Cynthia Smith

Cortical auditory dysfunction in benign rolandic epilepsy

Purpose: To evaluate cortical auditory function, including speech recognition, in children with benign rolandic epilepsy (BRE).

A pharmaceutical profile of diabetic patients

Although there have been innumerable studies documenting various aspects of the morbidity caused by diabetes mellitus in the population, very little attention has been paid to patterns of therapeutic management of diabetic patients. The United States Public Health Service Ambulatory Care Data System (USPHS ACDS), a computerized patient information entry and recording system with a complete pharmaceutical record for each patient, was used to compare patterns of pharmaceutical consumption among diabetic patients as opposed to non-diabetic patients in a population of approximately 90,000 individuals. Drug use by diabetics was significantly higher than by non-diabetics. Cardiovascular drug use, in particular, was considerably higher. Substantially higher consumption of anti-lipemic agents, anti-gout drugs, anti-hypertensives, sedatives and tranquilizers was also found in the diabetic population. The higher use of all drugs by diabetics could be partially explained by a demonstrably higher frequency of out-patient visits by diabetics. However this factor alone could not account for very much higher use of certain selective drug groups by the diabetics. In most cases, these selective increases among the diabetics paralleled expected patterns of disease for which those drug groups are prescribed. The investigation of pharmacotherapeutic profiles of the diabetic population adds a new dimension to the epidemiological study of this disease.

ATP and SH3 Binding Sites in the Protein Kinase of the Large Subunit of Herpes Simplex Virus Type 2 of Ribonucleotide Reductase (ICP10)

The large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) is a multifunctional protein. It consists of a ribonucleotide reductase and a serine/threonine protein kinase (PK) domain, which has three proline-rich motifs consistent with SH3-binding sites at positions 140, 149, and 396. We used site-directed mutagenesis to identify amino acids required for kinase activity and interaction with signaling proteins. Mutation of Lys176 or Lys259 reduced PK activity (5-8-fold) and binding of the 14C-labeled ATP analog ρ-fluorosulfonylbenzoyl 5′-adenosine (FSBA) but did not abrogate them. Enzymatic activity and FSBA binding were abrogated by mutation of both Lys residues, suggesting that either one can bind ATP. Mutation of Glu209 (PK catalytic motif III) virtually abrogated kinase activity in the presence of Mg2+ or Mn2+ ions, suggesting that Glu209 functions in ion-dependent PK activity. ICP10 bound the adaptor protein Grb2 in vitro. Mutation of the ICP10 proline-rich motifs at positions 396 and 149 reduced Grb2 binding 20- and 2-fold, respectively. Binding was abrogated by mutation of both motifs. Grb2 binding to wild type ICP10 was competed by a peptide for the Grb2 C-terminal SH3 motif, indicating that it involves the Grb2 C-terminal SH3.

The Herpesvirus hypothesis--are Koch's postulates satisfied?

Abstract The evidence associating herpes simplex virus type 2 (HSV-2) with squamous cervical carcinoma is critically reviewed. Thirteen criteria for a causal relationship between a virus and a human cancer are listed and evidence is presented demonstrating that in the case of HSV-2 and cervical cancer these have been fulfilled. The original Koch postulates are discussed from the standpoint of modern concepts of virology. In this context, emphasis is placed on the identification in cervical tumor cells of the viral protein ICP 10/AG-4. This protein is expressed in HSV-2-transformed hamster cells and its expression correlates well with the oncogenic potential of the transformed lines. Consistent with these findings, the expression of ICP 10/AG-4 in cervical cells and the prevalence of the specific antibody that it induces, reflect the progression of the cervical tumor.

Proteins of herpesvirus type 2. Isolation and immunologic characterization of two viral proteins in a virus-specific antigenic fraction

Abstract Previous reports from our laboratory have shown that “crude” AG-e, a type-commons HSV antigen, induces an antigen-driven cell-mediated immune response in patients with HSV cervicitis or cervix cancer. Anti-crude AG-e sera give rise in crossed immunoelectrophoresis against total viral soluble antigenic mixtures (HSV-2 (G) SAM) to one precipitin band (“pure” AG-e). Pure AG-e immunologically identical to crude AG-e as determined by immunodiffusion and it resolves into two infected cell proteins, ICP 12 (MW: 140,000) and ICP 14 (MW: 130,000) upon SDS-acrylamide gel electrophoresis. ICP 12 and ICP 14 are purified to radiochemical homogeneity by SDS-acrylamide gel electrophoresis. Antisera to ICP 12 react with HSV-2 (G) SAM in crossed immunoelectrophoresis and both anti-ICP 12 and ICP 14 sera stain HSV-2 (G) infected cells. ICP 12and ICP 14 appear to be envelope proteins as evidenced by: (i) the absorption of the reactivity of anti-ICP 12 and ICP 14 sera with HSV-2 (G) virions but not with “mock-virus” preparations, (ii) the ability of the anti-ICP 12 and ICP 14 sera to neutralize HSV-2 in presence of anti-IgG, and (iii) the resolution of a protein with the relative mobility of ICP 14 in virions the surface of which was iodinated with lactoperoxidase.

Expression and cellular compartmentalization of a herpes simplex virus type 2 protein (ICP 10) in productively infected and cervical tumor cells.

Antiserum to ICP 10, a herpes simplex virus type 2 (HSV-2) protein that is expressed in cells neoplastically transformed by viral DNA sequences within the Bgl II/Hpa I CD fragment, specifically precipitates the ICP 10 protein from HSV-2 infected cells and stains cells infected with HSV-2 for 4 to 16 hrs by indirect immunofluorescence. At 4 hr post infection (p.i.), the staining is primarily perinuclear, while at 16 hr p.i., it is cytoplasmic and intranuclear. Compartmentalization studies indicate that the 35S-[L]-methionine labeled ICP 10 is detectable in both the cytoplasmic and nuclear fractions early and late in infection. However, in its phosphorylated form, ICP 10 is undetectable in the nuclear fraction late in the viral reproductive cycle. Anti-ICP 10 serum stains a high (75%-83%) proportion of cervical tissue with pathological findings of dysplasia or carcinoma, as well as atypical exfoliated cells from these patients. Cervical tumor tissue from 4 of 12 patients also stains with antiserum to another purified viral protein complex designated ICP 12/14. In the majority of atypical cells with mild or moderate changes, ICP 10 localizes in the cytoplasm, while the majority of atypical cells with severe changes also display nuclear staining with anti-ICP 10 serum. While exfoliated atypical cells from 60% of patients with dysplasia are positive for ICP 10, those from only one half of these patients stain also with anti-ICP 12/14 serum and this staining is strictly cytoplasmic. Atypical cells from three patients in these series stain with the anti-HSV-2 serum but are negative for both ICP 10 and ICP 12/14. Exfoliated atypical cells from patients with CIS or invasive cancer stain equally well with all three antisera.