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II. evaluation of biological maturation by means of maturity criteria.

Maturation may be conceived of as a series of gradual transformations through time going on in the human body from conception to death as part of the life cycle of the organism. Maturity should be used as the general concept of any specified stage - or level -during the process of maturation. The maturity level can be assessed by examining the attainment of various maturity criteria relating to dental development, skeletal development and pubertal development. In a group of children the attainment can be analyzed by probit analysis using a logarithmic time concept (logarithmic conceptional age). This is exemplified by the attainment of breast stages according to Tanner (10) in the girls of the present study. When examined a child will be rated as being within a certain stage of maturity. The least advanced and the most advanced maturity level consistent with the rating can be established using the probit lines. The relative maturity level is then expressed in SD-scores of the Gaussian fitted distributions of age at attainment of various maturity criteria in a reference group of children. This is a laborious approach when there are many series of maturity criteria, e.g. in the assessment of skeletal maturity. In such cases each stage is instead given a certain maturity score. The individual maturity level is calculated by averaging the maturity scores of the stages, which are then evaluated in SD-scores by comparing with the Gaussian fitted distribution of averaged maturity scores at that age. In both approaches discussed in this paper the evaluation of individual maturity is made with a relative scale of measurement. Accordingly, the rate of maturation can only be assessed indirectly.

Adolescent

Effect of maturation media on male pronucleus formation in pig oocytes matured in vitro.

The present study was carried out to examine the effect of maturation media on male pronucleus formation of pig oocyte matured and fertilized in vitro. Follicular oocytes collected from prepubertal gilts at a local slaughter house were cultured (36 h) in three different media (mTCM-199, Waymouth MB 752/l, and mTLP-PVA), fertilized in vitro, and assessed for nuclear maturation and male pronucleus formation. The addition of 10% (v/v) pig follicular fluid (pFF) to maturation media significantly increased the rate of nuclear maturation of pig oocytes (P less than 0.01), whereas the rate of nuclear maturation of pig oocytes among three different media did not differ. However, the rate of male pronucleus formation of pig oocytes was significantly higher in pig oocytes matured in Waymouth MB 752/l with or without pFF than in oocytes matured in the other two media (P less than 0.01). In experiment 2, the addition of cysteine (the same concentration as in Waymouth medium, 0.57 mM), to mTLP-PVA significantly increased the rate of male pronucleus formation of pig oocytes compared with the control (P less than 0.01). The results indicate that the composition of maturation medium affects the ability of pig oocytes to form male pronuclei following sperm penetration; media containing a high concentration of cysteine (possibly as a substrate of glutathione), such as Waymouth MB 752/l, can remarkably promote this ability.

Animals

Studies on oocyte maturation of the medaka, Oryzias latipes. VI. Relationship between the circadian cycle of oocyte maturation and activity of the pituitary gland.

The relationship between pituitary activity and oocyte maturation was examined in Oryzias latipes (medaka), which has a circadian cycle of oviposition. Throughout the circadian cycle of oviposition, females possessed a population of large oocytes more than 800 micronmeter in diameter that could mature in the presence of gonadotropin. Oocyte maturation was observed in vitro in females hypophysectomized between three and ten hours after the beginning of the light period with the number of maturing oocytes increasing as hypophysectomy was delayed. Although in vivo oocyte maturation was blocked by hypophysectomy within two hours after the beginning of the light period, it was restored by a single injection of synthetic or mammalian pituitary hormones (gonadotropic, corticotropic and thyrotropic hormones) within ten hours after hypophysectomy. Of these pituitary hormones, FSH, LH and TSH could induce in vitro maturation of isolated oocytes. Oocytes matured in vitro in the absence of exogeneous hormones if they were isolated nine or more hours after the onset of light. The present study indicates that the circadian cycle of maturation of Oryzias oocytes is controlled by the release of pituitary hormone between three and nine hours after the beginning of the light period.

Animals

Pubertal growth and maturity pattern in early and late maturers. A prospective longitudinal study of Swedish urban children.

Pubertal growth in height and the pattern of skeletal and pubertal development were studied in early and late maturers of each sex. The growth rate in height in relation to age at peak height velocity was found to be significantly greater in early maturers of both sexes from at least 7 years before to 2 years after PHV. The peak height was greater in early than in late maturers, the difference being statistically significant in boys only. The amount of growth during the pubertal growth spurt was significantly greater in the early maturers of each sex. The lag periods between the occurrence of maturity indicators of skeletal and pubertal development in relation to age at peak height velocity differed between early and late maturers. The maturity indicators occurred earlier in relation to PHV in the late maturer of each sex.

Adolescent

IL-1 beta maturation: evidence that mature cytokine formation can be induced specifically by nigericin.

Mouse peritoneal macrophages stimulated with LPS produce large amounts of pro-IL-1 beta. When these cells were pulse-labeled with [35S]methionine, however, little labeled cytokine appeared in the medium after a chase, and that which was externalized was not processed to its mature biologically active form. In an effort to promote proteolytic maturation of IL-1 beta, macrophages were treated with agents that were expected to compromise their viability. The calcium ionophore A23187 and the detergent saponin caused complete release of nonprocessed 35-kDa pro-IL-1 beta and liberation into the extracellular medium of the cytoplasmic marker enzyme LDH and the lysosomal enzyme beta-N-acetylglucosaminidase. Hypotonic lysis resulted in the release of a 20-kDa IL-1 beta species that was distinct from the 17-kDa mature species. Importantly, incubation of the murine macrophages with the potassium/proton ionophore nigericin led to a quantitative conversion of pro-IL-1 beta to a 17-kDa species. The N-terminus of this nigericin-derived product possessed the amino acid sequence expected for mature biologically active IL-1 beta. Monensin, an ionophore similar to nigericin, did not induce release or proteolysis of IL-1 beta. Complete release of mature IL-1 beta required concentrations of nigericin in excess of 2 microM and a minimum of 10 min of treatment. Mature 17-kDa IL-1 beta was observed within the nigericin-treated cells before their lysis. Nigericin's effect was not limited to mouse peritoneal macrophages, inasmuch as the ionophore also induced release and proteolytic maturation of IL-1 beta produced by LPS-stimulated human peripheral blood monocytes. Treatment of macrophages with LPS and nigericin, therefore, results in a unique series of intracellular events that promote formation of mature 17-kDa IL-1 beta.

Animals

Studies on oocyte maturation of the medaka, Oryzias latipes. V. On the structure of steroids that induce maturation in vitro.

The response of oocytes within isolated follicles (800-950 micron in diameter) to various steroids was examined with the teleost fish, Oryzias latipes. Continuous exposure of oocytes, which were removed from ovarian investments 17 hours before predicted germinal vesicle breakdown (GVBD), to C19- or C21-steroids brought about maturation in vitro but never triggered ovulation. The steroids effective in inducing maturation have in common a C=0 (or alpha-OH) group at 3C and a beta-OH group at 17C in the C19-steroids, and a C=O (or beta-OH) group at 3C and a C=O (or alpha-OH) group at 20C in the C21-steroids, in addition to an delta4- or delta5-unsaturated for 5alpha-saturated configuration. The orientation of the hydrogen at 5C seems to be critical in determining the ability of a particular steroid to stimulate oocyte maturation. Maturation of oocytes in the ovaries of hypophysectomized females was induced by administering progesterone, but the mature oocytes did not subsequently undergo ovulation. Thus the steroid hormone is capable of inducing oocyte maturation but apparently does not participate directly in the ovulation of Oryzias latipes oocytes.

Androstanes

Induction of maturation (meiosis) in small Xenopus laevis oocytes by injection of maturation promoting factor.

Treatment of small-(stage III) or medium-sized (stage IV) Xenopus laevis oocytes with progesterone, human chorionic gonadotropin, or para-hydroxymercuriphenylsulfonate does not induce maturation. Only the full-grown oocytes (stage VI) undergo maturation when treated with either one of these three substances. In contrast, injection of maturation promoting factor into oocytes of stages III-VI invariably leads to chromosome condensation and germinal vesicle breakdown. No maturation spindle is found in oocytes smaller than 0.9 nm in diameter, and the nuclear sap does not mix with the cytoplasm in the smallest (0.45-0.55 mm in diameter) oocytes. In oocytes of 0.9 mm, maturation is identical to that of full-grown oocytes, except that the maturation spindle does not reach the cortex of the oocyte. Progesterone increases protein synthesis in medium-sized (0.8 mm in diameter) oocytes without inducing meiosis. It has little or no effect on protein synthesis in smaller oocytes.

Animals

Meiotic maturation of mouse oocytes in vitro: inhibition of maturation at specific stages of nuclear progression.

In vitro studies of meiotic maturation of mouse oocytes have been carried out in the presence of several drugs. The individual steps of nuclear progression, including dissolution of the nuclear (germinal vesicle) membrane, condensation of dictyate chromatin into compact bivalents, formation of the first metaphase spindle, and extrusion of the first polar body, are each susceptible to one or more of these drugs. Germinal vesicle breakdown, the initial morphological feature characteristic of meiotic maturation, is inhibited by dibutyryl cyclic AMP. However, even in the presence of dibutyryl cyclic AMP, the nuclear membrane becomes extremely convoluted and condensation of chromatin is initiated but aborts at a stage short of compact bivalents. Germinal vesicle breakdown and chromatin condensation take place in an apparently normal manner in the presence of puromycin, Colcemid, or cytochalasin B. Nuclear progression is blocked at the circular bivalent stage when oocytes are cultured continuously in the presence of puromycin or Colcemid, whereas oocytes cultured in the presence of cytochalasin B proceed to the first meiotic metaphase, form an apparently normal spindle, and arrest. Emission of a polar body is inhibited by all of these drugs. The inhibitory effects of these drugs on meiotic maturation are reversible to varying degrees dependent upon the duration of exposure to the drug and upon the nature of the drug. These studies suggest that dissolution of the mouse oocyte's germinal vesicle and condensation of chromatin are not dependent upon concomitant protein synthesis or upon microtubules. On the other hand, the complete condensation of chromatin into compact bivalents apparently requires breakdown of the germinal vesicle. Failure of homologous chromosomes to separate after normal alignment on the meiotic spindle in the presence of cytochalasin B suggest that microfilaments may be involved in nuclear progression at this stage of maturation. Cytokinesis, in the form of polar body formation, is blocked when any one of the earlier events of maturation fails to take place.

Animals

B cell deficiency progresses with lineage maturation in nude.X-linked immunodeficient mice B cell deficiency progresses with lineage maturation.

Previously we showed that unlike normal, nude, or X-linked immune deficient (xid) mice, nude.xid mice are deficient in bone marrow pre-B cell targets for Abelson murine leukemia virus transformation. We show that nude.xid bone marrow is deficient in both CD45(B220)+ and CD45(B220)- surface (s)IgM- progenitors that give rise to B cell colonies in Whitlock-Witte cultures. CD45(B220)+ precursors had normal differentiation potential in vitro. CD45(B220)- precursors differentiated into CD45(B220)+ cells at the same rate as normal controls, but acquired sIgM at a much slower rate. These results correlated with the observation that in nude.xid mice the severity of B lineage defects correlates with maturity: a profound (ninefold) deficit of sIgM+, CD45(B220)+ mature B cells, a fivefold deficit in the sIgM-, CD45(B220)+ precursors of short term B cell colonies (colonies forming within 4-5 days in Whitlock-Witte cultures), and a moderate (twofold) decrease in the frequency of sIgM-, CD45(B220)- (less mature) precursors of long term B cell colonies (colonies forming after 14 days of Whitlock-Witte culture. Thus the combination of the nude and xid mutations produces a deficiency in early B cell progenitors and the deficiency becomes more profound with further maturation. Therefore the lack of mature B cells is the result of a cascade effect. Inasmuch as bone marrow progenitors are affected, and these are the source of the vast majority of B cells, most B cells are affected by the xid mutation and the xid defect cannot be attributed to a loss of a fetal lineage of B cells. These results suggest that xid affected cells lack the capacity to progress efficiently through differentiation in the absence of an exogenous factor(s) that is dependent on the product of a normal allele at the nude locus. This product might be supplied in vivo by a T cell or T cell-dependent source and/or epithelial elements such as bone marrow stromal cells all of which are known to be affected by the nude mutation.

Animals

Development of sexual maturity in the ciliate Euplotes crassus: sources of variation in the timing of maturity.

The life styles of ciliated protists are particularly suitable for experimental analyses of certain aspects of developmental and genetic biology. The progression from sexual immaturity to maturity to senescence represents one of the most intriguing aspects of developmental programs. The extent to which progeny clones, their subclones, and testers used in the assay result in different lengths of immaturity has been investigated in Euplotes crassus. Six subclones from each of 12 progeny clones from a cross between stocks EC1 and EC2 were tested for maturity with stocks EC3, EC4, and EC5 on every transfer. Analysis of variance was used to partition the total variation in fissions to maturity into parts due to clones, subclones, and testers and the interactions between these levels. The error, interaction of subclones and testers, corresponds to a standard deviation of only 4.1 fissions, while the within clone within tester means range from 15.2 to 46.7 fissions; all levels except testers contribute significantly to the total variation. Most of the variability is attributable to clones (66%), the next most to error (16%), the next most to interaction of clones by testers (13%), and the least to subclones (5%). An a posteriori analysis examined whether the differences among clones were due to the cytoplasm of the clone ancestor (exconjugant), its mat (mating-type) locus genotype, or the mated pair it came from. None of these characteristics was able to interpret simply the large variability among clones. These results provide evidence that the transition from immaturity to maturity is quantitative and complex rather than a jump from one well-defined state to another.

Analysis of Variance

Studies related to the head-maturation pathway of bacteriophages T4 and T2:I. morphology and kinetics of intracellular particles produced by mutants in the maturation genes.

Mutants in the genes governing the maturation of the head of bacteriophage T4 and in gene 24 were studied by electron microscopy of thin sections. We define morphologically: black particles, comprising mature, stable heads and immature, fragile heads, which break down upon lysis; grizzled particles, which apparently are partially filled or partially emptied; empty large particles without DNA or core which are all the same size as normal heads; empty small particles without DNA and without core which are of the size of the tau particle, which is the prehead of phage T4. The study of single and double mutants of the maturation genes demonstrates that the phenotypes are only different by the proportions of the different particles made except for 17- where only empty small and empty large particles accumulate. The mutants in gene 24 are epistatic on all other mutants. Mutants in gene 17 are epistatic on the remaining ones. The results are consistent with the hypothesis that the products of several of the maturation genes act on DNA to render it competent for packaging while the others act directly on the particle. By this uncoupling, bypasses and abortive pathways can result.

Coliphages

The ultrasonic changes in the maturing placenta and their relation to fetal pulmonic maturity.

A practical classification of placental maturity changes has been developed based on a review of multiple ultrasound evaluations of placental texture over a 4 year period. This classification grades placentas from 0 to 3 according to specific ultrasonic findings at the basal and chorionic plates as well as within the substance of the organ itself. The placentas of 129 patients were graded according to this system at the time of ultrasound evaluation. Eighty-six patients had placentas classified as Grade 1 or greater and all of these had lecithin-sphingomyelin (L/S) ratio determinations performed. Mature L/S ratios (2.0) were found in 68% of Grade I (21/31), 88% of Grade II (28/32), and 100% of Grade III placentas (23/23). These results suggest a correlation between maturational changes of the placenta as seen by ultrasound and fetal pulmonic maturity as indicated by L/S ratio.

Amniotic Fluid

Amniotic fluid lecithin/sphingomyelin ratio, palmitic acid, palmitic acid/stearic acid ratio, total cortisol, creatinine, and percentage of lipid-positive cells in assessment of fetal maturity and fetal pulmonary maturity: a comparison.

Lecithin/sphingomyelin (L/S) ratio, creatinine, percentage of lipid-positive cells, palmitic acid, palmitic acid/stearic acid (P/S) ratio, and total cortisol were analyzed as tests for fetal maturity and fetal pulmonary maturity in 164 samples of amniotic fluid from 121 patients. Fifty samples were taken within 72 hours of delivery. The best tests for fetal maturity (37 weeks) with differential percentages were L/S ratio, palmitic acid, and P/S ratio. In the assessment of fetal pulmonary maturity, we studied an additional 174 samples in which only L/S ratio, creatinine, and lipid-positive cells were analyzed. All tests showed a high predictive value of an immature (positive) result was much less for all six parameters; the three best tests were total cortisol (33%), lipid-positive cells (26%) and L/S ratio (14%).

Amniotic Fluid

The relationship between accelerated pulmonary maturity and accelerated neurological maturity in certain chronically stressed pregnancies.

Two studies were done. In the first, 51 infants from high-risk pregnancies were evaluated for accelerated clinical neurological maturation. Eight infants had accelerated neurological maturation 3 or more weeks in excess of gestational age. In the second study, a sample of 25 infants with documented acceleration of pulmonary surfactant (lecithin/sphingomyelin ratio greater than or equal to 2 at gestational age less than or equal to 32 weeks) all infants showed accelerated neurological maturation (range 3 to 8 weeks) with the conditions studied associated with accelerated pulmonary maturation (for instance, retroplacental bleeding, prolonged rupture of membranes, placental infarction, severe toxemia, hypertensive disease, circumvallation, and amnionitis). Chronic retroplacental bleeding was associated with the most dramatic pulmonary and neurological accelerations.

Birth Weight

Involvement of the mature domain in the in vitro maturation of Bacillus subtilis precursor 5S ribosomal RNA.

A precursor of 5S ribosomal RNA from Bacillus subtilis (p5A rRNA, 179 nucleotides in length) is cleaved by RNase M5, a specific maturation endonuclease which releases the mature 5S rRNA (m5, 116 nucleotides) and precursor fragments derived from the 5' (21 nucleotides) and 3' (42 nucleotides) termini of p5A rRNA. Previous results (Meyhack, B., et al. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 3045) led to the conclusion that recognition elements in potential RNase M5 substrates mainly reside in the mature moiety of the precursor. Limited digestion of p5A rRNA with RNase T1 permitted the isolation of a number of test substrates which contained both precursor-specific segments and were unaltered in the immediate vicinity of the cleavage sites, but which differed in that more or less extensive regions of the mature moiety of the p5A rRNA were deleted. Tests of the capacity of these partial molecules to serve as substrates for RNase M5 indicate clearly that the enzyme recognizes the overall conformation of potential substrates, neglecting only the double-helical "prokaryotic loop" (Fox, G.E., & Woese, C.R. (1975) Nature (London) 256, 505).

Bacillus subtilis

Skeletal maturity, dental maturity, and eruption in young patients with Turner syndrome.

A major problem for patients with Turner syndrome is their small body height. The rapid biotechnologic development has now made treatment with growth hormone possible at a larger scale. The aim of this investigation was to evaluate skeletal maturity, dental maturity, and eruption in a group of young patients before hormone therapy. The material comprised 33 patients aged 7-16.7 years. The skeletal maturity, as judged from hand radiographs, was on an average 2.3 years retarded (p < 0.001) and showed increasing retardation with increasing age. The dental maturity, assessed from the formation stages of the permanent teeth on panoramic radiographs, was accelerated, with a mean value of 1 year (p < 0.001). The timing of clinical eruption did not differ significantly from that of our reference material; the Turner girls were on an average 3.7 months ahead. Several patients had local eruption problems, especially in the maxillary lateral segments. It is suggested that disharmony between tooth size and arch size may contribute to this problem.

Adolescent

Progesterone-induced in vitro maturation in oocytes of Notophthalmus viridescens (Amphibia Urodela) and some observations on cytological aspects of maturation.

Maturation in vitro of oocytes of the newt, Notophthalmus viridescens, is inducible with progesterone after in vivo treatment of females with gonadotropin; few oocytes mature in vitro in the absence of such gonadotropin treatment. Chromosomes of most large oocytes of animals not receiving gonadotropin are still in the lampbrush condition; chromosomes from gonadotropin-treated animals are shorter and the lateral loops are less profuse and somewhat retracted. The chromosome condition, then, can be correlated with susceptibility to progesterone induction of maturation in vitro. As maturation progresses, the germinal vesicle moves toward the surface and decreases in size, with an apparent loss of nuclear material from the centripetal end. Although lateral loops of most chromosomes disappear during the changes in the germinal vesicle, profuse loops develop during this period at the sphere loci, which were previously devoid of loops.

Animals

[Modulation of 17-alpha-hydroxy-20 beta-dihydroprogesterone or gonadotropic extract activity on the in vitro intrafollicular maturation of rainbow trout (Salmo gairdnerii) oocytes by various steroids lacking maturing activity].

The efficiency of 17 alpha-hydroxy-20 beta-dihydroprogesterone (17 alpha-20 beta Pg) or of a Trout pituitary gonadotropic extract to induce intrafollicular maturation of trout oocytes can be modulated by some steroids which do not present any direct maturing action: gonadotropic extract efficiency is lowered by estradiol and estrone, and enhanced by testosteron. As these steroids do not present a significant effect on 17 alpha-20 beta Pg induced maturation, their site of action may be located in the follicular tissues. The corticosteroids, particularly cortisol and cortisone enhance the maturing efficiency of gonadotropin and stimulate much more strongly the efficiency of 17 alpha-20 beta Pg. This suggests a direct effect on oocyte sensitivity to 17 alpha-20 beta Pg.

20-alpha-Dihydroprogesterone