A rad50 germline mutation induces tumorigenesis and ataxia-telangiectasia phenotype in a transparent medaka model.

The MRE11A-RAD50-NBS1 complex activates the ataxia-telangiectasia mutated (ATM) pathway and plays a central role in genome homeostasis. The association of RAD50 mutations with disease remains unclear; hence, we adopted a medaka rad50 mutant to demonstrate the significance of RAD50 mutation in pathogenesis using the medaka as an experimental animal. A 2-base pair deletion in the rad50 gene was introduced into transparent STIII medaka using the CRISPR/Cas9 system. The mutant was analyzed histologically for tumorigenicity and hindbrain quality, as well as for swimming behavior, to compare with existing ATM-, MRE11A-, and NBS1-mutation-related pathology. Our results revealed that the medaka rad50 mutation concurrently reproduced tumorigenesis (8 out of 10 rad50Δ2/+ medaka), had a decrease in median survival time (65.7 ± 1.1 weeks in control vs. 54.2 ± 2.6 weeks in rad50Δ2/+ medaka, p = 0.001, Welch's t-test), exhibited semi-lethality in rad50Δ2/Δ2 medaka and most of the major ataxia-telangiectasia phenotypes, including ataxia (rheotaxis ability was lower in rad50Δ2/+ medaka than in the control, Mann-Whitney U test, p < 0.05), and telangiectasia (6 out of 10 rad50Δ2/+ medaka). The fish model may aid in further understanding the tumorigenesis and phenotype of ataxia-telangiectasia-related RAD50 germline mutations and in developing novel therapeutic strategies against RAD50 molecular disorders.

Can Individuals with Suboptimal Antibody Responses to Conventional Antiviral Vaccines Acquire Adequate Antibodies from SARS-CoV-2 mRNA Vaccination?

In Japan, healthcare workers (HCWs) are vaccinated against measles, rubella, chickenpox, mumps, and hepatitis B to prevent nosocomial infection; however, some do not produce sufficient antibodies ("suboptimal responders"). This study compared immune responses to a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 mRNA) vaccine among HCWs with normal and suboptimal responses to conventional vaccines. In this prospective cohort study, 50 HCWs received two doses of BNT162b2 mRNA vaccine 3 weeks apart. SARS-CoV-2 anti-spike antibodies were measured 11 times, starting before the first vaccination and ending 5 months after the second vaccination. Antibody titers of four suboptimal and 46 normal responders were compared. SARS-CoV-2 neutralizing antibody activity was measured twice in suboptimal responders, 1 week/1 month and 5 months after the second vaccination. The SARS-CoV-2 anti-spike antibody was detectable in the samples from suboptimal and normal responders at each timepoint after vaccination. Suboptimal responders exhibited SARS-CoV-2 neutralizing antibody activity 1 week/1 month as well as 5 months after the second vaccination; however, activity was slightly reduced at 5 months. Our findings show that suboptimal responders do acquire adequate SARS-CoV-2 anti-spike and SARS-CoV-2 neutralizing antibodies from vaccination to prevent SARS-CoV-2. SARS-CoV-2 mRNA vaccines should thus be recommended for both normal and suboptimal responders to conventional vaccines.

Sonographically Measured Adipose Tissue Thickness Correlates with Laboratory Test Abnormalities Reflecting Metabolic State in Elderly Women.

Background: Accumulation of adipose tissue progresses to metabolic diseases. Sonography is a convenient modality for measuring the thickness of adipose tissue. The present study aimed to clarify the site of adipose tissue thickness that correlated best with laboratory test values reflecting metabolic abnormalities. Methods: Subjects comprised 37 elderly women with metabolic diseases or an almost healthy state (median age, 71 years; interquartile range, 62-78 years). Abdominal visceral adipose tissue (VAT), subcutaneous adipose tissue, peritoneal adipose tissue, perirenal adipose tissue, and epicardial adipose tissue (EAT) thicknesses were measured. Correlations were evaluated between laboratory test values and these adipose tissue thicknesses. Results: VAT thickness measured at the level of the umbilicus correlated positively with values of triglycerides (TGs) (r = 0.593, P = 0.0009) and hemoglobin A1c (r = 0.490, P = 0.0081) and negatively with the value of high-density lipoprotein cholesterol (r = -0.521, P = 0.0045), even after adjusting for body mass index. Significant positive correlations were also found between EAT thickness and TGs (r = 0.542, P = 0.0029). Conclusions: Among the adipose tissue thicknesses measured at several sites by sonography, VAT thickness correlated most closely with laboratory test values representing metabolic abnormalities in elderly women.

Discrimination of well-differentiated liposarcoma from benign lipoma on sonography: an uncontrolled retrospective study.

PURPOSE: Well-differentiated liposarcoma, the most common subtype of liposarcoma, should be discriminated from benign lipoma. However, features on sonography for discriminating these two types of tumor have not been fully investigated. The present study was therefore aimed at clarifying differences in sonographic findings between well-differentiated liposarcoma and lipoma. METHODS: The study population comprised 23 cases of well-differentiated liposarcoma and 181 cases of lipoma. We investigated differences in sonographic appearance and pathological findings between the two types of tumor. RESULTS: Well-differentiated liposarcoma tended to develop more frequently in older patients and in the lower extremities including the gluteal region, compared with lipoma. Concerning sonographic findings, both tumors exhibited well-defined margins and heterogeneous internal echogenicity, including typical tiny striated hyperechoic lines. Well-differentiated liposarcoma was characterized by a higher frequency of the following findings compared with lipoma: (1) deep location, (2) irregular shape, (3) large diameter, (4) hyperechogenicity compared to surrounding tissue, and (5) presence of vascularity on Doppler sonography (p < 0.01 each). Notably, hyperechogenicity corresponded to the intermingled sclerosing component within the adipocytic component when sonographic findings were compared with those of pathology. CONCLUSION: The present study suggests that several sonographic findings including hyperechogenicity and presence of vascularity might be key features for discriminating well-differentiated liposarcoma from lipoma.

Sonographic Appearances of Liposarcoma: Correlations with Pathologic Subtypes.

Liposarcoma is the second most common malignant soft-tissue tumor. This entity is pathologically categorized into 4 subtypes: well-differentiated, myxoid, dedifferentiated and pleomorphic. Although features on magnetic resonance imaging and computed tomography for these 4 subtypes have been reported quite precisely, those on sonography have not been fully investigated. The present study was therefore aimed at clarifying the sonographic appearances of each liposarcoma subtype and assessing correlations with histopathology. The study population was made up of 35 cases, including 21 cases of well-differentiated liposarcoma, 6 cases of myxoid liposarcoma, 6 cases of dedifferentiated liposarcoma and 2 cases of pleomorphic liposarcoma. Compared with the other subtypes, well-differentiated liposarcoma was characterized by the high frequency of the following findings: isoechogenicity, tiny hyperechoic lines and hypovascularity (p < 0.01, in each). Myxoid liposarcomas were characterized by low echogenicity, intermingled with anechoic areas and moderate vascularity (p < 0.01, in each). Dedifferentiated liposarcomas showed a specific biphasic pattern of hyperechoic and hypoechoic areas and hypervascularity (p < 0.01, in each). Pleomorphic liposarcomas showed a specific gyrus-like mixture of hyperechoic and hypoechoic areas (p < 0.01). In conclusion, the present study revealed different characteristics of sonographic appearance among the 4 histopathologic subtypes of liposarcoma.

GLCCI1 is a novel protector against glucocorticoid-induced apoptosis in T cells.

Glucocorticoids (GCs) potently induce T-cell apoptosis in a GC receptor (GR)-dependent manner and are used to control lymphocyte function in clinical practice. However, its downstream pathways remain controversial. Here, we showed that GC-induced transcript 1 (GLCCI1) is a novel downstream molecule of the GC-GR cascade that acts as an antiapoptotic mediator in thymic T cells. GLCCI1 was highly phosphorylated and colocalized with microtubules in GLCCI1-transfected human embryonic kidney QBI293A cells. GR-dependent up-regulation of GLCCI1 was associated with GC-induced proapoptotic events in a cultured thymocyte cell line. However, GLCCI1 knockdown in a thymocyte cell line led to apoptosis. Consistently, transgenic mice overexpressing human GLCCI1 displayed enlarged thymi that consisted of larger numbers of thymocytes. Further molecular characterization showed that GLCCI1 bound to both dynein light chain LC8-type 1 (LC8) and its functional kinase, p21-protein activated kinase 1 (PAK1), thereby inhibiting the kinase activity of PAK1 toward LC8 phosphorylation, a crucial event in apoptotic signaling. GLCCI1 induction facilitated LC8 dimer formation and reduced Bim expression. Thus, GLCCI1 is a candidate factor involved in apoptosis regulation of thymic T cells.-Kiuchi, Z., Nishibori, Y., Kutsuna, S., Kotani, M., Hada, I., Kimura, T., Fukutomi, T., Fukuhara, D., Ito-Nitta, N., Kudo, A., Takata, T., Ishigaki, Y., Tomosugi, N., Tanaka, H., Matsushima, S., Ogasawara, S., Hirayama, Y., Takematsu, H., Yan, K. GLCCI1 is a novel protector against glucocorticoid-induced apoptosis in T cells.

KRAS Mutation as a Potential Prognostic Biomarker of Biliary Tract Cancers.

BACKGROUND: The aim of this study was to identify the unique molecular characteristics of biliary tract cancer (BTC) for the development of novel molecular-targeted therapies. MATERIALS AND METHODS: We performed mutational analysis of KRAS, BRAF, PIK3CA, and FBXW7 and immunohistochemical analysis of EGFR and TP53 in 63 Japanese patients with BTC and retrospectively evaluated the association between the molecular characteristics and clinicopathological features of BTC. RESULTS: KRAS mutations were identified in 9 (14%) of the 63 BTC patients; no mutations were detected within the analyzed regions of BRAF, PIK3CA, and FBXW7. EGFR overexpression was observed in 5 (8%) of the 63 tumors, while TP53 overexpression was observed in 48% (30/63) of the patients. Overall survival of patients with KRAS mutation was significantly shorter than that of patients with the wild-type KRAS gene (P = 0.005). By multivariate analysis incorporating molecular and clinicopathological features, KRAS mutations and lymph node metastasis were identified to be independently associated with shorter overall survival (KRAS, P = 0.004; lymph node metastasis, P = 0.015). CONCLUSIONS: Our data suggest that KRAS mutation is a poor prognosis predictive biomarker for the survival in BTC patients.

Relationship between Pericardial Adipose Tissue Thickness and Early Impairment of Left Ventricular Function, Both Evaluated on Echocardiography.

Epicardial adipose tissue (EAT) is metabolically bioactive, and accumulation of this tissue is related to early impairment of left ventricular (LV) systolic function as well as diastolic function. However, pericardial adi- pose tissue (PAT), located outside the EAT, has recently been demonstrated to be more closely associated with metabolic risk factors than EAT. The present study aimed to clarify whether PAT thickness is related to early impairment of LV function in a similar manner to EAT thickness, with both evaluated echocardio- graphically. Subjects were 49 women (mean age, 68ill years) composed of both patients with metabolic diseases and those with other diseases, and ejection fraction (EF) >55%. Systolic function was assessed by measuring EF, systolic mitral annular velocity (S'), and tissue mitral annular displacement percentage (TMAD%). Diastolic function was assessed by measuring early rapid filling wave velocity (E)/late filling wave velocity due to atrial contraction (A) ratio (E/A), peak early diastolic mitral annular velocity (e'), and E/e' ratio. Correlations between EAT or PAT thickness and LV systolic or diastolic function were assessed. EAT thickness correlated with S' and TMAD%(r=-0.399, p=0.005 and r=-0.570, p<0.001, respective- ly), but not with EF. However, PAT thickness was not correlated with any of these. EAT thickness corre- lated with E/A, e' and E/e'(r=-0.382, p=0.007; r=-0.493, p<0.001; and r=0.331, p=0.020, respective- ly). Again, PAT thickness was not correlated with any of these. PAT thickness appears unrelated to early impairment of LV function.

Relationship between epicardial adipose tissue thickness and early impairment of left ventricular systolic function in patients with preserved ejection fraction.

Epicardial adipose tissue (EAT) is metabolically bioactive fat. The present study aimed to clarify the relationship between EAT amount and early impairment of left ventricular (LV) systolic function in patients with preserved ejection fraction (EF), all evaluated echocardiographically. Participants comprised 62 elderly women (mean age ± standard deviation, 68 ± 11 years) with lifestyle-related diseases and EF ≥ 60 %. EAT amount was evaluated as thickness. Parameters suggesting early impairment of systolic function such as decreases in systolic mitral annular velocity (S') and tissue mitral annular displacement percentage (TMAD %) were evaluated along with EF. Correlations between EAT thickness and these LV systolic functions were assessed. Influences of various factors on the resultant significant relationships were also assessed. EAT thickness correlated inversely with S' and TMAD % (r = -0.402, p = 0.001 and r = -0.585, p < 0.001, respectively), but did not correlate with EF (r = 0.054, not significant). These significant relationships were maintained after considering factors such as body mass index, age, presence of lifestyle-related diseases and blood test results. A significant relationship existed between EAT amount and early impairment of LV systolic function in patients with preserved EF. Accumulation of EAT might contribute to the initial development of LV systolic dysfunction.

Whole-Genome Sequence of Mycobacterium kyorinense.

We report here the first draft genome sequence of Mycobacterium kyorinense, which was described in 2009 and exhibits significant pathogenicity to humans.

V843I, a lung cancer predisposing EGFR mutation, is responsible for resistance to EGFR tyrosine kinase inhibitors.

INTRODUCTION: We previously demonstrated that a family predisposed to lung cancer harbored a V843I substitution in the epidermal growth factor receptor (EGFR) protein. We report here the further characterization of this mutant EGFR protein in the context of tumorigenicity and resistance to tyrosine kinase inhibitors (TKIs) of EGFR activity. METHODS: Phosphorylation of EGFR and downstream signaling proteins of lung adenocarcinoma cell lines with EGFR mutations was assayed by flow cytometry. Susceptibility to TKIs of these cell lines, with or without suppression of mutant EGFR expression by small inhibitory RNA (siRNA), was investigated using a cellular viability assay. Furthermore, protein modeling was used to predict TKI binding to EGFR protein carrying the V843I mutation. RESULTS: Phosphorylation of EGFR and downstream signaling proteins was elevated upon transfection with an EGFR gene with the V843I. Although the cell line with V843I + L858R demonstrated resistance to EGFR-TKIs, the cells became susceptible to TKIs upon incubation with siRNA specific for the V843I allele. The structural analysis suggested that TKI binding to EGFR would be sterically hindered by Arg841 in the double-mutant (V843I + L858R) EGFR. CONCLUSIONS: The V843I mutation contributes to tumorigenesis by promoting phosphorylation of EGFR and its downstream signaling proteins. This mutation also appears to provide resistance to EGFR-TKIs through structural modification of EGFR. These features are comparable with those in EGFR T790M mutation, suggesting that cases with germ-line V843I or T790M mutations could be categorized as a class of familial lung cancer syndrome with resistance to EGFR-TKIs.

Carcinosarcoma, an atypical subset of gallbladder malignancies.

Carcinosarcoma represents an atypical subset of gallbladder malignancies, and sonographic imaging features have not yet been precisely defined. Previously reported cases have shown a heterogeneously echogenic solid mass protruding into and filling the gallbladder lumen. We present herein a case of carcinosarcoma and propose another finding suggestive of this tumor. The patient was a woman in her 70s. Abdominal sonography revealed that the gallbladder lumen was half-filled by a large mass (maximum diameter, 68 mm) showing heterogeneous echogenicity slightly higher than that of bile. However, despite the large size of the mass, gallbladder shape was well-preserved. Considering the findings on computed tomography, cholecystectomy was performed under a diagnosis of gallbladder malignancy. Pathological examination revealed two types of malignant histology: a sarcomatous element of malignant spindle cells and a carcinomatous element of adenocarcinoma tissue. Foci of malignant cartilage and bone areas were also found sporadically. Accompanied by immunohistochemical examination, the mass was diagnosed as carcinosarcoma. The present case showed somewhat different imaging findings from those of ordinary gallbladder carcinoma. Carcinosarcoma should be considered when a well-preserved shape of the gallbladder is recognized along with protrusion of a large heterogeneously echogenic mass into and filling the gallbladder lumen.

Primeiro isolamento de Mycobacterium kyorinense em espécime clínico no Brasil / First isolation of Mycobacterium kyorinense in a clinical specimen in Brazil

Os testes bioquímicos realizados, o seq³enciamento de diferentes alvos genéticos e a construþÒo de uma árvore concatenada, construída a través do método Neighbor-Joining, permitiram a identificaþÒo das cepas brasileiras como M. kyorinense. (AU)

Primeiro isolamento de Mycobacterium kyorinense em espécime clínico no Brasil / First isolation of Mycobacterium kyorinense in a clinical specimen in Brazil

Os testes bioquímicos realizados, o seqüenciamento de diferentes alvos genéticos e a construção de uma árvore concatenada, construída a través do método Neighbor-Joining, permitiram a identificação das cepas brasileiras como M. kyorinense.

First isolation of Mycobacterium kyorinense from clinical specimens in Brazil.

In this article, the first isolation of Mycobacterium kyorinense specimens in Brazil is described. M. kyorinense is a recently identified species, with a few strains reported only in Japan. The Brazilian isolates were initially identified as Mycobacterium celatum by PCR restriction enzyme pattern analysis (PRA) with hsp65. However, biochemical tests indicated the same profile of M. kyorinense and distinguished them from M. celatum and Mycobacterium branderi. The sequencing of the hsp65, rpoB, and 16S rRNA genes allowed the accurate identification of isolates as M. kyorinense.