Evaluation of a Capillary Microsampling Device for Analyzing Plasma Lenvatinib Concentration in Patients With Hepatocellular Carcinoma.

BACKGROUND: The anticancer drug, Lenvima (lenvatinib), has severe side effects. Therapeutic drug monitoring helps ensure its efficacy and safety. Regular and optimally timed blood sampling is tough, especially when lenvatinib is self-medicated. Microsampling using the easy to handle Microsampling Wing (MSW) may help circumvent this problem. However, current lenvatinib detection methods are not sensitive enough to detect its concentrations in microsamples (<50-250 µL). Thus, the aim of this study was 2-fold (1) develop an analytic method to estimate plasma lenvatinib concentrations in microsamples and (2) verify whether this method works on micro (5.6 µL) blood plasma samples obtained clinically through MSW from patients with unresectable hepatocellular carcinoma (HCC). METHODS: A simple, highly sensitive, and specific liquid chromatography-electrospray ionization tandem mass spectrometry method was developed. Using this novel protocol, the trough blood plasma concentration of lenvatinib was measured for both blood sampled conventionally and that using MSW. Thirty-five venous whole blood samples were obtained from 11 patients with HCC. Furthermore, the stability of lenvatinib in MSW samples during storage was evaluated. RESULTS: The mean plasma lenvatinib concentration estimates were not significantly different between the MSW and conventional venous blood samples. CV for interday and intraday assays was low. Up to day 5, the lenvatinib concentration in the MSW samples was 85%-115% of the initial day concentration (when stored at 25°C or 4°C). The interference of endogenous matrix components in the human plasma was low. CONCLUSIONS: These results indicate that the novel mass spectrometry protocol accurately measures lenvatinib in human plasma and is reproducible. Thus, MSW could be a useful microsampling device for lenvatinib therapeutic drug monitoring in patients with HCC when used in combination with this novel liquid chromatography-electrospray ionization tandem mass spectrometry detection method.

Biomechanical Analysis of Unstable Osteochondral Fragment Fixation Using Three Different Techniques: Osteochondral Plug, Bioabsorbable Pin, and Suture Anchor with Tape.

Purpose: To compare the initial fixation strength of osteochondral fragment fixations using osteochondral plugs, bioabsorbable pins, and knotless suture anchors. Methods: Eighteen fresh-frozen immature (6 month old) porcine knees were used. An osteochondral fragment, cut from the articular surface of the medial femoral condyle to achieve a thickness of 5 mm, was used to mimic the unstable osteochondral fragment. It was fixed using three techniques, including two osteochondral plugs (osteochondral plug group), four full-threaded poly l-lactic acid pins (bioabsorbable pin group), and three suture anchors with a 2-0 tape (suture anchor group). Tensile loads at displacements of 1 and 2 mm and ultimate failure load were measured at a cross-head speed of 100 mm/min, and the variables of the three groups were compared statistically using a one-way ANOVA with Tukey's honestly significant difference test. Results: There was no significant difference in the tensile load to achieve 1-mm displacement. The load to achieve 2-mm displacement and the ultimate failure load were significantly greater in the suture anchor group than the osteochondral plug group and the bioabsorbable pin group. Conclusions: Single-pull destructive testing of a fixed articular osteochondral fragment with the force perpendicular to the articular surface, demonstrated no statistical difference in the tensile load to achieve 1-mm displacement, but the load to achieve 2-mm displacement was significantly greater for the three suture anchor-interlocking 2-0 tape constructs than the dual osteochondral plug fixation and the four bioabsorbable pin fixation constructs. Additionally, the three suture anchor-interlocking 2-0 tape construct's mean single-pull failure load was greater than other two fixation procedures. Clinical Relevance: To achieve osteochondral fragment union, sufficient fixation strength is critical. However, the initial fixation strength of osteochondral plugs, bioabsorbable pins, and knotless suture anchors for unstable osteochondral lesions remains unclear.

Internal Fixation of Osteochondritis Dissecans Using PushLock Suture Anchors.

Osteochondritis dissecans (OCD) of the knee is a subchondral bone abnormality that results in the separation of the articular cartilage and bone with subsequent progression to osteoarthritis. Unstable OCD lesions should undergo fixation to preserve the natural contour of the articular surface. Although several fixation procedures have been reported, the appropriate procedure remains unknown. Because the bony portion of the OCD lesion is usually thin, it is difficult to fix firmly with conventional methods. We began fixing OCD lesions with knotless PushLock anchors and sutures and have obtained satisfactory results. This report describes this fixation method that uses the PushLock suture anchor to treat unstable OCD lesions. This procedure also can be applied for traumatic osteochondral fractures.

Transcriptional Induction of Cystathionine γ-Lyase, a Reactive Sulfur-Producing Enzyme, by Copper Diethyldithiocarbamate in Cultured Vascular Endothelial Cells.

As toxic substances can enter the circulating blood and cross endothelial monolayers to reach parenchymal cells in organs, vascular endothelial cells are an important target compartment for such substances. Reactive sulfur species protect cells against oxidative stress and toxic substances, including heavy metals. Reactive sulfur species are produced by enzymes, such as cystathionine γ-lyase (CSE), cystathionine ß-synthase, 3-mercaptopyruvate sulfurtransferase, and cysteinyl-tRNA synthetase. However, little is known about the regulatory mechanisms underlying the expression of these enzymes in vascular endothelial cells. Bio-organometallics is a research field that analyzes biological systems using organic-inorganic hybrid molecules (organometallic compounds and metal coordinating compounds) as molecular probes. In the present study, we analyzed intracellular signaling pathways that mediate the expression of reactive sulfur species-producing enzymes in cultured bovine aortic endothelial cells, using copper diethyldithiocarbamate (Cu10). Cu10 selectively upregulated CSE gene expression in vascular endothelial cells independent of cell density. This transcriptional induction of endothelial CSE required both the diethyldithiocarbamate scaffold and the coordinated copper ion. Additionally, the present study revealed that ERK1/2, p38 MAPK, and hypoxia-inducible factor (HIF)-1α/HIF-1ß pathways mediate transcriptional induction of endothelial CSE by Cu10. The transcription factors NF-κB, Sp1, and ATF4 were suggested to act in constitutive CSE expression, although the possibility that they are involved in the CSE induction by Cu10 cannot be excluded. The present study used a copper complex as a molecular probe to reveal that the transcription of CSE is regulated by multiple pathways in vascular endothelial cells, including ERK1/2, p38 MAPK, and HIF-1α/HIF-1ß. Bio-organometallics appears to be an effective strategy for analyzing the functions of intracellular signaling pathways in vascular endothelial cells.

Permanent prostate brachytherapy with or without supplemental external beam radiotherapy as practiced in Japan: outcomes of 1300 patients.

PURPOSE: To report outcomes for men treated with iodine-125 ((125)I) prostate brachytherapy (BT) at a single institution in Japan. METHODS AND MATERIALS: Between 2003 and 2009, 1313 patients (median age, 68 years) with clinically localized prostate cancer were treated with (125)I BT. Median prostate-specific antigen level was 7.6 ng/mL (range, 1.1-43.3). T-stage was T1c in 60%, T2 in 39%, and T3 in 1% of patients. The Gleason score was <7, 7, and >7 in 49%, 45%, and 6% of patients, respectively. Neoadjuvant androgen deprivation therapy was used in 40% of patients and combined external beam radiotherapy of 45 Gy in 48% of patients. Postimplant dosimetry was performed after 30 days after implantation, with total doses converted to the biologically effective dose. Survival functions were calculated by the Kaplan-Meier method and Cox hazard model. RESULTS: Median followup was 67 months (range, 6-126). The 7-year biochemical freedom from failure for low-, intermediate-, and selected high-risk prostate cancers were 98%, 93%, and 81%, respectively (p < 0.001). Multivariate analysis identified the Gleason score, initial prostate-specific antigen level, positive biopsy rate, dose, and neoadjuvant androgen deprivation therapy as predictors for biochemical freedom from failure. The 7-year actuarial developing Grade 3+ genitourinary and gastrointestinal toxicity was 2% and 0.3%, respectively. Forty-four percent patients with normal baseline potency retained normal erectile function at 5 years. CONCLUSIONS: (125)I prostate BT is a highly effective treatment option for low-, intermediate-, and selected high-risk prostate cancers. Side effects were tolerable. An adequate dose may be required to achieve successful biochemical control.

Dietary intake in Japanese patients with type 2 diabetes: Analysis from Japan Diabetes Complications Study.

AIMS/INTRODUCTION: Though there are many differences in dietary habits and in the metabolic basis between Western and Asian people, the actual dietary intake in Asian patients with diabetes has not been investigated in a nationwide setting, unlike in Western countries. We aimed to clarify dietary intake among Japanese individuals with type 2 diabetes, and identify differences in dietary intake between Japanese and Western diabetic patients. MATERIALS AND METHODS: Nutritional and food intakes were surveyed and analyzed in 1,516 patients with type 2 diabetes aged 40-70 years from outpatient clinics in 59 university and general hospitals using the food frequency questionnaire based on food groups (FFQg). RESULTS: Mean energy intake for all participants was 1737 ± 412 kcal/day, and mean proportions of total protein, fat, and carbohydrate comprising total energy intake were 15.7, 27.6 and 53.6%, respectively. They consumed a 'low-fat energy-restricted diet' compared with Western diabetic patients, and the proportion of fat consumption was within the suggested range that has been traditionally recommended in Western countries. As a protein source, consumption of fish (100 g) and soybean products (71 g) was larger than that of meat (50 g) and eggs (29 g). These results imply that dietary content and food patterns among Japanese patients with type 2 diabetes are quite close to those reported as suitable for prevention of obesity, type 2 diabetes, cardiovascular disease, and total mortality in Europe and America. CONCLUSIONS: A large difference was shown between dietary intake by Japanese and Western patients. These differences are important to establish ethnic-specific medical nutrition therapy for diabetes.

Global brain delivery of neprilysin gene by intravascular administration of AAV vector in mice.

Accumulation of amyloid-ß peptide (Aß) in the brain is closely associated with cognitive decline in Alzheimer's disease (AD). Stereotaxic infusion of neprilysin-encoding viral vectors into the hippocampus has been shown to decrease Aß in AD-model mice, but more efficient and global delivery is necessary to treat the broadly distributed burden in AD. Here we developed an adeno-associated virus (AAV) vector capable of providing neuronal gene expression throughout the brains after peripheral administration. A single intracardiac administration of the vector carrying neprilysin gene in AD-model mice elevated neprilysin activity broadly in the brain, and reduced Aß oligomers, with concurrent alleviation of abnormal learning and memory function and improvement of amyloid burden. The exogenous neprilysin was localized mainly in endosomes, thereby effectively excluding Aß oligomers from the brain. AAV vector-mediated gene transfer may provide a therapeutic strategy for neurodegenerative diseases, where global transduction of a therapeutic gene into the brain is necessary.