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PubMedDate not supplied

Establishment of an immortalized Copenhagen rat prostate epithelial cell line.

An immortal cell line of Copenhagen rat prostate epithelium was established after transfection of primaly cultured cells with SV4O DNA and designated EPYP-1. The established cells have continued to proliferate more than 70 population doublings, have not undergone senescence, and stain positively for SV40 T-antigen in their nuclei by immunohistochemistry. They grow in a monolayer with an epithelial morphology and exhibit positive staining for cytokeratin. EPYP-1 cells exhibited positive staining with anti-prostatic acid phosphatase antibody and expressed Integrin a6,B1 on their plasma membrane. They did not demonstrate Dil-Ac-LDL uptake as an endothelial marker. Both testosterone and dihydrotestosterone stimulated the growth of EPYP-1. Genetically EPYP-1 was aneuploid, however no tumors formed after subcutaneous and intra-prostatic injection of the cells in Copenhagen rats. This immortalized cell line of Copenhagen rat prostate epithelial origin may be suitable for studying early events in the conversion of cells to tumorigenicity.

Acid Phosphatase↗
PubMed1997

Calpain activation contributes to dendritic remodeling after brief excitotoxic injury in vitro.

The calcium-dependent protease calpain may contribute to neuronal death in acute neurological insults and may be activated very early in the neuronal injury cascade. We assessed the role of calpain in a model of rapid, reversible dendritic injury in murine cortical cultures. Brief sublethal NMDA exposure (10-30 microM for 10 min) resulted in focal swellings, or varicosities, along the length of neuronal dendrites as visualized with the lipophilic membrane tracer Dil or with immunostaining using antibodies to the somatodendritic protein MAP2. These varicosities appeared within minutes of NMDA exposure and recovered spontaneously within 2 hr after NMDA removal. Addition of the calpain inhibitors MDL28,170, calpain inhibitors I and II, and leupeptin (all 1-100 microM) had little effect on the development of NMDA-induced dendrite injury. However, the resolution of varicosities was substantially delayed by addition of calpain inhibitors after sublethal excitotoxic exposure. Using Western blots and immunocytochemistry, we observed reactivity for a calpain-specific spectrin proteolytic fragment during the period of recovery from dendritic swelling, but not during its formation. Spectrin breakdown product immunoreactivity could be blocked by the calpain inhibitor MDL28,170 and appeared in neuronal cell bodies and neurites in a time course that paralleled dendritic recovery. These observations suggest that calcium-dependent proteolysis contributes to recovery of dendritic structure after NMDA exposure. Calpain activation is not necessarily detrimental and may play a role in dendritic remodeling after neuronal injury.

Animals↗
PubMed1997

Growth cone form is behavior-specific and, consequently, position-specific along the retinal axon pathway.

Video time-lapse microscopy has made it possible to document growth cone motility during axon navigation in the intact brain. This approach prompted us to reanalyze the hypothesis, originally derived from observations of fixed tissue, that growth cone form is position-specific. The behaviors of Dil-labeled retinal axon growth cones were tracked from retina through the optic tract in mouse brain at embryonic day (E) 15-17, and these behaviors were matched with different growth cone forms. Patterns of behavior were then analyzed in the different locales from the retina through the optic tract. Throughout the pathway, episodes of advance were punctuated by pauses in extension. Irrespective of locale, elongated streamlined growth cones mediated advance and complex forms developed during pauses. The rate of advance and the duration of pauses were surprisingly similar in different parts of the pathway. In contrast, the duration of periods of advance was more brief in the chiasm compared to those in the optic nerve and tract. Consequently, in the chiasm, growth cones spent relatively more time pausing and less time advancing than in the optic nerve or tract. Thus, because growth cone form is behavior-specific and certain behaviors predominate in particular loci, growth cone form appears to be position-specific in static preparations, due to the fraction of time spent in a given state in different locales.

Animals↗
PubMed1997

Epileptic activity prevents synapse formation of hippocampal mossy fibers via L-type calcium channel activation in vitro.

Hippocampal slice from early postnatal rat was used to elucidate the influence of epileptic activity elicited by picrotoxin on synapse formation of mossy fibers. Neurite reelongation and synaptogenesis of mossy fibers transected at 8 days in vitro were confirmed by staining with Dil, a fluorescent membrane dye used as a neuronal tracer, and by recording field excitatory postsynaptic potentials (fEPSP) in the CA3 region evoked by stimulation of the dentate gyrus. Picrotoxin (50 microM), which evoked spontaneous epileptiform firing in the CA3 region that was occluded by tetrodotoxin (1 microM), hindered development of fEPSP amplitude after a lesion of mossy fibers. Furthermore, observations using a Timm method, a histochemical technique that preferentially labels synaptic terminals of mossy fibers, revealed that picrotoxin prevented synaptogenesis in the CA3 region. This inhibitory effect of picrotoxin was completely abolished by tetrodotoxin or nicardipine (10 microM), a L-type calcium channel blocker, but not by 2-amino-5-phosphonopentanoic acid (50 microM), a N-methyl-D-aspartate receptor antagonist, suggesting that influx of calcium ion via L-type calcium channels during epileptic bursts mediated the disturbance of appropriate synapse formation of mossy fibers.

Animals↗
PubMed1997

[Antithrombotic property of new growing endothelial cells on the prosthetic vascular grafts--experimental study of portal vein replacement].

On experimental portal vein replacement of mongrel dogs with high porosity EPTFE grafts (fibril length: 60 microns) wrapped in omental pedicle flap, it has been reported that healing process is promoted. But it is not clear on the antithrombotic properties of the newly growing endothelial cells on prosthetic vascular grafts. PGI2 and NO are known as antithrombotic factors that restrain the agglutination of platelets. We evaluated PGI2 and NO production from the newly growing endothelial cells on prosthetic vascular grafts. High porosity EPTFE grafts were implanted at the portal veins of mongrel dogs. After 3 months we removed the grafts and tried to procure the new growing endothelial cells by trypsinization and culture them. We confirmed by using Dil-Ac-LDL that endothelial-like cells on prosthetic vascular grafts were as same as the endothelial cells. As control cells, we used the endothelial cells of canine portal vein and infra vena cava. The basal PGI2 production from each cells and the thrombin-stimulated production of PGI2 were measured by radioimmuno assay. Thrombin concentration was established at 2 unit/ml. On the basal PGI2 production there was no significant difference between the grafts and the portal veins, and between the grafts and the infra vena cava. The PGI2 production of the grafts increased significantly by stimulating with thrombin. On the thrombin-stimulated production, there was no significant difference between the grafts and the portal veins, and between the grafts and the infra vena cava. NO production of the grafts was measured by the griess reaction. On NO production there was no significant difference between the grafts and the portal veins. But the grafts produced significantly more NO than the infra vena cava (P < 0.05). This study suggests that the newly growing endothelial cells on the prosthetic vascular grafts may have the antithrombotic properties equal to the endothelial cells on the portal vein and the infra vena cava.

Animals↗
PubMed1997

Development of the anterior commissure in the opossum: midline extracellular space and glia coincide with early axon decussation.

While the anterior commissure has been shown to be an important route of information transfer in the forebrain, relatively little is known about its anatomical development. Glial substrates and extracellular spaces have been associated with the maturation of other large-fiber tracts, such as the corpus callosum and retinofugal pathway. The present study examined early stages in the maturation of the commissure in the gray short-tailed opossum, Monodelphis domestica. Monodelphis offspring are born after a short 14-day gestation, and, unlike in rats and mice, the anterior commissure develops entirely during the postnatal period. A number of techniques were employed: the carbocyanine dye Dil was used to label early axons in the region, semithin plastic sections were used to examine the extracellular environment of the developing commissure, and immunocytochemistry for glial fibrillary acidic protein (GFAP) was used to characterize glial components. Results suggest that the first commissural fibers that cross the midline pass through a region of large extracellular spaces and may use GFAP-immunoreactive cells and processes as guides during their midline decussation.

Animals↗
PubMed1997

Local origin of cells in FGF-4 - induced outgrowth of amputated chick wing bud stumps.

Urodele amphibians are the only vertebrates that can regenerate amputated limbs, even as adults. However, we have previously shown that amputated chick wing bud stumps can be induced to ((regenerate)) and to form a complete set of correctly-patterned skeletal elements, following implantation of beads soaked in fibroblast growth factor-4 (FGF-4). We have now performed Dil injection experiments to determine which cells contribute to FGF-4-induced chick wing bud ((regenerates)). We show that the FGF-4-induced outgrowth of the regenerating wing bud stump is comprised of mesenchyme cells that originate from a region within 200 microm of the FGF-4 bead, and that cells proximal to the bead move distally.

Animals↗
PubMed1998

A mouse mandibular culture model permits the study of neural crest cell migration and tooth development.

A major issue in developmental biology is to determine how time and position-restricted instructions are signaled and received during morphogenesis of different phenotypes, of which tooth, Meckel's cartilage and tongue formation are classical examples. It is now evident that a hierarchy of growth factors and their downstream transcription factors regulate the timing, sequence and position of cells and tissues in forming different phenotypes during embryogenesis. Here we report the development of an early mandibular organ culture model. Explants of E8 and E9 first branchial arch were cultured and produced mandibular processes with cap stage tooth formation, Meckel's cartilage and tongue development. In tandem, vital dye (Dil) labeling studies confirmed that rhombomeres 1-4 give rise to craneal neural crest (CNC) cells which emigrate from the neural fold to the forming maxillary and mandibular arches. Furthermore, we have tested the feasibility of investigating the regulation of different phenotypes within the first branchial arch by a transcription factor using this early mandibular organ culture model. Lymphoid enhancing factor 1 (Lef1), a transcription factor, has been implicated to regulate tooth formation in vivo. We have analyzed the expression of Lef1 and studied the biological effects of Lef1 on E8 embryonic mouse first branchial arch explants in organ culture. Collectively, these results demonstrate that first branchial arch explant model is suitable for studies of rhombencephalic crest cell fate during mandibular morphogenesis and can be used as a model with direct access to investigate the molecular mechanism in regulating first branchial arch morphogenesis.

Animals↗
PubMed1998

[A case of male who was taken systemic lupus erythematosus with chilblain lupus].

A 38-year-old male was admitted to our hospital because of high grade fever, erythema of face and extremities and oral ulcer. The laboratory examination showed leukopenia, high titer (320 dil.) of antinuclear antibody, a positive reaction of anti-Sm antibody. Especially histopathology of hand erythema showed hydrophilic degeneration consist with chilblain lupus. Because his symptoms were consistent with the criteria of American College of Rheumatology (1982), he was diagnosed as systemic lupus erythematosus (SLE). After administration of 60 mg prednisolone daily, the symptoms gradually improved. Any recrudescence of serological abnormalities or clinical symptoms was not observed until now, although chilblain lupus still persisted. Originally SLE is frequent in young female and often attended with renal disease, central nerve disease and serositis. We reported a case of middle-aged male who developed SLE with chilblain lupus which was rare skin lesion of SLE without severe organic lesion and discussed the relation among chilblain lupus, discoid lupus erythematosus (DLE) and SLE.

Adult↗
PubMed2003

NEDD8 protein is involved in ubiquitinated inclusion bodies.

Proteolysis by the ubiquitin-proteasome system is considered to play a pathological role in several degenerative diseases that involve ubiquitinated inclusion bodies. In recent years, several ubiquitin-like proteins have been isolated, but it is uncertain whether their roles are associated with protein degradation through the ubiquitin-proteasome system. NEDD8 (neural precursor cell-expressed and developmentally down-regulated gene), which consists of 81 amino acid residues, possesses the highest sequence similarity to ubiquitin. Recent studies have indicated that NEDD8 is covalently ligated to cullin family proteins, which are components of certain ubiquitin E3 ligases, by a pathway analogous to that of ubiquitin. Thus, by focusing on the structural and functional association between NEDD8 and ubiquitin, it would be of interest to know whether the NEDD8 system is involved in pathological disorders of the ubiquitin-proteasome system. This study has examined the immunohistochemical distribution of NEDD8 protein by using a highly purified antibody in normal tissues and in tissues known to contain ubiquitinated inclusions. NEDD8 protein expression was widely observed in most types of tissues. Furthermore, accumulation of the NEDD8 protein was commonly observed in ubiquitinated inclusion bodies, including Lewy bodies in Parkinson's disease, Mallory bodies in alcoholic liver disease, and Rosenthal fibres in astrocytoma. Two of ten cases of neurofibrillary tangles and senile plaques from patients with Alzheimer's disease showed intense staining for NEDD8 as well as for ubiquitin. These findings suggest the possibility that the NEDD8 system is involved in the metabolism of these inclusion bodies via the ubiquitin-proteasome system.

Alzheimer Disease↗
PubMed2026

A Review on Heat Stress in Broiler Chickens: Mechanisms, Effects and Mitigation Strategies.

BACKGROUND: Heat stress (HS) is a major environmental challenge for broilers, particularly under rising global temperatures and high humidity. Broiler chickens are highly susceptible because of their rapid growth rate, high metabolic heat production, limited thermoregulatory capacity and genetic selection for fast growth. OBJECTIVE: This review aims to synthesise current evidence, evaluate the effectiveness of existing mitigation strategies, identify key knowledge gaps and provide future research directions to improve broiler resilience, welfare and productivity under increasingly HS conditions. METHODS: This review synthesised evidence published between 2010 and 2025 on the physiological, metabolic, intestinal, immunological and productive consequences of HS and evaluated mitigation strategies. RESULTS: The reviewed studies demonstrate that HS reduces feed intake by approximately 10%-30%, suppresses body weight gain and feed efficiency and increases mortality, with severity depending on temperature, humidity and broiler genotype. HS disrupts carbohydrate, protein and lipid metabolism; induces acid-base imbalance and oxidative stress; compromises intestinal barrier integrity; alters gut microbiota; suppresses immune function; and reduces meat quality. Nutritional interventions, including dietary electrolyte balance, antioxidants, vitamins, selenium, zinc, phytogenic compounds, probiotics, betaine and optimised feeding strategies, environmental management and genetic approaches, including naked-neck and frizzle genes, can partially alleviate these adverse effects. However, inconsistencies among studies persist because of differences in broiler strains, environmental conditions, dietary formulations and experimental protocols. CONCLUSION: HS substantially compromises broiler health, welfare, productivity and meat quality. Nutritional, environmental and genetic approaches can partially mitigate its adverse effects; however, further research is needed to improve broiler resilience under increasingly HS conditions.

Animals↗
PubMed1979

A double-blind pediatric evaluation of levamisole in the prevention of recurrent upper respiratory tract infections.

Levamisole was tested double-blind in 106 children with recurrent upper respiratory tract infections. They received either levamisole (n = 53) or placebo (n = 53) 0.5 ml/kg bodyweight b.i.d. for two consecutive days each week for six months. A control examination was performed every two months. Both groups were compared by means of the Fisher-test and the Mann-Whitney U-test (two-tailed probability each). Improvement was observed more frequently in the levamisole group with regard to the number of episodes of infection, and the total duration and severity of the infections. No side-effects, except for some stomach complaints in one levamisole patient, were reported.

Child↗