Tetsuo Maita

Resveratrol Induces Downregulation in Survivin Expression and Apoptosis in HTLV-1-Infected Cell Lines: A Prospective Agent for Adult T Cell Leukemia Chemotherapy

Abstract: Resveratrol, a phytoalexin found in grapes and wine, has been shown to exhibit a wide range of pharmacological properties and is believed to play a role in the chemoprevention of human cancer. Resveratrol has also been shown to induce antiproliferation and apoptosis of several leukemia cell lines. In the present study, we investigated the effect of resveratrol in adult T cell leukemia. Our present observations showed that resveratrol induced growth inhibition in all five human T cell lymphotrophic virus-1-infected cell lines examined, with 50% effective dose of 10.4-85.6 mM. In the resveratrol-treated cells, induction of apoptosis was confirmed by annexin V-based analyses and morphological changes. The most surprising observation was that resveratrol treatment resulted in a gradual decrease in the expression of survivin, an antiapoptotic protein, during cell apoptosis. These findings indicate that resveratrol inhibits the growth of human T cell lymphotrophic virus-1-infected cell lines, at least in part, by inducing apoptosis mediated by downregulation in survivin expression. In view of the accumulating evidence that survivin may be an important determinant of a clinical response in adult T cell leukemia, our present findings have led to the suggestion that resveratrol, a common constituent of the human diet, merits further investigation as a potential therapeutic agent for this incurable disease.

The primary structure of L-1 light chain of chicken fast skeletal muscle myosin and its genetic implication

It is known that in general fast skeletal muscle myosin consists of 2 heavy chains and 4 light chains [ 1,2]. We have also recognized 4 components in the light chain fraction from chicken fast skeletal muscle myosin by CelIulogel electrophoresis at pH 8.3, and designated these light chains L-l, L-2, L-3 and L-4 light chains in the order of their increasing anionic mobilities [3]. L-l is also known as alkali light chain 1 (Al) and L-4 as alkali light chain 2 (A2). L-2 and L-3 are also called DTNB light chains. L-3 is a phosphorylated L-2 light chain. The primary structures of L-2 and L-4 light chains of chicken fast skeletal muscle myosin have been reported in [4,.5]. We describe here the primary structure of the L-l light chain from chicken fast skeletal muscle myosin and its genetic implication.

The L-2 light chain of chicken skeletal muscle myosin.

As with rabbit skeletal muscle myosin, four kinds of light chains may be separated from chicken skeletal muscle myosin by cellogel electrophoresis at pH 8.3. They are designated L-l, L-2, L-3 and L-4 in the order of their increasing anionic mobilities. L-1 is also known as alkali light chain 1 (A1) and L-4 as alkali light chain 2 (A2). L-2 and L-3 are also called DTNB light chain. L-3 is a phosphorylated L-2 light chain. Interest in the role of these myosin light chains in the mechanism of muscle contraction has been marked. The primary structures of L-1 and L-4 light chains of rabbit skeletal muscle myosin have been determined by Frank and Weeds [1] ; they contain 190 and 149 amino acids, respectively. The primary structure of the L-2 light chain of rabbit skeletal muscle myosin has been determined by Collins [2] and by Matsuda et al. [3]. The amino acid composition of the L-2 light chain of chicken skeletal muscle myosin has been found by Lowey and Holt [4] and its partial amino acid sequence reported by Jakes et al. [5]. We report here the primary structure of the L-2 light chain of chicken skeletal muscle myosin.

Amino acid sequences of the cardiac L-2A, L-2B and gizzard 17 000-Mr light chains of chicken muscle myosin.

Myosins separated from various vertebrate muscles contain 2 heavy chains of -200 000-M, and 4 light chains of -20 OOOllcl, [l-4]. In connection with a relationship between the light chain structure and mechanism of the muscle contraction, we have studied the primary structures of the light chains of the fast skeletal, cardiac and gizzard muscle myosins from chicken [5-l 11. Chicken fast skeletal muscle myosin has 4 kinds of light chains designated L-l-L-4 in order of their increasing anionic mobilities at pH 8.3 [5,6,8-lo]. The L-l and L-4 light chains are also called alkali light chain 1 (Al) and alkali light chain 2 (A2), respectively. The L-2 and L-3 light chains are also known as DTNB light chains. L-3 is a phosphorylated L-2 light chain [l-3]. The primary structures of these light chains have been reported [5,6,8-lo]. Chicken cardiac muscle myosin has 2 kinds of light chains designated L-l and L-2, and the primary structure of the L-l light chain has been already reported [7,8]. We have also recognized 2 components in the L-2 light chain fraction by cellulogel electrophoresis at pH 8.3 after performic acid oxidation, and designated these components L-2A and L-2B. Chicken gizzard muscle myosin contains 2 kinds of light chains. One of them is called 20 000-M, light chain or GI light chain. It is also called regulatory light chain because it has calcium binding ability [4]. The other is called 17 000-M, light chain or GII light chain [4]. The primary structure of the 20 000-M, light chain has been reported [ 111. Here, we present the primary structures of the

Matrix metalloproteinase-9 and vascular endothelial growth factor : a possible link in adult T-cell leukaemia cell invasion

Summary. Plasma from a total of 57 patients with adult T‐cell leukaemia (ATL) (acute ATL, 39 patients; lymphoma ATL, one patient; chronic ATL, 15 patients; smouldering ATL, two patients) and 20 healthy controls was analysed for the presence of type IV gelatinase activity with clinical features. A significant elevation of plasma matrix metalloproteinase‐9 (MMP‐9) was observed in some ATL patients, particularly in the patients with malignant cell infiltration. MMP‐9 was found to be secreted into the conditioned medium from all ATL cell lines examined. Moreover, the corresponding mRNA was detectable both in all ATL cell lines examined and in the majority of primary acute ATL cells, indicating that ATL cells are capable of synthesizing and secreting MMP‐9. We previously demonstrated that a high incidence of ATL cell infiltration was closely related to a high plasma level of vascular endothelial growth factor (VEGF) produced by ATL cells themselves. This present study showed that the presence of increased plasma MMP‐9 was closely associated with elevated plasma VEGF in ATL patients. Furthermore, we showed that both increased plasma MMP‐9 and VEGF were significantly related to high ATL cell infiltration. All these findings strongly suggest that MMP‐9 and VEGF act co‐operatively in the process of ATL cell invasion.

The Amino Acid Sequences of the Two Main Components of Adult Hemoglobin from Orangutan (Pongo pygmaeus)

Zusammenfassung: Die beiden Hauptkomponenten des Hämoglobins aus Orang-Utan-Blut wurden, ohne sie vorher zu trennen, vom Häm befreit; die Globine wurden dann durch Säulenchromatographie an CM-Cellulose getrennt. Dabei erhielt man zwei verschiedene erKetten (a-I, α-II) und eine ß-Kette. Die tryptischen Peptide der einzelnen 5-aminoethylierten Ketten wurden isoliert und sequenziert. Die tryptischen Peptide wurden in Homologie zur Primärstruktur des adulten Humanhämoglobins angeordnet, wodurch sich die Primärstrukturen der einzelnen Ketten ableiten ließen. Im Vergleich zum Humanhämoglobin liegen bei der a-IKette drei, bei der a-II-Kette und bei der 0-Kette je zwei Aminosäureaustausche vor; außerdem unterscheidet sich die α-1-Kette von der α-11-Kette nur durch den Austausch von Asparaginsäure gegen Glycin in Position 57.

The amino acid compositions of the tryptic, chymotryptic and peptic peptides from the L-2 light chain of rabbit skeletal muscle myosin.

The light chain fraction was separated from rabbit skeletal muscle myosin and four kinds of light chains, L-1, L-2, L-3 and L-4 in the fraction were further isolated by column chromatography using DEAE-cellulose DE-52. After amino-ethylation, the L-2 light chain was digested with trypsin. It was also digested with chymotrypsin and pepsin, respectively, after carboxymethylation. Each of the tryptic, chymotryptic and peptic peptides thus obtained was separated and purified and their amino acid compositions were analyzed.

Structure and function of muscle myosin

We have studied the primary structures of myosins from chicken muscles in order to clarify the relationship between structure and function of muscle myosin. The primary structures of the various kinds of light chains from chicken muscle myosins have been determined. We also report the primary structure of the 23K fragment of subfragment-1 (S-1) component from the heavy chain of chicken fast skeletal muscle myosin. In addition, antibody was prepared against the 23K fragment. The antibody was found to inhibit the Mg2+-ATPase activity and the initial Pi burst of the ATPase in the S-1 component. The antibody suppressed the ATP-induced fluorescence enhancement of S-1, though it did not suppress the binding of ATP to S-1. These results are also discussed.