Peroxidase in human cervical mucus during the menstrual cycle

Blain, J. A.; Heald, P. J.; Mack, A.; Shaw, C.E.L.

doi:10.1016/0010-7824(75)90064-5

Cited by 15 publications

(7 citation statements)

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“…The microbicidal activity of H202 is considerably increased by the enzyme peroxidase in the presence of a halide ion (14). Among the peroxidases which can function in this way are those of milk and saliva (lactoperoxidase) (24), neutrophils and monocytes (myeloperoxidase) (14,16), eosinophils (eosinophil peroxidase) (13), and genital tract secretions (rat uterine fluid and human cervical mucus) (3,18). The peroxidase and the halide ion greatly enhance the H202-dependent microbial antagonism between two bacterial species (15), between bacteria and viruses (17), between bacteria and spermatozoa (19), and between bacteria and mammalian cells (4).…”

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confidence: 99%

Prevalence of hydrogen peroxide-producing Lactobacillus species in normal women and women with bacterial vaginosis

et al. 1989

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A predominance of Lactobacillus species in the vaginal flora is considered normal. In women with bacterial vaginosis, the prevalence and concentrations of intravaginal Gardnerella vaginalis and anaerobes are increased, whereas the prevalence of intravaginal Lactobacillus species is decreased. Because some lactobacilli are known to produce hydrogen peroxide (H2O2), which can be toxic to organisms that produce little or no H2O2-scavenging enzymes (e.g., catalase), we postulated that an absence of H2O2-producing Lactobacillus species could allow an overgrowth of catalase-negative organisms, such as those found among women with bacterial vaginosis. In this study, H2O2-producing facultative Lactobacillus species were found in the vaginas of 27 (96%) of 28 normal women and 4 (6%) of 67 women with bacterial vaginosis (P less than 0.001). Anaerobic Lactobacillus species (which do not produce hydrogen peroxide) were isolated from 24 (36%) of 67 women with bacterial vaginosis and 1 (4%) of 28 normal women (P less than 0.001). The production of H2O2 by Lactobacillus species may represent a nonspecific antimicrobial defense mechanism of the normal vaginal ecosystem.

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Prevalence of hydrogen peroxide-producing Lactobacillus species in normal women and women with bacterial vaginosis

et al. 1989

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“…Removal of endogenous peroxidase. The peroxidase enzyme was normally within mucus that closely adhered to cells from the female genitalia and was also contained within lymphocytes infiltrating the infected area (3,22). Due to the nature of the enzyme system used in our studies, during initial experiments this endogenous peroxidase led to a high background stain of uninfected cells, causing false-positive reactions or difficult interpretations of PXAPX results.…”

Section: +mentioning

confidence: 94%

“…Samples to be counterstained by the PAP (17) were processed as follows: distilled water, 10 dips; distilled water, 5 min; Gill's hematoxylin (Polysciences, Inc., Warrington, Pa.) single strength (1 part Gill's no. 3 triple strength plus 2 parts 25% ethylene glycol in water), 2 min; distilled water, 10 dips; distilled water, 10 dips; 70% ethanol, 10 dips; 0.5% concentrated HCI in 70% ethanol, 1 dip to decolorize to salmon color; 70% ethanol, 10 dips; 70% ethanol, 10 dips; 3.0% concentrated ammonium hydroxide in 70% ethanol, 1 dip to a blue color; 70% ethanol, 10 dips; 70% ethanol, 10 dips; 95% ethanol, 5 dips; 95% ethanol, 2 min; Gill's modified OG-6 (Polysciences, Inc.), 2 min; 95% ethanol, 10 dips; 95% ethanol, 10 dips; Gill's modified EA (Polysciences, Inc.), 2.5 min; 95% ethanol, 10 dips; 95% ethanol, 10 dips; absolute ethanol, 10 dips; xylene, 10 dips; xylene, 5 min; Permount and cover slip. Controls for all specimens consisted of: (i) identical treatment of Cytospin preparations as just described (PXAPX or PXAPX-PAP) with rabbit preimmunization serum at appropriate dilutions used in place of immune antiserum; (ii) Cytospin preparations which received only the PAP stain; and (iii) HSV-1or HSV-2-infected Vero cells on cover slips to serve as positive controls in the PXAPX or PXAPX-PAP procedure.…”

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confidence: 99%

Detection of herpes simplex virus infection of female genitalia by the peroxidase-antiperoxidase method alone or in conjunction with the Papanicolaou stain

Pearson¹,

Fleagle²,

Docherty³

1979

J Clin Microbiol

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The sensitive peroxidase-antiperoxidase (PXAPX) method individually and in conjunction with the Papanicolaou (PAP) stain was used to detect herpes simplex virus (HSV) in specimens from human female genitalia. Initial studies using a model system of HSV-1or HSV-2-infected Vero cells established (i) acetone as the most effective fixative, (ii) optimal dilutions of preimmunization and anti-HSV serum for differentiation of infected from noninfected cells, (iii) optimal concentration of 3,3'-diaminobenzidine tetrahydrochloride and H202 for maximal staining of infected cells with minimal background reaction, and (iv) removal of endogenous peroxidase with absolute MeOH. These various parameters, once established, were utilized in the PXAPX or PXAPX-PAP on human specimens from the vulva or cervix. In these specimens, examined by standard light microscopy, PXAPX-positive cells were dark brown with a single nucleus or multiple nuclei. By coupling the PAP to the PXAPX, detailed nuclear observations of PXAPX-positive cells were possible and revealed nuclear changes consistent with HSV infection, including syncytium formation, chromatin condensation, and an occasional Cowdry type A inclusion. The PXAPX and PXAPX-PAP correlated (r = 0.742) over a period of 72 h with HSV isolation from these samples. MATERIALS AND METHODS Virus. Virus stocks of HSV-1 (strain 69-85) and HSV-2 (strain 316-D) were obtained from F.

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“…The effectiveness of using the that the decrement in light scatter intensity caused by immunoperoxidase technique on human specimens, the addition of a n absorbing dye can be enhanced by however, is compromised by the endogenous peroxidase restricting the collection of scattered light to the 3"-6" activity commonly found in mammalian mucosecretions angular interval (9). In the present study, we adapted (5) and inflammatory cells (181, and this activity results in nonspecific staining that can confuse diagnostic interpretation. Specimens are therefore frequently pretreated with Or Other reagents to inactivate endogenous peroxidase (18)(19)(20)23), but such pretreat-Medical Center, Chicago, IL 60611.…”

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confidence: 99%

Flow cytometric detection of herpes simplex virus type 2 infected and transformed cells by immunoenzymatic and by indirect immunofluorescence staining

1988

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The glucose oxidase antiglucose oxidase (GAG) immunoenzymatic staining procedure has been used to detect herpes simplex virus (HSV) antigens microscopically. In this study, the GAG procedure was adapted to cells in suspension, and its potential usefulness in flow cytometry was examined. HSV‐2 infected monkey kidney and HSV‐2 transformed mouse cells were stained using antisera to HSV‐2 or to an HSV‐2 specific protein with a molecular weight of 38 Kd, respectively, with the GAG procedure. Flow cytometric analysis of the GAG stained cells was then performed by the measurement of scattered light intensity in the angular intervals l°−2°, 2.5°−19°, and 3°−6°. The greatest scattered light intensity decrement caused by staining occurred in the 3°−6° angular interval, as predicted by previous work. In infected cells, which stain intensely by immunofluorescence, the difference between positively and negatively stained cells was adequate for detecting infected cells using the GAG method; however, this was not the case for the lightly staining transformed cells. The indirect immunofluorescence method of analysis of the same populations was superior to the scattered light method of analysis of the GAG stained infected and transformed cells.

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Peroxidase in human cervical mucus during the menstrual cycle

Cited by 15 publications

References 1 publication

Prevalence of hydrogen peroxide-producing Lactobacillus species in normal women and women with bacterial vaginosis

Prevalence of hydrogen peroxide-producing Lactobacillus species in normal women and women with bacterial vaginosis

Detection of herpes simplex virus infection of female genitalia by the peroxidase-antiperoxidase method alone or in conjunction with the Papanicolaou stain

Flow cytometric detection of herpes simplex virus type 2 infected and transformed cells by immunoenzymatic and by indirect immunofluorescence staining

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