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At least 901 records · Page 50Linked to original sources

PubMed1985

Development of a mouse antiperoxidase secreting hybridoma for use in the production of a mouse PAP complex for immunocytochemistry and as a parent cell line in the development of hybrid hybridomas.

Mouse antibodies are increasingly used as primary antibodies for immunocytochemistry as more mouse monoclonal antibodies are being produced. The localisation of these antibodies by the PAP technique requires mouse antiperoxidase antibody. A monoclonal antiperoxidase would obviate the limitations of production of a polyclonal mouse antiperoxidase. This paper describes the development of a mouse hybridoma producing such an antibody (MAP A6-2) and the use of this antibody to localise a number of mouse primary antibodies by the PAP technique for both light and electron microscopy. The antibodies localised include monoclonal antienkephalin and antityrosine hydroxylase. MAP A6-2 had a higher affinity in immuno-diffusion experiments and gives slightly better staining with an horse radish peroxidase of a different type from that used for immunisation. Staining was optimum with horse radish peroxidase type X whereas horse radish peroxidase type VI was used for immunisation. Also described is the production of a HAT sensitive variant cell line allowing the possibility of using this hybridoma as a parent cell line for the production of hybrid hybridomas secreting bi-specific antibodies.

Animals↗

PubMed1990

Development under the influence of cocaine. II. Comparison of the effects of maternal cocaine and associated undernutrition on brain myelin development in the offspring.

Pregnant and lactating Long-Evans rats were treated daily with oral cocaine at a dosage rate of 60 mg/kg/day, which is the highest dosage tolerated during chronic treatment. Brain myelin concentrations were compared in the offspring during early myelination (day 15) and peak myelination (day 20). Body and brain weights in the offspring of cocaine-treated and pair-fed dams were transiently (but insignificantly) retarded, and by day 20 brain weights of the normal controls, cocaine-exposed, and pair-fed pups were the same (1.36 +/- 0.01-1.35 +/- 0.01 g). Similarly, specific myelin yields in the offspring of pair-fed dams were retarded by no more than 1-2% of the myelin concentrations obtained in normal controls, which is consistent with studies showing that such minimal growth retardation would not be expected to produce hypomyelination. In contrast, myelin concentrations from the brains of cocaine-exposed pups were reduced about 10%. In addition, cross-fostering indicated that the fetal period of cocaine exposure presents a greater risk to postnatal myelination than exposure during the suckling period, which is the reverse of the vulnerability resultant from undernourishment. Thus, the hypomyelination observed in the offspring of cocaine-treated dams indicates a toxic effect of cocaine acting directly on the developing offspring.

Aging↗

PubMed1991

Investigation of some antimicrobial procedures on the in vitro development of early murine embryos aimed toward developing methods for the disinfection of mammalian embryos prior to transfer.

Eight-cell, zona pellucida-intact mouse embryos were exposed to the following substances or procedures that have been reported to have germicidal effects to determine if the embryos would survive and develop under in vitro conditions: the photosensitive substances hematoporphyrin, hematoporphyrin derivative, 8-methoxypsoralen, 4,5',8-trimethylpsoralen, and thiopyronine; the enzymes lipase (0.5%), phospholipase C (2 U/ml), chymotrypsin (0.5%), and trypsin (0.5%); pH 5.0; and helium/neon laser light, visible light, ultraviolet A light, and ultraviolet C light. Under the conditions used, embryos were not adversely affected by hematoporphyrin and/or helium/neon laser light; methoxypsoralen and/or ultraviolet A light; lipase; trypsin; pH 5.0 for 20 min; and visible light. Variable results were obtained from hematoporphyrin derivative with laser light. Thiopyronine, trimethylpsoralen in combination with ultraviolet A light, and ultraviolet C light killed embryos, and chymotrypsin and phospholipase C were harmful at 10- and 15-min exposure times, respectively.

Animals↗

PubMed1987

Adipose tissue development: the role of precursor cells and adipogenic factors. Part I: Adipose tissue development and the role of precursor cells.

Obesity is regarded as a heterogeneous syndrome, which may appear in different forms. Various causes have been found to contribute to its pathogenesis. During recent years investigations of adipose tissue cellularity and its dynamic changes have gained growing interest. An important progress was the discovery of adipose tissue precursor cells. These cells have not yet been precisely identified by morphological and biochemical methods in intact tissue. However, due to methodological developments such precursor cells can be cultured both as primary cultures and as established cell lines. These culture systems have proven to be valuable models for the study of the processes involved in the formation of new fat cells.

Adipose Tissue↗

PubMed1987

Histamine response in developing chick oesophagus. A pharmacological analysis of the response to histamine in the oesophagus of developing chick.

Developmental changes in response to histamine were investigated in the oesophagus isolated from the chick between 15 days of incubation and 15 days after hatching. The contraction could be already caused by histamine (20 microM) in the chick oesophagus from 15 days of incubation. The pD2 values for histamine and acetylcholine did not change during the period between 17 days of incubation and 5 days after hatching. The response to histamine (20 microM) was extremely resistant to tetrodotoxin (0.78 microM), procaine (0.2 mM) and atropine (1 microM) during the period between 17 days of incubation and 20 days of incubation. The tetrodotoxin-resistant response was replaced by a tetrodotoxin-sensitive response after hatching. On the other hand, the neuronal response induced by transmural electrical stimulation (20 Hz) or dimethylphenylpiperazinium (20 microM) was uniformly sensitive to tetrodotoxin throughout the periods. Mepyramine (2 microM), but not metiamide (20 microM), inhibited the histamine-induced responses in every age tested. These results suggest that the myogenic receptivity of histamine transiently increases during the terminal period of embryonic development and declines after hatching.

Acetylcholine↗

PubMed1988

Noradrenergic innervation of developing rat and spiny mouse liver. Its relation to the development of the liver architecture and enzymic zonation.

The development of noradrenergic innervation of rat liver was studied with a polyclonal antiserum against noradrenaline. Nerves are first seen in the larger portal vessels at day 1 after birth and reach their final distribution at 5 days after birth i.e. at the same time as the establishment of the acinar architecture and the heterogeneous distribution of NH3-metabolizing enzymes. The latter distribution of nerves is already seen at birth in the liver of the closely related but precocial spiny mouse. This shows that the onset of extrinsic sympathetic innervation is regulated by the developmental stage of the animal rather than by adaptation to extrauterine life. Chemical sympathectomy at birth with 6-hydroxydopamine did not eliminate the developmental appearance of heterogeneous distributions of NH3-metabolizing enzymes.

Ammonia↗

PubMed2001

Development of muscle pathology in canine X-linked muscular dystrophy. II. Quantitative characterization of histopathological progression during postnatal skeletal muscle development.

We have characterized the time course of muscle pathology development during the postnatal maturation of quadriceps and tibialis anterior muscle in dystrophic golden retriever dogs. We determined the percentages of degenerating, regenerating, calcium-positive, hypercontracted, albumin-positive, and C3 complement fraction-positive muscle fibers and the extent of connective tissue proliferation in animals from neonate to adult. Necrotic fibers increased from days 2 to 30, decreased at 60 days (to 0.8%) and increased in older animals to a stable level of around 2%. Hypercontracted fibers peaked at 15 days (19.1%) and declined to 3.7% in adults. Regenerating fibers were numerous at 15 and 30 days (10%), declined at 60 days to 4.7% and declined further in adults. Calcium- and albumin-positive fibers peaked at 30 days (6.5% and 13.8%, respectively) and then declined to around 3% and 5%, respectively, in older dogs. In dystrophic dogs, the extent of fibrosis was significantly greater on 15 days than in controls, but did not then increase with age. In carriers, calcium- and albumin-positive fibers always expressed dystrophin abnormally. Muscle damage occurs before completion of muscle maturation in dystrophic dogs. While necrosis and hypercontraction remain stable in adults, fiber regeneration declines to very low levels. In contrast to Duchenne muscular dystrophy, muscle fibrosis in the muscle studied does not increase with age.

Aging↗

PubMed1997

The multidrug-resistance P-glycoprotein (Pgp, MDR1) is an early marker of blood-brain barrier development in the microvessels of the developing human brain.

The multidrug-resistance P-glycoprotein (Pgp) was initially identified as an energy-dependent proton pump, which transports a variety of non-related compounds out of chemotherapy-resistant cancer cells. Molecular biological investigations using knockout mice for the mouse homologue of the human Pgp showed that these mice partially lack a functioning blood-brain barrier, indicating that Pgp has an important role in the blood-brain barrier as its normal function. The presence of Pgp expression in formalin-fixed and wax-processed tissue sections can be assessed using the monoclonal antibody, JSB-1. Since no data on the developmental expression of Pgp are available, we stained a developmental series of human brain sections with JSB-1. Our results indicate that Pgp expression in endothelia of brain microvessels occurs regularly in embryos of about 30-mm crown-rump length (CRL). Strong reactivity is seen in blood vessels of fetuses from 123-mm CRL. There is also reactivity in pial blood vessels but not in choroid plexus blood vessels known to be without a blood-brain barrier. Pgp expression is therefore an early marker of the blood-brain barrier in the developing human brain.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2005

Development and reliability of a standard rating system for outcome measurement of foot and ankle disorders I: development of standard rating system.

BACKGROUND: The aim of this study was to report the five scales comprising the rating system that the Japanese Society for Surgery of the Foot (JSSF) devised (JSSF standard rating system) and the newly offered interpretations and criteria for determinations of each assessment item. METHODS: We produced the new scales for the JSSF standard system by modifying the clinical rating systems established by the American Orthopaedic Foot and Ankle Society (AOFAS scales) and the Japanese Orthopaedic Association's foot rating scale (JOA scale). We also provided interpretations of each assessment item and the criteria of determinations in the new standard system. RESULTS: We improved the ambiguous expressions and content in the conventional standard rating systems so they would be easily understood by Japanese people. The result was five scales in total. Four were designed for use specifically for ankle-hindfoot, midfoot, hallux metatarsophalangeal-interphalangeal, and lesser metatarsophalangeal-ineterphalangeal sites; and the fifth was for the foot and ankle with rheumatoid arthritis. Furthermore, we described interpretations and criteria for determinations with regard to evaluation items in each scale. CONCLUSIONS: Conventionally, the AOFAS scales or the JOA scale have been separately applied depending on the sites or disorders concerned, but it was often difficult to decide on scores during practical evaluations because of differing expressions in different languages and also because of ambiguity in the interpretation of each evaluation item and in scoring standards as well. JSSF improved these scales and added definite interpretations of evaluation items as well as criteria for the rating (to be reported here in part I). Because these steps were expected to improve the reliability of outcomes assessed by each scale, we examined the reliability in scores of the newly developed scales, which are reported in part II (in this issue).

Ankle↗

PubMed2006

Novel semi-automated methodology for developing highly predictive QSAR models: application for development of QSAR models for insect repellent amides.

Conventional 3D-QSAR models are built using global minimum conformations or quantum-mechanics based geometry-optimized conformations as bioactive conformers. QSAR models developed using the global minima as bioactive conformers, employing the GFA, PLS and G/PLS methodologies, gave good non-validated r(2) (0.898, 0.868 and 0.922) and performed well on an internal validation test with leave-one-out correlation q(2) (LOO) (0.902, 0.726 and 0.924), leave-10%-out correlation q(2) (L10O) (0.874, 0.728 and 0.883) and leave-20%-out q(2) (L20O) (0.811, 0.716 and 0.907). However, they showed poor predictive ability on an external data set with best predictive r(2) (Pred-r(2)) of 0.349, 0.139 and 0.204 respectively. A novel methodology to mine bioactive conformers, from clusters of conformations with good 3D-spatial representation around pharmacophoric moiety, furnishes highly predictive 3D-QSAR models. The best QSAR model (model A) showed r(2) of 0.989, q(2) (LOO) of 0.989, q(2) (L10O) of 0.980, q(2) (L20O) of 0.963 and Pred-r(2) on eight test compounds of 0.845. The methodology is based on mimicking the multi-way Partial Least Squares (PLS) technique by performing several automated sequential PLS analyses. The poses/shapes of the mined bioactive conformers provide valuable insight into the mechanism of action of the insect repellents. All of the repetitive tasks were automated using Tcl-based Cerius2 scripts.

Algorithms↗

PubMed1983

Glucose transport in developing Ehrlich ascites tumor cells: parallel changes in the rate of glucose uptake and cytochalasin B binding activity during tumor development and methotrexate treatment.

The ability of Ehrlich ascites tumor cells to take up glucose increased progressively during the course of tumor development. Simultaneously as the rate of uptake rose, the density of a class of glucose-reversible binding sites for cytochalasin B on the cell surface also increased. In its stereospecificity requirement toward competing sugars and in its sensitivity to phloretin and diethylstilbestrol, this class of binding sites resembled the putative glucose carriers identified in various other cell systems and may represent the glucose transporter in Ehrlich ascites cells. Work with methotrexate (MTX) substantiated this view. Methotrexate arrested tumor growth, inhibited glucose uptake, and reduced the number of cytochalasin B binding sites. In both MTX-treated and untreated cells, the magnitude of changes in number of cytochalasin B binding sites closely paralleled and sufficiently accounted for the magnitude of changes in glucose uptake. Qualitative changes in the turnover and affinity for substrate of the putative glucose carrier need not be invoked.

Animals↗

PubMed1979

The development of estrogen receptor systems in the rat brain: perinatal development.

The estrogen receptor system of the rat brain is not detected at 15 and 19 days of gestation, even though estrogen receptors can be measured in the maternal hypothalamus. Estrogen receptors are first detected around the 21st day of gestation after which they increase rapidly in the perinatal period and reach a plateau around postnatal day 6. Estrogen receptors are found predominantly in the limbic brain (hypothalamus, preoptic area, amygdala) and in the cerebral cortex, and they are present in very low levels in a sample consisting of the midbrain, brain stem and cerebellum. Development of cortical estrogen receptors is retarded by a few days compared to those in the limbic brain, but then is extremely rapid so that cortical receptor levels are equal to those in the limbic block on or about postnatal day 6. As early as estrogen receptors can be detected in the fetus (day 21 of gestation), they are occupied by endogenous estradiol in male but not in female fetuses. This is measured by the occurrence of cell nuclear estrogen receptor complexes which can be detected by an exchange assay. The presence of such complexes in male limbic brain tissue is consistent with the presence of testicular androgen in the circulation and with the existence in the brain of aromatizing enzymes capable of converting testosterone to estradiol. The absence of such complexes from the female limbic brain is consistent with the protective role of alpha-fetoprotein in scavenging circulating maternal estradiol and keeping it from reaching intracellular receptor sites. The results are discussed with respect to the timing of the critical period for brain sexual differentiation and related maturational events.

Adrenalectomy↗

PubMed1979

The development of estrogen receptor systems in the rat brain and pituitary: postnatal development.

The postnatal development of the estrogen receptor system of the rat brain is described by means of data from in vitro cytosol binding assays and from in vivo cell nuclear experiments using 3H-labeled estrogens. Cytosol and nuclear measures give a similar picture of the changes in estrogen receptor levels from day 2--3 up to day 25 of postnatal life. Pituitary receptor levels reach a peak at day 10 and then decline slightly. A similar, though less pronounced, peak was seen in hypothalamus between days 8 and 15. In the amygdala, the concentration of receptors remains relatively constant apart from a slight increase around day 10. In the preoptic area, receptor levels increase throughout the entire period studied. In the cortex receptor levels increase between days 3 and 10 and then decline precipitously and remain low from day 15 onwards. In the midbrain and brain stem receptor levels remain low throughout the entire period. Occupation of estrogen receptors by estradiol in cell nuclei was investigated by means of an exchange assay. No occupation was seen in either cortex or limbic structures (hypothalamus, amygdala, preoptic area) of female pups on postnatal days 2--3; low level occupancy amounting to around 5% of capacity was seen in limbic structures but not in cortex of females on postnatal days 10--11 and 25--26. The possibility is discussed that this estrogen arises from testosterone via aromatization. The apparent absence of receptor occupation by endogenous circulating estradiol is discussed in relation to the presence and progressive disappearance of alpha-fetoprotein in the neonatal period. Data is presented showing that injected estrogens have a longer half-life in newborn rats than in 3-week-old animals and that estrogens such as diethylstilbestrol and moxestrol, which do not bind strongly to alpha-fetoprotein, gain access to brain estrogen receptors at lower doses compared to estradiol-17 beta.

Aging↗

PubMed1994

Targeted disruption of the BDNF gene perturbs brain and sensory neuron development but not motor neuron development.

Brain-derived neurotrophic factor (BDNF), a neurotrophin, enhances the survival and differentiation of several classes of neurons in vitro. To determine its essential functions, we have mutated the BDNF gene. Most homozygote mutants die within 2 days after birth, but a fraction live for 2-4 weeks. These develop symptoms of nervous system dysfunction, including ataxia. The BDNF mutant homozygotes have substantially reduced numbers of cranial and spinal sensory neurons. Although their central nervous systems show no gross structural abnormalities, expression of neuropeptide Y and calcium-binding proteins is altered in many neurons, suggesting they do not function normally. In contrast with mice lacking the BDNF receptor TrkB, motor neurons appear normal in the BDNF mutant.

Animals↗

PubMed1980

Influence of unilateral castration and increased plane of nutrition on sexual development of Holstein bulls. II. Histologic development of the testes.

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).

Journal Article↗

PubMed1988

In vitro culture and maintenance of trophoblastic vesicles: Observations on the development of contractile structures with concurrent vessel development.

The use of trophoblastic vesicles as a model for the study of maternal recognition of pregnancy, embryo development and cell to cell interaction has increased in recent years. In this report, we describe the long-term culture of trophoblastic vesicles derived from enzymatically dispersed porcine blastocysts. Day 12 to 14 porcine embryos were enzymatically dispersed to form single cell suspensions, then allowed to plate in 25 cm(3) tissue-culture flasks. Vesicles formed 3 to 14 d following plating and ranged in size from 0.1 to 4.0mm in diameter. Vesicles larger than 1.5 mm at formation were able to float free in the medium for up to 65 days, at which time vesicles were histologically sectioned. Contractile elements formed in association with dense cell areas, and were found with tubular vascular vessels. Movement of cells that resembled eosinophilic cells was observed in the vessels and pulsed in synchrony with contractions of the primitive heart. Histological examination gave evidence of primitive blood cells and cardiac tissue, and no evidence of neural tissue was found, indicating the probability that the contractions were induced by permeable membrane depolarization. It is hypothesized from the observations reported in this paper that the cell differentiation required to form the contractile region and the vessel are under the control of the dense cell area. This indicates that trophoblastic vesicles fromed by the enzymatic dispersal of porcine blastocysts is a possible model for the study of induced cell differentiation and formation of the cardiac organ system in vitro.

Journal Article↗

PubMed1983

The correlation between the development of sympathetic innervation and the development of medial hypertrophy in jejunal arteries in normotensive and spontaneously hypertensive rats.

The catecholaminergic innervation of jejunal arteries was examined during development in normotensive and hypertensive rats. It was determined that a hyperinnervation of jejunal vessels in the hypertensive rats is present from 2 weeks of age onwards, before significant elevation of blood pressure and medial hypertrophy of jejunal arteries occurs. It is concluded that this hyperinnervation may be causally related to the medial hypertrophy in arteries of hypertensive animals.

Age Factors↗

PubMed1983

Transitory macrophages in the white matter of the developing visual cortex. II. Development and relations with axonal pathways.

Clusters of 'gitter cells' develop in the white matter of the occipital cortex of the cat at the end of the first postnatal week. These clusters, and others already present at birth, disappear by the end of the first postnatal month. The life span of the clusters in the occipital white matter corresponds to the period when transitory callosal axons are eliminated. The clusters have close contact with callosal axons and can be labeled by HRP injected in the contralateral hemisphere and transported through the corpus callosum. One of the clusters clearly forms in a part of the white matter crossed by transitory callosal axons. The 'gitter cells' might be involved in the elimination of these axons. Consistent with this hypothesis, ultrastructural observations show groups of axons completely surrounded by 'gitter cell' cytoplasm as if they were being phagocytosed.

Aging↗