Purinergic and mechanical interactions between myo- and secretory epithelial cells in mammary gland.
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Animals↗
Biomedical subjects
K Furuya
Publications and source records attributed to K Furuya.
Construction of high-resolution physical maps from yeast artificial chromosomes using restriction landmark genomic scanning (RLGS).
We have established a new system for chromosome-specific yeast artificial chromosome (YAC) contig construction using restriction landmark genomic scanning (RLGS-based YAC contig mapper). RLGS is a powerful tool for detecting more than 1000 restriction landmarks distributed on an entire genome in one procedure. In this system, RLGS is applied to sorted chromosomes to cover the target chromosome. Using these landmarks as guideposts, chromosome-specific YAC clones are then ordered. In this paper, we report the construction of a map for a human chromosome 21 YAC contig spanning q22.1 using this new approach. Applying RLGS to sorted chromosomes 21 enables detection of approximately 1400 spots (equivalent of 1050 PacI landmarks), covering the entire region of this chromosome. We constructed the 2.5-Mb YAC contig encompassing 21q22.1 with 66 spots (equivalent of 50 PacI landmarks). With this contig map, we could detect two deleted regions and chimerism in the YAC insert DNA. Our results demonstrated the usefulness of this approach for finding DNA alterations of YACs, such as deletions and chimerism.
Chimera↗
Upregulated expression of chemokines in herniated nucleus pulposus resorption.
STUDY DESIGN: Immunohistologic examination was performed on surgically removed samples of herniated nucleus pulposus. OBJECTIVES: To determine what cell types predominate in the granulation tissues of herniated nucleus pulposus, and to elucidate whether chemokines are involved in the resorption process of herniated nucleus pulposus. SUMMARY OF BACKGROUND DATA. The study population consisted of 30 patients suffering from herniated nucleus pulposus. Five macroscopically normal discs were obtained from spinal cord tumor and spinal cord injury managed with anterior discectomy (age range, 27-63 years) as a healthy control group. METHODS: Immunohistochemical analysis was used to analyze the expression of chemokines. RESULTS: A marked infiltration of macrophage and vascular proliferation was identified with a T lymphocyte infiltration of mild degree in the granulation tissues. This tendency was more prominent in the exposed group compared with the nonexposed group. Infiltrating macrophages, fibroblasts, and endothelial cells in the granulation tissues strongly expressed monocyte chemotactic protein-1 and macrophage inflammatory protein-1 alpha. Statistical analysis demonstrated that the exposed group was more abundant in Factor VIII, monocyte chemotactic protein-1, and macrophage inflammatory protein-1 alpha positive cells than the unexposed group. CONCLUSIONS: Inflammatory cells and their positivity for chemokines, such as monocyte chemotactic protein-1 and macrophage inflammatory protein-1 alpha, are associated with blood vessels. Chemokines, such as monocyte chemotactic protein-1 and macrophage inflammatory protein-1 alpha, were overexpressed in macrophages, fibroblasts, and endothelial cells, suggesting that these chemokines contribute to activation and recruitment of macrophages in a paracrine or autocrine fashion.
Adolescent↗
The natural history of herniated nucleus pulposus with radiculopathy.
STUDY DESIGN: The present study retrospectively investigated the morphologic changes that occurred during conservative treatment of patients with unilateral leg pain resulting from herniated nucleus pulposus without significant lumbar canal stenosis. OBJECTIVES: The results were correlated with clinical outcomes and extruding forms to determine which type of herniated nucleus pulposus had the greatest capacity for spontaneous regression and how rapidly such regression might occur. SUMMARY OF BACKGROUND DATA: The study population consisted of 77 patients with radiculopathy. METHODS: All patients complained primarily of unilateral leg pain, and 94% had positive tension signs. Additionally, 32% exhibited muscle weakness corresponding to the symptomatic nerve root. All patients were studied more than twice using magnetic resonance imaging during conservative therapy at a mean interval of 150 days. Morphologic changes on magnetic resonance imaging fell into four categories, with herniated nucleus pulposus classified into three types using T1-weighted sagittal views. Each patient was reexamined on the same scanner; 53 patients were examined twice, and 24 patients were examined more than three times. RESULTS: Morphologic changes, with the exception of 13 false-negative cases, basically corresponded to clinical outcome. In half of the cases that showed some improvement at follow-up evaluation, improvement of clinical findings were seen before those observed on magnetic resonance imaging. Migrating herniated nucleus pulposus frequently presented an obvious decrease in size, and even disappearance in seven cases. The further the herniated nucleus pulposus migrated, the more decrease in size could be observed. The cases apparently corresponding to "protrusion" showed little or no change on follow-up magnetic resonance imaging. Regarding the mechanism of herniated nucleus pulposus disappearance, exposure to the vascular supply undoubtedly took a part, although many factors were suspected to have some influence. CONCLUSION: Morphologic changes on magnetic resonance imaging mainly corresponded to clinical outcomes but tended to lag behind improvement of leg pain. Disappearance of herniate nucleus pulposus was seen frequently in the cases of migrating disc herniation, and it was presumed that exposure to the vascular supply had a lot to do with this phenomenon.
Adolescent↗
A long-range physical map of human chromosome 21q22.1 band from the YAC continuum.
The human Chromosome (Chr) 21q22.1 region contains several genes for cytokines and neurotransmitters and the gene for superoxide dismutase (mutant forms of which can cause familial amyotrophic lateral sclerosis). A region of approximately 5.8 Mb encompassing D21S82 and the glycinamide ribonucleotide transformylase (GART) loci was covered by overlapping YAC clones, which were contiguously ordered by clone walking with sequence-tagged site (STSs). A total of 76 markers, including 29 YAC end-specific STSs, were unambiguously ordered in this 5.8-Mb region, and the average interval between markers was 76 kb. Restriction maps of the YAC clones with rare-cutting enzymes were simultaneously prepared, and the restriction sites were aligned to obtain a consensus restriction map of the proximal region of the 21q22.1 band. The restriction map made from 44 overlapping YACs contains 54 physically assigned STSs. By integrating the consensus map of the adjacent 1.8-Mb region, we obtained a fine physical map spanning 6.5 Mb of human Chr 21q22.1. This map contains 24 precisely positioned end-specific STSs and 12 NotI-linking markers. More than 39 potential CpG islands were identified in this region and were found to be unevenly distributed. This physical map and the YACs should be useful as a reference map and as a resource for further structural analysis of the Giemsa-negative band (R-band) of Chr 21q22.1.
Base Sequence↗
Primary cavernous sinus malignant lymphoma treated by gamma knife radiosurgery: case report and review of the literature.
BACKGROUND: Malignant lymphomas originating primarily in the cavernous sinus have not been histologically verified by any authors. The first reported case to our knowledge of primary cavernous sinus malignant lymphoma, initially diagnosed as benign meningioma and treated by gamma knife radiosurgery, is presented. CASE DESCRIPTION: The patient was a 77-year-old man whose initial symptoms were left facial hypesthesia and diplopia on left gaze. During the 21-month follow-up period after gamma knife radiosurgery, no evidence of tumor regrowth was seen in the irradiated area, but an unirradiated mass expanded with invasion of the brain stem. Subsequent surgery provided histologic verification of the diagnosis. Despite successful local tumor control, the patient died due to sepsis 31 months after the onset of symptoms. Autopsy failed to disclose any remaining lymphoma cells either in the primary lesion or anywhere throughout the entire body. CONCLUSION: Primary cavernous sinus malignant lymphoma is an extremely rare brain tumor, but it must be considered in the differential diagnosis of an enhanced mass in the cavernous sinus. Stereotactic radiosurgery using the gamma knife technique for intracranial brain tumors, especially in the cavernous sinus, is associated with risk, as was seen in our patient. Our experience points out the pitfalls of gamma knife radiosurgery; it should be performed only after histologic confirmation has been obtained.
Aged↗
Forefoot pressures during walking in feet afficted with hallux valgus.
Forefoot pressures during walking in feet with hallux valgus were recorded using pressure-sensitive film, and the relationships between deformities and foot pressures were analyzed. There were 32 female subjects comprising 50 feet with hallux valgus, of which 20 feet underwent surgery. As in normal feet, the pressure patterns of feet with hallux valgus were varied: peak pressures were under the first metatarsal head, under the second and/or third metatarsal heads, and first, second, and/or third metatarsal heads. Peak pressures of hallux valgus feet were larger in each type than those of the normal foot. In feet with hallux valgus showing peak pressures under the first metatarsal heads, the hallux valgus angle and the intermetatarsal angle were larger than those in feet showing peak pressures on the second and/or third metatarsal heads. After surgery, peak pressures were under the second and/or third metatarsal heads and they decreased.
Adult↗
Intramedullary angiographically occult vascular malformations of the spinal cord.
OBJECTIVE: This study was performed to elucidate the differences between angiographically occult vascular malformations of the spinal cord and the histological subtypes. METHODS: The cases of six patients with spinal intramedullary angiographically occult vascular malformations encountered during the past 10 years were examined regarding clinical course, neuroradiological manifestations, and surgical consideration. RESULTS: There were four cavernous malformations and two arteriovenous malformations. The cervical section of the spinal cord was involved in four cases, and the thoracic section of the spinal cord was involved in two. All of the patients had presented with motor weakness and sensory disturbance below the level of the lesion. Two distinct clinical courses were observed; they were neurological decline with repeated episodes of relapse and remission (four patients) and continuing neurological decline (two patients). Although a mixed-intensity core surrounded by a low-intensity rim is thought to be characteristic of cavernous malformations, we were not able to differentiate the histological types on the basis of magnetic resonance imaging findings alone. CONCLUSION: In cases of spinal intramedullary angiographically occult vascular malformations, even if clinical course and magnetic resonance imaging findings are consistent with cavernous malformation, other histological subtypes need to be considered. Surgery should be considered for symptomatic patients, because symptoms and signs that are probably caused by bleeding tend to worsen rather than stabilize.
Adolescent↗
Correlation between the stimulatory effect of oxytocin on the formation of inositol phosphates and the oxytocin receptor level in the pregnant rabbit myometrium.
OBJECTIVE: In order to elucidate the roles of inositol trisphosphate (IP3) and oxytocin (OT) receptors in rabbit parturition, the concentration of IP3 induced by OT and the OT receptor levels were determined in rabbit myometria before and after parturition. METHODS: The effects of OT on IP3 formation and OT receptor levels were determined in the myometria of non-pregnant rabbits, Days 26, 28 and 30 of pregnancy rabbits, postpartum rabbits within 12 hours and steroid-treated ovariectomized rabbits. Prostaglandins (PGs) levels were also measured in the myometrial and decidual tissues. RESULTS: OT receptors were not detectable in the myometria of non-pregnant rabbits, and OT had no effect on the formation of inositol phosphates (IPs). On Day 28 of pregnancy, OT receptors became detectable, and then OT could induce the formation of IPs. Thereafter, the stimulatory effects of OT on IPs formation and the OT receptor levels dramatically increased toward the end of pregnancy and reduced rapidly after parturition. When the ovariectomized pregnant rabbits were treated with estrogen, OT receptors in the myometrium were induced, and OT acquired the ability to stimulate IP3 formation. However, OT had no effect on the production of cAMP, cGMP, prostaglandin (PG) E2 and F2 alpha in the myometria, even if receptors existed, although PGE2 production in the decidual tissues was markedly stimulated. In addition, an OT receptor antagonist inhibited the stimulatory effects of OT on IP3 formation. CONCLUSIONS: These results suggest that the formation of IPs by OT, the OT receptor levels in the myometrium, and the production of PGF2 in the decidua might play crucial roles in parturition.
Animals↗
The Streptomyces peucetius drrC gene encodes a UvrA-like protein involved in daunorubicin resistance and production.
The drrC gene, cloned from the daunorubicin (DNR)- and doxorubicin-producing strain of Streptomyces peucetius ATCC 29050, encodes a 764-amino-acid protein with a strong sequence similarity to the Escherichia coli and Micrococcus luteus UvrA proteins involved in excision repair of DNA. Expression of drrC was correlated with the timing of DNR production in the growth medium tested and was not dependent on the presence of DNR. Since introduction of drrC into Streptomyces lividans imparted a DNR resistance phenotype, this gene is believed to be a DNR resistance gene. The drrC gene could be disrupted in the non-DNR-producing S. peucetius dnrJ mutant but not in the wild-type strain, and the resulting dnrJ drrC double mutant was significantly more sensitive to DNR in efficiency-of-plating experiments. Expression of drrC in an E. coli uvrA strain conferred significant DNR resistance to this highly DNR-sensitive mutant. However, the DrrC protein did not complement the uvrA mutation to protect the mutant from the lethal effects of UV or mitomycin even though it enhanced the UV resistance of a uvrA+ strain. We speculate that the DrrC protein mediates a novel type of DNR resistance, possibly different from the mechanism of DNR resistance governed by the S. peucetius drrAB genes, which are believed to encode a DNR antiporter.
Adenosine Triphosphatases↗
The DnrN protein of Streptomyces peucetius, a pseudo-response regulator, is a DNA-binding protein involved in the regulation of daunorubicin biosynthesis.
DnrN, a protein essential for the transcription of the dnrI gene, which in turn activates transcription of the daunorubicin biosynthesis genes in Streptomyces peucetius, was overproduced in Escherichia coli and S. peucetius. The cell-free extract from E. coli was used to conduct DNA-binding assays. The results of gel mobility shift analysis showed that DnrN binds specifically to the dnrI promoter region with a high affinity (Kd = 50 nM). Neither acetyl phosphate nor ATP affected the binding ability, and there was no difference in binding between wild-type DnrN and a mutant form (D-55-->N) lacking the putative phosphorylation site (aspartate 55) of a response regulator protein. Therefore, phosphorylation of DnrN apparently is not necessary for DNA binding. DNase I footprinting analysis indicated binding regions at 37 to 55 bp and 62 to 100 bp upstream of the transcriptional start point of dnrI. Interestingly, the sequence of these regions includes consecutive overlapping triplets [5'-(A/T)GC, 5'-(A/T)CG, 5'-(A/T)C(A/T)] that have been shown to be the preferential binding site of daunorubicin (J. B. Chaires and J. E. Herrera, Biochemistry 29:6145-6153, 1990). This may explain why daunorubicin appeared to inhibit the binding of DnrN to the dnrI promoter, which could result in feedback repression of daunorubicin production. The results of Western blotting (immunoblotting) analysis with His-tagged DnrN antiserum showed that dnrN expression is coincident with daunorubicin production and that the maximum level of DnrN is 0.01% of total protein in the wild-type S. peucetius strain. Since the level of DnrN was lowered in mutant strains that do not produce daunorubicin, we speculate that dnrN and dnrI expression are regulated by daunorubicin.
Bacterial Proteins↗
Deletion polymorphism in the angiotensin I converting enzyme (ACE) gene as a genetic risk factor for sarcoidosis.
BACKGROUND: Genetic control of serum angiotensin I converting enzyme (SACE) levels has been suggested. A study was undertaken to elucidate the role of this polymorphism in sarcoidosis. METHODS: Three hundred and forty one unrelated healthy controls and 103 consecutive patients with sarcoidosis participated in the study. SACE levels and an insertion/deletion (I/D) polymorphism in intron 16 of the ACE gene were studied in each subject and new reference intervals for SACE activity for each genotype were determined. The difference in genotype and allele frequencies between controls and patients was analysed and odds ratios were calculated to estimate the relative risk. RESULTS: A significant association was seen between ACE gene polymorphism and SACE levels in both patients and controls. The new reference intervals for each genotype discriminated abnormal SACE levels in patients more accurately, especially those with genotype II. In women the frequencies of allele I were 0.68 (allele D 0.32) in controls and 0.58 (allele D 0.42) in patients, and the difference between the two female groups was significant (p < 0.05). Thus, an excess of genotype ID or DD was observed in female patients (odds ratio 2.18; 95% confidence interval 1.18 to 4.01; p = 0.01). CONCLUSIONS: These findings suggest that ACE gene polymorphism is associated with SACE levels in both patients with sarcoidosis and controls. ACE gene polymorphism should be further evaluated as a candidate marker for an increased risk of sarcoidosis.
Adult↗
Effect of culture supernatants of endometriotic lesions, uterine endometrium and peritoneum from rats with experimental endometriosis on the natural killer activity of spleen cells.
Experimental endometriosis in rats was induced by autotransplanting the uterine endometrium to the peritoneum. In all rats, endometrial implants developed into endometriotic tissues similar to those in humans about 2 weeks after transplantation. Natural killer (NK) activity of spleen cells in the endometriosis model rats was significantly (p < 0.05) lower than that in the sham-operated intact rats. The inhibited NK activity in the endometriosis rats recovered to the level in intact rats with danazol (but not buserelin). The supernatant after 24-hour culture of endometrial tissues from both intact and model rats seemed to have significant inhibitory effects on NK activity. The supernatant from endometrial grafts showed significantly (p < 0.05) higher inhibitory effects than that from the endometrial tissues. The inhibitory effects were significantly (p < 0.05) reduced by treatment with danazol or buserelin to the untreated level. In addition, supernatants of unaffected peritoneal tissues from the endometriosis rats had significantly (p < 0.01) higher inhibitory effects on NK activity than those from the intact rats. Even when uterine serosa or silicone was implanted to the peritoneum, the supernatants of the contralateral peritoneal tissues showed significantly (p < 0.05) higher inhibitory effects than those from the intact rats, while having significantly (p < 0.05) lower inhibitory effects than those from the endometriosis rats. These results suggest that this marked inhibitory effect on NK activity by the peritoneum may be associated with the development and progression of endometriosis.
Animals↗
Intracellular free calcium responses during chemotaxis of Dictyostelium cells.
A calcium ion indicator, fura-2 bovine serum albumin, was introduced into Dictyostelium discoideum cells by electroporation. The concentration of intracellular calcium ions ([Ca2+]i) increased transiently in vegetative cells upon stimulation with submicromolar concentrations of folic acid, a chemoattractant for this organism at the vegetative stage. Similar [Ca2+]i responses were also observed in aggregation-competent cells upon stimulation with subnanomolar concentrations of cAMP, a chemoattractant at the aggregation stage. The [Ca2+]i response caused by cAMP was 2.1 times higher than that caused by folic acid. The magnitude of these responses depended on the concentration of Ca2+ in the external buffer. The presence of magnesium ions inhibited the [Ca2+]i responses in a dose-dependent manner. [Ca2+]i was higher in the rear region than in the anterior region of cells freely migrating on the surface, although such a gradient was not always maintained. When aggregation competent cells were locally stimulated by the application of a microcapillary containing cAMP, the cells extended pseudopods toward the microcapillary. In these cases, an increase in [Ca2+]i was transiently observed in the region opposite to the tip of the capillary. At the slug stage, [Ca2+]i was higher in prestalk cells than in prespore cells of slugs. The possibility that the [Ca2+]i is spatially regulated within a cell was discussed.
Animals↗
Spontaneous rupture of the uterine vessels in a pregnancy complicated by endometriosis.
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Adult↗
1.8-megabases fine physical map encompassing IFNAR and AML1 loci on human chromosome 21q22.1.
A long-range restriction map of the 1.8-megabases (mb) region encompassing the area between the interferon-alpha receptor and the acute myelogenous leukemia loci on human chromosome 21q22.1 was constructed after analysis of both the contiguous yeast artificial chromosome (YAC) clones and genomic DNA. Analysis of pulsed-field gel electrophoresis of lymphoblastoid DNA digested with three rare-cutting enzymes, Not I, Mlu I, and Nru I, revealed the positions of 17 markers on each restriction map. The 1.8-mb YAC contig that covers this region was obtained through YAC walking mediated by sequence-tagged sites (STSs), with 29 STSs including 12 newly generated YAC end-specific STSs. The consensus restriction map from 15 overlapping YACs and the positioning of the STS markers on each clone allowed 24 markers including 4 Not I-linking STSs to be ordered and mapped physically. Comparison of the maps revealed that the proximal region contains more unmethylated CpG islands than the distal region, which suggests that many expressed genes are in the proximal region. This fine consensus physical map will be informative and useful for construction of contigs of cosmid, P1, or BAC clones for further large-scale sequencing in this gene-rich region.
Base Sequence↗
Inverse relation between vertebral body deformity and intervertebral disk narrowing in lumbar spine of elderly women.
To investigate the relation between vertebral body deformity and disk narrowing in the lumbar spine of elderly patients, a longitudinal radiographic study of 56 aged women was retrospectively performed. We measured disk areas and body heights on two lateral lumbar radiograph sets obtained with an average interval between examinations of 10 years. The absolute values of disk areas on first examination and the rates of change of area of disks adjacent to nondeformed bodies were smaller than the corresponding values for the deformed bodies. Conversely, deformed bodies tended to have normal or expanded disks regardless of the mode of deformity. Despite the small number of subjects studied, our findings suggested that an inverse relation may exist between vertebral body deformity and disk narrowing.
Aged↗
[Research trends in molecular genetics of respiratory medicine].
In order to identify necessary or susceptibility genes of respiratory diseases, molecular genetic approaches have been chosen more frequently than before because a tremendous number of anonymous markers over the human genome are now available. However, conventional linkage analysis such as the maximum likelihood method and affected sib-pair analysis have limitations mainly due to heterogeneity of disease genes. Association studies have more power to detect susceptibility genes, however, they tend to yield more false positive results than linkage studies unless corrected for multiple observations. There are some new methods potentially applicable to a genome-wide search for candidate genes. Each method has advantages and disadvantages. Reverse genetics is promising but destined to return to regular biochemical, physiological, and clinical approaches of respiratory medicine.
Disease Susceptibility↗