A simplified protocol for molecular sexing in the emu (Dromaius novaehollandiae).

Male and female emus are nearly identical both as chicks and as adults. Although morphological differences of the internal genital tract can be used to distinguish the sexes, a high degree of diagnostic skill is required for accurate sexing. DNA-based sexing methods are highly accurate and can be used to diagnose sex without requiring a high degree of technical skill. However, conventional PCR-RFLP is time consuming and costly, requiring the digestion of PCR products. In this study, we simplified the protocol for sexing the emu by using multiplex PCR without restriction enzyme treatment. Multiplex PCR based on a W-specific primer, with the commonly designed primer set on both Z and W chromosomes, amplified both 197-bp and 272-bp bands in the female, and only the 272-bp band in the male. Sexing results obtained in this way were completely concordant with results obtained using the conventional PCR-RFLP method. Thus, we succeeded in simplifying the protocol for sexing the emu, and suggest that our protocol improves production efficiency by facilitating rapid pairing and selection of individuals to establish high-quality pedigrees.

[Risk factors of recurrence in resected stage I non-small cell lung cancer].

We reviewed risk factors of recurrence in resected pathological stage I non-small cell lung cancer (I NSCLC). Objective is 229 complete resected I NSCLC in our department. Risk factors of recurrence were carcinoembryonic antigen (CEA), histology, differentiation, lymphatic invasion, blood vessel invasion, pleural invasion and tumor size. By univariate analysis, lymphatic invasion (p=0.009), blood vessel invasion (p=0.008), pleural invasion, p1 (p=0.013), p2 (p=0.001), and tumor size (value of cut off was 2 cm) were significant risk factors of recurrence. By multivariate analysis, blood vessel invasion (p=0.004), pleural invasion (p1 or p2) [p=0.001], were significantly risk factors of recurrence. It was suggested that I NSCLC presenting pathological blood vessel invasion and/or pleural invasion should be recognized as cases with a high risk of recurrence, and a strict follow-up and adjuvant therapy should be in consideration.

[Less invasive surgery for the small size peripheral lung cancer: an indication and a review of VATS (video-assisted thoracic surgery) lobectomy].

According to increment of detection of the "small" size peripheral lung cancer measuring less than 20 mm in the greatest dimension, the "less" invasive VATS lobectomy has became widespread among the general thoracic surgeon. To decide on the indication of VATS lobectomy, the frequency of nodal metastasis and recurrence were examined in relation to size of the tumor. The diagnostic accuracy of nodal status in clinical and pathological examinations was 88% in the group with tumors less than 20 mm, 79% in the 21 to 30 mm group and 63% in the c-T2 group, respectively. Also, recurrencies were experienced more frequently in c-T2 (44%) than in c-T1 (19%). In our department, the VATS lobectomy was introduced under the condition as follows; the clinical stage should be limited to T1N0M0. Intraoperative histological examination of the locoregional lymph nodes is required to avoid the risk of local recurrence. In these limited conditions, the survival rate of VATS lobectomy is slightly superior to that of the standard thoracotomy in a same clinical stage. In the future, the application of this procedure should be investigated in older patients, and in more advanced cases.

[Levofloxacin concentrations in serum, sputum and lung tissue: evaluation of its efficacy according to breakpoint].

The levels of levofloxacin (LVFX) in the serum, sputum and lung tissue were measured by a high-performance liquid chromatography method, and the penetration ratio of LVFX into respiratory tissue was investigated. The subjects of this study were 23 patients under pulmonectomy or brochoscopy. LVFX at the dose of 200 mg was given orally and specimens were collected as follows; serum at 2, 3 and 5 hours after, sputum at 2 hours after, and lung tissue at 3 and 5 hours after the administration, respectively. The mean level of LVFX in lung tissue at 3 hours was 3.91 +/- 2.33 micrograms/g, and those in sputum and in serum at 2 hours were 0.71 +/- 0.63 and 2.08 +/- 1.01 micrograms/ml, respectively. A very strong correlation was demonstrated between the level of LVFX in lung tissue and that in serum (p < 0.0001), but correlation between those in sputum and in serum was not significant. The penetration ratio of LVFX into lung tissue was 217.2% and that into sputum was 4.05%. Based on the results of this study, the breakpoints (BPs) of LVFX for pneumonia and chronic respiratory tract infections were calculated to be 4 micrograms/ml and 1 microgram/ml, respectively. It was concluded that penetration of LVFX into lung tissue was satisfactory, and the tissue level of LVFX exceeded greatly the MIC90s against the typical pathogenic bacteria of respiratory tract infections. Taking the excellent BP for pneumonia, 4 micrograms/ml, into consideration, it was thought that LVFX is an effective antibacterial agent against pneumonia and other respiratory tract infections.

Bradycardia induced by irinotecan: a case report.

Irinotecan chloride (CPT-11) is a new semi-synthetic camptothecin analogue which has encouraging antitumor activity against various malignancies. The major and unique toxicity of CPT-11 is diarrhea. Cardiovascular toxicity is rare and has not been found in clinical trials performed in Japan except for a very few cases of insignificant tachycardiac arrhythmia. We report a case of a 69-year-old man with recurrent colon cancer who suffered from bradycardia induced by infusion of CPT-11. Other toxicities including hematological toxicity and diarrhea were mild. Pharmacokinetic analysis using a limited sampling model revealed that the occurrence of bradycardia did not correlate with the excess of drug exposure. Although all of the cholinergic actions reported in the literature were mild, cardiotoxicity may come to be a clinically significant problem. If the events were examined more thoroughly, the cholinergic effect may be discovered more frequently. To administer CPT-11 safely needs meticulous monitoring not only for hematological toxicity and diarrhea but also for other cholinergic actions including bradycardia.

The influence of relative body weight on toxicity of combination chemotherapy with cisplatin and etoposide.

PURPOSE: This study was conducted to determine whether there was any relationship between the adverse toxicity of combination chemotherapy and clinical values including age, sex, creatinine clearance (Ccr), body surface area and relative body weight. METHODS: Cisplatin at a dose of 80 mg/m2 on day 1 and etoposide at a dose of 100 mg/m2 on days 1, 2 and 3 were given to 42 consecutive patients with solid tumors. All patients had normal major organ function and received uniform hydration therapy. RESULTS: Body Mass Index as a measure of relative body weight was inversely correlated with the percentage decrease in white blood cells (P = 0.0681) and platelet count (P = 0.0115). Body surface area was also inversely correlated with leukopenia (P = 0.0171) and thrombocytopenia (P = 0.0058). In contrast, age, sex and Ccr had no significant relationship with adverse toxicity. CONCLUSIONS: It is concluded that dose adjustment of combination chemotherapy with cisplatin and etoposide according to age or ideal body weight is not appropriate and that a conventional dose modification method based solely on body surface area is probably not sufficient to reduce interpatient variability of cancer chemotherapy. A pharmacokinetic and pharmacodynamic study of combination chemotherapy is warranted to establish the ideal dose modification method.

Clinical and experimental studies of anti-tumoural effects of electrochemical therapy (ECT) alone or in combination with chemotherapy.

The present experimental study shows that anticancer agents may accumulate around electrodes placed in saline and in lung when the electrodes are charged with direct electric current. We also observed that electrochemical therapy (ECT) in combination with chemotherapy was more effective for treatment of localized tumours (human lung cancer PC-13 transplated to nude mice) than was ECT or systemic chemotherapy alone. In a clinical study, 26 patients (27 lesions) received ECT with or without systemic chemotherapy. We observed a decrease in tumour size in more than 70% of the cases, and in two cases the use of ECT alone resulted in complete regression of the tumour. In one case there was an increase in tumour size. The main side-effects observed during treatment of lung cancer was pain during treatment, fever after treatment, and pneumothorax. We conclude that ECT may be effective in controlling localised tumours. The clinical use of ECT is, however, associated with several problems and the mechanism of this treatment has not yet been completely established.

[Endoscopic surgery of airway lesions by Nd-YAG laser treatment].

Between April 1980 and November 1990, we treated 212 cases of airway lesion using an Nd-YAG laser via the fiberoptic bronchoscope. The Nd-YAG laser power output was usually 40 W (20-60 W) delivered in 2 sec. shots. The cases consisted of 98 primary lung cancer, 12 primary tracheal cancer, 53 metastatic airway lesion, 7 benign tumor, and 42 cicatricial and granulomatous lesions. The therapeutic effects of Nd-YAG laser treatment were evaluated based on alleviation of dyspnea, widening of airway, and curative vaporization for therapeutic purposes. Effectiveness was observed in 180 of a total of 212 cases (84.9%). Out of 75 emergency cases in which a lifesaving procedure was performed to widen the airway, effective results were obtained in 70 (93.3%) with dramatic improvement in condition. It was also effective in 90 of 109 cases (82.6%) in which the procedure was performed for staged (palliative) widening of airway. In 55 cases of advanced lung cancer (Stage III or IV, mainly non-small cell cancer) in which palliative widening procedure was performed, one year survival was 44%. In 13 of 18 cases (72.2%) in which the procedure was performed for curative vaporization of invasive cancer, successful results were obtained. In 7 cases of benign tumor in which vaporization was performed as a radical curative procedure, no recurrence was observed in any cases. In 53 cases of metastatic airway lesion, effective results were obtained 48 (90.6%). The primary lesions of these cases consisted of 14 cases of esophageal cancer, 9 cases of lung cancer, 7 cases of colo-rectal cancer, 7 cases of thyroid cancer, and 16 others.(ABSTRACT TRUNCATED AT 250 WORDS)

[Clinical studies on reconstruction procedure of total layer chest wall resection].

The results of clinical studies on 16 reconstruction procedure after total layer chest wall resection in 14 cases of malignant tumor of the chest wall were reported. The 14 cases consisted of two cases with recurrent primary chest wall tumor, two cases of primary breast cancer, seven cases of recurrent breast cancer, and others. The reconstruction procedure after total layer chest wall resection was conducted using only various myocutaneous flaps (eight cases using latissimus dorsi of the resected side, three cases using the abdominitis of the resected side, three cases using latissimus dorsi of the non-resected side, and two cases using a pectoralis major myocutaneous flap of the non-resected side). reconstruction only using a myocutaneous flap proved to be satisfactory for preventing early stage postoperative respiratory distress and maintaining the stability of the chest wall and respiratory function during prolonged observation. Namely, use of myocutaneous flap is the best approach of reconstruction the chest wall after total layer chest wall resection. We confirmed that reconstruction with latissimus dorsi myocutaneous free flap of the non-resected side with microvascular anastomosis of thoracodorsal vessels was useful for posterior chest wall tumors invading the latissimus dorsi muscle. Also, our results demonstrated the insertion of an omental flap under the myocutaneous flap was useful for cases with secondary chest wall infection or vascular damage caused by preoperative high dose irradiation.

[A case report of mediastinal malignant lymphoma with long survival following combination chemotherapy using adriamycin, vincristine and cyclophosphamide].

Mediastinal B-cell malignant lymphoma of a 22-year-old female was successfully treated by combination chemotherapy including Adriamycin, Vincristine and Cyclophosphamide. She suffered from dyspnea and axillary tumor in September 1984. Roentgenological examination revealed a large anterior mediastinal tumor. Biopsy of the axillary tumor yielded a diagnosis of metastatic undifferentiated carcinoma from thymus by hematoxylin and eosin. Radiotherapy and chemotherapy including CDDP and ACNU resulted in a symptom-free period of only 2 months. Superior vena cava syndrome and massive pleural effusion recurred. Salvage chemotherapy including Adriamycin, Vincristine and Cyclophosphamide resulted in rapid therapeutic effect. Six courses of chemotherapy were administered, and she is alive and well 4 years after the first salvage chemotherapy. A definitive diagnosis of B-cell lymphoma was made after review of biopsy specimens using immunohistochemical procedures. To select adequate treatment for mediastinal malignant lymphoma, reliable diagnostic procedures including immunohistochemistry are needed. Intensive chemotherapy with appropriate drugs may obtain good response even in advanced cases, such as this.

[Direct current therapy for local control of malignant tumors].

Sixteen malignant tumors of soft tissue (15 cases) were treated with low voltage direct current (DC therapy) with or without systemic chemotherapy. Two platinum or stainless steel electrodes were placed in the tumor and around 10 volts of direct current was passed for 1 hour. In cases in which chemotherapy was performed, single or combined anticancer agents consisted of ADM and BLM were administered simultaneously with DC therapy by systemic route. Ten cases (11 tumors) out of 15 cases received no other local treatment. Histological examination was performed in 6 of 11 lesions, and in all lesions pathological therapeutic effects were recognized. Decrease in tumor size was observed in 9 lesions. In 2 cases the tumors disappeared completely after DC therapy only. In one case receiving DC therapy with chemotherapy, the tumor decreased dramatically and became resectable. Main complications were slight pain during treatment and slight fever after treatment, but special treatment for these complications was not necessary. Our results suggested that DC therapy with or without systemic chemotherapy was useful for local control of malignant tumors. And also if DC therapy had been performed simultaneously with chemotherapy the therapeutic effects seemed to be more emphasized than single use of these therapies.

Immunohistochemical and immunoblot analyses of ras-p21 expression in lung carcinomas.

Immunohistochemical and immunoblot analyses were performed on 49 cases of surgically resected primary lung carcinoma to examine the expression of ras oncogene product using monoclonal antibodies to ras-p21. Two different monoclonal antibodies, NCC-RAS-001 and RAP-5 were used for immunohistochemical study. Cancer cells of 16 cases (33%) and 15 cases (31%) were strongly positive for NCC-RAS-001 and RAP-5, respectively. The staining pattern of antibodies was heterogenous among cancer cells, even in the same case. Among various histologic type of lung cancers, squamous cell carcinomas and well-differentiated adenocarcinomas had a tendency to react more intensively than other histologic types of carcinoma. Immunoblot analysis using monoclonal antibody NCC-RAS-004 revealed the presence of ras-p21 not only in cancer tissues but also in non-cancerous tissues in all cases analysed. In 13 cases (27%), cancer tissue expressed more than twice as much ras-p21 as non-cancerous tissues. Our study showed that the over-expression of ras-p21 in lung carcinomas compared with non-cancerous tissues was a relatively common phenomenon, especially in squamous cell carcinomas and adenocarcinomas.