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Biomedical subjects

S B Ackerman

Publications and source records attributed to S B Ackerman.

11 recordsLinked to original sources

A quick, reliable staining technique for human sperm morphology.

The evaluation of sperm morphology is still an important parameter in the diagnosis of the infertile male. Most techniques used for staining human sperm are very time-consuming. A routine stain used for determining differential count of leucocytes (Diff-Quik stain) was evaluated against the standard Papanicolaou stain. Morphology results from 20 duplicate semen smears using both staining methods were determined separately by 2 technicians using a blind protocol. No significant differences were observed when comparing the two staining methods (paired Student's t-test). The advantages of the Diff-Quik stain are: a) complete staining-to-reading time under 7 min, b) commercially prepared reagents, and c) case of staining procedure.

Humans

Toxicity testing for human in vitro fertilization programs.

Using a mouse embryo culture system, several procedures and materials associated with human in vitro fertilization protocols were tested for potential toxicity. Also, quality-control assays were performed for media prepared by nine different human in vitro fertilization programs. Detrimental effects upon embryo development were observed when culture media were exposed to the following substances: surgical instruments sterilized with Cidex or Cidex-7 or sterilized with ethylene oxide after packaging in Nest Protector Packs, various brands of surgical gloves, and various synthetic materials being evaluated as possible needle or catheter "liners." Results from comparative testing of media and serum supplements prepared by different in vitro programs indicated a wide range in culture medium quality, as assayed by the ability of the reagents to support mouse embryo development. The importance of an animal model system available to all human in vitro programs for routine quality-control analysis and testing of novel uses of materials and innovative methods is discussed.

Animals

Mouse embryo culture for screening in human IVF.

A system for obtaining and culturing mouse two-cell embryos is described, and its importance as a quality control assay and experimental model for human IVF programs is discussed. Over 2000 embryos from B6CBAF1 mice were cultured in either 115 preparations of Ham's F10 medium supplemented with 15% human fetal cord serum (H + FCS, a human IVF medium) or 18 preparations of Krebs' medium (a mouse IVF medium). No significant differences were noted in mean embryo development in the cultures of the two media. After 72 h in culture, the mean % morula + blastocyst development was 93% and 98% for H + FCS and Krebs', respectively. In 8% of the H + FCS cultures at least 75% of mouse embryos failed to grow to either morula or blastocysts by 72 h in culture, and fragmenting embryos were observed in 46% of the H + FCS cultures compared with 0% and 17%, respectively, of Krebs' cultures. Using this system, no significant differences were noted between control media (Krebs' or Ham's F10) and Ham's F12, Menezo's media, or Krebs' medium exposed to small pieces of a medical-grade silicone rubber semen collection device. However, exposure to several types of Vacutainer evacuated blood serum collection tubes did significantly impair the ability of Krebs' medium to support in vitro mouse embryogenesis.

Animals

Probe colorimeter for quantitating enzyme-linked immunosorbent assays and other colorimetric assays performed with microplates.

The performance of a fiberoptic probe colorimeter (model PC800; Brinkmann Instruments, Inc., Westbury, N.Y.) for quantitating enzymatic or colorimetric assays in 96-well microtiter plates was compared with the performances of a spectrophotometer (model 240; Gilford Instrument Laboratories, Inc., Oberlin, Ohio) and a commercially available enzyme immunoassay reader (model MR590; Dynatech Laboratories, Inc., Alexandria, Va.). Alkaline phosphatase-p-nitrophenyl phosphate in 3 M NaOH was used as the chromophore source. Six types of plates were evaluated for use with the probe colorimeter; they generated reproducibility values (100% coefficient of variation) ranging from 91 to 98% when one individual made 24 independent measurements on the same dilution of chromophore on each plate. Eleven individuals each performed 24 measurements with the colorimeter on either a visually light (absorbance of 0.10 at 420 nm) or a dark (absorbance of 0.80 at 420 nm) dilution of chromophore; reproducibilities averaged 87% for the light dilution and 97% for the dark dilution. When one individual measured the same chromophore sample at least 20 times in the colorimeter, in the spectrophotometer or in the enzyme immunoassay reader, reproducibility for each instrument was greater than 99%. Measurements of a dilution series of chromophore in a fixed volume indicated that the optical responses of each instrument were linear in a range of 0.05 to 1.10 absorbance units.

Colorimetry

Effects of Trypanosoma brucei gambiense infections in Microtus montanus on susceptibility to Ehrlich's tumors.

Trypanosoma brucei gambiense infections in the field vole Microtus montanus increased susceptibility to Ehrlich's tumor growth. Whereas uninfected voles were totally resistant to intraperitoneal Ehrlich's ascites tumor cell challenge, over 78% of the animals infected with the trypanosomes developed tumors after challenge. Likewise, when Ehrlich's ascites cells were injected subcutaneously to induce solid tumor formation, only 7% of uninfected controls developed tumors, whereas over 82% of trypanosome-infected animals exhibited malignancies after Ehrlich's cell challenge. Finally, when solid tumors grown in albino CD-1 mice were implanted subcutaneously into uninfected voles, the tumor mass rapidly diminished in size and could not be found when animals were examined 2 weeks postimplant. However, in trypanosome-infected voles, implanted tumors exhibited pronounced expansion, and viable, solid tumors were recovered from over 70% of the challenged voles at 2 weeks postimplant. The implications of trypanosome-induced immunosuppression, especially toward susceptibility to neoplastic growth, are discussed.

Animals

Immunodepression during Trypanosoma brucei gambiense infections in the field vole, Microtus montanus.

The effects of Trypanosoma brucei gambiense infections upon immune responses were examined in an outbred laboratory colony of field voles. Microtus montanus. Antibody levels to challenge with heterologous erythrocytes and bovine serum albumin were significantly depressed in infected animals. Trypanosome infections impaired both primary and secondary humoral responses, although previously established specific antibody levels were not affected by infection. Specific antibody-producing capabilities of previously infected, trypanoicidal drug-treated voles were found to be comparable to uninfected controls, within 3 days following chemotherapy. Cell-mediated hypersensitivity responses to oxazolone were also significantly depressed by trypanosome infection; responses to the initial sensitization of oxazolone and to secondary challenge treatments were depressed compared to uninfected controls. Possible mechanisms of trypanosome-induced immunodepression are discussed.

Animals