TNF-alpha induces apoptosis in rat fetal brown adipocytes in primary culture

The balding (bal) mutation of the mouse is an autosomal recessive mutation that causes alopecia and immunologic anomalies. A new allele was identified by allelism testing after using an interspecific backcross to localize the mutation to the centromeric end of mouse chromosome 18. We investigated the skin and hair histologic lesions of two alleles (bal(J) and bal(Pas)) at this locus and analyzed the expression of several keratinocyte markers and the production of autoantibodies by immunofluorescence on frozen skin sections. The lesions observed included separation of the inner and outer root sheath in anagen follicles resulting in the hair fiber being very easily plucked from the follicle. Vesicles on the ventral tongue, mucocutaneous junction of the eyelid, foot pads, and rarely in skin were also evident. Separation occurred between the basal and suprabasilar cells forming an empty cleft, resembling that observed in human pemphigus vulgaris. Immunofluorescence studies did not reveal the presence of tissue-bound or circulating autoantibodies. Expression of keratinocyte markers in hair follicles was normal. Keratin 6-positive cells were found on either side of the follicular separation suggesting a molecular defect in adhesion molecules between the inner layer of the outer root sheath cells to layers on either sides. This hypothesis has been confirmed by another group who demonstrated that the bal(J) mutation is due to the insertion of a thymidine in the desmoglein 3 gene, resulting in a premature stop codon.     

DDT induces apoptosis in rat thymocytes

We have isolated an Arabidopsis BBM II isomerase cDNA from an Arabidopsis cDNA library, by means of functional complementation of the E. coli hisA mutant strain HfrG6. The isolated cDNA encodes a polypeptide of 304 amino acids with a calculated molecular weight of 33,363. Sequence comparison with the HIS6 proteins of yeasts revealed that Arabidopsis BBM II isomerase contains an N-terminal extension of approximately 40 amino acids that shows the general properties of chloroplast transit peptides. This finding is consistent with the localization of other histidine biosynthetic enzymes, such as imidazoleglycerolphosphate dehydratase and histidinol dehydrogenase, in the chloroplasts in higher plants. The primary structure of the mature protein was 50% and 42% identical, respectively, to the HIS6 proteins of Schizosaccharomyces pombe and Saccharomyces cerevisiae, respectively, while no prominent sequence similarity to the bacterial BBM II isomerase was found. That the isolated Arabidopsis cDNA actually encodes a functionally active BBM II isomerase activity was confirmed in an in vitro enzyme assay using a crude extract prepared from strain HfrG6 transformed with the Arabidopsis BBM II isomerase cDNA.     

Adriamycin induces apoptosis in rat thymocytes

Apoptosis is a controlled form of cell death accompanied by distinct morphological and biochemical changes. In this study the nature of cytotoxicity induced by adriamycin (ADM) in rat thymocytes was evaluated. Morphological and biochemical changes characteristic of apoptosis were found to precede adriamycin-induced cell death. Our findings demonstrate the involvement of c-Myc, c-Jun, antioxidant enzymes CuZn superoxide dismutase and catalase, and perhaps poly ADP ribosylation in ADM-induced cell death.     

Cyclosporin A induces apoptosis in rat hepatocytes in culture

Salmonella typhimurium phage type (PT) or definitive type (DT) 104 is a virulent pathogen for humans and animals, particularly cattle. It has been isolated increasingly from humans and animals in the United Kingdom and several other European countries and, more recently, in the United States and Canada. Humans may acquire the infection from foods of animal origin contaminated with the infective organism. Farm families are particularly at risk of acquiring the infection by contact with infected animals or by drinking unpasteurized milk. The symptoms in cattle are watery to bloody diarrhea, a drop in milk production, pyrexia, anorexia, dehydration and depression. Infection may result in septicemic salmonellosis and, upon necropsy, a fibrinonecrotic enterocolitis may be observed. The infection occurs more commonly in the calving season than at other times. Feedlot cattle and pigs may also be affected. Prolonged carriage and shedding of the pathogen may occur. Symptoms in humans consist of diarrhea, fever, headache, nausea, abdominal pain, vomiting, and, less frequently, blood in the stool. Salmonella typhimurium DT104 strains are commonly resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline.     

Macrophage-derived nitric oxide induces apoptosis of rat hepatoma cells in vivo

Although nitric oxide (NO) has been postulated to play important roles in host defense mechanisms against tumor cells, a direct evidence supporting this hypothesis is lacking. To obtain molecular insights into the antitumor action of NO, its metabolism and effect on ascites hepatoma (AH-130) cells were investigated in tumor-bearing rats. Kinetic analysis revealed that substantial amounts of nitrite and nitrate, metabolites of NO, appeared in plasma and ascites of AH-130-inoculated rats. Western blot analysis revealed that a large number of macrophages that expressed inducible type of NO synthase (iNOS) appeared in cancerous ascites, particularly during 1 to 2 weeks after inoculation of AH-130 cells. When NO generation by peritoneal macrophages increased, a significant fraction of AH-130 in ascites fluid underwent apoptosis as judged from the fragmentation of their nuclear DNA. Kinetic analysis revealed that NO strongly inhibited mitochondrial electron transport and changed calcium status in AH-130 cells, particularly under low oxygen tensions such as in cancerous ascites. Intraperitoneal injection of NO donor strongly enhanced DNA fragmentation of AH-130 cells. Antimycin A, a specific inhibitor for mitochondrial electron transport, also induced DNA fragmentation of AH-130 cells by a mechanism that was inhibited by adding ascorbate and tetramethyl-p-phenylene diamine (TMPD) as electron donors. These results indicate that NO derived from peritoneal macrophages inhibits mitochondrial electron transport and disturbs calcium homeostasis in ascites hepatoma AH-130 cells, thereby inducing their apoptosis in vivo.     

Development of normal mice from metaphase I oocytes fertilized with primary spermatocytes

Primary spermatocytes are the male germ cells before meiosis I. To examine whether these 4n diploid cells are genetically competent to fertilize oocytes and support full embryo development, we introduced the nuclei of pachytene/diplotene spermatocytes into oocytes that were arrested in prophase I (germinal vesicle stage), metaphase I, or metaphase II (Met II). Both the paternal and maternal chromosomes then were allowed to undergo meiosis synchronously until Met II. In the first and second groups, the paternal and maternal chromosomes had intermingled to form a large Met II plate, which was then transferred into a fresh enucleated Met II oocyte. In the third group, the paternal Met II chromosomes were obtained by transferring spermatocyte nuclei into Met II oocytes twice. After activation of the Met II oocytes that were produced, those microfertilized at metaphase I showed the best developmental ability in vitro, and three of these embryos developed into full-term offspring after embryo transfer. Two pups (one male and one female) were proven to be fertile. This finding provides direct evidence that the nuclei of male germ cells acquire the ability to fertilize oocytes before the first meiotic division.     

Involvement of c-myc in the resistance of non-obese diabetic mice to glucocorticoid-induced apoptosis

We conducted a multicentered, retrospective review of clozapine's (CZP) effects on a range of psychiatric, sleep, cognitive, motor, and sensory disorders in Parkinson's disease (PD). Therapeutic outcomes and adverse events were compared with varying prescribing practices at participating sites. The medical records of 172 consecutive PD patients treated with CZP at four movement disorder clinics were reviewed. Low-dose CZP improved psychiatric symptoms of psychosis, anxiety, depression, hypersexuality, sleep disturbance, and akathisia. Tremor, torticollis, limb dystonia, and pain showed modest rates of improvement. Twenty-three percent of patients withdrew as a result of adverse events or treatment failure. Inpatient CZP initiation did not improve therapeutic efficacy, or reduce adverse events or the withdrawal rate. Low-dose CZP in the outpatient setting is generally an effective and well-tolerated treatment for many of the psychiatric, sleep, motor, and sensory disturbances common to late-stage PD.     

Termination of lactation induces apoptosis and alters the expression of the Bcl-2 family members in the rat anterior pituitary

After lactation, there is a massive loss of pituitary lactotrophs. The aim of this study was to investigate the events and mechanisms involved in the remodeling of the anterior pituitary after termination of lactation. Animals were analyzed at day 7 of lactation and at 4 and 7 days after pup removal. Field-inversion gel electrophoresis was used to detect high-molecular-weight DNA fragments, a characteristic feature of apoptosis. Bax and Bcl-2, proteins involved in regulation of cell survival, were studied by immunohistochemistry and Western blot analysis. In addition, the expression of Bax, Bcl-2, Bcl-X(L), and p53 messenger RNA (mRNA) was analyzed by in situ hybridization. Four days after termination of lactation, a peak in high-molecular-weight DNA fragments and an increase in Bax, along with a decrease in Bcl-2 proteins, were observed, in comparison with lactation and with 7 days post lactation. The mRNAs for Bax and p53 also were increased, whereas no changes were detected in Bcl-2 and Bcl-X(L) mRNA levels. In summary, these findings indicate that the cell remodeling of the anterior pituitary, after the termination of lactation, occurs through the process of apoptosis and involves changes in Bax, Bcl-2, and p53.     

Human immunodeficiency virus type 1 Tat protein induces death by apoptosis in primary human neuron cultures

Hamartomas are rare benign tumors appearing very often without any symptoms. In case of diagnostic detection the operative exploration is indicated for a correct histological diagnosis and resection. In this report we describe a very rare case of mesenterial hamartoma. During the late postoperative course a refilling chylothorax and chylous ascites occurred. With total parenteral nutrition, excluding short- and long-chain fatty acids, chylous leakage was successfully treated. The chylous exudation was stopped totally; only intraabdominally did minimal ascites persist. The therapy was continued by a oral Ceres diet nutrition. The conservative therapy involving total parenteral nutrition permitted us to avoid the peritoneovenous Le Veen shunt and the associated complications.     

Mannitol at clinical concentrations activates multiple signaling pathways and induces apoptosis in endothelial cells

To examine the characteristics of Ha?ssaguerre's slow potential (SP) specific to effective catheter ablation of the slow pathway in AV nodal reentrant tachycardia, the properties of SP and its recording site were analyzed in 52 patients who underwent successful SP-guided ablation. The properties of SP included the ratio of the amplitude of SP to that of atrial potential (A)(SP/A), the SP duration, the interval between His-bundle potential (HP) and SP (HP-SP), the interval between A and SP (A-SP), the interval between SP and ventricular potential (V) (SP-V), and the ratio of A-SP to the interval between A and the V (A-SP/A-V). The SP recording site was determined by the ratio of the amplitude of A to that of V (A/V) and by the relative position of the ablation catheter on X ray (right anterior oblique projection), expressed as the ratio of the distance between the coronary sinus ostium and SP site to that between the coronary sinus ostium and HP recording site (relative SP position). Twenty-eight slow pathways were ablated with a single energy application, while the other 24 required applications > or = 2. In all successful applications, SP/A, SP duration, HP-SP, A-SP, SP-V, A-SP/A-V, A/V, and relative SP position were 51% +/- 25%, 28 +/- 5 ms, -11 +/- 9 ms, 57 +/- 25 ms, 68 +/- 13 ms, 46% +/- 9%, 15% +/- 13%, and 51% +/- 13%, respectively. A significant correlation was observed between the relative SP position and A-SP, and between the relative SP position and A-SP/A-V (r = 0.60 and 0.37, respectively), while it was not between the relative SP position and HP-SP, nor between the relative SP position and SP-V. When the characteristics of SP were comparatively analyzed between the effective and ineffective applications in 24 patients in whom applications > or = 2 were required, there was no difference observed in HP-SP, A-SP, SP-V, A-SP/A-V, and A/V. However, SP/A, SP duration, and the relative SP position in the effective applications were all greater than those in the ineffective ones (56% +/- 20% vs 35% +/- 18%, P < 0.001; 29 +/- 4 vs 26 +/- 5 ms, P < 0.01; and 52% +/- 15% vs 33% +/- 11%, P < 0.001, respectively). These results indicate that SP with an amplitude over a half of A amplitude and recorded at the mid-septum of the tricuspid annulus can be a marker for successful slow pathway ablation. Although the local atrial electrogram appears late as the SP recording site shifts to the lower position, the timing of SP relative to HP and V remained unchanged, suggesting that SP is independent of the local atrial activation.     

Disruption of epithelial cell-matrix interactions induces apoptosis

Cell-matrix interactions have major effects upon phenotypic features such as gene regulation, cytoskeletal structure, differentiation, and aspects of cell growth control. Programmed cell death (apoptosis) is crucial for maintaining appropriate cell number and tissue organization. It was therefore of interest to determine whether cell-matrix interactions affect apoptosis. The present report demonstrates that apoptosis was induced by disruption of the interactions between normal epithelial cells and extracellular matrix. We have termed this phenomenon "anoikis." Overexpression of bcl-2 protected cells against anoikis. Cellular sensitivity to anoikis was apparently regulated: (a) anoikis did not occur in normal fibroblasts; (b) it was abrogated in epithelial cells by transformation with v-Ha-ras, v-src, or treatment with phorbol ester; (c) sensitivity to anoikis was conferred upon HT1080 cells or v-Ha-ras-transformed MDCK cells by reverse-transformation with adenovirus E1a; (d) anoikis in MDCK cells was alleviated by the motility factor, scatter factor. The results suggest that the circumvention of anoikis accompanies the acquisition of anchorage independence or cell motility.     

TRAIL induces apoptosis and activation of NFkappaB

TRAIL (TNF-related-apoptosis-inducing-ligand) was found as a new member of the TNF family which mediates cell death in a wide variety of malignant cell lines and primary tumor cells. TRAIL induces two different signals, cell death mediated by caspases and gene induction mediated by NFkappaB. Inhibition of TRAIL-induced activation of NFkappaB augments apoptosis induction by TRAIL and attenuates apoptosis resistance.     

Immunocytology of chiasmata and chromosomal disjunction at mouse meiosis

The presented paper is focused on substantial features of neuropsychological examinations. The author emphasizes the application of different tests focused on the relationship between the brain and behaviour, but in particular the neuropsychological battery. He describes the Halstead--Reitan battery and possibilities of its application in children. He emphasizes the importance of neuropsychological rehabilitation and differentiates it from mere recording of the process of spontaneous recovery. He characterizes the main neuropsychological factors which affect recovery after traumatic brain damage. He mentions some of his own examples where he used these factors in clinical practice. The author describes also his own experience with computer rehabilitation using the SERMION--Train the Brain programme.     

Overexpression of the human type-1 insulin-like growth factor receptor in rat L6 myoblasts induces ligand-dependent cell proliferation and inhibition of differentiation

The human type-1 insulin-like growth factor receptor was overexpressed in rat L6 skeletal myoblasts using a retroviral expression vector. At low concentrations of insulin-like growth factor-I (1-10 ng/ml), dose-dependent stimulation of myoblast differentiation was similar in control L6 myoblasts and L6 myoblasts which overexpressed the type-1 insulin-like growth factor receptor. At high concentrations of insulin-like growth factor-I (50-100 ng/ml), L6 myoblasts which overexpressed the receptor did not undergo terminal differentiation and proliferated to form multilayers, while differentiation of control myoblasts was further stimulated. These findings indicate that overexpression of the type-1 IGF receptor can stimulate proliferation and inhibit differentiation in skeletal myoblasts in a ligand-dependent manner.     

In vitro analysis of ovarian differentiation and the initiation of meiosis in the rat

Intact, cranial, and caudal portions of fetal rat gonads on days 13, 14, 15, 16 and 17 of gestation were cultured singly on Nucleopore filters in Eagle's MEM and McCoy's 5A media (both supplemented with fetal calf serum and glutamine) to evaluate the role of the mesonephros in mammalian gonadogenesis. After 2--7 days in culture, in vitro and in vivo control tissues were examined by light and electron microscopy to determine the extent of morphological differentiation and gonium maturation. The onset of morphological differentiation and meiotic activity, as determined by synaptonemal complex formation, was found to be independent of the mesonephric epithelium. Ovaries and testes were also cultured as heterologous pairs on opposite sides of collagen-coated Nucleopore filters for 4--6 days in McCoy's 5A media. In the absence of transfilter cellular migration and establishment of cell-to-cell contact, the in vivo pattern of sexual dimorphic onset of meiosis was observed. This finding lends support to the concept that local somatic-germ cell associations as opposed to extragonadal factors, are capable of providing the necessary environment for regulation of meiosis in the fetal rat.