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Berlin Brandenburg

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  • 1
    Language: English
    In: Translational Research, 2010, Vol.156(2), pp.98-105
    Description: Activating V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) and V-raf murine sarcoma viral oncogene homolog B1 (BRAF) gene mutations are important predictive markers for antiepidermal growth factor receptor chemotherapy in colorectal cancer (CRC). However, a rapid and accurate assay for KRAS/BRAF mutation detection from routine pathological specimens is lacking in clinical practice. We applied the cycleave polymerase chain reaction (PCR) method to routine KRAS/BRAF genotyping of CRC patients at our institution from 2001 to 2009. The accuracy of cycleave PCR genotyping was shown by the high concordance with reverse transcriptase-PCR-coupled direct sequencing. KRAS gene mutations were analyzed successfully from small biopsy or cytology specimens. Although some surgical specimens could not be evaluated by cycleave PCR, corresponding biopsy specimens could be used instead. This PCR failure observed for some biopsy specimens may have been a result of the use of formalin fixation, as overfixation of surgical specimens by formalin impaired PCR amplification. In conclusion, cycleave PCR is practically applicable to KRAS/BRAF genotyping using small amounts of biopsied tumor cells. Care must be taken in the selection of pathological specimens for KRAS/BRAF testing.
    Keywords: Medicine
    ISSN: 1931-5244
    E-ISSN: 1878-1810
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  • 2
    In: PLoS ONE, 2016, Vol.11(10)
    Description: Phagocytes such as dendritic cells and macrophages, which are distributed in the small intestinal mucosa, play a crucial role in maintaining mucosal homeostasis by sampling the luminal gut microbiota. However, there is limited information regarding microbial uptake in a steady state. We investigated the composition of murine gut microbiota that is engulfed by phagocytes of specific subsets in the small intestinal lamina propria (SILP) and Peyer’s patches (PP). Analysis of bacterial 16S rRNA gene amplicon sequences revealed that: 1) all the phagocyte subsets in the SILP primarily engulfed Lactobacillus (the most abundant microbe in the small intestine), whereas CD11b hi and CD11b hi CD11c hi cell subsets in PP mostly engulfed segmented filamentous bacteria (indigenous bacteria in rodents that are reported to adhere to intestinal epithelial cells); and 2) among the Lactobacillus species engulfed by the SILP cell subsets, L . murinus was engulfed more frequently than L . taiwanensis , although both these Lactobacillus species were abundant in the small intestine under physiological conditions. These results suggest that small intestinal microbiota is selectively engulfed by phagocytes that localize in the adjacent intestinal mucosa in a steady state. These observations may provide insight into the crucial role of phagocytes in immune surveillance of the small intestinal mucosa.
    Keywords: Research Article ; Biology And Life Sciences ; Biology And Life Sciences ; Biology And Life Sciences ; Medicine And Health Sciences ; Biology And Life Sciences ; Medicine And Health Sciences ; Biology And Life Sciences ; Biology And Life Sciences ; Research And Analysis Methods ; Biology And Life Sciences ; Biology And Life Sciences ; Biology And Life Sciences ; Medicine And Health Sciences ; Biology And Life Sciences ; Biology And Life Sciences ; Medicine And Health Sciences ; Biology And Life Sciences ; Biology And Life Sciences ; Biology And Life Sciences
    E-ISSN: 1932-6203
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  • 3
    Language: English
    In: Biomaterials, April 2015, Vol.47, pp.62-71
    Description: The nanoscale structure-function relationship is a key determinant of bone toughness or micro-fragility. The loss of bone toughness during the aging process has been accepted based on empirical evidence, but this concept has not yet been fully supported by evidence at the material level. Here, we demonstrate a reduction in bone toughening mechanism in mimetic aged cortical bone obtained from deficient ( ) mice and assessed by dynamic mechanical analysis. The strain-rate nanoindentation tests showed enhanced stiffening of the wild-type calvarial bone and a large dimensional recovery during rapid loading following the constant displacement test. Such strain-dependent stiffening was likely associated with nanoscale dilatational bands and subsequent strain-energy transfer to the superior wild-type cross-linked collagen matrix network. The absence of dilatational bands formed by hydroxyapatite crystals and non-collagenous proteins in the bone samples likely diminished the intrinsic bone toughening mechanisms almost independent of viscoelastic behaviors. Such nanoscale structural alternations that occur during aging processes lead to crack propagation and result in overall bone fractures under large external stresses. In addition, dynamic mechanical analysis using instrumented nanoindentation was useful for the evaluation of bone mechanical properties in this pathological model of a genetic knockout mouse.
    Keywords: Bone ; Mechanical Properties ; Ageing ; Dynamic Mechanical Analysis ; Nanoindentation ; Medicine ; Engineering
    ISSN: 0142-9612
    ISSN: 20452322
    E-ISSN: 1878-5905
    E-ISSN: 20452322
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  • 4
    Language: English
    In: The Journal of Allergy and Clinical Immunology, April 2016, Vol.137(4), pp.1226-1235
    Description: The circadian clock temporally gates signaling through the high-affinity IgE receptor (FcεRI) in mast cells, thereby generating a marked day/night variation in allergic reactions. Thus manipulation of the molecular clock in mast cells might have therapeutic potential for IgE-mediated allergic reactions. We determined whether pharmacologically resetting the molecular clock in mast cells or basophils to times when FcεRI signaling was reduced (ie, when core circadian protein period 2 [PER2] is upregulated) resulted in suppression of IgE-mediated allergic reactions. We examined the effects of PF670462, a selective inhibitor of the key clock component casein kinase 1δ/ε, or glucocorticoid, both of which upregulated PER2 in mast cells, on IgE-mediated allergic reactions both and . PF670462 or corticosterone (or dexamethasone) suppressed IgE-mediated allergic reactions in mouse bone marrow–derived mast cells or basophils and passive cutaneous anaphylactic reactions in mice in association with increased PER2 levels in mast cells or basophils. PF670462 or dexamethasone also ameliorated allergic symptoms in a mouse model of allergic rhinitis and downregulated allergen-specific basophil reactivity in patients with allergic rhinitis. Pharmacologically resetting the molecular clock in mast cells or basophils to times when FcεRI signaling is reduced can inhibit IgE-mediated allergic reactions. The results suggest a new strategy for controlling IgE-mediated allergic diseases. Additionally, this study suggests a novel mechanism underlying the antiallergic actions of glucocorticoids that relies on the circadian clock, which might provide a novel insight into the pharmacology of this drug in allergic patients.
    Keywords: Circadian Clock ; Mast Cells ; Basophils ; Ige ; Allergy ; Medicine
    ISSN: 0091-6749
    E-ISSN: 1097-6825
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  • 5
    Language: English
    In: PLoS ONE, 01 January 2017, Vol.12(5), p.e0178123
    Description: Sarcopenia is an aging and disease-related syndrome characterized by progressive and generalized loss of skeletal muscle mass and strength, with the risk of frailty and poor quality of life. Sarcopenia is diagnosed by a decrease in skeletal muscle index (SMI) and reduction of either handgrip strength or gait speed. However, measurement of SMI is difficult for general physicians because it requires special equipment for bioelectrical impedance assay or dual-energy X-ray absorptiometry. The purpose of this study was, therefore, to explore a novel, simple diagnostic method of sarcopenia evaluation in patients with cardiovascular diseases (CVD).We retrospectively investigated 132 inpatients with CVD (age: 72±12 years, age range: 27-93 years, males: 61%) Binomial logistic regression and correlation analyses were used to assess the associations of sarcopenia with simple physical data and biomarkers, including muscle-related inflammation makers and nutritional markers.Sarcopenia was present in 29.5% of the study population. Serum concentrations of adiponectin and sialic acid were significantly higher in sarcopenic than non-sarcopenic CVD patients. Stepwise multivariate binomial logistic regression analysis revealed that adiponectin, sialic acid, sex, age, and body mass index were independent factors for sarcopenia detection. Sarcopenia index, obtained from the diagnostic regression formula for sarcopenia detection including the five independent factors, indicated a high accuracy in ROC curve analysis (sensitivity 94.9%, specificity 69.9%) and the cutoff value for sarcopenia detection was -1.6134. Sarcopenia index had a significant correlation with the conventional diagnostic parameters of sarcopenia.Our new sarcopenia index using simple parameters would be useful for diagnosing sarcopenia in CVD patients.
    Keywords: Sciences (General)
    E-ISSN: 1932-6203
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  • 6
    Language: English
    In: Nanomedicine: Nanotechnology, Biology, and Medicine, October 2013, Vol.9(7), pp.1036-1047
    Description: Bone morphogenetic protein-2 (BMP2) is among the most popular anabolic agents and substantially increase bone volume related to enhanced osteoblast differentiation. Here we demonstrate a remarkable deterioration in the nanomechanical properties of mineralized tissue induced from osteoblasts solely by the function of BMP2. Mineralized tissue of primary osteoblasts cultured with BMP2 shows molecular features of both bone and cartilage, but depletion of lysyl oxidase family members leads to poor nanomechanical properties of the mineralized tissue. Lysyl oxidase like-2 supplementation reinforces the inferior mineralized tissue induced from osteoblasts by BMP2 through intermolecular cross-linking of type II or type X collagen-rich extracellular matrix. This may also mimic a consolidation of bone fracture gaps, despite the fact that the distribution of the bone properties in such microenvironments has been poorly elucidated. These findings confirm the importance of testing newly induced bone down to the microscale and nanoscale in bone tissue engineering. Bone morphogenetic protein-2 is known to substantially increase bone volume related to enhanced osteoblast differentiation; however, this team of investigators report a remarkable deterioration in the nanomechanical properties of mineralized tissue induced from osteoblasts solely by the function of BMP2. Five indentation tests on an mineralize tissue. With the loading/partial unloading test, we can obtain constant hardness and elastic modulus on mineralized tissue along with indenter penetration.
    Keywords: Nanoindentation ; Raman Spectroscopy ; Osteoblasts ; Bone Morphogenetic Protein-2 ; Lysyl Oxidase Like-2 ; DNA Microarray ; Medicine
    ISSN: 1549-9634
    E-ISSN: 1549-9642
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  • 7
    Language: English
    In: Genome research, June 2003, Vol.13(6B), pp.1273-89
    Description: We report the construction of the mouse full-length cDNA encyclopedia,the most extensive view of a complex transcriptome,on the basis of preparing and sequencing 246 libraries. Before cloning,cDNAs were enriched in full-length by Cap-Trapper,and in most cases,aggressively subtracted/normalized. We have produced 1,442,236 successful 3'-end sequences clustered into 171,144 groups, from which 60,770 clones were fully sequenced cDNAs annotated in the FANTOM-2 annotation. We have also produced 547,149 5' end reads,which clustered into 124,258 groups. Altogether, these cDNAs were further grouped in 70,000 transcriptional units (TU),which represent the best coverage of a transcriptome so far. By monitoring the extent of normalization/subtraction, we define the tentative equivalent coverage (TEC),which was estimated to be equivalent to 〉12,000,000 ESTs derived from standard libraries. High coverage explains discrepancies between the very large numbers of clusters (and TUs) of this project,which also include non-protein-coding RNAs,and the lower gene number estimation of genome annotations. Altogether,5'-end clusters identify regions that are potential promoters for 8637 known genes and 5'-end clusters suggest the presence of almost 63,000 transcriptional starting points. An estimate of the frequency of polyadenylation signals suggests that at least half of the singletons in the EST set represent real mRNAs. Clones accounting for about half of the predicted TUs await further sequencing. The continued high-discovery rate suggests that the task of transcriptome discovery is not yet complete.
    Keywords: Cloning, Molecular ; Genome ; DNA, Complementary -- Genetics ; Transcription, Genetic -- Genetics
    ISSN: 1088-9051
    E-ISSN: 15495469
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  • 8
    In: Nanoscale, 2014, Vol.6(24), pp.14863-14871
    Description: While bone mineralization is considered to be responsible for its stiffness, bone durability partially associated with the time-dependent viscoelasticity of matrix proteins is still poorly elucidated. Here we demonstrate a novel mechanism of highly mineralized bone durability almost independent of inherent viscoelastic behaviour along with a protocol for measuring the mechanical properties of mineralized tissues. Strain-rate nanoindentation tests showed substantial stiffening of the highly mineralized calvarial bone, whereas large creep or stress relaxation was observed during constant load or displacement tests, respectively. Based on the lower viscoelasticity of the highly mineralized structure, such large time-dependent response appears to be associated with nanoscale dimensional recovery, rather than viscoelastic behaviour, implying the inverse namely strain-rate dependent dilatant behaviour. This dilatant expansion increased the indenter penetration resistance into the surface, enhancing instantaneous stiffness. The associated stiffening and higher effective elastic modulus were highly strain-rate dependent and more readily observed in more highly mineralized tissues such as the calvarial bone. Such strain-rate stiffening and consequent dimensional recovery may be vital responses of bone tissues against excessive deformation to maintain tissue integrity.
    Keywords: Steifigkeit ; Viskoelastizität ; Nanoeindruck (Härte) ; Mineralisierung ; Nanoeindringprüfung ; Spannungsrelaxation ; Belastungsprüfung (Mechanik) ; Protein ; Knochengewebe ; Elastizitätsmodul ; Nanotechnologie ; Knochen ; Viskoelastische Eigenschaft ; Engineering;
    ISSN: 2040-3364
    E-ISSN: 2040-3372
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  • 9
    Language: English
    In: International Journal of Oncology, January 2012, Vol.40(1), pp.93-98
    Description: Multiple mutations contribute to establish cancers. We have searched for potential oncogenes by screening cDNA libraries derived from gastric cancer cell lines, pancreatic cancer cell lines and glioma cell lines, using retrovirus-mediated expression cloning. Two types of interleukin-3 (IL-3)-dependent cell lines, Ba/F3 and HF6, were transduced with the cDNA libraries and several genes that render these cells factor-independent were identified including PIM-1, PIM-2, PIM-3, GADD45B and reproductive homeobox genes on the X chromosome gene F2 (RHOXF2). Although no mutation in these genes was found, these molecules were highly expressed in cancer cell lines and they may play important roles in cell transformation. Among them, we focused on a transcriptional repressor RHOXF2. Transduction of RHOXF2 rendered HF6 cells factor-independent, while knockdown of RHOXF2 inhibited growth of the HGC27 gastric cancer cell line which highly expresses RHOXF2. In addition, RHOXF2-transduced HF6 cells quickly induced leukemia when transplanted into sublethally irradiated mice. Moreover, RHOXF2 is highly expressed in some leukemia cell lines and a variety of human cancer samples including colon and lung cancers. Thus, these results indicate that RHOXF2 is involved in carcinogenesis.
    Keywords: Surgical Stapler;
    ISSN: 1019-6439
    E-ISSN: 17912423
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  • 10
    Language: English
    In: International journal of medicinal mushrooms, 2012, Vol.14(3), pp.257-69
    Description: Beta-glucan (BG) is a representative pathogen-associated molecular pattern (PAMP) produced by pathogenic fungi. SCG is a BG obtained from Sparassis crispa, which stimulates splenocytes in DBA/2 mice to produce cytokines, such as GM-CSF, IFN-γ, and TNF-α....
    Keywords: Actins -- Drug Effects ; Basidiomycota -- Chemistry ; Cytokines -- Metabolism ; Cytoskeleton -- Drug Effects ; Spleen -- Cytology ; Beta-Glucans -- Pharmacology
    ISSN: 1521-9437
    E-ISSN: 19404344
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