Biomedical subjects

H Saitoh

Publications and source records attributed to H Saitoh.

At least 73 records · Page 4Linked to original sources

PubMed2000

Expression of tumor necrosis factor-alpha--converting enzyme and tumor necrosis factor-alpha in human myocarditis.

OBJECTIVES: We determined whether tumor necrosis factor-alpha-converting enzyme (TACE) is expressed with tumor necrosis factor-alpha (TNF-alpha) in myocarditis. BACKGROUND: Tumor necrosis factor-alpha-converting enzyme, which has recently been identified as belonging to the family of metalloproteinase disintegrin proteins, is responsible for the conversion of TNF-alpha precursor to its mature form. METHODS: We examined TACE and TNF-alpha expressions in endomyocardial biopsy tissues obtained from 14 patients with myocarditis and five control subjects by using quantitative reverse transcriptase polymerase chain reaction and immunohistochemistry. RESULTS: Expression of TNF-alpha and TACE messenger ribonucleic acid (mRNA) was significantly greater in the myocarditis group than in the control group. A positive correlation was found between TNF-alpha and TACE mRNAs (r = 0.83, p < 0.05). Six patients with severe myocarditis underwent repeat biopsies. Although TNF-alpha and TACE mRNAs were expressed at high levels in the initial biopsies, a marked decrease was noted in the repeat biopsies. The immunostainings for TNF-alpha and TACE were positive in the myocytes and interstitial cells of myocardium obtained from patients with myocarditis. Expression of TACE and TNF-alpha mRNAs was greater in the subgroup in New York Heart Association functional class III or IV than in the subgroup in class I or II. Expression of TACE and TNF-alpha mRNA was correlated positively with left ventricular volume (TNF-alpha: r = 0.85; TACE: r = 0.80) and negatively with left ventricular systolic function (TNF-alpha: r = -0.85; TACE: r = -0.85). CONCLUSIONS: These findings indicate that the expression of TNF-alpha and TACE may have important implications in the pathogenesis of myocarditis and may influence advanced cardiac dysfunction in myocarditis.

ADAM Proteins↗

PubMed2000

Region-dependent disappearance of vinblastine in rat small intestine and characterization of its P-glycoprotein-mediated efflux system.

This study was aimed to characterize the absorption behavior of vinblastine (VLB), a well-known substrate of P-glycoprotein (P-gp), from rat small intestine, especially focusing on the regional-dependence of its efflux mediated by P-gp. VLB disappeared from duodenal and ileal loops of male Wistar rats fairly rapidly (30-60% in 30 min). In contrast, its disappearance from the jejunal loop was almost negligible and in some rats >100% of the jejunal dose was recovered. The radioactivity derived from [3H]VLB, which was absorbed from duodenum and ileum, was detected in the jejunal region. The jejunal appearance of radioactivity was increased when unlabeled VLB was present in the region in advance. The basolateral-to-apical transport of [3H]VLB across Caco-2 cell monolayers was greater when unlabeled VLB was added to the apical medium than when VLB-free buffer was applied to the apical side. When verapamil or cyclosporin A, potent modulators of P-gp, was added to the apical medium together with unlabeled VLB, enhanced basolateral-to-apical transport of [3H]VLB was disappeared. It is suggested that VLB absorption is strongly restricted by P-gp, especially in the jejunal region of the rat small intestine, and that the secretory transport via intestinal P-gp may be subject to trans-stimulation. Moreover, intravenously administered methylprednisolone and intramuscularly administered progesterone significantly enhanced the absorption of VLB, suggesting that parenterally administered P-gp modulators could influence the intestinal absorption of P-gp substrates.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2000

Larval susceptibility of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae), to Bacillus thuringiensis H serovars isolated in Japan.

A total of 1700 Japanese strains of Bacillus thuringiensis, belonging to at least 47 H serogroups, were examined for insecticidal activity against larvae of the diamondback moth, Plutella xylostella. The high-level toxicity was associated with 612 isolates (36.0%). Of these, 608 isolates (99.3%) fell into 13 H serogroups belonging to the low-numbered H serotypes, H1-H10. Conversely, most isolates belonging to the high-numbered serotypes (>H10) had little or no larvicidal activity; only one isolate of the serovar japonensis H23 was active. P xylostella larvae were susceptible to 89.8% of the serovar morrisoni H8a:8b strains and 85.7% of galleriae H5a:5b strains. High values of 60-80% were also obtained in six serovars (thuringiensis H1, alesti H3a:3c, kurstaki H3a:3b:3c, kenyae H4a:4c, aizawai H7, and tolworhi H9), while relatively low values of <60% in two other common serovars, sotto H4a:4b and darmstadiensis H10a:10b. Five selected isolates, belonging to H serovars other than kurstaki and aizawai, were 10-60 times less toxic than the reference strain HD-1 (serovar kurstaki). Parasporal inclusion proteins of these strains were immunologically unrelated to those of the strain HD-1 and the aizawai type strain.

Animals↗

PubMed2000

Nerve growth factor prevents apoptosis of cord blood-derived human cultured mast cells synergistically with stem cell factor.

BACKGROUND: Stem cell factor (SCF) has been identified as a critical survival factor of human mast cells. Other cytokines which possess survival promotion activity on human mast cells are less known. OBJECTIVE: We examined the survival promotion activity of nerve growth factor (NGF) on cord blood-derived human cultured mast cells. METHODS: Expression and function of NGF receptors on the mast cells were examined by RT PCR, flowcytometric analysis, immunoprecipitaion and western blotting. The survival promotion activity of NGF to the mast cells was examined. To evaluate the proliferating activity of NGF on the human cultured mast cells, flow cytometric analysis with propidium iodide staining was applied. To confirm whether the human mast cell growth activity of NGF was caused by a suppression of apoptosis, the proportion of the cells containing in situ DNA fragmentation was counted. RESULTS: The human cultured mast cells expressed the high affinity receptor p140trk but not the low affinity receptor p75LNGFR. NGF induced the phosphorylation of p140trk. NGF alone could not support the survival of the mast cells, however, the addition of NGF to the culture medium containing recombinant SCF led to a significant increase of the number of survival mast cells. No significant changes of the cell cycle from G0/G1 phase to the S/G2 + M phases were observed by NGF. In contrast, the addition of NGF to the medium with SCF showed a significant inhibitory effect on the apoptosis of the mast cells. CONCLUSION: NGF may act as a key factor to promote the survival of human mast cells synergistically with SCF through the prevention of apoptosis.

Apoptosis↗

PubMed2000

Serum levels of free insulin-like growth factor (IGF)-I and IGF-binding protein-1 in prepubertal children with idiopathic short stature.

The study was performed to evaluate the relationships among serum free and total insulin-like growth factor (IGF)-I, IGF-binding protein-1 (IGFBP-1), IGFBP-3, and insulin concentrations in prepubertal children with idiopathic short stature (ISS). Eighteen children with ISS and 15 age-matched controls were included in the study. All short children had a height standard deviation score of more than 2 below the mean, and maximum stimulated GH levels greater than 10 microg/l after two standard provocation tests. The serum levels of free IGF-I were significantly lower in short children (1.6 +/- 0.3 microg/l) than in the controls (2.8 +/- 0.6 microg/l, P<0.05), while total IGF-I levels were slightly, but not significantly, lower in short children than in controls. The serum levels of IGFBP-1 were significantly higher in the ISS group (124.6 +/- 5.6 microg/l) than in controls (80.0 +/- 8.7 microg/l, P < 0.0001). The fasting insulin and IGFBP-3 levels were similar in both groups. A stepwise regression analysis for all subjects revealed that IGFBP-1 is the only independent predictor of log free IGF-I (R2 = 0.23, P<0.01). The present study shows that the serum levels of free IGF-1 are significantly lower and insulin-like growth factor-binding protein-1 levels are higher in prepubertal children with idiopathic short stature, as compared with age-matched controls. The high IGFBP-1 may contribute to growth retardation in a subgroup of idiopathic short stature through a decrease in free IGF-1.

Case-Control Studies↗

PubMed2000

In vitro cytotoxicity of non-cyt inclusion proteins of a Bacillus thuringiensis isolate against human cells, including cancer cells.

A soil isolate designated 90-F-45-14, belonging to Bacillus thuringiensis serovar dakota (H15), was examined for characterization of in vitro cytotoxicity, associated with parasporal inclusion proteins, against human cells. When activated with proteolytic processing, inclusion proteins of the isolate 90-F-45-14 exhibited a moderate cytotoxicity against the human uterus cervix cancer cells (HeLa) with an EC(50) value of 60.8 microg ml(-1), while showing extremely high activities on the human leukaemic T cells (MOLT-4) and the normal T cells with EC(50) values of 0.27 and 0.20 microg ml(-1), respectively. Anti-leukaemic cell activity of the 90-F-45-14 proteins was eight to nine times greater than that of the B. thuringiensis serovar israelensis proteins containing the Cyt1 protein, a broad-spectrum cytolysin. The cytopathy by the 90-F-45-14 proteins was characterized by marked cell-ballooning, while the israelensis proteins induced early breakdown of the cells due to cytolysis. Inclusions of the isolate consisted of five major polypeptides of 170, 103, 73, 40 and 32 kDa. A 100% homology was observed in the sequence of 15 N-terminal amino acids between the proteins of 170 and 103 kDa. There was no N-terminal sequence homology between 90-F-45-14 proteins and the existing Cry/Cyt proteins of B. thuringiensis. Proteolytic processing by proteinase K yielded several proteins with molecular masses ranging from 40 to 28 kDa.

Amino Acid Sequence↗

PubMed2000

Pharmacokinetics of landiolol hydrochloride, a new ultra-short-acting beta-blocker, in patients with cardiac arrhythmias.

OBJECTIVES: To elucidate pharmacokinetics and pharmacodynamics of landiolol hydrochloride, newer developed ultra-short-acting beta-blocker, in patients with various cardiac tachyarrhythmias. BACKGROUND: The short duration of action and titratability of landiolol hydrochloride make it ideal for use in patients with a clinical need for beta-blockers. METHODS: In a total of 31 examinations we infused the drug in 19 patients (mean age, 55 +/- 14 years). After the persistence of the tachyarrhythmias was confirmed, continuous infusion was started at rates of 0.005, 0.01, 0.02, 0.04, and 0.08 mg/kg/min for 5 minutes (for paroxysmal atrial fibrillation, paroxysmal supraventricular tachycardia, and ventricular tachycardia) or 15 minutes (for ventricular premature complex). We analyzed the pharmacokinetics of 16 examinations. A one-compartment model provided a close fit for each blood concentration-time curve. RESULTS: The maximum blood concentrations obtained clearly showed the dose dependency and revealed very short half-lives (range, 2.3 to 4.0 minutes). Area under the blood concentration-time curves also increased, showing dose dependency. In paroxysmal atrial fibrillation, landiolol hydrochloride reduced the heart rate from 111 +/- 20 to 90 +/- 10/min. Sinus rhythm was restored, without any adverse effects, in three of five patients with paroxysmal supraventricular tachycardia and one patient with ventricular tachycardia. There was no significant change in peripheral blood pressure. CONCLUSIONS: Landiolol hydrochloride has a shorter elimination half-life than any other beta-blocker, and it can be administered safely to patients with various tachyarrhythmias.

Adolescent↗

PubMed2000

Parasporin, a human leukemic cell-recognizing parasporal protein of Bacillus thuringiensis.

An unusual property, human leukemic cell-recognizing activity, associated with parasporal inclusions of a noninsecticidal Bacillus thuringiensis soil isolate was investigated, and a protein (named parasporin in this study) responsible for the activity was cloned. The parasporin, encoded by a gene 2,169 bp long, was a polypeptide of 723 amino acid residues with a predicted molecular weight of 81, 045. The sequence of parasporin contained the five conserved blocks commonly found in B. thuringiensis Cry proteins; however, only very low homologies (<25%) between parasporin and the existing classes of Cry and Cyt proteins were detected. Parasporin exhibited cytocidal activity only when degraded by proteases into smaller molecules of 40 to 60 kDa. Trypsin and proteinase K activated parasporin, while chymotrypsin did not. The activated parasporin showed strong cytocidal activity against human leukemic T cells (MOLT-4) and human uterus cervix cancer cells (HeLa) but not against normal T cells.

Amino Acid Sequence↗

PubMed2000

Antisense oligonucleotides to NF-kappaB improve survival in bleomycin-induced pneumopathy of the mouse.

We examined the effect of antisense oligonucleotides to the p65 subunit of NF-kappaB on the survival of bleomycin (BLM)-induced pneumonitis in C57BL/6 mice (female, 8 wk of age, 17 to 20 g body weight). Fifty-three percent and all control mice died within 6 to 9 d after intravenous administration of 150 and 300 mg/kg BLM alone, respectively. The intravenous administration of the antisense oligonucleotides (900 microgram/animal dissolved in 200 microliter saline, 6 h before and 5 d after BLM administration) significantly improved the survival rate to 100 and 40% in 150- and 300-mg/kg-treated animals, respectively. The antisense oligonucleotides also significantly improved the loss of body weight, the increase in lung hydroxyproline, and histologic changes by BLM, whereas the antisense oligonucleotides themselves did not produce any significant changes in the behavior or lung histology. Both peripheral blood monocytes and alveolar macrophages were confirmed to contain large amounts of intracellular antisense oligonucleotides after BLM injection by FITC-labeled fluorescent microscopy. Further, Western blotting confirmed the inhibition of NF-kappaB in macrophages by the antisense oligonucleotides. These findings suggest that the antisense oligonucleotides are incorporated into activated alveolar macrophages and ameleriorate the lung injury and pneumonitis/fibrosis, thereby improving the survival of BLM-induced pneumopathy in mice.

Animals↗

PubMed2000

A novel function of thyrotropin as a potentiator of electrolyte secretion from the tracheal gland.

Accumulating evidence suggests that thyrotropin (thyroid-stimulating hormone [TSH]) plays some roles in immunoregulation by an extrathyroidal action. Because airway submucosal glands are responsible for nonspecific and specific airway defense, we tested the effect of TSH on feline tracheal submucosal gland using a whole-cell patch-clamp technique, immunohistochemistry, and reverse transcription/polymerase chain reaction (RT-PCR). TSH potentiated neurotransmitter-induced ionic currents significantly in a dose-dependent manner. Acetylcholine (10(-)(8) M)- and norepinephrine (10(-)(7) M)-induced inward current (I(i)), which we previously showed to be a Cl(-) current, were increased to about 3-fold the pre-TSH control responses, respectively, by 2.0 ng/ml TSH; and to 6- and 23-fold the control values by 20.0 ng/ml TSH, respectively. TSH alone was without effect up to 20.0 ng/ml. Follicular stimulating hormone only slightly affected the I(i) (1. 5-fold the control). Analyses with immunohistochemistry and RT-PCR failed to identify TSH receptors on the glandular tissue. Maneuvers to raise the cellular adenosine 3',5'-cyclic monophosphate also failed to mimic the TSH-mediated potentiation. The TSH effect appeared to be mediated by a signaling pathway involving tyrosine kinase because its inhibitors (genistein and herbimycin A) abolished the augmentation completely, and interferon-gamma, a tyrosine kinase activator, imitated the TSH action on submucosal gland. Thus, TSH may be an important regulator of airway fluid secretion.

1-Methyl-3-isobutylxanthine↗

PubMed2000

[Toxoplasma pericarditis without immunosuppressant disorder detected by polymerase chain reaction of pericardial fluid: a case report].

There have been several case reports, a total of 22 up to the present, of toxoplasma pericarditis. Out of them, in only a few cases the diagnosis was properly made with a proof of the microscopic presence of Toxoplasma gondii. This is the first report of toxoplasma pericarditis in which the presence of Toxoplasma gondii was detected by polymerase chain reaction of pericardial effusion. In addition, the previous reports will be reviewed, and compared to this present case. A 29-year-old woman, without immunosuppressant disorder, suffering from fever and orthopnea was admitted to our hospital. Blood chemistry findings indicated mild liver dysfunction and inflammation. Chest radiography showed cardiac enlargement. Electrocardiography showed sinus tachycardia and ST elevation. Echocardiography revealed a massive pericardial effusion. Pericardiocentesis demonstrated 638 ml of bloody fluid. Cytologic study of the fluid was class II for malignancy, and polymerase chain reaction to tuberculosis was negative. However, a high titer of the anti-toxoplasma antibody of 1: 20,480 (passive hemagglutination) indicated pericarditis caused by Toxoplasma gondii. Subsequently, Toxoplasma gondii was identified in the pericardial effusion by polymerase chain reaction. Clinical symptoms improved after pericardiocentesis, but 2 months later pericarditis recurred. Treatment was started with 800 mg acetylspiramycin daily but failed to improve the symptoms. Because of the development of pleuritis, treatment was changed to sulfadoxine 1,000 mg/pyrimethamine 50 mg. After the treatment with them, her symptoms improved. Only 22 cases of toxoplasma pericarditis have been reported worldwide and 15 of those cases were without immunosuppressant disorder. The usual symptoms at the onset of pericarditis without immunosuppressant disorder are fever, dyspnea and chest pain. Seven patients developed cardiac tamponade. Pericardiocentesis was performed in 8 cases and the pericardial fluid was hemorrhagic in 6. Pericardial thickening was detected in 5 cases. The diagnosis of toxoplasma infection is very difficult, because asymptomatic infection of Toxoplasma gondii is very common. Pericarditis is a disease difficult to confirm the etiology. Detection of Toxoplasma gondii in pericardial effusion by the polymerase chain reaction is very useful for its diagnosis.

Adult↗

PubMed2000

[A case of epithelioid leiomyoma (leiomyoblastoma) of the urethra].

A 25-year-old woman was admitted to our hospital because of a painless, slow growing mass arising from the external genitalia. The mass had been developing for a few years. She did not have difficulty voiding nor was there hematuria. Magnetic resonance imaging revealed a well-circumscribed, 4 cm, solid tumor between the urethra and the anterior vaginal wall. After needle biopsy confirmed a benign tumor arising from the smooth muscle, the tumor was removed by a transvaginal approach. The histopathological diagnosis was epithelioid leiomyoma (leiomyoblastoma), which often occurs in the stomach or uterus, but seldom around the urethra. The patient has remained well without recurrence for 20 months after surgery.

Adult↗

PubMed2000

Therapy-related leukemia and myelodysplastic syndrome: a large-scale Japanese study of clinical and cytogenetic features as well as prognostic factors.

It is known that alkylating agents and topoisomerase II inhibitors can cause distinct forms of therapy-related leukemia and myelodysplastic syndrome (TRL/MDS). Although several reports have been made on each of these agents separately, no study has yet been conducted to evaluate the effect of these two types of agents in the same population. In a nationwide, large-scale population study, the clinical and cytogenetic features as well as the prognostic factors in 256 patients with TRL/MDS were assessed. Median age was 61 years, and the median period of latency from primary malignancies was 47.9 months. The latency period was significantly shorter in patients undergoing chemotherapy, especially that of topoisomerase II inhibitors, for primary cancer. The morphological diagnosis of TRL/MDS was acute myeloid leukemia in 59% and MDS in 41% of patients. Chromosome abnormalities that frequently involved chromosomes 5, 7 or 11 were documented in 77% of the 189 patients examined. MLL gene rearrangements were detected in 11 of 58 subjects and were correlated with a borderline significance (P = 0.072) with topoisomerase II inhibitor administration. Overall median survival was only 9.7 months. Survival was similar in cases with or without MLL gene rearrangement. Multivariate analysis identified chromosome 5 abnormalities, hypoproteinemia, poor therapy outcomes for primary cancer, C-reactive protein, and thrombocytopenia as being significantly poor prognostic factors (P < 0.05). This large-population study provided a comprehensive update of TRL/MDS status in Japan, identified significant prognostic factors, and enabled the clinical significance of MLL gene rearrangement to be assessed.

Adult↗

PubMed2000

[Crossed fused kidney with ectopic ureter: a case report].

A 5-year-old girl with a rare anomaly of a single system ectopic ureter draining into the mesonephric duct cyst and ipsilateral crossed fused kidney is presented. Both ureters were catheterized and three dimensional (3-D) images of the urinary tract were reconstructed with helical computed tomographic (CT) data. A mesonephric duct cyst, the right ureter draining into the cyst and an ectopic orifice were well depicted with these images, especially with inferior projection images. 3-D display of helical CT data with a retrograde contrast study is a promising method of obtaining anatomical detail of an ectopic ureter.

Child, Preschool↗

PubMed2000

[A case of giant hemorrhagic adrenal pseudocyst with infection].

A 51-year-old woman was admitted with general fatigue, high fever and left upper abdominal pain. Abdominal computed tomography revealed a left adrenal mass, 15 cm in diameter with regular margins, the contents of which were not enhanced with contrast medium. The mass had heterogeneous echogenicity on the abdominal ultrasonograph. Serum hormonal levels were almost normal. Despite treatment with antibiotics, the high fever persisted. Based on these findings, the differential diagnosis was adrenal abscess or a nonfunctional adrenal tumor. Transabdominal left adrenalectomy was performed. Pathological examination revealed a hemorrhagic adrenal pseudocyst with infection. The patient's condition improved soon after the operation.

Adrenal Gland Diseases↗

PubMed2000

[Comparative evaluation of BACTEC MGIT 960 system with MB/BacT and egg-based media for recovery of mycobacteria].

Fully automated, nonradiometric mycobacteria culture systems, BACTEC MGIT 960 (Becton Dickinson Microbiology Systems, Sparks, Md, U.S.A.) and MB/BacT (Organon Teknika, Durham, NC, U.S.A.) were evaluated in comparison with three different eggbased media (3% Ogawa, Ogawa K, and Vite) for the ability to detect mycobacteria in clinical sputum specimens. Sputum specimens were processed by semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) for the automated systems, and by cetylpyridium chloride-succinic acid-NaCl for the egg-based media. A total of 954 sputum specimens were processed, and the recovery of mycobacteria by the BACTEC MGIT 960 was performed in a commercial laboratory. Overall, the frequency of breakthrough contamination was <1% for the three egg-based media, ranging from 0. 42% to 0.63%. Whereas, the frequency of false positives due to breakthrough contamination was 1.89% for MB/BacT and 20.1% for BACTEC MGIT 960. A total of 237 isolates of Mycobacterium tuberculosis complex and 167 isolates of nontuberculous mycobacteria (NTM) were recovered. The highest recovery ratio was obtained by MB/BacT (95.8%), followed by the egg-based media, Vite (74.3%), Ogawa K (65.8%), and 3% Ogawa (58.9%). The recovery ratio by BACTEC MGIT 960 was the lowest, and estimated at 43.1%, mainly due to a high frequency of breakthrough contamination. However, even after omission of these false positives reported by BACTEC MGIT 960, the recovery ratio by this system was comparable to that of 3% Ogawa media. The time to detection of 50% of positive cultures of M. tuberculosis complex by BACTEC MGIT 960 and MB/BacT was 20 days and 17 days, respectively.>From these results, it may be concluded that MB/BacT is superior to BACTEC MGIT 960 and egg-based media for the recovery of mycobacteria from sputum specimens. Furthermore, based on the outcome of this study, we think that considerable improvements are necessary for the clinical application of BACTEC MGIT 960. These improvements should particularly be focused on reducing the false positive ratio caused by contamination, and culture media, which effectively support mycobacterial growth.

Bacteriological Techniques↗

PubMed2000

[Application of DNA-DNA hybridization, DDH MYCOBACTERIA 'Kyokuto' to species-identification of mycobacteria grown in Middlebrook 7H9 broth].

A colorimetric DNA-DNA hybridization for the genetic identification of mycobacteria, DDH MYCOBACTERIA 'KYOKUTO' (Kyokuto Pharmaceuticals, Tokyo) was evaluated for the clinical isolates of mycobacteria grown in Middlebrook 7H9 broth of MB/BacT (Organon Teknika, Durham, NC, U.S.A.). When the MB/BacT gave the positive interpretation, 10 ml of Middlebrook 7H9 broth was collected from the bottle. After centrifugation at 3,000 rpm for 10 min, two drops of acetone were added to the pellet, then let it stand for one hour at the room temperature. The air-dried pellet was resuspended in a small volume of distilled water, and proceeded to the identification described. Of 136 clinical isolates of mycobacteria, comprising of 76 M. tuberculosis complex and 60 nontuberculous mycobacteria (NTM), ninety-five (70%) were correctly identified when compared to the reference identification. Thirty (22%) isolates resulted in the unidentified due to negative reaction throughout the test wells, and the remaining 11 (8%) were also unidentified due to low likelihoods. According to the package insert, the DDH MYCOBACTERIA may not be applicable to the isolates grown in Middlebrook broth. However, our test procedure using acetone prior to the extraction of bacterial DNA enables us to directly identify the isolates of mycobacteria grown in Middlebrook 7H9 broth of MB/BacT.

Acetone↗

PubMed2000

Characterization of the anti-cancer-cell parasporal proteins of a Bacillus thuringiensis isolate.

An unusual activity, associated with non-insecticidal and non-haemolytic parasporal inclusion proteins of a Bacillus thuringiensis soil isolate, designated 89-T-26-17, was characterized. The parasporal inclusion of this isolate was bipyramidal, rounded at both ends, containing proteins of 180, 150, 120, 100, and 88 kDa. No homologies with the Cry and Cyt proteins of B. thuringiensis were detected based on N-terminal sequences. Proteolytic processing of the inclusion proteins by proteinase K, trypsin, and chymotrypsin produced a major protein of 64 kDa exhibiting cytocidal activity against human leukaemic T cells and uterus cervix cancer (HeLa) cells. The protease-activated proteins showed no cytotoxicity to normal T cells.

Amino Acid Sequence↗