Search

Microscale needle technology is important in electrophysiological studies, drug/chemical delivery systems, optogenetic applications, and so on. In this study, dissolvable needle-base scaffold realizes penetration of high-aspect-ratio flexible microneedles (e.g., <5 μm diameter and >500 μm length) into biological tissues. This methodology, which is applicable to numerous high-aspect-ratio flexible microneedles, should reduce the invasiveness and provide safer tissue penetrations than conventional approaches., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Phosphoglycerate kinase (PGK) deficiency, a rare X-linked inherited disorder, manifests as various combinations of hemolytic anemia, neurological dysfunction, and myopathy. We report a Japanese boy with PGK deficiency presenting as chronic hemolytic anemia. The diagnosis of PGK1 deficiency was made at 11 months of age on the basis of low PGK enzyme activity (36.7 IU/g Hb; normal, 264-326 IU/g Hb) and the identification through PGK1 gene sequencing of a novel missense mutation: c. 1180A>G at exon 10. The mutation, which has been designated PGK-Aoto, results in a Thr394Ala amino-acid substitution at β-strand L. Because β-strand L plays an important role in the function of the hinge connecting the two domains of PGK, the Thr394Ala substitution may perturb this motion. At 3 years of age the patient has transfusion-dependent hemolytic anemia but no evidence of neuromuscular disease or developmental delay. Long-term follow-up will be needed to identify possible future clinical manifestations.

Esophagus squamous cell carcinoma (ESCC) is one of the most deadly malignances because of its high frequency of metastasis. Given the associations of MUC1 with ESCC and tumor metastasis, we explored a potential role of MUC1 in ESCC metastasis. Among 40 ESCC and 20 paired normal tissue specimens examined, we found a significant increase of MUC1 expression in ESCC and more importantly, that expression of MUC1 and MMP13 are strongly correlated in patients who had lymph node metastasis. Studies with cell models indicated that overexpression of MUC1 upregulates the expression of MMP13, leading to increased cell migration. In support of a mode of transcriptional regulation, promoter analysis revealed that MUC1 stimulates MMP13 expression through the Runx-2-binding site. The link of MUC1 to cell motility was further confirmed by the finding that depletion of MUC1 resulted in reduced expression of MMP13 and cell migration, invasion and adhesion. Moreover, the loss of cell metastatic potential was rescued by overexpression of MMP13 completely. Collectively, our findings indicate that MUC1 contributes to ESCC metastasis by stimulating MMP13 expression, suggesting MUC1 as a novel diagnostic biomarker and therapeutic target in ESCC.

Although p53 is intact in most cases of retinoblastoma, it is largely inactivated by the ubiqutin-proteasome system through interaction with murine double minute 2 (MDM2) and murine double minute X (MDMX). The present study showed that the histone deacetylase (HDAC) inhibitors valproic acid (VPA) and depsipeptide (FK228) synergistically enhanced ionizing radiation (IR)-induced apoptosis, associated with activation of caspase-3 and cleavage of poly(ADP-ribose) polymerase in Y79 and WER1-Rb1 human retinoblastoma cells. Both VPA and FK228 enhanced IR-induced phosphorylation of histone H2AX on Ser139 preceding apoptosis. Exposure of cells to IR in the presence of VPA or FK228 induced the accumulation of p53 acetylated at Lys382 and phosphorylated at Ser46 through the reduction of binding affinity with MDM2 and MDMX. These results suggest that acetylation of p53 by HDAC inhibitors is a promising new therapeutic target in refractory retinoblastoma.

Nuclear factor-kappaB (NF-kappaB) is constitutively activated in diverse human malignancies. The mucin 1 (MUC1) oncoprotein is overexpressed in human carcinomas and, like NF-kappaB, blocks cell death and induces transformation. The present studies show that MUC1 constitutively associates with NF-kappaB p65 in carcinoma cells. The MUC1 COOH-terminal subunit (MUC1-C) cytoplasmic domain binds directly to NF-kappaB p65 and, importantly, blocks the interaction between NF-kappaB p65 and its inhibitor IkappaBalpha. We show that NF-kappaB p65 and MUC1-C constitutively occupy the promoter of the Bcl-xL gene in carcinoma cells and that MUC1-C contributes to NF-kappaB-mediated transcriptional activation. Studies in nonmalignant epithelial cells show that MUC1-C interacts with NF-kappaB in the response to tumor necrosis factor-alpha stimulation. Moreover, tumor necrosis factor-alpha induces the recruitment of NF-kappaB p65-MUC1-C complexes to NF-kappaB target genes, including the promoter of the MUC1 gene itself. We also show that an inhibitor of MUC1-C oligomerization blocks the interaction with NF-kappaB p65 in vitro and in cells. The MUC1-C inhibitor decreases MUC1-C and NF-kappaB p65 promoter occupancy and expression of NF-kappaB target genes. These findings indicate that MUC1-C is a direct activator of NF-kappaB p65 and that an inhibitor of MUC1 function is effective in blocking activation of the NF-kappaB pathway.

Background: Interferon (IFN) potentiates cytotoxicity by X-ray irradiation. To elucidate the mechanism of this potentiation, the biological markers related to DNA damage and cell survival were studied., Materials and Methods: IFN-alpha-sensitive Daudi and its resistant cells were used. Survival after treatment was assessed by clonogenic assays. DNA breaks were studied by pulse-field gel electrophoresis (PFGE). Production of reactive oxygen metabolites was measured using flow cytometry. Messenger RNA and protein were examined by RT-PCR and immunoblot, respectively., Results: IFN-alpha treatment for 24 h before irradiation potentiated the sensitivity of Daudi cells to X-rays. This combination induced 50 kb DNA fragmentation and activated caspase-3 in Daudi cells. Pretreatment with IFN-alpha inhibited the production of reactive oxygen species by irradiation. IFN-alpha pretreatment down-regulated most of the double-strand break (DSB) repair-related mRNAs, but did not affect the repair of DSBs studied by PFGE. The induction and phosphorylation of p21(Cip1/WAF1) (p21) was prominently suppressed in cells pretreated with IFN-a., Conclusion: Pretreatment with IFN-alpha potentiates the cytotoxic effects of X-rays. Inhibition of X-ray-induced p21 may cause the augmented sensitivity by IFN-alpha pretreatment.

Background: Imatinib mesylate (ST1571) is the first-line drugfor chronic myeloid leukemia (CML), but development of resistance to this drug is a clinical problem. To explore the effective use of ST1571, we studied the combination treatment with histone deacetylase inhibitor (depsipeptide, FK228)., Materials and Methods: FK228 and trichostatin A (TSA) were studied with respect to apoptosis of two Bcr-Abl-positive cell lines, K562 and TCC-S. Genetically-modified K562 cells by any of cyclin D1, c-Myc and active MEK genes were also studied. Apoptosis was examined by nuclear-morphology under a fluorescent microscope and by the expression of annexin V Changes of apoptosis-regulating genes and acetylated histone H4 were studied by immunoblot., Results: FK228 showed cytotoxicity at the nano-molar level. Combination treatment with STI571 and FK228 enhanced the induction of apoptosis significantly compared with each single treatment, although the histone acetylation level was not changed by the co-treatment. The combination treatment activated caspase-3 and cleaved PARP, but it did not induce any notable change in the expression of Bcl-XL, Bcl-2 and Bax compared with each single treatment. Enhanced apoptosis by the co-treatment was abrogated by ectopic expression of cyclin D1, c-Myc or active MEK CONCLUSION: The combination of FK228 with STI571 is a promising treatment for Bcr-Abl-positive CML, but the activation of the MEK/ERK pathway and its downstream target genes may bring resistance to the co-treatment in leukemic cells.

This paper reports on the development of neural recording chip device with penetrating Si microprobe electrode array using IC-process. The Si microprobe electrode array each with a few microns in diameter was grown at predetermined positions with interconnection-wirings. Controlling the diameter and the length of Si probes can be realized by a selective vapor-liquid-solid (VLS) growth. In this work, Si probes with 2 mum in diameter and 60 microm in length were fabricated, which were conductive-Si probes and they were encapsulated with SiO2 layers. To reduce the impedance of Si probes, the tips of Si probes were coated with a metal Au layer. As a result, penetrating Si microprobes measured in saline solution, showed impedance of the order of 300 k to 500 komega at 1 kHz. Packaging techniques for the probe chip were performed with a fluid-tight chamber and a flexible-printed-circuit of polyimide for neural recording experiments.

B-cell diseases are classified on the basis of the normal differentiation stages. We report here a case of a patient with a long history of leukocytosis, splenomegaly without lymphadenopathy, and hyperviscosity symptoms. Clinically, the patient's diagnosis was leukemic Waldenstrom macroglobulinemia. Chromosomal analysis revealed translocation t(2;7)(p11;q22) along with disease progression. Death occurred from pulmonary infection at 46 months after the initial presentation. At autopsy, malignant lymphocytes were found in the marginal areas of the spleen with spreading to the bone marrow and the liver. The histologic findings were consistent with splenic marginal zone lymphoma. We examined the sequences of the immunoglobulin V(H) gene in cells from the initial peripheral blood and from the spleen at autopsy and found that the sequences were identical and had no somatic hypermutation. Macroglobulinemia can occur in various B-cell disorders, including splenic marginal zone lymphoma, even with the transformation of unmutated B-lymphocytes.

Background: BCL10, a gene involved in the chromosomal translocation t(1;14)(p22;q32) found in mucosa-associated lymphoid tissue lymphoma (MALT lymphoma), has shown mutation in not only MALT lymphomas but also other lymphold tumors. However, the mutation rate remains controversial. One possible reason is variation in the source material (DNA or RNA), with most studies having been done using tumor DNA. Accordingly, we studied BCL10 mutations using tumor RNA., Materials and Methods: Fifty lymphoid malignancies (26 malignant lymphomas, 10 chronic lymphocytic leukemias and 14 acute lymphoblastic leukemias) were examined. Total RNA was extracted from the tumor cells and first-strand cDNA was subjected to single-strand conformation polymorphism and fragment length analysis. Then the results were confirmed by direct sequencing., Results: No mutations of the BCL10 gene were found in the 50 samples. There were four polymorphisms (3G to T, 24G to C, 485C to T and 638G to A). All samples showed at least one 24C allele., Conclusion: The BCL10 mutations studied are rare events at the RNA level and may not be associated with the mechanism of tumorigenesis in most lymphoid tumors.