Biomedical subjects

N Yamazaki

Publications and source records attributed to N Yamazaki.

At least 37 records · Page 2Linked to original sources

PubMed2000

Comprehensive gene expression profile of LPS-stimulated human monocytes by SAGE.

Monocytes play a pivotal role in various human infectious and inflammatory diseases. To reveal a whole picture of pathophysiologic function of activated human monocytes, this study used the serial analysis of gene expression (SAGE) procedure in lipopolysaccharide (LPS)-stimulated human monocytes. A total of 35 874 tags corresponding to more than 12 000 different transcripts were sequenced. Comparison of gene expression profile with that of resting monocytes revealed the LPS-inducible gene expression profile. Many cytokines and chemokines, including interleukin (IL)-6, IL-1alpha, IL-1beta, tumor necrosis factor (TNF)-alpha, macrophage inflammatory protein (MIP)-1beta, MIP-2beta, MIP-2alpha, liver and activation-regulated chemokine (LARC), MIP-1alpha, thymus and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC), regulated on activation, normal T cell expressed and secreted (RANTES), growth-regulated oncogene (GRO) alpha, and IL-8, were observed in the highest inducible transcripts. Other genes encoding plasminogen activator inhibitor type 2 (PAI-2), Hc-gp39, apolipoproteins, malate dehydrogenase, matrix metalloproteinase-9 (MMP-9), and cyclooxygenase (COX2) were also highly elevated in LPS-stimulated monocytes. Moreover, up-regulation of Naf1beta, IL-7 receptor, adenosine receptor A2a, and many novel genes was newly identified. These results suggest that the LPS-inducible gene products may be involved in cell activation and migration, angiogenesis, tissue remodeling, and metabolism, and thus may orchestrate the inflammatory reactions. On the other hand, the expression of numerous sets of novel genes was discovered to be down-regulated on LPS stimulation. This study represents the first comprehensive analysis of LPS-inducible gene expression in human monocytes and provides tremendous novel information for the function of LPS-activated monocytes and targets for diagnosing, monitoring, and treating sepsis and various human infectious and inflammatory diseases.

Base Sequence↗

PubMed2000

Endogenous lectins as targets for drug delivery.

To minimize side effects of drugs it would be ideal to target them exclusively to those cell types which require treatment. As a means to this end prototypical cellular recognition systems pique our interest to devise biomimetic strategies. Since oligosaccharides of glycoconjugates outmatch other information-carrying biomolecules (proteins, nucleic acids) in theoretical storage capacity by far, work on the sugar code can spark off development of effective targeting devices. Conjugation of custom-made glycan epitopes to proteins or biocompatible non-immunogenic polymeric scaffolds produces neoglycoconjugates with purpose-adaptable properties. In the interplay with endogenous receptors such as lectins, suitable oligosaccharides such as histo-blood group trisaccharides as parts of neoglycoconjugates have already proven their practical applications in histopathology. Elucidation of the structure of cell lectins with currently five main families aids to tailor ligand characteristics rationally. They include the types of functional groups and their topological presentation to optimize the bimolecular binding as well as the optimal spatial clustering and spacer characteristics to exploit cooperativity. Indeed, the potent trivalent cluster glycosides designed for the C-type asialoglycoprotein receptors furnish an instructive example how to turn the theoretical guideline on ligand modification into nM-affinity. By placing emphasis on tissue lectins as targets of neoglycoconjugate-mediated drug delivery, the long-term perspective is opened to likewise test members of these families themselves for routing of therapeutic payloads, aiming at cell addressins. This review illustrates the conceivable potential which work on the sugar code with custom-made neoglycoconjugates and tissue lectins can have in store for drug delivery.

Animals↗

PubMed2000

Specific elevation of transcript levels of particular protein subtypes induced in brown adipose tissue by cold exposure.

To understand the difference in metabolic flow in rat brown adipose tissue (BAT) from that in white adipose tissue (WAT) at the molecular level, we examined the steady-state transcript levels of 39 proteins in both adipose tissues with and without cold exposure by Northern blot analysis. In addition to the transcript levels of uncoupling protein isoforms, those of proteins involved in the transport and catabolism of fatty acids and glucose in BAT were elevated by cold exposure, suggesting the stimulation of utilization of fatty acids and glucose as fuels in BAT. As to these changes, the muscle-type subtypes were remarkable; and therefore, they were suggested to be responsible for the cold exposure-induced acceleration of energy expenditure in BAT. Furthermore, of the isoforms of beta-adrenergic receptor (beta-AR) and CCAAT enhancer binding protein (C/EBP), transcript levels of beta(1)-AR and C/EBPbeta in BAT were increased by the cold exposure. Possible roles of these proteins in energy metabolism in BAT were discussed.

Adipose Tissue, Brown↗

PubMed2000

Asymmetric synthesis of (-)-indolizidines 167B and 209D based on stereocontrolled allylation of a chiral tricyclic N-acyl-N,O-acetal.

[reaction: see text] The tricyclic N-acyl-N,O-acetal incorporating (S)-2-(1-aminoethyl)phenol as a chiral auxiliary underwent TiCl4-mediated allylation to give the chiral (5S)-allylpyrrolidinone with retention of configuration in high yield and diastereoselectivity. On the bases of this methodology, the asymmetric syntheses of the dendrobatid alkaloids (-)-indolizidines 167B and 209D were achieved.

Acetals↗

PubMed2000

Modulation of Na(+) x H(+) exchange by osmotic shock in isolated bovinearticular chondrocytes.

The effects of hyperosmotic shock on intracellular pH (pHi) have been characterized in bovine articular chondrocytes. Osmotic shock is one of a variety of physicochemical stimuli experienced by chondrocytes upon cartilage loading. Cells were isolated from their extracellular matrix, and loaded with the pH-sensitive fluorophore 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein. Hyperosmotic shocks were imposed by addition of KCl or sucrose to the extracellular medium. For cells at steady-state pHi, resuspension in hyperosmotic solutions elicited an alkalinization, which was significantly inhibited by removal of extracellular Na+ ions, or treatment with amiloride (1 mM) or HOE-694 (10 microM), both inhibitors of Na+ x H+ exchange. For cells acidified by ammonium rebound, recovery of pHi towards resting levels was significantly stimulated by exposure to hyperosmotic solutions, and the effect was again attenuated by inhibition of Na+ x H+ exchange. Determination of the rate of acid extrusion at different levels of acidification indicated that the affinity of acid extrusion systems for H+ ions was increased by hypertonic shock. The response to hyperosmotic media could be abolished by treatment of chondrocytes with the non-specific kinase inhibitor staurosporine (10 nM), while the phosphatase inhibitor okadaic acid (1 mM) was able to augment recovery rates to values similar to those measured under hyperosmotic conditions. The osmotic sensitivity of recovery was unaffected by exposure to the protein kinase C inhibitor calphostin C, but was abolished in cells treated with ML-7, a specific inhibitor of myosin light chain kinase. These results confirm that - as for other components of mechanical load - increased osmolarity can modulate the activity of Na+ x H+ exchange, in this case by altered patterns of phosphorylation of transporter-associated myosin. The changes of pHi which will result dictate in part the rate of cartilage macromolecule synthesis.

Amiloride↗

PubMed2000

Characterization of porin isoforms expressed in tumor cells.

Mitochondria from malignant tumor cell lines show a higher capability for hexokinase binding than those from normal liver. To explore possible differences in hexokinase binding sites of mitochondria between tumor cells and normal liver, we characterized porin isoforms expressed in tumor cells. Cloning experiments on the three porin isoforms, VDAC1, VDAC2 and VDAC3 from malignant tumor cell line AH130 clearly showed that their primary structures were completely identical to those of the corresponding VDACs of normal liver cells. Possible expression of the fourth porin isoform in AH130 cells was excluded by degenerate primer-based RT-PCR. However, the transcript levels of the three VDAC isoforms in AH130 cells were significantly higher than those in normal liver. These results suggest that the high hexokinase-binding capability of malignant tumor cell mitochondria was not due to any structural difference, but due to a quantitative difference in binding sites.

Alternative Splicing↗

PubMed2000

Oral administration of bovine lactoferrin for treatment of tinea pedis. A placebo-controlled, double-blind study.

A clinical study was conducted to evaluate the effectiveness of lactoferrin, which is a protein component of cow's milk, in the treatment of tinea pedis. Doses of either 600 mg or 2000 mg of lactoferrin, or a placebo was orally administered daily for 8 weeks to 37 adults who were judged to have mild or moderate tinea pedis. Dermatological improvement and antifungal efficacy were assessed. In the analysis of all subjects, dermatological symptoms scores in all groups decreased but the differences were not statistically significant comparing the three groups. However, in the analysis limited to subjects with moderate vesicular or interdigital tinea pedis, dermatological symptoms scores in the lactoferrin-treated groups decreased significantly in comparison with the placebo group (P < 0.05). The organisms isolated were Trichophyton rubrum and Trichophyton mentagrophytes. A mycological cure was not seen in any of the subjects. In the 37 subjects there were no adverse events and no subject withdrew from the study because of an adverse event. These results suggest that orally administered lactoferrin can improve the dermatological symptoms in some subjects. The potential usefulness of lactoferrin as a functional food material for treating tinea pedis was seen for the first time in this study.

Administration, Oral↗

PubMed2000

L-Arginine treatment may prevent tubulointerstitial nephropathy caused by germanium dioxide.

BACKGROUND: Long-term oral ingestion of germanium dioxide (GeO2) causes progressive renal failure derived from tubulointerstitial nephropathy in humans and animals. The characteristic of GeO2-induced nephropathy is the renal tissue injury persisting for a long time, even after cessation of GeO2 ingestion. However, a treatment that can suppress the long-lasting renal tissue injury has not yet been established. METHODS: Using the methods of immunohistochemistry and reverse transcription-polymerase chain reaction, we examined the expression of ED1-positive cells (macrophages/monocytes), transforming growth factor (TGF)-beta1 mRNA and protein and collagen type IV mRNA and protein in the kidneys of rats with GeO2-induced nephropathy. Concomitantly, the effects of L-arginine treatment on their expression was explored in the kidneys of rats with GeO2-induced nephropathy. RESULTS: Chronic administration of GeO2 caused tubulointerstitial nephropathy characterized by leukocyte invasion into the enlarged tubulointerstitial space in rats. The expression of ED1-positive cells, TGF-beta1 protein and collagen type IV protein was markedly increased in the tubulointerstitium of the renal cortex from rats with GeO2-induced nephropathy. Similarly, TGF-beta1 and collagen type IV mRNA were significantly enhanced in the renal cortex of rats with GeO2-induced nephropathy. A small number of tubulointerstitial cells expressing TGF-beta1 protein were also observed in the renal cortex of rats with GeO2-induced nephropathy. However, L-arginine treatment led to a parallel decrease in the expression of ED1-positive cells, TGF-beta1 mRNA and collagen type IV mRNA and protein in rats with GeO2-induced nephropathy. CONCLUSIONS: In general, collagen synthesis is driven by TGF-beta1 in the fibrotic process associated with a variety of renal disorders. TGF-beta1 is secreted by TGF-beta1 producing cells such as macrophages, fibroblasts and myofibroblasts. Thus, the present study indicates that the expression of collagen type IV may be mediated by TGF-beta1 released from invading macrophages and, to a lesser extent, released from tubulointerstitial cells, presumably fibroblasts and/or myofibroblasts in GeO2-induced nephropathy. L-Arginine treatment inhibits collagen type IV synthesis possibly by suppressing macrophage invasion and the resultant TGF-beta1 expression in this nephropathy. L-Arginine treatment may be beneficial in the prevention of tubulointerstitial fibrosis, which is considered to be the terminal stage of GeO2-induced nephropathy.

Animals↗

PubMed2000

Normal kinematics of the interosseous membrane during forearm pronation-supination--a three-dimensional MRI study.

We studied in vivo dynamic shape changes of the interosseous membrane (IOM) during forearm rotation using three-dimensional magnetic resonance imaging (3D-MRI), and simultaneously analysed 3D-motion of the forearm rotation. Wavy deformities were seen in the IOM in the pronated position, and similar small changes were also seen at maximum supination (average 82 degrees ) and in the neutral position. These dynamic changes mainly occurred in the membranous part of the IOM, whereas the tendinous part demonstrated minimal dynamic changes during rotation in all subjects. On the dorsal aspect, deformity around the dorsal oblique cord was seen at maximum pronation. From this 3D-MRI observation, the tendinous part is considered to be taut during rotation to provide stability between the radius and ulna, because of its straightness and less dynamic changes. The more deformable membranous part is important to allow for smooth rotation, since it lies at a distance from the rotation axis. Inelasticity developing in the membranous part from trauma may pre-dispose to pronation-supination contracture. The radius rotated around the ulna from maximum supination to 45 degrees pronation. At maximum pronation (average 75 degrees ), the radius translated average 1.8 mm palmarly and rotated average 4.0 degrees ulnarward on the ulna. Incongruity of the distal radioulnar joint, contraction of the pronator quadratus and torsion between the radius and ulna at maximum pronation may produce this irregular motion of the radius and cause the dynamic changes of the IOM.

Adult↗

PubMed2000

Structure and characterization of the genes for murine choline/ethanolamine kinase isozymes alpha and beta.

Choline/ethanolamine kinase (CK/EK) is the first enzyme in phosphatidylcholine/phosphatidylethanolamine biosynthesis in all animal cells. The highly purified CKs from mammalian sources and their recombinant gene products so far were all shown to have EK activity also, indicating that both activities reside on the same protein. CK/EK in most animal cells exists as several isoforms, for two of which (alpha and beta) their cDNAs have been cloned from both the rat and mouse, and they are found to be separate gene products. The physiological significance for the existence of more than one CK/EK enzyme, however, remains to be clarified. In this study, we isolated mouse genes encoding both types of CK/EK isozyme and determined their entire structure. The 5'-flanking promoter regions were found to have quite different features from each other, indicating that their expression could be under distinct control. Comparison of the nucleotide sequence between the corresponding coding exons showed the best homology (75%) residing on exon VIII. A search of the database resulted in the possible existence of 17 different origins of eukaryotic CK and/or EK, each of which presumably contained the entire amino acid sequence. Multialignment of their putative amino acid sequences led to an identification of the novel consensus sequence possibly required for the expression of either CK or EK activity, which corresponded to the sequence within exons VII and VIII of CK/EK-alpha and -beta genes from the mouse. This sequence was localized in close proximity to the C-terminal region of the general (Brenner's) phosphotransferase concensus sequence which was also completely conserved in all of the putative eukaryotic CK/EK proteins. The results demonstrated that, while both CK/EK-alpha and -beta genes were composed of 11 major exons, the size of their genes was quite different: 40 kb for CK/EK-alpha, whereas it was only 3.5 kb for CK/EK-beta.

Amino Acid Sequence↗

PubMed1999

Olfactory neurons expressing closely linked and homologous odorant receptor genes tend to project their axons to neighboring glomeruli on the olfactory bulb.

We have characterized two separate odorant receptor (OR) gene clusters to examine how olfactory neurons expressing closely linked and homologous OR genes project their axons to the olfactory bulb. Murine OR genes, MOR28, MOR10, and MOR83, share 75-95% similarities in the amino acid sequences and are tightly linked on chromosome 14. In situ hybridization has demonstrated that the three genes are expressed in the same zone, at the most dorsolateral and ventromedial portions of the olfactory epithelium, and are rarely expressed simultaneously in individual neurons. Furthermore, we have found that olfactory neurons expressing MOR28, MOR10, or MOR83 project their axons to very close but distinct subsets of glomeruli on the medial and lateral sides of the olfactory bulb. Similar results have been obtained with another murine OR gene cluster for A16 and MOR18 on chromosome 2, sharing 91% similarity in the amino acid sequences. These results may indicate an intriguing possibility that olfactory neurons expressing homologous OR genes within a cluster tend to converge their axons to proximal but distinct subsets of glomeruli. These lines of study will shed light on the molecular basis of topographical projection of olfactory neurons to the olfactory bulb.

Amino Acid Sequence↗

PubMed1999

Serial analysis of gene expression in human monocytes and macrophages.

Monocytes/macrophages serve as sentinels involved in chronic inflammation and the eradication of various pathogens. To define molecularly the differentiation of blood monocytes into macrophages, we conducted serial analysis of gene expression (SAGE) in human blood monocytes/macrophages induced by granulocyte-macrophage colony-stimulating factor (GM-CSF) or M-CSF. SAGE analysis of 57,560, 57,463, and 55,856 tags from monocytes, GM-CSF-, and M-CSF-induced macrophages, respectively, allowed identification of 35,037 different transcripts. Interestingly, the genes with the highest expression during differentiation from monocytes into macrophages were genes involved in lipid metabolism. Both CSF-induced macrophages expressed similar sets of genes except for several genes such as monocyte-derived chemokine (MDC), legumain, prostaglandin D synthetase, and lysosomal sialoglycoprotein. The identification of specific gene expression in human monocytes, GM-CSF-, or M-CSF-induced macrophages provides novel methods to define macrophage subsets and the maturation and activation stage of cells of macrophage lineage and, possibly, to diagnose diseases in which macrophages play a major role. This study represents the first extensive serial analysis of gene expression for any type of human hematopoietic cells.

Cells, Cultured↗

PubMed1999

Serial analysis of gene expression in human monocyte-derived dendritic cells.

Dendritic cells (DCs) are professional antigen-presenting cells in the immune system and can be generated in vitro from hematopoietic progenitor cells in the bone marrow, CD34(+) cord blood cells, precursor cells in the peripheral blood, and blood monocytes by culturing with granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4, and tumor necrosis factor-alpha. We have performed serial analysis of gene expression (SAGE) in DCs derived from human blood monocytes. A total of 58,540 tag sequences from a DC complementary DNA (cDNA) library represented more than 17,000 different genes, and these data were compared with SAGE analysis of tags from monocytes (Mo) and GM-CSF-induced macrophages (M open diamond). Many of the genes that were differentially expressed in DCs were identified as genes encoding proteins related to cell structure and cell motility. Interestingly, the highly expressed genes in DCs encode chemokines such as TARC, MDC, and MCP-4, which preferentially chemoattract Th2-type lymphocytes. Although DCs have been considered to be very heterogeneous, the identification of specific genes expressed in human Mo-derived DCs should provide candidate genes to define subsets of, the function of, and the maturation stage of DCs and possibly also to diagnose diseases in which DCs play a significant role, such as autoimmune diseases and neoplasms. This study represents the first extensive gene expression analysis in any type of DCs.

Antigen Presentation↗

PubMed1999

Expression of the bovine heart mitochondrial ADP/ATP carrier in yeast mitochondria: significantly enhanced expression by replacement of the N-terminal region of the bovine carrier by the corresponding regions of the yeast carriers.

To characterize the transport mechanism mediated by the mammalian mitochondrial ADP/ATP carrier (AAC), we tried to express bovine heart mitochondrial AAC (bhAAC) in Saccharomyces cerevisiae. The open reading frame of the bhAAC was introduced into the haploid strain WB-12, in which intrinsic AAC genes were disrupted. Growth of the transformant was very low in glycerol medium, and a little amount of bhAAC was detected in the mitochondrial membrane. For improvement of bhAAC expression in WB-12, we introduced DNA fragments encoding chimeric bhAACs, in which the N-terminal region of the bhAAC extending into the cytosol was replaced by the corresponding regions of the type 1 and type 2 yeast AAC isoforms (yAAC1 and yAAC2). These transformants grew well, and the amounts of the chimeric bhAACs in their mitochondria were as high as that of yAAC2. The carriers expressed showed essentially the same ADP transport activities as that of AAC in bovine heart mitochondria.

Amino Acid Sequence↗

PubMed1999

In vivo motion analysis of forearm rotation utilizing magnetic resonance imaging.

OBJECTIVE: To analyse in vivo normal forearm rotation utilizing the image processing method on axial magnetic resonance imaging. DESIGN: Rotatory motion and translation of the radius and rotation of the ulna were analysed in vivo, utilizing features of the bone image; axial centre of gravity and axis of the bone. METHODS: Axial magnetic resonance images at the proximal, middle and distal portion of the right forearms of 20 healthy volunteers were examined in five rotational positions, maximum pronation, 45 degrees pronation, neutral, 45 degrees supination and maximum supination. The axis of forearm rotation, the rotation angle of the radius, the rotation of the radius around its axial centre of gravity, and the rotation of the ulna around its axial centre of gravity were calculated on these axial MR images. RESULTS: The radius rotates in a near-circular ellipse around its rotational centre located on a line through the radial head and the ulnar head from maximum supination to 45 degrees pronation. There was discrepancy between the calculated maximum rotational angle of the radius and the pronation-supination angle showed on the goniometer. This discrepancy was due to motion at the radiocarpal, midcarpal and carpometacarpal joints. The rotation of the ulna ranges within 6 degrees. At maximum pronation, there was palmar translation of the radius. CONCLUSIONS: The forearm rotation is not a simple rotatory motion but a complex motion with rotation and translation. The palmar translation of the radius at maximum pronation may be caused by incongruity of the distal radioulnar joint and contraction of the pronator quadratus muscle. RELEVANCE: To date there exist few methods for the accurate and non-invasive assessment for motion analysis of the forearm rotation. Such a technique utilizing magnetic resonance image characteristics is, however, helpful to reveal accurate self- and relative-rotatory motion of both radius and ulna as well as the translation movement on forearm rotation in normal forearm.

Adolescent↗

PubMed1999

Three-dimensional magnetic resonance imaging of the interosseous membrane of forearm: a new method using fuzzy reasoning.

We now report newly developed three-dimensional magnetic resonance imaging (3D-MRI) system which is based on semiautomatic tissue extraction from the axial MR images utilizing the fuzzy reasoning calculation method and 3D-image reconstruction with surface rendering. We also studied normal in vivo dynamic changes of the interosseous membrane (IOM) of forearm during rotation using this 3D-MRI. Serial axial MRI of right forearms of five healthy volunteers was obtained in five rotational positions, and extraction and 3D-reconstruction of the radius, ulna, and IOM was made using the system. Extraction results were well with the fuzzy reasoning method. 3D-MRI of the radius and ulna, IOM were reconstructed from these images respectively, and their 3D-shapes were almost identical to the anatomic shape. 3D-MRI showed there were wavy deformities on the IOM in pronation position in the all five subjects and dorsiflexion on the most dorsal portion of the IOM at maximum supination in three forearms. In neutral position, the IOM of all five volunteers was almost flat. From anatomic orientation, these dynamic changes of the IOM mainly occurred at the membranous portion, which is soft, thin, and elastic. Otherwise, the tendinous portion which is a thick and strong complex of 5 to 10 bundles run from proximal one third of the radius to distal one fourth of the ulna, demonstrated minimal dynamic changes on the 3D-MRI. Therefore, the tendinous portion is considered to be taut during rotation to provide stability between the radius and the ulna, while the membranous portion is easy to deform and allowing smooth rotation. Furthermore, because of wide-use, our 3D-MRI system is useful for in vivo analysis of soft tissue kinesiology in normal and abnormal musculoskeletal systems.

Adult↗

PubMed1999

Low frequency of neutralizing antibodies against natural interferon-beta during adjuvant therapy for Japanese patients with melanoma.

The incidence of therapy-induced neutralizing interferon-beta (IFN-beta) antibodies was investigated in 41 Japanese patients with melanoma treated with a combination of chemotherapy and subcutaneous injections of natural IFN-beta. Only one patient (2.4%) developed anti-IFN-beta antibodies. This finding differs from those of previous studies with Caucasian patients in which subcutaneous injections of natural IFN-beta resulted in a high incidence of anti-IFN-beta antibodies. This difference may be related to ethnic group of patients, treatment schedule, pharmacological preparation of IFN-beta and/or technical assay system used in each study.

Adult↗

PubMed1999

Dacarbazine, nimustine hydrochloride, cisplatin and tamoxifen combination chemotherapy for advanced malignant melanoma.

Melanoma is an uncommon disease in Japan. The incidence, however, has been gradually increasing in the last two decades, as in many other countries worldwide. Ten patients with metastatic malignant melanoma were treated between March of 1997 and April of 1998 in the Department of Dermatology, National Cancer Center Hospital, with a combination chemotherapy consisting of dacarbazine (DTIC), nimustine hydrochloride (ACNU), cisplatin (CDDP), and tamoxifen (TAM). The patients characteristics were as follows: four were males and six females; the age range was 33-70 years; all were Japanese; sites of primary disease: extremities 4, primary unknown 3, nasal cavity 1, anus 1, scalp 1; sites of metastases: lymph nodes 6, pulmonary system 5, skin 2, liver 3, gall bladder 1, adrenal gland 1. The chemotherapy regimen included DTIC 220 mg/m2/i.v. on days 1 through 3, ACNU 60 mg/m2/i.v. on day 1, cisplatin 25 mg/m2/i.v. on days 1 through 3, and tamoxifen 10 mg p.o. twice daily. One patient achieved a complete response and 3 showed partial responses. The response rate was 40%. The four responders included those with metastases to the nodes, lung, and liver. The main toxicities were nausea, vomiting, leucopenia, anemia, and thrombocytopenia. This regimen is a fairly effective combination against metastatic melanoma.

Adult↗