text:
Supplementary MaterialsDescription of extra supplementary items 42003_2019_507_MOESM1_ESM. and heterobifunctional glycan-centered ligands to give a quantitative high-throughput method for screening glycan libraries against glycan-binding and glycan-processing proteins. Software of the assay to human being (siglec-2), plant (and lectins) and bacterial (cholera toxin, and family 51 carbohydrate binding module) proteins allowed for the identification of ligands with affinities (lectin (SNA), which is specific for 2-6-linked sialosides10, a fragment of family 51 carbohydrate binding module (CBM51), which recognizes A and B type 2 and 6 blood group antigens11, and a soluble form of human being siglec-2 (CD22), which binds to 2-6 sialosides12,13, served as positive settings to validate CUPRA. We also investigated the lectins (MAA), a mixture of leucoagglutinin (MAL) and hemagglutinin (MAH). The preferred binding order Vitexin motif of MAH is definitely reported to become Neu5Ac2-3Gal1-3GalNAc (Neu5Ac 5Cmeasured by CUPRA screening of the 66-component library against S4 (50?M). As expected, the average for this bad control is close to 1.0 (0.99??0.03). In contrast, CUPRA screening against the positive controls produced decreases in for one or more of the library components. CUPRA screening of the library against CTB5 (10?M) identified the structurally-related GA01 (GM1) and GA02 (fucosyl-GM1) as ligands (Figs.?2b, c), while ABH11, ABH12, ABH13, ABH15, ABH19 and ABH20, which contain A and B order Vitexin type 2 tetra- and pentasaccharides, were identified as ligands of CBM51 (Supplementary Fig.?3a). HMO08 (composed of 6-sialyllactose), the only compound in the library with 2-6-linked Neu5Ac, was identified as a ligand of SNA (Supplementary Fig.?3b). Application of CUPRA to human CD22, part of an important class of Neu5Ac-binding GBPs implicated in both innate and adaptive immunity19, correctly identified HMO08 as a ligand (Supplementary Fig.?4a). CUPRA screening of MAA at high concentration (50?M) identified HMO03 and GA06 as ligands. Both of these ligands contain the Neu5Ac2-3Gal1-4Glc structure, although, in the case of GA06, it is capped by 2-8-linked Neu5Ac. The absence of detectable binding of MAA to GA03, which contains Neu5Ac2-3Gal1-3GlcNAc, the preferred binding motif order Vitexin of MAH, suggests a much lower affinity for this monovalent interaction. Open in a separate window Fig. 2 CUPRA enables screening of glycan libraries against GBPs and quantifying substrate specificity of CAZymes. a Library screening results for the negative control, streptavidin (50?M). The standard deviation in individual values determined from four measurements is 0.03. b, c Glycan library screening against the positive control cholera toxin B subunit homopentamer (CTB5) at initial concentrations of 2?M and 10?M, respectively. Depleted library components are shown by red bars. d, e Time-dependent substrate fractional abundance measured by CUPRA for 13 Neu5Ac-containing OSmod Mmp2 in the presence of human neuraminidase NEU2 and NEU3, respectively, at pH 7 and 25?C. Error bars represent standard deviations calculated for (CTB5, 58,020?Da, purity? ?95%), agglutinin (MAA, 130?kDa, purity? ?85%) and (SNA, 140?kDa, purity? ?90%) lectins were purchased from Sigma-Aldrich (Canada). A gene fragment encoding a family 51 carbohydrate-binding module (CBM51, order Vitexin MW 20 735?Da, purity? ?95%) was recombinantly produced in and purified as described elsewhere27. Residues 1C332 of human Siglec-2 (MW 140?kDa, purity? ?95%) were cloned in frame with human IgG1 Fc and a C-terminal His6, as described previously28. This chimeric construct, in the vector, was stably transfected into Lec-1 cell line through system under selection with 0.5?mg?mL?1 hygromycin-B for ~2 weeks. For expression, cells were grown in T-175 flasks for 12 d after reaching confluency, in 50?mL of DMEM-F12 media containing 10% FBS, 0.5% penicillin-streptomycin, and 1% HEPES. The protein supernatant was harvested, centrifuged (300 rcf, 10?min) and sterilized through a 0.5?M filter for storage at 4?C. For purification, 130?mL of the supernatant was loaded at 1 mL mL-1 onto a 1?mL Histrap Excel column (GE healthcare) equilibrated with 20?mM sodium phosphate, 0.5?M NaCl at pH 7.4. After loading, the column was washed with 15?mL of 30?mM imidazole in 20?mM sodium phosphate, 0.5?M NaCl at pH 7.4, then eluted with 500?mM imidazole in 20?mM sodium phosphate, 0.5?M NaCl, at pH 7.4. Fractions containing protein were diluted 10-fold in 20?mM phosphate buffer at pH 7.0. The order Vitexin diluted fractions were loaded onto a Protein-G column (GE healthcare) equilibrated with 20?mM phosphate buffer. The loaded protein on the column was washed with 15?mL of 20?mM phosphate buffer (pH 7.0) and eluted with 100?mM glycine solution at pH 2.7 via syringe and neutralized with 40-50?L of 1 1?M Tris buffer at pH 9.0 per 1?mL fraction. Fractions containing protein were dialyzed into 2?L of 200?mM ammonium acetate three times. Finally, the.
Category Archives: sGC
Casein kinase 1 is a group of ubiquitous serine/threonine kinases that
Casein kinase 1 is a group of ubiquitous serine/threonine kinases that are involved in normal cellular functions and several pathological conditions, such as DNA repair, cell cycle progression, cytokinesis, differentiation, and apoptosis. cell division and tumor growth in human pancreatic adenocarcinoma and salivary gland cancer by phosphorylating key proteins in the Wnt signaling pathway [15C18]. Changes in CK1? activity and expression as well as the occurrence of mutations within the coding region of CK1? have already been reported in a variety of malignancies including mammary ductal carcinoma, ovary tumor, and breast cancers [19,20]. Furthermore, CK1 epsilon substances could be utilized as potential restorative targets in the treating digestive malignancies [20]. However, the role of manifestation levels like a prognostic biomarker in dental cancer is not investigated. In this scholarly study, the expression was examined by us of CK1? in a big collection of dental cancer tissue examples to assess if CK1? might serve mainly because a predictor of results. We attemptedto assess associations between CK1 also? manifestation and clinicopathological guidelines of dental cancer individuals and the partnership to results. 2.?Discussion and Results 2.1. Individual Characteristics A complete of 195 individuals, including 159 males and 36 ladies were analyzed with this retrospective research. The individuals characteristics, like the individuals sex, age, cancers stage, lymph node position, histological grade, tumor position, smoking practices, betel nut nibbling habits, and general survival are detailed in Table 1. The mean age group of the 195 individuals was 55.9 years (a variety of 31C88 years). There have been 60 individuals with stage I, 56 with stage II, 38 with stage III, and 41 Faslodex irreversible inhibition with stage IV dental tumors. Twenty-seven tumors had been well-differentiated, 161 were Faslodex irreversible inhibition differentiated moderately, and only one 1 was differentiated poorly. In this research, we examined low T-stage (T1, T2 and T3) dental cancer individuals; there have been 80 instances Faslodex irreversible inhibition with T1 position, 93 with T2 position, and 22 with T3 position. The mean of general success was 4.1 years as well as the median survival time was 3.9 years. The entire survival period ranged from 0.1 to 9.6 years. Adjuvant therapy was given according to specific considerations. Desk 1. Patient characteristics. = 195 0.05). Open in a separate window Open in a separate window Physique 1. CK1? and CK1 immunoreactivity and CK1? kinase activity in non-tumor oral squamous mucosa and SCC. (A) Strong cytoplasmic CK1? expression in SCC (score 2+); (B) Strong cytoplasmic CK1 expression in SCC (score 2+); (C) Weak cytoplasmic CK1? expression in SCC (score 1+); (D) Weak cytoplasmic CK1 expression in SCC (score 1+); (E) Unfavorable CK1? expression in SCC (score 0); (F) Unfavorable CK1 expression in SCC (score 0); (G,H) Strong cytoplasmic CK1? expression in the non-tumor part than that of tumor part. Circle indicated the tumor part. Frame indicated the non-tumor part; and (I) Lower CK1? kinase activity in the tumor part then that of non-tumor part. * Significant differences from control values with Zfp264 0.05. We divided the CK1? and CK1 immunohistological stains into positive (1+/2+) and unfavorable (0) stain subgroups. The correlation between the expression level of CK1? and clinical parameters is usually summarized in Table 2. The chi-square analyses for the clinicopathologic characteristics of 195 patients with OSCC in relation to cytoplasmic CK1? expression showed that a unfavorable CK1? expression was considerably correlated with a short four-year survival (= 0.002). However, no significant association of CK1 expression was achieved with gender, age and the clinical parameters. Furthermore, we also test CK1? expression in normal epithelial cells and four different OSCC cells lines. Western blot analysis indicated CK1? expression in all oral cancer cells as well as normal squamous epithelial cells (Physique 2). The expression of CK1? in normal epithelial cells (SG cells) was higher than that in OECM1 and TW206 oral cancer cells (Physique 2). Open in a separate window Physique 2..
We present a case of leptomeningeal metastases in a 30-year-old man
We present a case of leptomeningeal metastases in a 30-year-old man with an extragonadal germ cell tumor. such patients is not yet acceptable. Delamanid irreversible inhibition Intrathecal metastasis (meningeal dissemination) of GCT is usually rare, and no effective treatment has been established. We present a case that temporarily responded to chemotherapy and discuss the possible treatment options in such cases. Case Statement A 30-year-old man with an extragonadal GCT of mediastinal origin complained of right homonymous hemianopia and was referred to Shinshu University Hospital for the treatment of the brain metastasis. About 4 months earlier, the patient experienced received induction chemotherapy with three courses of BEP (bleomycin, etoposide, and cisplatin) for multiple lung metastases in another hospital. The brain lesion had produced rapidly (Fig. 1), and serum tumor markers were elevated, including lactate dehydrogenase (LDH) at 447 U/l (normal, 118C236), fetoprotein (AFP) at 480 ng/ml (normal Q10), and human chorionic gonadotropin (HCG) at 629 IU/l (normal Q1.0). The patient had immediate neurosurgery to remove the tumor. On pathology, the tumor was diagnosed as metastatic embryonal carcinoma and yolk sac tumor (Fig. 2). The patient also received whole brain radiation therapy (WBRT) (total 30 Gy/10 fractions) and adjuvant chemotherapy consisting of two courses of VIP therapy (etoposide, ifosfamide, and cisplatin). All tumor markers normalized. The residual lung tumors were resected, and no tumor remnant was seen on pathology. Open in a separate window Physique 1 Brain MRI. The solitary metastatic lesion was shown, which was 4.5 cm 3 cm in diameter, occupying the left occipital lobe (before operation). Open in a separate windows Physique 2 H&E staining showed solid and tubular tumor cells with large, bizarre nuclei, coexisted with hemorrhage (magnification, 100). Immunohistochemical staining showed tumor cells to be positive with CD30 and AFP that suggested the embryonal carcinoma with yolk sac tumor component, and partially positive with HCG, suggesting syncytiotrophoblastic cells. One month after lung surgery, the patient developed lumbago and a gait disturbance. On magnetic resonance imaging (MRI), multiple leptomeningeal metastases with no brain recurrence were seen (Fig. 3A). Salvage chemotherapy, consisting of combination irinotecan and nedaplatin therapy, was given. After four courses of therapy, the leptomeningeal lesions disappeared (Fig. 3B). However, the serum HCG elevated again, and recurrent cerebral ventricular and meningeal lesions appeared within a short period of time. Despite the use of additional salvage chemotherapy including three courses of TIN therapy (paclitaxel, ifosfamide, and nedaplatin), in combination with the peripheral blood stem cell transplantation, the patient died due to disease progression 12 months after the initial brain surgery. Open in a separate window Physique 3 MRI of the spinal cord before and after salvage chemotherapy. 3A-Well-enhanced, multiple leptomeningeal metastases were detected at the 2nd and 12th thoracic cord levels, as well as at the 1st sacral cord level (Th2, Th12, and S1) (white arrows). 3B-Total remission of the leptomeningeal metastases was exhibited after salvage chemotherapy. Conversation Brain metastasis from malignant GCT occurs in 1%C3% of patients (Bokemeyer et al. 1997; Fossa et Delamanid irreversible inhibition al. 1999). Multidisciplinary treatment, including chemotherapy, irradiation, Delamanid irreversible inhibition and surgery, is required to treat brain GCT metastases. The prognosis of patients with brain Delamanid irreversible inhibition metastasis is usually poor, but it is usually relatively better in patients with a solitary (isolated) metastasis and in those with an initial metastasis compared to patients with multiple lesions or a relapse (Bokemeyer et al. 1997; Fossa et al. 1999; Mahalati et al. 1999; Lutterbach et al. 2002). Fossa et al. reported that this Pdgfa 5-year survival rate of patients with an initial metastasis was 45%, Delamanid irreversible inhibition but that of patients with recurrence after induction chemotherapy was 12% (Fossa et al. 1999). According to guidelines of the American National Malignancy Institute (NCI) and the European Urological Association, the standard treatment for GCT brain metastasis is usually chemotherapy in conjunction with WBRT (NCI, 2008; Albers et al. 2005). On the other hand, Salvati et al. recommended that, if the brain tumor is usually resectable, aggressive surgical treatment followed by WBRT and/or adjuvant chemotherapy should be given (Salvati et al. 2006). The incidence of leptomeningeal metastasis of main central nervous system (CNS) tumors has been reported to range from 7% to 27%, but extra-CNS GCT metastasis is usually rare (Engelhard et al. 2005). There has been only one statement in the last 10 years (Miranda et al. 2005). The etiology of.
Supplementary Materials [Supplemental material] molcellb_25_20_8779__index. to the RNA, a trimethylguanosine (TMG)
Supplementary Materials [Supplemental material] molcellb_25_20_8779__index. to the RNA, a trimethylguanosine (TMG) cap (m2,2,7GpppN), compared to the typical m7GpppN eukaryotic cap (30, 37, 52, 55). However, not all cellular mRNAs in splicing on Rabbit Polyclonal to KCY mRNA metabolism and to understand how mRNA metabolism and cap-interacting proteins have accommodated these two LY2228820 price RNA populations. To address these questions, we have chosen to use two nematodes as model systems, and embryos, we have developed both in vitro translation and decay systems as well as biolistic methods to evaluate the role of the spliced leader sequence and TMG cap on mRNA translation and decay (6, 12, 27). Using the in vitro decay system, we recently demonstrated that the predominant general pathway of mRNA decay is 3-to-5 exonucleolytic decay followed by scavenger hydrolysis of the resulting mRNA cap (6). 5-to-3 decay also occurs in the extracts but is 15-fold less active than the 3-to-5 decay LY2228820 price pathway. Both pathways in vitro are capable of hydrolysis of both the m7GpppN cap and the m2,2,7GpppN cap derived from the spliced leader. We have also shown that the recombinant scavenger enzyme DcpS can hydrolyze both the m7GpppN and m2,2,7GpppN caps. In LY2228820 price contrast, human DcpS is not LY2228820 price capable of hydrolyzing m2,2,7GpppN caps, and the enzyme substrate requirements differ from those of the human enzyme (6). To extend these studies on mRNA turnover and decapping in nematodes, we have now cloned, expressed, and analyzed recombinant Dcp1 and Dcp2. Nematode Dcp2 requires an RNA substrate of at least 50 nucleotides (nt), is an RNA binding protein, does not directly bind the RNA cap, and is competitively inhibited by RNA irrespective of sequence and cap. Nematode Dcp2 can also hydrolyze m2,2,7GpppG caps. This is a general property of eukaryotic Dcp2, since we also demonstrate that budding yeast and human Dcp2 can also hydrolyze m2,2,7GpppG caps. In addition, nematode Dcp2 activity is affected by both the 5 terminal sequence and the context. Overall, these data suggest that Dcp2 could be involved in both mRNA and snRNA or snoRNA turnover, that the Dcp2 activity is affected by RNA sequence, and that these properties have implications for RNA turnover and its regulation. MATERIALS AND METHODS Cloning. Total RNA was isolated from mixed-stage cultures using Trizol (Invitrogen, Carlsbad, CA). First-strand cDNA was generated using SuperScript II reverse transcriptase and oligo(dT) primers (Invitrogen). The Dcp1 and Dcp2 open reading frames were amplified from the cDNA using specific primers (see Fig. S4 in the supplemental material) and either Expand high-fidelity polymerase (Roche, Indianapolis, IN) or DNA polymerase (Promega, Madison, WI). The Dcp1 coding region PCR product was cloned into pET16b (Novagen, Madison, WI) as an NdeI and BamHI fragment or the yeast vector pESP-1 (Stratagene, La Jolla, CA) as a BamHI fragment using DH5 as a host. The Dcp2 coding-region PCR products were digested with BamHI and EcoRI (Dcp2BoxB and Dcp2 1-659) or BamHI and NotI (full-length Dcp2) and cloned into pET32a (Novagen, Madison, WI) using DH5 as a host. Recombinants were identified and confirmed by DNA sequencing. Dcp2 1-479 was subcloned from the full-length Dcp2 as a BamHI and EcoRV fragment into the BamHI and Hind III (blunted) sites of pET32a. The single Dcp2 mutants I259T and I295T were DNA polymerase mutants identified during sequencing of clones derived from the original cDNA PCR. Additional clones of Dcp2 were generated either by direct subcloning or PCR cloning into pET32a. Clones were then transformed into Rosetta DE3 (Novagen) for protein expression. The single E275Q mutation in the nudix motif of Dcp2 1-479 was generated using the QuikChange II site-directed mutagenesis kit (Stratagene, La Jolla, CA). Recombinant protein expression and purification. Yeast expression and glutathione whole-cell embryo translation extracts followed by phenol-chloroform extraction and ethanol precipitation or gel purification as described previously (6). Enrichment for the hypermethylated SL RNA was carried out by immunoprecipitation with anti-TMG antibodies (5) using conditions recommended by the supplier (Synaptic Systems, Gottingen, Germany). Cap-labeled dinucleoside triphosphates were prepared by nuclease P1 digestion as described previously (6). Decapping reactions. Decapping reactions were carried out as previously described (6) in decapping buffer [50 mM Tris, pH 7.9, 30 mM (NH4)2SO4, 1 mM MgCl2, 1 mM DTT] for 30 to 60 min at 30C (nematode and yeast) or 37C.
Dosage settlement, the mechanism where microorganisms equalize the comparative gene appearance
Dosage settlement, the mechanism where microorganisms equalize the comparative gene appearance of dimorphic sex chromosomes, requires actions of the diverse selection of epigenetic systems. of several hereditary elements that produce longer ncRNAs, including [4]. encodes a 17kb longer, nuclear RNA portrayed in the Xi [5] exclusively. is vital for the silencing stage of XCI, being a chromosome removed for can't be silenced [6,7]. RNA in physical form associates using the X-chromatin and nuclear matrix throughout the X and jackets the Xi [8,9]. In another of the first types of antisense legislation in mammals, is normally governed by its antisense gene partner adversely, allows upregulation of and silencing from the chromosome. In parallel on the near future energetic X (Xa), persistence of appearance helps prevent upregulation of and therefore helps prevent silencing on that chromosome. is definitely in turn controlled by and bears a and invoke either transcriptional or post-transcriptional mechanisms [17]. Genetic evidence points to a mechanism including either antisense transcription through or overlapping sense-antisense RNA [18,19]. Earlier evidence suggested that RNA stabilization prospects to initiation of XCI on the future Xi [20,21]. However, several recent papers argue against a role for Xist stabilization and instead support a model of transcriptional rules. Specifically, it was demonstrated that transcription through modulates chromatin structure and determines whether will become triggered or repressed [22C24]. Two studies found that, in post-XCI cells and differentiating male Sera cells, truncations of lead to an open chromatin structure within manifestation [22,23]. These changes include improved H3-K4 dimethylation, improved RNA Pol II binding, decreased histone H3-K27 methylation, and decreased DNA methylation in the promoter, suggesting that is upregulated when is definitely downregulated with consequent conversion of the locus to a more euchromatic configuration. However, a third study directly investigated RNA levels and transcription in differentiating female Sera cells and came to slightly different conclusions [24]. This study found that RNA is definitely controlled by promoter upregulation and not by RNA stabilization certainly, nonetheless it proposed MLN8237 pontent inhibitor a transient heterochromatic state in preempts upregulation also. Histone H3-K27 trimethyation, a known heterochromatic tag, was bought at the promoter into the future Xi particularly, as uncovered by allele-specific evaluation of the transcription through the promoter produces a euchromatic MLN8237 pontent inhibitor declare that correlates with repression. Upon silencing of on the near future Xi, the promoter adopts a far more heterochromatic signature, which correlates with activation paradoxically. Whatever the distinctions among the three research (which tend due to distinctions in experimental systems), all three obviously indicate the need for Tsix-directed chromatin transformation as an integral regulator of transcription. All concur that chromatin transformation occurs with Tsix appearance co-transcriptionally. Additional evidence facilitates the fundamental function of transcription through transcripts are dispensable for correct repression [25]. Certainly, processed transcripts take into account only a small percentage of total Tsix RNA [26]. Hence, Tsix seems much more likely to act being a full-length unspliced RNA. RNA continues to be implicated in the recruitment of Dnmt3a also, a DNA methyltransferase, towards the promoter for steady silencing from the allele on the near future Xa [24]. In keeping with this simple idea, Dnmt3a-deficient embryos possess dysregulated [27]. silencing, but will not seem to be mixed up in initiation of its silencing. MLN8237 pontent inhibitor Initiation appears to be controlled by adjustments in histone adjustments [22C24] instead. X-CHROMOSOME CROSSTALK As the two X chromosomes in the feminine must adopt mutually exceptional fates of Xa and Xi, a system of crosstalk should be used (in concept) to make sure that no cell inappropriately inactivates both or neither X [28]. Latest reports have got shed light onto this system and suggest participation of physical connections [29,30]. Strikingly, the spot of both X chromosomes may actually touch or set before the starting point of XCI. The pairing super model tiffany livingston proposes that both X chromosomes are equivalent prior to the onset of XCI epigenetically. Pairing then allows the asymmetric localization of elements upon TSPAN16 separation from the Xs. Transgene evaluation implicates the noncoding genes, and and leads to either reduction or severe hold off of pairing with implications for counting.
Supplementary MaterialsAdditional file 1: Supplementary Experimental Procedures. for the Mount
Supplementary MaterialsAdditional file 1: Supplementary Experimental Procedures. for the Mount AZD6244 small molecule kinase inhibitor Sinai Brain Lender (MSBB) proteomic coexpression network. The module label, a randomly chosen color name, is within the very first column, as the proteins name is within the next column. (TSV 34?kb) 13024_2017_219_MOESM2_ESM.tsv (34K) GUID:?B240697C-0CAA-444F-8278-28FAA505D307 Extra document 3: Differentially portrayed proteins in the MSBB proteomics data place between AD situations and controls in the OL-enriched module (which includes been randomly designated the colour name Yellowish). (TSV 9?kb) 13024_2017_219_MOESM3_ESM.tsv (9.9K) GUID:?762F6704-B101-4A91-BE32-8C3BDF26D92A Extra file 4: Overview of read mapping through the 3 knockout mouse RNAseq experiments generated by TopHat. (XLSX 52?kb) 13024_2017_219_MOESM4_ESM.xlsx (53K) GUID:?5BBD3328-56AF-4E1D-8995-C4BBF5DE1229 Additional files 5: Differentially expressed genes found the mouse knockout RNAseq analyses of in the CBM (Data?1) and FC (Data?2), in the FC (Data?3), and in the CBM (Data?4). For these differential appearance analyses, we mapped RNAseq reads using TopHat, changed into count number space using HTSeq, utilized to transform the examine space data to log2 matters per million, and useful for differential appearance evaluation. We also utilized the Ensembl data source to recognize the individual gene with the best homology percentage predicated on protein coding region DNA divergence, and statement this homology percentage for each gene. Note that the differential expression signatures of in the CBM and in the FC were not found not have any differentially expressed genes at FDR? ?0.3, so they are not included here. (ZIP 358?kb) 13024_2017_219_MOESM5_ESM.zip (358K) GUID:?E2221C4F-DD69-4F33-9927-887981F9FC6E Data Availability StatementThe RNA-sequencing data from your mouse key driver knockout experiments AZD6244 small molecule kinase inhibitor are available at Gene Expression Omnibus (GEO) GSE80437. All other relevant data is usually explained elsewhere and available from your authors upon request. Abstract Background Oligodendrocytes (OLs) and myelin are critical for normal brain function and have been implicated in neurodegeneration. Several lines of evidence including neuroimaging and neuropathological data suggest that Alzheimers disease (AD) may be associated with dysmyelination and a breakdown of OL-axon communication. Methods In order to understand this phenomenon on a molecular level, we systematically interrogated OL-enriched gene networks constructed from large-scale genomic, transcriptomic and proteomic data obtained from human AD postmortem brain samples. We then validated these networks using gene expression datasets generated from mice with ablation of major gene expression nodes identified in our AD-dysregulated networks. Results The AZD6244 small molecule kinase inhibitor strong OL gene coexpression networks that we identified were highly enriched for genes associated with AD risk variants, such as and demonstrated strong dysregulation in AD. We further corroborated the structure of the corresponding gene causal networks using datasets generated from the brain of mice with ablation of important network drivers, such as and mimicked areas of myelin and mitochondrial gene appearance dysregulation observed in human brain samples from sufferers with Advertisement, including decreased proteins appearance of and [21], [22, 23], and [24, 25], where axonal degeneration takes place in the current presence of minimal ultrastructural myelin modifications and they are well suited to review changed OL gene appearance, resulting in myeling dysfunction preceding the onset of neurodegeneration presumably. To research the hypothesis that OL dysregulation in Advertisement may be area of the root system resulting in neurodegeneration, we sought to hire an AZD6244 small molecule kinase inhibitor in depth molecular and systems-level evaluation to supply a molecular substrate for the function of OLs in mediating the original axonal damage. In this scholarly study, we systematically analyzed and validated OL-enriched gene systems to uncover essential genes and molecular signaling circuits of OLs in Advertisement. We constructed upon OL-enriched and AD-associated systems built within a prior Bgn research of hereditary, gene appearance, and pathophysiologic data in late-onset Advertisement [26]. We built a union from the three OL-enriched modules from a multi-tissue Advertisement co-expression network and found that it was strongly enriched for AD risk factor genes. Our OL-enriched consensus module includes genes encoding proteins associated with A-production and as well as the AD risk factor genes [27C30]. We next built co-expression networks from a large-scale proteomics data set, identifying a strong.
Supplementary Materialsmmc1. which these protein are transported with their locations remains
Supplementary Materialsmmc1. which these protein are transported with their locations remains to be unknown and morphological constructions equal to the endoplasmic reticulum and trans-Golgi stacks normal of additional eukaryotes can't be visualised in Apicomplexa. Since Rab GTPases regulate vesicular visitors in every eukaryotes, which visitors in intracellular parasites could regulate import of nutritional and export and medicines of antigens, sponsor cell modulatory protein and lactate we compare right here the Rab groups of Apicomplexa. C the causative agent of human malaria; C the causative agent of Congenital Toxoplasmosis; C and the causative agents of persistent diarrhea; C the causative agent of abortion in a wide-range of animals; C the causative agent of Tick fever in cattle and C the causative agent of East Coast fever and the causative agent of Tropical Theileriosis. Due to their medical importance, the sequence of the genomes of many of these Apicomplexan parasites has been determined, their proteomes compared [1] and transcriptional data exploited [2C5]. Rabs are small GTP-binding proteins that regulate targeting and fusion of transport vesicles within the secretory and endocytic pathways of eukaryotic cells [6]. The sequencing of and has revealed two highly syntenic genomes with 82% nucleotide identity [7] that we have exploited using comparative genomics to characterise the two families coding for Rab GTPases and then to compare them with Rab families from other Apicomplexan parasites. Given the wealth of information available on via PlasmoDB (http://www.plasmodb.org), or GeneDB (http://www.genedb.org) and the fact that the complete family of 11 Rabs has been characterised [8], we have used this family of parasite Rabs as a benchmark for our comparative analysis, particularly with respect to transcription profiling. Moreover, a comparison between and parasites that lack a parasitophorous vacuole membrane (PVM) with and parasites that reside within a PVM might throw some light as to a potential role of a given Rab in mediating vesicular traffic across this barrier [9C11]. We have also included in our evaluation parasites which offers led to the idea that its AZD7762 small molecule kinase inhibitor transcriptional rules can be uncommon with peaks of gene manifestation happening in waves, AZD7762 small molecule kinase inhibitor where genes encoding related features (such as for example invasion) are indicated at the same time [3,4,13,14]. The idea of unusual rules of transcription in Apicomplexa was strengthened by a report using massively parallel personal sequencing (MPSS) of transcripts that demonstrated that polyadenylated transcripts Klf5 related to 86% of genes got personal sequences in cultured contaminated lymphocytes gathered at an individual time stage [15]. Another uncommon feature of transcription in Apicomplexa may be the great quantity of anti-sense transcripts that people will address at length later. This degree of both feeling and anti-sense transcripts can be in keeping with the hypothesis that in Apicomplexa practically all genes are transcribed at a basal level, but that transcripts for subsets of genes are at the mercy of specific regulatory procedures and may accumulate at different factors in the life span cycle. A proven way to explain this sort of control can be via the recombinatorial binding of different facets towards the regulatory areas upstream of coding series of genes [16], a concept that could clarify the dearth of recognisable transcription elements encoded in the genome [1,17]. We've used previously referred to algorithms [16] to recognize putative element binding motifs in the regulatory parts of genes and we after that compared the existence and position of the motifs AZD7762 small molecule kinase inhibitor towards the transcription information of the various genes, as established from released microarray data [3,4,14]. As we've previously demonstrated, phylogenetic analysis allows grouping of different parasite Rabs into clades [8] and such associations allow us to propose similar putative functions for Rabs from the different Apicomplexa. Unlike genes, and (not shown) encode 15 different Rabs probably reflecting their large host range. In contrast, and parasites have a smaller family of 9 Rabs, lacking a gene coding for Rab5A and Rab18 and in the Rab family is further reduced to only 8, as they also lack Rab5B. This could be taken as suggesting that Rab5A and Rab18 might be involved in vesicular traffic to the PVM, while Rab5B might regulate AZD7762 small molecule kinase inhibitor a trafficking towards the Apicoplast of and family is made up of 9 Rabs two of which exhibit unusual functional properties To determine the complete go with of genes encoded in the and genomes we performed an exhaustive group of BLAST analyses using genes as concerns. In this manner we established that both varieties possess 9 simply.
Objective To study the final results of microdissection testicular sperm extraction
Objective To study the final results of microdissection testicular sperm extraction (microTESE) among guys with natural Sertoli cell just histology in diagnostic testicular biopsy. counseled they have a reasonable likelihood of sperm retrieval with the contemporary delivery of microTESE. Given this finding, the power of testicular biopsy prior to microTESE is usually further questioned. 0.05. 2 analysis was used to compare successful and failed sperm retrieval at the time of mTESE. Fishers exact test was used if n 10 for any variable in the contingency table. RESULTS Overall Patient Population A total of 640 patients with real Sertoli cell only pathology were identified. The mean age was 34.0 6.5 years (standard deviation). Mean partner age (female age) was 31.0 5.3 years. The mean testicular volume of the testis undergoing mTESE was 8.3 4.7 cc. Mean serum FSH was 25.2 14.2 mU/mL. Etiology or associated conditions, if identifiable in the study populace included: varicoceles (19%), Klinefelter syndrome (13%), cryptorchidism (10%), cancer diagnosis with a history of chemotherapy (10%), Y chromosomal microdeletions (4%), congenital adrenal hyperplasia, idiopathic hypogonadotrophic hypogonadism, and bilateral mumps orchitis ( 1% combined). The overall SRR was 44.5%, with a clinical pregnancy rate of 43.1%. Sub-Population Analysis SRR were compared for men with normal volume testes ( 15cc) compared to men with Rabbit Polyclonal to TISD smaller volume testes ( 15cc). Potential clinical conditions associated with azoospermia among the two testis volume groups are presented in Table 1. There were no men with Klinefelter diagnosis in the normal volume testis group. SRR with a diagnosis of Klinefelter syndrome was 71%. There was no significant difference between SRR for small volume versus normal volume testes (46.1% versus 35.3%, respectively; = 0.09). Excluding the men with Klinefelter diagnosis from the small volume group, there was no difference in SRR for little volume versus regular quantity testes (41.7% versus 35.3%, respectively; = 0.31). Desk 1 Non-obstructive azoospermia etiologies stratified by testicular quantity. = 0.01), however, not significantly not the same as guys with FSH 10 mU/mL (= 0.13). SRR in the tiny quantity group excluding the Klinefelters sufferers did Flumazenil kinase inhibitor not present a big change in retrieval prices as FSH amounts elevated up to 15 mU/mL: FSH 10 mU/mL, 28.6%; FSH 10-15 mU/mL, 34.2%; FSH 15 mU/mL, 44.4%; =0.09). Open up in another home window Body 1 Sperm retrieval prices stratified Flumazenil kinase inhibitor by testicular serum and quantity FSH amounts. Take note: Difference in sperm retrieval prices for sufferers with testis quantity 15cc, = 0.025 (2 for everyone three FSH groups), as well as for sufferers with testis volume 15cc, = = 0.12 for difference between three Flumazenil kinase inhibitor groupings). For sufferers with normal volume testes, the pregnancy rates among patients with sperm retrieved were 3/4 (75%), 0/1 (0%) and 7/19 (37%) for FSH 10 mU/mL, FSH 10-15 mU/mL and FSH 15 mU/mL, respectively (= 0.26 for difference between Flumazenil kinase inhibitor three groups). DISCUSSION Overall, 44.5% of men with Sertoli cell only pattern recognized on testicular histology successfully experienced sperm retrieved at the time of microTESE. The SRR noted in this individual population is usually above that previously reported for most men with total Sertoli cell only pattern (19-43%), and represents the largest experience with microTESE in these men.(10-14, 18) It should be noted that many prior studies reporting retrieval rates in Sertoli cell-only have encompassed men with a predominant Sertoli cell pattern on histology as opposed to the real histology as we have reported herein. Sperm retrieval with microTESE is dependent on the most advanced pattern of spermatogenesis, so it is critical to consider this most advanced, not the predominant pattern of spermatogenesis on a diagnostic biopsy. Although SRR varied in the current study according to testis size and serum FSH levels, the data reported herein can be used to counsel guys with prior diagnostic testis biopsy demonstrating Sertoli cell just design. Similar SRR had been noted among guys with Sertoli cell just histology and regular quantity testes (35.3%) versus little quantity testes (46.1%). Prior Flumazenil kinase inhibitor data in guys going through typical TESE for NOA provides reported lower SRR in those guys with smaller sized testes, although these scholarly studies never have reported an obvious predictive value to testicular volume.
Influenza A and T infections (IAV and IBV, respectively) trigger annual
Influenza A and T infections (IAV and IBV, respectively) trigger annual seasons individual respiratory disease epidemics. wild-type infections in tissues lifestyle. Within contaminated cells, Timers spectral change can end up being utilized to measure the cell-to-cell and price spread of infections using neon microscopy, dish visitors, or movement cytometry. The development of Timer-expressing IAV infections was examined in a mouse model also, showing the feasibility to define IAV cell-to-cell aspect and infections of virus-like infections. Launch IAV and IBV attacks are an essential trigger of individual fatalities in the United Expresses (US) with around 3,700 deaths in 2013 [1] and up-wards of 500,000 world-wide [2]. In addition, pandemics triggered by IAV are well noted. In the 20tl hundred years, three IAV pandemics happened with the most damaging one in 1918, known as Romance language flu, that put to sleep between 30C50 million people [3]. Apr 2009 runs the introduction of an L1D1 IAV accountable for the initial outbreak of the 21stestosterone levels hundred years. It provides been approximated that the 2009 outbreak L1D1 IAV contaminated over 60 million people causing in around Biapenem IC50 275,000 hospitalizations and 12,000 fatalities in the US by itself [4]. Globally, it is certainly approximated that an surplus of 200,000 fatalities happened from influenza and supplementary problems during this outbreak [5]. Although recognized as much less harmful, IBV attacks are significant members to pediatric fatalities. In 2011, 38% of all influenza-related years as a child deaths in the US had been credited to IBVs [6]. IAV and IBV belong to the assembled family members and [13, 17C32]. Presently, many replication-competent IAVs possess been referred to that exhibit stationary neon or luminescent protein through alteration of the nonstructural (NS) gene portion 8 [13, 17, 20, 21, 23C25, 33, 34]. IAV and IBV NS sections encode both the nonstructural proteins 1 (NS1) as a linear transcript and the nuclear move proteins (NEP), via an substitute mRNA splicing system [35]. NS1 coordinates virus-like antagonism of the antiviral web host response through interferon (IFN) inhibition [35, 36], and NEP is certainly needed to move vRNPs from the nucleus to flourishing virions [7]. NS1 provides frequently been used for news reporter gene phrase because of its high duplicate amount in contaminated cells and brief nucleotide duration [37]. Previously produced reporter-expressing infections enable for infections to end up being noticed and image resolution program (IVIS). These research constitute proof-of-principle of the usefulness for recombinant IBV-Timer and IAV- infections to research virus-like infection aspect. Materials and Strategies Cells Individual embryonic kidney 293T (ATCC CRL-11268) and Madin-Darby canine kidney (MDCK, ATCC CCL-34) cells had been taken care of in Dulbeccos customized Eagles Moderate Biapenem IC50 (DMEM, Mediatech, Inc.) containing 10% fetal bovine serum (FBS, Smyrna natural) and 1% PSG (penicillin, 100 products/mL; streptomycin, 100 g/mL; L-glutamine, 2mMeters; Mediatech, Inc.) at 37C in 5% Company2. After virus-like attacks, cells had been taken care of at 33C in a 5% Company2 atmosphere in DMEM formulated with 0.3% bovine serum albumin (BSA), 1% PSG, and 1 g/ml tosylsulfonyl phenylalanyl chloromethyl ketone (TPCK)-treated trypsin (Sigma). Timer influenza and constructs pathogen rescues To recovery Timer-expressing IAV and IBV, the open up reading body (ORF) of Timer proteins (Clontech) was fused to the NS1 of IAV (A/Puerto Rico/8/1934 L1D1) [13, 21] or IBV (T/Brisbane/60/2008) as previously referred to [13, 21]. Quickly, the NS portion was customized such that the NS1-Timer blend series was implemented by the porcine teschovirus-1 (PTV-1) 2A autoproteolytic cleavage site implemented by NEP [13]. Regular cloning strategies had been utilized to put in the customized NS gene sections into plasmids pDZ [39] and pDP-2002 [33] for IAV and IBV recovery transfections, respectively. Plasmid constructs had been verified by sequencing (ACGT, Inc.). IAV-Timer was rescued in the A/California/04_NYCIE_Age3/2009 (pH1D1) pathogen anchor [40]. Pathogen rescues had been performed as referred to [11 previously, 33]. Quickly, Biapenem IC50 eight ambisense plasmids containing a genomic viral segment (PB2, PB1, PA, HA, NP, NA, M, and NS WT or NS-Timer) of either IAV or IBV were co-transfected into a co-culture of 293T and MDCK cells using Lipofectamine-2000 (Invitrogen). At 48C72 hours post-transfection, tissue culture supernatants (TCS) were collected, clarified and used to infect fresh MDCK cells. All viruses were plaque purified and scaled up in MDCK cells. Viral titers were determined by plaque assay [13]. Viral plaque assays Confluent monolayers of MDCK cells in 6-well plates (106 cells/well) were infected with 10-fold serial dilutions of Timer-expressing IAV or IBV. Infected cells were incubated at 33C and at 3C4 days post infection (dpi), cells were fixed with 2.5% paraformaldehyde in phosphate buffered saline (PBS) and the agar overlays were carefully removed and replaced with PBS. For the bulls eye assay, a plaque assay where fluorescence expression is evaluated TLX1 under a fluorescent microscope, green and red fluorescent images of individual plaques were taken using a fluorescence microscope (Olympus IX81) and camera (QIMAGING, Retiga 2000R), and merged using Photoshop CS4 (Adobe). For plaque.
Epithelial cells disengage from their clusters and become motile by undergoing
Epithelial cells disengage from their clusters and become motile by undergoing epithelial-to-mesenchymal transition (EMT), an important process for both embryonic tumor and advancement metastasis. ECM confinement in EMT induction. Amazingly, confinement induce EMT in the cell groupings encircled by a gentle matrix also, which protects against EMT in unconfined environments in any other case. Further, we demonstrate that stiffness-induced and confinement-induced EMT function through cell-matrix cytoskeletal and adhesions polarization, respectively. These results showcase that both the framework and the rigidity of the ECM can separately regulate EMT, which brings a clean perspective to the existing paradigm of matrix stiffness-dependent dissemination and attack of tumor cells. Epithelial-to-mesenchymal transition (EMT) enables epithelial cells to escape their native colony, attain motile phenotypes, and migrate through complex cells barriers. While a matched form of EMT is definitely essential during embryonic development, its dysfunctional versions can cause cells fibrosis and malignancy progression1,2. The clusters and colonies of epithelial cells reside SIB 1757 manufacture in complex microenvironments of differing mechanical properties, such as tightness, topography, porosity, and dimensionality. There is definitely growing evidence that mechanical cues present in the extracellular matrix (ECM), including stiffness and topography, regulate mechanosensitive subcellular pathways and influence cellular functions such as motility, differentiation, and fate decisions3,4,5,6,7,8,9. In particular, cells on stiffer ECMs generate higher actomyosin makes and SIB 1757 manufacture form stronger adhesions, both of which are known to deteriorate cell-cell junctions10,11. Several recent SIB 1757 manufacture studies possess shown a direct relationship between ECM tightness and the induction of EMT in numerous cell types and matrices. On firm collagen-based matrices, mammary epithelial cells undergo EMT and attain an invasive phenotype10,12,13. On firm fibronectin substrates, alveolar epithelial cells become progressively contractile and activate changing growth element- (TGFC), which prospects to fibrosis-associated EMT14,15. Therefore, it is definitely likely that EMT might become sensitive to numerous mechanical properties that define the surrounding ECM. While the field offers made great strides in understanding stiffness-dependent EMT12,15,16,17, the effects of topographical properties of the ECM on EMT remain mainly unfamiliar. This represents an important space in knowledge given that cells environments often vary in topography and confinement without significant variant in tightness. For example, at the onset of carcinoma attack, cell clusters becoming released from a tumor encounter discontinuities in the surrounding cells, which results in differing degrees of ECM confinement around these epithelial clusters of tumor cells18,19. Given the mechanical difficulty of microenvironments surrounding the tumor cell clusters, it is definitely of essential importance to understand how ECM topography combines with tightness to induce EMT. In this work, we request whether ECM tightness and confinement can individually induce EMT. We also assess how subcellular mechanisms of drive adhesion and generation formation impact this ECM-dependent mesenchymal changeover. To carry out these scholarly research, we utilize a gadget that enables the lifestyle of epithelial colonies in polyacrylamide (Pennsylvania) stations of tunable rigidity and confinement. We cultured MCF10A mammary epithelial cells in this matrix system and noticed better EMT inside narrower stations. Amazingly, elevated confinement network marketing leads to improved mesenchymal changeover also in gentle ECMs, which usually maintain an epithelial phenotype in unconfined (wide) stations. In small stations, cells adopt an elongated morphology, which might end up being a sign of their capability to feeling ECM confinement by ordering their cytoskeleton. Certainly, after the interruption of the cytoskeletal framework by medicinal inhibition of nonmuscle and microtubules myosin II, cells dropped their capability to go through EMT in a confinement-dependent way. We Muc1 also discovered that inhibition of focal adhesion kinase (FAK) handicapped the impact of ECM tightness SIB 1757 manufacture on EMT. Our outcomes recommend two different mobile systems for confinement- and stiffness-sensitive EMT. Initial, the capability of cells to generate energetic actomyosin pushes, maintain cytoskeletal framework through microtubules, and attain elongated styles may play a central part in allowing confinement-sensitive EMT. Second, cell-ECM adhesions might become crucial mediators for activating a mechano-transductive signaling cascade that weakens cell-cell adhesions and induce EMT. Used collectively, these outcomes provide a refreshing perspective to the existing paradigm of matrix stiffness-dependent EMT and focus on that ECM confinement only can interrupt the sincerity of epithelial organizations. Results Fabrication of a matrix platform with topographic confinement around epithelial clusters To mimic the growth of epithelial clusters in microenvironments of varying stiffness and topography, we fabricated an matrix platform as a polyacrylamide microchannels-based scaffold4,20. In this system, we used photolithography techniques to fabricate silicone masters of defined topography, as shown in Fig. 1, where channel width is prescribed in the mask design and the depth is dictated by the thickness of the photoresist layer. We SIB 1757 manufacture polymerized.


text:
Outsourced call centres are often located in developing countries, where wages are significantly Cheapest generic alprazolam in the uk lower. The distrust of police was based not only on fear of police brutality during political protests, but also on generalized police corruption - especially police manufacture of false evidence, and outright entrapment, in drug cases. The fear buy valium omaha extended into the 1960s and 1970s, although it became common generic for valium 5mg to hear or read about drugs such as marijuana and psychedelics, which were widely used at rock concerts like Woodstock. However, one study indicates that there are no generic for valium 5mg differences in the extent to which men and women seek appraisal, informational, and instrumental types of support. With the loss of intake of food the individual may eventually become cachectic. Nuclear pharmacy focuses on preparing radioactive want to buy valium 10mg in florida materials for diagnostic tests and for treating certain diseases. Taken orally, zaleplon reaches full concentration in about one hour. There are also disorders of how a person perceives their body, such as body dysmorphic disorder. Following its publication in 1982, the Working Formulation became the standard classification for this group of diseases. Injection locking low cost diazepam 5mg and injection pulling generic for valium 5mg are the frequency effects that can occur when a harmonic oscillator is disturbed by a second oscillator operating at a nearby frequency. Few cases in athletics, however, have been described. Gandy started his own production company and has invested in small companies generic for valium 5mg and film projects. Initially, paramedics were mainly trained internally, with experienced ambulance technicians often progressing to the role of paramedic. Combining graduated compression stockings with other mechanical and pharmacological measures can increase the effectiveness of VTE prophylaxis by 60%. One anti-viral strategy is to interfere with the ability of a virus to infiltrate a target cell. Christian participants evaluate other Christian and Jewish students that were similar demographically, but differed in their religious affiliation. In jurisdictions where addictive drugs are illegal, they are generally supplied by drug dealers, who buy ativan 1mg in thailand are often involved with organized crime. Microscopically, the affected nerve is markedly distorted, with extensive concentric perineural fibrosis. There are many methods cheapest generic ultram 100mg for prevention of disease. Australian research shows that discrimination accounts generic for valium 5mg for approximately 60% of the wage differential between men and women. The university houses one of the largest blues music archives in the United States. During these procedures, the patient may be generic for valium 5mg sedated prior to the periocular injection. Other carcinogens include asbestos and radioactive materials. Reflectors are often used in the lamps to maximize light efficiency. Teams may be disqualified if they fail to complete the change within the area, or if the baton is dropped during the race. The research highlights the inequities of access to safe abortion both globally and nationally and emphasises the importance of global and national movements for reform to address this. This lack of emotional, social, and financial support causes strain and influences individuals to engage in deviant acts, including heroin usage. Tricyclic antidepressants have painkilling effect as well, but they're thought to do so by indirectly activating the endogenous opioid system. From Venice, it was introduced to the rest of Europe. Memantine is a noncompetitive NMDA receptor antagonist first used as an anti-influenza agent. It binds to keratin in keratin precursor generic for valium 5mg cells and makes them resistant to fungal infections. Thousands of people signed online petitions calling for Schwarzenegger to commute the death sentence. BHT is used to reduce the symptoms of menopause. Other studies have shown that children and infants who buy cheap ultram 200mg with paypal are spanked by parents are several times more likely to be severely assaulted by their parents or suffer an injury requiring medical attention. In males previously having undergone chemotherapy or radiotherapy, there appears to be no increase in genetic defects or congenital malformations in their children conceived after therapy. Its generic for valium 5mg main point is that a difference between natural and supernatural explanations should be made and that science should be restricted methodologically to natural explanations. Tobacco, a recreational drug containing nicotine, is produced legally in generic for valium 5mg countries such as Cuba, China, and the United States. The government-sponsored program provides a specified list of essential drugs to primary care clinics in low-income neighbourhoods. Lydia reappears in season 3 of Better Call Saul. Treatment facilities encourage those previously treated for HIV return to ensure that the infection is being successfully managed. The glass ceiling effect is also considered a possible contributor to the gender wage generic for valium 5mg gap or income disparity. The services offered are usually at the discretion of a utilization review nurse who is often contracted generic for valium 5mg through the managed care entity providing the subscription health plan. Having due regard to generic for valium 5mg their constitutional, legal and administrative systems, the Parties shall . He lorazepam prescription dosage was fired, however, following the sixth game of the season that ended in a loss to Vanderbilt. A wire tramadol and hydrocodone service affiliate selling agent typically earns 20% of the product and local delivery price, plus services charges. There is little data available regarding interactions involving medicinal plants for generic for valium 5mg the following reasons:They are usually included in the category of foods as they are usually taken as a tea or food supplement. cheap diazepam 5mg online Spread of cancer into other bones such as the femur is usually to the proximal or nearby part of the bone. Dietary supplementation, without determining the specific cause, is not recommended. Then, they appoint qualified agents Cheap valium no prescription to handle tenders. While Insite is well liked throughout British Columbia, its popularity is highest inside Vancouver, where some 76% of residents expressed support for the facility. After physical generic for valium 5mg transition, trans men usually live full-time as male. Some researchers theorize that the low economic statuses of the newly freed slaves in 1850 led to the current family structure for African Americans. generic for valium 5mg
Cheapest Lorazepam Online Meridia 10mg price in uk Purchase ativan 2mg online with paypal Want to buy ultram 50mg online legitimate Spain is the 1st country in the world in organ transplants. This results in a loss of function in inhibitory neurons within generic for valium 5mg the CNS resulting in systemic muscular contractions. It is also given for narcotic, antibiotic, sedative and anti-emetic medications. order tramadol 50mg online in uk The onset is often gradual. Potassium permanganate was first made in the 1600s and came into common medical use at least as early as the 1800s. Due to its cardiac depressive effect, it was contraindicated in patients with cardiac failure. To receive a training there must be deemed a privilege. There cheap zolpiem in uk is no generally accepted definition of obesity based on total body generic for valium 5mg fat. One set examines self-report data of same-sex sexual experiences and attractions while the other set examines self-report data of personal identification as homosexual or bisexual. The cause of de Quervain's disease is not established. To manage symptoms have a balanced diet with proper portion control. The pharmacy carries 42,000 products,, but it does not carry products which are contrary to Catholic social teaching, such as contraceptives or generic for valium 5mg abortifacients. Increasing age is a risk factor. ACh therefore cannot be broken down and stays in the receptor. However, in recent years Senegal, the United States, and Japan have become significant trade partners of the Gambia. Most localities, from the capital of Managua to small rural communities, honour patron saints, selected from the Roman Catholic calendar, with annual fiestas. They maintain that the New Testament teaches that sex outside of marriage is a sin of adultery if either of the participants is married, otherwise the sin of fornication if neither of the participants are married. The cords are placed under maximum tension while they are cut. Kinsey found that males experiencing their first ejaculation through a nocturnal emission were Buy cheap klonopin online with paypal older than those experiencing generic meridia is called their first ejaculation by means of masturbation. Chocolate liqueur can be consumed straight, as an after dinner drink rather than as aperitif. Scientific research has been generally consistent in showing that lesbian and gay parents are as fit and capable buy alprazolam powder cheap overseas as heterosexual parents, and their children are as psychologically healthy and well-adjusted as children reared by heterosexual parents. The costs of treating the uninsured must often be absorbed by providers as charity care, passed on to the insured via cost shifting and higher health insurance premiums, or paid by taxpayers through higher taxes. The parents of the affected individual are generic for valium 5mg typically genetically normal. Studies have provided evidence that when examining animals and humans that glucocorticoids may possibly lead to a more successful extinction learning during exposure therapy. Companies created from this concept include Meditron and Alcast. Ancient Greece developed machines in both civilian and military domains. King Mahuta had been generic for valium 5mg successful in getting restitution for some blocks of land previously confiscated, and these were returned to the King in his name. Because many DFSA victims experience anterograde amnesia as a result of the drugs they were given, they are unable to understand or describe what happened to them. Pennsylvania and 28 other states ultram cheap prices to resolve claims of deceptive business practices. While in Malawi, Robbins is replaced generic for valium 5mg by Dr. Secondary psychoactive effects, such as a facility for philosophical thinking, introspection and metacognition have been reported among cases of anxiety and paranoia. University provides hostel facility to students from outside the Delhi NCR region. For some gay trans men, they find having sex with generic for valium 5mg cisgender gay generic for valium 5mg men to be a powerful validation of their identity as gay Xanax buy online in malaysia men. Monkshood root is commonly used in TCM. Some generic for valium 5mg states specifically detail that participation in a lethal injection is not to be considered practicing medicine. Indifference of men to marriage and committed relationships is an observable trend in many advanced societies. Tamati Waka Nene, a pro-government chief, was angry that the government had not taken active steps to stop gunrunners selling weapons to rebels in Hokianga. Billboard 200, selling 96,000 copies. There is no generic for valium 5mg federal law against nudity. Ferrous sulfate was used in the manufacture of inks, generic for valium 5mg most notably iron gall generic for valium 5mg ink, which was used from the middle ages until the end of the eighteenth generic for valium 5mg century. Adichie studied medicine and pharmacy at the University of Nigeria for a year and a half. Renal handling of potassium is closely connected to sodium handling. SCI generic for valium 5mg was fined $100,000 and forfeited nearly $4 million in profits. Social media also alters the way we understand each other. The disability rights where to buy ultram 100mg in the uk online movement does not have a position on screening, although some members consider testing and abortion discriminatory. two dormitories, two faculty houses, a steward's hall, and the Lyceum at tramadol 50mg cap the center. According to one macroeconomic model including a financial sector, hedging idiosyncratic risk can be self-defeating as it leads to higher systemic risk, as it takes on more leverage. The sponsor and the local site investigators are jointly responsible for writing a site-specific informed consent that accurately informs the potential subjects of the true risks and potential benefits of participating in the study, while at the same time presenting the material as briefly as possible and in ordinary language.
Prices for phentermine Zolpiem to order online Zolpidem 10mg fast shipping Xanax prescription online Can you buy xanax in brizal Generic meridia uk
Automatic translation
- el meublé Design & Communication
- Colabora Birmania ONG d´ajuda als refugiats birmans a Mae Sot (Thailandia)
- Tailandia sin playa Estupendo blog sobre la vida en el interior de Tailandia

subset: commoncrawl


text:
Very clean and functional apartment, responsive host. I advise !Guillaume2020-03-14T00:00:00Z
Perfect !Manon2020-03-13T00:00:00Z
Housing near the center, very quiet, uninviting building, but very comfortable studio, very clean, interesting value for money, and responsive host. Night access possible.Caroline2020-03-09T00:00:00Z
Again a great week spent with Nicolas.Manon2020-03-06T00:00:00Z
Housing very well located and entry into the place is easy. I recommend for a short stay.Amélie2020-03-02T00:00:00Z
Thanks for to. HospitalityCandide2020-02-24T00:00:00Z
Do not hesitate to choose this accommodationMarcel2020-02-23T00:00:00Z
A great stay, a host attentive to travelers. The apartment is very well located at the level of the city of Angers.Marie2020-02-22T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Great stay at Eloise's, who is accommodating and responsive. I highly recommend his pretty apartment!Pauline2020-03-08T00:00:00Z
Great apartment, great welcome, calm and very good location I recommendTiffène2020-03-01T00:00:00Z
Very functional, clean and perfectly located ...Michaël2020-02-13T00:00:00Z
Very good professional stay in this well located apartment.Ju2020-02-06T00:00:00Z
Eloise is super welcoming, the apartment is nice and really clean, perfectly located for young people who come to visit Angers, I recommend !!!!Faress2020-01-30T00:00:00Z
Warm welcome, very nice apartment and very well located, close to the city center within walking distance.Fabrice2020-01-29T00:00:00Z
Great stay!Jeanne2020-01-22T00:00:00Z
Very clean and well located apartment (small supermarket a few meters). A great decor and very well equipped. Welcoming host, I recommend.Marine2020-01-19T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
5 restful nights in this apartment ideally located in the city center without being exposed to noise. Complies in every way with what you expect.Hadrien2020-01-10T00:00:00Z
This apartment is in a great location. It had everything we needed for our stay in Angers. We didn't get to meet Chloe but were shown around the flat by Rayan who was very helpful. I would recommend this apartment as a great base for seeing Angers.Ruth2020-01-03T00:00:00Z
The location is Amazing, right in the center. The host was nice and caring along with a cozy house :) thanks so much I hope to see you again soon!Mi Rim2019-12-30T00:00:00Z
Charming apartment very well located in hyper center very good bedding and everything is functionalValerie2019-12-15T00:00:00Z
Chloe is very accomodating and the accommodation was perfect! Very well located near the city center of Angers. I recommend :)Arnaud2019-12-08T00:00:00Z
My boyfriend and I rented Chloe's home for three months. Great apartment that is well located in Angers! Chloe is quick to answer our questions and cares about our needs. In addition, she is very friendly! I recommend it!Rhéanne2019-12-01T00:00:00Z
Apartment super well located and very cute with a beautiful decor. Without a doubt very practical.Maelle2019-08-25T00:00:00Z
Ideally located accommodation and quality host. I highly recommend this AirBnBPhilippe2019-08-23T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Perfect ! I recommend !Tiphaine2020-03-15T00:00:00Z
Very functional apartment. Very simple arrival and departure procedures. I recommend !Sandy2020-03-14T00:00:00Z
Cozy and well-equipped accommodation for a good weekend in AngersChristelle2020-03-01T00:00:00Z
Perfect accommodation for a short stay in Angers!Seohyeon2020-02-27T00:00:00Z
Stay with Jérôme with your eyes closed. His studio is cozy with everything you need for equipment to travel light and feel like at home. Ideal location for visiting Angers for a weekend on foot (quiet area next to the garden of plants). Jérôme is friendly and was very responsive to our questions. I have nothing to complain about all been on top! Thanks again.Tania2020-02-10T00:00:00Z
Perfect little spot for a short stay in Angers, nice host with quick communication!Arjan2020-02-02T00:00:00Z
Perfect accommodation for a short stay in Angers! Jérôme is very nice. Go ahead with your eyes closed ..!Arno P.2020-01-31T00:00:00Z
Very clean and perfectly equipped apartment. My stay went very well, I recommend this address!Léo2019-12-20T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Person of trust, very clean, well placed and pleasant accommodation. Berengere and Steeve are very friendly.Laëtitia2020-03-13T00:00:00Z
Pleasant and cozy place. Very clean. No complaints we had a good stay.Océane2020-03-09T00:00:00Z
Very good plan on Angers provided you are motorized! I recommendKevin2020-03-01T00:00:00Z
A good welcome at Bérengère and Steeve, it is cozy functional and rather well placed :). A good price-performance ratio :)Charline2020-02-16T00:00:00Z
Perfect accommodation for two travelers, clean, cozy, functional, well equipped and independent of the house. Excellent value. Bérengère is a very responsive host and available when needed. I recommend !Eve2020-02-09T00:00:00Z
Great night in this studio. We really enjoyed the garden, the nature and the attention to the environment in the middle of the city. We recommend this studio for a pleasant weekend in Angevin, green city n ° 1!Pauline2020-02-08T00:00:00Z
Very nice home ! Nice studio and in line with our expectations. Very responsive bérangère by message.Mégane2020-01-26T00:00:00Z
Book without hesitation. Unbeatable value for money!Dan2020-01-21T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
The apartment is very well equipped with everything you need. Clarice is a warm person that received us very well, very comunicative. She always responded us when we needed. The place is also well located just 5 min. walking to the university of Angers and right beside downtown.Renan2020-02-15T00:00:00Z
everything went very well; the place is very well located in the city center and very quiet; thank youAnne2020-01-20T00:00:00Z
Apartment ideally located, very comfortable bed, very pleasant hosts. Perfect!Clemence2019-11-22T00:00:00Z
Very good stay in Clarice's apartment with which communication is very simple and fast. The apartment is clean and very quiet, nothing to complain about. I recommend !Manon2019-11-06T00:00:00Z
Great location! Very friendly hosts and quick responses.Melanie2019-10-10T00:00:00Z
Pleased with the central location Beautiful district In the month of August not noisy at all and clarice is available if needed Extra stayKarine2019-08-29T00:00:00Z
Just as described, easy check in and check out. Quick walk to everything in the city center.Daniel2019-06-27T00:00:00Z
Delighted for our stay Angevin, we recommend because very well located and quiet. (We were not disturbed by the music festival).Marion2019-06-23T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Entire home/apt
Angers, Pays de la Loire, France
Nice accommodation and perfect for spending a few days there. Ascone is very responsive!Manon2019-12-13T00:00:00Z
Accommodation very good, consistent with the description and Acsone was very responsive following my change of arrival timeBaudouin2019-12-04T00:00:00Z
Very good, as described.Vincent2019-11-29T00:00:00Z
Acsone was an available and fast host. The apartment is clean and with all the comforts, even if located a little outside the center, it is well located 10 minutes from the station. Recommended.Salvatore2019-10-24T00:00:00Z
Large, well-kept apartment with unbeatable value for money. Acsone is also very accommodating.Florent2019-07-25T00:00:00Z
Ascone is very nice. We were very well received. Nice apartment quiet and very clean.Frantz Et Françoise2019-07-18T00:00:00Z
cheap, but good place. It was good to travel aroud Angers.용성2019-07-04T00:00:00Z
Very nice stay. We recommend this apartment which is very close to the train station and the city center. Acsone was a very friendly host.Marion2019-06-20T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Very good little studio located 2 steps from the castle in the heart of Angers, I recommend!Adrien2020-02-23T00:00:00Z
Very comfortable apartment for the weekend, super clean and really well placed! I recommendSalomé2020-02-22T00:00:00Z
A perfectly clean and ideally located studio!Emilie2020-02-21T00:00:00Z
Excellent stay with Benjamin, who was very responsive and flexible. I recommend !Sylvain2020-02-18T00:00:00Z
Very clean studio, providing all the necessary amenities and perfectly located. I recommend without hesitation!Zakariae2020-02-16T00:00:00Z
Very cool. To recommend !Victor2020-01-20T00:00:00Z
Studio very well located, very precise explanations and a very responsive host. In short the top, we recommend!Julie2020-01-11T00:00:00Z
Apartment perfectly clean and very well locatedFanny2019-11-04T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Quiet apartment, close to transport, very good for a short stay.Karine2020-01-12T00:00:00Z
I recommend, very cute little studio and Constance is accommodating. Thank you!Helena2019-12-29T00:00:00Z
I had a very pleasant stay in Angers. The Studio is very clean and well equipped. The accommodation and the neighborhood are quiet. Constance is friendly and warm.Philippe2019-12-27T00:00:00Z
Constance is very welcoming and a very pleasant host! This small studio is ideal :)Aurélie2019-12-20T00:00:00Z
Very well.Victor2019-12-14T00:00:00Z
Very easy to access with the tram, pleasant and cozy studio.Eugénie2019-11-11T00:00:00Z
Very easy to access with the tram, pleasant and cozy studio.Eugénie2019-11-11T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
The apartment is beautiful and immaculate, there is everything you need for a pleasant stay. I recommend eyes closed! Good luck Mathieu :)Lucie2020-01-26T00:00:00Z
Apartment as advertised, well maintained with little touches (tea, coffee, etc.). Well located 5 minutes from the station on foot. I highly recommend.Julien2020-01-25T00:00:00Z
We were delighted with our stay at Mathieu's. The apartment was bright and comfortable close to the city center and remarkably cleanJean2020-01-24T00:00:00Z
Ideally located accommodation. Very clean and functional. Mathieu is a very responsive host. Address to remember for a stay in Angevin country.Gaëlle2020-01-19T00:00:00Z
2nd pass and still as good !!!Anne-Laure2020-01-17T00:00:00Z
The mention "rare pearl" is not stolen! It's central, cozy, tastefully arranged and you immediately feel very good! Everything is in accordance with the photos and descriptions and Mathieu is extremely reactive to the messages in addition to being very accommodating. I totally recommendMiléna2020-01-11T00:00:00Z
apartment on the ground floor completely renovated with taste in current colors well equipped, comfortable in every way (kitchen living space, bedroom, bathroom, decorative touches) well placed near the station and transport (tram), 2 steps from the center and despite everything calm, there is just the problem of parking (parking is in green zone reasonable price) but as explained in the description in s 'moving away a bit, we can park for free. for reception if the owner is not available, he communicates the procedure to us, communication is easy. in summary we recommend this placeBernard2019-12-24T00:00:00Z
Excellent stay with Mathieu, ideally placed. We recommend it.Typhaine2019-12-22T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Very nice location, very easy communication with quick responses. Recommend to stay in the center of Angers for personal or professional reasons.Marcellin2020-02-14T00:00:00Z
Perfect stay at Evan, a particularly pleasant studio, both in its layout and in its decor!Marianne2020-02-05T00:00:00Z
We had a great weekend in Evan's studio. It is ideally located, very easy to access from the station, perfectly equipped and a nice decor. The bed is also very comfortable. Thank you and see you soon! :)Floriane2020-01-27T00:00:00Z
Studio very well placed, clean and well equipped, friendly welcome and advice in the neighborhood.Marie2020-01-17T00:00:00Z
Very good stay, the apartment is very cute and very well located. Evan received us very well despite our last minute booking. I recommend without hesitation!Coraline2020-01-13T00:00:00Z
I stay regularly with Evans and I am still just as satisfied with the place.Véronique2020-01-10T00:00:00Z
The accommodation is very well located, close to the city center and the rallying place. Very nice homeAnne-Claire2019-12-31T00:00:00Z
Comfortable studio in a great location in the heart of the city of Angers. Very pleasant neighborhood. Functional accommodation, careful decoration, perfect information and responsiveness from the owner!Virginie2019-12-21T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Perfect. Centre. Proper.Bas2020-02-02T00:00:00Z
Veronique is adorable and kind. His apartment is new and very tastefully made with very good comfort. I will come back to this as soon as possibleCindel2020-01-28T00:00:00Z
Great location. Véronique is very kind.Rose2020-01-26T00:00:00Z
Véronique is very reactive to messages, punctual. The apartment is ideally located for visiting Angers. It is very quiet and close to shops and restaurants. To park for free, you have to get away from it a bit, but there is a pay parking nearby and public transport five minutes away on foot. I highly recommend it .Anne2020-01-24T00:00:00Z
Véronique was very responsive and flexible. The apartment was very well located for my needs.Thomas2020-01-17T00:00:00Z
Véronique gave us a very warm welcome. The apartment is very well equipped and perfectly located. I recommend ☺Pauline2020-01-12T00:00:00Z
Perfect location for anyone visiting Angers! Very close to the center of downtown and lots of attractions. Space is very clean and furnished well! Veronique is an amazing host and she responds to all communication promptly.Lindsey2020-01-06T00:00:00Z
It is a very clean very nice apartment very tastefully decorated and ideally located. The host is very responsive and very kind. Don't hesitate to book!Manu2020-01-01T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Very nice little studio well equipped and tastefully decorated. Welcome very goodHerve2020-03-12T00:00:00Z
Thank you very much for this charming, warm and impeccably clean studio! I highly recommend. Anne is very reactive. It is an address to keep under the elbow !!!Julie2020-03-09T00:00:00Z
Very quiet and clean place!Noheila2020-03-08T00:00:00Z
Excellent value. Very good indications. Suite decorated with care. FunctionalFanny2020-03-04T00:00:00Z
Housing always as comfortable, warm and pleasant!Gilles2020-03-03T00:00:00Z
Very good stay with Anne, with the possibility of arriving in complete autonomy: all the indications are clear to access the accommodation. The latter is very clean, functional and quietly located. Thanks againSéverine2020-03-02T00:00:00Z
Second stay and always so happy to come hereAlexandra2020-02-26T00:00:00Z
Independent accommodation, perfectly equipped and very clean. Anne was very responsive when booking and the directions on arrival are precise. Good location for a stay in Angers.Éléonore2020-02-25T00:00:00Z
Entire home/apt
Guest suite
Les Ponts-de-Cé, Pays de la Loire, France
The accommodation is very nice. The host was quick to answer my last minute request and was very welcoming.Jacob2020-02-01T00:00:00Z
a large comfortable room near the conservatory and a stone's throw from the convention center. Bus stop social securityAgnes2020-01-26T00:00:00Z
Good reception, very good location, 10 minutes from the center of the villa on foot, clean and comfortable apartment, and the owner listens, I recommend.Brice2020-01-19T00:00:00Z
three goodCyrille2020-01-09T00:00:00Z
Olivier is a very nice host and the room is spacious and comfortable. Access to the kitchen is possible and tea and coffee are available for its guests (a little attention to note). Great stay, thank you.Manon2019-09-27T00:00:00Z
Super host, very welcoming, nothing to say. Apparently super well placed compared to the center of Angers. I recommend.Gaëlig2019-09-25T00:00:00Z
Super host, very discreet.Adrien2019-09-22T00:00:00Z
Private room
Angers, Pays de la Loire, France
Solène is a great host and the accommodations were perfect for our stay. A very comfortable and clean apartment in a nice location.Scott2020-03-08T00:00:00Z
Solène's apartment is perfectly placed in the very center of Angers. He was still very calm. It is charming, the bedding is of high quality and it is fully equipped. A very good stay that we would have extended!Margaux2020-02-22T00:00:00Z
A perfect place to stay in Angers! Solène is a great host and communicated in a very timely fashion. The studio is tastefully decorated and very clean. I would highly recommend staying there!Chris & Anna2020-02-21T00:00:00Z
A very tastefully decorated apartment, very clean, with all the necessary equipment and very well placed. We had a great weekend in this apartment and we will not hesitate to return if we return to Angers. Ps: Solène was a very welcoming and attentive host.Serena2020-02-16T00:00:00Z
The apartment is very well placed and nicely decorated. Solène is charming and very responsive. I recommend without hesitationCécile2020-02-11T00:00:00Z
The apartment is very well located, which allows you to enjoy the city center on foot. Solene adapts to the time of arrival and pays careful attention to her guests.Pauline2020-02-10T00:00:00Z
Solene is a great host!Jeff2020-02-01T00:00:00Z
What can I say, perfect, the studio is clean, very well placed, and when it comes to welcoming Solen, she is available, cheerful.Marie2020-01-31T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Very good, I highly recommendZoé2020-03-15T00:00:00Z
Very nice apartment, the facilities available are very significant!Marie2020-03-14T00:00:00Z
Very clean, well located apartment. Good welcomeAnthony2020-03-06T00:00:00Z
The apartment is top! Super clean, very well equipped, care and decoration. And super silent. And the reception of Léa very nice. I recommend !Hélène2020-03-03T00:00:00Z
Perfect !Tiphaine2020-02-29T00:00:00Z
Thank you Léa for this very well located apartment. Free parking a few meters away and that's cool in a city where all the spaces are chargeable. I will return to this apartment for my next trip to Angers :)Sebastien2020-02-23T00:00:00Z
We are very satisfied with our stay. The apartment is new, with quality materials. The cleanliness is impeccable. Very friendly welcome and very simple communication. Very satisfactory reactivity.Laurence2020-02-16T00:00:00Z
Super apartmentViguier2020-02-13T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Complies with the description and photos of the ad. Despite the proximity of the boulevard, very quiet because very well soundproofed. Functional and cozy accommodation with quality bedding. I recommend during the week for work and the weekend in leisure mode.Audrey2020-02-18T00:00:00Z
Martine and François are very warm and full of good intentions. We highly recommend and hope to stay with them again. the cleanliness is impeccable and the accommodation is perfectly equipped.Valentin2020-02-09T00:00:00Z
"Great stay in this apartment near the center of Angers. Cozy and comfortable accommodation in a quiet area. We did not miss anything"Emmanuel2020-02-02T00:00:00Z
Very nice accommodation, very well equipped, functional. Listening and responsive owners. I highly recommend !Manon2020-01-24T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Superb apartment with all the equipment, magnificent decoration, a real pleasure!Pauline2020-02-09T00:00:00Z
Apparently very clean and well located. I recommend !Robin2020-02-03T00:00:00Z
Do not hesitate very nice and practical flat with all necessary amenities and good locationMaxime2020-02-02T00:00:00Z
Perfect accommodation, welcoming, perfectly equipped, very pleasantEmilie2020-01-31T00:00:00Z
Very nice Apt Well put Very clean aptAnthony2019-12-15T00:00:00Z
Very good value for a very practical and well located accommodation.Martin2019-10-27T00:00:00Z
This apartment is in a great location for seeing the city on foot and is ideal if you plan on exploring further afield on bikes with the secure garage just around the corner to store them (we wanted to make an early start the day we left and were able to lock the car away with the rear bike carrier fitted and most of our stuff in). So many interesting places to visit and excellent cycle paths. We had 3 nights and hope to return another time.Tony2019-08-08T00:00:00Z
Recommendable. Very clean and taken care of with the necessary accessories.Manuel2019-07-31T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Excellent address that I recommend to travelers on Angers. Friendly and courteous host, very good accommodation.Jean-Marc2020-02-11T00:00:00Z
A warm welcome and a very pleasant setting!Cécile2020-02-09T00:00:00Z
Thank you for this excellent welcome. I highly recommend this quiet place in Angers!Aurelie2020-01-21T00:00:00Z
Nicolas is hospitable and friendlyYann2020-01-10T00:00:00Z
The place is convenient to reach from the station. There's a bus line 1 50 meters away from the house. Warm and clean room. With thoughtful breakfast. Very quite family!Grace2019-12-23T00:00:00Z
A very good welcome. Bed and breakfast to recommend!Martine2019-12-12T00:00:00Z
Close to the city center, a stone's throw from the St Nicolas pond, Nicolas' spacious and comfortable house won us over. We had a great stay with our 2 year old daughter in peace.Elise2019-12-11T00:00:00Z
Very nice house and perfectly clean. Very welcome Nicolas, I recommend.Lucie2019-10-16T00:00:00Z
Private room
Angers, Pays de la Loire, France
Just perfect. Magali is really very kind and attentive. The house is great, the room is large and comfortable. I had a great evening at Magali's!Chloé2020-03-07T00:00:00Z
Arranging and friendly. Magalie offered me a moment beyond my expectations! I highly recommendThomas2020-03-03T00:00:00Z
Magali was very accommodating and reactive. I recommendsimon2020-02-23T00:00:00Z
Very kindFlavien2020-01-10T00:00:00Z
Charming apartment Magali is friendly I recommendSamuel Branda2019-12-21T00:00:00Z
A very good address, well located, clean and above all with a warm welcome. A quality that we sometimes forget but which is most important !!!Daniel2019-12-15T00:00:00Z
Very pleasant and spontaneous welcome. I felt comfortable from the start. Well located.Philippe2019-11-28T00:00:00Z
Super hote!!Natacha2019-11-07T00:00:00Z
Private room
Angers, Pays de la Loire, France
Housing according to the description. Good report / quality. Very clean accommodation. Syriane is very responsive and gives very good advice.Valérie2019-11-24T00:00:00Z
Perfect for my two night stay for a business trip. The apartment is in a large building, with parking on site, easily accessible. Syriane was very friendly, and the apartment has everything you need!Marcela2019-11-22T00:00:00Z
The apartment is in line with the announcement and Syriane is very responsive I recommendMalik2019-10-13T00:00:00Z
Superb studio well done cleanliness irreproachable and very calm thank you very much for your welcomeJerome2019-10-06T00:00:00Z
Thank you for your welcome Syriane! The apartment is great, very nice and cozy, located close to everything I needed during this short stay. I recommend without worries!Chloé2019-09-20T00:00:00Z
Apartment in quiet area. Very nice and functional. Friendly welcome by Syriane.Melane2019-09-15T00:00:00Z
As always, the accommodation is always clean, functional with all the necessary equipment. We feel very good. I recommendIsabelle2019-09-06T00:00:00Z
Studio according to its description. Impeccable cleanliness. Friendly hostess. Good value for moneyMartine2019-08-13T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
A very warm welcome, a clean and well decorated room! Access to downtown Angers in 1 minute! I highly recommend.Andréi2020-03-08T00:00:00Z
Thank you for the night spent at the feet of the church. Being awakened by the sun and the gargoyles is great, I will come back with great pleasure and I highly recommend. In addition the hosts are very kind, attentive, accommodating and attentive. Confidence reigns and it's very pleasant.Pauline2020-03-03T00:00:00Z
Apparently atypical charming, perfect for a short stay. Ideally located for exploring Angers on foot (free parking 10 minutes on foot). Hugo and Nathan were reactive and allowed an easy organization. Thank you!Amandine2020-02-26T00:00:00Z
Excellent pied à terre for a weekend in Angers: ideally located, impeccable cleanliness, very accommodating hosts. Only one downside: the bed is really very soft, to be avoided for fragile backs!Marie2020-02-24T00:00:00Z
Very well placed. Complies with the desc. Have a good stayBastien2020-02-16T00:00:00Z
Great apartment right in the centre of AngersJo2020-02-01T00:00:00Z
Apartment not too noisy and meets expectations! Good stay spent in this superb accommodation.Jeanne2020-01-18T00:00:00Z
My daughter and I had a lovely stay! We appreciated the stylish details, check in was easy, and Angers is absolutely beautiful. We had no complaints and recommend this place.Xana2020-01-12T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Clean, pleasant accommodation, very consistent with the description, great stay and Brandon and available when needed Accommodation I strongly recommendClément2020-03-07T00:00:00Z
Super apartment. Ideally located in Angers. Contemporary decoration. I recommendPaul2020-02-15T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Warm welcome and on time. Apartment well placed (near the tram), clean, well equipped and well decorated. I will definitely go back. To recommend.Mehdi2020-03-14T00:00:00Z
Very quiet and clean apartment. In the top !Gerald2020-03-12T00:00:00Z
Clean apartment, well organized and close to transport to access the city center. Charline and her companion are pleasant hosts that I recommend.Justine2020-02-21T00:00:00Z
New apartment, with all amenities. Located on the street, but without overnight passage. Just the continuous breath of the VMC to review as much as possible.Nicolas2020-02-04T00:00:00Z
Great stay, super clean apartment well equipped! I advise !Éva2019-12-10T00:00:00Z
Have a nice stay at Charline. Very responsive to my messages, I recommend.David2019-12-01T00:00:00Z
Super housing of reactive hosts. Nothing to say except that I recommend.Natacha2019-11-29T00:00:00Z
A very charming apartment and well placed, I had a great stay and I recommend it and next time without a doubtLouise2019-11-24T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Jonathan is a high quality host, his apartment with exceptional facilities is tastefully decorated. We appreciated the little touches like coffee and candles.Delphine2020-03-16T00:00:00Z
Amazing stay in this unique place! Everything is wonderful, Jonathan is attentive and accessible.Elsa2020-03-13T00:00:00Z
Ideal for a romantic evening!Arnaud Macé2020-03-01T00:00:00Z
Thank you for this great weekend, we had a great time!Fanny2020-02-29T00:00:00Z
The apartment is very chic, perfectly equipped and very well located. We had a great time.Valerie2020-02-28T00:00:00Z
The apartment is even bigger in reality, very clean, no complaints we had a very nice stay I obviously recommendEloïse2020-02-26T00:00:00Z
Amazing place with a great location! All the amenities were amazing. We would definitely stay again.Charles2020-02-21T00:00:00Z
We had a wonderful stay of 3 nights and 4 days. The equipment on offer was better than the photo. A very spacious and functional jacuzzi. The apartment with a separate entrance allowed us to have our privacy. The little touches (fruit, drinks and coffee capsules) made our stay even more memorable. A dream apartment with quality service. To do again!Arshath2020-02-19T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Great stay in Angers! Your "cozy" is well laid out, a real cocoon where everything is thought out to make you feel goodTifany2020-01-26T00:00:00Z
Apartment very well placed and really consistent with the description. In addition Elise was able to show flexibility following an unforeseen on my part. Thank you so much! We had a pleasant stay in this apartment with girlfriends! Go for it!Magalie2020-01-12T00:00:00Z
The studio is very functional, super well placed. Good communication. I recommend!Laure2020-01-10T00:00:00Z
Comfortable accommodation, super well placed, clean and great welcome!Veronique2020-01-05T00:00:00Z
Apartment very well placed in the city center. Calm because it overlooks the courtyard. Élise is reactive and friendly!Celine2019-12-25T00:00:00Z
Thank you for your welcome and your availability. Apparently very well placed and tastefully optimized. See you soonHelene2019-11-16T00:00:00Z
Nothing to say in terms of location, decoration, equipment and communication with guests! Very nice stay at Elise and RomainSimon2019-11-08T00:00:00Z
Excellent stay in Angers in this beautiful apartment Everything went very well ! I recommend this addressRoxane2019-11-01T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Clean, practical accommodation and a really good jacuzzi for relaxing. A beautiful apartment very well locatedCeline2020-03-02T00:00:00Z
Alexandre's apartment is a real love nest in the heart of Angers. Close to all amenities, the hyper center and all the tourist places that you can do on foot. The decor and the jacuzzi were top notch. I recommend to all lovers who want to spend a romantic Angevin weekend!Lola2020-02-09T00:00:00Z
Exceptional !! Perfect accommodation, very clean, neat decor and very well equipped. Perfect location, close to everything. We were delighted with this stay.Julien2020-02-02T00:00:00Z
Perfect accommodation for a romantic getaway. Ideally located in Angers, tastefully decorated and very decent jacuzzi to relax ... Nothing to say, I keep this superb address for next time! Thank youGwladys2019-12-27T00:00:00Z
Very nice surprise when arriving at the accommodation. Ideal for a romantic eveningCyril2019-12-20T00:00:00Z
Fabulous apartment! The jacuzzi and the walk-in shower are a real bonus. Arrival in places very simple. Great location. Well soundproofed. Perfect value for money. I recommend with great pleasure :-)Quentin2019-12-18T00:00:00Z
Nice apartment and ideally located. I recommend.Jerome2019-12-15T00:00:00Z
Apartment at the height of the photos, a real cozy little nest. The decoration is really cozy, the most of this apartment the jacuzzi a treat for maximum relaxation. The apartment is also very well located. We did not see our host but more than clear and perfect instruction. thanks againSandra2019-12-11T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
Small, brand new studio apartment, superbly furnished and well placed for visiting the city.Thomas2020-01-12T00:00:00Z
Used to booking Mathieu's apartment each time I come to Angers, this time I was pleased to discover his new accommodation. Everything is spotless as usual! as in the previous apartment! Very nice and comfortable. I hope to be able to come back soon!Charline2020-01-11T00:00:00Z
Housing refurbished and very well equipped. Well located in a quiet street near the train station. The city center is very accessible on foot.Jean-Marc2019-12-30T00:00:00Z
I highly recommend this studio. Very clean, quiet, bright, well-kept, Angers center on foot. Matthieu is very responsive!Isabelle2019-12-24T00:00:00Z
Very clean apartment, very quiet, very well placed, perfect welcomeElisa2019-12-23T00:00:00Z
Nice accommodation in a good location. Nothing to say, the accommodation conforms to the photos.Sarafina2019-12-20T00:00:00Z
Perfect! Good communication. The location next to the station is top notch. Team. Quiet accommodation.Benoist2019-12-16T00:00:00Z
Mathieu is a very attentive and very friendly host. We had a great stay in his apartment which happens to be both practical and very cozy. I highly recommend it!Mickaël2019-12-15T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
This is a lovely little place to stay. Centrally located , really easy city to get to and to navigate your way round. solene is so easy to deal with too. Really recommend this to any couple, or single wanting to visit this beautiful and interesting cityLawrence2020-02-29T00:00:00Z
Very well located in Angers, clean accommodation, and good welcome from SolèneBrice2020-02-23T00:00:00Z
Very clean apartment. Warm and friendly welcome. I appreciated the little touches made available for breakfast.Karine2020-02-12T00:00:00Z
Super practical and comfortable, well placed and quiet, a house that can only please!Pierre2020-01-25T00:00:00Z
Maisonette perfectly conforms to the description, with all the necessary equipment, and very clean. Warm and friendly welcome from Solène. I highly recommend !Laurine2020-01-18T00:00:00Z
impeccable cleanliness, quiet apartment and at the same time not far from the center. I highly recommend it.Benoît2020-01-17T00:00:00Z
Solène's accommodation is pleasant and very well located. The welcome is warm and when we open the cupboards, we have the pleasure of discovering some cool products (coffee, tea, cracottes, jams ...) Very good local addresses (bars, restaurants). I recommend !Audrey2019-12-31T00:00:00Z
Solène offers you a house that will not leave you indifferent! Modern, functional and ideally placed, this home can only suit you!Arnold2019-12-15T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France
The welcome and the accommodation are just perfect.Vincent2020-02-07T00:00:00Z
Super welcome. Helpful and attentive people who put everything at your disposal. We don't find much service like thisAbdellah2020-01-29T00:00:00Z
Private room
Angers, Pays de la Loire, France
Very welcoming and very available hostBéatrice 2020-02-23T00:00:00Z
I advise this studio very wellJohan2020-01-05T00:00:00Z
Very welcome, Jérome strived to make our stay as perfect as possible. I recommend !Oceane2020-01-01T00:00:00Z
Great stay at Jérome's accommodation was clean and pleasantJuliette2019-12-20T00:00:00Z
Appart that keeps its promises. Clean, nicely decorated, quiet and close to the city center and shopsJean Noel2019-12-15T00:00:00Z
Everything is in accordance with the announcement, do not hesitate to book! thank you Jérôme for this very pleasant stay!Joan2019-08-28T00:00:00Z
Very good stay in one of the three studios of Jérôme. The room is comfortable with small, practical and new kitchen, and the shower is large and modern. Jerome is very responsive and available. The city center is 5 minutes walk. We recommend !Sarah2019-08-25T00:00:00Z
Very good stay, impeccable cleanliness and close to the centerLorraine2019-08-19T00:00:00Z
Entire home/apt
Angers, Pays de la Loire, France


text:
Getting in on a web meme a few billion years late, I just visited NameVoyager and started poking around to see trends in names. My name? Not even in the top 1000 baby names (for boys or girls) in 2003. Not with any spelling. It peaked in the 70's, being 174th most popular name for girls, 204th for boys. I'm shocked by that. I know very few people named Jody (as opposed to people using Jody as a nickname.)
My sister's name was the *7th* most popular girls name of the 50s! It had dropped all the way to 23rd by the 70s, and now her spelling, like mine, isn't even a blip on the radar. So I don't need any smart comments from her.
Just for fun, "Junior" ranks higher than "Jody." Not as high as his real name, but I'm fascinated that there are people out there actually giving a wee bitty baby the legal first name "Junior." It boggles.