A simple method for immune analysis of proteins following sodium dodecyl sulphate polyacrylamide gel electrophoresis.

Immune analysis of polypeptides separated by SDS-PAGE provides a technique useful in the characterisation of complex mixtures. We describe a simple procedure involving immune analysis of fractionated PAGE gels. Polypeptides of purified herpesvirus simplex, HSV 'excreted antigen' and cell culture 'control' preparations were separated by SDS-PAGE. Following electrophoresis, protein was eluted from gels, adsorbed to plastic and analysed using an RIA technique. When using a mouse anti-HSV1 serum, 7 peaks of immune reactivity were observed with pure virus and 6 with HSV 'excreted antigen'. Only 1 peak of reactivity was observed with a 'control' antigen.

Virucidal activity of chlorhexidine on strains of Herpesvirus hominis, poliovirus, and adenovirus.

The infectivity titre of two strains of Herpesvirus hominis was reduced by not more than six log(10) units by 90 minutes' exposure to chlorhexidine at a final concentration of 0.02% at room temperature. Under identical conditions the compound was ineffective against a vaccine strain of poliovirus and against an adenovirus. Experimental conditions are described whereby the toxic effect of the disinfectant on the tissue culture assay systems are minimized.

Isolation of a Coxsackie virus group B, type 5, from the heart of a fatal case of myocarditis in an adult.

The cause of a fatal case of myocarditis in an adolescent male was established at necropsy by the isolation of Coxsackie virus group B, type 5, from myocardium and bowel. An antibody assay on a single serum is reported. This is believed to be a first report and its significance is assessed.

A prospective study of cytomegalovirus and herpes simplex virus disease in renal transplant recipients.

A prospective study of 84 renal graft recipients demonstrated cytomegalovirus (CMV) disease after transplantation in 37% of patients. Reactivation infection was found in 20 of 44 patients (46%) who were seropositive for CMV prior to transplant and primary CMV disease occurred in 11 of 40 (28%) initially seronegative patients. Nearly all cases of primary disease (91%) were associated with symptoms and in these cases CMV was probably acquired via the donated kidneys. Only 35% of the reactivation infections were associated with clinical symptoms. Actuarial life tables indicated that CMV disease did not reduce the length of graft survival. Herpes simplex virus (HSV) infections were diagnosed in 44 (52%) of the patients and included a fatal case of disseminated disease associated with hepatitis.

Human papillomavirus (HPV) genotypes associated with laryngeal papilloma.

Biopsy specimens from 14 patients treated for laryngeal papillomatosis were tested for the presence of human papillomavirus (HPV) genome by the technique of DNA-DNA hybridisation. According to the age of initial presentation, cases were subdivided into juvenile (less than 16 years) and adult onset (older than 16 years) groups. Histological investigation confirmed that it was impossible to distinguish the groups on this basis. Molecular virology using both dot blot and Southern transfer techniques showed that 10 cases carried the HPV type 6 genome, three cases HPV type 11, and in one case no HPV DNA was detected. All six adult onset cases carried HPV 6 sequences while the juvenile onset group comprised four HPV 6 and three HPV 11 cases. In the juvenile onset group more females were affected; in the adult onset group more males were affected. Two of the patients shown to have HPV type 11 sequences in their biopsy material were the most resistant to treatment. One of the adult onset cases subsequently developed a squamous cell carcinoma of the larynx in which HPV 6 DNA was detected. As far as we know this is first time that HPV-DNA has been confirmed in laryngeal papilloma undergoing malignant change.

Disseminated herpes simplex virus infection with hepatitis in an adult renal transplant recipient.

A rare occurrence of disseminated herpes simplex virus infection with hepatitis in an adult renal transplant recipient is described.

Polymerase chain reaction for diagnosis and identification of distinct variants of Crimean-Congo hemorrhagic fever virus in the United Arab Emirates.

Viral hemorrhagic fever has re-emerged in the United Arab Emirates (UAE) since November 1993. Genomic RNA of Crimean-Congo hemorrhagic virus (C-CHFV) was detected by a newly developed, nested reverse transcriptase polymerase chain reaction (RT-PCR) in the sera of four (25.0%) of 16 suspected cases of viral hemorrhagic fever. The RT-PCR was based on oligonucleotide primers deducted from the small RNA segment encoding the nucleoprotein of the virus. By comparison with a nucleotide sequence of a C-CHFV isolate from a Chinese sheep, a divergence of 10.0-11.8% was detected in the C-CHFV variants causing the UAE outbreak. In the four positive sera, three phylogenetically distinct C-CHFV variants were amplified and confirmed by direct sequencing of the PCR fragments. These C-CHFV sequences were obtained directly from sera of infected humans without prior propagation in cell culture. The RT-PCR allows rapid detection of genomic C-CHFV RNA in clinical specimens and study of the molecular epidemiology of this infection.

A rat model of herpes encephalitis with special reference to its potential for the development of diagnostic brain imaging.

A rat model of herpes encephalitis using intraocular inoculation of herpes simplex virus strain SC16 was investigated. Virus distribution in the brain was examined by virus isolation and immunocytochemical staining using immuno-gold silver and peroxidase-anti-peroxidase. At 5 days post-infection virus was found in the thalamus, hypothalamus, septum, colliculus, geniculate bodies, the pons, trapezoidium and medulla oblongata, but less frequently, in the cerebellum and occipital lobes. Possible routes of spread of virus and the potential of this model in neuro-radiological scanning procedures are discussed.

The treatment of herpes encephalitis.

The advent of acyclovir has accentuated the problem of diagnosis and particularly the need for brain biopsy in patients with suspected herpes simplex encephalitis. U.S.A. and U.K. practices are contrasted and the current U.K. study of acyclovir in herpes simplex encephalitis is mentioned in which retrospective and relatively non-invasive methods of diagnosis are employed.

Meningo-encephalitis caused by Coxsackievirus group B type 2. Diagnosis confirmed by measuring intrathecal antibody.

We describe a case of meningo-encephalitis caused by Coxsackievirus group B type 2. Diagnosis was suggested by isolation of virus from faeces and high titres of serum antibody in which Coxsackie virus B2 specific IgM antibody was detected. Evidence of intrathecal production of antibody confirmed the diagnosis.

Characterization of two rotaviruses differing in their in vitro and in vivo virulence.

The proteins, genomic RNA and disassembly conditions and pathogenesis in vivo for a virulent mouse rotavirus (MRV) and a less virulent bovine rotavirus (BRV) were compared. An obvious difference in the mobility of several genomic RNA segments were observed in one-dimensional gels. Reassortants obtained by replacement of gene 4 in BRV with MRV gene 4 indicated that the dose of the virus used and the clinical outcome in vivo was determined by gene segment 4. Under in vitro conditions, a comparison of the inner capsid proteins by partial proteolytic peptide mapping did not reveal any difference between corresponding proteins. However, this technique did identify differences between the two corresponding outer capsid proteins of these viruses. These differences, in turn, may account for the increased stability of MRV, as compared to BRV, when subjected to calcium-chelating and chaotropic agents and may be one of the mechanisms involved in conferring virulence on the virus. The observed variability between the 84K proteins (VP4) provided a basis for further investigations in order to locate a potential virulence determinant, since it has been observed by us and others that the gene coding for this protein plays a role in determining virulence. A comparison of the carboxypeptidase digests of the MRV and BRV VP4 revealed an area of variability between amino acids 307 and 407, which may represent the site of a virulence determinant.

Influenzavirus A2 infections presenting with febril convulsions and gastrointestinal symptoms in young children.

We present the clinical features of Influenzavirus A2 infection in 75 young children admitted to a children's hospital. The most common presenting features were febrile convulsions. vomiting, coughing, diarrhea, and anorexia. At any age, the illness may present with respiratory tract symptoms and signs but in young babies gastroinestinal symptoms are often the presenting complaint. Children aged one to three years often present with febrile convulsions. Only in older children does the adult pattern begin to emerge. Evidence is put forward to suggest that encephalitis in association with influenza can be due to direct invasion of the central nervous system by the virus.

Persistence in herpes simplex virus infections.

Diseases of man caused by the virus of herpes simplex fall into two broad categories. The primary disease occurs only once in any individual's life and is caused by transmission of virus from an already infected human. Thereafter, the individual may be subject to recurrent herpetic disease, the manifestations of which are different from the primary disease. Recurrent disease varies in severity from trivial, to incapacitating and frankly lethal (as in diseases resulting from the virus's neurotropic and oncogenic properties). The source of the virus in recurrent herpetic disease has never been conclusively resolved, but is almost certainly endogenous to the patient. Theories, case reports and experiments exist to show that endogenous virus may, in periods of clinical quiescence, be latent (or persistent) at the site of the recurrent lesions itself, or more remotely in nerve tissues related to the site of recurrence.

Herpes encephalitis.

Four cases of herpes encephalitis treated with dexamethasone and cytarabine. Cases were all verified by brain biopsy and immunofluorescent techniques. Attention is called to the need for early diagnosis, but the difficulty of deciding when to perform brain biopsy is discussed. Early treatment with effective drugs is necessary to prevent permanent brain damage.

Herpes simplex infections in atopic eczema.

One hundred and seventy nine children with atopic eczema were studied prospectively for two and three quarter years; the mean period of observation being 18 months. Ten children had initial infections with herpes simplex. Four children, very ill with a persistently high fever despite intravenous antibiotics and rectal aspirin, continued to produce vesicles and were given intravenous acyclovir. There were 11 recurrences among five patients. In two patients the recurrences were as severe as the initial lesions, and one of these children had IgG2 deficiency. Use of topical corticosteroids preceded the episode of herpes in only three of the 21 episodes. Symptomatic herpes simplex infections are common in children with atopic eczema, and are suggested by the presence of vesicles or by infected eczema which does not respond to antibiotic treatment. Virological investigations are simple and rapid: electron microscopy takes minutes, and cultures are often positive within 24 hours.