Screening of endophytic fungi having ability for antioxidative and alpha-glucosidase inhibitor activities isolated from Taxus sumatrana.

Endophytic microbes are considered as an important source of natural products. They show antibiotic, anticancer, antioxidative and antidiabetic activities. Therefore, there are many reports on the isolation and bioactivity screening of endophytic fungi from various plants including Taxus species. Taxus sumatrana (Miq.) de Laub is found in Indonesia. The objective of this study is to conduct an in vitro screening of 14 endophytic fungi isolated from Taxus sumatrana having antioxidative and alpha-glucosidase inhibitor activities. Each endophytic fungus was cultured for 7 days and the fungal mycelium and medium were extracted with methanol and ethyl acetate, respectively, to produce each extract. The antioxidative activity of each extract was tested by DPPH free radical scavenging activity and beta-carotene bleaching assays, whereas antidiabetic activity was tested based on alpha-glucosidase inhibitor activity. The screening results showed that fungal mycelia of TSC 13 had the best alpha-glucosidase inhibitor activity and TSC 24 had the best antioxidative activity. Isolation of bioactive compounds from TSC 13 and TSC 24 is being conducted. This is the first report that endophytic fungi isolated from T. sumatrana exhibited anti alpha-glucosidase inhibitory and anti oxidative activities.

Screening of fungi capable of degrading lignocellulose from plantation forests.

In an effort to prevent forest fires after the clear cutting of plantation forests, fungi capable of degrading lignocelluloses were isolated to make a fertilizer from the logging waste. Seventy five fungal species were isolated from fruiting bodies and mycelia in plantation forests of South and North Sumatera, Indonesia. Sixty three of the fungi were identified based on the appearance and morphological characteristics of their fruiting bodies and mycelia, as Pycnoporus sanguineus, Dacryopinax spathularia, Schizophyllum commune, Polyporus sp. and Trametes sp. Twenty fungi were categorized as white-rot fungi and 12 as brown-rot fungi. Moreover, isolates 371, 368, 265, 346, 345 and 338 were selected using indicators and tested for the ability to degrade lignin and holo-cellulose in mangium wood meal over 1 to 4 weeks. Results showed that the 6 fungi could degrade lignin and holo-cellulose in wood meal. An increase in incubation time tended to decrease the amounts of lignin and holo-cellulose. Isolate 371 was found to be best at degrading lignin and holo-cellulose in mangium wood meal.

Nocistatin attenuated the nociceptin induced c-Fos expression in the mouse hippocampus.

Nocistatin and nociceptin/orphaninFQ (N/OFQ) are the two new peptides which may have roles in nociception, memory, anxiety, and other biological functions. Nocistatin acts as a functional antagonist to N/OFQ in several functions, but their neuro-anatomical sites of interaction are unknown. We investigated the effect of combined intracerebroventricular (i.c.v.) injection of nocistatin with N/OFQ, on N/OFQ induced c-Fos expression in the mouse hippocampus, using c-Fos immunohistochemistry. We found that co-injection of nocistatin with N/OFQ significantly attenuated N/OFQ induced c-Fos expression in the hippocampus.

Combination chemotherapy with docetaxel and nedaplatin for recurrent esophageal cancer in an outpatient setting.

The purpose of this study was to address the feasibility of combination chemotherapy of docetaxel and nedaplatin for recurrent esophageal cancer patients in an outpatient setting. Patients received docetaxel (30 mg/m(2) intravenously) on day 1 and nedaplatin (40 mg/m(2) intravenously) on day 1 every 2 weeks. In total, 28 patients with recurrent esophageal cancer after the initial treatment (esophagectomy, chemotherapy and/or chemoradiotherapy) were enrolled. Each patient received six cycles of treatment and was evaluated with a computed tomography scan. The percentage of patients who completed this therapy was 60.7%. Complete response and partial response were achieved by 3.6% and 35.7% of patients, respectively. The most frequent toxicities were leukopenia and anemia; non-hematological toxicities were generally mild. There was no treatment-related death. The median survival time and 1-year survival rate were 8.5 months and 15.9%, respectively. This outpatient combination chemotherapy was useful as second-line chemotherapy for recurrent esophageal cancer.

Ganglion-associated ulnar tunnel syndrome treated by ultrasonographically assisted aspiration and splinting.

We describe a case of ulnar nerve compression at the wrist due to a ganglion. This was treated by aspiration of the ganglion under ultrasonography and splinting because the patient was pregnant. The ulnar nerve palsy resolved completely and the ganglion disappeared. A follow-up ultrasonographic examination after 2 years showed no recurrence of the ganglion.

Clinical significance of CD151 gene expression in non-small cell lung cancer.

Transmembrane 4 superfamily (TM4SF) is a recently described gene family, and TM4SF members are known to play roles in the signal transduction pathways and to regulate cell activation, development, proliferation, and motility. MRP-1/CD9, KAI1/CD82, and ME491/CD63, members of the TM4SF, have been reported to suppress tumor progression or metastasis. Previously, we showed that MRP-1/CD9 suppressed cell motility and metastatic potential to lungs. Moreover, reduction of MRP-1/CD9 and KAI1/CD82 gene expression was found to be a factor in a poor prognosis for patients with non-small cell lung cancer. However, among TM4SF, CD151 is identical to an existing gene, PETA-3, which may promote tumor metastasis of malignant cells, and its expression may be involved in the malignant progression of cancer. The function of CD151 is opposite that of the metastasis suppressor genes, MRP-1/CD9 and KAI1/CD82. On the basis of these results, we used reverse transcription-PCR and immunohistochemical techniques for a retrospective study of CD151 gene expression in tumor tissues from 145 lung cancer patients; 72 tumors were stage I, 29 stage II, 27 stage IIIA, and 17 stage IIIB. Whereas 86 patients had tumors positive for the CD151 gene, 59 had tumors that were negative for the CD151 gene. The overall survival rate of patients with CD151-positive tumors was much lower than that of CD151-negative patients (51.9% versus 73.1%; P = 0.013). Our findings suggest that high CD151 gene expression in lung cancer may be associated with a poor prognosis. Assessment of CD151 could be instrumental for improvements in lung cancer diagnosis and therapies.

Usefulness of tc-99m MIBI SPECT in predicting multidrug resistance gene expression levels in non-small cell lung cancer--a preliminary report.

In this study we investigated whether Tc-99m hexakis 2-methoxy isobutyl isonitnile (Tc-99m MIBI) single-photon emission computed tomography (SPECT) has a correlation with the multidrug resistance (MDRI) and multidrug resistance-associated protein (MRP1) gene expression levels in non-small cell lung cancer (NSCLC). Fifteen patients with NSCLC were studied. SPECT images were obtained 15 (early) and 120 (delayed) min after injection of Tc-99m MIBI. We chose only one transverse section and set the region of interest over the tumor and out of the body. The mean counts in the tumor on early and delayed images were corrected by using those in the background and represented as Te and Td, respectively. Resected tumor specimens were frozen with liquid nitrogen and each positive control cell line was cultured. After the total ribonucleic acid (RNA) was extracted from specimens and cell lines, the complimentary deoxyribonucleic acid (cDNA) was amplified by the reverse transcription-polymerase chain reaction (RT-PCR) method. Each product was electrophoresed and fluorointensity was measured. The gene expression level was represented as the ratio of that of the positive control cell line. Te and Td indicated a significant correlation with the MDR1 gene expression level (p = 0.015 and p = 0.022), but not the gene of MRP1 (p = 0.100 and p = 0.145). In conclusion, Te and Td are useful parameters in predicting the MDRI gene expression level, but not MRPI in NSCLC.

One-portal technique of endoscopic fasciotomy: Chronic compartment syndrome of the lower leg.

Many athletes complain of exercise-induced pain in the lower leg that can be caused by inflammatory diseases, peripheral nervous system disease, fatigue fracture, shin splint, and chronic compartment syndrome (CCS). CCS is the most typical exercise-induced condition and it often requires surgical decompression of the several compartments. There are already many techniques reported in the literature. Recently, an endoscopic technique for CCS was reported with which excellent results were achieved. We have modified it and developed a new technique for treating CCS of the lower leg. We report a case of CCS of the lower leg treated with 1-portal endoscopic fasciotomy. The technique helps to decrease damage to soft tissue and patients will immediately return to normal activities of daily living.

Neutral endopeptidase/CD10 and aminopeptidase N/CD13 gene expression as a prognostic factor in non-small cell lung cancer.

OBJECTIVES: Neutral endopeptidase modulates the growth of lung cancer, while aminopeptidase N degrades the extracellular matrix and is involved in cell motility. We studied the metastasis mechanism to detect novel metastasis-associated molecules and to evaluate them for clinical application. METHODS: We studied the relationship between the expression of neutral endopeptidase and aminopeptidase N by quantitative reverse transcript-polymerase chain reaction analysis in 132 patients with non-small cell lung cancer undergoing radical surgery from 1991 to 1996. RESULTS: Patients with neutral endopeptidase-positive and aminopeptidase N-negative tumors were defined as group A, those with neutral endopeptidase-positive and aminopeptidase N-positive or neutral endopeptidase-negative and aminopeptidase N-negative tumors as group B, and those with neutral endopeptidase-negative and aminopeptidase N-positive tumors as group C. The 5-year survival of group A patients (92.9%) was significantly better than that of group B patients (64.7%) and much better than that of group C patients (38.2%) (P = 0.0011). Neutral endopeptidase and aminopeptidase N thus have statistically significant P in overall survival in Cox regression (P = 0.019). CONCLUSION: Neutral endopeptidase and aminopeptidase N gene expressions are significant indicators of prognosis.

Hepatocellular carcinoma with a sarcomatous appearance: report of a case.

A 59-year-old man was admitted with general fatigue, an epigastric mass, and remittent fever. Radiological examinations disclosed a huge solid-to-cystic mass in the right lobe of the liver, and the mass severely compressed the right diaphragm, the inferior vena cava, and the right atrium. In addition, the patient suffered from chronic hepatitis; however, the serum alpha-fetoprotein, carcinoembryonic antigen, and PIVKA II levels were all within the normal ranges. The serum C-reactive protein level was 7.71mg/dl. With a clinical diagnosis of a malignant hepatic tumor invading the right diaphragm, surgery was performed. The tumor originated from segments IV and VII of the liver, was well defined, and grew extrahepatically. The tumor was resected using an ultrasonic cavitational aspirator together with the infiltrated right diaphragm. The resected tumor measured 23 x 13 x 23cm in size and weighed 3,700 g. Histologically, the tumor was found to consist of hepatocellular carcinomatous component and sarcomatous component. In the sarcomatous component, spindle-shaped cells which were positive for the immunohistochemical localization of vimentin, alpha-smooth muscle actin, and keratin were identified. The postoperative course was uneventful. The value of the serum C-reactive protein returned to within the normal range, and the patient became afebrile. The patient received a postoperative combination chemotherapy (etoposide, epirubicin, and cisplatin), and remains well with no signs of recurrence 12 months after the operation.

The use of ultrasound for drug delivery.

Ultrasound has been in use for the last three decades as a modality for diagnostic imaging in medicine. Recent studies have shown that nonthermal ultrasound energy could be applied for targeting or controlling drug release. This new concept of therapeutic ultrasound combined with drugs has induced interest in various medical fields. Enhanced effects of thrombolytic agents such as urokinase and TPA with acoustic energy have been demonstrated. Ultrasound transducer-tipped catheters are being developed for treatment of cardiovascular diseases. Other devices with ultrasound transducers implanted in transdermal drug patches are also being evaluated for possible delivery of insulin through the skin. Echo contrast microbubbles could also be used to carry and release genes to various tissues and lesions. Chemical activation of drugs by ultrasound energy for treatment of cancers is another new field recently termed "sonodynamic therapy." Various examples of ultrasound application are under investigation that could lead to revolutionary drug delivery systems of the future.

Spinal tanycytic ependymomas.

Three cases of spinal tanycytic ependymoma are reported, a man aged 45 years and two women aged 36 and 55 years. Each patient developed gradual paraparesis over a few months prior to admission. Magnetic resonance imaging showed an enhancing, well-circumscribed tumor in the spinal cord in each case. Histologically, the tumors consisted of monotonous proliferation of long spindle cells with markedly eosinophilic cell processes; focally forming perivascular pseudorosettes. The tumor cells were strongly immunopositive for glial fibrillary acidic protein, S-100 protein and vimentin. Ultrastructurally, in addition to massive intermediate filaments, many tumor cells showed abundant microtubules. Well-developed desmosomes and microvilli/cilia-lined microlumina were occasionally observed. The tumors were grossly totally removed and the patients remain recurrence free at 9, 9, and 2 years postoperatively. Reviewing reported cases including our three cases, tanycytic ependymoma may occur frequently in spinal cord, especially in the cervical region of the spinal cord. Since histologically it resembles pilocytic astrocytoma and schwannoma, tanycytic ependymoma should be included in the differential diagnosis of benign spindle cell tumors of the central nervous system.

A case report of early idiopathic portal hypertension.

We report herein a case (46 years, female) of very early idiopathic portal hypertension. During an examination for in situ uterine cervical cancer, splenomegaly and hypersplenism were incidentally found. CT and MRI showed a nonatrophic liver with dilated portal veins and marked splenomegaly. The portal venous blood flow was increased, while portal venous blood pressure was not high. The spleen (1,220 g) showed hyperplasia of white pulp and congestion. The lobular architecture of the liver was well-preserved, and the subcapsular regions were not atrophic or dropped out. The portal tracts were not fibrotic, and portal veins were neither stenotic nor sclerotic. Instead, lymphoid cell infiltrations were found in about half the portal tracts, and there was subendothelial mononuclear cell infiltration of small portal vein branches. The hepatic lobules showed non-specific reactive change. This case suggests that early hepatic changes recognizable histologically in this disease are lymphoid cell infiltration of the portal tract and of subendothelial regions of portal vein branches, and nonspecific lobular hepatitis. These hepatic changes, as well as marked splenomegaly, may represent an altered immunophenomenon of this disease.

Median nerve compression by a radially inserted palmaris longus tendon after release of the antebrachial fascia: A complication of carpal tunnel release.

We describe a case that had recurrent median nerve compression after release of the antebrachial fascia in carpal tunnel release. The nerve was compressed by a palmaris longus tendon that was inserted radially into the thenar fascia. After decompression (detachment of the tendon) the patient had symptom relief. Release of the antebrachial fascia in the presence of this tendon variant carries a risk of median nerve compression by the tendon.

Ultrasonography in the diagnosis of ulnar tunnel syndrome caused by an occult ganglion.

We report a case of ulnar tunnel syndrome caused by an occult ganglion which was diagnosed preoperatively by ultrasonography.

[Preoperative bronchial arterial infusion of docetaxel for squamous cell carcinoma of the lung--clinical evaluation and correlation between anti-tumor effect and apoptosis].

As an induction therapy for advanced lung cancer, a patient received bronchial arterial infusion of 30 mg/m2 of docetaxel (TXT). Two weeks after infusion, although the tumor size was not reduced, the central necrotic area was significantly increased. There were no adverse effects. Nineteen days after infusion, we performed a right lower lobectomy and lymph nodal dissection. The resected specimen was examined by immunohistochemistry and biochemistry. The anti-tumor effects were evaluated clinically and the correlation between arterial TXT infusion therapy and apoptosis was studied. Hematoxylin and eosin staining demonstrated moderately differentiated squamous cell carcinoma associated with central necrosis, and cells with chromatin condensation scattered in both the necrotic areas and the margin of this area. It is thought that these findings were the result of this therapy, rather than just the self-necrosis seen in an untreated group.

The synergistic effects of hyperthermia and anticancer drugs on induction of apoptosis.

We studied the synergistic effects of hyperthermia and anticancer drugs on induction of apoptosis in lung cancer cells (LK-2 and LU-65A) using in situ end-labeling of DNA, the DNA fragmentation assay, and transmission electron microscopy. A few apoptotic cells were detected only when both cell lines were heated at relatively high temperature (44 degrees C). Moderate numbers of apoptotic cells were observed when both cell lines were incubated with high concentrations (30 or 40 microM) of anticancer drug. Compared with hyperthermia or anticancer drug alone, the combined treatment induced many apoptotic cells in both cell lines, even in the cells treated with lower concentrations (6 or 8 microM) of anticancer drugs following mild hyperthermia (43 degrees C). In regard to kinetics of apoptotic cells induced by treatment, the maximum induction of apoptosis by the combined treatment was higher than that of hyperthermia or anticancer drug alone in both cell lines, although the time of the peak of apoptotic index differed among the three treatments. Therefore, "hyperthermo-chemotherapy" may reduce the required dosage of anticancer drug and decrease the temperature of hyperthermia on induction of apoptosis.

Video-assisted blebectomy using a flexible scope and a bleb implement.

We describe herein our new technique of performing video-assisted thoracic surgery (VATS) using a flexible scope and a bleb implement which allows for a reduction in the risk of overlooking small and flat blebs in patients with a spontaneous pneumothorax (SP). This technique was performed in 26 consecutive patients with a SP. A skin incision about 18 mm long was made in the area over which the patients had specified preoperatively, and meticulous observation of the apex and superior segment was performed using a bleb implement. Partial pneumonectomy with endoscopic staplers and laser ablation were carried out for pleural lesions. We found that VATS combined with a flexible scope and a bleb implement resulted in better visibility for detection and removal of any small and flat blebs.

The outer membrane protein I of Pseudomonas aeruginosa PAO1, a possible pollutant of dialysate in hemodialysis, induces cytokines in mouse bone marrow cells.

The finding of outer membrane protein I (OprI) of Pseudomonas aeruginosa in hemodialyzers used by patients with end-stage renal failure led us to study the possible role of OprI as cytokine inducer. However, there are few reports on the biological activity of OprI, because it is difficult to obtain highly purified OprI. In this study, we attempt to establish a procedure for the efficient purification of OprI, which does not include lipopolysaccharide, from the bacterial culture broth, not hemodialyzers, to demonstrate that OprI is a potent cytokine inducer. From bacterial culture broth (1 liter), P. aeruginosa PAO1, which was confirmed previously by the sequence coding, was separated by centrifugation, high-performance liquid chromatography, and disk electrophoresis. Mouse bone marrow cells were stimulated by purified OprI, and the supernatants of the culture were analyzed by several enzyme-linked immunosorbent assay kits. The tumor necrosis factor alpha production stimulated by purified OprI was confirmed and degraded within 24 h. Furthermore, interleukin (IL) 1alpha, IL-1beta, IL-6, and granulocyte-macrophage colony-stimulating factor were also induced by OprI despite the absence of lipopolysaccharide. We conclude that OprI has the potential to induce tumor necrosis factor alpha production in mouse bone marrow cells and that tumor necrosis factor alpha contributes to the induction of inflammatory cytokines, namely IL-1alpha, IL-1beta, IL-6, and granulocyte/macrophage colony-stimulating factor, while lipopolysaccharide has little effect on these cells. These results suggest the presence of a pathway of inflammatory signal transduction triggered by OprI. In addition, OprI is possibly one of the harmful dialysate pollutants in hemodialysis patients besides the well-known lipopolysaccharide.

Insufficiency fracture of the femoral neck after intramedullary nailing.

We report a patient with insufficiency fracture that occurred after intramedullary nailing for a subtrochanteric fracture. Intramedullary nailing is speculated to have increased the stress in the already osteoporotic subcapital region. It therefore should be recognized as a causative factor in insufficiency fracture of the femoral neck. Careful follow-up is needed for patients with this condition.