Geo. F. Brooks

Microbiology and pathogenesis of acute salpingitis as determined by laparoscopy: What is the appropriate site to sample?☆

Acute salpingitis is a polymicrobial disease. Neisseria gonorrhoeae and anaerobic gram-positive cocci were the predominant microorganisms isolated from the fallopian tubes of salpingitis patients. Gonococci were isolated from the fallopian tubes in eight of 35 (23%) patients; anaerobic bacteria were recovered from 10 of 35 (28.5%). Although Chlamydia trachomatis was not recovered from the fallopian tube exudate, there was abundant serologic evidence of chlamydial infection in the salpingitis patients. Twenty-three percent of patients with paired sera had a fourfold rise in IgM and IgG titer, which was consistent with systemic chlamydial infection. Comparison of cultures obtained via laparoscopy and culdocentesis suggested that culdocentesis is not an accurate reflection of the microbial milieu in the fallopian tube.

Immunogenicity and evolutionary variability of epitopes within IgA1 protease from serogroup A Neisseria meningitidis

Five murine epitopes were defined and mapped within IgA1 protease produced by Neisseria meningitidis. Epitopes 1 and 2 were present in IgA1 protease from all strains, and from Neisseria gonorrhoeae. Epitopes 3 through to 5 varied between subgroups of serogroup A meningococci. but have remained constant over decades within the subgroups, except for epitope 4, which changed between 1983 and 1987 during the spread of subgroup III meningococci from Asia to Africa. Binding of monoclonal antibodies to epitopes 1, 4 and 5 neutralized enzymatic function. Human sera containing antibodies to lgA1 protease as a result of natural infection inhibited binding of monoclonal antibodies to epitope 4 but not to the other epitopes.

CEACAM is not necessary for Neisseria gonorrhoeae to adhere to and invade female genital epithelial cells.

Neisseria gonorrhoeae has a repertoire of up to 11 opacity‐associated (Opa) proteins that are adhesins. Most Opa proteins adhere to CEACAM antigens and when CEACAM molecules are present on the surface of transfected epithelial cells their binding by Opa is thought to induce invasion of these cells by gonococci. In this study, we investigated whether several malignant epithelial cell lines, normal cervical and fallopian tube epithelial cell cultures, as well as normal fallopian tube tissue express several of the CEACAM molecules, and whether gonococci use these molecules for adherence and invasion of these female genital epithelial cells. A primary cervical cell culture and metastatic cervical cell line ME180 both expressed CEACAM as shown by whole cell ELISA and flow cytometry, and increased the surface expression of total CEACAM during incubation with Opa+ gonococci. Opa+ gonococci both adhered to and invaded these cells; CEACAM‐specific monoclonal antibody (MAb) partially abolished this interaction. Two primary fallopian epithelial tube cell cultures, a primary cervical cell culture and two malignant cell lines, HEC‐1‐B and HeLa, did not express CEACAM nor was CEACAM mRNA present. No evidence of either intracellular or secreted extracellular CEACAM was found with HEC‐1‐B and HeLa cells. Opa+ gonococci both adhered to and invaded CEACAM non‐expressing cells; however, Opa+ gonococcal association with these non‐expressing cell lines could not be inhibited with CEACAM‐specific MAb. These data show that CEACAM is not always expressed on female genital epithelial cells and is not essential for gonococcal adherence and invasion. However, when CEACAM is expressed, Opa+ gonococci exploit it for the adherence to and invasion of these cells.

Opa (protein II) influences gonococcal organization in colonies, surface appearance, size and attachment to human fallopian tube tissues

Opa-expressing variants of Neisseria gonorrhoeae strain F62-SF and an Opa- variant, all non-piliated, were examined for differences in the interaction of the bacteria within colonies and in attachment to and damage of human fallopian tube mucosa. Expression of certain Opas was associated with the formation of transparent colonies where the bacteria were tightly packed and evenly spaced within the colonies. Expression of other Opas was associated with the formation of opaque colonies where the gonococci were less tightly packed and were unevenly spaced. Distinct differences in the size of the gonococci and in their surface characteristics were dependent upon the Opa being expressed. Certain Opas were associated with gonococci that had significantly larger cross-sectional areas and bigger perimeters. Scanning electron microscopy showed that OpaC- and OpaD-containing variants yielded greater mucosal damage than OpaB-containing and Opa- variants with the least damage caused by the OpaA-containing variant (clumped bacteria from dark opaque friable colonies). The mucosal damage after 60 min incubation included shortening and decreased numbers of microvilli on non-ciliated cells and invagination and sloughing of ciliated cells. Differences in the interactions of gonococci within colonies and in attachment to fallopian tube mucosa and damage to the mucosal cells occurred with different Opa-expressing variants of N. gonorrhoeae strain F62-SF.

Clindamycin-associated hepatotoxicity

Abstract Hepatotoxicity associated with intravenous clindamycin phosphate therapy is reported in a drug abuser cured of staphylococcal endocarditis. Coincident with the administration of large doses of clindamycin, marked hepatic enzyme abnormalities were noted, and liver biopsy showed lobular disruption, pseudogranulomas, hepatocyte necrosis, eosinophilic bodies and mononuclear cell infiltration. Cessation of clindamycin therapy resulted in return of liver enzymes to normal, and a second liver biopsy performed 15 days later showed improvement. Reports of clindamycin-associated hepatic enzyme changes are reviewed; further observation for clindamycin-associated hepatotoxicity is warranted.

Vancomycin-induced neutropenia complicating bone marrow recovery in a patient with leukemia. Case report and review of the literature

Neutropenia associated with intravenous vancomycin therapy is reported in a patient with chronic myelogenous leukemia. The patient received 12 days of vancomycin therapy without incidence; however, a second course of vancomycin initiated on hospital day 14 produced severe neutropenia. This delayed onset is typical of vancomycin-induced neutropenia. The neutropenia reversed, without complications, as soon as the vancomycin was discontinued.

Expression of gonococcal protein II in Escherichia coli by translational fusion

Aprotein II (P.II) gene from Neisseria gonorrhoeae was cloned in Escherichia coli and characterized by DNA sequence analysis. As with other reported P.II sequences, this gene contains an ATG initiation codon which is out of frame with respect to the remainder of the P.II amino acid sequence. A translational fusion was constructed in E. coli which linked the P.II sequence to the signal peptide of β‐lactamase. This P.II fusion differs from the gonococcal protein only in the first seven residues at the N terminus. In E. coli, the P.II fusion product exhibits properties analogous to those of P.II in N. gonorrhoeae. The P.II fusion product is a major component of the E. coli outer membrane and it is exposed on the cell surface. The P.II fusion protein also exhibits the heat‐modifiable phenotype of gonococcal P.II.

Clinical use of the new beta-lactam antimicrobial drugs. Practical considerations for physicians, microbiology laboratories, pharmacists, and formulary committees.

New beta-lactam antimicrobial agents with extended antibacterial activity for gram-negative bacilli are being developed and marketed. These drugs provide major advances, especially for treatment of serious infections caused by multiresistant organisms. Several of the drugs have been marketed and many more will be available. Some of these drugs are considerably more costly than the older beta-lactams. The large number of new antimicrobial drugs coupled with their high costs pose complex problems for physicians, microbiology laboratories, and pharmacists. Community hospitals, large general hospitals, and tertiary care hospitals have different needs for patient care and will need different formats for unbiased education, susceptibility testing, pharmacy stocking, and controlling or monitoring for inappropriate use.

Detection of vancomycin-intermediate Staphylococcus aureus with the updated Trek-Sensititre System and the MicroScan System. Comparison with results from the conventional Etest and CLSI standardized MIC methods.

Vancomycin-intermediate Staphylococcus aureus (VISA) organisms have minimum inhibitory concentrations (MICs) of 4 to 8 microg/mL and are often associated with vancomycin treatment failure. Detection of VISA has remained problematic. A comparison of 4 methods to detect VISA was done. Of the 20 VISA isolates, the Clinical and Laboratory Standards Institute (CLSI) broth microdilution method yielded susceptible end points of 2 microg/mL for 7, MicroScan (Siemens Healthcare Diagnostics, West Sacramento, CA) for 2, Trek Sensititre (Trek Diagnostic Systems, Cleveland, OH) for 1, and Etest (AB Biodisk North America, Piscataway, NJ) for none. Comparison with the CLSI method showed essential agreement for 95% or more for the Etest, MicroScan, and Trek methods; categorical agreement was as follows: Etest, 60%; MicroScan, 65%; and Trek, 60%. Reliance on a single automated method for determining vancomycin MICs could lead to misclassification of some VISA isolates as vancomycin susceptible. At least 2 methods, including the Etest, should be used when confirming VISA because of slight differences in results from different methods around the end points of 2 and 4 microg/mL .

Prevalence of gene sequences coding for hypervariable regions of Opa (protein II) in Neisseria gonorrhoeae

Opas (protein IIs) are a family of surface‐exposed proteins of Neisseria gonorrhoeae. Each strain of N. gonorrhoeae has multiple (10–11) genes encoding for Opas. Identifiable elements in opa genes include the coding repeat within the signal sequence, conserved 5′ and 3′ regions, and hypervariable regions (HV1 and HV2) located within the structural gene. N. gonorrhoeae strains appear to have many biological properties in common that are either HV‐region‐mediated or associated with the presence of specific HV regions, suggesting that HV regions could be found in many clinical isolates. Oligonucleotides from three source strains representing three conserved regions of opa, 12 HV1 regions, and 14 HV2 regions were used by dot blot analysis to probe 120 clinicial isolates of N. gonorrhoeae. The probe for the coding repeat hybridized to all 120 strains, the 3′ conserved‐region probe reacted with 98% of the strains, and the 5′ conserved‐region probe with 90% of the strains. Nine HV1 probes hybridized to 3.3–39.2% of the strains, and 13 of the HV2 probes hybridized to 1.7–25% of the isolates. Analysis of the number of probes that hybridized to each of the isolates showed that 19% did not hybridize with any of the HV1 probes and 25% did not hybridize with any of the HV2 probes. Approximately three‐quarters of the isolates hybridized with one, two or three of the HV1 probes or one, two or three of the HV2 probes; 89% of the isolates hybridized to least one HV1 or oe HV2 probe. The data indicate that some genes encoding HV regions of N. gonorhoeae Opa proteins are widely distributed in nature.