Metkephamid, a systemically active analog of methionine enkephalin with potent opioid alpha-receptor activity.

Metkephamid is an analog of methionine enkephalin that retains high affinity for the delta receptor and is a systemically active analgesic. Since it is at least 100 times more potent than morphine as an analgesic when placed directly into the lateral ventricles, and is 30 to 100 times more potent on the delta receptor and yet is roughly equipotent on the mu receptor in vitro, it is concluded that it probably produces analgesia by action on delta receptors as well as, or rather than, on mu receptors. It has less tendency to produce respiratory depression, tolerance, and physical dependence than standard analgesics, and it is presently undergoing clinical trial.

Chronic granulomatous disease and McLeod syndrome in a black child.

A 3-year-old black male child with X-linked chronic granulomatous disease and red cells of the rare McLeod phenotype is presented. The red cells showed acanthocytosis and did not react with anti-KL. Similarly the leukocytes were nonreactive with anti-Kx. The Xk and Xg linkage could not be investigated since all members of his family were Xg (a+).

The role of an adherent cell in the production of monocyte chemotactic factor.

The functional role of an adherent cell in assisting a variety of in vitro immune responses is well established. An assay for human monocyte chemotaxis in vitro was utilized as a means of investigating the role of adherent cells in the production of the monocyte chemotactic factor that is produced by peripheral blood lymphocytes upon stimulation with Concanavalin A. Depletion of a population of adherent cells by passage of peripheral blood mononuclear cells through a Sephadex G-10 column rendered the latter incapable of producing monocyte chemotactic factor. The requirement for adherent cells in the production of a "lymphokine" is in agreement with many previous works of a similar nature performed in other experimental systems.

Using a knowledge, attitudes and practices survey to supplement findings of an outbreak investigation: cholera prevention measures during the 1991 epidemic in Peru.

BACKGROUND: To assess the effectiveness of the cholera prevention activities of the Peruvian Ministry of Health, we conducted a knowledge, attitudes, and practices (KAP) survey in urban and rural Amazon communities during the cholera epidemic in 1991. METHODS: We surveyed heads of 67 urban and 61 rural households to determine diarrhoea rates, sources of cholera prevention information, and knowledge, attitudes, and practices regarding ten cholera prevention measures. RESULTS: Twenty-five per cent of 482 urban and 11% of 454 rural household members had diarrhoea during the first 3-4 months of the epidemic. Exposure to mass media education was greater in urban areas, and education through interpersonal communication was more prevalent in rural villages. Ninety-three per cent of rural and 67% of urban respondents believed they could prevent cholera. The mean numbers of correct responses to ten knowledge questions were 7.8 for urban and 8.2 for rural respondents. Practices lagged behind knowledge and attitudes (mean correct response to ten possible: urban 4.9, rural 4.6). Seventy-five per cent of respondents drank untreated water and 91% ate unwashed produce, both of which were identified as cholera risk factors in a concurrently conducted case-control study. CONCLUSIONS: The cholera prevention campaign successfully educated respondents, but did not cause many to adopt preventive behaviours. Direct interpersonal education by community-based personnel may enhance the likelihood of translating education into changes in health behaviours. Knowledge, attitudes, and practices surveys conducted with case-control studies during an epidemic can be an effective method of refining education/control programmes.

PIP: The authors conducted a knowledge, attitudes, and practices (KAP) survey in urban and rural Amazon communities during the 1991 cholera epidemic to assess the effectiveness of the Peruvian Ministry of Health's cholera prevention activities. Diarrhea rates, sources of cholera prevention information, and knowledge, attitudes, and practices regarding 10 cholera prevention measures were determined by surveying the heads of 67 urban and 61 rural households. 25% of 482 urban and 11% of 454 rural household members had diarrhea during the first 3-4 months of the epidemic. Exposure to mass media education was greater in urban areas, while education through interpersonal communication prevailed in rural villages. 93% of rural and 67% of urban respondents believed they could prevent cholera. Rural respondents were slightly more knowledgeable than urban respondents about cholera. Overall, however, practices did not reflect their knowledge and attitudes; 75% of respondents drank untreated water and 91% ate unwashed produce.

Solid-phase indirect immunogold localization of human sperm antigens reacting with antisperm antibodies in human sera.

A simple solid-phase indirect immunogold assay for light microscopic localization of the binding of sperm-reactive antibodies in human sera to surface-associated antigenic sites on human sperm is described. Antigen-antibody interaction is localized by using a gold-tagged second antibody directed against the first antibody. Silver enhancement of the 5 nm gold particles makes the immunologic reaction visible by light microscopy. An ELISA was used to identify serum samples as positive or negative for sperm-reactive antibodies. These serum samples were blind-coded and were used to develop and to evaluate this solid-phase indirect immunogold assay. When results obtained by this immunogold assay for the detection of sperm-reactive antibodies were compared with those obtained by the ELISA, there was agreement in 82 of the 100 sera tested. As a localization assay, this indirect immunogold assay is simple to perform and enables stable, high resolution light microscopic localization of the pattern of antigenic sites on sperm surface membranes.

Effects of androgen deprivation on the histology of adult chimpanzee testis.

Until primate sperm are exposed to the unique microenvironment of the epididymis, they are not capable of fertilization or vigorous motility. Many of the proteins that contribute to the unique microenvironment of the primate epididymis, and thus to sperm maturation, are dependent on androgens to induce their synthesis and secretion. GnRH antagonists have proved effective in suppressing LH and testosterone synthesis and secretion, and thus in maintaining a state of androgen deprivation or functional hypogonadotropism. We report here the effects of GnRH antagonist-induced androgen-deprivation on the histology of the testicular interstitium and seminiferous epithelium of the adult male chimpanzee. After only 21 days of androgen-deprivation, chimpanzee testicular tissues exhibit specific atrophic changes, including the loss of contact between developing spermatocytes and between Sertoli cells and their developing spermatids, alterations in cell development resulting in missing maturation steps (elongating Sc and structurally complete Sd2 spermatids) and inappropriate cell associations, varying degrees of cytoplasmic degradation in germ cells, Sertoli cells, and Leydig cells, and a tubular lumen obscured by masses of sloughed primary and secondary spermatocytes and what appear histologically to be Sb1 and Sd1 spermatids.

Histological effects of androgen deprivation on the adult chimpanzee epididymis.

Primate sperm acquire functional maturity, including vigorous forward motility and the ability to fertilize an ovum, as they transit the unique, regional microenvironment of the epididymal lumen. Several proteins secreted into this luminal fluid are epididymal-specific and androgen-dependent, and thus contribute potentially to sperm maturation. For the adult male chimpanzee, we report the effects of GnRH antagonist-induced androgen deprivation on the histology of the epithelia and interstitium composing the ductuli efferentes, ductus epididymis, proximal ductus (vas) deferens. After 21 days of androgen deprivation, epididymal tissues exhibit characteristic atrophic changes, including cellular disorganization, degradation, and loss of structures. Androgen-deprived cytoplasm is differentially and characteristically disrupted, vacuolated, and reduced in volume, resulting in decreased epithelial height and loss of stereocilia. Most principal cell nuclei appear hyperchromatic, smaller in size, more irregular in outline, and disordered in arrangement, while others appear swollen and vacuolated. Apical cells of the efferent ducts and the basal cells and microvillar borders of the ductus epididymis seem minimally affected by androgen deprivation. Such histologically differential responses suggest correspondingly that androgen is differentially essential to the maintenance of the epididymis and thus to normal functioning of the component tissues. Therefore, epididymal epithelia directly and their secretions indirectly are differentially androgen-dependent.

Sequential histology of the adult chimpanzee epididymis.

The chimpanzee epididymis consists of three gross anatomical regions: caput (Cp), corpus (Cr), and cauda (Cd). During epididymal transit, chimpanzee sperm undergo maturational changes (affecting motility, capacitation, and the acrosome reaction), which enable fertilization. Elucidation of the sequential histology of the chimpanzee epididymis is preliminary to determining the role of each region of epididymal duct in the process of sperm maturation. We report the histology, in the adult male chimpanzee, of sequential segments of epididymis from the proximal efferent ductules through the Cp, Cr, and Cd regions of the epididymis proper to the proximal ductus deferens. These data illustrate the gross architecture of the tissue and reveal the histology characteristic of the diverse epithelia composing the epididymal ducts at each level. The tubules composing the epididymis of the adult male chimpanzee present at least 16 histologically-distinct epithelia and their transitional forms. Such diversity of epithelia suggests a corresponding diversity of function. Although there are some clear differences, the histological appearance of the ducts, and of the component epithelia of the chimpanzee epididymis, is remarkably similar to that reported for the human.

Germ cell maturation and cellular associations in the seminiferous epithelial cycle of the chimpanzee.

Seminiferous tubule architecture, germ cell maturation steps and cellular associations (stages) of the spermatogenic cycle of the chimpanzee (Pan troglodytes) are resolved. Cross sections of seminiferous tubules usually exhibit 2 to 4 stages, occasionally 1, and rarely 5; stages are not functionally sequential in structurally contiguous regions. The cellular maturation steps are: dark type A stem cell (Ad), pale type A (Ap), type B(B) spermatogonia; resting or preleptotene (P1), leptotene (L), zygotene (Z), pachytene (P), diplotene (Di) primary spermatocytes; meiotic divisions (M1, M2); secondary spermatocytes (2 degrees S); six developmental stages of the spermatid (Sa, Sb1, Sb2, Sc, Sd1, Sd2) composing spermiogenesis. The germ cell maturation steps characteristic of the six cellular associations (stages I-VI) are: Ad, Ap, B, P, Sa, Sd1 (I); Ad, Ap, B, PI, P, Sa, Sd2 (II); Ad, Ap, B, PI, L, P, Sb1 (III); Ad, Ap, PI, L, P, Sb2 (IV); Ad, Ap, L, Z, P, Di, Sc (V); Ad, Ap, B, Z, P, Di, 2 degrees S, Sc (VI). Surgical pressure trauma causes sloughing of some 2 degrees S spermatocytes and some Sa, Sb1, Sb2, Sd1, and Sd2 spermatids, resulting in missing generations, and disrupts Sertoli cell attachments, affecting germ cell development and associations. In structure and function, chimpanzee spermatogenesis appears most similar to the human.

Immunohistochemical localization of epididymal secretory glycoprotein EP1 in the adult male chimpanzee.

Proteins, synthesized by the epididymal epithelium, are secreted sequentially into the lumen of the ducts epididymis where they effect sperm maturation and enable functional motility and fertilizing capacity. EP1 is a major secretory glycoprotein of chimpanzee (Pan troglodytes) epididymis. The epididymal duct exhibits diverse histology (Smithwick & Young, 1997). Epithelia I-V of the efferent ducts show no characteristic anti-EP1 binding. The densest granules of anti-EP1 reaction product appear in epithelium VI adjacent to the basal lamina in the infranuclear region of the principal cells (PCs), in the cytoplasm of the apical half of the PCs, and in the perinuclear and perivacuolar cytoplasm of the basal cells. In epithelia VII-XIV of the ductus epididymis proper, anti-EP1 binding decreases distally and is localized in the cytoplasm of the PCs and basal cells, among the stereocilia of the luminal border, within various microvillar borders, and in the luminal fluid. Therefore, EP1 appears to be synthesized and secreted primarily in the caput region of the ductus epididymis and may be reabsorbed nonselectively across epithelia with apical microvilli, including the non-ciliated cells of efferent ducts, the distal corpus and cauda of the ductus epididymis, and the proximal ductus deferens.

Estimate of epididymal transit time in the chimpanzee.

During their passage through the epididymis, sperm undergo functional changes which result in their maturation and in their ability to fertilize ova. Maturational changes effected during epididymal transport are attributable to sequential changes in various regions of the plasmalemma of the sperm head and flagella. These functional changes in the plasmalemma result, at least in part, from the sequential binding of proteins secreted by the epididymal epithelium into the epididymal lumen. An estimate of epididymal transit time is essential to such investigations. Time elapsed from a testicular arterial infusion of a single pulse of tritiated-thymidine to the release of 3H-labeled sperm into the lumen of the seminiferous tubule was about 39 days. Seminal fluid-free 3H-labeled sperm first appear in the ejaculate about 41 +/- 1 days post-infusion. Total transit time for 3H-labeled Sd2 sperm released into the tubular lumen to appear in the ejaculate is estimated as the difference between these values. Since total transit time is equal to the seminiferous tubule transit time plus the epididymal transit time, epididymal transit time constitutes some lesser portion of the total transit time of 2 +/- 1 days.

Duration of spermatogenesis and relative frequency of each stage in the seminiferous epithelial cycle of the chimpanzee.

To determine the duration of 1 spermatogenic cycle, a single pulse of tritiated thymidine was infused into a branch of the spermatic artery in each of 3 chimpanzees (Pan troglodytes). Samples were recovered surgically prior to infusion, at 1 h, and at 3, 8, 14, 16, 17, 28, 30, 33, 40, 44, and 48 days postinfusion. Tissues were fixed in Bouin's solution, dehydrated, paraffin-embedded, sectioned at 5 micrometers, and stained. Pre-infusion samples were used in morphometric studies to estimate the percentage frequency of area occupied by each of the 6 cellular associations (stages I-VI) characteristic of chimpanzee spermatogenesis, and thus, to estimate the days duration of each stage. To estimate the duration of 1 spermatogenic cycle, pre- and post-infusion, tissue sections were coated with undiluted Kodak NTB2 liquid autoradiographic emulsion and incubated at 4 +/- 1 degree C. At optimum exposure times, slides were processed with Kodak D-19 and Fixer; light microscopic analyses were conducted to determine the most mature labeled cell in stage III for each of the sample times. The duration of the 6 stages (I-VI) are 4.2, 2.0, 4.3, 1.5, 1.3 and 0.6 days, respectively, and the duration of 1 spermatogenic cycle is approximately 14 days. Thus, the duration of spermatogenesis from the Ap spermatogonium to mature Sd2 spermatid is approximately 62.5 +/- 1.5 days or 4.46 spermatogenic cycles.

Effect of liquefaction time on semen parameters in chimpanzee (Pan troglodytes) ejaculates.

Ejaculates were obtained from eight adult male chimpanzees trained to use an artificial vagina. Spontaneously liquefied fractions were collected at 0-1, 1-2 and 2-4 h after ejaculation. The mean volumes of the spontaneously liquefied fractions did not differ significantly among the three fractions. The total number of sperm, the percentage of motile sperm and the percentage of live sperm decreased significantly (P < 0.005) between the 0-1 and 1-2 h fractions. Citrate concentration and alpha-glucosidase activity (mU/ml) decreased significantly (P < 0.005) from 0-1 to 1-2 to 2-4 h fractions. The volume of the liquefied fraction correlated positively (P < 0.001) with total protein and with total citrate at 0-1, 1-2 and 2-4 h. The total number of sperm correlated positively total with total alpha-glucosidase activity at 0-1, 1-2 and 2-4 h. Among the biochemical parameters measured, total protein and total citrate correlated positively at 0-1, 1-2 and 2-4 h. © 1995 Wiley-Liss, Inc.

Characteristics of chimpanzee (Pan troglodytes) ejaculates collected by rectal probe electrostimulation and by artificial vagina.

This study compares characteristics of ejaculates collected from 16 adult male chimpanzees using rectal probe electrostimulation (RPE) and from 10 adult male chimpanzees trained to use an artificial vagina (AV). Ejaculate weight, semen volume, and sperm number were significantly lower (P < 0.01) and percentage liquefaction was significantly higher (P < 0.01) in ejaculates collected by RPE. Percentages of motile sperm and of live sperm in semen did not differ significantly between the two collection methods. Total amounts of protein and of α-glucosidase activity were significantly lower (P < 0.01) in seminal fluid from RPE samples. For ejaculates collected by RPE, semen volume correlated positively with protein (r = 0.8640, P < 0.001), fructose (r = 0.6976, P < 0.001), and citrate (r = 0.6976, P < 0.001); sperm number correlated positively with α-glucosidase activity (r = 0.6547, P < 0.001); and protein correlated positively with fructose (r = 0.5906, P < 0.002), citrate (r = 0.5926, P < 0.002) and α-glucosidase activity (r = 0.6006, P < 0.001). For ejaculates collected by AV, semen volume correlated positively with percentage liquefaction (r = 0.6058, P < 0.001), protein (r = 0.8055, P < 0.001), fructose (r = 0.6606, P < 0.001), and citrate (r = 0.8272, P < 0.001); sperm number correlated positively with percentage of motile sperm (r = 0.4196, P 0.004); percentage of motile sperm correlated positively with percentage of live sperm (r = 0.4388, P < 0.002); and, protein correlated positively with fructose (r = 0.6947, P < 0.002) and with citrate (r = 0.5926, P < 0.002). These data show that there is a significant difference in semen parameters and in biochemical parameters of ejaculates obtained by RPE and by AV. © 1995 Wiley-Liss, Inc.

Sodium, potassium, and protein concentrations and 2D-SDS-page of epididymal luminal and ejaculated seminal fluids of the adult chimpanzee (Pan troglodytes).

The concentration of soluble protein and of sodium and potassium ions was estimated in chimpanzee caput epididymal luminal fluid, cauda epididymal luminal fluid, and ejaculated seminal fluid. Protein concentration was 48.5 ± 1.5 µg/µl in caput fluid, 26.8 ± 2.0 µg/µl in cauda fluid, and 53.0 ± 7.9 µg/µl in seminal fluid. Sodium concentration was 127.0 ± 7.0 mM in caput fluid, 34.5 ± 1.8 mM in cauda fluid, and 18.8 ± 1.8 mM in seminal fluid. Potassium concentration was 58.0 ± 0.0 mM in caput fluid, 56.8 ± 5.2 mM in cauda fluid, and 77.0 ± 1.7 mM in seminal fluid. Proteins in caput epididymal, cauda epididymal, and ejaculated seminal fluids, with approximate molecular weights (kDa) between <14.4 and 45.0 kDa and apparent isoelectric points (pIs) between 4.5 and 7.5, were resolved by two-dimensional SDS-polyacrylamide gel electrophoresis (2D-SDS-PAGE) and silver stained. In caput fluid, the most intensely stained polypeptides resolved between 14.4 and 21.5 kDa (pI 5.4-7.4). In cauda fluid, the number and intensity of stained components increased markedly, and the most intensely stained polypeptides resolved between 21.5 and 31.0 kDa (pI 5.7-7.3). In seminal fluid, polypeptides between <14.4 and 45.0 kDa (pI 5.7-7.4) appeared characteristically diffuse and distributed. These results demonstrate that the ions and the polypeptides in the luminal microenvironment change significantly along the epididymal duct of the male chimpanzee. © 1994 Wiley-Liss, Inc.