High-risk pathogenic germline variants in blood relatives of BRCA1/2 negative probands.

BACKGROUND: Tailored, preventive cancer care requires the identification of pathogenic germline variants (PGVs) among potentially at-risk blood relatives (BRs). Cascade testing is carried out for BRs of probands who are positive for PGVs of an inherited cancer but not for negative probands. This study was conducted to examine the prevalence of PGVs for BRs of PGV-negative probands. METHODS: PGV prevalence was assessed for 682 BRs of 281 probands with BRCA1/BRCA2 wild-type hereditary breast and ovarian cancer (HBOC) syndrome. RESULTS: PGVs were discovered in 22 (45.8%) of the 48 BRs of the PGV-positive probands and in 14 (2.2%) of 634 BRs of the PGV-negative probands. Eleven PGVs on high-risk BRCA1, BRCA2, and TP53 genes were present only in BRs and not in the probands (probands vs BRs in Fisher exact test; p = 0.0104; odds ratio [OR] = 0.000 [0.000-0.5489 of 95% confidence interval]), partly due to the nature of the selection criteria. The enrichment of high-risk PGVs among BRs was also significant as compared with a non-cancer East Asian population (p = 0.0016; OR = 3.0791 [1.5521-5.6694]). PGV prevalence, risk class of gene, and genotype concordance were unaffected by the cancer history among BRs. CONCLUSION: These findings imply the necessity to construct a novel testing scheme to complement cascade testing.

Clinicopathological and imaging features of ductal carcinoma in situ in BRCA1/2 mutation carriers.

BACKGROUND: BRCA1/2-associated invasive breast cancer has been extensively studied. However, there are few reports of ductal carcinoma in situ (DCIS). OBJECTIVE: This study aimed to investigate the clinicopathological and imaging findings of DCIS in patients with BRCA1/2 mutations. METHODS: This was a single-institution, retrospective study. We identified patients diagnosed with DCIS with BRCA mutations between September 2003 and December 2020. Clinicopathological data and mammography (MG), magnetic resonance imaging (MRI), and ultrasound (US) findings were reviewed. RESULTS: We identified 30 cancers in 28 patients; 7 (25.0%) patients had BRCA1 mutations, and 21 (75.0%) had BRCA2 mutations. The median patient age was 42 years. Screening was the most common reason for the detection of DCIS (50.0%), followed by occult cancer diagnosed by pathological examination after risk-reducing mastectomy (26.7%). The nuclear grade was most often 1 (46.7%), and 93.3% were estrogen and/or progesterone receptor positive. The detection rates of MG, MRI, and US were 64.3%, 72.0%, and 64.0%, respectively. The most common imaging findings were calcification (100%) on MG, non-mass enhancement (88.9%) on MRI, and hypoechoic area (75.0%) on US. CONCLUSION: BRCA-associated DCIS was more strongly associated with BRCA2, and imaging features were similar to those of sporadic DCIS. Our results are helpful in informing surveillance strategies based on genotypes in women with BRCA mutations.

Prevalence of disease-causing genes in Japanese patients with BRCA1/2-wildtype hereditary breast and ovarian cancer syndrome.

Panel sequencing of susceptibility genes for hereditary breast and ovarian cancer (HBOC) syndrome has uncovered numerous germline variants; however, their pathogenic relevance and ethnic diversity remain unclear. Here, we examined the prevalence of germline variants among 568 Japanese patients with BRCA1/2-wildtype HBOC syndrome and a strong family history. Pathogenic or likely pathogenic variants were identified on 12 causal genes for 37 cases (6.5%), with recurrence for 4 SNVs/indels and 1 CNV. Comparisons with non-cancer east-Asian populations and European familial breast cancer cohorts revealed significant enrichment of PALB2, BARD1, and BLM mutations. Younger onset was associated with but not predictive of these mutations. Significant somatic loss-of-function alterations were confirmed on the wildtype alleles of genes with germline mutations, including PALB2 additional somatic truncations. This study highlights Japanese-associated germline mutations among patients with BRCA1/2 wildtype HBOC syndrome and a strong family history, and provides evidence for the medical care of this high-risk population.

Inhibitory Effects of Prolonged Vibratory Stimulus on the Maximal Voluntary Contraction Force and Muscle Activity of the Triceps Brachii: An Experimental Study.

OBJECTIVE: The purpose of this study was to quantify the effects of prolonged vibratory stimulus on the maximal voluntary contraction (MVC) force and muscle activity of the triceps brachii and to clarify the effective stimulus time. METHODS: Twenty-five healthy volunteers with a mean age of 21.4 years participated. A vibratory stimulus at 86 Hz was applied to the triceps brachii tendon for 5 and 10 minutes. Before and after these stimuli, the elbow extension MVC force was measured using a handheld dynamometer. Muscle activities of the lateral, long, and medial heads of the triceps brachii were also recorded by surface electromyography. RESULTS: The median MVC force significantly decreased to 82.7% after 5 minutes of vibratory stimulus and to 83.3% after 10 minutes of vibratory stimulus (P < .001). The median percentage of integrated electromyography of the triceps also significantly decreased to 78.2 (lateral head), 83.8 (long head), and 81.5 (medial head) after 5 minutes of vibratory stimulus and to 77.7, 81.4, and 77.2, respectively, after 10 minutes of vibratory stimulus (P < .001). There were no differences in the decrease in the MVC force and median percentage of integrated electromyography between 5 and 10 minutes of vibratory stimulus (P > .05). CONCLUSION: Prolonged vibratory stimulus (5 minutes) to the triceps brachii tendon appeared to have an inhibitory effect on MVC force and muscle activity. The present results suggest that prolonged vibratory stimulus could be an effective treatment capable of reducing muscle tonus of the triceps brachii.

4',6-Dimethoxyisoflavone-7-O-ß-D-glucopyranoside (wistin) is a peroxisome proliferator-activated receptor α (PPARα) agonist in mouse hepatocytes.

Peroxisome proliferator-activated receptors (PPARs) are ligand-dependent transcription factors that regulate lipid and glucose metabolism. PPARα mainly affects fatty acid metabolism, and its activation lowers lipid levels. PPARγ is involved in the regulation of adipogenesis, insulin sensitivity, energy balance, and lipid biosynthesis. We have previously reported that 4',6-dimethoxyisoflavone-7-O-ß-D-glucopyranoside (wistin) can activate PPARγ. The purpose of the present study is to investigate the PPARα agonist activity of wistin. Using a luciferase reporter assay system of PPARα in monkey COS7 kidney cells, we showed that wistin could activate PPARα (P < 0.01 at 10 µg/mL) in a dose-dependent manner. Moreover, the addition of wistin upregulated the expression of PPARα (P < 0.01 at 10 µg/mL) and PPARα target genes including carnitine palmitoyltransferase 1a (P < 0.05 at 10 µg/mL), acyl-CoA oxidase (P < 0.01 at 10 µg/mL), acyl-CoA synthase (P < 0.05 at 10 µg/mL), PPARγ coactivator 1α (P < 0.05 at 10 µg/mL), uncoupling protein 2 (P < 0.05 at 1 µg/mL), and uncoupling protein 3 (P < 0.05 at 10 µg/mL), which are genes involved in lipid efflux and energy expenditure, in mouse primary hepatocytes. Furthermore, wistin inhibited cellular triglyceride accumulation in hepatocytes (P < 0.05 at 10 µg/mL) in a dose-dependent manner. These results indicate that wistin could suppress lipid accumulation through PPARα activation. The action of wistin on PPARα could be of interest for the amelioration of lipid metabolic disorders. To the best of our knowledge, wistin is the first reported isoflavonoid O-glycoside with PPARα agonist activity.

Simultaneous determination of five polyether ionophores using liquid chromatography with one-step fluorescent derivatization.

We present a selective method for simultaneous determination of five polyether ionophores such as salinomycin (SAL), monensin (MON), narasin (NAR), semduramicin (SEM) and lasalocid (LAS) in aquatic samples using a liquid chromatography with one-step fluorescent derivatization of 2-(4-hydrazinocarbonyl-phenyl) 4,5-diphenylimidazole (HCPI) and 4-(4,5-diphenyl-1H-imidazol-2-yl) benzoyl chloride hydrochloride (DIB-Cl). Fluorescent one-step derivatization for SAL, MON, NAR and SEM using HCPI and for LAS using DIB-Cl was monitored by an LC/fluorescence detector (E(x), 340 nm; E(m), 465 nm). Chromatographic separation was performed on a TSK-GEL ODS-120T (4.6 × 150 mm, 3 µm) column using a mobile phase of 0.1% formic acid in acetonitrile and 0.5 mM ammonium formate in water (70/30, v/v). The limits of detections were 0.01 µg/mL (50 pg) for LAS, 0.05 µg/mL (250 pg) for SAL, NAR and SEM, and 0.1 µg/mL (500 pg) for MON, respectively. The recoveries for water samples were indicated to be the range of 79.6 ± 6.4 - 99.0 ± 4.4% with associated precision values (between-day for 3 days) for repeatability. Based on solid-phase extraction, the limit of quantitation values indicated 0.1 ng/mL for SAL, MON, NAR and SEM, and 0.01 ng/mL for LAS in water samples.