Exogenous histone H1 injection into mitotic cells disrupts synchronous progression of mitotic events by delaying chromosome decondensation

1994 ◽  
Vol 107 (3) ◽  
pp. 693-701 ◽  
Author(s):  
Y. Matsuoka ◽  
S. Takechi ◽  
T. Nakayama ◽  
Y. Yoneda
Keyword(s):  
Mitotic Spindle ◽  
Protein Transport ◽  
Type A ◽  
Histone H1 ◽  
Type B ◽  
Lamin B ◽  
Calf Thymus ◽  
Exogenous Histone ◽  
Chromosome Decondensation

At the end of open mitosis, chromosome decondensation, nuclear envelope re-formation and reassembly of interphase microtubules following mitotic spindle dissociation occur coordinately. To determine whether these events progress only synchronously in vivo, we delayed chromosome decondensation by injecting of exogenous proteins into the mitotic rat kangaroo kidney epithelium (PtK2) cells. When histone H1 purified from calf thymus was injected at prometaphase, chromosome condensation was prolonged for several hours, and sister chromatid separation and cytokinesis did not occur. However, interphase microtubules reassembled and lamin B-positive structures re-formed around the condensed chromosomes. Exactly the same results were obtained on injection of bacterially expressed H1. Kinetic experiments showed that there were two types of lamin B-positive structures. One type (type A) was stained uniformly with anti-lamin B antibodies. The other (type B) showed peripheral lamin B staining; that is, the normal interphase staining pattern, and was found to be competent for nuclear protein transport. As the chromosomes decondensed, the amount of type A decreased and that of type B increased. However, even cells containing highly condensed chromosomes had both type A and type B. From these results, we conclude that the re-formation of microtubules and reassembly of a nuclear transport-competent envelope do not depend on chromosome decondensation.

scholarly journals Two Alternatively Spliced Transcripts from the Drosophila period Gene Rescue Rhythms Having Different Molecular and Behavioral Characteristics

1998 ◽  
Vol 18 (11) ◽  
pp. 6505-6514 ◽  
Author(s):  
Yuzhong Cheng ◽  
Barbara Gvakharia ◽  
Paul E. Hardin
Keyword(s):  
Type A ◽  
Behavioral Characteristics ◽  
Cdna Clones ◽  
Type B ◽  
Differential Splicing ◽  
Rnase Protection ◽  
Period Gene ◽  
Alternatively Spliced ◽  
Transgenic Flies

ABSTRACT The period (per) and timeless(tim) genes encode key components of the circadian oscillator in Drosophila melanogaster. The pergene is thought to encode three transcripts via differential splicing (types A, B, and C) that give rise to three proteins. Since the threeper mRNA types were based on the analysis of cDNA clones, we tested whether these mRNA types were present in vivo by RNase protection assays and reverse transcriptase-mediated PCR. The results show that per generates two transcript types that differ only by the presence (type A) or absence (type B′) of an alternative intron in the 3′ untranslated region. Transgenic flies containing transgenes that produce only type B′ transcripts (perB′ ), type A transcripts (perA ), or both transcripts (perG ) rescue locomotor activity rhythms with average periods of 24.7, 25.4, and 24.4 h, respectively. Although no appreciable differences in type A and type B′ mRNA cycling were observed, a slower accumulation of PER in flies making only type A transcripts suggests that the intron affects the translation ofper mRNA.

scholarly journals The binding of adenosine(5′)tetraphospho(5′)adenosine to calf thymus histones measured by non-equilibrium dialysis

1987 ◽  
Vol 246 (3) ◽  
pp. 681-686 ◽  
Author(s):  
G Just ◽  
E Holler
Keyword(s):  
Ionic Strength ◽  
Dissociation Constants ◽  
Equilibrium Dialysis ◽  
Physiological Saline ◽  
Histone H1 ◽  
The Other ◽  
Competition Assay ◽  
Calf Thymus ◽  
Non Equilibrium

Binding of adenosine(5′)tetraphospho(5′)adenosine (Ap4A) to histones of calf thymus was investigated by non-equilibrium dialysis. Histone H1 interacts with the dinucleotide via two strong sites and competes with Mg2+ ions. Intrinsic dissociation constants were 1.6 +/- 0.1 microM and 11 +/- 1 microM for zero and 0.4 mm-Mg2+ concentration respectively. Binding of poly(dT) and of other nucleotides to histone H1 was measured in an [3H]Ap4A-competition assay. The tendency to form complexes among nucleotides was highest for bisnucleoside tetraphosphates and decreased in the order poly(dT) greater than or equal to Ap4A approximately Gp4G greater than Ap4 much greater than Ap3A approximately Ap5A greater than or equal to ATP, GTP and dTTP. The co-ordination complex derived from Ap4A and cis-diammine-dichloroplatinum(II) was not reactive. The other histones of calf thymus also bound Ap4A with affinities decreasing in the order H4 approximately H3 greater than H1 greater than H2b greater than H2a. Ap4A stimulated the exchange of histone H1 between nucleosomes, but this effect was referred to ionic strength. It did not bind to assembled nucleosomes. Binding of Ap4A to histone H1 was decreased by salt (NaCl). At physiological saline concentration the value of the dissociation constant is commensurable with the value of the Ap4A concentration in the nucleus and thus indicative of complex-formation in vivo.

Downmodulation of IL-8 receptors, type A and type B, on human lung neutrophils in vivo

1997 ◽  
Vol 273 (3) ◽  
pp. L618-L625 ◽  
Author(s):  
K. Soejima ◽  
S. Fujishima ◽  
H. Nakamura ◽  
Y. Waki ◽  
M. Nakamura ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Type A ◽  
Type B ◽  
Low Level ◽  
Blood Neutrophils ◽  
Vitro Analysis ◽  
High Level ◽  
In Vitro Analysis

We examined the expression of interleukin (IL)-8 receptors (Rs), type A (IL-8-RA) and type B (IL-8-RB), on peripheral blood and bronchoalveolar lavage (BAL) fluid neutrophils; we also examined IL-8 and other chemoattractants in the epithelial lining fluid (ELF) of patients with chronic lower respiratory tract infection (CLI) to elucidate the in vivo regulation of IL-8Rs. Neutrophils were stained with monoclonal antibodies specific for IL-8-RA and IL-8-RB. We detected higher levels of IL-8 (81.6 +/- 25.4 ng/ml, mean +/- SE), leukotriene (LT) B4, and IL-1 beta in the ELF of the CLI patients than in their serum (P < 0.05). The expression of IL-8Rs on BAL neutrophils was significantly lower than that on peripheral blood neutrophils (P < 0.01 for both). In vitro analysis showed that low-level IL-8 (50 ng/ml) alone did not affect IL-8R expression but that it was downregulated by high-level IL-8 (500 ng/ml) alone and by low-level IL-8 in combination with LTB4 or IL-1 beta. Staurosporine reduced the downmodulation by low-level IL-8 plus LTB4 or IL-1 beta but not by high-level IL-8 alone. We speculate that pulmonary IL-8-RA and IL-8-RB may have been downmodulated by the combined effect of local chemoattractants through, in part, a protein kinase C-dependent mechanism.

Viropexis and Phagocytosis of Type B (Mouse Mammary Tumor) Virus Particles in vivo

1973 ◽  
Vol 31 ◽  
pp. 580-581
Author(s):  
Gabriel Seman ◽  
Leon Dmochowski
Keyword(s):  
Tumor Cells ◽  
Type A ◽  
Reticulum Cell ◽  
Type B ◽  
Cytoplasmic Matrix ◽  
Virus Particles ◽  
Tumor Virus ◽  
Type C

Electron microscope examination of spleen tumors of RIII/Dm female mice with reticulum cell sarcoma revealed the presence of extracellular and budding type C, type B, and intracytoplasmic type A virus particles. Penetration of type B particles into cytoplasm of tumor cells has been observed. It consisted of inclusion of the particles into pinocytotic vesicles and of the release of the nucleoids into the cytoplasmic matrix. They were frequently seen penetrating into tumor cells producing type C or intracytoplasmic type A particles at the same time (Figs. 1 and 2). A similar penetration of other viruses into cells in vitro has been described as viropexis.

scholarly journals Evaluation of the in vivo effect of argon plasma coagulation with prior submucosal injection (Hybrid APC) in the esophagus of pigs

2019 ◽  
Author(s):  
Satoshi Abiko ◽  
Yuichi Shimizu ◽  
Shunsuke Ohnishi ◽  
Marin Ishikawa ◽  
Kana Matsuda ◽  
...  
Keyword(s):  
Argon Plasma Coagulation ◽  
Argon Plasma ◽  
Immunohistochemical Analysis ◽  
Type A ◽  
Muscle Layer ◽  
Type B ◽  
Submucosal Injection ◽  
Plasma Coagulation ◽  
Atrophic Muscle

Abstract Background Although argon-plasma coagulation (APC) is useful for treating early gastrointestinal cancer, safer ablation for oesophageal cancer is needed because the esophageal wall is very thin. The efficacy of APC with prior submucosal injection of saline (hybrid APC) by using a resected oesophagus of pig has been reported, but there has been no study in which the effects, biological reactions and delayed adverse effects of hybrid APC were evaluated. In this study, we evaluated the histological efficacy of APC with prior submucosal injection of saline (hybrid APC) by using an in vivo porcine model. Methods APC alone and hybrid APC were performed. Various settings of argon were used. The pigs were sacrificed after treatment (study 1) and 1 week after treatment (study 2). Histological evaluation of the deepest spot of coagulation from the basal layer (study 1) and non-atrophic muscle zone (study 2) in resected specimens was performed. Type A damage was defined as superficial tissue damage of the tunica mucosa, whereas type B damage was defined as an injury pattern limited to the tunica muscularis. The depths of type A and type B damage were measured in study 1. Immunohistochemical analysis was also performed in study 2. Results (study 1) Hybrid APC except for that at an excessive setting could prevent type B damage of the muscle layer. Standard APC at any setting could not prevent type B damage of the muscle layer. Results (study 2) The non-atrophic muscle zone was significantly larger in the hybrid APC group. Immunohistochemical analysis showed that the numbers of activated myofibroblasts and infiltrating neutrophils and macrophages were significantly smaller in the hybrid APC group than in the standard APC group. Conclusion APC following submucosal injection of saline contributes to sufficient and safe coagulation for oesophageal lesions.

scholarly journals Transcriptional Organization of the Region Encoding the Synthesis of the Flagellar Filament in Pseudomonas fluorescens

2008 ◽  
Vol 190 (11) ◽  
pp. 4106-4109 ◽  
Author(s):  
Miguel Redondo-Nieto ◽  
Javier Lloret ◽  
Javiera Larenas ◽  
Emma Barahona ◽  
Ana Navazo ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Sigma Factor ◽  
Type A ◽  
Type B ◽  
Master Regulator ◽  
Flagellar Filament

ABSTRACT Pseudomonas fluorescens F113 is motile by means of type b flagella. Analysis of the region encoding the synthesis of the flagellar filament has shown a transcriptional organization different from that of type a flagella. Additionally to the promoters driving fliC, fliD, and fleQ expression, we have found promoters upstream of the flaG gene and the fliST operon. These promoters were functional in vivo. Both promoters have been mapped and appear to be dependent on the vegetative sigma factor and independent of FleQ, the master regulator of flagellum synthesis.

Nucleosomes are assembled into discrete size structures by histone H1 in vitro

1986 ◽  
Vol 64 (5) ◽  
pp. 463-473 ◽  
Author(s):  
Teni Boulikas
Keyword(s):  
Ionic Strength ◽  
Histone H1 ◽  
Higher Order ◽  
Third Order ◽  
Calf Thymus ◽  
Exogenous Histone ◽  
Structural Subunit ◽  
Chromatin Structural ◽  
Low Ionic Strength

The involvement of histone H1 in the formation and maintenance of higher order chromatin structures in vitro was investigated biochemically. Addition of exogenous histone H1 to isolated calf thymus mononucleosomes in low ionic strength buffer resulted in the formation of electrophoretically distinct mononucleosome assemblies (supernucleosomes). The smaller super-nucleosomes were composed of about 12, 18, 24, or 30 nucleosomes and one to two molecules of histone H1 per nucleosome. It was difficult to determine accurately the size of the larger supernucleosomes, but their bands from native gels contained probably between 60 and 300 nucleosomes or more. Similar supemucleosome size classes were also obtained when oligonucleosomes instead of mononucleosomes were employed. When the assembly of mono- and oligo-nucleosomes with histone H1 took place in 0.15 M NaCl, discrete supernucleosomes containing only mono- or di-nucleosomes, but not a mixture of both, were formed. It is proposed that the small supernucleosomes containing oligomers of 6 nucleosomes may represent integral multiples of the second-order chromatin structural subunit, whereas the larger supernucleosomes containing about 60 to 300 or more nucleosomes may correspond to chromatin domains or third-order chromatin structures observed by other techniques.

Ventilatory changes during exercise and arterial PCO2 oscillations in chronic airway obstruction patients

1985 ◽  
Vol 58 (6) ◽  
pp. 1942-1948 ◽  
Author(s):  
J. G. Prior ◽  
M. Powlson ◽  
G. M. Cochrane ◽  
C. B. Wolff
Keyword(s):  
Airway Obstruction ◽  
Minute Ventilation ◽  
Type A ◽  
Fast Response ◽  
Type B ◽  
Early Exercise ◽  
Arterial Ph ◽  
Chronic Airway

Ventilatory kinetics during exercise (30 W for 6 min) were studied in 3 asthmatics, 14 patients with chronic airway obstruction (11 with bronchial or type B disease, 3 with emphysematous or type A disease), and in 5 normal age-matched controls. The measure of ventilatory increase during early exercise, alpha 1–3%, was calculated as (avg minute ventilation over 1st-3rd min of exercise--resting minute ventilation)/(avg minute ventilation over 4th-6th min of exercise--resting minute ventilation) X 100. Arterial pH, PO2, and PCO2 (PaCO2) were measured in vitro at rest and within 20 s of termination of exercise. Respiratory PaCO2 oscillations had previously been monitored at rest in the patients (indirectly as in vivo arterial pH, using a fast-response pH electrode) and quantified by upslope (delta PaCO2/delta t). alpha 1–3% was normal in asthmatics (whose respiratory oscillations as a group showed least attenuation) and in type A patients (whose respiratory oscillations as a group were most attenuated). In type B patients reduction in alpha 1–3% correlated with attenuation of delta PaCO2/delta t (r = 0.75; P less than 0.01). There was no significant correlation between delta PaCO2/delta t and change of in vitro PaCO2 from rest to the immediate postexercise period. These findings are consistent with the hypothesis that attenuation of delta PaCO2/delta t slows ventilatory kinetics during exercise in type B but not type A patients. Intact respiratory oscillations are not necessary for CO2 homeostasis after the first few minutes of exercise.

Biological and Morphological Studies on Low-Leukemia-Strain Mice with Hodgkin's-Like Neoplasia Induced by Serial Cell-Free Transmission of Leukemic Organs of SJL/J Strain Mice

1968 ◽  
Vol 26 ◽  
pp. 210-211
Author(s):  
S. Fujinaga ◽  
K. Maruyama ◽  
C.W. Williams ◽  
K. Sekhri ◽  
L. Dmochowski
Keyword(s):  
Tissue Culture ◽  
Type A ◽  
Reticulum Cell ◽  
Type B ◽  
Viral Origin ◽  
Serial Transfer ◽  
Strain Mouse ◽  
Morphological Studies ◽  
Cell Neoplasm ◽  
Free Material

Yumoto and Dmochowski (Cancer Res.27, 2098 (1967)) reported the presence of mature and immature type C leukemia virus particles in leukemic organs and tissues such as lymph nodes, spleen, thymus, liver, and kidneys of SJL/J strain mice with Hodgki's-like disease or reticulum cell neoplasm (type B). In an attempt to ascertain the possibility that this neoplasia may be of viral origin, experiments with induction and transmission of this neoplasm were carried out using cell-free extracts of leukemic organs from an SJL/J strain mouse with spontaneous disease.It has been possible to induce the disease in low-leukemia BALB/c and C3HZB strain mice and serially transfer the neoplasia by cell-free extracts of leukemic organs of these mice. Histological examination revealed the neoplasia to be of either reticulum cell-type A or type B. Serial transfer is now in its fifth passage. In addition leukemic spleen from another SJL/J strain mouse with spontaneous reticulum cell neoplasm (type A) was set up in tissue culture and is now in its 141st serial passage in vitro. Preliminary results indicate that cell-free material of 39th tissue culture passage can reproduce neoplasia in BALB/c mice.

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