Formation of pseudo[3]rotaxanes containing calix-bis-crowns and secondary ammonium ions and their thermodynamic stabilities in a solution: preorganization by second macrocycle and nonallosteric behavior exhibited by large crown cavities

This paper describes the formation of pseudo[3]rotaxanes containing calix[4]-bis-crowns, exhibiting a 1,3-alternate conformation and large crown cavities, and secondary ammonium ions. The first and second association constants of pseudo[3]rotaxane formation are moderate (K1 = 175, K2 = 100 M−1) and are higher than that of the corresponding pseudo[2]rotaxane (K = 24 M−1), consisting of a calix[4]-mono-crown and the same secondary ammonium ion

Development of a near-infrared spectroscopic system for monitoring urine glucose level for the use of long-term home healthcare

We have been developing a new technique for measuring urine glucose concentration using near infrared spectroscopy (NIRS) in conjunction with the Partial Least Square (PLS) method. In the previous study, we reported some results of preliminary experiments for assessing feasibility of this method using a FT-IR spectrometer. In this study, considering practicability of the system, a flow-through cell with the optical path length of 10 mm was newly introduced. Accuracy of the system was verified by the preliminary experiments using urine samples. From the results obtained, it was clearly demonstrated that the present method had a capability of predicting individual urine glucose level with reasonable accuracy (the minimum value of standard error of prediction: SEP = 22.3 mg/dl) and appeared to be a useful means for long-term home health care. However, mean value of SEP obtained by the urine samples from ten subjects was not satisfactorily low (53.7 mg/dl). For improving the accuracy, (1) mechanical stability of the optical system should be improved, (2) the method for normalizing the spectrum should be reconsidered, and (3) the number of subject should be increased

Ferrocenylnaphthalene Diimide-Based Electrochemical Detection of Aberrant Methylation in hTERT Gene

Since aberrant methylation at CpG sites is linked to the silencing of tumor suppressor genes, DNA methylation analysis is important for cancer diagnosis. We developed ferrocenylnaphthalene diimide (FND), which has two ferrocenyl moieties at the substituent termini, as an electrochemical indicator for hybridized DNA duplexes. In this study, we attempted to detect aberrant methylation of human telomerase reverse transcriptase gene (hTERT), an efficient cancer marker, using FND-based hybridization coupled with electrochemical detection via a multi-electrode chip

Electrochemical telomerase assay for screening for oral cancer

Telomerase has long been known to be a marker for cancer. We have developed a new method of detecting it: the electrochemical telomerase assay (ECTA). We have previously confirmed that the assay is easier to do and more precise than the conventional telomeric repeat amplification protocol, which is currently the most widely used. Here we describe a pilot study made to establish a screening system for oral cancer using ECTA. We evaluated three types of clinical samples obtained from 44 patients with oral cancer and 26 healthy volunteers: exfoliated cells from the whole oral cavity, exfoliated cells from local lesions, and tissue from the lesion itself. The current increase ratio (Δi) obtained by ECTA was significantly higher in the oral cancer group for each type of sampling used. The threshold value for Δi was 19% when calculated by analysis of receiver-operating characteristic curves. Sensitivity and specificity values were 86% and 85% for cells from the oral cavity, 82% and 85% in cells from local lesions, and 95% and 92% in cells from the tumour itself, respectively. There were also no significant differences in sensitivity and specificity associated with age, size of tumour, site of lesion, or degree of malignancy. ECTA therefore seems to be a promising assay for screening for oral cancer

Screening for Oral Cancer Using Electrochemical Telomerase Assay

Electrochemical telomerase assay (ECTA) developed by our group was evaluated in an oral cancer screening using exfoliated oral cells and tissues obtained from patients of oral cancer, mucosa associated disease, or healthy volunteers. Telomerase activity from ECTA is correlated with hTERT mRNA expression level using a real‐time PCR and was increasing in the following order: healthy volunteer group<mucosa associated disease group<oral cancer group. Sensitivity and specificity of ECTA were 88% and 72%, respectively when used 17% of the threshold value based on the receiver operating characteristic curve in ECTA data

Oral Cancer Screening Based on Methylation Frequency Detection in hTERT Gene Using Electrochemical Hybridization Assay via a Multi‐electrode Chip Coupled with Ferrocenylnaphthalene Diimide

Ferrocenylnaphthalene diimide‐based electrochemical hybridization assay via a multi‐electrode chip was applied to detect the methylation frequency in the promoter region of human telomerase reverse transcriptase (hTERT) gene for clinical samples from tissues, local exfoliated oral cells from a lesion, or from entire oral cavity after their methylation specific PCRs. These methylation frequencies were increased with cancer progress as the following order: healthy volunteers, oral leukoplakia as precancerous lesion, and oral squamous cell carcinoma (OSCC). Operating characteristic analysis of the obtained current data doesn\u27t only give excellent discrimination ability of OSCC, but also of oral leukoplakia from healthy volunteers for all samples. Sensitivity and specificity was 95% and 90%, respectively, which is a comparable with methods in practical use

BCL6 inhibition ameliorates resistance to ruxolitinib in <i>CRLF2</i>-rearranged acute lymphoblastic leukemia

Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) is an intractable disease and most cases harbor genetic alterations that activate JAK or ABL signaling. The commonest subtype of Ph-like ALL exhibits a CRLF2 gene rearrangement that brings about JAK1/2-STAT5 pathway activation. However, JAK1/2 inhibition alone is insufficient as a treatment, so combinatorial therapies targeting multiple signals are needed. To better understand the mechanisms underlying the insufficient efficacy of JAK inhibition, we explored gene expression changes upon treatment with a JAK1/2 inhibitor (ruxolitinib) and found that elevated BCL6 expression was one such mechanism. Upregulated BCL6 suppressed the expression of TP53 along with its downstream cell cycle inhibitor p21 (CDKN2A) and pro-apoptotic molecules, such as FAS, TNFRSF10B, BID, BAX, BAK, PUMA, and NOXA, conferring cells some degree of resistance to therapy. BCL6 inhibition (with FX1) alone was able to upregulate TP53 and restore the TP53 expression that ruxolitinib had diminished. In addition, ruxolitinib and FX1 concertedly downregulated MYC. As a result, FX1 treatment alone had growth-inhibitory and apoptosis- sensitizing effects, but the combination of ruxolitinib and FX1 more potently inhibited leukemia cell growth, enhanced apoptosis sensitivity, and prolonged the survival of xenografted mice. These findings provide one mechanism for the insufficiency of JAK inhibition for the treatment of CRLF2-rearranged ALL and indicate BCL6 inhibition as a potentially helpful adjunctive therapy combined with JAK inhibition