Increased c­SRC expression is involved in acquired resistance to lenvatinib in hepatocellular carcinoma.

Lenvatinib, a multi-kinase inhibitor, serves a crucial role in the treatment of unresectable hepatocellular carcinoma (HCC). However, >50% of patients receiving lenvatinib therapy experience tumor growth or metastasis within 1 year, highlighting the need to address acquired resistance as a critical clinical challenge. To elucidate the factors associated with acquired resistance to lenvatinib, a lenvatinib-resistant HCC cell line (JHH-7_LR) was established by exposing a lenvatinib-sensitive HCC cell line, JHH-7, to lenvatinib. The changes in protein expression associated with the development of resistance were analyzed using a proteomic approach, detecting 1,321 proteins and significant changes in the expression of 267 proteins. Using Ingenuity Pathway Analysis bioinformatics software, it was revealed that the activity of multiple signaling pathways varied alongside the changes in expression of these proteins, and c-SRC was identified as a protein involved in a number of these signaling pathways, with its activity varying markedly upon the acquisition of resistance. When co-administering dasatinib, a c-SRC inhibitor, the partial restoration of lenvatinib sensitivity in the JHH-7_LR cell line was observed. The present study demonstrated that increased c-SRC expression was partially associated with HCC resistance to lenvatinib, suggesting that c-SRC inhibition could reduce the resistance of HCC to lenvatinib.

Development and application of a rapid and sensitive genotyping method for pharmacogene variants using the single-stranded tag hybridization chromatographic printed-array strip (STH-PAS).

Genetic polymorphisms contribute to inter-individual variability in the metabolism of multiple clinical drugs, including warfarin, thiopurines, primaquine, and aminoglycosides. A rapid and sensitive clinical assessment of various genome biomarkers is, therefore, required to predict the individual responsiveness of each patient to these drugs. In this study, we developed a novel genotyping method for the detection of nine pharmacogene variants that are important in the prediction of drug efficiency and toxicity. This genotyping method uses competitive allele-specific PCR and a single-stranded tag hybridization chromatographic printed-array strip (STH-PAS) that can unambiguously determine the presence or absence of the gene variant by displaying visible blue lines on the chromatographic printed-array strip. Notably, the results of our STH-PAS method were in 100% agreement with those obtained using standard Sanger sequencing and KASP assay genotyping methods for CYP4F2 gene deletion. Moreover, the results were obtained within 90 min, including the PCR amplification and signal detection processes. The sensitive and rapid nature of this novel method make it ideal for clinical genetic testing to predict drug efficacy and toxicity, and in doing so will aid in the development of individualized medicine and better patient care.

Modified Surgical Stent for Accurate TAD Placement.

The development of temporary anchorage devices (TADs) has allowed for the creation of anchorage in situations where there once was none. Many studies have suggested that the most significant cause of miniscrew failure is insufficient distance between the surface of a root and the miniscrew. In order to reduce the miniscrew failure rate, a modified surgical stent has been shown to not only increase TAD insertion accuracy but also to increase TAD success rates.

Functional characterization of 10 CYP4A11 allelic variants to evaluate the effect of genotype on arachidonic acid ω-hydroxylation.

Genetic variations in cytochrome P450 4A11 (CYP4A11) contributes to inter-individual variability in the metabolism of fatty acids such as arachidonic acid. CYP4A11 metabolizes arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE), which is important for the regulation of blood pressure. Polymorphisms in CYP4A11 are associated with susceptibility to hypertension. In this study, we evaluated the in vitro ω-hydroxylation of arachidonic acid by 10 CYP4A11 allelic variants, which cause amino acid substitutions in the encoded proteins. CYP4A11 variants were heterologously expressed in COS-7 cells and the kinetic parameters of arachidonic acid ω-hydroxylation were estimated. Among 10 CYP4A11 variants, 5 (CYP4A11-v1, CYP4A11-v2, CYP4A11-v3, CYP4A11-v4, and CYP4A11-v7) showed no or markedly lower activity compared to wild-type CYP4A11. This functional analysis of CYP4A11 variants could provide useful information for the effective prevention and treatment of hypertension.

Functional characterization of wild-type and 49 CYP2D6 allelic variants for N-desmethyltamoxifen 4-hydroxylation activity.

Genetic variations in cytochrome P450 2D6 (CYP2D6) contribute to interindividual variability in the metabolism of clinically used drugs, e.g., tamoxifen. CYP2D6 is genetically polymorphic and is associated with large interindividual variations in therapeutic efficacy and drug toxicity. In this study, we performed an in vitro analysis of 50 allelic variants of CYP2D6 proteins. Wild-type CYP2D6.1 and 49 variants were transiently expressed in COS-7 cells, and the enzymatic activities of the CYP2D6 variants were characterized using N-desmethyltamoxifen as a substrate. The kinetic parameters K(m), V(max), and intrinsic clearance (V(max)/K(m)) of N-desmethyltamoxifen 4-hydroxylation were determined. Among the 50 CYP2D6 variants, the kinetic parameters for N-desmethyltamoxifen 4-hydroxylation were determined for 20 CYP2D6 variants. On the other hand, the kinetic parameters of 30 CYP2D6 variants could not be determined because the amount of metabolite produced was at or below the detection limit at the lower substrate concentrations. Among them, 8 variants, i.e., CYP2D6.2, .9, .26, .28, .32, .43, .45, and .70, showed decreased intrinsic clearance at <50% of CYP2D6.1. The comprehensive in vitro assessment of CYP2D6 variants provides novel insights into allele-specific activity towards tamoxifen and may be valuable when interpreting in vivo studies.

[Current status of the criteria for discharging patients with pulmonary tuberculosis--results of questionnaire survey on the criteria for discharging patients from tuberculosis wards of the hospitals in Kanto area].

INTRODUCTION: Emphasis in treating patients with infectious pulmonary tuberculosis has come to be laid on the execution of reliable standard chemotherapy. As a result, hospitalization for a prolonged period has become unnecessary any more. However, few attempts have been made so far on the determination of discharging criteria. METHODS: We performed a questionnaire survey to hospitals with wards for tuberculosis in Kanto area and asked questions on the current status of discharging criteria. RESULTS: The effective response rate to the survey was 63.0 %. Sputum smear examination carried out mainly by Ziehl-Neelsen method and fluorescence method in 17.2% and 72.4 % of the hospitals, respectively. Sputum culture examination was carried out using mainly Ogawa medium and a liquid medium in 62.1% and 27.6% of the hospitals, respectively. Discharging criteria were standardized in 79.3% of hospitals. Negative sputum smear was used as the criterion for determining discharge in 11 sets of criteria. Negative sputum culture was used as the criterion for determining discharge in 17 sets of criteria. In the remaining one hospital, patients were to be discharged after 2-month treatment. There was no consistency in the procedure and the frequency of sputum examinations, how many negative results are needed to confirm negative status and the criteria for judgment. CONCLUSION: These results suggest that further evaluation must be made on the treatment outcome at each hospital, and the standard discharging criteria should be worked out taking into account the capacity of each hospital and the care situation of local community.

Comparative study of craniofacial morphology and bite force in Fijians and Japanese.

An anthropological survey was conducted in Fiji in 1994 and 1995 to study dental arch form, craniofacial morphology, and bite force of Fijians. Measurements were obtained from dental casts, cephalograms, and thin pressure-sensitive sheets (Dental Prescale®) for bite force analysis. Results were compared with those of Japanese. In every direction, the size of the dental arch in Fijians was larger than in Japanese. Fijians displayed longer palates, longer mandibles, and bimaxillary protrusion. There was no significant difference in upper and lower facial heights. FH to lower incisor angle in Fijians was significantly larger than in Japanese. Fijians were characterized by a small palatal plane angle, occlusal plane angle and mandibular plane angle, and were thus brachyfacial. The Japanese tended to be more dolichofacial. The distances from the Cd line to the pterygoid muscles, masseter muscles, and teeth in Fijians were significantly longer than in Japanese. Occlusal contact areas of Fijians were also greater than those of Japanese. The results indicate that the masticatory muscles and craniofacial morphologies supporting them would be better integrated in Fijians than in Japanese. Am. J. Hum. Biol. 10:63-72, 1998. © 1998 Wiley-Liss, Inc.