Changes in peripheral anti-Müllerian hormone concentration and their relationship with testicular structure in beef bull calves.

The aim of this study was to clarify the time-course of changes in anti-Müllerian hormone (AMH) and testosterone (T) concentrations in peripheral blood and to determine the relationships between blood AMH concentration and testicular development during the early postnatal and prepubertal periods in beef bull calves. A total of 17 Japanese Black bull calves were enrolled in this study. The wk in which the calf was born (within 6 d after birth) was defined as M 0. Blood samples were taken once in every mo from M 0 to M 6 from each bull calf, and plasma AMH and T concentrations were determined. Of the 17 calves, 10 were castrated at 6 mo of age (prepuberty) and the right testis was histologically examined. Plasma AMH concentration (means ± SE) at M 0, 1, and 2 were 123.5 ± 9.8, 189.6 ± 18.7, and 254.6 ± 14.1 ng/mL, respectively. From M 0 through M 2, plasma AMH concentration was significantly greater each mo than in the previous mo (P < 0.05); however, plasma AMH concentration significantly decreased over the last 3 mo of the study (P < 0.05). The average age at which plasma AMH concentration was the highest was 2.3 ± 0.1 mo of age. Plasma T concentration significantly increased from M 0 (0.18 ± 0.02 ng/mL) until M 6 (6.52 ± 1.41 ng/mL). Plasma AMH and T concentrations at M 4, 5, and 6 were significantly negatively correlated (P < 0.05). Linear regression did not reveal a significant relationship between Sertoli or Leydig cell numbers and plasma AMH or T concentrations, respectively. In conclusion, blood AMH concentration peaks at 2 mo of age and is negatively correlated with blood T concentration from 4 to 6 mo of age. Although prepubertal blood AMH or T concentrations did not reflect Sertoli or Leydig cell numbers at the end of the prepubertal period, blood AMH concentration may be indicative of abnormal Sertoli cells function.

Estrogenic activity of 37 components of commercial sunscreen lotions evaluated by in vitro assays.

Thirty-seven chemical components of commercial sunscreen lotions were evaluated for estrogen agonistic and/or antagonistic activity using two in vitro assays, (1) an ELISA-based estrogen receptor competitive binding assay (ER-ELISA) and (2) a modified yeast two-hybrid estrogen assay, with and without addition of a rat liver preparation, S9 mix. Eleven compounds, most of which were benzophenone derivatives and parabens, showed binding affinity to ER by ER-ELISA without S9 mix. Although the activities of almost all of the compounds were attenuated by addition of S9 mix, 4-octylphenylsalicylate and 2,2'-dihydroxy-4,4'-dimethoxybenzophenone acquired estrogenic activity, suggesting metabolic activation of these compounds. Two benzophenones showed agonistic activity in the yeast two-hybrid assay without S9 mix. The activity of one of these was reduced by S9 treatment and a further two benzophenones was activated. Eight parabens were active in this assay without S9 exposure, but their activities were eliminated by S9 treatment. Benzophenones with para-phenolic hydroxyl groups and parabens with branched and/or longer linear chains were generally more potent in both bioassays. In addition, weak antagonistic activity of 4-t-butylphenyl-salicylate, 2-ethylhexyl 4-dimethylaminobenzoate and (+/-)-alpha-tocopherolacetate was observed with S9 treatment. In vivo testing of the compounds reported here to have estrogen agonistic and antagonistic activities is required to confirm their effects at an organismal level.

Inhibition of corneal inflammation by an acylated superoxide dismutase derivative.

Superoxide radicals and their metabolite(s) have been postulated to play an important role in the pathogenesis of inflammation. Hence, superoxide dismutase (SOD) has been used to reduce tissue injury caused by reactive oxygens. However, protection of the cornea and other ocular tissues from oxygen toxicity could not be achieved by administering SOD presumably due to its unfavorable in vivo behavior. To scavenge superoxide radicals on the outer surface of corneal epithelial cells, the authors synthesized an acylated SOD derivative (AC-SOD) by linking capric acid. When instilled into rabbit eyes, a significant amount of AC-SOD remained bound to the corneal surface for a fairly long time. Intracorneal injection of lipopolysaccharide (LPS) triggered infiltration of polymorphonuclear leukocytes (PMNs) to the cornea and induced severe keratitis. Topical administration of AC-SOD to the LPS-treated cornea markedly inhibited the infiltration of PMNs and suppressed the occurrence of keratitis. Under identical conditions, topically administered SOD was rapidly removed by tears and, hence, did not inhibit LPS-induced keratitis. When the number of PMNs in the systemic circulation was reduced by intravenous administration of hydroxyurea, LPS-induced keratitis was inhibited markedly. These results indicate that superoxide radicals and circulating PMNs might play a critical role in LPS-induced keratitis.

Pathogenesis of serratial infection: activation of the Hageman factor-prekallikrein cascade by serratial protease.

A serratial protease with an apparent molecular weight of 56,000 (56K protease), which had been purified from the culture supernatant of a strain of Serratia marcescens isolated from a corneal lesion of a human eye [Matsumoto, K. et al. (1984) J. Bacteriol. 157, 225-232], greatly enhanced vascular permeability when injected into guinea pig skin. The 56K protease, which requires zinc ion for activity, was found to possess plasma kallikrein-like properties in vitro as judged by (i) preferential amidolysis of carbobenzoxy-Phe-Arg-4-methylcoumaryl-7-amide and Pro-Phe-Arg-4-methylcoumaryl-7-amide, which are known substrates for plasma kallikrein; (ii) release of kinin from high-molecular-weight kininogen; and (iii) prompt activation of Hageman factor followed by generation of kallikrein from plasma prekallikrein. These results suggest that the 56K protease enhances vascular permeability through activation of a Hageman factor-kallikrein-kinin pathway in vivo, and this molecular process appears to be a rational mechanism of enhancement of permeability and serratial pathogenesis.

Correlation of binding sites for diisopropyl phosphorofluoridate with cholinesterase and neuropathy target esterase in membrane and cytosol preparations from hen.

To find new putative target(s) for organophosphorus induced delayed neurotoxicity (OPIDN), we investigated the biochemical and pharmacological characteristics of [3H] diisopropyl phosphorofluoridate (DFP) binding to membrane and cytosol preparations from the brain and spinal cord of hens. Specific [3H]DFP binding was determined by subtracting non-specific binding from total binding. The binding sites of [3H]DFP, an organophosphate that induces OPIDN, were found not only on membrane but also in cytosol. Reduction of subsequent ex vivo specific [3H]DFP binding by in vivo pretreatment with unlabeled DFP was found in cytosol, not membrane. The reduced binding lasted to the onset of OPIDN, especially in spinal cord. These results suggest that the specific DFP binding sites in cytosol, rather than on membrane, are the most important with regard to the initiation of OPIDN. Inhibitors of cholinesterase (ChE) and neuropathy target esterase (NTE) other than DFP did not affect specific [3H]DFP binding to either membranes or cytosol in vivo. Additionally, inhibition of the activities of these esterases by these compounds was not consistent with either the degree of inhibition of the [3H]DFP binding or a time-dependent manner of OPIDN. These results suggest that DFP binding site(s) involved in the initiation of OPIDN may be different from the active sites of ChE and NTE.

A comparative study of binding sites for diisopropyl phosphorofluoridate in membrane and cytosol preparations from spinal cord and brain of hens.

Biochemical events in the initiation of organophosphorus induced delayed neurotoxicity (OPIDN) are not well understood. To find new putative target(s) for OPIDN, we investigated the biochemical and pharmacological characteristics of [3H] diisopropyl phosphorofluoridate (DFP) binding to membrane and cytosol preparations from the brain and spinal cord of hens in vitro. [3H]DFP binding to both preparations was determined by the specific binding obtained by subtracting non-specific binding from total binding. The specific binding sites of [3H]DFP were found not only on membrane but also in cytosol. Kd values were higher and Bmax values were lower in cytosol than in membrane. Moreover, the Kd values in both membrane and cytosol preparations from spinal cord were lower than those of brain. The Bmax values in membrane and cytosol were similar between brain and spinal cord. The specific binding to both preparations was markedly displaced by unlabeled DFP. The specific binding of DFP to the membrane was highly or partly displaced by organophosphorus compounds (OPs) or a carbamate, respectively. However, both the OPs and the carbamate had considerably weaker blocking effects on the specific binding of DFP to cytosol. None of the compounds known to interact with neuropathy target esterase (NTE) had a strong blocking effect on the specific binding of DFP to either membrane or cytosol. These results show that the specific binding of DFP to the membrane may be binding with cholinesterase (ChE). However, cytosol, especially in spinal cord, may have DFP binding sites other than ChE and NTE.

Enhancement of vascular permeability upon serratial infection: activation of Hageman factor--kallikrein--kinin cascade.

A zinc dependent serratial 56K protease caused enhancement of vascular permeability followed by edema formation when injected into the guinea pig peripheral cornea, the subconjunctival space, or the skin. Because this enhancement was not affected by antihistamine, involvement of the kinin-generating system in this permeability enhancement was investigated. The 56K protease induced permeability much greater extent than that by bradykinin on weight basis, and more so on molar basis. The phenomenon was inhibited by soybean trypsin inhibitor, a well known inhibitor of plasma kallikrein, and also by corn trypsin inhibitor, which is the best inhibitor of the activated Hageman factor. In vitro experiments using numbers of synthetic peptide substrates, the 56K protease exhibited a similar substrate specificity to that of plasma kallikrein. Kallikrein is a known endogenous activator of Hageman factor. The enhancement by 56K protease was greatly augmented by inhibition of kininase II with Glu-Trp-Arg-Pro-Gln-Ile-Pro-Pro-OH (SQ 20,881), suggesting generation of bradykinin. Thus, these results indicate that the enhancement of vascular permeability induced by the 56K protease is caused by an activation of Hageman factor by 56K protease followed by subsequent activation of cascade amplification, and resulted in kinin generation in vivo.

Recent progress in the neurotoxicology of natural drugs associated with dependence or addiction, their endogenous agonists and receptors.

Nicotine in tobacco, tetrahydrocannabinol (delta 9-THC) in marijuana and morphine in opium are well known as drugs associated with dependence or addiction. Endogenous active substances that mimic the effects of the natural drugs and their respective receptors have been found in the mammalian central nervous system (CNS). Such active substances and receptors include acetylcholine (ACh) and the nicotinic ACh receptor (nAChR) for nicotine, anandamide and CB1 for delta 9-THC, and endomorphins (1 and 2) and the mu (OP3) opioid receptor for morphine, respectively. Considerable progress has been made in studies on neurotoxicity, in terms of the habituation, dependence and withdrawal phenomena associated with these drugs and with respect to correlations with endogenous active substances and their receptors. In this article we shall review recent findings related to the neurotoxicity of tobacco, marijuana and opium, and their toxic ingredients, nicotine, delta 9-THC and morphine in relation to their respective endogenous agents and receptors in the CNS.

Quantitative examination of taste deficiency due to radiation therapy.

A study was carried out with a Rion TR-03 type electro-gustometer for subjective and quantitative expression of the degree of taste deficiency due to radiation therapy and the effects of treatment. The changes of the electro-gustometric threshold due to irradiation were examined in patients who were divided into three groups: those receiving irradiation of the whole oral cavity, those receiving irradiation of the posterior tongue and the soft palate and those receiving irradiation of the gustatory nervous system. In the group receiving irradiation of the whole oral cavity, the threshold showed a tendency to increase when receiving 10 Gy/5 days, and the threshold became unmeasurable when more than 40 Gy/26 days was given. In the group receiving irradiation of the posterior tongue and the soft palate, almost no change of the threshold in the anterior was observed in spite of increasing irradiation doses. In the group receiving irradiation of the gustatory nervous system, the threshold in the intact hemicephalus did not changed even after irradiation, but a change was found on the pathological side. Based on these results, the authors originated a complication scoring system and a treatment scoring system both of which are divided into five stages. An electro-gustometric examination is used in order to provide effective treatment with a preserving function something which has been strongly desired for radiation therapy of cancer.

Spreading of Serratia marcescens in experimental keratitis and growth suppression by chicken egg white ovomacroglobulin.

We studied the inhibitory effects of chicken egg white ovomacroglobulin, a broad-spectrum protease inhibitor, and a synthetic protease inhibitory peptide, Bz-GFR-O-mercaptoanilide, on the growth of two strains of Serratia marcescens in a synthetic medium. The growth of the virulent protease-producing strain, S. marcescens kums 3958 (kums 3958), was more rapid than that of the strain producing minimal protease, S. marcescens kums NA (kums NA), and kums 3958 was inhibited in a dose-dependent manner by ovomacroglobulin. The synthetic inhibitor also inhibited the growth of kums 3958 weakly. Dose-dependent enhancement of growth of kums NA was observed in the medium treated with the serratial 56 kilo Dalton protease. By immunohistochemical methods using an antibody to the bacteria, the spreading of kums 3958 was also studied in corneal tissue in experimental keratitis of the guinea pig. In vivo, kums 3958 grew locally, and then spread widely by destroying stromal tissue 7-8 hours after intrastromal injection of the organisms (3 x 10(4) colony-forming units). On the other hand, when kums 3958 mixed with ovomacroglobulin was injected into the cornea, the organisms remained locally in the corneal stroma which showed a lower grade of damage. These results indicate that proteases secreted from S. marcescens destroyed corneal stroma, yielding the organisms enough space for further spreading.

Effects of prophylactic irradiation of acute lymphoblastic leukemia in the central nervous system of children.

The development of prophylactic therapy for acute lymphoblastic leukemia (ALL) in the central nervous system in children has resulted in an improvement of the prognosis and prolongation of patients' lives. On the other hand, late irradiation effects have increased with the improvement of the prognosis. Therefore, there is now an important need to lessen these late effects without deteriorating the prognosis. We have investigated the recurrence rate, the survival rate and the cause of death of ALL patients who were divided into a none prophylactic irradiation group and 15 Gy, 20 Gy and 24 Gy irradiation groups. The results show that the effect of prophylactic irradiation has been more effective in the 20 Gy group than that of the 24 Gy group. Furthermore, brain atrophy and leukoencephalopathy, which were investigated by X-ray CT in long-term survivors of post-prophylactic irradiation without recurrence, have been less in the 20 Gy group than in that of the 24 Gy group. This report presents the result that the most favorable dose for prophylactic irradiation for central nervous system leukemia of ALL in children is 20 Gy by a step-up method.

A possible role of nitric oxide formation in the vasodilatation of rabbit ear artery induced by a topically applied Capsaicin analogue.

Effects of topical application of a capsaicin analogue, nonylic acid vanillylamide (NVA, 0.032-10.0 mM) on the arterial diameter in the ear skin were examined in conscious rabbits using a precise dial caliper. In addition, the possibility of nitric oxide (NO) participating in a vasodilatation induced by low concentrations of NVA was tested by an NO synthase inhibitor. At the lowest concentration of NVA (0.032 mM), no significant change in the diameter was observed after external application of the NVA ointment. At concentrations of 0.32 mM or more, NVA produced a significant vasodilator response. However, at higher concentrations of 3.2 and 10.0 mM, -NVA induced substantial shrinkage in the arterial diameter and oedema formation, which was not affected by L-NAME (NG-nitro-L-arginine methyl ester, 3 mg/kg, i.v.), suggesting fluid leakage induced by oedema from the vessels might suppress the vasodilatation. Thus, the concentration-response curve for NVA was bell-shaped. NVA (0.32 mM)-induced vasodilatation was not significantly affected by atropine (1 mg/kg, i.v.) or propranolol (80 micrograms/kg, i.v.). However, the NVA-induced vasodilatation was completely suppressed by an NO synthase inhibitor, L-NAME (3 mg/kg, i.v.) which had no influence on the resting diameter, but not by an inactive stereoisomer, D-NAME (3 mg/kg, i.v.). These findings suggest that vanilloid receptor activation results in the release of sensory neuropeptides, which in turn stimulate the synthesis of endothelial NO which is responsible for the vasodilatation.

Inhibition of collagen-induced platelet aggregation in Japanese black cattle with inherited platelet disorder, Chediak-Higashi syndrome.

Aggregation properties of platelets were examined in Japanese Black cattle with Chediak-Higashi syndrome (CHS) and normal control cattle. Platelet aggregation induced by collagen was decreased in platelets of the cattle with CHS, but not ADP (10-20 microM), thrombin (0.5-1.0 U/ml) and phorbol-12-myristate 13-acetate (PMA, 3.2 microM). The aggregation response induced by collagen in CHS platelets lacks the change in shape which usually occurs in normal platelets. Simultaneous stimulation by collagen (10 micrograms/ml) + ADP (10 microM) is effective in restoring collagen-induced aggregating response in CHS platelet, although pretreatment of ADP (10 microM) could not restore the collagen (10 micrograms/ml)-induced aggregating response, suggesting that there is a certain threshold of stimulation intensity above which collagen-induced aggregation of CHS platelet can begin. Control normal platelets, previously exposed to ADP (10 microM) and collagen (10 micrograms/ml), showed no further response to exposure to a third aggregating agent (arachidonic acid, 5 mM). On the other hand, the final agent was able to elicit aggregating responses in CHS platelets, suggesting that the arachidonate aggregating system may be suppressed in CHS cattle, but fully activated in control animals. Furthermore, normal platelets showed a significant decrease in aggregating response to collagen when pretreated with a cyclooxygenase inhibitor, indomethacin (10(-5) M), whereas CHS platelet was insensitive to indomethacin. This indomethacin-treated normal platelet mimicked the CHS collagen-induced aggregation pattern. These data suggest that a signal transduction process from receptor-operated events to arachidonate metabolism is suppressed in collagen-induced CHS platelet aggregation.

[Significance of "overshoot" elevation of ejection fraction in early recovery period after exercise in patients with coronary artery disease].

To investigate the relationship between "overshoot" elevation of left ventricular (LV) ejection fraction (EF) in the early recovery period after exercise and myocardial ischemia, LV function was measured in 5 healthy volunteers and 47 pts with coronary artery disease (CAD) using supine ergometer. EF at rest (EFRE), at peak exercise (EFEX), at peak of overshoot (EFOS) and time from cessation of exercise to peak of overshoot (T-OS) were measured using an ambulatory LV function monitor system with a high sensitive CdTe detector. The patients were classified into 3 groups according to EF depression (EFRE-EFEX) by exercise, i.e., D1 (EF decreases less than 5%): 15 pts, D2 (EF decreases 5-10%): 12 pts, D3 (EF decreases greater than or equal to 10%): 20 pts. Degree of overshoot which shows difference in EF between rest and peak of overshoot was 23.8 +/- 2.13% (mean +/- SD) in the normal group, 11.72 +/- 5.12% in D1, 9.56 +/- 6.51 in D2 and 6.12 +/- 4.15% in D3, respectively. T-OS was 1.4 +/- 0.48 in the normal group, 2.8 +/- 0.54 in D1, 4.5 +/- 2.36 in D2 and 5.6 +/- 1.93 min in D3, respectively. We conclude that left ventricular dysfunction was proportionally related to time-lapse from cessation of exercise to peak of overshoot while inversely related to degree of overshoot.

[Usefulness of electrocardiogram gated 99mTc-methoxy isobutyl isonitrile (MIBI) single photon emission computed tomography for detection of wall motion abnormality of left ventricle].

We evaluated whether the newly developed perfusion imaging agent "99mTc-methoxy isobutyl isonitrile (MIBI)" has a possibility to detect wall motion abnormality of left ventricle. Electrocardiogram (ECG) gated single photon emission computed tomography (SPECT) was performed in 6 patients with myocardial infarction (anterior: 3 and inferior: 3). Apical and basal short axis images were selected from each patient and circumferential analysis was performed on end-diastolic (ED) and end-systolic (ES) images respectively. Count in end-diastole (EDC) and count in end-systole (ESC) were obtained, then % (ESC-EDC) and (ESC-EDC)/EDC were calculated. Wall motion of left ventricle was evaluated by either ultrasonic cardiogram or left ventriculography. Left ventricle was divided into anterior, septal, posterior and lateral areas and then each area was divided into apical and basal segments (finally the heart was divided into 8 segments). Of 48 segments, 33 segments showed normal wall motion and 15 segments showed abnormal wall motion: hypokinesis, akinesis and dyskinesis. % (ESC-EDC) was 74.39 +/- 16.85% in segment of normal wall motion and 33.27 +/- 23.56% in segment of abnormal wall motion (p < 0.001). (ESC-EDC)/EDC was 48.67 +/- 13.35% in segment of normal wall motion and 23.33 +/- 18.83% in segment of abnormal wall motion (p < 0.001). From these data, lower limit of % (ESC-EDC) and (ESC-EDC)/EDC in normal wall motion was defined as 40 and 22 respectively: mean -2SD of normal wall motion. Sensitivity of diagnosis of abnormal wall motion was 73% in % (ESC-EDC) and 60% in (ESC-EDC)/EDC (n.s.). Specificity was 94% and 97% (n.s.).(ABSTRACT TRUNCATED AT 250 WORDS)

[Property of electrocardiogram gated single photon emission tomography by 99mTc-methoxy isobutyl isonitrile].

99mTc-methoxy isobutyl isonitrile (MIBI) is a new developed myocardial perfusion imaging agent. Because this compound has higher photon energy than thallium (Tl), electrocardiogram gated single photon emission tomography (SPECT): end-diastolic (ED) and end-systolic (ES) short axis (SA) images could be taken. To investigate property of gated MIBI SPECT, MIBI myocardial scintigraphy, Tl scintigraphy (TMS) and analysis of left ventricular wall motion were performed in 6 patients with myocardial infarction. Left ventricle was divided into 8 segments. Perfusion defect (PD) was scored: "0" (normal), "1" (hypo-perfusion), "2" (defect). Wall motion abnormality (WMA) was also scored: "0" (normokinesis), "1" (hypo-kinesis), "2" (a-, dys-kinesis). Severity and extent of PD and WMA were calculated. Severity of WMA was 3.0 +/- 2.0 (M +/- SD), severity of PD was 3.3 +/- 1.7 in TMS, 3.7 +/- 1.3 in no-gated MIBI, 5.0 +/- 0.6 in ES-MIBI, 7.3 +/- 2.0 in ED-MIBI. Extent of WMA was 2.3 +/- 1.0. Extent of PD was 2.5 +/- 1.3 in TMS, 3.0 +/- 1.6 in no-gated MIBI, 3.5 +/- 0.8 in ES-MIBI, 4.8 +/- 1.0 in ED-MIBI. Compared with wall motion abnormality, severity and extent of PD in ED-MIBI was larger. From our data, it is concluded that perfusion defect in ED-MIBI was overestimated significantly. When we evaluate gated MIBI image, we must consider this property.

[Transcatheter hepatic arterial chemoembolization using microencapsulated mitomycin C (MMCmc)].

Thirty-six patients with hepatocellular carcinoma were treated by hepatic arterial chemoembolization of microencapslated mitomycin C (MMCmc). The infusion cannulae were placed in the hepatic artery by the Soldinger method, and 10 to 40 mg of MMCmc was injected 1 to 4 times (total doses, 10-110 mg). The evaluation of anti-tumor effects revealed 12 partial responses in 27 evaluable lesions and response rates were 44%. The serum AFP levels were decreased in 70% of patients. The survival rate for 12 months was 35% in all patients, but 67% at Stage III. The control of liver function is also important, because the hepatic failure and esophageal varices were the main causes of death. Side effects such as fever and pain were seen in 70%, but they were mild.

[Multi-institutional trials of radiation and Furtulon combination therapy for malignant tumors].

The efficacy of the response and safety in combination therapy of radiation and furtulon, a derivative of fluoropyrimidine, for malignant tumors were tested on a multi-institutional basis. Patients in this study were given daily 800 mg of oral furtulon and also irradiations. Twenty-three out of 30 evaluable cases showed CR or PR response (response rate was 77%). The response rates of the cases classified into regions of primary sites were 67% of stomach (4/6), 57% of colorectum (4/7), 100% of breast (9/9), 67% of esophagus (4/6), 100% of ovary (1/1) and 100% of lung (1/1). Four out of 36 cases were not given the full scheduled treatments due to grade 3 side effects, consisting of one diarrhea case suspected due to furtulon side effect, 2 impaired general condition cases according to the progression of diseases, and one case showing radiation dermatitis, dysphagia due to radiation mucositis and leukocytopenia. These results show that the combination therapy of radiation and furtulon is an efficacious and safe modality for primary and metastatic tumors.

[Experience of multimodal therapy for advanced neuroblastoma].

The prognosis of advanced neuroblastoma is extremely poor. We treated 5 patients with advanced neuroblastoma, older than 3 years, with multimodal therapy including intraoperative irradiation and autologous bone marrow transplantation. Elevated serum NSE and ferritin level and unfavorable histology according to the Shimadas histological classification, all of which are indicators of poor prognosis, were found in all of them. N-myc oncogene was amplified in 3 cases. After preoperative intensive induction chemotherapy, delayed primary operation and intraoperative irradiation (10-15 Gy) were performed. The postoperative lethal dose chemotherapy and total body irradiation (33 Gy x 3 days) were followed by autologous bone marrow transplantation. Tumor cells were purged using immunomagnetic beads method. Two cases showed recurrence (brain; 1, bone and bone marrow; 1) and a metastatic brain tumor was extirpated completely. All of them are alive during the follow up period from 6mo. to 4y8mo. (mean; 2y5mo.) with no evidence of disease except one. It may be concluded that our multimodal therapy is effective in achieving better results for advanced neuroblastoma.

[An early clinical study of Z-100 in leukopenia caused by radiotherapy].

The clinical efficacy of Z-100 for the treatment of leukopenia caused by radiotherapy for malignant tumors was evaluated in an early clinical study using three doses, 2, 10 and 20 micrograms. According to the evaluation by investigators and time-course of WBC, the highest effective rate was achieved by 20 micrograms. Adverse reactions occurred in all groups, but there were no serious or clinically relevant reactions. These results suggested that Z-100 is clinically useful for the treatment of leukopenia caused by radiotherapy.