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Natural killing and antibody-dependent cellular cytotoxicity in specific-pathogen-free miniature swine and germ-free piglets. II. Ontogenic development development of NK and ADCC.

The ontogenic development of natural killing (NK) and antibody-dependent cellular cytotoxicity (ADCC) in germ-free and specific-pathogen-fee (SPF) miniature swine were compared. Activities of NK and ADCC were tested by a short-term (2.5 to 4 h) 51Cr-labelled human myeloid cell line K562 and TNP-conjugated human B-cell line SB as target cells for NK and ADCC, respectively. Animals obtained by aseptic hysterectomy 3-5 days prior to term showed ADCC activities similar to adult levels but lacked NK activity. Hysterectomy-derived piglets which were colostrum-deprived and maintained in germ-free isolators developed NK activity at 3-4 weeks of age. In comparison, naturally-farrowed, colostrum-fed piglets maintained in our SPF facility developed NK activity at 2-3 weeks of age. Thereafter, there was no significant difference in the levels of either NK or ADCC between germ-free and SPF animals. This suggests that microbial flora and environment do not affect the development of effector cells for ADCC but do play some role in the maturation of NK cells during ontogeny. The difference in ontogeny of NK an ADCC further support our previous suggestion that the effector cells for NK and ADCC in swine are distinct sub-populations.

Animals

Distribution and expression of two interactive extracellular matrix proteins, cytotactin and cytotactin-binding proteoglycan, during development of Xenopus laevis. I. Embryonic development.

An immunohistochemical study of the localization of cytotactin and cytotactin-binding (CTB) proteoglycan throughout embryonic development of the anuran Xenopus laevis reveals that both appear in a restricted pattern related to specific morphogenetic events. CTB proteoglycan expression is first detected during gastrulation at the blastopore lip. Later, it is seen in the archenteron roof around groups of cells forming the notochord, somites and neural plate. Cytotactin first appears after neurulation, and is restricted to the intersomitic regions. Both molecules appear along the migratory pathways of neural crest cells in the trunk and tail. Later, cytotactin is present at sites where neural crest cells differentiate, around the aorta and in the smooth muscle coat of the gut; CTB proteoglycan is absent from these sites. In the head, cytotactin is initially restricted to the regions between cranial somites, while CTB proteoglycan is distributed throughout the cranial mesenchyme. The expression of both molecules is later associated with key events in chondrogenesis during the development of the skull. After chondrogenesis, CTB proteoglycan is distributed throughout the cartilage matrix, while cytotactin is restricted to a thin perichondrial deposit. Both molecules are expressed in developing brain. These findings are compared to studies of the chick embryo and although distinct anatomical differences exist between frog and chick, the expression of these molecules is associated with similar developmental processes in both species. These include mesoderm segmentation, neural crest cell migration and differentiation, cartilage development, and central nervous system histogenesis.

Animals

Developing rod photoreceptors from normal and mutant Rd mouse retinas: altered fatty acid composition early in development of the mutant.

The phospholipid and fatty acid contents of developing rod photoreceptor cells were determined in dissociated photoreceptor cells obtained from normal mice and from rd mice exhibiting an inherited retinal degeneration. Photoreceptors were dissociated from retinas by mechanical agitation after mild protease treatment and characterized by light and electron microscopy. Phospholipid classes were isolated by thin-layer chromatography, and fatty acyl groups separated and quantitated by capillary gas-liquid chromatography. Developing photoreceptor cells of normal retinas accumulated all phospholipid classes, but in proportions which shifted with age. The mole % contents of phosphatidylcholine (PC) and phosphatidylinositol (PI) decreased with age, whereas phosphatidylethanolamine (PE) and phosphatidylserine (PS) increased. The content of the polyunsaturated fatty acid docosahexaenoate (22:6), expressed as nmol/microgram lipid phosphorus, increased rapidly during development, whereas arachidonate (20:4) content tended to decline. Mono-unsaturated fatty acid levels (palmitoleate, 16:1; oleate, 18:1) declined with age. Among saturated fatty acids, palmitate (16:0) decreased during normal development, whereas stearate (18:0) increased. The total mass of phospholipid/photoreceptor cell in the normal, adult mouse retina was estimated to be approximately 14 pg. The total phospholipid content and mole % distribution of individual phospholipid classes in immature rd photoreceptors were similar to values for normal cells. In contrast, significant changes in fatty acid composition were detected between immature rd cells and normal cells. Rd cells generally had higher levels of saturated (myristate, 14:0; palmitate, 16:0) and monounsaturated fatty acids (oleate, 18:1) and lower levels of polyunsaturated fatty acids (arachidonate, 20:4; docosahexaenoate, 22:6), suggesting that fatty acid metabolism is altered by expression of the rd gene and/or by the associated impairment of photoreceptor cell differentiation.

Animals

[Light microscopic studies on the development of Theileria annulata (Dschunkowsky and Luhs, 1904) in Hyalomma anatolicum excavatum (Koch, 1844). I. The development in the gut of engorged nymphs (author's transl)].

A laboratory strain of H. a. excavatum was selected on high susceptibility for T. annulata through several generations. Giemsa-stained smears and wet smears of gut and gut content were studied. After engorgement of erythrocytic stages of T. annulata by the nymphs the following development was observed: 1. Erythrocytic merozoites developed to slender, spindleshaped "microgamonts" in the gut 24 to 96 hours after repletion (p. repl.). Spherical stages with a conspicuous spike developed at the same time and earlier. The "microgamonts" then form up to 4 nuclei and several flagella-like appendices. Filiform "microgametes" obviously develop from the "microgamonts". In addition, spherical stages, i.e. "macrogametes", occur. 2. Spherical "zygotes" with a vacuole-like center appear in the epithelial cells of the gut from day 5 p. repl. These "zygotes" increase steadily in size and then stain more intensely up to day 12 p. repl. 3. From day 12 p. repl. the spherical "Zygotes" change to elongate forms by a continuing process of folding. Finally, from day 13 p. repl., they extend to clubshaped kinetes. These kinetes move actively by gliding within the gut cells and from day 17 p. repl. in the haemolymph. It could not be decided yet whether these kinetes are oo- or sporokinetes.

Animals

On the development of the cerebellum of the trout, Salmo gairdneri. V. Neuroglial cells and their development.

The neuroglia of the cerebellum of Salmo gairdneri Richardson, 1836, has been studied in mature and developing specimens with light and electron microscopy. The light microscopic observations were largely carried out on Golgi material. The cerebellum of the trout contains all of the neurologlial cell types described for the mammalian cerebellum, viz. ependymal cells, Golgi epithelial cells, velate protoplasmic astrocytes, smooth protoplasmic astrocytes and oligodendrocytes. In addition two types of glial elements, which combine characteristics of ependymal cells and of velate astrocytes, are found. These elements are designated as ependymoid astrocytes and astrocytoid ependymal cells. Smooth astrocytes and oligodendrocytes were observed only in later stages of development and possibly arise from the secondary matrix. The other glial cell types, as well as transitional forms between these types, are present in rather early stages, and show a similar ultrastructure. It is plausible that all these types develop from the glioblasts produced by the ventricular matrix layer. Many glial cells are radially oriented and keep in contact with the meningeal surface throughout development. The lattice formed by matrix cells in the earliest stages, and by glial cells and the axons of granule cells later on, plays a role in directing the migration of cells. Other functions of the glia, such as dividing the cerebellar cortex in synaptic compartments, are suggested. It may be concluded that the high degree of differentiation of the teleostean cerebellum is also reflected by the morphology of the neuroglia.

Animals

Computer-assisted analysis of the developing Purkinje neuron. I. Effects of the age of the animal at the moment of explantation on the subsequent dendritic development in organotypic cultures.

Purkinje cell dendritic arborization were studied in intracellularly horseradish peroxidase-stained mature neurons grown during 30-40 days in organotypic cerebellar cultures from kittens of various ages. The effects of the age of the animal at the moment of explantation upon the subsequent dendritic and axonal development were studied in kitten cerebella of 1, 12 and 21 postnatal days old. These effects were investigated by computer-assisted methods. Qualitative data were found to be different in these 3 in vitro models explanted at 3 different ages except for the spine development which remained the same. Quantitative data, obtained from 15 measurable parameters, showed that the degree of dendritic development was higher for the Purkinje cells explanted at the latest postnatal age: the older the animal, the larger the dendritic tree in culture. A discrimination analysis permitted a recognition of the 3 differently aged populations (94% well classified cells) based on 3 selected variables, number of primary dendritic trunks, number of rows and dendritic field areas. These results lend further support to the major role played in the final dendritic arrangement by extrinsic (discriminant factors, their pre-existence in vivo being responsible for the subsequent degree of the in vitro development of the Purkinje cell dendritic arbor.

Aging

Stimulation of ornithine decarboxylase by histamine or norepinephrine in brain regions of the developing rat: evidence for biogenic amines as trophic agents in neonatal brain development.

Ornithine decarboxylase (ODC) initiates the synthesis of polyamines which play key roles in regulation of cellular development. Intracisternal administration of histamine or norepinephrine to developing rats produced age-dependent stimulation of ODC in brain. In cerebral cortex and ponsmedulla, stimulation by norepinephrine was demonstrable at postnatal day 7 and maximum stimulation occurred at about day 9. In contrast, cerebellum showed no initial reactivity to norepinephrine but still developed a large peak of response capability by day 9. In all 3 regions, the response declined rapidly thereafter during the period of major synaptogenesis of noradrenergic pathways. With histamine, none of the regions displayed ODC reactivity at 7 days postnatally; stimulation appeared by day 9, peaked at about day 11 and then declined rapidly. Thus, the trophic effect of histamine or norepinephrine toward ODC activity is present or develops postnatally and appears to terminate with synaptogenesis and onset of neurotransmitter properties of the amines.

Age Factors

Kindling in developing animals: expression of severe seizures and enhanced development of bilateral foci.

In adult rats, alternating stimulations between two limbic sites can result in one site kindling normally, while the other is retarded in an early non-generalized kindling stage. This phenomenon has been named kindling antagonism. In this report, we present data indicating that kindling antagonism does not occur in 16-day-old rats. Instead, 16-day-old rats receiving alternating stimulations in the amygdala and hippocampus develop progressively more severe seizures. Kindling with alternate stimulations is elicited at a much faster rate at the two foci compared to kindling from a single site, either the hippocampus or amygdala. All groups develop generalized seizures including seizure stages 6 and 7, consisting of wild jumping, running with vocalizations and tonus. These seizures appear after relatively few stimulations in the pup, in comparison to the adult. The results indicate that the immature brain is less able to suppress the generalization of seizures than the adult. The age-specific enhanced development of bilateral foci may be due to underdeveloped inhibitory systems and may underlie the propensity of the immature CNS to develop multifocal seizures.

Amygdala

The postnatal development of the air-righting reaction in albino rats. Quantitative analysis of normal development and the effect of preventing neck-torso and torso-pelvis rotations.

The aim of this study was to describe the ontogenesis of the air-righting reaction (ARR) in rats. The first experiment was performed on 6 newborn albino rats of both sexes and followed the development of the ARR over postnatal days 1-21. The degree of rotation achieved after falling from different heights was quantified according to a rating scheme. It appeared that the air-righting reaction is effected by a spiral movement which spreads in a cranio-caudal direction. The reaction develops between postnatal day 8 and 18. On postnatal day 10 only a few animals are able to turn their heads, this being possible only from a falling height of 60 cm and corresponding to a falling time of 350 ms. A rapid development of the reaction was found between days 10 and 14. The second experiment on 8 rats involved the use of immobilization in order to isolate the mechanisms that trigger the ARR. The immobilization prevented neck-torso rotation, torso-pelvis rotation, and both rotations in different animals. Despite the disruption of important (afferent) feedback systems, the reaction developed within the same age period as in control rats. Thus, the Magnus "chain reflex hypothesis' as basis for the ARR is rejected in favor of a central motor program hypothesis.

Age Factors

Comparison on collagen gene expression in the developing chick embryo tendon and heart. Tissue and development time-dependent action of dexamethasone.

Glucocorticoids modulate various cellular functions such as proliferation, energy metabolism and the synthesis of proteins. In the present study, the response of collagen genes to dexamethasone in different stages of chick embryo development was studied in tendon and heart using Northern blot analysis and specific cDNA probes. The changes in collagen gene expression were compared to alterations in two reference mRNAs: actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The levels of specific mRNAs measured per ribosomal RNA in tendon and heart varied markedly during normal development. In tendon the relative levels of alpha 1(I), alpha 2(I) and alpha 1(III) collagen mRNAs were highest between days 14-16 when also the synthesis of matrix proteins is most active. In heart the levels of these mRNAs peaked at day 12. In addition, qualitative differences were observed in the expression of actin genes between tendon and heart. Dexamethasone in high dose decreased collagen mRNA levels in tendons, while in heart a stimulatory effect was noted. Dexamethasone also decreased GAPDH mRNA levels in tendons. The alterations in gene expression after dexamethasone treatment in tendon and heart did not correlate with the level of specific glucocorticoid receptors, which varied markedly during the development of chick embryos. The cDNA for pro alpha 1(I) collagen hybridized to two transcripts corresponding to 6.2 and 5.1 kb in tendon and heart. During normal development of chick embryos the ratio of 6.2/5.1 kb mRNAs decreased markedly in heart, but no such change was observed in tendons. Dexamethasone, however, decreased the ratio of 6.2/5.1 kb transcripts in tendons. There was a significant correlation between the ratio 6.2/5.1 kb transcripts and total alpha 1(I) mRNA both in tendon and heart, suggesting that the 6.2 kb transcript may be associated with the rate of synthesis of type I collagen.

Actins

Middle-ear development. V: Development of umbo sensitivity in the gerbil.

PURPOSE: The development of the umbo response in the gerbil was studied in order to further elucidate the contribution of the middle ear to the development of auditory function. MATERIALS AND METHODS: Laser interferometry was used to study the development of umbo velocity in Mongolian gerbils between 10 days after birth and maturity. RESULTS: Before 15 days after birth, immaturities in the middle ear prevented any reliable measures of middle-ear motion. However, between 15 and 20 days after birth, a 10 dB improvement in umbo velocity was noted in the low-frequency (0.5 to 2.0 kHz) region of the umbo response. This improvement in sensitivity was correlated to an increased admittance due to an expanding bulla volume. Interestingly, umbo velocity remained relatively constant in the mid- and high-frequency regions of the response curve between 15 and 42 days after birth. The umbo response in the adult gerbil was decidedly different when compared with the response at 42 days after birth. CONCLUSION: We speculate that a decrease in bulla volume along with increased ossicular mass contributed to the changes in the adult umbo response. When the maturation of the umbo response was compared with more central ontogenetic measures, it became apparent that structures more central to the middle ear continued to develop well past the time the middle ear was structurally and functionally mature.

Acoustic Stimulation

Exposure of postnatal rats to glucocorticoids suppresses the development of choline acetyltransferase-immunoreactive neurons: role of adrenal steroids in the development of forebrain cholinergic neurons.

Rat forebrain cholinergic neurons undergo dynamic developmental changes, showing a continuous increase in choline acetyl-transferase (ChAT) activity, during the early postnatal period. In adult rats, increases in circulating glucocorticoids result in decreases in activity of forebrain neuronal ChAT, thus raising the possibility that postnatal development of forebrain cholinergic neurons results from low levels of these hormones. In the rat, the first 2 weeks postnatally are characterized by very low levels of adrenal steroids. To understand the role of endogenous glucocorticoids in the development of forebrain cholinergic neurons, we studied the changes in ChAT immunoreactivity in forebrain cholinergic neurons of postnatal rats which had received daily subcutaneous injection of the synthetic glucocorticoid dexamethasone for 8 days. Immunohistochemical analysis of the rat pup forebrain revealed nearly complete obliteration of ChAT-immunoreactive neurons in the caudate-putamen, especially in the dorsolateral region of the rostral part. At the same stage, treatment with dexamethasone induced significant decreases in both number and length of dendritic branches of ChAT-immunoreactive neurons in the substantia innominata and the diagonal band. Despite the marked alterations in the caudate-putamen and diagonal band, the ChAT-immunoreactive neurons in other forebrain structures such as globus pallidus and medial septal nucleus showed little change. In the caudate-putamen, Nissl staining and specific labeling for nuclear DNA fragmentation exhibited no increase in number of dying cells following dexamethasone treatment, therefore indicating that the loss of ChAT immunoreactivity is not due to glucocorticoid-induced cholinergic cell death. These observations demonstrated that the development of cholinergic neurons in rat pups was inhibited by prolonged glucocorticoid exposure, suggesting that low levels of adrenal steroids may promote the postnatal development of these neurons.

Adrenal Cortex Hormones

Oxidative influence on development and differentiation: an overview of a free radical theory of development.

Metabolic gradients exist in developing organisms and are believed to influence development. It has been postulated that the effects of these gradients on development result from differential oxygen supplies to tissues. Oxygen has been found to influence the course of development. Cells and tissues in various stages of differentiation exhibit discrete changes in their antioxidant defenses and in parameters of oxidation. Metabolically generated oxidants have been implicated as one factor that directs the initiation of certain developmental events. Also implicated as factors that modulate developmental processes are the cellular distribution of ions and the cytoskeleton both of which can be influenced by oxidants. The interaction of oxidants with ion balance and cytoskeleton is discussed.

Animals

Effect of development of antibodies to HLA and cytomegalovirus mismatch on lung transplantation survival and development of bronchiolitis obliterans syndrome.

OBJECTIVE: A retrospective analysis was performed to examine the role of HLA antibodies and cytomegalovirus mismatch on the development of bronchiolitis obliterans syndrome and survival after lung transplantation. METHODS: Of 339 consecutive lung transplantations performed over a 102-month interval, 301 patients survived at least 3 months. There was a minimum follow-up period of 13 months. Bronchiolitis obliterans syndrome was defined as a decline in forced expiratory volume in 1 second less than 80% of posttransplantation baseline and/or histologic presence of obliterative bronchiolitis and was defined as occurring "early" if documented within 3 years of transplantation. Variables analyzed included preoperative donor and recipient cytomegalovirus status and the development of antibodies to human leukocyte antigens after transplantation. Microcytotoxicity was used to determine the presence of antibodies to human leukocyte antigens. Variables were subjected to Kaplan-Meier analysis to determine their impact on freedom from bronchiolitis obliterans syndrome and survival. RESULTS: The development of antibodies to human leukocyte antigens after transplantation correlated significantly with bronchiolitis obliterans syndrome (P = .02). The development of antibodies to human leukocyte antigens did not affect survival (P = .33) unless they were detected within 2 years of transplantation (P = .04). There was greater frequency of early bronchiolitis obliterans syndrome in cytomegalovirus seronegative patients who received allografts from seropositive donors compared with all other combinations (P = .02). There was also a trend toward worse survival of cytomegalovirus seronegative patients who received allografts from seropositive donors (P = .13). CONCLUSION: These data suggest that bronchiolitis obliterans syndrome is the result of an immune-mediated process in which HLA antibodies and cytomegalovirus may play a significant role.

Actuarial Analysis

The case for early identification of hearing loss in children. Auditory system development, experimental auditory deprivation, and development of speech perception and hearing.

Human infants spend the first year of life learning about their environment through experience. Although it is not visible to observers, infants with hearing are learning to process speech and understand language and are quite linguistically sophisticated by 1 year of age. At this same time, the neurons in the auditory brain stem are maturing, and billions of major neural connections are being formed. During this time, the auditory brain stem and thalamus are just beginning to connect to the auditory cortex. When sensory input to the auditory nervous system is interrupted, especially during early development, the morphology and functional properties of neurons in the central auditory system can break down. In some instances, these deleterious effects of lack of sound input can be ameliorated by reintroduction of stimulation, but critical periods may exist for intervention. Hearing loss in newborn infants can go undetected until as late as 2 years of age without specialized testing. When hearing loss is detected in the newborn period, infants can benefit from amplification (hearing aids) and intervention to facilitate speech and language development. All evidence regarding neural development supports such early intervention for maximum development of communication ability and hearing in infants.

Acoustic Stimulation

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