Sexual development in Plasmodium berghei: the use of mitomycin C to separate infective gametocytes in vivo and ookinetes in vitro.
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Sixty-five Holstein bull calves were used to study the effects of unilateral castration (UC) and increased plane of nutrition on the growth and development of the reproductive system. Bulls were slaughtered at 1 wk., 2, 4, 8 and 16 months. Half of each slaughter group above one week was unilaterally castrated at 7 days of age. Half of the bulls remaining at 6 months of age received 90% of their recommended daily TDN allowance while the remainder received 120%. Compensatory hypertrophy was evident as early as 2 months and the degree of compensation increased for the duration of the experiment (Age x UC, P<.01). By 16 months of age the remaining testis of UC animals was 73% heavier than the average testis weight of intact bulls. While epididymal weight was significantly increased by UC, seminal vesicle weight was not. UC bulls produced significantly more sperm per testis than intact bulls both from the onset of puberty to slaughter and for the 16 week period prior to slaughter. Testis sperm concentration was similar in UC and intact bulls. UC at one weel of age caused greater testis growth and greater sperm production per testis, but did not promote earlier puberty.
Sixty-five Holstein bull calves were assigned in an experiment to determine the effects of unilateral castration (UC) at 1 week of age and of two levels of nutrition after 6 months on reproductive development to 16 months. Five animals were killed at 1 wk and half the remainder UC at that age. Groups of 5 in all factorial groups were killed at 2, 4, 8 and 16 months. Tubular diameter increased with age (P<.01) and at 8 and 16 months with UC (P<.01). Epithelial area at 8 and 16 months increased with age and UC (P<.01). The percents of tubular and intertubular tissue varied with age (P<.01) with the tubular tissue having its highest value at 8 months. Indexes of both total tubular and intertubular tissue were increased with age and UC (P<.01). The number of type A spermatogonia per cross section of stage 1 tubules of 16-month bulls was increased by UC (P<.05).
Pituitary gonadotropic hormones were assayed in 65 Holstein bulls from 7 days to 16 months. Pituitary LH concentration and content at 2, 4, 8 and 16 months increased (P<.01) with age, while FSH content increased with age (P<.01) but was lower in UC bulls at 2, 4 and 8 months and higher at 16 months (A x UC, P<.01) as compared to intact bulls. In five samples of plasma collected at 90-minute intervals, one day each month from 1 to 15 months in 10 of the bulls killed at 16 months, LH concentration and variance changed (P<.01) with age reaching maxima at 4 and 3 months respectively. Plasma testosterone concentration and variance changed (P<.01) with age, reaching maxima at 10 and 9 months respectively. In the other 10 bulls killed at 16 months, assays of plasma collected before and after exposure to a teaser showed that stimulation increased LH by 20 minutes after exposure but LH declined by 60 minutes while testosterone was increased 20 (P<.05) and 60 (P<.01) minutes after exposure.
A total of 155 male buffalo calves were classified into 17 groups according to chronological age. The body weight was recorded on a balance or computed. Sera samples were analysed for thyroxine (T(4)) and triiodothyronine (T(3)) by radioimmunoassay. Highest concentration of thyroid hormones (T(4), 87.6+/-17.6; T(3), 3.1+/-0.07 ng/ml) were seen during the first week after birth followed by a gradual decline until two months of age. Later, the mean T(4) and T(3) levels fluctuated between 30 and 40 ng/ml and at around 1.0 ng/ml, respectively, except for a mild peak at 12 to 15 months of age. T4:T3 ratio did not vary significantly among various age groups. Both T4 and T3 were negatively correlated with age.
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DNA contents of individual stages of Plasmodium berghei were measured by direct microfluorometry after Feulgen-pararosaniline (SO2) staining. Sporozoites, intra-erythrocytic ringforms and trophozoites (until at least 15 h after invasion) are haploid and non-synthesizing DNA. DNA is synthesized just before and during schizogony, which takes 4-6 h. Genome duplication and segregation are alternating events throughout this process. Mature micro- and macrogametocytes have DNA contents between the haploid and diploid value; most, if not all of the DNA in excess of the haploid value is synthesized during the last 5-10 h of maturation. During gametogenesis microgametocytes within 8-10 min synthesize DNA steadily and at a very high rate to more than the octoploid value while the DNA content of macrogametocytes remains constant. Fertilization in vitro takes place within 1 h after gamete formation. Within 2 h and coinciding with the onset of meiosis the zygote then synthesizes DNA up to almost the tetraploid value, after which synthesis stops during ookinete development. All the above mentioned processes of DNA synthesis are reversibly inhibited by aphidicolin (C50 from 3-13 microM). From the rate of DNA synthesis during microgametogenesis we calculated a minimum of 1300 origins of replication in the haploid genome of P. berghei.
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Clitoral measurements were obtained in normal children; small increases in clitoral dimensions were associated with growth and puberty. Patients with androgen excess had major increases in clitoral size, clearly different from normal values.
Among 30 children with hepatic ductular hypoplasia and normal extrahepatic bile ducts, 15 formed a hemogeneous, readily recognizable group. In addition to chronic cholestasis, they have characteristic facies, a mesosystolic murmur, vertebral arch defects, growth retardation, mental retardation, and hypogonadism. Typical biochemical and histologic features aid in differentiation of this group from patients with other varieties of biliary disease.
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