An inducible CiliaGFP mouse model for in vivo visualization and analysis of cilia in live tissue

BACKGROUND: Cilia are found on nearly every cell type in the mammalian body, and have been historically classified as either motile or immotile. Motile cilia are important for fluid and cellular movement; however, the roles of non-motile or primary cilia in most tissues remain unknown. Several genetic syndromes, called the ciliopathies, are associated with defects in cilia structure or function and have a wide range of clinical presentations. Much of what we know about the formation and maintenance of cilia comes from model systems like C. elegans and Chalmydomonas. Studies of mammalian cilia in live tissues have been hampered by difficulty visualizing them. RESULTS: To facilitate analyses of mammalian cilia function we generated an inducible Cilia(GFP) mouse by targeting mouse cDNA encoding a cilia-localized protein somatostatin receptor 3 fused to GFP (Sstr3::GFP) into the ROSA26 locus. In this system, Sstr3::GFP is expressed from the ubiquitous ROSA26 promoter after Cre mediated deletion of an upstream Neo cassette flanked by lox P sites. Fluorescent cilia labeling was observed in a variety of live tissues and after fixation. Both cell-type specific and temporally regulated cilia labeling were obtained using multiple Cre lines. The analysis of renal cilia in anesthetized live mice demonstrates that cilia commonly lay nearly parallel to the apical surface of the tubule. In contrast, in more deeply anesthetized mice the cilia display a synchronized, repetitive oscillation that ceases upon death, suggesting a relationship to heart beat, blood pressure or glomerular filtration. CONCLUSIONS: The ability to visualize cilia in live samples within the Cilia(GFP) mouse will greatly aid studies of ciliary function. This mouse will be useful for in vivo genetic and pharmacological screens to assess pathways regulating cilia motility, signaling, assembly, trafficking, resorption and length control and to study cilia regulated physiology in relation to ciliopathy phenotypes

Host-guest and guest-guest interactions between xylene isomers confined in the MIL-47(V) pore system

The porous MIL-47 material shows a selective adsorption behavior for para-, ortho-, and meta-isomers of xylenes, making the material a serious candidate for separation applications. The origin of the selectivity lies in the differences in interactions (energetic) and confining (entropic). This paper investigates the xylene-framework interactions and the xylene-xylene interactions with quantum mechanical calculations, using a dispersion-corrected density functional and periodic boundary conditions to describe the crystal. First, the strength and geometrical characteristics of the optimal xylene-xylene interactions are quantified by studying the pure and mixed pairs in gas phase. An extended set of initial structures is created and optimized to sample as many relative orientations and distances as possible. Next, the pairs are brought in the pores of MIL-47. The interaction with the terephthalic linkers and other xylenes increases the stacking energy in gas phase (-31.7 kJ/mol per pair) by roughly a factor four in the fully loaded state (-58.3 kJ/mol per xylene). Our decomposition of the adsorption energy shows various trends in the contributing xylene-xylene interactions. The absence of a significant difference in energetics between the isomers indicates that entropic effects must be mainly responsible for the separation behavior

Tear fluid biomarkers in ocular and systemic disease: potential use for predictive, preventive and personalised medicine

In the field of predictive, preventive and personalised medicine, researchers are keen to identify novel and reliable ways to predict and diagnose disease, as well as to monitor patient response to therapeutic agents. In the last decade alone, the sensitivity of profiling technologies has undergone huge improvements in detection sensitivity, thus allowing quantification of minute samples, for example body fluids that were previously difficult to assay. As a consequence, there has been a huge increase in tear fluid investigation, predominantly in the field of ocular surface disease. As tears are a more accessible and less complex body fluid (than serum or plasma) and sampling is much less invasive, research is starting to focus on how disease processes affect the proteomic, lipidomic and metabolomic composition of the tear film. By determining compositional changes to tear profiles, crucial pathways in disease progression may be identified, allowing for more predictive and personalised therapy of the individual. This article will provide an overview of the various putative tear fluid biomarkers that have been identified to date, ranging from ocular surface disease and retinopathies to cancer and multiple sclerosis. Putative tear fluid biomarkers of ocular disorders, as well as the more recent field of systemic disease biomarkers, will be shown

pMel17 is recognised by monoclonal antibodies NKI-beteb, HMB-45 and HMB-50 and by anti-melanoma CTL.

Recently, we cloned the cDNA encoding the melanocyte lineage-specific antigen gp100 and demonstrated that gp100 is recognised by three different monoclonal antibodies (MAbs) used to diagnose malignant melanoma. In addition, we showed that tumour-infiltrating lymphocytes (TIL 1200) from a melanoma patient reacted specifically with cells transfected with the gp100 cDNA. Molecular characterisation of the gp100 cDNA revealed that the gp100 antigen is highly homologous, but not identical, to another melanocyte-specific protein, pMel17. Here, we report that cells transfected with pMel17 cDNA also react with all three MAbs used to diagnose malignant melanoma, NKI-beteb, HMB-45 and HMB-50. Moreover, pMel17 transfectants are specifically lysed by TIL1200. These data demonstrate that antigenic processing of both gp100 and pMel17 give rise to peptides seen by anti-melanoma cytotoxic T lymphocytes (CTL) and are therefore potential targets for immunotherapy of malignant melanoma

Parafoveal preview effects from word N+1 and word N+2 during reading: A critical review and Bayesian meta-analysis

The use of gaze-contingent display techniques to study reading has shown that readers attend not only to the currently fixated word, but also to the word to the right of the current fixation. However, a critical look at the literature shows that there are a number of questions that cannot be readily answered from the available literature reviews on the topic. First, there is no consensus on whether readers also attend to the second word to the right of fixation. Second, it is not clear whether parafoveal processing is more efficient in languages such as Chinese. Third, it is not well understood whether the measured effects are confounded by the properties of the parafoveal mask. The present study addressed these issues by performing a Bayesian meta-analysis of 93 experiments that used the boundary paradigm (Rayner, 1975). There were three main findings: 1) the advantage of previewing the second word to the right is modest in size and likely not centred on zero; 2) Chinese readers seem to make a more efficient use of parafoveal processing, but this is mostly evident in gaze duration; 3) there are interference effects associated with using different parafoveal masks that roughly increase when the mask is less word-like

Expanding the diversity of mycobacteriophages: Insights into genome architecture and evolution

Mycobacteriophages are viruses that infect mycobacterial hosts such as Mycobacterium smegmatis and Mycobacterium tuberculosis. All mycobacteriophages characterized to date are dsDNA tailed phages, and have either siphoviral or myoviral morphotypes. However, their genetic diversity is considerable, and although sixty-two genomes have been sequenced and comparatively analyzed, these likely represent only a small portion of the diversity of the mycobacteriophage population at large. Here we report the isolation, sequencing and comparative genomic analysis of 18 new mycobacteriophages isolated from geographically distinct locations within the United States. Although no clear correlation between location and genome type can be discerned, these genomes expand our knowledge of mycobacteriophage diversity and enhance our understanding of the roles of mobile elements in viral evolution. Expansion of the number of mycobacteriophages grouped within Cluster A provides insights into the basis of immune specificity in these temperate phages, and we also describe a novel example of apparent immunity theft. The isolation and genomic analysis of bacteriophages by freshman college students provides an example of an authentic research experience for novice scientists. © 2011 Hatfull et al

Conversational correlates of children's acquisition of mental verbs and a theory of mind

The purpose of this study was to conduct a detailed examination of the ways mothers use mental verbs in conversations with three- and four-year-old children, and to link these usages to the children's developing understanding of mental verbs and a theory of mind. Sixty three- and four-year-olds, either attending preschool (PS) or not (NPS) were given tasks assessing mental verb distinctions and false belief. Their mothers' mental verb use was coded for (a) frequency, (b) type of utterance, (c) type of subordinate clause, (d) the person of the subject of the verb, and (e) the certainty of think. Within the three-year-olds, the NPS children performed significantly better on the mental verb comprehension task; moreover, compared to the PS mothers, the NPS mothers were found to use: (1) less statements and more questions, (2) less first person utterances and more second person utterances, and (3) think in its `very certain' form less often. In regression analyses, children's mental verb and false belief performance were positively predicted by maternal mental verb 1) questions, and 2) single clause utterances; the children's performance was negatively predicted by statements. These findings indicate how maternal input has the potential to promote or hinder children's understanding of the mind.Yeshttps://us.sagepub.com/en-us/nam/manuscript-submission-guideline

Le ricerche preistoriche dell'Università di Ferrara

Il volume costituisce la sintesi scientifica delle ricerche preistoriche che l’Università degli Studi di Ferrara ha effettuato negli ultimi 50 anni, coprendo un arco temporale che comprende culturalmente il periodo che va dal Paleolitico inferiore al Mesolitico. Nel tentativo di fornire un quadro quanto più completo ed esaustivo delle ricerche la monografia è concepita come un catalogo di agile consultazione suddiviso in base ai siti archeologici scavati dai professori e ricercatori nel corso del tempo. Per ciascun sito si descrivono molteplici aspetti, dalle modalità di intervento allo studio di vari settori disciplinari (stratigrafia, geologia, palinologia, paleontologia, paletnologia, ecc..)

Correlation of enhanced thrombospondin-1 expression, TGF-β signalling and proteinuria in human type-2 diabetic nephropathy

Background. Activation of the thrombospondin-1 (TSP-1)-TGF-β pathway by glucose and the relevance of TSP-1-dependent activation of TGF-β for renal matrix expansion, renal fibrosis and sclerosis have previously been demonstrated by our group in in vivo and in vitro studies

SGLT2-Inhibition in Patients With Alport syndrome

\ua9 2024 International Society of NephrologyIntroduction: Large-scale trials showed positive outcomes of sodium-glucose cotransporter-2 inhibitors (SGLT2i) in adults with chronic kidney disease (CKD). Whether the use of SGLT2i is safe and effective in patients with the common hereditary CKD Alport syndrome (AS) has not yet been investigated specifically in larger cohorts. Methods: This observational, multicenter, international study (NCT02378805) assessed 112 patients with AS after start of SGLT2i. The study\u27s primary end point was change of albuminuria in albumin/g creatinine from the start of therapy. Results: Compared to randomized trials investigating the effect of SGLT2i in CKD, the adult patients in this study were younger (aged 38 \ub1 14 years) and had a better estimated glomerular filtration rate (eGFR, 63 \ub1 35 ml/min per 1.73 m2; n = 98). Maximum follow-up was 32 months. Compared to baseline, at the first 3 follow-up visits (months 1 to 3, 4 to 8, and 9 to 15) after initiation of SGLT2i therapy, a significant reduction of albuminuria in mg albumin/g creatinine (>30%) was observed. Mean loss of eGFR was 9 \ub1 12 ml/min per 1.73 m2 almost 1 year after initiation of SGLT2i therapy (n = 35). At a total of 71 patient-years at risk, 0.24 adverse events (AEs) per patient-year on SGLT2i were reported. Conclusion: This study indicates that, additive to renin-angiotensin system (RAS)-inhibition (RASi), SGLT2i have the potential to reduce the amount of albuminuria in patients with AS. Future studies are needed to investigate the long-term effects of SGLT2i on CKD progression in patients with AS to assess whether the observed reduction in albuminuria translates to a delay in kidney failure (KF)