BAALC potentiates oncogenic ERK pathway through interactions with MEKK1 and KLF4.

Although high brain and acute leukemia, cytoplasmic (BAALC) expression is a well-characterized poor prognostic factor in acute myeloid leukemia (AML), neither the exact mechanisms by which BAALC drives leukemogenesis and drug resistance nor therapeutic approaches against BAALC-high AML have been properly elucidated. In this study, we found that BAALC induced cell-cycle progression of leukemia cells by sustaining extracellular signal-regulated kinase (ERK) activity through an interaction with a scaffold protein MEK kinase-1 (MEKK1), which inhibits the interaction between ERK and MAP kinase phosphatase 3 (MKP3/DUSP6). BAALC conferred chemoresistance in AML cells by upregulating ATP-binding cassette proteins in an ERK-dependent manner, which can be therapeutically targeted by MEK inhibitor. We also demonstrated that BAALC blocks ERK-mediated monocytic differentiation of AML cells by trapping Krüppel-like factor 4 (KLF4) in the cytoplasm and inhibiting its function in the nucleus. Consequently, MEK inhibition therapy synergizes with KLF4 induction and is highly effective against BAALC-high AML cells both in vitro and in vivo. Our data provide a molecular basis for the role of BAALC in regulating proliferation and differentiation of AML cells and highlight the unique dual function of BAALC as an attractive therapeutic target against BAALC-high AML.

Voriconazole is effective as secondary antifungal prophylaxis in leukemia patients with prior pulmonary fungal disease: case series and review of literature.

Although previous invasive fungal diseases (IFD) pose a significant risk of reactivation during successive chemotherapies in patients with acute leukemia, and secondary antifungal prophylaxis is regarded as mandatory, much remains unknown about the optimal strategy including the efficacy of voriconazole. During January 2006 and October 2008, a total of 15 patients with acute leukemia had pulmonary IFD (4 probable and 11 possible cases) before or during chemotherapy. After successful treatment of primary IFD with oral voriconazole, all of them received voriconazole during a total of 35 courses of successive chemotherapy. All but one patient successfully accomplished planned treatment without suspected IFD or significant toxicity despite profound neutropenia. We retrieved the previous reports of myelosuppressive therapies after IFD using various secondary prophylaxes, and showed that voriconazole is the most effective drug to suppress IFD relapses.

[Sensitization and crossreaction of simple coumarins].

The contact sensitization of 11 simple coumarins was examined by subcutaneous sensitizing of guinea pigs, and the structure-activity relationship and cross-reactivity were investigated. Esculetin, 4-methylesculetin, and dephnetin were found to be strong sensitizers, and 4-hydroxy-coumarin to be a moderate sensitizer. Other simple coumarins tested had a weak sensitivity to mild sensitizers. The results suggest that the introduction of hydroxy group, especially adjacent substitution at the 6, 7, and 8 positions of the coumarin ring with two hydroxy groups, may play an important role in exhibiting the contact sensitization activity. The cross-reactivity was observed between esculetin and 4-methylesculetin, esculin or isoscoporetin, and also between daphnetin and 4-methylumbelliferone or umbelliferone, although there was no mutual cross-reactivity between esculetin and daphnetin. It is interesting to note that guinea pigs, which had a weak sensitivity to umbelliferone, showed a strong cross-reactivity to daphnetin, while those, which had a weak sensitivity to daphnetin, showed a weak cross-reactivity to umbelliferone. It is assumed that a skin-protein conjugation at 5 or 6 positions of the coumarin ring is important to elicit the cross-reactivity of esculetin or daphnetin groups.

Vegetarian diet ameliorates symptoms of atopic dermatitis through reduction of the number of peripheral eosinophils and of PGE2 synthesis by monocytes.

Many patients with atopic dermatitis are dissatisfied with conventional treatments based on topical steroids and have experienced some traditional remedies and alternative therapies. However, most of such therapies have not been evaluated scientifically and clinically by specialists. This study was designed to assess whether a certain vegetarian diet might be effective for atopic dermatitis and if so, to identify the mechanisms of this remedy through analyses of immunological parameters. An open-trial study was carried out in twenty patients with atopic dermatitis. An improvement of dermatitis was evaluated by SCORAD index and serological and immunological parameters were monitored. After a two-month treatment, the severity of dermatitis was strikingly inhibited, as assessed by SCORAD index and serological parameters including LDH5 activity and a number of peripheral eosinophils. A sharp reduction in eosinophils and neutrophils was observed prior to improvement in the skin inflammation. In addition, PGE2 production by peripheral blood mononuclear cells was reduced by this treatment. In contrast, serum IgE levels did not change during the same period. Although this study is an open-trial one, it suggests that this treatment may be useful for the treatment of adult patients with severe atopic dermatitis.

8-hydroxydeoxyguanosine in urine as an index of oxidative damage to DNA in the evaluation of atopic dermatitis.

8-hydroxydeoxyguanosine (8-OHdG) is one of the products which are excreted in urine as a result of oxidative damage to DNA. We investigated the feasibility of using 8-OHdG in urine as an index for oxidative damage to DNA in atopic dermatitis (AD). Seventeen patients with long-standing AD and 17 healthy volunteers were enrolled in this study. The severity of AD was evaluated by SCORAD index. Eosinophils, total IgE and lactate dehydrogenase-5 in peripheral blood were measured as clinical parameters for AD. A newly developed enzyme-linked immunosorbent assay method was used to measure urine 8-OHdG. The AD patients showed significantly higher levels (P < 0.0001) of 8-OHdG in their urine than corresponding controls. Urine 8-OHdG levels showed as strong a positive correlation as other haematological parameters did using the SCORAD index. Thus, we conclude that the urine 8-OHdG levels can also serve as a biochemical index of tissue damage and can act as a useful tool in the clinical evaluation of AD.

Sequence analysis of the Porphyromonas gingivalis dipeptidyl peptidase IV gene.

We previously constructed a Porphyromonas gingivalis genomic library and isolated the 2.9 kb EcoRV fragment which specified glycylprolyl dipeptidyl aminopeptidase (GPase). Nucleotide sequencing of this fragment identified the single 2169 bp open reading frame which coded for a 723 amino acid protein. The amino acid sequencing of the NH2-terminal domain of the native and recombinant mature enzymes suggested that the protease possessed a 16 amino acid residue signal peptide. The calculated mass of the precursor and mature proteases were 82,018 and 80,235 daltons, respectively. The homology search of this enzyme in registered protein sequences revealed that this enzyme was homologous to dipeptidyl peptidase (DPP) IV from the Flavobacterium meningosepticum and that from eukaryotic cells, including the human, mouse, and rat. Three amino acid residues, Ser-593, Asp-668, and His-700, were identified as a putative catalytic triad, a common feature of eukaryotic serine proteases. In addition, this enzyme showed a broad proteolytic spectrum toward synthetic substrates capable of splitting not only Gly-Pro-derivative but also Ala-Pro, Lys-Pro, and Phe-Pro-derivatives. Therefore, we conclude that this enzyme belongs to DPP IV rather than GPase.

An alternative study of the skin irritant effect of an homologous series of surfactants.

An attempt has been made to differentiate between in vivo and in vitro skin reactions to a homologous series of surfactants (sodium alkyl sulfate, R-OSO(3) Na) and to determine the usefulness of percutaneous absorption in vitro as an alternative test system. Sodium alkyl sulfate showed considerable biological activity by virtue of its polar head groups. The length of the lipophilic chain in the surfactants was an important factor in their overall activity. The following in vivo tests were performed: a primary skin irritation test in guinea pigs, a primary eye irritation test in rabbits and a closed patch test in humans. Peak skin irritation occurred with C(10)-C(16) sodium alkyl sulfate, which had lipophilic groups of different alkyl chain lengths. Cell injury was also evaluated by the neutral red dye uptake assay in rabbit corneal (RC) cells. C(4) and C(6) compounds had no effect, while maximal effects occurred with C(18). Protein denaturation and haemolysis occurred with C(10)-C(16) compounds. In the percutaneous absorption test in guinea pig skin, permeation was low for the C(18) compound and high for the C(4) compound. The results with the C(18) compound suggest that differences between cell injury and skin irritation result from skin permeation. Although the C(18) compound caused cell injury, membrane destruction and protein denaturation were more severely with the C(10)-C(16) compounds, owing to their strong haemolytic and protein-denaturation action.

Cloning of the gene encoding a glycylprolyl aminopeptidase from Porphyromonas gingivalis.

A genomic library of Porphyromonas gingivalis 381 was constructed in the cosmid vector pHC79. A clone, pSN1, was identified by the expression of glycylprolyl-naphthylamide hydrolysing activity. The DNA insert contained within the cosmid pSN1 was subcloned into the plasmid vector pBR328 to create the recombinant plasmid pSN11 containing a 2.9 kb EcoRV insert. An Escherichia coli transformant containing pSN11 produced a protein having a molecular weight of 75 kDa. Southern-blot hybridization revealed that the 2.9 kb EcoRV DNA hybridized with an identical sized Eco RV DNA fragment in the chromosomal DNA of P. gingivalis 381.

[Methods for optimized cultivation of hair cells from C3H mice].

Methods for the optimized culturing the hair cells from 4-day-old C3H mice are described. The skin was obtained and soaked in 500 unit/ml dispase at 4 degrees C for 24 hr. Then the epidermis was removed and the dermis was further treated with 0.25% collagenase for 1 hr at 37 degrees C. The hair roots isolated by collagenase digestion of the dermis were dispersed by the treatment with the mixture of trypsin and EDTA into a cell suspension and plated on a substrate. Attachment of hair cells was dependent on serum concentrations and enhanced by collagen coating of the surface of dishes. MCDB 153 was beneficial for the growth of hair cells and discouraged for the growth of dermal fibroblasts. The cells could not grow at inoculum size of less than 3 x 10(4) cells/cm2. Addition of a crude bovine pituitary extract in MCDB 153 had the greatest impact on hair cell growth. Functional integrity of the cultured hair cells were maintained, since the hair-specific cytoskeletal proteins were expressed in these cells under the present experimental conditions. These desirable conditions could allow selective growth of the hair cells and provide the model system which could be used for the study of hair growth.

Micronucleus test with methyl methanesulfonate administered by intraperitoneal injection and oral gavage.

The effects of 2 routes of administration, intraperitoneal injection (i.p.) and oral gavage (p.o.), in the micronucleus test were evaluated using methyl methanesulfonate (MMS) and 2 strains of mice (MS/Ae and CD-1). A small-scale acute toxicity study and a pilot micronucleus experiment were carried out first. On the basis of the results obtained, a final micronucleus test was performed at doses of 20, 40, 80, and 160 mg/kg (i.p.) and 40, 80, 160, and 320 mg/kg (p.o.), with a 24-h sampling time. MMS induced micronucleated polychromatic erythrocytes (MNPCEs) in both routes in both mouse strains under the conditions used. At 40 and 80 mg/kg, MMS induced a higher number of MNPCEs by the i.p. route in both strains. A 160 mg/kg MMS dose induced higher numbers of MNPCEs by the p.o. route in MS/Ae mice. The route-related difference with MMS on the basis of mg/kg disappeared when the difference was determined on the basis of a ratio of the LD50. In practice, both i.p. and p.o. routes are acceptable as routes of administration in the micronucleus test using this chemical.