Potent and broad antitumor effects of DX-8951f, a water-soluble camptothecin derivative, against various human tumors xenografted in nude mice.

PURPOSE: We have previously reported that DX-8951f, a water-soluble and nonprodrug camptothecin (CPT) derivative, exhibits both high in vitro potency against a series of 32 malignant cell lines and significant topoisomerase I inhibition. The purpose of this study was to evaluate the therapeutic efficacy of DX-8951f against human tumor xenografts in nude mice and to compare its activity with those of CPT-11 and other current CPT derivatives. METHODS: The antitumor activity of DX-8951f against xenografts of several different types of human tumors was determined in nude mice using a schedule in which DX-8951f was administered intravenously every 4th day for a total of four injections. RESULTS: Against both gastric adenocarcinoma SC-6 and its CPT-11-resistant variant, SC-6/CPT-11, DX-8951f demonstrated superior antitumor activity and antitumor activity over a broader range of doses than did CPT-11, SK&F104864 (hycamtin, topotecan) and GG-211 (GI147211). DX-8951f at 75 mg/kg was effective (growth inhibition rate IR > or = 58%) against 15 of 16 lines of human cancers examined (6 colon cancers, 5 lung cancers, 2 breast cancers, 1 renal cancer and the above 2 gastric cancers), and exhibited excellent antitumor activity (IR > or = 80%) against 14 of these lines. CPT-11 exhibited antitumor activity with IR values of 58% and higher against 11 lines and IR values of 80% and higher against only eight of the same 16 human tumors. DX-8951f was effective in inhibiting the growth of an SN-38-resistant tumor and some P-glycoprotein-expressing tumors, but CPT-11 was not. CONCLUSIONS: DX-8951f exhibited potent antitumor activity against various types of human tumor xenografts. Its in vivo antitumor effects were superior to those of current camptothecin analogs against certain tumors.

Role of excision repair in UVB-induced depletion and recovery of human epidermal Langerhans cells.

Skin exposure to UVB radiation deprives the antigen-presenting function and OKT6 monoclonal antibody-binding characteristics of Langerhans cells. Decrease of Langerhans cell population could be relevant to immune surveillance disturbance in the UV-exposed skin. Patients with xeroderma pigmentosum exhibit a thousandfold higher risk for sunlight-induced skin cancers than patients with normal skin, and also have various defects in cellular immunity. Therefore, studies of numerical and structural changes in epidermal Langerhans cells of patients with xeroderma pigmentosum after UVB irradiation compared with normal subjects may contribute to understanding the role of antigen-presenting cells in photocarcinogenesis. The effect of UVB radiation on OKT6+ Langerhans cells was studied in epidermal sheets obtained from irradiated normal subjects (36 and 49 years old) and subjects with xeroderma pigmentosum complementation group A (21 years old), complementation group D (32 years old), and variant (35 and 60 years old). Langerhans cell densities in chronically sunlight-exposed skin were remarkably reduced in patients with xeroderma pigmentosum group A but only slightly in those with xeroderma pigmentosum variant and in normal subjects compared with covered skin. Structural changes were substantial in Langerhans cells of chronically exposed patients with xeroderma pigmentosum group A but fewer in subjects with other xeroderma pigmentosum groups and in normal subjects. A single irradiation of three times the minimal erythema dose induced a large reduction of Langerhans cells from 3 to 7 days in all subjects. However, subsequent reappearance and return to preirradiated levels was delayed more in patients with xeroderma pigmentosum group A than in those with xeroderma pigmentosum variant and normal subjects. These results indicate (1) an essential role for excisional repair in the UVB-induced depletion, recovery, and maintenance of the epidermal Langerhans cell population, and (2) a possible, but not confirmed, relationship between depletion of Langerhans cells and earlier photocarcinogenesis in xeroderma pigmentosum.

Multicentric reticulohistiocytosis. Electron microscopic and ultracytochemical studies.

Lesions from a patient affected with multicentric reticulohistiocytosis (MR) were studied by ultracytochemical techniques to determine the relationship of PAS-positive materials and/or acid phosphatase (AP) activity to electron-dense granules found in the large mononucleated or multinucleated giant cells that are characteristic of this entity. The lead phosphate reaction products that mark the sites of AP activity were found in the electron-dense granules. The granules were also PAS positive by the periodic acid-thiosemicarbazide-silver proteinate method. These findings suggest that the PAS-positive and diastase-resistant materials in the large mononucleated or multinucleated giant cells of MR are not phagocytized materials but are pre-existing lysosomes.

Systemic administration of a synthetic lipid A derivative, DT-5461a, reduces tumor blood flow through endogenous TNF production in hepatic cancer model of VX2 carcinoma in rabbits.

Intravenous administration of DT-5461a, synthetic low-toxicity lipid A derivative, significantly inhibited the growth of VX2 tumor transplanted in the liver of rabbits. DT-5461a induced high levels of TNF activity in tumor tissue from 30 to 60 min after the administration, while no TNF activity was detected in the adjacent nontumorous liver tissue. Simultaneous measurement of microcirculatory blood flow in both tumorous and nontumorous regions in the liver by laser doppler velocimetory revealed that intravenous administration of DT-5461a caused a significant blood flow reduction in tumor region, but not in nontumorous counterparts. Tumor blood flow was significantly reduced by 40 to 60% at 30 to 90 min after the DT-5461a administration as compared with preadministration value. In contrast, local administration of human recombinant TNF alpha through the hepatic artery induced blood flow reduction not only in tumor region but also in nontumorous liver tissue. These results suggest that systemic administration of DT-5461a induced selective tumor microcirculatory blood flow reduction via local endogenous TNF production.

DT-5461a, an antitumor synthetic lipid a analog, causes selective blood flow reduction in tumor tissue.

We previously reported that a synthetic low-toxicity lipid A analog, DT-5461a, exhibited a significant antitumor effect was characteristically accompanied by extensive tumor necrosis, suggesting that DT-5461a causes a local circulatory disturbance in tumor tissues. In this study, we investigated the effect of DT-5461a on regional blood flow in various organs including tumor tissue with a radiolabeled tracer-distribution technique using 14C-iodoantipyrine. Intravenous administration of DT-5461a induced blood flow reduction in Meth A tumor subcutaneously implanted into BALB/c mice, but not in liver, spleen or lung of these mice. This tumor tissue-specific reduction in blood flow was significantly inhibited by pretreatment with antisera against tumor necrosis factor (TNF) alpha, interferon (IFN) alpha/beta, and IFN gamma. These results indicate that endogenously induced cytokines, namely TNF alpha and IFNs, are involved in the intratumor blood flow reduction caused by DT-5461a.

Molecular cloning and identification of bottle-nosed dolphin p40(phox), p47(phox) and p67(phox).

The bottle-nosed dolphin NADPH oxidase cytosolic components, p40(phox), p47(phox) and p67(phox) cDNA's were cloned from mitogen stimulated peripheral white blood cell mRNA utilizing the reverse transcription-polymerase chain reaction. The sequences of these cDNAs showed that dolphin p40(phox), p47(phox) and p67(phox) clones contained open reading frames encoding predicted polypeptides of 339, 391 and 526 amino acids, respectively. Analysis of the p47(phox) and p67(phox) amino acid sequences showed two potential Src homology three domains and p40(phox) one. Comparison of the deduced amino acids showed that dolphin p40(phox) sequence shared 88.8% similarity with the human p40(phox), that dolphin p47(phox) sequence shared 87.7% similarity with the bovine p47(phox), and that dolphin p67(phox) shared 88.1% similarity with the bovine p67(phox). Western blot analysis using anti-human p40(phox), p47(phox) and p67(phox) antibodies demonstrated that dolphin neutrophil possesses p40(phox), p47(phox) and p67(phox) with similar molecular masses and structures, to each counterpart in human neutrophils, except for the p67(phox) COOH-terminus. These results suggest that dolphin NADPH oxidase cytosolic components have functional activities equivalent to those of human.

Molecular cloning and functional expression of bottle-nosed dolphin (Tursiops truncatus) interleukin-1 receptor antagonist.

The bottle-nosed dolphin (Tursiops truncatus) interleukin-1 receptor antagonist IL-1ra cDNA was cloned from mitogen-stimulated peripheral blood mononuclear cells (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The sequence of this cDNA showed that dolphin IL-1ra clones contained open reading frames encoding 177 amino acids. Comparison of the deduced amino acids showed that dolphin IL-1ra sequence shared 87.6, 77.9, 77.4, 77.4, 76.4, and 75.8% similarity with the bovine, rabbit, equine, human, mouse, and rat IL-1ra sequences, respectively. Recombinant glutathione S-transferase (GST) dolphin IL-1ra produced in Escherichia coli (E. coli) was purified. This protein suppressed the cytostatic activity of dolphin IL-1beta on A375S2 cells, indicating that the dolphin IL-1ra cDNA obtained in the present study encodes biologically active dolphin IL-1ra.

Molecular cloning and identification of bottle-nosed dolphin flavocytochrome b gp91(phox) and p22(phox) subunits.

The bottle-nosed dolphin (Tursiops truncatus) gp91(phox) and p22(phox) cDNA were cloned from mitogen stimulated leukocytes RNA utilizing the reverse transcription-polymerase chain reaction. The sequences of these cDNAs showed that dolphin gp91(phox) and p22(phox) clones contained open reading frames encoding 569 and 192 amino acids, respectively. Analysis of the gp91(phox) amino acids sequence showed three potential N-linked glycosylation sites. Comparison of the deduced amino acid showed that dolphin gp91(phox) sequence shared 95.4, 93.8, 91.4 and 89.5% similarity with the bovine, porcine, human and mouse gp91(phox) sequences, respectively. Similarly, the amino acid sequence showed that dolphin p22(phox) shared 89.7, 84.6, 84.1, 83.6 and 83.6% similarity with the bovine, mouse, porcine, human and rattus p22(phox) sequences, respectively. Western blotting analysis with anti-peptide antibodies supported the molecular weights of the dolphin gp91(phox) and p22(phox) homologous proteins predicted from the cDNAs and amino acids sequence data.

Antenatal diagnosis of fetal abnormality by ultrasonic real time scanner.

Recent progress in antenatal diagnosis has made it possible to detect most fetal malformations which can be treated and cured by neonatal surgery. In this study an analysis of antenatal diagnosis by ultrasonography in the author's clinic is reported.

Thermal stability of nanocrystalline diamond films grown by biased enhanced microwave plasma chemical vapor deposition.

The thermal stability of nanocrystalline diamond (NCD) films grown on mirror-polished silicon substrates by biased enhanced microwave plasma chemical vapor deposition was investigated. Different pieces of a NCD sample were annealed for 1 h in an ambient argon atmosphere at 200, 400, 600, and 800 degrees C. The structural and mechanical properties of as-grown and annealed samples were assessed. The surface roughness and high hardness of the samples remained fairly constant with annealing temperature.

[Amplification of L-myc oncogene in malignant meningioma. Case report].

Amplification of L-myc oncogene was noticed in a malignant meningioma originating from the right sphenoidal wing of a 54-year-old female. The patient underwent three surgical resections plus radiotherapy over a period of 11 years and then the growth rate of the tumor became much greater with a severely invasive appearance. Using Southern blot hybridization, L-myc amplification was examined on the specimen resected at the fourth operation. As a result, approximately five-fold amplification was confirmed, which has not been previously reported except for that in a small cell carcinoma of the lung. This result may suggest that L-myc amplification is responsible to some extent for the malignant transformation in this meningioma.

[Combination therapy with granulocyte colony-stimulating factor (G-CSF) and erythropoietin (EPO) induced prominent granulocyte increase in an elderly case of myelodysplastic syndrome (MDS)].

A 70-year-old man was admitted to our hospital with pancytopenia. He was diagnosed as having MDS (RA), and therapy with subcutaneous S-CSF (100 micrograms/day) was started. His leukocyte count increased from 800/microliters to 1,400/microliters in two weeks. The dose of G-CSF was raised to 200 micrograms/day in the third week, and leukocytes increased to 2,00/microliters. At the fifth week, intravenous EPO (6,000 U x 3 times/week) was added. His leukocyte count increased to 4,000/microliters. EPO therapy was raised to 12,000 U x 3 times/week at the eighth week, his leukocyte count remained at the same level. G-CSF and EPO was stopped at the eleventh week, and leukocytes decreased to the same level as before administration. Throughout the course, his platelet count and reticulocyte count did not change. G-CSF and EPO are known as the stimulators of granuriod and erythroid progenitor, respectively. However, in this case, combination therapy with G-CSF and EPO induced marked increase of granulocytes only. This was an interesting case in relation to the roles of these cytokines in the hematopoietic system.

[Binding characteristics of RU 486 with glucocorticoid and progestin receptors].

RU 34886 (RU 486) has been proved to fully antagonize the actions of glucocorticoid and progestin at the receptor level. The binding characteristics of RU 486 with the thymic glucocorticoid receptor (GR) and the uterine progestin receptor (PR) were investigated in order to elucidate the mechanism of these antagonistic actions. The ability of RU 486 was studied to promote the "activation" and the "nuclear translocation" of GR and RP. Under heat activation, RU 486 dissociated faster from the activated GR than the non activated, and the binding complex of RU 486 and GR showed lower affinity for DNA-cellulose than the glucocorticoid agonist-GR complex. Nearly undetectable amounts of RU 486 were recovered in the nucleus. Conversely, the RU 486-PR complex showed no difference of dissociation rate between the activated and the non activated condition. Its affinity for DNA-cellulose was the same as that of the activated progestin agonist-PR complex. A large amount of this compound was demonstrated in the nucleus.

PIP: RU 38486 (RU 486) has been proven to fully antagonize the actions of glucocorticoid and progestin at the receptor level. The binding characteristics of RU 486 along with the thymic glucocorticoid receptor (GR) and the uterine progestin receptor (PR) were investigated in order to clarify the mechanism of these antagonistic actions. The ability of RU 486 was studied in order to promote the activation and the nuclear translocation of GR and PR. Under heat activation, RU 486 dissociated faster from activated GR than the nonactivated and the binding complex of RU 486 and GR showed lower affinity for DNA-cellulose than the glucocorticoid agonist-GR complex. Nearly undetectable amounts of RU 486 were recovered in the nucleus. Conversely, the RU 486-PR complex showed no difference of dissociation rate between the activated and the nonactivated condition. Its affinity for DNA-cellulose was the same as that of the activated progestin agonist-PR complex. A large amount of this compound was demonstrated in the nucleus. (author's)

Comparison of plasma prostaglandins in the non-pregnant and pregnant women: a longitudinal study.

Plasma levels of PGE2 and PGF2 alpha were determined in women from 24 to 40 weeks of pregnancy with non-pregnant women as controls. The plasma levels of PGE2 showed no significant changes as gestation advanced except for late pregnancy only. Furthermore, there were no significant differences observed between the control and pregnant subjects except late in pregnancy. On the other hand, the concentrations of PGF2 alpha in the pregnant subjects revealed a significant increase (p less than 0.005) compared with those of the controls, though no significant changes were observed with the advance of gestation except for late pregnancy. The PGF2 alpha/PGE2 ratio showed a six to ten times rise during pregnancy (p less than 0.005) compared with that of the controls. Therefore, the results obtained in the present study may explain the effectiveness of prostaglandin synthetase inhibitors in suppressing preterm uterine contraction and they also indicate the involvement of some factors in the initiation and inhibition of the myometrium contraction.

[Quantitative studies on maternal perception of fetal movement with two ultrasonic real-time scanners].

In this study, the relationship between actual fetal movement in utero and maternal perception was investigated with the use of two ultrasonic real-time scanners and a cardiotocogram. The movements of body, arms, legs, fetal breathing, uterine contractions and maternal perception of fetal movement were quantitatively analyzed. 37% of all fetal movements were recognized by the mother. Only 15% of isolated movements of fetal extremities were perceived by the mother. 72% of strong body movements with both hands and legs were recognized. No relationship between the duration and times of fetal movement and gestational weeks was found. On the other hand, a close relationship between the maternal perception rate and gestational weeks was found (28-30W, 48%; 34-35W, 53%; 40W, 67%; R = 0.903). As for the parity of the mother, multiparous women were found to be more sensitive than primiparous women (p less than 0.005). (This study was supported by Alexander von Humboldt-Stiftung in F.R.G.).

[Do OKT6 positive Langerhans cells play a role in the suppression of ultraviolet-induced carcinogenesis?].

Langerhans cells (LC) in epidermis are antigen presenting cells. LC may play a role in immune surveillance system and are considered to suppress development of ultraviolet (UV) induced skin cancers. We studied effect of UVB irradiation to LC of xeroderma pigmentosum (XP) and normal subjects by using OKT6 monoclonal antibody. When 3 minimal erythema dose (MED) of UVB were irradiated, density of OKT6 positive LC of XP began to decrease 6 hours after irradiation, and showed the least numbers on day 2 and returned completely to the pre-irradiation level on day 14. Further, after 3 MED irradiation, LCs of both normal subjects became the least on day 3 and returned to the pre-irradiation level on day 14. In XP variant and normal subjects, the number of LC in chronic sun-exposed skin decreased significantly in a similar way comparing to that of non-exposed skin. These results suggest that epidermal LC may not play an essential role in prevention of UV-induced tumor development.

Effect of combined administration of a synthetic low-toxicity lipid A derivative, DT-5461a, and indomethacin in various experimental tumor models of colon 26 carcinoma in mice.

We investigated the antitumor effects of a synthetic lipid A derivative, DT-5461a, in combination with indomethacin in three experimental tumor models (peritoneal carcinomatosis, liver tumor, and lung tumor models) of transplanted colon 26 carcinoma in mice. This carcinoma produces the immunosuppressive prostaglandin E2 (PGE2). Intravenous administration of DT-5461a alone resulted in little or no prolongation of survival time [increase in life span (ILS): -2%-22%]. When indomethacin was given in drinking water a slight or moderate increase in survival time was seen (ILS: 4%-45%). In contrast, the combination of DT-5461a and indomethacin induced an additive increase in life span (ILS: 16% to more than 193%). The strongest antitumor effect of this combined therapy was seen in the peritoneal carcinomatosis model; in this model, plasma PGE2 concentrations were considerably higher than in normal mice, and concentrations were further but transiently increased by DT-5461a administration. Following oral indomethacin administration, these elevated PGE2 concentrations were reduced to the level in untreated normal mice. Furthermore, intratumoral tumor necrosis factor (TNF) activity in the group receiving the combined therapy was significantly higher than that in the DT-5461a-treated group. No TNF production was induced by the administration of indomethacin alone. These results suggest that the antitumor effect of DT-5461a can be enhanced by combination with indomethacin, and that the inhibition of PGE2 production may have a role in this antitumor effect.

[Study on the histopathological abnormalities of the umbilical cord].

There have been very few reports of studies on histological abnormalities of the umbilical cord. In the present study, we have investigated various abnormalities of the cord with special emphasis on the cord vasculature and the vestigial remnants based on developmental abberations. The incidence of single umbilical artery was 1/509 (0.2%), by which no special relation was signified between the single artery and the occurrence of congenital malformations of the neonate. Supernumerary umbilical vessels were found in 5/509 (1.0%) of the cases examined. Three out of five revealed the umbilical cord with two arteries and two veins, while two of them showed the umbilical cord with the normal 3 vessels associated with an accessory fourth vessel. It is of a great interest to note that fetal distress with or without fetal acidosis was demonstrated in all cases with the accessory fourth vessel, whereas the presence of supernumrary umbilical vessels was least related to the occurrence of congenital malformations of the neonates. The incidence of vestigial remnants was 30/509 (5.9%). The vestigial remnants were classified into two groups according to their epithelial pattern, i.e. flat epithelium and cuboidal or columnar epithelium. The vestigial remnants with flat epithelium type, considered to originate in the allantoic duct, were found in 2/3 (66.7%) of all cases with vestigial remnants. The vestigial remnants with cuboidal or columnar epithelium, considered to originate in the omphalomesenteric duct, were found in 1/3 (33.3%) of all cases with vestigial remnants, one of which case was associated with congenital malformation of omphalocele of the neonate.

The development of a computer-aided perinatal information system.

In the perinatal center of Kagawa Medical School, we have newly developed and introduced a computer-aided perinatal information system to realize an ideal management system for high-risk pregnancies. The system comprises 5 microcomputer subsystems. With this system, not only can the continuous full monitoring of the fetus during labor and delivery be easily realized, but also can the management of outpatients and perinatal statistics. This system also supports the telephone line network. Further, we have developed a highly convenient portable CTG that can easily be operated by the patient at home. Through the use of this portable CTG and the computer system, the information about fetal heart rate and uterine contractions of high-risk pregnancy can be easily and directly transmitted to the hospital from the patient's home. This system is indispensable to reducing the perinatal morbidity and mortality.