Search for Gamma-Ray Spectral Lines from Dark Matter Annihilation up to 100 TeV toward the Galactic Center with MAGIC.

Linelike features in TeV γ rays constitute a "smoking gun" for TeV-scale particle dark matter and new physics. Probing the Galactic Center region with ground-based Cherenkov telescopes enables the search for TeV spectral features in immediate association with a dense dark matter reservoir at a sensitivity out of reach for satellite γ-ray detectors, and direct detection and collider experiments. We report on 223 hours of observations of the Galactic Center region with the MAGIC stereoscopic telescope system reaching γ-ray energies up to 100 TeV. We improved the sensitivity to spectral lines at high energies using large-zenith-angle observations and a novel background modeling method within a maximum-likelihood analysis in the energy domain. No linelike spectral feature is found in our analysis. Therefore, we constrain the cross section for dark matter annihilation into two photons to ⟨σv⟩≲5×10^{-28} cm^{3} s^{-1} at 1 TeV and ⟨σv⟩≲1×10^{-25} cm^{3} s^{-1} at 100 TeV, achieving the best limits to date for a dark matter mass above 20 TeV and a cuspy dark matter profile at the Galactic Center. Finally, we use the derived limits for both cuspy and cored dark matter profiles to constrain supersymmetric wino models.

Paratenon of the cruciate ligaments of the knee: a macroscopic and histological study of human foetuses.

BACKGROUND: The paratenon is a sheath-like connective tissue that allows the tendon to move with minimal friction. The careful removal of the paratenon along the cruciate ligaments is a critical step of knee surgery. Thus, orthopaedic surgeons and interventional radiologists consider the paratenon as a basic anatomical tissue along a ligament, not along a tendon. MATERIALS AND METHODS: We performed macroscopic and histological observations of cruciate ligament-associated paratenons in 43 human foetuses. RESULTS: This tissue usually had a thick armour-like appearance that was distant from the infrapatellar fat pad. The anterior cruciate ligament, rather than the posterior ligament, was deeply embedded in the paratenon. The paratenon contained abundant arteries and veins and, at and near the crossing between the cruciate ligaments, had a well-developed venous plexus. Notably, there were abundant fused veins in the paratenon venous plexus, and prenatal knee movements (especially rotation) seemed to restrict its blood supply, leading to the development of a large cavity by way of advancing fusion of veins in the degenerating plexus. This unique manner of cavitation likely expanded the joint cavity. CONCLUSIONS: Differences in knee movements in utero seemed to cause differences in the thickness of the paratenon among foetuses. New-borns might have limited knee flexion due to a mass-effect of the thick paratenon around the cruciate ligaments. A slight twisting or rotation at the knee may help to release the knee, because it can break the foetal paratenon and accelerate cavitation.

Fauna parasitária e relação parasito-hospedeiro de tambaquis criados na região do Baixo São Francisco, nordeste do Brasil / Parasitic fauna and parasite-host relationship of tambaqui reared in São Francisco river basin, Brazilian northeast

Este trabalho teve como objetivo avaliar a fauna parasitária de tambaquis na região do Baixo São Francisco-AL/SE-Brasil e correlacionar os índices de prevalência e intensidade média com fatores bióticos e abióticos. Foram coletados 252 espécimes para análise parasitológica de 10 pisciculturas. Os parasitos foram contabilizados, identificados, e determinaram-se os índices de prevalência e intensidade média, que foram correlacionados com fatores bióticos e abióticos. Dos peixes coletados, 65,5% estavam parasitados por pelo menos um táxon. Foram encontrados 10 táxons: Monogeneas, Ichthyophthirius multifiliis, tricodinídeos, Piscinoodinium pillulare, Ichthyobodo sp., Dolops carvalhoi, Lernaea cyprinacea, Procamallanus (Spirocamallanus) inopinatus, Henneguya sp. e Myxobolus sp. As maiores prevalências foram encontradas para Monogeneas (49,2%) e Myxobolus sp. (31,5%). Correlações negativas entre prevalência e fatores bióticos (peso e comprimento) foram observadas para Monogeneas (r2= -0,49; r2= -0,43), Myxobolus sp. (r²= -0,46; r²= -0,39) e Henneguya sp. (r²= -0,41; r²= -0,39). O fator abiótico temperatura apresentou correlação negativa com as prevalências de Lernaea cyprinacea (r= -0,39) e tricodinídeos (r= -0,33), enquanto a condutividade elétrica apresentou correlação positiva (r= 0,40) com a prevalência de tricodinídeos. Conclui-se que a fauna parasitária dos tambaquis cultivados na região do Baixo São Francisco é diversificada e com a carga parasitária dependente da qualidade de água e do estágio de desenvolvimento dos peixes.(AU)

This study investigated the parasitic fauna of tambaquis reared in lower Sao Francisco region-Al/SE-Brazil correlating parasitic indices to abiotic and biotic factors. A total of 252 specimens of tambaqui were collected in ten fish farms for parasitological analysis. The parasites were counted, identified and the parasitological indices were determined and correlated to biotic and abiotic factors. Of all collected fish, 65,5 % were parasitized by at least one taxon. Ten taxa were found: Monogeneans, Ichthyophthirius multifiliis, trichodinids Piscinoodinium pillulare, Ichthyobodo sp, Dolops carvalhoi, Lernaea cyprinacea, Procamallanus (Spirocamallanus) inopinatus, Henneguya sp. and Myxobolus sp. The higher prevalences were found to monogeneans (49.2%) and Myxobolus sp. (31.5%). Negative correlation of prevalence and biotic factor (weight and length) were observed to monogeneans (r 2 = -0.49, r 2 = -0.43), Myxobolus sp (r²= -0.46; r²= -0.39) and Henneguya sp (r²= -0.41; r²= -0.39). Abiotic factor of temperature presented a negative correlation to prevalence of Lernaea cyprinacea and trichodinids (r= -0.39 e r= -0.33, respectively) and the electric conductivity presented positive correlation to trichodinids (r= 0.40). It was concluded that parasitic fauna of tambaquis cultured in Lower São Francisco region is diversified and the parasitic load dependent on water parameters and fish growth.(AU)

SurePath® LBC improves the diagnostic accuracy of intrahepatic and hilar cholangiocarcinoma.

INTRODUCTION: The current study aimed to compare cytology using SurePath® (SP)-LBC and biliary tissue histology (BTH) for the diagnosis of biliary disease. METHODS: Between January 2014 and December 2016, 57 patients underwent endoscopic retrograde cholangiopancreatography for the diagnosis of biliary disease. Biliary cytological samples were processed using SP-LBC and subsequently BTH was performed. A final diagnosis was confirmed by surgery (23 malignant cases) and clinical follow-up (34 benign and malignant cases): 18 extrahepatic cholangiocarcinoma; 17 intrahepatic/hilar cholangiocarcinoma (intra/H-CC); eight other malignant disease; and 14 benign biliary disease. The diagnoses made using SP-LBC and BTH were classified into four categories: (1) benign; (2) indeterminate; (3) suspicious for malignancy/malignant; and (4) inadequate. In addition, diagnostic accuracy was compared between SP-LBC and BTH. RESULTS: Although 23% (13/57) of BTH samples were classified as inadequate, all SP-LBC cases were classified as adequate. Among 43 malignant cases, 11 normal, four indeterminate and 28 suspicious for malignancy/malignant were found using SP-LBC (26%, 9% and 65%, respectively), in contrast to 10 inadequate, nine normal, 10 indeterminate and 14 suspicious for malignancy/malignant observed using BTH (23%, 21%, 23%, and 33%, respectively). The identification of malignant cells was strikingly different between SP-LBC and BTH. Furthermore, limited to intra/H-CC, accuracy was significantly higher using SP-LBC than using BTH (P < .001). CONCLUSIONS: SP-LBC of the biliary tract is a useful and reliable method for diagnosing biliary malignant disease and has an advantage over BTH for detecting malignant cells and accurately diagnosing intra/H-CC.

Muscle layer thickness affects the peroral endoscopic myotomy procedure complexity.

Esophageal motility disorders can cause severe dysphagia, regurgitation, and/or noncardiac chest pain due to a lack of coordinated esophageal motility function. However, the clinical significance of esophageal muscle layer thickness remains unclear. The aims of this study are to elucidate the clinical significance of esophageal muscle layer thickness in patients with esophageal motility disorders who undergo peroral endoscopic myotomy (POEM), and to identify predictors of a longer POEM procedure time. Seventy-four consecutive patients with esophageal motility disorders who underwent POEM procedures at Kobe University Hospital from April 2015 to December 2016 were prospectively recruited into this study. First, we investigated the associations between the thickness of the esophageal muscular layer and clinical parameters. There were no significant differences, except in the POEM procedure time, between the patients with esophageal muscle layer thickness values of ≥1.5 mm (group A) and <1.5 mm (group B). However, the relative frequency of a longer POEM procedure time (≥78 min) was significantly higher in group A than in group B (66.7% vs. 19.5, P < 0.0001). Next, independent clinical factors that were related to longer POEM procedures were investigated. Multivariate logistic regression analysis with stepwise selection demonstrated that a thick esophageal muscle layer and the length of myotomy were an independent predictor of a longer POEM procedure (odds ratio: 13.9 and 12.0, respectively). Our results indicate that preoperative endoscopic ultrasonography evaluations can help to predict the technical complexity of POEM procedures.

A Combined test using both cell sediment and supernatant cell-free DNA in pleural effusion shows increased sensitivity in detecting activating EGFR mutation in lung cancer patients.

INTRODUCTION: The aim of this study was to examine whether a combined test using both cell sediment and supernatant cytology cell-free DNA (ccfDNA) is more useful in detecting EGFR mutation than using cell sediment DNA or supernatant ccfDNA alone in pleural effusion of lung cancer patients. METHODS: A total of 74 lung adenocarcinoma patients with paired samples between primary tumour and corresponding metastatic tumour with both cell sediment and supernatant ccfDNA of pleural effusion cytology were enrolled in this study. Cell sediment and supernatant ccfDNA were analysed separately for EGFR mutations by polymerase chain reaction. RESULTS: Out of 45 patients with mutant EGFR in primary tumours, EGFR mutations were detected in 23 cell sediments of corresponding metastases (sensitivity; 51.1%) and 20 supernatant ccfDNA corresponding metastases (sensitivity; 44.4%). By contrast, the combined test detected EGFR mutations in 27 corresponding metastases (sensitivity; 60.0%), and had a higher sensitivity than the cell sediment or the supernatant ccfDNA alone (P < .05). Out of 45 patients with mutant EGFR, 24, three and 18 were cytologically diagnosed as positive, atypical or negative, respectively. The detection rate in the combined test was highest (95.8%) in the positive group, and mutant EGFR was also detected in four of 18 samples (22.2%) in the negative group. CONCLUSIONS: A combined test using both cell sediment DNA and supernatant ccfDNA samples increases the concordance rate of EGFR mutations between primary tumour and corresponding metastases. Our findings indicate that supernatant ccfDNA is useful even in cases where the cytological diagnosis is negative.

Resposta hematológica do cascudo ornamental amazônico Peckoltia oligospila ao estresse de transporte / Hematological response of Amazonian ornamental Peckoltia oligospila subjected to stress by transportation

O objetivo deste trabalho foi avaliar as respostas hematológicas do acari-bola Peckoltia oligospila submetido ao estresse de transporte. Variações nos parâmetros de sangue foram analisadas às zero, seis, 24, 48, 72 e 96 horas após o transporte. Respostas ao estresse foram observadas entre zero e seis horas do transporte, mas a maioria dos parâmetros retornou aos valores basais em 24 horas. O tempo de zero hora (momento imediato após transporte) foi o mais crítico, com valores elevados de glicemia, eritrócitos e eritroblastos. Respostas secundárias tardias foram observadas para a proteína plasmática total, o volume corpuscular médio (VCM) e a hemoglobina corpuscular média (HCM) em seis horas após o transporte dos peixes, retornando aos valores basais após esse período. O número de leucócitos não sofreu alterações após o transporte. O estresse de transporte não comprometeu a fisiologia de P. oligospila, o que indica que esse peixe é resistente ao estresse se comparado com outras espécies. Porém, recomenda-se que não se realize qualquer outro procedimento estressante durante pelo menos 24 horas da recuperação dos peixes após transporte, para garantir a saúde e a sobrevivência dos animais transportados.(AU)

The objective of this work was to evaluate the hematological responses of bola-pleco (Peckoltia oligospila) undergoing the stress of transportation. Variations on blood parameters were analyzed at 0, 6, 24, 48, 72 and 96h after transportation. Responses to stress were detected from 0 to 6h after the transportation of fish, however, most parameters returned to baselines values within 24h of transportation. The moment of 0h was the most critical, presenting higher values of glycemia, erythroblasts and erythrocytes. Late secondary responses were observed to total plasmatic protein, mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) at 6h, returning to baselines values after this time. Leukocyte number was not affected by stress of transportation. The stress by transportation was not severe to influence the health of P. oligospila, indicating that fish is resistant to stress if compared to other species. However, we recommended no stressful procedures for at least 24 hours for recovery, in order to ensure health and survival of fish.(AU)

Efficacy of Ocimum gratissimum essential oil against the monogenean Cichlidogyrus tilapiae gill parasite of Nile tilapia / Eficácia do óleo essencial de Ocimum gratissimum contra o monogenea Cichlidogyrus tilapiae parasita de brânquias de tilápia-do-nilo

The phythotherapy is an alternative to use of chemotherapeutical agents against monogenean infection. This study evaluated the anthelmintic activity of essential oil Ocimum gratissimum against monogenean Cichlidogyrus tilapiae as well as its acute toxicity in tilapia juveniles. The mean lethal concentration (LC50) and different concentrations of the essential oil, both in vitro and in vivo assays (short and long-term baths) were assessed. The LC50 was 40.70mg.L-1 and in the in vitro assay this concentration showed 80% efficacy at the last two hours and in the in vivo assay 65.87% efficacy in long-term bath. However, it provoked morphological alterations on the gills such as hyperplasia and edema. The parasites exposure at the highest concentration (320mg.L-1) showed 100% mortality after 2h exposure in the in vitro assay, whereas in the in vivo assay, short-term baths of 5min for 3 consecutive days showed an efficacy of 87.71% without gills damage. These results demonstrate the anthelminthic activity of essential oil O. gratissimum and the safety concentration to use in Nile tilapia.(AU)

A fitoterapia é uma alternativa ao uso de agentes químicos contra infecções por parasitos monogenéticos. Este estudo avaliou a atividade anti-helmíntica do óleo essencial Ocimum gratissimum contra o monogenea Cichlidogyrus tilapiae , bem como sua toxicidade aguda e histopatologia em juvenis de tilápias. Foram avaliadas a concentração letal média (CL50) e diferentes concentrações de óleo essencial, tanto in vitro como in vivo (banho de curta e longa duração). A CL50 foi de 40,70mg.L-1; no ensaio in vitro, essa concentração apresentou 80% de eficácia, e no ensaio in vivo 65,87% de eficácia em banho de exposição crônica. No entanto, provocou alterações morfológicas nas brânquias, como hiperplasia e edema. A exposição dos parasitas na concentração mais elevada (320mg.L-1) mostrou 100% de mortalidade após duas horas de exposição no ensaio in vitro, enquanto no ensaio in vivo, em banho de curta duração de cinco minutos, durante três dias consecutivos, apresentou uma eficácia de 87,71%, sem danos às brânquias. Esses resultados demonstraram a atividade anti-helmíntica do óleo essencial de O. gratissimum e a concentração de segurança para uso na tilápia-do-nilo em banhos de curta duração.(AU)

Calreticulin mutant mice develop essential thrombocythemia that is ameliorated by the JAK inhibitor ruxolitinib.

Mutations of calreticulin (CALR) are detected in 25-30% of patients with essential thrombocythemia (ET) or primary myelofibrosis and cause frameshifts that result in proteins with a novel C-terminal. We demonstrate that CALR mutations activated signal transducer and activator of transcription 5 (STAT5) in 293T cells in the presence of thrombopoietin receptor (MPL). Human megakaryocytic CMK11-5 cells and erythroleukemic F-36P-MPL cells with knocked-in CALR mutations showed increased growth and acquisition of cytokine-independent growth, respectively, accompanied by STAT5 phosphorylation. Transgenic mice expressing a human CALR mutation with a 52 bp deletion (CALRdel52-transgenic mice (TG)) developed ET, with an increase in platelet count, but not hemoglobin level or white blood cell count, in association with an increase in bone marrow (BM) mature megakaryocytes. CALRdel52 BM cells did not drive away wild-type (WT) BM cells in in vivo competitive serial transplantation assays, suggesting that the self-renewal capacity of CALRdel52 hematopoietic stem cells (HSCs) was comparable to that of WT HSCs. Therapy with the Janus kinase (JAK) inhibitor ruxolitinib ameliorated the thrombocytosis in TG mice and attenuated the increase in number of BM megakaryocytes and HSCs. Taken together, our study provides a model showing that the C-terminal of mutant CALR activated JAK-STAT signaling specifically downstream of MPL and may have a central role in CALR-induced myeloproliferative neoplasms.

Inter-observer reproducibility of endometrial cytology by the Osaki Study Group method: utilising the Becton Dickinson SurePath™ liquid-based cytology.

OBJECTIVE: The purpose of the present study was to evaluate the reproducibility of the cytological diagnosis of endometrial lesions by the Osaki Study Group (OSG) method of new cytological diagnostic criteria using BD SurePath™ (SP)-liquid-based cytology (LBC). METHODS: This cytological classification using the OSG method consists of six categories: (i) normal endometrium (NE), (ii) endometrial glandular and stromal breakdown (EGBD), (iii) atypical endometrial cells, cannot exclude atypical endometrial hyperplasia or more (ATEC-A), (iv) adenocarcinoma including atypical endometrial hyperplasia or malignant tumour (Malignancy), (v) endometrial hyperplasia without atypia (EH) and (vi) atypical endometrial cells of undetermined significance (ATEC-US). For this study, a total 244 endometrial samplings were classified by two academic cytopathologists as follows: 147 NE cases , 36 EGBD cases , 47 Malignant cases, eight ATEC-A cases, two EH cases and four ATEC-US cases. To confirm the reproducibility of the diagnosis and to study the inter- and intra-observer agreement further, a second review round followed at 3-month intervals, which included three additional cytopathologists. RESULTS: The inter-observer agreement of NE classes improved progressively from 'good to fair' to 'excellent', with values increasing from 0.70 to 0.81. Both EGBD and Malignancy classes improved progressively from 'good to fair' to 'excellent', with values increasing from 0.62-0.63 to 0.84-0.95, respectively. The overall intra-observer agreement between the first and the second rounds was 'good to fair' to 'excellent', with values changing from 0.79 to 0.85. All kappa improvements were significant (P < 0.0001). CONCLUSION: In this study, it seemed that the use of the OSG method as the new diagnostic criteria for SP-LBC preparation, may be a valid method to improve the precision (reproducibility) of endometrial cytology.

Comparison of dynamic contrast-enhanced MRI parameters of breast lesions at 1.5 and 3.0 T: a pilot study.

OBJECTIVE: To compare dynamic contrast-enhanced (DCE) MRI parameters from scans of breast lesions at 1.5 and 3.0 T. METHODS: 11 patients underwent paired MRI examinations in both Philips 1.5 and 3.0 T systems (Best, Netherlands) using a standard clinical fat-suppressed, T1 weighted DCE-MRI protocol, with 70-76 s temporal resolution. Signal intensity vs time curves were fit with an empirical mathematical model to obtain semi-quantitative measures of uptake and washout rates as well as time-to-peak enhancement (TTP). Maximum percent enhancement and signal enhancement ratio (SER) were also measured for each lesion. Percent differences between parameters measured at the two field strengths were compared. RESULTS: TTP and SER parameters measured at 1.5 and 3.0 T were similar; with mean absolute differences of 19% and 22%, respectively. Maximum percent signal enhancement was significantly higher at 3 T than at 1.5 T (p = 0.006). Qualitative assessment showed that image quality was significantly higher at 3 T (p = 0.005). CONCLUSION: Our results suggest that TTP and SER are more robust to field strength change than other measured kinetic parameters, and therefore measurements of these parameters can be more easily standardized than measurements of other parameters derived from DCE-MRI. Semi-quantitative measures of overall kinetic curve shape showed higher reproducibility than do discrete classification of kinetic curve early and delayed phases in a majority of the cases studied. ADVANCES IN KNOWLEDGE: Qualitative measures of curve shape are not consistent across field strength even when acquisition parameters are standardized. Quantitative measures of overall kinetic curve shape, by contrast, have higher reproducibility.

Insulin-like growth factor-1 regulates the expression of luteinizing hormone receptor and steroid production in bovine granulosa cells.

Luteinizing hormone LH plays important roles in follicular maturation and ovulation. The effects of LH are mediated by LH receptor (LHR) in the ovary. However, the factors that regulate the expression of LHR in bovine granulosa cells (GCs) are not well known. Insulin-like growth factor-1 (IGF-1) is known to play a key role in the acquisition and maintenance of functional dominance. To better understand the roles of LHR expression and IGF-1, we conducted three experiments to determine (i) mRNA expression of LHR in the GCs of developing follicles, (ii) the effects of IGF-1 on LHR mRNA expression in cultured GCs and (iii) the effects of IGF-1 on estradiol (E2), progesterone (P4) and androstenedione (A4) production by non-luteinized GCs. In experiment 1, small follicles (<6 mm Ø) expressed lower levels of LHR than mid-sized follicles (6-8 mm Ø) and large follicles (≥9 mm Ø) expressed the highest levels of LHR mRNA (p < 0.05). In experiment 2, IGF-1 (1 and 100 ng/ml) increased (p < 0.05) the expression of LHR mRNA in GCs from small and large follicles. In experiment 3, IGF-1 (0.1-100 ng/ml) increased A4 and E2 in GCs from both small and large follicles but increased P4 only in large follicles. IGF-1 in combination with LH (0.1 and 1 ng/ml) increased P4 and A4 in large follicles, and increased E2 and A4 in GCs of small follicles. These findings strongly support the concept that IGF-1 upregulates LHR mRNA expression as well as A4 and E2 production in GCs and that IGF-1 is required for determining which follicle becomes dominant and acquires ovulatory capacity.

Cytological nuclear atypia classification can predict prognosis in patients with endometrial cancer.

OBJECTIVE: Endometrial cancer is one of the leading causes of malignancy in females. Nuclear findings are important for patients with cancer, and can provide valuable information to treating oncologists. We investigated whether nuclear findings were a useful prognostic factor in patients with endometrial cancer. METHOD: We investigated 71 cases of endometrial carcinoma with paired histology and cytology at Kurume University Hospital. We classified endometrial endometrioid adenocarcinoma (EEC) G1 and G2 as type I carcinomas, and uterine papillary serous carcinoma (UPSC), clear cell carcinoma (CC) and EEC G3 as type II carcinomas. For the establishment of the cytological nuclear atypia classification, we examined the following nuclear factors on the cytological smears: mitotic figures, prominent nucleoli, nuclear area and anisonucleosis. RESULTS: There was a significant difference in mitotic figures (P < 0.001) and anisonucleosis (P = 0.026) in cytological smears between type I and type II carcinomas. Based on these findings, we categorized cytological nuclear atypia into three groups, nuclear atypia-1 (57.7%), nuclear atypia-2 (19.7%) and nuclear atypia-3 (22.5%), and this classification system correlated well with prognosis in patients with endometrial cancer (P < 0.001). Furthermore, this classification system was able to extract patients with a good prognosis from those with high-grade carcinomas, such as UPSC+CC+EEC G3, and patients with a poor prognosis from those with EEC G1. CONCLUSIONS: Our system of cytological nuclear atypia classification based on endometrial cytology can predict patient prognosis. Cytological nuclear atypia classification and histological typing may be useful for the treatment and follow-up of patients with endometrial cancer, and should be routinely incorporated into cytological reports.

Human microRNA hsa-miR-1231 suppresses hepatitis B virus replication by targeting core mRNA.

Pathogen-specific miRNA profiles might reveal potential new avenues for therapy. To identify miRNAs directly associated with hepatitis B virus (HBV) in hepatocytes, we performed a miRNA array analysis using urokinase-type plasminogen activator (uPA)-severe combined immunodeficiency (SCID) mice where the livers were highly repopulated with human hepatocytes and human immune cells are absent. Mice were inoculated with HBV-infected patient serum samples. Eight weeks after HBV infection, human hepatocytes were collected from liver tissues, and miRNAs were analysed using the Toray 3D array system. The effect of miRNAs on HBV replication was analysed using HBV-transfected HepG2 cells. Four miRNAs, hsa-miR-486-3p, hsa-miR-1908, hsa-miR-675 and hsa-miR-1231 were upregulated in mouse and human livers with HBV infection. These miRNAs were associated with immune response pathways such as inflammation mediated by chemokine and cytokine signalling. Of these miRNAs, hsa-miR-1231, which showed high homology with HBV core and HBx sequences, was most highly upregulated. In HBV-transfected HepG2 cells, overexpression of hsa-miR-1231 resulted in suppression of HBV replication with HBV core reduction. In conclusion, a novel interaction between hsa-miR-1231 and HBV replication was identified. This interaction might be useful in developing new therapeutic strategies against HBV.

TET2 is essential for survival and hematopoietic stem cell homeostasis.

Ten-Eleven-Translocation 2 (TET2) is an enzyme that catalyzes the conversion of 5-methylcytosine into 5-hydroxymethylcytosine (5-hmC) and thereby alters the epigenetic state of DNA; somatic loss-of-function mutations of TET2 are frequently observed in patients with diverse myeloid malignancies. To study the function of TET2 in vivo, we analyzed Ayu17-449 (TET2(trap)) mice, in which a gene trap insertion in intron 2 of TET2 reduces TET2 mRNA levels to about 20% of that found in wild-type (WT) mice. TET2(trap/trap) mice were born at Mendelian frequency but died at a high rate by postnatal day 3, indicating the essential role of TET2 for survival. Loss of TET2 results in an increase in the number of hematopoietic stem cells (HSCs)/progenitors in the fetal liver, and TET2(trap/trap) HSCs exhibit an increased self-renewal ability in vivo. In competitive transplantation assays, TET2(trap/trap) HSCs possess a competitive growth advantage over WT HSCs. These data indicate that TET2 has a critical role in survival and HSC homeostasis.

Host-derived MMP-13 exhibits a protective role in lung metastasis of melanoma cells by local endostatin production.

BACKGROUND: Although matrix metalloproteinases (MMPs) are implicated in tumourigenesis and cancer progression, the role of MMP-13 in melanoma cell metastases is poorly understood. METHODS: Lung metastases of mouse melanoma B16BL6 cells were analysed in MMP-13 knockout (KO) and wild-type (WT) mice after intravenous injection. The mRNA and protein expression of MMP-13 in lung tissues was analysed by RT-PCR, real-time PCR, immunoblotting and immunohistochemistry. The expression of SDF-1α, CXCR4 and endostatin, and effects of endostatin to cultured melanoma cells and lung metastases were also studied. RESULTS: Lung metastases of B16BL6 cells were significantly higher by 2.5-5.7-fold in MMP-13 KO mice than in WT mice. The expression of MMP-13 in WT mouse lung tissue was stimulated on day 1 after intravenous injection of the melanoma cells and MMP-13 was immunolocalised to vascular endothelial cells in the lungs. Endostatin formation, but not degradation of SDF-1α, in the lung tissue was associated with reduced lung metastasis in WT mice. Endostatin significantly inhibited migration of B16BL6 cells in monolayer wounding assay and remarkably suppressed Matrigel invasion and transendothelial invasion of the cells. In addition, lung metastases of melanoma cells in MMP-13 KO mice were reduced by intraperitoneal administration of endostatin. CONCLUSION: Our results suggest that MMP-13 is overproduced by endothelial cells in the lungs with melanoma cells and has a protective role in lung metastasis by local generation of endostatin.

Characterizing early contrast uptake of ductal carcinoma in situ with high temporal resolution dynamic contrast-enhanced MRI of the breast: a pilot study.

Improvements in the reliable diagnosis of preinvasive ductal carcinoma in situ (DCIS) by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) are needed. In this study, we present a new characterization of early contrast kinetics of DCIS using high temporal resolution (HiT) DCE-MRI and compare it with other breast lesions and normal parenchyma. Forty patients with mammographic calcifications suspicious for DCIS were selected for HiT imaging using T(1)-weighted DCE-MRI with ∼7 s temporal resolution for 90 s post-contrast injection. Pixel-based and whole-lesion kinetic curves were fit to an empirical mathematical model (EMM) and several secondary kinetic parameters derived. Using the EMM parameterized and fitted concentration time curve for subsequent analysis allowed for calculation of kinetic parameters that were less susceptible to fluctuations due to noise. The parameters' initial area under the curve (iAUC) and contrast concentration at 1 min (C(1 min)) provided the highest diagnostic accuracy in the task of distinguishing pathologically proven DCIS from normal tissue. There was a trend for DCIS lesions with solid architectural pattern to exhibit a negative slope at 1 min (i.e. increased washout rate) compared to those with a cribriform pattern (p < 0.04). This pilot study demonstrates the feasibility of quantitative analysis of early contrast kinetics at high temporal resolution and points to the potential for such an analysis to improve the characterization of DCIS.

Cellular localization of genes and proteins for tumor necrosis factor-α (TNF), TNF receptor types I and II in bovine endometrium.

To determine which cell types produce tumor necrosis factor-α (TNF) and its receptors (TNFRI and TNFRII) in bovine endometrium, we investigated the expression and cellular localization of their mRNAs and proteins. TNF transcripts and proteins were co-localized in endometrial epithelial cells, glandular epithelial cells and endothelial cells of microvessels but not in the stromal cells. TNF protein was detected in the lysate and the cultured media of epithelial cells, but was only weakly detected in the stromal cells. Both TNFRI (TNFRSF1A) and TNFRII (TNFRSF1B) transcripts were expressed in the epithelial cells, glandular epithelial cells and the stromal cells, whereas their proteins were weakly expressed in the stroma. TNF mRNA and protein expressions in the cultured epithelial cells were increased by TNF and interleukin-1α, and the TNFRII mRNA expressions were stimulated by oxytocin. Together, TNF secreted by the endometrial cells may locally play a role in regulating uterine function throughout the estrous cycle.