Bladder cancer (BC) management demands the introduction of novel molecular targets for precision medicine. Cell surface glycoprotein CD44 has been widely studied as a potential biomarker of BC ...aggressiveness and cancer stem cells. However, significant alternative splicing and multiple glycosylation generate a myriad of glycoproteoforms with potentially distinct functional roles. The lack of tools for precise molecular characterization has led to conflicting results, delaying clinical applications. Addressing these limitations, we have interrogated the transcriptome and glycoproteome of a large BC patient cohort for splicing signatures.
gene and its splicing variants were assessed by Real Time-Polymerase Chain Reaction (RT-PCR) and RNAseq in tumor tissues. The co-localization of CD44 and short
-glycans was evaluated by proximity ligation assay (PLA), immunohistochemistry and double-immunofluorescence. An innovative glycoproteogenomics approach, integrating transcriptomics-customized datasets and glycomics for protein annotation from nanoLC-ESI-MS/MS experiments, was developed and implemented to identify CD44 variants and associated glycosignatures. The impact of CD44 silencing on proliferation and invasion of BC cell lines and glycoengineered cells was determined by BrdU ELISA and Matrigel invasion assays, respectively. Antibody phosphoarrays were used to investigate the role of CD44 and its glycoforms in the activation of relevant oncogenic signaling pathways.
Transcriptomics analysis revealed remarkable CD44 isoforms heterogeneity in bladder cancer tissues, as well as associations between short CD44 standard splicing isoform (CD44s), invasion and poor prognosis. We further demonstrated that targeting short
-glycoforms such as the Tn and sialyl-Tn antigens was key to overcome the lack of cancer specificity presented by CD44. Glycoproteogenomics allowed, for the first time, the comprehensive characterization of CD44 splicing code at the protein level. The concept was applied to invasive human BC cell lines, glycoengineered cells, and tumor tissues, enabling unequivocal CD44s identification as well as associated glycoforms. Finally, we confirmed the link between CD44 and invasion in CD44s-enriched cells
by small interfering RNA (siRNA) knockdown, supporting findings from BC tissues. The key role played by short-chain
-glycans in CD44-mediated invasion was also demonstrated through glycoengineered cell models.
Overall, CD44s emerged as biomarker of poor prognosis and CD44-Tn/ Sialyl-Tn (STn) as promising molecular signatures for targeted interventions. This study materializes the concept of glycoproteogenomics and provides a key vision to address the cancer splicing code at the protein level, which may now be expanded to better understand CD44 functional role in health and disease.
Primeiro registro de Lutzomyia araracuarensis (Morales & Minter) (Diptera: Psychodidae) no Brasil Silva, Dílvia F da(Univ Federal Rural de Pernambuco Lab de Biologia dos Insetos); Freitas, Rui A de(Instituto Nacional de Pesquisas da Amazônia Lab de Leishmaniose e Doença de Chagas); Franco, Antonia M R(Instituto Nacional de Pesquisas da Amazônia Lab de Leishmaniose e Doença de Chagas)
Neotropical entomology,
10/2010, Letnik:
39, Številka:
5
Journal Article
Recenzirano
Odprti dostop
A male of Lutzomyia araracuarensis (Morales & Minter) and possibly six females of this same species were found in the northeastern area of Manacapuru county, Amazonas State. Samples were collected ...using light traps CDC, from April 2003 to June 2004.
Purpose
The most frequent histology of bladder tumors is urothelial carcinoma. Most are pure urothelial carcinomas (PUC) but up to one-third of the cases present variant histological (VH) features. ...The aim of this study was to evaluate the role of variant histology in neoadjuvant chemotherapy (NAC) response in patients with urothelial muscle-invasive bladder cancer.
Methods
We retrospectively analyzed data from 77 patients with bladder cancer who performed neoadjuvant chemotherapy at two institutions.
Results
Complete pathological response (ypT0) was higher in patients with PUC (38.5%), comparing with VH (12%). Logistic regression analysis demonstrated that variant histology is associated with an 89% lesser likelihood of tumor downstaging, with advanced clinical T stages and positive smoking history as independent predictors. The estimated mean cancer-specific survival was 68.91 months for PUC patients and 50.23 months for VH patients (log rank test,
P
= 0.024). Multivariate Cox regression analysis demonstrated that VH and clinical T stage were independent predictors of cancer-specific survival, indicating a worse outcome for patients with VH and advanced clinical T stages.
Conclusions
There are only a few retrospective studies evaluating the clinical impact of variant histology tumors, which are mainly managed as PUC. Our results demonstrate that VH is associated with a worse likelihood of tumor downstaging after NAC and a worse cancer-specific survival in bladder cancer patients. There is a need for further studies and genetic analysis to identify the patients most likely to achieve ypT0 status and downstaging after NAC.
Abstract
The common dolphinfish (Coryphaena hippurus) is an epipelagic, mid-trophic level, highly migratory species distributed throughout the world’s tropical and subtropical oceans in waters ...greater than 20°C. Life-history variables, migratory behaviour, and genetic markers have been used to define major stocks in the central Atlantic Ocean and Mediterranean Sea. Here, we used the mitochondrial DNA gene NADH subunit 1 (688 bp) to test for differences between population groups. A total of 103 haplotypes were detected among 203 fish. Gene diversities in samples were large and similar among populations (mean h = 0.932; range 0.894–0.987), but nucleotide diversities varied widely among samples (range π = 0.004–0.034) and appear to reflect population histories. Principal component analysis revealed two large populations groups, and the analysis of molecular variation and pairwise values of ΦST resolved population structure within these groups. Populations in the eastern Atlantic and Mediterranean showed the largest amounts of divergence from one another (ΦCT = 0.331). Adult movement and biophysical barriers to larval dispersal may explain contemporary differences between stocks, but the divergent populations in the Mediterranean Sea are likely due to isolations by cold temperature barriers during Pleistocene glaciations. The geographically large stock groupings require international cooperation in the harvest management and conservation of local dolphinfish populations.
To evaluate the potential of sialyl-Tn (STn), a cancer-associated glycan antigen present in membrane glycoproteins, to improve a recent molecular model for stratification and prognostication of ...advanced stage bladder tumors based on keratins (KRT14, 5, and 20) expression. In addition, determine the association between STn and disease dissemination based on the evaluation of circulating tumor cells (CTCs) and the metastasis, which is a critical matter to improve patient management.
A retrospective series of 80 muscle-invasive primary bladder tumors and associated metastasis were screened for KRT14, 5, and 20 and STn by real-time polymerase chain reaction and immunohistochemistry. Peripheral blood was collected in a patients’ subset, CTCs were isolated through a size-based microfluidic chip and screened for KRTs and STn.
Basal-like lesions presented worse cancer-specific and disease-free survival compared to luminal tumors. STn antigen inclusion discriminated patients with worst survival in each subgroup (P = 0.047 for luminal; P = 0.027 for basal-like tumors). STn expression in CTCs and distant metastasis was also demonstrated.
This work reinforces the potential of the KRT-based model for bladder cancer management and the association of STn with aggressiveness, supporting its inclusion in predictive molecular models toward patient-tailored precision medicine. Moreover, we describe for the first time that CTCs and the metastasis present a basal phenotype and express the STn antigen, highlighting its link with disease dissemination. Future studies should focus on determining the biological and clinical significance of these observations in the context of liquid biopsies. Given the membrane nature of STn, highly specific targeted therapeutics may also be envisaged.
•Basal-like lesions presented worse survival compared to luminal tumours.•Sialyl-Tn inclusion in KRT-based model discriminated patients with worst survival.•Sialyl-Tn is expressed in circulating tumour cells and distant metastasis.•Sialyl-Tn has potential to be a novel biomarker for targeted therapeutics.
Declines in Panulirus echinatus Smith, 1869 populations along their wide distribution in the Atlantic Ocean have spurred efforts to improve their fisheries management and conservation. In this study, ...the genetic structure of these populations is reported for the first time.
In a survey of 18 species‐specific polymorphic microsatellite markers, 152 individuals were genotyped from five Atlantic oceanic islands, covering most of the insular distribution range of the species. The analyses revealed that P. echinatus is genetically partitioned into two stocks in the Atlantic Ocean. A highly significant genetic structure was observed between north‐east and south‐west Atlantic populations based on fixation index, discriminant analysis of principal components, and structure and barrier analysis.
We suggest that the Equatorial Circulation System represents a biophysical barrier that effectively limits migration among Atlantic subtropical gyre systems, as has been described for other species. Other physical and ecological barriers, such as the Mid‐Atlantic Ridge itself, the distance between the eastern and western sides of the Atlantic (Mid‐Atlantic Barrier) and water mass differences, as well as other biological aspects, may also influence larval dispersal and modulate the insular distribution of this species.
The results show the existence of two distinct genetic stocks of P. echinatus and have implications for fisheries management in the Atlantic Ocean, including their independent management according to their individual status. The Cabo Verde and the Canarian populations (north‐east Atlantic) showed the lowest level of genetic variability in comparison with the south‐western populations. A combination of factors that have occurred or are occurring in the Canary Islands, such as overfishing and volcanic eruptions, is likely to explain the reduced abundance of this lobster species in the area.
Summary Objectives/Hypothesis The most recent and significant magnetic resonance imaging (MRI) improvements allow for the visualization of the vocal tract during speech production, which has been ...revealed to be a powerful tool in dynamic speech research. However, a synchronization technique with enhanced temporal resolution is still required. Study Design and Methods The study design was transversal in nature. Throughout this work, a technique for the dynamic study of the vocal tract with MRI by using the heart's signal to synchronize and trigger the imaging-acquisition process is presented and described. The technique in question is then used in the measurement of four speech articulatory parameters to assess three different syllables (articulatory gestures) of European Portuguese Language. Results The acquired MR images are automatically reconstructed so as to result in a variable sequence of images (slices) of different vocal tract shapes in articulatory positions associated with Portuguese speech sounds. Conclusions The knowledge obtained as a result of the proposed technique represents a direct contribution to the improvement of speech synthesis algorithms, thereby allowing for novel perceptions in coarticulation studies, in addition to providing further efficient clinical guidelines in the pursuit of more proficient speech rehabilitation processes.
Lutzomyia georgii n. sp. and the female of L. tarapacaensis in the
Series infraspinosa of the subgenus Evandromyia are described, from
specimens collected in rainforest in the north of the State of
...Pará, Brazil. The new species was taken together with five other
Evandromyia species including L. infraspinosa (sensu strictu) in the
same locality. L. georgii has previously been confused with both L.
begonae and L. infraspinosa, whereas L. tarapacaensis would run to L.
infraspinosa in recent taxonomic keys. The fact that both L. georgii
and L. tarapacaensis are locally sympatric with L. infraspinosa helps
to clarify the taxonomic limits of the latter species. New keys to the
subgenus Evandromyia are provided.
Prevalence of Visual Impairment in Portugal Carneiro de Freitas, Rui; Freitas, Cristina; Leite, Ricardo ...
Investigative ophthalmology & visual science,
2015, Letnik:
56, Številka:
7
Journal Article
Recenzirano
Purpose
Information about the prevalence of visual impairment is fundamental to define policies that deal with vision loss. The aim of this study is to determine the prevalence of visual impairment ...(VI) in the population looking for eye care in public hospitals in Portugal.
Methods
We designed an observation, cross-sectional prospective study (Prevalence and Costs of Visual Impairment in Portugal: PC-VIP study) to investigate the prevalence of VI in patients attending outpatient appointments in four public hospitals in Portugal. Hospital selected provide from general eye care (3-6 ophthalmologists) to high-specialized eye care (40+ ophthalmologists) that in total have between 120-140K hospital appointments per year. Files of patients are analysed weekly to detect patients with VI. Inclusion criteria were: visual acuity equal or worse than 0.5 (USA definition 20/40) in the better eye and/or a visual field of less than 20deg. Cases are selected by trained hospital staff and inserted in a database. Data collected included demographic information, acuity from both eyes, qualitative information about visual field (good, reduced, requires investigation), main diagnosis, secondary diagnosis and comorbidities. Diagnoses were classified according with ICD9.
Results
We have now detected 2462 cases of VI that correspond to 4 to 33 weeks of data collection. The number of weeks is variable because collection did not start simultaneously in all sites. From the number of cases detected, 58% were female, 1.9% were under 20y, 8.2% were between 20y and 50y and 89.9% were ≥50y. The mean prevalence of visual impairment was 13.6% (SD=5.6) using the USA definition and it was 7.0%(SD=4.1) using the WHO definition (acuity equal or worse than 0.3 or ~20/63). With a methodology that controls for demographics the lowest and highest estimates were calculated. Considering the USA definition, the prevalence in the general population would be in the range 0.4 -0.4% (age<40y) and 0.8-2.4% (age>=40y). Considering WHO definition, it would be 0.2-0.5% (age<40y) and 0.4-1.0% (age>=40y).
Conclusions
A hospital-based study can provide effective estimates of the prevalence of visual impairment in a population. Estimates for the country are in agreement with the expected results that can be deducted from neighbour countries and self-reported visual impairment in census 2001.
PDF
