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Submicroscopic localization of glycogen in mouse blastocysts developed in vivo and in blastocysts developed in vitro from two-cell embryos.

Ultrahistochemical method according to Thiéry (1967) was used to determine the occurrence and localization of glycogen in blastocysts developed in vivo and in blastocysts developed from 2-cell embryos of the mouse for 62 to 64 h in in vitro culture. The presence of glycogen was found in blastocysts of both experimental groups. Glycogen had a monoparticulate character, i.e. the form of beta-granules, localized above all in the ground cytoplasm of cells. Their size varied from 10 to 30 nm. In the blastocysts developed in the physiological uterine environment the glycogen content was relatively low, trophoblasts cells containing regularly a higher amount of glycogen particles than embryoblast cells. In the blastocysts developed in the culture medium in the presence of currently used energy sources the distribution and content of glycogen were clearly graded according to the cell types. Compared with the in vivo-blastocysts, an abnormally high amount of glycogen was observed in the cytoplasm of trophoblast cells, a medium amount in the prospective endoderm cells and the minimum amount in the prospective ectoderm cells. The authors are of the opinion that differences in the accumulation of glycogen and its occurrence in the individual cells are in connection with their position in the blastocyst and with their relation to the surrounding microenvironment. It can be judged from the findings of glycogen deposits inside autophagic vacuoles and multivesicular bodies as well as inside extracellular located sacs that simultaneously with glycogen accumulation there also proceeds its partial degradation in lysosomal structures of blastocyst cells.

Animals

Cell-cell interactions during neural development: multiple types of lateral inhibitions involved in Drosophila eye development.

Inhibitory signals of cellular differentiation from differentiating cells play an important role in regulating the number and spatial distribution of distinctive types of cells in developing tissues. Several types of inhibitory mechanisms of cellular differentiation have been identified by making full use of the developmental genetics of Drosophila compound eyes. These inhibitory mechanisms are distinct from each other in their signal transduction cascades and/or their role in the pattern formation of the developing Drosophila eye. The following events occur: firstly a diffusible protein, Scabrous (Sca), is required to confer regular spacings of the founder cells, R8 cells, or preommatidial clusters in the developing eye disc via an unknown signal transduction cascade, secondly the Notch-signalling is at least required for the single-out of the R8 cells within the pre-ommatidial cluster possibly by preventing other cells in the equivalent groups from adapting fates as R8 cells. Notch-signalling activates a simple signal cascade mediating communication between the plasma membrane and nucleus not via protein phosphorylation. In contrast, a novel diffusible ligand, Argos, was likely to be required subsequently to the selection of R8 cells. Argos was shown to inhibit the activation of a receptor tyrosine kinase, DER, and the subsequent signal transduction in the Ras/MAPK cascade (the third inhibitory mechanism). We proposed that the role of Argos is to regulate the number of differentiated cells by controlling cellular differentiation and subsequent programmed cell death. The distinct roles of these inhibitory signals in the developing Drosophila eye are discussed in detail.

Animals

Normal and abnormal visual development in kittens: insights into the mechanisms that underlie visual perceptual development in humans.

For many years the mechanisms that underlie normal and abnormal development of visual perception in human infants have been explored in anatomical and physiological studies on two species of frontal-eyed mammals, namely, cats and, to a lesser extent, monkeys. The unstated assumption underlying the investigations on cats is that despite substantial differences in the organization of the visual pathways of cats and humans, as well as quantitative differences in their perceptual abilities, principles of development established in the former also apply to humans. This review examines the extent to which this assumption may be valid. Following a review of certain anatomical peculiarities of the cat visual system, several of the differences as well as the parallels between the perceptual abilities of cats and humans are summarized. The latter similarities, as well as the larger number of parallels between the two species that can be drawn during development, attest to the validity of the choice of the cat for study of the mechanisms that underlie human visual development.

Animals

Spatio-temporally regulated expression of receptor tyrosine kinases, mRor1, mRor2, during mouse development: implications in development and function of the nervous system.

BACKGROUND: Drosophila neurospecific receptor tyrosine kinases (RTKs), Dror and Dnrk, as well as Ror1 and Ror2 RTKs, isolated from human neuroblastoma, have been identified as a structurally related novel family of RTKs (Ror-family RTKs). Thus far, little is known about the expression and function of mammalian Ror-family RTKs. RESULTS: We have identified murine Ror-family RTKs, mRor1 and mRor2. Both mRor1 and mRor2 genes are induced upon neuronal differentiation of P19EC cells. During neuronal differentiation in vitro, the expression of mRor2 is transiently induced, although that of mRor1 increases continuously. During embryogenesis, the mRor1 gene is expressed in the developing nervous system within restricted regions and in the developing lens epithelium. The expression of mRor1 is sustained in the nervous system and is also detected in non-neuronal tissues after birth. In contrast, the expression of mRor2 is detected mainly in the developing nervous system within broader regions and declines after birth. Possible relationships of mRor1 and mRor2 genes with previously identified mutants have also been examined. CONCLUSIONS: The developmental expressions of mRor1 and mRor2, in particular in the nervous system, are differentially regulated, reflecting their expression patterns in vitro. mRor1 and mRor2 may thus play differential roles during the development of the nervous system.

Amino Acid Sequence

Inhibitor development and substitution therapy in a developing country: Turkey.

Prevalence of inhibitor in developing countries, such as Turkey, where fresh frozen plasma (FFP) is still in use due to high cost of concentrates, is unknown. To determine the frequency of inhibitors in Turkish haemophiliacs exposed to blood products, 53 haemophilia A patients (age range 1-20; median: 11 years) and 12 haemophilia B patients (age range 3-20; median: 10 years), were evaluated; 31 haemophilia A patients (23 severe) received plasma-derived concentrates and 22 patients (10 severe) only FFP. No haemophilia B patients developed inhibitor, compared with seven of 53 (13%) haemophilia A patients, all with a severe defect (7/33; 21%) and treated with concentrates (7/23; 30%), whereas severe patients treated with FFP showed a lower risk to develop inhibitors (0/10, P = 0.07). Inhibitors were detected after 8-125 exposure days (median: 52). Intermediate-purity concentrates and pasteurization seemed to be linked with a higher risk of inhibitor compared to high-purity concentrates and solvent-detergent inactivation for seven patients with inhibitor. In four of seven inhibitor patients low-dose concentrate was administered at 25 IU kg-1 twice weekly and inhibitor disappeared in 1-4 months. This regimen might be recommended for immune tolerance in developing countries for its lower cost.

Adolescent

Sucrose metabolism during cotyledon development of Vicia faba L. is controlled by the concerted action of both sucrose-phosphate synthase and sucrose synthase: expression patterns, metabolic regulation and implications for seed development.

The roles of sucrose-phosphate synthase (Sps) and sucrose synthase (Sus) in developing embryos of Vicia faba have been characterized. In the cotyledons the expression of both Sps and Sus is initiated in cells differentiating into storage tissue. This stage is characterized by a switch in the carbohydrate state from a high to a low hexoses to sucrose ratio. The carbohydrate state was found earlier to be controlled by seed coat-associated invertase. During cotyledon development the Sps-enzyme undergoes a cycle of deactivation and reactivation: the activated state is associated with the prestorage phase, desiccation and germination and the deactivated state with the storage phase. Sus activity is associated with the storage phase. Sps and Sus are differentially influenced by free sugars. Feeding hexoses to storage phase cotyledons increases levels of Sps-mRNA but not Sus-mRNA, Sps activity and Sps activation state and impairs storage functions evidenced by an increased sucrose to starch ratio and a downregulation of storage protein legumin B-mRNA. Sus enzyme activity is inhibited by free hexoses in vitro. It is proposed that the changing carbohydrate state during cotyledon development controls the ratio of Sps to Sus. Sps may have some significance for the initiation of the storage process possibly decreasing hexoses and/or increasing sucrose. The relevance of the changing carbohydrate state with respect to development and storage processes is discussed.

Base Sequence

HIV-associated immune dysfunction and delayed pubertal development in a cohort of young hemophiliacs. Hemophilia Growth and Development Study.

As part of the Hemophilia Growth and Development Study (HGDS), we investigated the relationship between HIV-associated immune dysfunction and delayed pubertal development in a cohort of 333 boys and adolescents with moderate or severe hemophilia who were between the ages of 6 and 19 years at study entry in 1989. Sixty-two percent of the cohort was infected with HIV in the late 1970s and early 1980s through exposure to contaminated clotting factor concentrates. The cohort was observed during follow-up at 6-month intervals; measurements taken at each follow-up visit included Tanner stage and CD4+ cell count. This analysis of data from the first 4 years of follow-up revealed statistically significant delays in pubertal development associated with increasing levels of immune dysfunction. Our results emphasize the importance of following pubertal development in HIV-infected adolescent boys since delays in maturation may reflect underlying disease progression.

Adolescent

Expression of two members of the Wnt family during mouse development--restricted temporal and spatial patterns in the developing neural tube.

The Wnt gene family encodes a group of cysteine-rich proteins implicated in intercellular signaling during several stages of vertebrate development. This family includes Wnt-1 and Wnt-3, both discovered as activated oncogenes in mouse mammary tumors. Here we describe the molecular cloning of an additional member of the Wnt family, called Wnt-3A, and the spatial and temporal expression pattern of this gene as well as that of its close relative Wnt-3. The putative amino acid sequences of both proteins are almost 90% identical, but in situ hybridization to mouse embryo sections showed highly restricted patterns of expression of Wnt-3 and Wnt-3A, largely in separate areas in the developing nervous system. In the spinal cord Wnt-3 was expressed at low levels in the alar laminae and in the ventral horns, whereas Wnt-3A expression was confined to the roof plate. In the developing brain Wnt-3 was expressed broadly across the dorsal portion of the neural tube with a rostral boundary of expression at the diencephalon. In contrast, Wnt-3A was expressed in a narrow region very close to the midline; expression extended into the bifurcating telencephalon, in a highly localized fashion. Both Wnt-3 and Wnt-3A were expressed in the ectoderm, and Wnt-3A was also expressed in the periumbilical mesenchyme. Characteristic expression patterns of these two closely related genes suggest that Wnt-3 and Wnt-3A play distinct roles in cell-cell signaling during morphogenesis of the developing neural tube.

Amino Acid Sequence

Regulation of GM-CSF-induced dendritic cell development by TGF-beta1 and co-developing macrophages.

Using a culture system of bone marrow progenitor cells with GM-CSF and TGF-beta1, a study was performed to analyze the effect of TGF-beta1 on the development of dendritic cells (DC) and to elucidate the regulatory role of macrophages co-developing with dendritic cells. The results demonstrate that DC generated in the presence of TGF-beta1 were immature with respect to the expression of CD86, nonspecific esterase activity and cell shape. Such inhibitory effects of TGF-beta1 were dependent on FcR+ macrophages, which were depleted by panning. TGF-beta1 did not appear to inhibit the commitment of progenitor cells to the DC lineage. In addition, TGF-beta1 also acted directly on the intermediate stage of DC to prevent their over-maturation, which results in a preferential decrease in MHC class II, but not in CD86, in the presence of TNF-alpha. FcR+ suppressive macrophages were also shown to facilitate DC maturation when stimulated via FcR-mediated signals even in the presence of TGF-beta1. These results indicate that TGF-beta1 indirectly and directly regulate the development of DC and that co-developing macrophages have a regulatory role in DC maturation.

Animals

A presentation of a conceptual framework and its use in the definition of nursing development within a number of nursing development units.

This paper provides a conceptual framework upon which action plans can be built to enable registered nurses to reflect upon their work and practice activities. This conceptual framework has been used by the author in his work with various nursing development units within Yorkshire, England, in his role as their academic nurse advisor. The framework has been useful in helping nurses not only to reflect upon current work and practice activities but also as a guideline for future development. The paper provides examples of such nursing development activities and includes a personal view of how nursing development units can act as catalysts for change.

Clinical Nursing Research

Development of enzymic zonation in liver parenchyma is related to development of acinar architecture.

The appearance of the distribution patterns of the NH3-metabolizing enzymes carbamoylphosphate synthetase, glutamate dehydrogenase, and glutamine synthetase in the developing liver of an altricial species (rat) was compared with that in the developing liver of a closely related, precocial species (spiny mouse). The comparison showed that the development of hepatic acinar architecture, rather than perinatal adaptation, is responsible for the development of periportal and pericentral compartments of gene expression. Conditions that confine the expression of specific enzymes to the pericentral compartment of the acinus originate before conditions that confine the expression of (other) specific enzymes to the periportal compartment. However, whether or not the site of gene expression is restricted to specific compartments within the liver acinus, the rate of expression of the gene involved can also be adaptively regulated. Therefore, different factors appear to control the site and the rate of gene expression within one tissue.

Aging

Early heart development in the chick embryo: effects of isotretinoin on cell proliferation, alpha-actin synthesis, and development of contractions.

Isotretinoin is a potent retinoic acid used in the treatment of skin disorders. Though very effective, it is teratogenic if administered during pregnancy, and its teratogenic effect may be related to the normal activity of retinoids as signalling molecules in the embryo. Although its exact mechanism of action is unknown, it has been suggested that it causes its characteristic pattern of defects that includes heart defects, by inhibiting the migration of neural crest cells. However, other effects on cells are known. We studied early cardiac cell proliferation using incorporation of bromodeoxyuridine (BrdU) and detection with a monoclonal anti-BrdU. Proliferation in heart tissue of whole embryo cultures was inhibited in medium with 10(-6) M isotretinoin to 62% of the control level in myocardium. We studied its effects in culture on precardiac explant development in the absence of the neural crests. Culture of precardiac mesodermal-endodermal explants revealed that development of heart vesicles from the mesoderm was little affected, but the development of heartbeat was inhibited depending on dose in the 10(-5) to 10(-7) M range. The effect on development of contractions was augmented in the presence of serum; it could be duplicated by all-trans-retinoic acid, and it was reversible. Synthesis of the alpha-actin isotype, analyzed by isoelectric focusing, was found to be inhibited or delayed. The results suggest multiple effects of retinoids on growth, morphogenesis, and differentiation of early cardiac tissue, and are discussed in relation to the potential role of retinoids in early embryogenesis.

Actins

Health, population, and economic development. International health programs have an important role in promoting economic development and population control.

Health as a basic human value is particularly important to people in the developing world. Rates of economic development lower than had been hoped for and ever more steeply rising population growth have precipitated a reaction against public health programs. Among economists, agriculturalists, and even health professionals the philosophy arose that one should "hold back" on using modern weapons against disease because they are "too effective." To satisfy the recognized popular demand, simple and relatively ineffective measures of curative medicine could be substituted. It was said that the emphasis should be, instead, on agriculture, community development, education, and industrialization and that family planning should be pushed as a separate program. Documentation presented here sharply challenges such a point of view. No segment of the total development process can be effective without the other sectors.

Demography

Effect of long-term noise exposure on the developing and developed ear in the Rat.

There have been reports that the developing ear is more sensitive than the adult ear to noise-induced hearing loss. This was investigated by testing auditory function in rats, both electrophysiologically and histologically, following exposure to broad-band noise (12 h/day for 15 days) at different stages of auditory development (neonates and adults), and also in age-matched controls. An exposure of 90 dB SPL broad-band noise caused no long-term change in auditory function in either age group. A higher exposure (102 dB SPL) caused greater long-term changes in hearing in the adult compared to the young noise-exposed rats, although histology showed greater damage to hair cells in the younger animals. Therefore, functionally, the developing ear does not seem more vulnerable than the developed ear to acoustic trauma.

Age Factors

Development and aging of cholinergic synapses. I. Endogenous levels of acetylcholine and choline in developing autonomic ganglia and iris of the chick.

Acetylcholine (ACh) and choline (Ch) levels were measured by means of a sensitive micromethod in developing irises, ciliary and sympathetic ganglia of the chick, starting at 5--7 days of incubation (DI) up to 1 year of age. The neurotransmitter is present in all three organs at relatively low levels (1--10 pmol) from 5 DI (iris) or 7 DI (ciliary and sympathetic ganglion). This is followed by a rapid and sustained 9-fold increase in all three tissues up to 14 DI. Thus, it appears that low levels of ACh may be sufficient for neurotransmission to occur in the primitive ciliary ganglion and iris. After hatching, total ACh levels continue to increase up to 1 year of age in the iris and ciliary ganglion and up to 3 months in the sympathetic ganglion. The increase may depend on either an increased functional demand for the neurotransmitter or an increased number of preganglionic terminals. In general Ch levels parallel closely the levels of ACh in each organ throughout development. It is concluded that ACh and Ch are present since the earliest detectable stages of development in the ganglia and iris, and their first increase seems to be correlated to the phase of innervation of the organs. The subsequent increase probably correlates to synaptogenesis. The close relationship between Ch and ACh levels indicates a regulatory role of Ch for ACh synthesis during neuronal development.

Acetylcholine

Evaluation of education program developments: illustration of the research and development cycle.

The Research and Development (R and D) cycle is a program development model used to translate research findings into educational programs or products ready for use in the field. The R and D cycle is illustrated by describing its application to Effective Patient Teaching, a course on teaching skills for health professionals. Evaluation is a continuing part of program development. Evaluation methods need not follow a rigid formula; methods will generally change with the stages of program development and the evaluation purposes.

Educational Measurement

Macrocyst development in Dictyostelium discoideum. I. Induction of synchronous development by giant cells and biochemical analysis.

In Dictyostelium discoideum, cytological and physiological studies on macrocyst formation revealed that this process consists of at least two steps: the production of giant cells, which are believed to be formed from the fusion of cells of two opposite mating types, and the subsequent induction of macrocyst development by the giant cells. The conditions that had been considered formerly to be required for macrocyst formation, such as darkness at the presence of two cells of complementary mating types in heterothallic strains, were actually required only for the production of the giant cells. Once giant cells are produced, the surrounding cells can aggregate and form macrocysts even in the light. Furthermore, it was demonstrated that giant cells can switch the developmental mode of the surrounding cells to macrocyst formation. That is, if a critical number of the isolated giant cells are introduced into a cell population of a single strain of NC4, which normally would produce only fruiting-bodies, macrocysts are formed instead. When in the presence of giant cells, the development of macrocysts may be initiated by starvation. Therefore, if all cells are made to starve simultaneously development begins and proceeds synchronously. Using this technique of synchronous development, the developmental kinetics of enzyme activities were assayed during macrocyst and fruiting-body formation. Considerable differences in the patterns of those enzyme activities were demonstrated between the two developmental modes of D. discoideum.

Acetylglucosaminidase