS virus. The protein was predicted to have a basic character with no N-terminal signal sequence or other major highly hydrophobic sequences. In vitro translation of a transcript comprising both ORFs 1 and 2 produced a single product of apparent M r 23000, corresponding to the M2 product of ORF 1. Site-directed mutagenesis confirmed that this product was derived from ORF 1 and that frameshifting was not involved. The second ORF was expressed only from a transcript which lacked the AUG codons of ORF 1 and, although occupying a similar position to that in the RS virus M2 gene, had virtually no amino acid identity in its 73 residue length and was approximately 25% shorter than the corresponding RS virus ORF 2. The hydropathy plot of the potential products of the second ORFs of TRTV and RS virus showed little resemblance. Taken together these results suggest that ORF 2 is unlikely to be expressed in vivo. Our accumulated data show that TRTV has the partial gene order 3′ M-F-M2 5′, whereas the corresponding RS virus genes are arranged 3′ M-SH-G-F-M2 5′.

Describes the structural failures in the RMS Titanic and the ro/spl circ/le of brittle fracture in her steel plates that contributed to her loss on 14-15 April 1912. The conclusions reached are based upon five expeditions to the wreck site over a period of eight years, commencing with the Ballard-Michel expedition in 1985. >

Distortion product otoacoustic emission (DPOAE) measurements are becoming popular in the clinical realm because they have been shown to reflect cochlear function. The primary tones used to evoke the DPOAE are important in determining the amplitude of the emission recorded in the ear canal. This study examined the ratio of the primaries necessary to determine the maximum amplitude emission as a function of development, stimulus level and frequency. Optimum f(2)/f(1) ratios were measured utilizing the f(1)-sweep technique from 105 neonates between 30-42 weeks conceptional age (CA) and 40 adults. No significant difference for optimum ratio was shown between the neonatal and the adult groups. Primary tone frequency had a significant effect on optimum ratio for both neonates and adults. Low f(2) frequencies (4 kHz) were associated with higher optimum ratios than high f(2) frequencies (4 kHz). The adult group was used to investigate the effect of stimulus level on the optimum f(2)/f(1) ratio for f(2) frequencies from 1.7 to 10 kHz. Regression analysis showed significant differences across levels of the primaries at all frequencies except for f(2)=3.4 and 7.0 kHz. These differences in f(2)/f(1) ratio across stimulus frequency and level may be attributed to the change in the shape of the excitation profiles along the basilar membrane.

A group of 48 patients, consecutive apart from exclusions, from a 1-year series of 60 cases of aneurysmal subarachnoid haemorrhage (SAH), was reviewed, in respect of clinical and radiological features, surgical management, clinical outcome, psychological distress and psychometric status, the neuropsychological assessment being compared with a closely-matched group of controls, the postoperative assessment being accompanied by a single positron emission computed tomogram (SPECT) scan. A review of those features which might have been expected to have a bearing on cognitive outcome (CT abnormalities at the outset, angiographic vasospasm, operative aneurysmal leakage, temporary vessel occlusion) failed to show a significant difference on cognitive tests. There was, however, a cognitive deficit shown by the patient group as a whole, when compared with the controls. Thus, SAH itself, the initial insult, would appear to be the essential factor in the production of persistent cognitive deficits.

We present here evidence demonstrating that four previously predicted Q-S(G) cleavage sites, encoded by the IBV sequences from nucleotide 15,129 to 15,134, 16,929 to 16,934, 18,492 to 18,497, and 19,506 to 19,511, respectively, can be recognised and transcleaved by the 3C-like proteinase. Five mature products with sizes of approximately 100 kDa, 65 kDa, 63 kDa, 42 kDa and 35 kDa are released from the ORF1b polyprotein by the 3C-like proteinase-mediated cleavage at these positions. Meanwhile, expression of plasmids containing only the ORF1b region showed no autocleavage of the polyprotein encoded, suggesting that the 3C-like proteinase may be the sole proteinase involved in processing of the 1b polyprotein. These data may therefore represent a complete processing map of the polyprotein encoded by ORF1b of mRNA1.

Deletion mutagenesis has been used to identify essential regions for rescue of coronavirus defective RNAs (D-RNAs). Using this technique on a cloned IBV D-RNA CD-91, we have identified a region potentially important in its rescue. Comparing the sequence of D-RNAs rescued with those not rescued we have deduced that a 72 base region corresponding to base number 13824 to 13896 in the viral genome is required for rescue. This may be an IBV D-RNA packaging signal or a cis-acting element involved in replication. Further experiments and modification of our techniques will be required to differentiate between the two processes.

Robotic exploration of the Martian surface will provide important scientific data on planetary climate, life history, and geologic resources.

We report on the development of new mobile robots for Mars exploration missions. These 'lightweight survivable rover (LSR)' systems are of potential interest to both space and terrestrial applications, and are distinguished from more conventional designs by their use of new composite materials, collapsible running gear, integrated thermal-structural chassis, and other mechanical features enabling improved mobility and environmental robustness at reduced mass, volume, and power. Our first demonstrated such rover architecture, LSR-1, introduces running gear based on 2D composite struts and 3D machined composite joints, a novel collapsible hybrid composite-aluminum wheel design, a unit-body structural- thermal chassis with improved internal temperature isolation and stabilization, and a spot-pushbroom laser/CCD sensor enabling accurate, fast hazard detection and terrain mapping. LSR-1 is an approximately .7 $MIL 1.0 meter(Lambda) 2(W X L) footprint six-wheel (20 cm dia.) rocker-bogie geometry vehicle of approximately 30 cm ground clearance, weighing only 7 kilograms with an onboard .3 kilogram multi-spectral imager and spectroscopic photometer. By comparison, NASA/JPL's recently flown Mars Pathfinder rover Sojourner is an 11+ kilogram flight experiment (carrying a 1 kg APXS instrument) having approximately .45 X .6 meter(Lambda) 2(WXL) footprint and 15 cm ground clearance, and about half the warm electronics enclosure (WEE) volume with twice the diurnal temperature swing (-40 to +40 degrees Celsius) of LSR- 1 in nominal Mars environments. We are also developing a new, smaller 5 kilogram class LSR-type vehicle for Mars sample return -- the travel to, localization of, pick-up, and transport back to an Earth return ascent vehicle of a sample cache collected by earlier science missions. This sample retrieval rover R&D prototype has a completely collapsible mobility system enabling rover stowage to approximately 25% operational volume, as well an actively articulated axle, allowing changeable pose of the wheel strut geometry for improved transverse and manipulation characteristics.

We report a significant advance in space robotics design based on innovation of 3D composite structures and piezoelectronic actuation.

Managed care which emphasizes the rationalization of the way health care is financed, the devolution movement which shifts greater program responsibility to the state and local levels, and the restructuring of Medicaid along the lines of a block grant program, are issues presenting providers of aging services with a daunting set of challenges and opportunities. Providers of aging services will need to build on existing strengths, while developing new strategies, to succeed under an environment of managed care and state and local funding control.

A region of the infectious bronchitis virus (IBV) genome between nucleotide positions 8693 and 10927 which encodes the predicted 3C-like proteinase (3CLP) domain and several potential cleavage sites has been clones into a T7 transcription vector. In vitro translation of synthetic transcripts generated from this plasmid was not accompanied by detectable processing activity of the nascent polypeptide unless the translation was carried out in the presence of microsomal membrane preparations. The processed products so obtained closely resembled in size those expected from cleavage at predicted glutamine-serine (Q/S) dipeptides and included a protein with a size of 35 kDa (p35) that corresponds to the predicted size of 3CLP. Efficient processing was dependent on the presence of membranes during translation; processing was found to occur when microsomes were added posttranslationally, but only after extended periods of incubation. C-terminal deletion analysis of the encoded polyprotein fragment revealed that cleavage activity was dependent on the presence of most but not all of the downstream and adjacent hydrophobic region MP2. Dysfunctional mutagenesis of the putative active-site cysteine residue of 3CLP to either serine or alanine resulted in polypeptides that were impaired for processing, while mutagenesis at the predicted Q/S release sites implicated them in the release of the p35 protein. Processed products of the wild-type protein were active in trans cleavage assays, which were used to demonstrate that the IBV 3CLP is sensitive to inhibition by both serine and cysteine protease class-specific inhibitors. These data reveal the identity of the IBV 3C-like proteinase, which exhibits characteristics in common with the 3C proteinases of picornaviruses.

In order to investigate the mechanisms involved in the processing of infectious bronchitis virus polyproteins, several candidate regions of the genome have been cloned and expressed in vitro. During these studies it was observed that the translation product encoded by one of these clones (pKT205) was poorly expressed. Biochemical and genetic analyses revealed that the basis for the poor expression was a post-translational event involving ubiquitination of the protein and degradation by an ATP-dependent system operating in the reticulocyte lysate used for the in vitro expression. Two independently acting regions which conferred instability were identified, one of which mapped to the predicted 3C protease domain, contained within the 5′ end of the clone, while the other, more C-terminal region, was effective in conferring instability upon a heterologous protein to which it had been transferred. These regions may influence the stability of the authentic viral protein(s) in vivo and hence allow for the control of their expression and/or function at the level of proteolysis by cellular protease(s).

Airbags have become an increasingly accepted automobile safety feature that can reduce the morbidity and mortality associated with motor vehicle collisions. We present the following case report of an unusual fatality with multiple internal injuries from a minor mechanism motor vehicle collision. The cause of injuries was determined to be secondary to the deployment of a driver's side airbag without the concomitant use of a lap-shoulder belt.

The gene encoding the spike (S) protein from two geographically distinct strains (American and British) of canine coronavirus (CCV) was cloned and sequenced. The nucleotide sequence revealed open reading frames of 1443 or 1453 amino acids, respectively. Structural features include an N-terminal hydrophobic signal sequence, a hydrophilic cysteine-rich cluster near the C-terminus, two heptad repeats and 29 or 33 potential N-glycosylation sites. Pairwise comparisons of S amino acid sequences from these isolates with other CCV strains (Insavc1 and K378) revealed that heterogeneity, found mostly in the form of conservative substitutions, is distributed throughout the canine sequences. However, 5 variable regions could be identified. Similar analysis with feline, porcine, murine, chicken and human coronavirus sequences revealed that the canine sequences are much more closely related to the feline S protein sequence than to the porcine S protein sequences even though they are all from the same antigenic group. Moreover, the sequence similarity between CCV isolates and the feline coronavirus, feline infectious peritonitis virus (FIPV) was comparable. Expression of the CCV or the transmissible gastroenteritis virus (TGEV) S gene using the vaccinia virus system produced a protein of the expected size which could induce extensive syncytia formation in infected canine A72 cells.

Intracerebral inoculation of the neurotropic murine picornavirus, Theiler's murine encephalomyelitis virus (TMEV), results either in an acute encephalitis (GDVII strain) or in the establishment of a persistent infection with the development of demyelinating lesions (BeAn strain). In this article, the expression of the viral RNA polymerase was studied in the central nervous system of both acutely and persistently infected mice and in infected cells in tissue culture. Similar numbers of acutely infected glial cells (80–85%) expressed both viral polymerase and structural proteins in vitro while a much smaller proportion of persistently infected glial cells (0·6–0·9%) expressed these proteins. Following infection of mice with GDVII, many cells in the brain were found to express polymerase. However, in the spinal cord of mice persistently infected with BeAn, very few cells were found to express the polymerase while many more cells showed the presence of viral structural proteins. This suggests that a restriction in viral replication, possibly at the level of polymerase expression, may be a feature of the persistent infection. However, enough polymerase was expressed to maintain a polymerase-specific antibody response in a number of infected animals as late as 21 months post-infection. Mechanisms that may be involved in the establishment and maintainance of TMEV persistence are discussed with reference to these findings.