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Biomedical subjects

E Tani

Publications and source records attributed to E Tani.

At least 19 recordsLinked to original sources

Growth fraction in non-Hodgkin's lymphomas and reactive lymphadenitis determined by Ki-67 monoclonal antibody in fine-needle aspirates.

The fraction of proliferation cells was analysed in fine needle aspirates from a series of 448 non-Hodgkin's lymphomas and 199 reactive hyperplasias using an immunoperoxidase staining with monoclonal antibody Ki-67. There was a good correlation between proliferation fraction and cytologic assignment to high and low grade lymphomas. Thus high grade lymphomas had a high median percentage of Ki-67 positive cells with a figure of 82.1 for lymphoblastic, 60.0 for immunoblastic, and 59.7 for centroblastic lymphomas. For low grade lymphomas the figures were 17.1 and 11.1 percent for centroblastic/centrocytic and CLL/immunocytoma, respectively. The fraction of proliferation cells in reactive lymphadenitis varied between 1-50% with a median of 11.5%. Analysis of Ki-67 positivity can accordingly not be used to differentiate benign from neoplastic proliferations. Within all lymphoma subgroups but lymphoblastic lymphoma, there was a marked variation in fraction of Ki-67 positive cells, which resulted in a certain overlap between high and low grade lymphomas. The results show that cells procured through fine-needle aspiration can be used to analyse the fraction of proliferating cells which contributes information about the growth rate of the individual tumours that can not be obtained through cytologic classification.

Antibodies, Monoclonal

Three distinct phases of fodrin proteolysis induced in postischemic hippocampus. Involvement of calpain and unidentified protease.

BACKGROUND AND PURPOSE: Fodrin, a neuronal cytoskeleton protein, is proteolyzed by calpain after ischemic insult. We examined proteolysis of fodrin induced by global forebrain ischemia in gerbil hippocampus in spatial terms by using the antibody specific to the calpain-proteolyzed form of fodrin. METHODS: In gerbils, a 10-minute forebrain ischemia was produced by occlusion of both carotid arteries. After recirculation, the hippocampus was processed for immunohistochemical and immunoblot study with the antibody against the calpain-proteolyzed form of fodrin. Additionally, short-term ischemia was studied to find the threshold of fodrin proteolysis. RESULTS: Three phases of fodrin proteolysis distinct in chronology and distribution arose: (1) an early predegeneration phase in the molecular layer and stratum oriens of the CA1 and CA3 sectors within the first 15 minutes, which lasted up to 4 hours; (2) a late predegeneration phase in the whole CA1 sector, except for the pyramidal cells, between 12 hours and 2 days; and (3) a postdegeneration phase in the cytoplasm of the CA1 neurons, which arose in 3 to 7 days. A 4-minute (not a 3-minute) forebrain ischemia induced the late predegeneration phase of fodrin proteolysis and delayed neuronal death in CA1. Immunoblotting showed that the primary product of calpain action was further proteolyzed by an unidentified protease. CONCLUSIONS: Calpain induced proteolysis of fodrin in ischemic hippocampus, and the late predegeneration phase of the proteolysis was closely associated with the delayed neuronal death in the CA1 sector. Calpain and another protease may play a role in the development of neuronal death after transient forebrain ischemia.

Animals

Activity of smooth muscle phosphatases 1 and 2A in rabbit basilar artery in vasospasm.

BACKGROUND AND PURPOSE: Subarachnoid hemorrhage frequently leads to a long-term cerebral artery narrowing called vasospasm. Recently, the involvement of myosin light chain kinase has been found in experimental vasospasm in our laboratory. We therefore measured the activity of serine/threonine protein phosphatases 1 and 2A in the rabbit basilar artery in vasospasm and in vasocontraction to study their role, particularly in regard to vasospasm compared with vasocontraction. METHODS: Vasospasm was produced in the rabbit basilar artery by a two-hemorrhage method. Vasocontraction was induced by local application of KCl or serotonin to the rabbit basilar artery after a transclival exposure. The control animals were treated with saline instead of fresh blood. Serine/threonine protein phosphatase activity in the basilar artery was assayed with the use of [32P]phosphorylase-a as a substrate; protein phosphatase 1 activity was evaluated as protein phosphatase activity in the presence of 1 nmol/L okadaic acid, whereas protein phosphatase 2A activity was assessed as protein phosphatase activity inhibited by 1 nmol/L okadaic acid. RESULTS: Values of mean activity of protein phosphatase 1 in myofibrillar extract were 3.58 +/- 0.26 nmol/min per milligram in the control group, 3.22 +/- 0.12 nmol/min per milligram in the spastic group on day 2, and 3.01 +/- 0.16 nmol/min per milligram in the spastic group on day 4 (a significant decrease in protein phosphatase 1 activity in the spastic group on days 2 and 4). In contrast, these values did not show any significant changes in the KCl and serotonin groups. Values of mean activity of protein phosphatase 2A in cytosolic extract were 0.90 +/- 0.07 nmol/min per milligram in the control group, 0.75 +/- 0.10 nmol/min per milligram in the spastic group on day 2, and 0.62 +/- 0.17 nmol/min per milligram in the spastic group on day 4 (a significant reduction in protein phosphatase 2A in the spastic group on days 2 and 4). There was no evidence of significant changes of protein phosphatase 2A in cytosolic extract in the KCl and serotonin groups. CONCLUSIONS: Protein phosphatase 1 in myofibrillar extract is reported to catalyze the dephosphorylation of myosin light chain and calponin, whereas protein phosphatase 2A in cytosolic extract catalyzes the dephosphorylation of calponin and caldesmon. In addition, the phosphorylation of calponin and caldesmon results in the loss of their ability to inhibit smooth muscle contraction. Therefore, the significant decrease in activity of protein phosphatases 1 and 2A in vasospasm may result in uninterrupted vascular smooth muscle contraction by the preservation of phosphorylation of not only myosin light chain but also calponin and caldesmon.

Animals

Effects of protein kinase C modulators on multidrug resistance in human glioma cells.

To identify the role of protein kinase C (PKC) in multidrug resistance, the effects of phorbol-12-myristate-13-acetate (PMA), a PKC activator, or calphostin C, a PKC inhibitor, on intracellular vincristine accumulation and expression of P-glycoprotein phosphorylation were studied in one multidrug-resistant and three multidrug-sensitive human glioma cell lines. Basal PKC activities and immunoreactivities of PKC-alpha and -zeta were higher in multidrug-resistant cells than in multidrug-sensitive cells. There was no significant difference in the immunoreactivity of PKC-delta between multidrug-resistant and -sensitive cells, and immunoreactive PKC-beta, -gamma, and -epsilon were not detected in either multidrug-resistant or -sensitive cells. The treatment of multidrug-resistant cells with 100 nM PMA for 2 hours resulted in the activation not of PKC-zeta but of PKC-alpha, with concomitant decrease in vincristine accumulation and increase in P-glycoprotein phosphorylation. The exposure of multidrug-resistant cells to 100 nM PMA for 24 hours induced down-regulation not of PKC-zeta but of PKC-alpha, with concurrent decrease in vincristine accumulation, and reduced but still increased P-glycoprotein phosphorylation. The treatment of multidrug-resistant cells with 100 nM calphostin C for 2 hours decreased immunoreactive PKC-zeta and not immunoreactive PKC-alpha, inducing increase in vincristine accumulation, with concomitant decrease in P-glycoprotein phosphorylation. There was no evidence of significant change in vincristine accumulation in multidrug-sensitive cells treated with PMA or calphostin C. This may suggest that at least two isozymes of PKC, PKC-alpha and -zeta, are involved in P-glycoprotein phosphorylation and that vincristine efflux function in multidrug-resistant human glioma cells is closely associated with P-glycoprotein phosphorylation and is decreased by PKC inhibitor.

ATP Binding Cassette Transporter, Subfamily B, Mem

Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gliomas.

The gene expression of five matrix metalloproteinases (MMPs) and two tissue inhibitors of metalloproteinases (TIMPs) was studied in human gliomas in vivo and in vitro to evaluate their roles in glioma invasion. Simultaneous expression of one to four MMP genes and two TIMP genes was found in 17 surgical glioma specimens, and one MMP (gelatinase A) gene and two TIMP genes were simultaneously expressed in tissue of three brains. The concomitant overexpression of gelatinase A, gelatinase B, and occasional matrilysin genes was associated with the malignancy of gliomas and accompanied by overexpression of the TIMP-1 gene. In five human glioma cell lines, gelatinase A, TIMP-1, and TIMP-2 genes were constitutively expressed in alll cell lines: the matrilysin gene in three cell lines; the stromelysin gene in two cell lines; and the interstitial collagenase gene in one cell line. There was a clear difference in the expression of gelatinase B and stromelysin genes between surgical glioma specimens and glioma cell lines: the gelatinase B gene was not expressed constitutively in vitro but was overexpressed in vivo, whereas the stromelysin gene was not expressed in vivo but was expressed in some cell lines. To find the cause of that difference in vivo and in vitro, the transcriptional regulations of MMP and TIMP genes by tumor promoter, growth factors, or cytokines were studied in vitro. Interstitial collagenase, gelatinase B, stromelysin, and TIMP-1 genes were upregulated in many cell lines by phorbol-12-myristate-13-acetate (PMA) and in some cell lines by epidermal growth factor, tumor necrosis factor-alpha, or interleukin-1 beta. Transforming growth factor-beta 1 (TGF beta 1) upregulated gelatinase A and matrilysin genes in some cell lines, and there were no clear responses from any MMP and TIMP genes to interleukin-6. Thus, the transcriptional modulation of MMP genes by these growth factors and cytokines seemed insufficient to explain the difference in gelatinase B and stromelysin gene expressions in vivo and in vitro and was suggestive of the genetic alteration of glioma cells in vitro, the heterogeneous cell population in glioma tissues, or both. Furthermore, the in vitro invasion of glioma cells through Matrigel in response to PMA, TGF beta 1, or TIMP-1 was assessed by chemoinvasion assay. In most cell lines, invasion was significantly stimulated by PMA or TGF beta 1 but suppressed by TIMP-1.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult

Inhibition by 5'-methylthioadenosine of cell growth and tyrosine kinase activity stimulated by fibroblast growth factor receptor in human gliomas.

Stimulation of three human glioma cell lines with basic fibroblast growth factor (bFGF) led to the enhancement of cell growth and the rapid tyrosine phosphorylation of cellular proteins, including major substrates of 90 kD. A methyltransferase inhibitor, 5'-methylthioadenosine (MTA), inhibited dose dependently the bFGF-stimulated cell growth and protein tyrosine phosphorylation in glioma cells by blocking both receptor autophosphorylation and substrate phosphorylation, as shown by immunoblotting with antiphosphotyrosine antibodies and cross-linking bFGF to receptors. The antiproliferative activity of MTA correlated quantitatively with its potency as an inhibitor of bFGF-stimulated protein tyrosine kinase activity. The methyltransferase inhibitor MTA had no effect on either epidermal growth factor- or platelet-derived growth factor-stimulated protein tyrosine phosphorylation in glioma cells, but inhibited specifically bFGF-stimulated protein tyrosine kinase activity. The concentration of MTA required for inhibition of protein methylation correlated well with the concentration required for inhibition of bFGF-stimulated cell growth and protein tyrosine phosphorylation. Because MTA had no effect on numbers and dissociation constants of high- and low-affinity bFGF receptors, the inhibition of bFGF-stimulated bFGF receptor tyrosine kinase activity is not likely to be the result of a reduction in bFGF receptor and bFGF binding capacity. In fact, MTA delayed and reduced the internalization and nuclear translocation of bFGF, and the internalized bFGF was submitted to a limited proteolysis that converted it to lower molecular peptides whose presence remained for at least 22 hours. The effect of MTA on bFGF-stimulated tyrosine phosphorylation was immediate and readily reversible.

Antibodies

Apoptosis of human glioma cells in response to calphostin C, a specific protein kinase C inhibitor.

Calphostin C acts at the regulatory domain as a highly selective inhibitor of protein kinase C (PKC), and staurosporine acts at the catalytic domain as a nonspecific PKC inhibitor. The authors investigated the capacity of calphostin C and staurosporine to promote apoptotic fragmentation of DNA in four human glioma cell lines. The exposure of glioma cell lines to 100 nM calphostin C for 2 to 8 hours induced a decrease in particulate PKC activities and exposure for 16 to 24 hours produced a concentration-dependent increase in internucleosomal DNA cleavage on agarose gel electrophoresis. In addition, the human glioma cells showed the classic morphological features of apoptosis: cell shrinkage, nuclear condensation, and the formation of apoptotic bodies. A 24-hour exposure to staurosporine failed to induce internucleosomal DNA fragmentation at concentrations generally used to achieve maximum inhibition of enzyme activity (50 nM) but promoted fragmentation at considerably higher concentration (more than 200 nM). Deoxyribonucleic acid fragments obtained from cells exposed to 100 nM calphostin C for 16 to 24 hours possessed predominantly 5'-phosphate termini, consistent with the action of a Ca++/Mg(++)-dependent endonuclease. Northern and Western blot analyses revealed that the exposure to 100 nM calphostin C for 4 hours failed to alter bcl-2 transcript and protein, but exposure for more than 8 hours decreased the amount of bcl-2 transcript and protein. Together, these observations suggest that calphostin C is capable of inducing apoptotic DNA fragmentation and cell death in a highly concentration dependent manner in human glioma cells and that the apoptosis is closely associated with the decrease in transcription and translation of bcl-2.

Alkaloids

Effect of phenobarbital, spironolactone and pregnenolone-16 alpha-carbonitrile on rat hepatic beta-glucuronidase.

beta-Glucuronidase cleaves beta-glucuronides of drugs, steroids, as well as macromolecular compounds containing glucuronic acid bound with a beta-glycoside bond. Therefore, the study of induction of this enzyme by known microsomal enzyme inducers is of obvious interest. The effect of phenobarbital, spironolactone and pregnenolone-16 alpha-carbonitrle on the rat liver beta-glucuronidase was studied. These compounds augmented the activity of this enzyme, in the above increasing order. These results are discussed in view of the influence on pharmacokinetics of many drugs. Furthermore, they may lead to new therapeutic approaches for certain diseases.

Animals

[Increased expression of gelatinases A and B, matrilysin and TIMP-1 genes in human malignant gliomas ].

Malignant glioma is a local invasive tumor in the central nervous system. The mRNA expression of five matrix metalloproteinases (MMPs) and two tissue inhibitors of metalloproteinases (TIMPs) was examined in surgical specimens of three brain tissues, two astrocytomas, four anaplastic astrocytomas and eleven glioblastomas, including recurrent one anaplastic astrocytoma and two glioblastomas. In the control brain tissues, mRNA expression was high for TIMP-2, low for gelatinase A and TIMP-1, and undetectable for gelatinase B, interstitial collagenase, stromelysin and matrilysin. Gelatinase B and TIMP-1 were concomitantly overexpressed in primary glioblastomas. In addition, the average expression level of gelatinase A increased 3.0 fold in astrocytomas and anaplastic astrocytomas and 6.0 fold in glioblastomas, compared to the brain tissues. Matrilysin was induced variably in more than half of the primary glioblastomas, and interstitial collagenase was slightly induced in some primary and recurrent glioblastomas. Stromelysin was characteristically not expressed in any gliomas, and the expression level of TIMP-2 did not significantly change in the gliomas. These results suggest that the concomitant increased expression of gelatinase A, gelatinase B and occasional matrilysin genes is associated with the malignancy of gliomas and accompanied by the increased expression of TIMP-1 gene.

Astrocytoma

DNA image cytometry and the expression of proliferative markers (proliferating cell nuclear antigen and Ki67) in non-Hodgkin's lymphomas.

We have analyzed DNA content and proliferative activity in morphologically defined cell subpopulations of 74 non-Hodgkin's lymphomas (NHL) and 29 reactive lymph nodes using DNA image cytometry and antibodies to proliferative markers (proliferating cell nuclear antigen (PCNA) and Ki67). Thirteen (18.6%) of 70 NHL cases were aneuploid. The follicular center cell-derived lymphomas with DNA aneuploidy had DNA indices (DI) predominantly in the tetraploid region, whereas aneuploid high-grade (HG) NHL presented DNA histograms with multiple aneuploid stemlines. In aneuploid centrocytic-centroblastic (CB/CC) NHLs, DNA aneuploidy was found exclusively in centroblasts, whereas centrocytes in these cases were diploid. Percentages of cells in S and G2/M phase in chronic lymphocytic leukemia (CLL), immunocytoma (IC), centrocytic NHL (CC), and centrocytes from CB/CC were low (< 5%), whereas the respective values for centroblasts in CB/CC and in malignant cells of HG NHL were similar to those of large lymphoid cells in the reactive lymph nodes (mean, 39.5%, 36.6%, and 53.5%, respectively). The mean percentage of PCNA positive cells in CLL, IC, and CC was 4.9%. In the follicles of CB/CC NHLs there was, on average, 56.9% of PCNA positive centroblasts and 8.1% of PCNA positive centrocytes. In HG NHL, the mean percentage of PCNA positive lymphoma cells was 27.9%. A positive correlation was found between percentages of cells in S and G2/M phase and cells positive for PCNA (P < 0.001). There was also a significant correlation between percentages of Ki67 (mean, 19.2%) and PCNA positive cells (mean, 17.7%) (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

The immunohistochemical distribution of protein kinase C isozymes is altered in the canine brain and basilar artery after subarachnoid hemorrhage.

The effects of subarachnoid hemorrhage on immunoreactivities of protein kinase C isozymes (alpha, beta, gamma) were studied in canine brain and basilar artery. Mild and severe constriction of the basilar artery was shown 2 days and 7 days after hemorrhage, respectively. In control brain tissues, the isozymes showed distinct distributions and following hemorrhage, reactive astrocytes with protein kinase C alpha staining emerged in the surface of the pons and hippocampus. The control basilar artery showed protein kinase C alpha staining and its staining was decreased on day 7, consistent with the result of immunoblot. Our results demonstrated that subarachnoid hemorrhage induces gliosis with heavy protein kinase C alpha staining and down-regulation of protein kinase C in the artery.

Animals

Stimulation of protein-tyrosine phosphorylation in gerbil hippocampus after global forebrain ischemia.

Tyrosine phosphorylation in the gerbil hippocampus after a transient ischemia was analyzed by immunoblotting and immunohistochemistry. In control hippocampus, the phosphotyrosine was detected in many proteins of 165 to 10 kDa and the immunostain showed a distinct distribution. The ischemic insult induced various alterations of the phosphotyrosine immunoreactivities in both ischemia-resistant and -vulnerable neurons which were associated with alterations in the expression of 165 to 19 kDa-immunoreactive bands. These results suggest that tyrosine phosphorylation is involved in the ischemic hippocampus to play a role in the development of early and delayed neuronal deaths in CA4 and CA1 neurons, respectively.

Animals

Immunohistochemical differential diagnosis of benign cysts in the central nervous system.

This report concerns the immunohistochemical characterization of 6 cases of thin-walled cysts in the central nervous system (enterogenous cyst, paraphyseal neuroepithelial cyst, Rathke's cleft cyst and arachnoid cyst). Antibodies to glial fibrillary acidic protein (GFAP), S-100 protein, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), keratin (KER) and vimentin (VIM) were used. The enterogenous cyst was positive for KER, EMA and CEA. The neuroepithelial cyst of paraphyseal origin was positive for KER and S-100. The three Rathke's cleft cysts were positive for KER and EMA, but negative for S-100 and CEA, and the arachnoid cyst was positive for EMA and VIM. A unified concept and classification of the types of cysts studied based on immunohistochemical assays are proposed.

Adolescent

Measurement of interferon sensitivity in tumour cells from fine-needle aspirations.

Previous studies have shown significant correlations between interferon (IFN) induced enhancement of the enzyme 2',5'-oligoadenylate (2',5'-A) synthetase in vitro and response to IFN therapy. A limitation of this and other predictive tests is the availability of malignant cells for culture. Malignant cells can be obtained from most palpable solid tumours by fine-needle aspiration. We investigated whether malignant cells from such aspirations can be used in a 2',5'-A synthetase assay. In 23/27 (85%) of the cases sufficient amounts of viable cells were obtained, containing a high proportion (greater than or equal to 90%) of tumour cells. In 13/23 tumour samples (57%) IFN-alpha significantly enhanced the 2',5'-A synthetase levels. The use of cells from the fine-needle aspirations for prediction of IFN sensitivity, makes the 2',5'-A synthetase test applicable in a wide range of tumours at a variety of disease stages.

2',5'-Oligoadenylate Synthetase

Fine needle aspiration cytology of high grade T-cell lymphomas in human T-lymphotropic virus type 1 carriers.

We studied the fine needle aspiration (FNA) cytology of high grade peripheral T-cell lymphomas from eight human T-lymphotropic virus-1 (HTLV-1) positive patients. FNA smears from seven lymphomas showed a distinctive cytologic pattern with a dominance of rounded cells with irregular nuclei and a moderately basophilic cytoplasm. Irregular cells with a pale abundant cytoplasm were present in varying amounts. Some smears contained a few giant cells with cerebriform nuclei. In addition, plasma cells and eosinophils were found. Epithelioid cells were an inconstant finding. On histology these seven lymphomas were assigned to the pleomorphic medium-large cell subtype and all but one were of T-helper phenotype with rearrangements of the T-cell receptor. FNA smears from a lymph node in a patient with a previous histological diagnosis of lymphomatoid papulosis of the gingiva showed a monotonous pattern of large immunoblastic cells with some binucleated variants consistent with a diagnosis of high grade immunoblastic lymphoma, which was confirmed histologically. Our results show that peripheral T-cell lymphomas from HTLV-1 positive patients have cytological patterns which are distinctive enough to allow a conclusive diagnosis of high grade T-cell lymphoma. However, we do not think that the cytology of HTLV-1 positive lymphomas can be differentiated from that of virus-unrelated high grade T-cell lymphomas.

Adult

Endothelin-1 of canine basilar artery in vasospasm.

Cerebral vasospasm was induced in adult mongrel dogs by a two-hemorrhage method. The basilar arteries were quickly frozen after careful removal of surrounding blood clot and their level of immunoreactive endothelin-1, a strong vasoconstrictor produced by the endothelial and vascular smooth-muscle cells, was measured by sandwich-enzyme immunoassay. The levels of immunoreactive endothelin-1 (mean +/- standard deviation) were 112.9 +/- 7.0 pg/mg protein prior to vasospasm, 180.4 +/- 24.7 pg/mg protein on Day 2 after vasospasm, and 115.0 +/- 24.0 pg/mg protein on Day 7, showing a significant increase (p less than 0.01) in immunoreactive endothelin-1 only on Day 2. In addition, vasospasm was moderately reversed by the topical application of monoclonal antibody against endothelin-1 on Day 2 but rather resistant to topical monoclonal antibody on Day 7. It is suggested that endothelin-1 could act as a trigger in the early stages of cerebral vasospasm, but that the maintenance of cerebral vasospasm at later stages might be independent of endothelin-1.

Animals

Effects of inhibitors of protein kinase C and calpain in experimental delayed cerebral vasospasm.

Vasospasm was produced in adult mongrel dogs by a two-hemorrhage method, and the spastic basilar arteries were exposed via the transclival route on Day 7. Tonic contraction was produced in the normal canine basilar arteries by a local application of KCl or serotonin after transclival exposure. The exposed spastic and tonic basilar arteries then received a topical application of the following: 1-(5-isoquinolinesulfonyl)-2-methyl-piperazine (H-7), a potent inhibitor of protein kinase C acting at the catalytic domain; calphostin C, a specific inhibitor of protein kinase C acting at the regulatory domain; or calpeptin, a selective inhibitor of calpain. Both spastic and tonic basilar arteries were effectively dilated by H-7. Calphostin C caused only slight dilation of spastic basilar arteries but moderate dilation of tonic basilar arteries. Dilation in response to calpeptin was remarkable in the spastic basilar arteries but slight in the tonic basilar arteries. The doses of calphostin C and calpeptin required to obtain maximum effect were markedly lower in the tonic model than in the spastic model. The spastic and tonic models had a similar dose-dependent response to H-7 but quite a different response to calphostin C or calpeptin, suggesting a difference in the function of protein kinase C and calpain in the two models. Furthermore, the effect of calphostin C on the reversal of vasospasm was increased significantly after topical treatment with calpeptin. It is suggested that the majority of the catalytic domain of protein kinase C is dissociated from the regulatory domain, probably by a limited proteolysis with calpain, and is markedly activated in vasospasm.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine