PubMed Health⌕ Search

Biomedical subjects

T Akitaya

Publications and source records attributed to T Akitaya.

12 recordsLinked to original sources
PubMed2001

Intercalating fluorescence dye YOYO-1 prevents the folding transition in giant duplex DNA.

Recently, it has become clear that with the addition of polyamines, giant DNA molecules of size greater than 10 kbp exhibit all-or-none switching between elongated coil and folded compact states. Here the effects of the intercalating fluorescent labeling dye, YOYO-1, and the minor-groove binding fluorescent labeling dye, DAPI, on the folding transition of single giant T4 DNA (166 kbp) induced by spermidine(3+) were examined, by use of the experimental technique of single molecular chain observation with fluorescence microscopy. It is found that the intercalating dye, YOYO-1, markedly prevents the folding transition, whereas the minor-groove binding dye, DAPI, exhibits negligible effect on the folding transition. This action of YOYO-1 is discussed in relation to the biological effect of intercalators.

Benzoxazoles↗
PubMed1998

Detection of regional lymph node metastases in colon cancer by using RT-PCR for matrix metalloproteinase 7, matrilysin.

Lymph node metastasis is the most important prognostic factor in colon cancer. However, more accurate screening for metastasis than that afforded by conventional pathology remains elusive. We have employed a reverse transcriptase-polymerase chain reaction (RT-PCR) assay for a matrix metalloproteinase (MMP), 'matrilysin', because this gene is epithelial-specific and consistently expressed in colorectal cancer cells. The sensitivity of this assay was examined with the matrilysin-producing rectal cancer cell line 'CaR-1'. Matrilysin mRNA was detected in this system when more than 10(4) matrilysin-positive cells existed in a lymph node of ordinary size. Fourteen of 15 (93%) primary colon cancers and none of the surrounding normal tissues expressed matrilysin. All 10 histologically-positive lymph nodes were positive for matrilysin, while of 60 histologically-negative lymph nodes, eight were positive for matrilysin. When the additional sequential sectioning and histological re-examination was performed on five of these eight 'matrilysin-positive, but histologically-negative' lymph nodes, micrometastases were detected in three. Only one of the lymph nodes that were histologically-positive, but negative by matrilysin assay was from a patient with colon cancer in which matrilysin was not detected. In conclusion, RT-PCR assay for matrilysin is a sensitive method for detecting occult metastases in patients with colon cancer, and may complement histologic examination.

Adult↗
PubMed1997

Construction of cDNA bank from biopsy specimens for multiple gene analysis of cancer.

We constructed a "cDNA bank" of human colorectal cancer and surrounding normal tissues with our unique mRNA assay system. Total nucleic acids extracted from patients' tissues were applied to 96-well plates, where poly(dT) sequences of oligonucleotides were immobilized. After hybridization, the cDNA was reverse-transcribed on the plate with the captured mRNA as a template, followed by synthesis of double-stranded (ds) cDNA. The resulting sense cDNA was removed from the plate, then used in PCR for analysis of various genes. The sense strand of the cDNA was repeatedly synthesized by using the immobilized antisense cDNA as a template even from plates used once and stored at 4 degrees C for as long as 6 months. Furthermore, the results of PCR could be easily compared among different specimens if the same amount of total mRNA were applied to the plate for the ds cDNA synthesis. This demonstrated that the cDNA bank constructed from clinical materials provides almost unlimited supplies of cDNA for multiple gene analysis of cancer.

Actins↗
PubMed1996

Matrilysin is associated with progression of colorectal tumor.

Matrilysin and gelatinase A, B mRNA expressions were examined in colorectal tumors. Matrilysin mRNA was observed exclusively in tumors, while the others were also found in normal mucosa surrounding tumors. Further analysis revealed that colorectal adenomas with severe dysplasia, not with mild dysplasia, expressed matrilysin with lower levels than cancers. The level of matrilysin mRNA expression increased with the advancement of stages of colorectal cancers, consequently a relatively higher expression was observed in liver metastatic tumors than primary tumors. These results suggest that matrilysin mRNA expression was correlated with the progression of colorectal tumors, and this enzyme may also play a role in developing metastatic tumors in liver.

Adenoma↗
PubMed1995

Keratinocytes constitutively express the Fas antigen that mediates apoptosis in IFN gamma-treated cultured keratinocytes.

The Fas antigen is a cell surface protein that can mediate apoptosis in many cell types. Although its physiological function is still unclear, recent evidence indicates that this surface molecule is involved in apoptosis in the immune system and the liver. The epidermis is an organ that undergoes terminal differentiation with the eventual death of keratinocytes, and it has been suggested that this is a specialized form of apoptosis. In the present study, we examined whether or not the Fas antigen is involved in keratinocyte apoptosis. Immunoreactivity for the Fas antigen was found throughout the epidermis in normal human skin sections and cultured normal human keratinocytes, and mRNA for the Fas antigen was found to be constitutively expressed in normal epidermis and cultured normal keratinocytes by RT-PCR analysis. To determine whether the Fas antigen in keratinocytes is functional, we used a cytotoxic monoclonal antibody (mAb) against the Fas antigen to induce apoptosis. This antibody did not induce apoptosis of cultured keratinocytes even though they expressed the Fas antigen. We then tested the ability of several cytokines (TGF beta, TNF alpha and IFN gamma) to induce Fas-mediated keratinocyte apoptosis. Only pretreatment with IFN gamma followed by the addition of the anti-Fas mAb induced apoptosis, as assessed by cell viability, morphological changes and ultrastructural characteristics, suggesting that constitutive expression of the Fas antigen is not sufficient to induce apoptosis in keratinocytes and that keratinocyte apoptosis via the Fas antigen-mediated mechanism may require the activation of keratinocytes by IFN gamma, which is thought to be produced by activated T cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1992

Expression of cell adhesion molecules during initiation and cessation of neural crest cell migration.

Because of their distribution and known ability to promote neuronal adhesion, it has been proposed that N-CAM and N-cadherin are involved in the formation of the nervous system. Here, we examine the expression of these molecules during the initiation and cessation of trunk neural crest cell migration during the formation of the peripheral nervous system. Whereas other neural tube cells express N-cadherin, the dorsal neural tube containing neural crest precursors has little or no N-cadherin immunoreactivity. In contrast, N-CAM is expressed in the dorsal neural tube and on early migrating neural crest cells, from which it gradually disappears during migration. Both N-CAM and N-cadherin are absent from neural crest cells at advanced stages of migration. As neural crest cells cease migration and condense to form dorsal root and sympathetic ganglia, N-cadherin but not N-CAM is observed on the forming ganglia, identified by neurofilament expression and the aggregation of HNK-1 reactive cells. The results demonstrate that the absence of N-cadherin correlates with the onset of neural crest migration and its reappearance correlates with cessation of migration and precedes gangliogenesis.

Animals↗
PubMed1991

Amyloid beta protein substituent peptides do not interact with the substance P receptor expressed in cultured cells.

The amyloid beta protein (ABP) has been shown to interact with the substance P (SP) receptor in a cell culture model that may mimic the pathogenesis of Alzheimer's disease. In the present study, however, 4 fragments of ABP (beta 1-42, beta 1-16, beta 17-28, and beta 25-35) failed to interact with SP-induced Ca2+ mobilization in SP receptor-expressing cultured cells. Therefore, the action of these ABP-related peptides in our cultured cells is unrelated to the SP receptor.

Alzheimer Disease↗
PubMed1986

Spatial and temporal organization of intracellular adenine nucleotides and cyclic nucleotides in relation to rhythmic motility in Physarum plasmodium.

Spatio-temporal organization of a migrating plasmodium was studied both by analysing intracellular concentrations of adenine and cyclic nucleotides and by applying image processing for recording oscillatory changes in thickness with use of microcomputers. ATP and ADP concentrations were about twice as high in the front as in the rear, while AMP distributed uniformly. On the other hand, cAMP and cGMP concentrations were several times higher in the rear than in the front, showing oscillations in between. The cAMP concentrations at the front oscillated with a phase advancing about one-third of the period with respect to the phase of the thickness oscillation, while cGMP concentration there varied only little. ATP concentration oscillated concomitantly with H+. A feedback control loop consisting of (ATP-H+)-cAMP-Ca2+ is proposed. The possible mechanism of rhythmic contractions involving mitochondria which may excrete pulses of Ca2+ and induce cell polarization is discussed.

Adenine Nucleotides↗
PubMedDate not supplied

Effect of microaerosol inhalation on the pattern of breathing.

Microaerosol stimulation of the trachea and bronchi appears to have no effect on the pattern of breathing. However, laryngeal stimulation with microaerosol produces consistent changes characterized by slowing of the inspiratory effort. The reflex response can be abolished by either bilateral superior laryngeal nerve section or topical anesthesia of the laryngeal mucous membranes. Nasal stiumulation with saline microaerosol results in marked slowing of inspiration. This response can also be blocked by topical anesthesia of the nasal mucosa. The decreased breathing rate results from both increased t insp. and t exp. During inspiration the build-up phrenic discharge activity, the phrenic slope, decreases. The phrenic peak, however, remains unchanged. The nose and larynx, therefore, appear to play significant roles in the modification of the breathing pattern in response to a nonirritating solution delivered as a microaersol. Slower breathing, relexly produced in the human being by such a method, might benefit partial airway obstruction by reducing turbulence and therefore the work of breathing. It is speculated that in a marginally competent upper airway, this benefit might be a critical factor in survival.

Aerosols↗