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D F Katz

Publications and source records attributed to D F Katz.

At least 19 recordsLinked to original sources

Methods for assessing rat sperm motility.

Computer-assisted sperm analysis (CASA) systems are becoming more widely used. With this spread of technology come more data from toxicology studies, designed to determine if treatment with putative toxicants affects sperm motion parameters. While these CASA methods provide us with more ways to evaluate toxicity and thus perhaps increase our chances of successfully protecting human health, there is also a greater likelihood that different laboratories will use different methods of collecting data on sperm motility. Different systems used with different methods in different laboratories will inevitably generate data that are difficult to compare. In a prospective attempt to address this issue of comparability and limit the problems, a group of individuals using CASA systems to analyze rat sperm motility convened to discuss methodologic issues, share data, and try to reach a consensus about methods for performing these studies. This article shares those meetings and data in the hope that common methods will enhance interlaboratory comparisons.

Animals

Laboratory methods for assessing human semen in epidemiologic studies: a consensus report.

It is clear that additional methodologic work needs to be performed. Some data gaps described above are being actively investigated. Other standards were not addressed at this meeting; statistical handling of the data, differences among CASA machines, and factors to consider as potential confounders in analysis are just a few. These may be the subject of future workshops, which will also review progress made in the existing knowledge base. For now, this effort represents a first attempt to share information and to use it to encourage investigators in different laboratories to employ similar methods. In this way more direct comparisons among studies can be made, and our collective data base can be strengthened.

Computers

The relation of computer-based measures of sperm morphology and motility to male infertility.

We investigated the relation between various sperm characteristics, including morphometric parameters, and impaired fertility among 596 men who participated in a national study. Semen was collected and processed by using a standardized protocol, and sperm measurements were made using a computer-aided sperm analysis instrument. We defined infertility in two ways: (1) the inability to father a child after trying for a year or longer, and (2) the number of children fathered. We found that all measures of sperm motion were decreased among men with impaired fertility. After adjustment for the other motion parameters and various potential confounders, however, only the percentage of progressive cells was associated with infertility. One morphometric parameter, the mean length/width ratio, was consistently associated with both measures of infertility, even after adjustment for potential covariates. This measure was also strongly associated with infertility among various subgroups defined by poor sperm concentration, motility, and morphology. The sperm length/width ratio appears to be an important correlate of infertility in males.

Adult

A study of the effect of perchloroethylene exposure on semen quality in dry cleaning workers.

The purpose of this investigation was to determine the effects of perchloroethylene (PCE) exposure on human semen quality. We compared the semen quality of 34 dry cleaners with that of 48 laundry workers. We examined the relationships of 17 semen parameters to expired air levels of PCE and to an index of exposure based on job tasks in the last three months. The average sperm concentration was over 80 million for both dry cleaners and laundry workers, but approximately one-quarter of each group was oligospermic. The overall percentage of abnormal forms was similar for the two groups; however, sperm of dry cleaners were significantly more likely to be round (t = -3.29, p = 0.002) and less likely to be narrow (t = 2.35, p = 0.02) than the sperm of laundry workers. These effects were dose-related to expired air levels and to the exposure index after controlling for potential confounders (e.g., heat exposure). The average percent motile sperm for both groups was slightly over 60%; however, sperm of dry cleaners tended to swim with greater amplitude of lateral head displacement (ALH) than those of laundry workers (t = -1.73, p = 0.09), and level of PCE in expired air was a significant predictor of ALH in the multiple regression model (t = 2.00, p = 0.05). In addition, exposure index was a significant negative predictor of the sperm linearity parameter (t = -2.57, p = 0.01). These results suggest that occupational exposures to PCE can have subtle effects on sperm quality. Additional analyses are required to determine whether these effects are associated with changes in fertility.

Adult

A study of the effect of perchloroethylene exposure on the reproductive outcomes of wives of dry-cleaning workers.

The purpose of this investigation was to compare the reproductive outcomes of wives of men exposed to perchloroethylene in the dry-cleaning industry compared to those of wives of laundry workers. Seventeen female partners of dry cleaners and 32 partners of laundry workers were interviewed. The number of pregnancies and the standardized fertility ratios were similar between the two groups. Wives of dry cleaners did not have higher rates of spontaneous abortions. However, wives of dry cleaners were more than twice as likely to have a history of attempting to become pregnant for more than 12 months or to have sought care for an infertility problem. Cox proportional hazards models indicated that dry-cleaners' wives had half of the per-cycle pregnancy rate of wives of laundry workers, when controlling for other potential confounders (estimated rate ratio of 0.54, 95% C.I. = 0.23, 1.27).

Adult

Species differences in susceptibility to 1,3-dinitrobenzene-induced testicular toxicity and methemoglobinemia.

The testicular toxicity and methemoglobinemia induced by 1,3-dinitrobenzene (1,3-DNB) was compared in two species, the Sprague-Dawley rat and the golden Syrian hamster. A marked difference in susceptibility to both endpoints of toxicity was observed. The hamster showed no testicular lesions at dose levels up to 50 mg/kg whereas, as previously reported by others, damage to rat testicular tubules in later stages of spermatogenesis was readily apparent at a 25 mg/kg dose level. Similarly, administration of 1,3-DNB induced substantially less methemoglobinemia in the hamster than in the rat. For example, at the 25 mg/kg dose level peak levels of methemoglobin in the hamster were 15% compared with 80% in the rat. Mortality in the rat also occurred at lower doses than in the hamster (50 vs 100 mg/kg, respectively). In in vitro studies, the capacity of 1,3-DNB and 1,3-DNB metabolites (nitroaniline, nitroacetanilide, aminoacetanilide, diacetamidobenzene) to induce methemoglobinemia was examined in suspensions of red blood cells obtained from both species. Only 1,3-DNB caused the formation of methemoglobin and rat red blood cells were twice as sensitive as hamster red blood cells. The species difference in susceptibility to both methemoglobinemia and testicular toxicity could indicate differences in 1,3-DNB clearance and/or formation of toxic metabolites. Additional metabolic work is under way. This study demonstrates that the hamster is more resistant than the rat to the testicular lesion and methemoglobinemia induced by 1,3-DNB.

Animals

Human cervical mucus: research update.

Evaluation of cervical mucus is a standard for determining the fertile period in natural family planning. Cervical mucus accepts, filters, prepares, and releases sperm for successful transport to the egg and fertilization. Recent scientific advances provide answers to how the mucus regulates fertility as its physical properties change during the menstrual cycle. Transmission electron microscopy reveals small interstices between mucus macromolecules relative to a sperm head. Thus advancing sperm must push aside or cut through the microstructure. The interstices are largest in the periovulatory phase of the cycle. Small magnetic spheres, comparable with the size of a sperm head, are now being used to study the physical properties of the mucus on the scale of individual sperm.

Cervix Mucus

Evidence for the function of hyperactivated motility in sperm.

After insemination, mammalian sperm undergo a striking change in flagellar beat pattern, termed hyperactivation. In low-viscosity culture medium, nonhyperactivated sperm flagella generate relatively symmetrical, low-amplitude waves, while hyperactivated sperm flagella generate an asymetrical beating pattern that results in nonprogressive movement. Since sperm encounter highly viscous and viscoelastic fluids in the female reproductive tract, the progress of hyperactivated sperm was compared with that of nonhyperactivated and transitional sperm in media of increasing viscosity. Hamster sperm obtained from the caudal epididymis were incubated in a medium that promotes capacitation. After 0, 3, and 4 h of incubation, the majority of the sperm exhibited, respectively, activated, transitional, and hyperactivated motility. At each of these time points, aliquots of sperm were removed from incubation and added to solutions of 0, 5%, 10%, 20%, and 30% Ficoll in medium. Samples containing mostly hyperactivated sperm (4 h) maintained higher swimming and flagellar velocities and were able to generate greater forces in response to increased viscous loading than activated sperm (0 h). Transitional sperm (3 h) showed an intermediate response. The paths of hyperactivated sperm through solutions of 20% and 30% Ficoll were considerably straighter than those made through medium alone. This is the first demonstration that hyperactivation can confer a mechanical advantage upon sperm in the oviduct where they may encounter viscous oviductal fluid and a viscoelastic cumulus matrix.

Animals

The use of a urinary estrone conjugates assay for detection of optimal mating time in the cynomolgus macaque (Macaca fascicularis).

Forty-four female cynomolgus macaques (Macaca fascicularis) were examined to determine the optimum fertile period for mating. Daily urinary estrone conjugates (E1C) were measured, beginning on day 7 of the menstrual cycle, until a 1.5-gold E1C rise above the baseline was detected. The females were bred the next morning. Pregnancies were verified in all animals at day 18 postbreeding, and/or on day 25 postbreeding. Serum progesterone levels were used to correlate the relationship between ovulation and the E1C peak. Forty-four of the 57 cycles indicated a urinary E1C peak between days 10-15 of the menstrual cycle; this peak occurred on the day following the initial 1.5-fold to twofold rise in 90% of the cycles. A single 2-hr mating period the day before, the day of, or the day after the E1C peak resulted in conception in 17 of 44 (38.6%) animals.

Animals

Mechanisms of filtration of morphologically abnormal human sperm by cervical mucus.

It is well known that cervical mucus restricts penetration of morphologically abnormal human sperm, both in vitro and in vivo. However, the mechanisms of such restriction are not well understood. Using videomicrography to simultaneously analyze the motions and morphology of individual human sperm, we analyzed differential penetration of normal and abnormal sperm into fresh human cervical mucus. Abnormal sperm swam slower in mucus than the normal sperm, but their flagellar beat parameters were not commensurately different. Multivariate statistical analysis of the relationship between individual sperm velocity and flagellar beat parameters indicated that the heads of the abnormal sperm experienced greater resistance from the mucus than did normal heads. Differential mucus resistance, more than altered motile vigor, appears to be responsible for the restriction of abnormal sperm during migration through mucus.

Cervix Mucus

Factors regulating mammalian sperm migration through the female reproductive tract and oocyte vestments.

Mechanisms of mammalian sperm migration through the female reproductive tract and ovum vestments are described. The perspective is biophysical as well as biochemical and morphological, and the focus is upon the role of sperm motility in these processes. Sperm forward progression is characterized as an interactive process between the the cell and its environment, and the mediation of flagellar bend propagation by the physical properties of its surroundings is described. These properties, together with flagellar beat kinematics, sperm morphology, and surface properties, determine the magnitude of the forces generated by sperm and their consequent rate of progression. Sperm interactions with the cervical mucus, the cumulus oophorus, and the zona pellucida are described. The poorly understood affinity of the sperm surface for the macromolecules of the mucus, cumulus, and zona is stressed, as is the viscoelastic structural mechanical resistance of these biopolymers to sperm motion. The kinematics and consequences of hyperactivated sperm motion are presented, with emphasis on objective characterization of such motion (as a biomarker), along with analysis of the mechanical advantage that such motion may confer on spermatozoa during egg-vestment interaction.

Animals

Movement of cynomolgus and rhesus monkey spermatozoa collected from the lower female reproductive tract.

Postcoital (pc) cervical mucus was collected in 73 menstrual cycles of cynomolgus monkeys and in 43 cycles of rhesus monkeys at 2, 6, 10, 30 hr pc. Videomicrography was used to analyze sperm numbers and movement in the mucus. Both cynomolgus and rhesus monkeys had comparable populations of motile sperm in the mucus at 2 hr pc. However, by 6 hr pc, cervical mucus from cynomolgus monkeys contained twice as many total sperm and motile sperm as mucus from rhesus monkeys (P less than .05). Mean swimming speeds of the free-swimming cervical sperm were similar for the two species at this time. No motile sperm were recovered in mucus from rhesus monkeys at 30 hr pc. In cynomolgus monkeys, however, 14 of the 26 animals examined at 30 hr pc had motile sperm in their mucus. These sperm exhibited lower percent motility, percent free-swimming sperm, and swimming speed than those sperm observed at 6 hr pc. Uterine sperm were collected by transcervical or transuterine aspiration from cynomolgus monkeys. In the transcervical technique, sperm were successfully obtained in four of nine animals examined at 6 hr and in four of five animals at 30 hr pc. The percentage of motile sperm in the uterine fluid was high, 82% +/- 4%, and the swimming speeds (86 +/- 2 microns/sec) were higher than those observed in cervical mucus. Approximately 5-10% of the uterine sperm exhibited swimming motions similar to the hyperactivated motility seen in most mammals. These findings indicate that the sperm cervical mucus interaction in vivo in cynomolgus monkeys has more similarities to the human situation than does the interaction in rhesus monkeys.

Animals

Semen characteristics of Vietnam veterans.

As part of an epidemiologic study of the health status of a randomly selected group of Vietnam veterans, we measured the semen characteristics of 324 Vietnam veterans and compared them with a similar group of 247 veterans who did not serve in Vietnam. The participation rate was 81% in both groups. Measurements of sperm concentration, movement characteristics, and head dimensions were performed, using the Cellsoft computer-assisted semen analysis system. We found that Vietnam veterans had significantly (p less than 0.05) lowered mean sperm concentrations (64.8 x 10(6) sperm/mL for Vietnam veterans vs 79.8 x 10(6) sperm/mL for non-Vietnam veterans), and Vietnam veterans were twice as likely to have sperm concentrations less than or equal to 20 million/mL (odds ratio = 2.7, 95% confidence interval = 1.3-5.7). Vietnam veterans also had a significantly lowered mean proportion of morphologically normal sperm heads (57.9% vs 60.8%), with significantly longer mean major axis length and head circumference. The proportion of motile cells, velocity, linearity, amplitude of lateral head displacement, and beat frequency were not different between the two groups. Despite differences in sperm characteristics, Vietnam and non-Vietnam veterans reported fathering similar numbers of children.

Adult

Human cervical mucus and its interaction with sperm: a fine-structural view.

Cervical mucus is a glycoprotein gel whose biological functions depend upon its macromolecular architecture. Using freeze-substitution fixation techniques, we have used transmission electron microscopy to examine the fine structural aspects of mucus, before and after unidirectional physical shearing, and during its interaction with sperm. The microstructure of mucus that has not been directionally stretched consists of a homogenous pattern of interconnecting electron-dense elements. The thickness of the primary structural elements varies from 0.04 to 0.5 microns, giving the impression that the elements have fibrillar but also ribbon-like properties. This dimension is smaller than the sizes depicted by prior studies using scanning electron microscopy. Within the mucus interior, the interstitial distance between adjacent primary elements ranges from 0.5 to 0.8 microns, and is filled with a fibrous network of secondary structural elements. This interstitial dimension is also significantly smaller than that suggested by prior electron microscopic work. At the exterior borders of the mucus, the interstitial sizes are reduced. After physical stretching by forceps, the mucus microstructure undergoes a radical deformation. In some specimens, the primary structural elements become longitudinally aligned and are less interconnected, with compaction at the exterior mucus borders. In other specimens, the primary structural elements become laterally compressed along the longitudinal axis; within the regions of compression, the intrapore diameter is reduced to less than 0.1 microns. These compressed regions will often exclude spermatozoa. Individual sperm deform the local mucus microstructure. The mucus directly anterior to the sperm head is stretched, and the mucus adjacent to the bending flagellum is compressed on the forward side and stretched on the opposite side.

Cervix Mucus

Computer-aided sperm analysis (CASA): image digitization and processing.

The active undulations of a sperm flagellum are a basic expression of cell vitality. They reflect a number of intracellular processes, and also reveal the interaction between the sperm and its environment. The pattern of sperm-head motion during locomotion is a hydrodynamic consequence of the behavior of the flagellum. Hence, studies of sperm-head motion should provide insights into cellular function and membrane integrity. Because of its importance in fertility, considerable attention is devoted to the analysis of sperm-head motion at both a clinical and basic biological level. Historically, such work has been done by hand. Recently, new, computer-aided sperm analysis (CASA) systems have become available to automate this process. These new systems promise to advance the study of cell behavior because they facilitate the objective collection of large amounts of data at relatively low cost. At present, however, few workers understand the relative strengths and limitations of such systems. There is a tendency to uncritically accept CASA results because they are computer generated. In this article, we describe how CASA systems work, and discuss several technological and methodological problems which can produce spurious results. We argue for the development of optical, machine, and specimen preparation standards which will facilitate inter-laboratory comparisons of CASA data.

Animals