Experimental and comparative studies suggest that the striped coats of zebras can prevent biting fly attacks. Biting flies are serious pests of livestock that cause economic losses in animal ...production. We hypothesized that cows painted with black and white stripes on their body could avoid biting fly attacks and show fewer fly-repelling behaviors. Six Japanese Black cows were assigned to treatments using a 3 × 3 Latin-square design. The treatments were black-and-white painted stripes, black painted stripes, and no stripes (all-black body surface). Recorded fly-repelling behaviors were head throw, ear beat, leg stamp, skin twitch, and tail flick. Photo images of the right side of each cow were taken using a commercial digital camera after every observation and biting flies on the body and each leg were counted from the photo images. Here we show that the numbers of biting flies on Japanese Black cows painted with black-and-white stripes were significantly lower than those on non-painted cows and cows painted only with black stripes. The frequencies of fly-repelling behaviors in cows painted with black-and-white stripes were also lower than those in the non-painted and black-striped cows. These results thus suggest that painting black-and-white stripes on livestock such as cattle can prevent biting fly attacks and provide an alternative method of defending livestock against biting flies without using pesticides in animal production, thereby proposing a solution for the problem of pesticide resistance in the environment.
Abstract Aims The aim of this study was to examine whether percutaneous electrical muscle stimulation (EMS) attenuates postprandial hyperglycemia in type 2 diabetes. Methods Eleven patients with type ...2 diabetes participated in two experimental sessions; one was a 30-min EMS 30 min after a breakfast (EMS trial) and the other was a complete rest after a breakfast (Control trial). In each trial, blood was sampled before and at 30, 60, 90, and 120 min after the meal. Results Postprandial glucose level was significantly attenuated in EMS trial at 60, 90, and 120 min after a meal ( p < 0.05). The C-peptide concentration was also significantly lowered in EMS trial ( p < 0.01). On the other hand, there was no significant increase in creatine phosphokinase (CPK) concentration in each trial. Conclusions The present results provide first evidence indicating that EMS is a new exercise method for treating postprandial hyperglycemia in individuals with type 2 diabetes, especially who cannot perform adequate voluntary exercise because of excessive obesity, orthopedic diseases, or severe diabetic complications.
We characterized leukemic cells from 20 adult T-cell leukemia (ATL) cases and 7 ATL-derived cell lines in terms of Foxp3 messenger RNA (mRNA) expression, cytokine production, cell surface markers ...associated with regulatory T-cells (Treg), and in vitro immunoregulatory activity and compared the results with those of cells from 3 T-cell-type chronic lymphocytic leukemia (T-CLL) patients and normal CD4+ T-cells. Real-time polymerase chain reaction analysis showed that cells from 10 ATL cases, 1 T-CLL case, and 1 ATL cell line had higher Foxp3 mRNA levels than CD4+ T-cells. In 5 ATL cases, Foxp3 levels were comparable to those of CD4+CD25+ T-cells. Flow cytometric analysis revealed that CTLA-4 expression correlated with Foxp3 mRNA level in ATL cells. The cells of all ATL cases examined produced no interleukin 2 or interferon gamma after iono-mycin and phorbolmyristate acetate stimulation. Cases with low Foxp3 expression (Foxp3-low) tended to express higher levels of transforming growth factor beta mRNA, but this trend was not statistically significant. An in vitro inhibition assay showed that the proliferation of normal CD4+CD25- T-cells stimulated with anti-CD3 monoclonal antibody and autologous dendritic cells was significantly suppressed by coculture with Foxp3-high ATL cells. These results indicate that Foxp3 expression is variable in ATL cases and that Foxp3-high ATL cells, which resemble Treg phenotypically as well as functionally, may be involved in immune suppression in ATL.
In this study, we attempted a comprehensive analysis of MHC class I–bound peptides in adult T cell leukemia (ATL) cells in order to identify as many tumor-associated antigens (TAAs) as possible that ...could be used for CTL-based immunotherapy.
Using mass spectrometry combined with reversed-phase liquid chromatography, we could sequence 188 HLA class I–restricted candidate peptides from three ATL-derived cell lines. In accordance with the restrained expression of HTLV-I viral RNA in these cell lines, there were no HTLV-I-encoded peptides among these candidates. Based on the differential expression between ATL cells and normal CD4
+ T cells, we selected 10 novel peptides as T cell epitopes of overexpressed source proteins. RT-PCR analysis revealed that 5 source proteins including PRAME, a known tumor-testis antigen, were highly expressed in the majority of 16 ATL cases. Furthermore we could induce PRAME-specific CTLs in vitro from an HLA-B62
+ healthy donor that showed specific cytotoxicity against HLA-B62
+ PRAME
+ ATL cells.
These results demonstrate that comprehensive analysis of HLA class I–bound peptides by mass spectrometry is useful for identification of TAA-derived peptides in ATL. Considering that expression patterns of leukemia/lymphoma-associated antigens vary from case to case, this approach appears to be suitable for the tailor-made immunotherapy of hematological malignancies.
In the present study, we attempted a comprehensive analysis of human leukocyte antigen (HLA) class I-bound peptides presented on adult T-cell leukemia (ATL) cells by the latest technology of mass ...spectrometry combined with reversed phase liquid chromatography (LC/MS) to identify novel tumor-associated antigens. We screened the sequenced peptides for those compatible with the motives of the respective HLA class I alleles. Then, we narrowed down the candidate peptides according to the differential expression of their source proteins between ATL cells and normal CD4 T cells. Among these candidates, we focused on cyclin-dependent kinaselike 5 (CDKL5) because it was highly expressed in several ATL cell lines and some ATL clinical samples but not in normal CD4 T cells. To examine its immunogenicity, we stimulated CD8 T cells from an HLA-B62 healthy donor several times with autologous monocyte-derived dendritic cells loaded with HLA-B*62-restricted CDKL5 peptide1012-1021 QVNQAALLTY that we identified. CDKL5-stimulated bulk CD8 T cells exerted higher cytotoxicity against CDKL5 peptide-loaded autologous Epstein Barr virus-transformed B cell line (LCL) than against unloaded LCL. Furthermore these T cells had strong cytotoxic activity against HLA-B*62-positive CDKL5-positive but not HLA-B*62-negative CDKL5-positive ATL cells. These results demonstrate that CDKL5 is a novel tumor (leukemia) antigen in ATL and that the HLA-B*62-restricted CDKL5 peptide can be used for cytotoxic T-lymphocyte-mediated immunotherapy. Identification of tumor-associated antigens by LC/MS is an eligible and efficient method suitable for future taylor-made immunotherapy of hematologic malignancies.
Summary
Recent studies have shown that human myeloid leukaemia cells can differentiate into dendritic cell (DC)‐like cells (leukaemia‐DCs) when cultured with a combination of cytokines. In the ...present study, we examined whether the transduction of leukaemia‐DCs with OX40 ligand (OX40L), a member of the tumour necrosis factor (TNF) family, resulted in augmentation of their antigen presenting activity. Bicistronic retroviral vectors expressing both human OX40L and enhanced green fluorescent protein (EGFP) or EGFP alone were generated and used for transduction. Fresh leukaemic cells from five patients with acute myeloid leukaemia (AML) were isolated and retrovirally transduced with OX40L during the culture with a combination of cytokines from stem cell factor, fms‐like tyrosine kinase (Flt)‐3 ligand, granulocyte‐macrophage colony stimulating factor (GM‐CSF), interleukin‐4 (IL‐4) and TNF‐α. After 7 d, the majority of cells showed DC‐like morphology, and expressed higher levels of CD80, CD86 and HLA‐DR than fresh leukaemic cells. The transduction efficiency was 8·5–27·2%. Leukaemia‐DCs transduced with OX40L elicited higher proliferative response of allogeneic CD4+ T cells than fresh leukaemic cells, non‐transduced, or mock‐transduced leukaemia‐DCs. Co‐culture of allogeneic CD4+ T cells with OX40L‐transduced leukaemia‐DCs was superior in the generation of interferon (IFN)‐γ producing CD4+ T cells and in production of IFN‐γ. Furthermore, OX40L‐transduced leukaemia‐DCs could elicit significant proliferative response of human leucocyte antigen‐matched T cells from the donor in allogeneic stem cell transplantation. These results indicate that retroviral transduction of leukaemia‐DCs with OX40L augments their antigen presenting cell activity and thus renders them more suitable for tumour vaccines or ex vivo stimulation of leukaemia‐specific T cells.
Electron beam lithography (EBL) is very important for semiconductor manufacturing process, especially for making photo-masks and ASIC devices. EBL is also important for fabricating MEMS, MOES and so ...on. In this paper, we used hydrogen silsesquioxane (HSQ), which acts as a negative resist (Namatsu, 1998). And we fabricated three-dimensional HSQ resist structure by changing the EB acceleration voltage.
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