Evaluation of genotoxicity and subchronic toxicity of standardized rose hip extract.

Rose hip is the fruit of the rose plant, which is widely used in food, cosmetics and as a traditional medicine. Therefore, rose hip is considered safe and has a sufficient history of consumption as food. However, few studies have reported on the safety of rose hip extracts in toxicological analyses. Thus, to evaluate the safety of rosehip polyphenol MJ (RHPMJ), an aqueous ethanol extract standardized with the trans-tiliroside content, we performed genotoxicity and 90-day repeated oral dose toxicity studies in compliance with the Organisation for Economic Co-operation and Development-Good Laboratory Practice. RHPMJ did not induce gene mutations in reverse mutation tests of Salmonella typhimurium TA98, TA100, TA1535, TA1537 and Escherichia coli WP2 uvrA strains and did not induce chromosomal aberrations in cultured Chinese hamster lung (CHL/IU) cells. Moreover, micronucleus tests using rat bone marrow showed RHPMJ had no micronucleus-inducing potential. Finally, 90-day repeated oral dose toxicity studies (100-1000 mg/kg) in male and female rats showed no treatment-related toxicity in rats. These data indicate that the RHPMJ had no genotoxicity and a no-observed-adverse-effect level greater than 1000 mg/kg in rats.

Validity of putative calcium binding loops of photoprotein aequorin.

Three peptides containing the putative Ca2+ binding loops, I, II and III, respectively, of a photoprotein, aequorin, from jellyfish Aequorea victoria were synthesized by a solid-phase procedure. The peptides bound Ca2+ with dissociation constants of 10(-3) to 10(-4) M, providing evidence for the assumption that Ca2+ binding loops are actually responsible for the binding of Ca2+. When the highly conserved 6th glycine residue in the 12-residue loops was replaced by arginine, no large effect was observed on Ca2+ binding. Exposure to a hydrophobic environment and the binding of Ca2+ brought about conformational changes to the peptides.

Concurrent daily carboplatin and accelerated hyperfractionated thoracic radiotherapy in locally advanced nonsmall cell lung cancer.

PURPOSE: This study was designed to evaluate the feasibility and efficacy of accelerated hyperfractionated thoracic radiotherapy concurrently combined with daily carboplatin in patients with locally advanced, unresectable nonsmall cell lung cancer. METHODS AND MATERIALS: Thirty-one patients with locally advanced nonsmall cell lung cancer were treated with continuous course, twice daily thoracic radiotherapy (1.5 Gy each) to a total of 60 Gy over 4 weeks. Carboplatin (25 mg/m2) i.v. was given immediately before each morning thoracic radiotherapy. Blood samples were taken to measure the blood free platinum pharmacokinetics on day 1. RESULTS: All 31 patients completed the protocol treatment without delay. The median age was 73 years, and the majority had Stage IIIA (32%) or IIIB (48%) disease. Major acute toxicity (> or = Grade 3) included 17 patients (55%) with leukopenia, 5 patients (16%) with thrombocytopenia, and 7 patients (23%) with esophagitis. One possible treatment-related death due to diffuse pneumonitis was observed. There were three complete responses (CRs) and 23 partial responses (PRs) in the radiation field, for a response rate of 84%. The relapse pattern was predominantly loco-regional, and the median survival time was 9.8 months. The area under the plasma level-time curve (AUC) of free platinum correlated significantly (r = -0.41, p = 0.04) with the surviving fraction of leukocytes, but not with the severity of the esophagitis. Responders had significantly (p = 0.04 by Welch's t-test) higher AUCs than nonresponders. CONCLUSIONS: This combination therapy was feasible and efficacious against locally advanced nonsmall cell lung cancer. Although long-term local control still remains unsatisfactory, pharmacokinetic data are suggestive of a role for platinum in enhancing the radiation effect.

A randomized controlled trial of sulfamethoxazole/trimethoprim plus norfloxacin versus sulfamethoxazole/trimethoprim alone for the prophylaxis of bacteria infection during chemotherapy for lung cancer.

The efficacy of the prophylactic administration of sulfamethoxazole/trimethoprim (ST) plus norfloxacin (NFLX) versus ST alone to prevent the development of bacterial infection during chemotherapy-induced leukopenia was compared in patients with lung cancer. Patients who underwent systemic chemotherapy were randomized into one of the prophylactic regimens when grade 3 or 4 leukopenia occurred. Prophylactic treatment was performed on 133 courses of leukopenia in 75 patients and the efficacy was evaluated on 127 of those courses after excluding those patients who demonstrated a fever within 24 h from the start of the prophylaxis. The number of patients who had leukopenia associated fever was two out of 63 (3.2%) with the ST plus NFLX regimen and 10 out of 64 (15.6%) with ST alone; the difference was statistically significant. The prophylactic use of ST plus NFLX was thus found to be more useful than ST alone for the treatment of chemotherapy-induced leukopenia in patients with lung cancer.

Prediction of carboplatin clearance calculated by patient characteristics or 24-hour creatinine clearance: a comparison of the performance of three formulae.

PURPOSE: Carboplatin doses can be individualized using the formula of Calvert et al. (Calvert formula) dose (mg) = area under the plasma concentration versus time curve (AUC) x [glomerular filtration rate (GFR) + 25]. Creatinine clearance (Ccr), either measured by the 24-h method or calculated by the formula of Cockcroft and Gault [Cockcroft-Gault (CG) formula], is often substituted for the GFR. The CG formula is based on patient weight, age and sex, and the serum creatinine (Cr) concentration. Another method for predicting carboplatin clearance (CL) using patient characteristics has also been proposed by Chatelut et al. (Chatelut formula). This study was undertaken to evaluate the performance of the three formulae in predicting standard- and low-dose carboplatin pharmacokinetics. METHODS: A total of 52 patients with advanced lung cancer were enrolled in this pharmacokinetic study; 37 received standard-dose carboplatin and 25 received low-dose carboplatin. The Cr concentration was measured using an enzymatic assay. The three formulae were used to predict carboplatin CL. The median absolute percent error (MAPE) for each formula was evaluated by comparing the calculated and observed CL. For comparison of AUCs, free platinum plasma concentrations were measured at intervals up to 24 h after carboplatin administration. AUCs were determined and compared with predicted values. RESULTS: In the standard-dose carboplatin group, the MAPEs for the prediction of carboplatin CL from the 24-h Calvert, CG-Calvert and Chatelut formulae were 13%, 12% and 23%, respectively. In the low-dose carboplatin group, the corresponding MAPEs were 27%, 18% and 44%, respectively. Observed standard-dose carboplatin AUCs after aiming for target AUCs of 5 and 6 mg x min/ml using the Calvert formula based upon the 24-h Ccr were 5.3+/-0.8 and 5.9+/-0.8, respectively, indicating a small and acceptable bias compared with that predicted from the dosing formula. CONCLUSIONS: The pharmacokinetics of standard-dose carboplatin were accurately predicted by the Calvert formula based upon either 24-h or CG-calculated Ccr, but not by the Chatelut formula. Either CG-calculated or 24-h Ccr can be substituted for the GFR in the Calvert formula for the determination of individual doses. The poor predictability of the Chatelut formula found in this study might be the result of a differences in either the Cr assay or the patient population. Therefore, formulae which attempt to estimate GFR are not necessarily valid if either the Cr assay or the patient population is changed.

Legionnaires' disease diagnosed by bronchoalveolar lavage.

A 51-year-old woman who had been on steroid therapy for systemic lupus erythematosus (SLE) developed a high fever 3 days after visiting a hot spring resort. Chest X-ray films revealed an interstitial, pneumonia-like shadow in the left lung field, which increased rapidly with a worsening of her symptoms. She died of multiple organ failure one week after the onset of the pneumonia. Although the serum antibody titer was negative, Legionella pneumophila was recovered from her bronchoalveolar lavage (BAL) fluid. BAL seems to be a useful method to diagnose Legionnaires' disease.

Direct identification of lung cancer cells producing granulocyte colony stimulating factor with monoclonal anti-G-CSF antibodies.

This is a case report of an 82-year-old man with squamous cell lung cancer which produced G-CSF. The leukocyte cell count was elevated to 94,400/mm3. The serum G-CSF concentration was also elevated to 351 pg/ml. This is the first report to demonstrate, by application of immunohistochemical staining techniques directly to tumor tissue obtained at autopsy, squamous carcinoma cells which produce G-CSF. The tumor cells were stained with monoclonal anti-G-CSF antibodies.

Response to combined modality treatment in a five-year survivor of extensive small cell lung cancer with severe complications.

We present a rare case of a five-year survivor of small cell lung cancer with severe complications who responded to combined modality treatment. Prior to initial chemotherapy, he experienced severe complications including sepsis, pneumonia, ileus, and a performance status of 4. He was treated with an ileus tube and IVH, and was managed by mechanical ventilation for four days. After his general condition improved, he received combination chemotherapy of carboplatin, with the target area under the plasma concentration versus the time curve (AUC) of 5 mg x min/ml day 1, and etoposide (80 mg/m2) on days 1, 2, 3 for four courses, and complete remission (CR) was obtained. Six months later, systemic relapse occurred, but he achieved complete remission again with nine courses of CODE (cisplatin, vincristine, adriamycin, and etoposide) chemotherapy and sequential chest radiotherapy. Five years after the initial chemotherapy, the patient is alive and disease free.

Multifocal electroretinograms in normal subjects.

Multifocal electroretinography (ERG), developed by Sutter in 1992, is a method of recording the spatial distribution of focal ERG in a short time period using multi-input stimulation. Using this technique, we can detect the spatial extent and severity of damage to the macula. In this study, we recorded multifocal ERGs from 20 eyes of 20 normal subjects and analyzed the topographical properties of responses. In every subject, a negative wave followed by a positive wave could be recorded and we named them the N1-wave and the P1-wave, respectively. The amplitudes of the N1-wave and the P1-wave were the largest in the fovea, and they became smaller with eccentricity. In the P1-wave amplitude, the greatest intersubject variability was observed at the fovea. The N1 and P1 latencies were shorter in the upper retina than in the lower retina. The amplitude was larger in the upper retina than in the lower retina, which suggests the functional superiority of the upper retina. There was no statistical difference in latency and amplitude between the nasal and the temporal retina. We found no statistical difference between the responses of the papillomacular bundle and those of the temporal retinal area. The mapping obtained by multifocal ERG was useful as objective perimetry.

[Multi-focal electroretinograms in normal subjects].

The multi-focal electroretinogram (multi-focal ERG), developed by Sutter (1992), is a method of recording the spatial distribution of focal ERG in a short time using multi-input stimulation. Using this technique, we can detect the spatial extent and severity of damage to the macula. In this study, we recorded multi-focal ERGs from 20 eyes of 20 normal subjects and analyzed the topographical property of responses. In every subject, a negative wave followed by a positive wave could be recorded and we named them the N1-wave and the P1-wave, respectively. The amplitudes of the N1-wave and the P1-wave were the largest in the fovea and they became smaller with eccentricity. In P1-wave amplitude, the greatest inter-subject variability was observed at the fovea. The N1 and P1 latencies were shorter in the upper retina than in the lower retina. The amplitude was larger in the upper retina than in the lower retina, which suggests functional superiority of the upper retina. There was no statistical difference of latency and amplitude between nasal and temporal retina. We found no statistical difference between the responses of the papillomacular bundle and those of the temporal retinal area. The mapping obtained by multi-focal ERG was useful as objective perimetry.

[A case of pulmonary cavernous hemangioma: immunohistological examination revealed its endothelial cell origin].

A 29-year-old woman was referred to our hospital for an abnormal shadow on chest roentgenogram. She had no symptom but its shadow was pointed out about 20 years ago. Chest CT scan showed a sharply demarcated homogenous mass measured 10 x 10 mm in the S4 segment. The tumor was resected with video-assisted thoracoscopic surgery under CT-guided marking wire inserted. Microscopic examination suspected cavernous hemangioma. Immunohistological staining demonstrated most lining cells of the cavernous structure stained positively for von Willebrand factor antibody, which suggests that this tumor was associated with endothelium. Then we decided that this tumor was pulmonary cavernous hemangioma.

[Regrowth of bullae around the staple-line is one of the causes of postoperative recurrence in thoracoscopic surgery for spontaneous pneumothorax].

Since August 1994, we have performed 65 thoracoscopic surgeries for spontaneous pneumothorax. Our operative method is partial resection of the lung with bullae using endoscopic autosuture. Three patients underwent reoperation because of persistent air leakage in one case and postoperative recurrences in two cases. In addition, we performed a second operation in one case that had undergone thoracoscopic surgery for spontaneous pneumothorax in another hospital. At the second operation, regrowth of bullae was found at the edge of staple-line in three of the 4 cases, and in one of these cases, such regrown bullae seemed to be a cause of recurrence of pneumothorax. The regrowth of the bullae was caused by incomplete resection at the initial surgery in one case. Such incomplete resection and oversight were not noticed in the review of the videotape of the initial surgeries in two cases. Reinforcement of the staple-line is necessary for prevention of recurrence of pneumothorax.

[Pulmonary histoplasmosis exhibiting solitary pulmonary nodule resected by thoracoscopic surgery: a case report and review of the Japanese literature].

We reported a case of pulmonary histoplasmosis showing solitary nodular shadow. A 43-year-old man was referred to our hospital because of an abnormal shadow on chest X-ray films during a routine checkup. He had traveled to Honduras for 7 days. Chest computed tomographic (CT) scans showed a 13 x 12 mm nodular shadow with unclear margin in the left upper lobe (S3). Both transbronchial lung biopsy and CT guided transcutaneous needle biopsy failed to yield a definitive diagnosis. Thoracoscopic resection of the nodule was performed due to suspicion of lung cancer. Pathologically, the nodule displayed central caseous necrosis with many round yeast-like fungi. The fungi measured 3 to 4 microns in diameter and were well-stained by Grocott stain. Immunohistochemical staining was positive for anti-Histoplasma capsulatum antibody, resulting in the final diagnosis of pulmonary histoplasmosis. The patient's postoperative course was uneventful, and no recurrence was observed. Histoplasmosis is a rare disease in Japan. However, it is important to keep imported infectious diseases in mind when examining and treating patients who have a history of travel abroad.

Occurrence of an allosteric transition in the modification of papain with L-1-acetyl-2,3-dihydropyrrolo[2,3-b]-indole-2-carboxamide.

When papain was reacted with L-1-acetyl-2,3-dihydropyrrolo[2,3-b]indole- 2-carboxamide at pH 8.0, inactivation occurred accompanied by modification of Cys-25 in the active site. Plots of pseudo first-order rate constants against the reagent concentrations yielded an anomalous sigmoidal curve, suggesting that papain responded to this reagent in an allosteric manner. This is supported by the fact that the presence of a moderate concentration (a twenty-fold molar excess) of N alpha-acetyl-L-tryptophanamide over papain accelerated the inactivation.

Predicting the need for hospital admission in patients with acute bronchial asthma.

Historical data, physical findings, pulmonary function, arterial blood gases, and subjective degree of dyspnea rated on a modified Borg scale were correlated with eventual requirement of hospitalization in 83 episodes of acute asthma attacks of 70 adult patients. Among the pretreatment data, only pulse rate remained significant by a multivariate analysis to predict hospitalization. For patients who had apparently been successfully treated in the emergency room and discharged home, residual degree of subjective dyspnea was the only significant variable chosen by a linear discriminant function to predict the eventual need for hospitalization, with a sensitivity of 75% and a specificity of 78%. We concluded that careful clinical evaluation still remains the best available diagnostic tool in the care of acute asthma.

A double-blind, placebo-controlled trial of the thromboxane synthetase blocker OKY-046 on bronchial hypersensitivity in bronchial asthma patients.

We investigated the effect of thromboxane (TX) synthetase inhibitor, OKY-046, on bronchial hypersensitivity in 16 asthmatics by a double-blind, placebo-controlled, crossover trial. Bronchial sensitivity to methacholine was measured by Astograph. Blood samples were taken to measure plasma levels of TX metabolites. No significant differences of forced expiratory volume in 1 sec (FEV1), bronchial sensitivity, or bronchial reactivity were observed after OKY-046 administration, compared to baseline or after placebo. However, responders showed a significant decrease in the plasma TXB2/6-keto-PGF1alpha ratio as compared to nonresponders. Our data failed to confirm an inhibitory effect of OKY-046 on bronchial hypersensitivity, but suggested the importance of its therapeutic dose monitoring.

A phase I clinical and pharmacologic study of a carboplatin and irinotecan regimen combined with recombinant human granulocyte-colony stimulating factor in the treatment of patients with advanced nonsmall cell lung carcinoma.

BACKGROUND: This Phase I study was designed to determine the toxicity and efficacy of a carboplatin and irinotecan (CPT-11) regimen with recombinant human granulocyte colony-stimulating factor (rhG-CSF) support for patients with advanced nonsmall cell lung carcinoma. METHODS: Treatment consisted of carboplatin administered intravenously (i.v.) on Day 1 plus CPT-11 i.v. on Days 1, 8, and 15. The carboplatin dose was calculated using Calvert's formula, where the target area under the plasma concentration versus the time curve (AUC) was 5 or 6 mg x min/mL. rhG-CSF (2 microg/kg) was administered daily, except on Days 1, 8, and 15, until the leukocyte count exceeded 20,000/mm3 (10,000/mm3 after Day 16). Cycles were repeated every 4 weeks. Groups entered the trial at escalating CPT-11 and carboplatin dose levels of 60 mg/m2 and 5 mg x min/mL, 70/5 and 60/6. RESULTS: Twenty-one patients were enrolled in this study, of whom 20 were assessable for toxicity and therapeutic efficacy. Two of 6 patients experienced Grade 4 diarrhea at the 70/5 dose level, suggesting that this was the maximum tolerated dose (MTD). However, the 60/6 dose level was included because toxicities were very mild at the 60/5 dose level. At the 60/6 dose level, 1 of 6 patients experienced severe, life-threatening toxicity. Therefore, subsequent dose escalation was stopped and the study terminated. There were 7 responses (35%) among the 20 patients. At the 60/6 dose level (n=5), the observed carboplatin AUC after aiming for a target AUC of 6 was 5.9+/-0.9 mg x min/mL, as expected, although the AUCs of both CPT-11 and its active metabolite, SN-38, were lower than expected. CONCLUSIONS: The recommended doses for Phase II studies are 60 mg/m2 of CPT-11 and a target AUC of 5 mg x min/mL for carboplatin, plus rhG-CSF. The combination of AUC-based carboplatin and CPT-11 with rhG-CSF support appears to be an active regimen in the treatment of patients with NSCLC.