Anti-envelope antibody responses in individuals at high risk of hepatitis C virus who resist infection.

Injection drug users uninfected by hepatitis C virus (HCV) despite likely repeated exposure through high-risk behaviour are well documented. Factors preventing infection in these individuals are incompletely understood. Here, we looked for anti-HCV-envelope antibody responses in a cohort of repeatedly exposed but uninfected subjects. Forty-two hepatitis C diagnostic antibody- and RNA-negative injection drug users at high risk of exposure were studied and findings compared to healthy controls and cases with chronic HCV infection. Purified IgGs from sera were tested by ELISA for binding to genotype 1a and 3a envelope glycoproteins E1E2 with further testing for IgG and IgM reactivity against soluble E2. Virus-neutralizing activity was assessed using an HCV pseudoparticle system. Uninfected subjects demonstrated significantly greater IgG and IgM reactivities to envelope glycoproteins than healthy controls with IgG from 6 individuals additionally showing significant neutralization. This study is the first to describe humoral immunological responses targeting the HCV envelope, important for viral neutralization, in exposed uninfected individuals. A subset of these cases also had evidence of viral neutralization via anti-envelope antibodies. In addition to confirming viral exposure, the presence of specific anti-envelope antibodies may be a factor that helps these individuals resist HCV infection.

Elevated interferon-stimulated gene transcription in peripheral blood mononuclear cells occurs in patients infected with genotype 1 but not genotype 3 hepatitis C virus.

Hepatitis C virus (HCV) can be classified into seven distinct genotypes that are associated with differing pathologies and respond differently to antiviral therapy. In the UK, genotype 1 and 3 are present in approximately equal proportions. Chronic infection with HCV genotype 3 is associated with increased liver steatosis and reduced peripheral total cholesterol levels, which potentially influences peripheral immune responses. To understand these differences, we investigated host gene transcription in peripheral blood mononuclear cells by microarray and quantitative PCR in patients with genotype 1 (n = 22) or genotype 3 infection (n = 22) and matched healthy controls (n = 15). Enrichment of genes involved in immune response and inflammatory pathways were present in patients infected with HCV genotype 1; however, no differences in genes involved in lipid or cholesterol metabolism were detected. This genotype-specific induction of genes is unrelated to IL28B genotype or previous treatment failure. Our data support the hypothesis that genotype 1 infection drives a skewed Type I interferon response and provides a foundation for future investigations into the host-pathogen interactions that underlie the genotype-specific clinical outcomes of chronic HCV infection.

The stracathro approach to the hip.

A new approach to the hip joint is described in which the integrity of the gluteus medius is preserved. It provides good exposure of the hip while causing the least disturbance to the abductor mechanism.

Failures of screening and management of congenital dislocation of the hip.

We report the screening of 67,093 infants for congenital dislocation of the hip from 1980 to 1989 and compare the results with those during the preceding two decades. More dislocations have been missed at neonatal examination during the last decade (0.13% of live births). Operative treatment was needed in 54 children (0.08% of live births) some of whom had been diagnosed at birth. We discuss the reasons for the failure of neonatal screening.

A biomechanical study of cast-brace treatment of femoral shaft fractures.

The off-loading characteristics of the cast-braces of 30 patients with fractures of the shaft of the femur have been investigated, during axial loading, using strain-gauge transducers. These were applied at the level of the fracture, where the cast was circumferentially split, and to the hinges of the brace at the knee. They measured the load transferred between the two portions of the thigh cast, and between the thigh cast as a whole and the below-knee cast; by subtraction from the total load on the limb, the skeletal force at the fracture level and at the knee could be calculated. In all patients there was an increase in the fracture load as union progressed which was thought to be due to physiological feedback mechanism from the fracture site. The load carried by the two portions of the thigh cast and by the thigh cast as a whole was proportionately high at first and stabilised at an average of 35 per cent of body weight.

Recurrent osteoid osteoma of the lunate: a case report and review of the literature.

Osteoid osteoma is a benign tumour of bone that rarely localises in the carpal bones. Its treatment by curettage and bone grafting is considered to be curative and its recurrence is thought to be rare. We report a case of an osteoid osteoma of the lunate, which recurred seven years after the initial operation. Recurrent osteoid osteoma of the lunate bone has not been reported in the literature. We present this case report for its atypical presentation and diagnostic difficulty and also to alert the readers of the possibility of an osteoid osteoma as a cause of the chronic unexplained wrist pain in young adults.

Properties of the hepatitis C virus core protein: a structural protein that modulates cellular processes.

The core protein of hepatitis C virus (HCV) is believed to form the capsid shell of virus particles. Maturation of the protein is achieved through cleavage by host cell proteases to give a product of 21 000 MW, which is found in tissue culture systems and sera from infected individuals. However, efficient propagation of the virus is not possible at present in tissue culture. Hence, studies have focused on the properties of the core protein and its possible role in pathologies associated with HCV infection. This review describes key features of the polypeptide and the status of current knowledge on its ability to influence several cellular processes.

The abundance of the herpes simplex virus type 1 UL37 tegument protein in virus particles is closely controlled.

The tegument region of herpes simplex virus type 1 virus particles contains approximately 15 virus-encoded polypeptide species. One of the less abundant species is a 120 kDa protein specified by gene UL37. The abundance of the UL37 protein in infected cells was increased about 20-fold by replacing the native promoter for gene UL37 with the strong immediate early promoter of human cytomegalovirus. This rise in abundance did not induce any detectable increase in the amount of UL37 protein incorporated into virus particles. These data contrast with those previously obtained for a second tegument protein VP22 whose level of incorporation was elevated by increasing its abundance in infected cells. Thus, for the UL37 protein, a mechanism exists to control its abundance in the tegument. Moreover, data obtained with light particles which lack the viral capsid suggest that this controlled incorporation is not directed by interaction with the capsid structure.

Cast bracing of femoral shaft fractures in children: a preliminary report.

This paper reviews the use of cast bracing in fractures of the femoral shaft in children between 1977 at the Royal Aberdeen Children's Hospital. Thirteen children have been treated by cast bracing. All children have achieved a full functional recovery. The average time to cast bracing was 3 weeks, and the average time to discharge was 4 weeks. Thus treated, these children were discharged 4 weeks earlier than those treated by conventional measures. It was found that the technique of cast bracing was applicable to children with no adverse complications.

Organization of the herpes simplex virus type 1 transcription unit encoding two early proteins with molecular weights of 140000 and 40000.

The 5' ends of two early herpes simplex virus type 1 mRNAs have been identified by nuclease S1 and exonuclease VII analysis using cloned virus DNA probes. These mRNAs (5.0 kb and 1.2 kb), located within the genome region between map coordinates 0.56 and 0.60, are unspliced and share a 3' terminus. Genomic DNA at the 5' ends has been sequenced and the 5' termini have been located on the virus DNA sequence. The DNA sequence has revealed signals involved in the initiation of transcription of both mRNAs, and the 5' end of the 1.2 mRNA is encoded within the internal sequences of the 5.0 kb mRNA. The probable translational initiation codons for the polypeptides specified by these mRNAs have been identified, and the results indicate that the coding regions of the two mRNAs do not overlap.

Insertion of DNA sequences at a unique restriction enzyme site engineered for vector purposes into the genome of herpes simplex virus type 1.

We describe the construction of a novel herpes simplex virus (HSV) vector containing a unique XbaI restriction enzyme cloning site in an intergenic position in the short unique genome region. Sequences can be inserted at this site with high efficiency by ligation with XbaI-digested vector DNA. A series of plasmids has been designed for use with this vector. These allow protein coding sequences to be placed under the control of various transcriptional regulation signals and then isolated as XbaI fragments ready for insertion into the vector. The XbaI fragments also contain the beta-galactosidase gene thereby facilitating selection of recombinant virus by screening for blue plaques. A variant of the vector has been made based on the temperature-sensitive (ts) mutant tsK, which expresses only immediate early (IE) genes at non-permissive temperatures. Chloramphenicol acetyltransferase was used as a reporter gene to assess the fidelity of expression of sequences cloned into this position. Under these circumstances IE and early HSV promoters were shown to behave as expected in both wild-type and ts vectors.

Characterization of enveloped tegument structures (L particles) produced by alphaherpesviruses: integrity of the tegument does not depend on the presence of capsid or envelope.

Recent studies have shown that infection with herpes simplex virus type 1 (HSV-1) strain 17 generates in addition to virions a novel type of non-infectious particle. These particles, termed L particles, lack capsids and viral DNA, and consist predominantly of tegument and envelope proteins. We show that L particle production is not restricted to one strain of HSV-1, and that pseudorabies virus and equine herpesvirus type 1 also release particles which are similar in composition to and morphologically indistinguishable from HSV-1 L particles. Data obtained from monoclonal antibody analysis revealed that Vmw175, an immediate early HSV-1 polypeptide which had been previously identified as a virion component, is located predominantly in L particles and not in virions. Following removal of the envelope from L particles, the remaining tegument material largely retained its structural integrity, indicating that the structure of the tegument does not depend on the presence of the capsid or envelope.

Comparing the mortality and morbidity of cemented and uncemented hemiarthroplasties.

A series of 207 consecutive patients admitted to Aberdeen Royal Infirmary over a 1-year period with a diagnosis of displaced subcapital fracture of the neck of the femur were treated by hemiarthroplasty with either a cemented Hasting or an uncemented Monk bipolar prosthesis. Patients were reviewed at an average of 19 months and differences in mortality and morbidity assessed. We found that there was a significant perioperative mortality in the cemented group and no perioperative mortality in the uncemented group. There was no significant difference in the morbidity or complication rate in the two groups, although surviving patients appeared significantly more satisfied with the cemented Hasting prosthesis.

A 3' co-terminus of two early herpes simplex virus type 1 mRNAs.

A 3' co-terminus of two early herpes simplex virus type 1 mRNAs has been identified using the nuclease -S1 mapping procedure with cloned virus DNA probes. These mRNAs (5.0 kb and 1.2 kb), located within the genome region 0.56-0.60, are unspliced and are transcribed rightwards on the prototype genome orientation. The position of their 3' ends has been located on the virus DNA sequence and lies downstream from the polyadenylation signal 5'-AATAAA-3'. This hexanucleotide sequence also was present in the complementary DNA strand and was shown to be the polyadenylation signal for a leftwards-transcribed late mRNA. The abundance within the cytoplasm of the 5.0 kb and 1.2 kb mRNAs was investigated. Results indicated that these mRNAs were regulated in concert. It is suggested that sequences at the 3' co-terminus may be involved in their regulation.

DNA sequence homology between two co-linear loci on the HSV genome which have different transforming abilities.

A transcription unit at the herpes simplex virus (HSV) type 2 transforming region, mtr-2 (map coordinates 0.580-0.625), comprises two early, unspliced mRNAs of 4.5 kb and 1.2 kb which are 3' co-terminal; a region including that specifying the 1.2-kb mRNA has been sequenced. The putative translated portions of these two mRNAs do not overlap and this feature, together with the arrangement of the mRNAs, is similar to the apparently equivalent co-linear HSV-1 locus which however does not transform. A putative stem and loops structure containing a TATA box is located upstream from the 5' terminus of the 1.2-kb mRNA within the translated portion of the 4.5-kb mRNA. Evidence for the generation of this structure by intra-strand reassociation under our hybridisation conditions has been obtained and possibilities are that it may function as a transcription-activated promoter or as an RNA polymerase pause site. A comparison of the equivalent HSV-2 and HSV-1 regions reveals a conserved sequence downstream from the 3' co-terminus which is present at a similar location in many eukaryotic genes (consensus sequence YGTGTTYY). The overall sequence conservation at this transcription unit is high except for regions located at: (1) the untranslated leaders of the 1.2-kb mRNAs; (2) the N termini of the polypeptides specified by the HSV-2/HSV-1 1.2-kb mRNAs; (3) the intergenic region beyond the 3' co termini. Regions (2) and (3) are located within a transforming fragment of HSV-2. The possible significance of these data for HSV-mediated cell transformation is discussed.

A survey of fractures of the proximal femur.

A series of 1494 fractures of the proximal femur, admitted over a 68-month period to the North East Area Accident Unit, is presented. The mortality for the series was 18 per cent, with operative mortality varying according to the fracture site and the method of treatment, being 18.1 per cent for extracapsular fractures, 14 per cent for Smith-Petersen nailing of intracapsular fractures, and 31 per cent for primary prosthetic replacement. In general terms, morbidity and mortality were proportional to age and to pre-existing general medical conditions. The results are presented in full, and are compared with other published series.