The Caatinga biome is an important ecosystem in the semi-arid region of Brazil.It has significantly degraded due to human activities and is currently a region undergoing desertification.Thus,monitoring the variation i...The Caatinga biome is an important ecosystem in the semi-arid region of Brazil.It has significantly degraded due to human activities and is currently a region undergoing desertification.Thus,monitoring the variation in the Caatinga biome has become essential for its sustainable development.However,traditional methods for estimating aboveground biomass(AGB)are time-consuming and destructive.Remote sensing,such as optical and radar imaging,can estimate and correlate with vegetation.Nevertheless,radar imaging is still a novelty to be applied in estimating the AGB of this biome,which is an area with little research.Therefore,this study aimed to use Sentinel-1 images to estimate the AGB of the Caatinga biome in Sergipe State(northeastern Brazil)and to verify its influencing factors.Nineteen sample plots(30 m×30 m)were selected,and the stems of individuals with a circumference at breast height(1.3 m above the ground)equal to or greater than 6.0 cm were measured,and the AGB through an allometric equation was estimated.The Sentinel-1 images from 3 different periods(green,intermediate,and dry periods)were used to consider the phenological conditions of the Caatinga biome.All the pre-processing and extraction of attributes(co-polarized VV(vertical transmit and vertical receive),cross-polarized VH(vertical transmit and horizontal receive),and band ratio VH/VV backscatter,radar vegetation index,dual polarization synthetic aperture radar(SAR)vegetation index(DPSVI),entropy(H),and alpha angle(α))were performed with Sentinel’s Application Platform.These attributes were used to estimate the AGB through simple and multiple linear regressions and evaluated by the coefficients of determination(R2),correlation(r),and root mean squared error(RMSE).The results showed that the attributes individually had little ability to estimate the AGB of the Caatinga biome in the three periods.Combined with multiple regression,we found that the intermediate period presented the equation with the best results among the observed and estimated variables(R^(2)=0.73;r=0.85;RMSE=8.33 Mg/hm^(2)),followed by the greenness period(R2=0.72;r=0.85;RMSE=8.40 Mg/hm^(2)).The attributes contributing to these equations were VH/VV,DPSVI,H,α,and co-polarized VV for the green period and cross-polarized VH for the intermediate period.The study showed that the Sentinel-1 images could be used to estimate the AGB of the Caatinga biome in the green and intermediate phenological periods since the SAR attributes highly correlated with the estimated variable(i.e.,AGB)through multiple linear equations.展开更多
AIM: To compare gene expression profiles of pancreatic adenocarcinoma tissue specimens, human pancreatic and colon adenocarcinoma and leukemia cell lines and normal pancreas samples in order to distinguish differenti...AIM: To compare gene expression profiles of pancreatic adenocarcinoma tissue specimens, human pancreatic and colon adenocarcinoma and leukemia cell lines and normal pancreas samples in order to distinguish differentially expressed genes and to validate the differential expression of a subset of genes by quantitative real-time RT-PCR (RT-QPCR) in endoscopic ultrasound-guided fine needle aspiration (EUS-guided FNA) specimens.METHODS: Commercially dedicated cancer cDNA macroarrays (Atlas Human Cancer 1.2) containing 1176 genes were used. Different statistical approaches (hierarchical clustering, principal component analysis (PCA) and SAM) were used to analyze the expression data. RT-QPCR and immunohistochemical studies were used for validation of results.RESULTS: RT-QPCR validated the increased expression of LCN2 (lipocalin 2) and for the first time PLAT (tissue-type plasminogen activator or tPA) in malignant pancreas as compared with normal pancreas. Immunohistochemical analysis confirmed the increased expression of LCN2 protein localized in epithelial cells of ducts invaded by carcinoma. The analysis of PLAT and LCN2 transcripts in 12 samples obtained through EUS-guided FNA from patients with pancreatic adenocarcinoma showed significantly increased expression levels in comparison with those found in normal tissues, indicating that a sufficient amount of high quality RNA can be obtained with this technique.CONCLUSION: Expression profiling is a useful method to identify biomarkers and potential target genes. Molecular analysis of EUS-guided FNA samples in pancreatic cancer appears as a valuable strategy for the diagnosis of pancreatic adenocarcinomas.展开更多
AIM: TO study the implication of prokineticin 1 (PKI/EGVEGF) and prokineticin 2 (PK2/13v8) in hepatocellular carcinoma angiogenesis.METHODS: The gene induction of PK1/EG-VEGF and PK2/Bv8 was investigated in 10 n...AIM: TO study the implication of prokineticin 1 (PKI/EGVEGF) and prokineticin 2 (PK2/13v8) in hepatocellular carcinoma angiogenesis.METHODS: The gene induction of PK1/EG-VEGF and PK2/Bv8 was investigated in 10 normal, 28 fibrotic and 28 tumoral livers by using real time PCR. Their expression was compared to the expression of VEGF (an angiogenesis marker), vWF (an endothelial cell marker) and to CD68 (a monocyte/macrophage marker). Furthermore, the rnRNA levels of PK1/EG-VEGF, PK2/Bv8, prokineticin receptor 1 and 2 were evaluated by real time PCR in isolated liver cell populations. Finally, PK2/Bv8 protein was detected in normal liver paraffin sections and in isolated liver cells by immunohistochernistry and immunocytochemistry.RESULTS: PK2/Bv8 mRNA but not PK1/EG-VEGF was expressed in all types of normal liver samples examined. In the context of liver tumor development, we reported that PK2/13v8 correlates only with CD68 and showed a significant decrease in expression as the pathology evolves towards cancer. Whereas, VEGF and vWF mRNA were significantly upregulated in both fibrosis and HCC,as expected. In addition, out of all isolated liver cells examined, only Kupffer cells (liver resident macrophages) express significant levels of PK2/Bv8 and its receptors, prokineticin receptor 1 and 2.CONCLUSION: In normal liver PK2/Bv8 and its receptors were specifically expressed by Kupffer cells. PK2/Bv8 expression decreased as the liver evolves towards cancer and did not correlate with HCC angiogenesis.展开更多
AIM: TO investigate in vivo, whether CCK2 receptors (CCK2R) regulate proteins known to play a crucial role in cell proliferation and cancer development and analyse in vitro the molecular mechanisms that lead to Src...AIM: TO investigate in vivo, whether CCK2 receptors (CCK2R) regulate proteins known to play a crucial role in cell proliferation and cancer development and analyse in vitro the molecular mechanisms that lead to Src activation; in particular, to identify the domains within the CCK2R sequence that are implicated in this activation. METHODS: The expression and activation of Src and ERK were studied in vivo using immunofluorescence and western-blot techniques. We used pancreatic tissues derived from wild type or Elas-CCK2 mice that expressed the CCK2R in pancreatic acini, displayed an increased pancreatic growth and developed preneoplastic lesions. The pancreatic tumor cell line AR4-2J expressing the endogenous CCK2R or COS-7 cells transiently transfected with wild type or mutant CCK2R were used as in vitro models to study the mechanism of Src activation. Src activation was measured by in vitro kinase assays, ERK activation by western blot using antiphospho-ERK antibodies and the involvement of Src in gastrin-induced cell proliferation by MTT test. RESULTS: We showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, led to the activation of Src and the ERK pathway. Src was activated upstream of the ERK pathway by the CCK2R in pancreatic tumoral cells and contributed to the proliferative effects mediated by this receptor. In vitro results demonstrated that activation of the Src/ERK pathway by the CCK2R required the NPXXY motif, located within the CCK2R sequence at the end of the 7^th transmembrane domain, and suggested the putative role of Gq in this mechanism. CONCLUSION: Deregulation of the Src/ERK pathway by the CCK2R might represent an early step that contributes to cell proliferation, formation of preneoplastic lesions and pancreatic tumor development.展开更多
Acute humoral rejection (AHR) is uncommon after ABO- compatible liver transplantation. Herein, we report two cases of AHR treated with plasmapheresis and rituximab in two ABO-compatible liver-transplant patients wit...Acute humoral rejection (AHR) is uncommon after ABO- compatible liver transplantation. Herein, we report two cases of AHR treated with plasmapheresis and rituximab in two ABO-compatible liver-transplant patients with preformed anti-human leukocyte antigen donor-specific antibodies. Patient 1 experienced a biopsy-proven AHR at day 10 post-transplant. She was treated by steroid pulses, and OKT3. Because of persisting signs of biopsy-proven AHR at day 26, she was treated by plasmapheresis and rituximab. Uver enzyme levels did not improve, and she died on day 41. Patient 2 experienced a biopsy-proven AHR on day 10 post-transplant. She was treated by steroid pulses, plasmapheresis, and rituximab. Liver enzymes returned to within normal range 18 d after diagnosis. Uver biopsies, at 3 and 9 mo post-transplant, showed complete resolution of AHR. We conclude that plasmapheresis should be started as soon as AHR is diagnosed, and be associated with a B-cell depleting agent. Rituximab may be considered as a first-line therapy.展开更多
AIM: To evaluate the implication of substitutions in the hepatitis C virus (HCV) non-structural 5A (NS5A) protein in the resistance of HCV during mono-interferon (IFN) or combined IFN-ribavirin (IFN-R) therapy. Althou...AIM: To evaluate the implication of substitutions in the hepatitis C virus (HCV) non-structural 5A (NS5A) protein in the resistance of HCV during mono-interferon (IFN) or combined IFN-ribavirin (IFN-R) therapy. Although NS5A has been reported to interact with the HCV RNA- dependent RNA polymerase, NS5B, as well as with many cellular proteins, the function of NS5A in the life cycle of HCV remains unclear. METHODS: HCV quasispecies were studied by clon- ing and sequencing of sequential isolates from patients infected by HCV genotype 1b. Patients were treated by IFN-α2b for 3 mo followed by IFN-α2b alone or com- bined IFN-R therapy for 9 additional months. Patients were categorized intro two groups based on their re- sponse to the treatments: 7 with sustained virological re- sponse (SVR) (quasispecies = 150) and 3 non-respond- ers (NR) to IFN-R (quasispecies = 106). RESULTS: Prior to treatment, SVR patients displayed a lower complexity of quasispecies than NR patients. Most patients had a decrease in the complexity of quasispe- cies during therapy. Analysis of amino acids substitu- tions showed that the degree of the complexity of the interferon sensitivity-determining region (ISDR) and the V3 domain of NS5A protein was able to discriminate thetwo groups of patients. Moreover, SVR patients displayed more variability in the NS5A region than NR patients. CONCLUSION: These results suggest that detailed mo- lecular analysis of the NS5A region may be important for understanding its function in IFN response during HCV 1b infection.展开更多
AIM: To investigate whether Src, JAK2 and phosphatidylinositol 3-kinase (PI3K) pathways are involved in the proliferation of human colonic tumour cells induced by glycine-extended gastrin (G-gly), the precursor o...AIM: To investigate whether Src, JAK2 and phosphatidylinositol 3-kinase (PI3K) pathways are involved in the proliferation of human colonic tumour cells induced by glycine-extended gastrin (G-gly), the precursor of the mature amidated gastrin and to elucidate the molecular interaction between these three kinases in response to this peptide. METHODS: Using the human colonic tumour cell line HCT116 as a model, we first measured the activation of PI3K, p60-Src and JAK2 in response to G-gly by in vitro kinase assays. Then we investigated the involvement of these kinases in G-gly-induced cell proliferation by MTT test. RESULTS: G-gly stimulation induced p60-Src, JAK2 and PI3K activation in HCT116. The different pathways were involved in proliferation of human colon cancer cells induced by G-gly. Furthermore, we found that both Src and JAK2 were necessary to PI3K regulation by this peptide. However, we did not find any cross-talk between the two tyrosine kinases. CONCLUSION: Our results suggest that the p60-Src/ PI3K and JAK2/PI3K pathways act independently to mediate G-gly proliferative effect on human colonic tumour cells.展开更多
Salmonella can invade non-phagocytic cells through its type Ⅲ secretion system (T3SS-1), which induces a Trigger entry process. This study showed that Salmonella enterica, subspecies enterica serovar Enteritidis ca...Salmonella can invade non-phagocytic cells through its type Ⅲ secretion system (T3SS-1), which induces a Trigger entry process. This study showed that Salmonella enterica, subspecies enterica serovar Enteritidis can also invade cells via the Rck outer membrane protein. Rck was necessary and sufficient to enable non-invasive E. coli and Rckcoated beads to adhere to and invade different cells. Internalization analysis of latex beads coated with different Rck peptides showed that the peptide containing amino acids 140-150 promoted adhesion, whereas amino acids between 150 and 159 modulated invasion. Expression of dominant-negative derivatives and use of specific inhibitors demonstrated the crucial role of small GTPases Racl and Cdc42 in activating the Arp2/3 complex to trigger formation of actin-rich accumulation, leading to Rck-dependent internalization. Finally, scanning and transmission electron microscopy with Rck-coated beads and E. coli expressing Rck revealed microvillus-like extensions that formed a Zipper-like structure, engulfing the adherent beads and bacteria. Overall, our results provide new insights into the Salmonella T3SS-independent invasion mechanisms and strongly suggest that Rck induces a Zipper-like entry mechanism. Consequently, Salmonella seems to be the first bacterium found to be able to induce both Zipper and Trigger mechanisms to invade host cells.展开更多
The objective of this research is to present a systematic analysis, in which we seek to identify the concepts of intelligent city, with the intention of recognizing the nature of this intelligence before the participa...The objective of this research is to present a systematic analysis, in which we seek to identify the concepts of intelligent city, with the intention of recognizing the nature of this intelligence before the participation of the population. Therefore, the research associates concepts as smart cities, citizen participation and social web, identifying publications between 2012 and the first quarter of 2018 and questioning what the intelligent city is and how this concept is able to (re)organize the learning processes of the territory from the informal dynamics of the contemporary city. The qualitative analysis of the documents revealed an innumerable of definitions and related terms: smart, intelligent, ubiquitous, digital, knowledge, sustainable, crowd sourcing, innovative;structured in three types of approaches: technological focus, focus on human resources and focus on citizen-related governance from the following domains: media convergence, public and regulatory information policies, infrastructure management, real-time data mining and extraction, geographic information system, crowd computing, smart cities education, and social monitoring and control. In spite of the access to a great amount of data, we verify that the concept of intelligent city is referenced by a significant number of researches, but, in smaller number, works that present models of construction of a collective intelligence for the city. From this perspective, we identified the need to recognize technological education interventions for communication between individuals and the city. Because we believe that only through the implementation and management of techno-educommunication ecosystems will be able to promote a culture of participation.展开更多
AIM To investigate the role of tacrolimus intra-patient variability(IPV) in adult liver-transplant recipients.METHODS We retrospectively assessed tacrolimus variability in a cohort of liver-transplant recipients and a...AIM To investigate the role of tacrolimus intra-patient variability(IPV) in adult liver-transplant recipients.METHODS We retrospectively assessed tacrolimus variability in a cohort of liver-transplant recipients and analyzed its effect on the occurrence of graft rejection and de novo donor-specific antibodies(dn DSAs), as well as graft survival during the first 2 years posttransplantation. Between 02/08 and 06/2015, 116 patients that received tacrolimus plus mycophenolate mofetil(with or without steroids) were included. RESULTS Twenty-two patients(18.5%) experienced at least one acute-rejection episode(BPAR). Predictive factors for a BPAR were a tacrolimus IPV of > 35% [OR = 3.07 95%CI(1.14-8.24), P = 0.03] or > 40% [OR = 4.16(1.38-12.50), P = 0.01), and a tacrolimus trough level of < 5 ng/mL [OR=3.68(1.3-10.4), P =0.014]. Thirteen patients(11.2%) developed at least one dn DSA during the follow-up. Tacrolimus IPV [coded as a continuous variable: OR = 1.1, 95%CI(1.0-1.12), P = 0.006] of > 35% [OR = 4.83, 95%CI(1.39-16.72), P = 0.01] and > 40% [OR = 9.73, 95%CI(2.65-35.76), P = 0.001] were identified as predictors to detect dn DSAs. IPV did not impact on patient-or graft-survival rates during the follow-up. CONCLUSION Tacrolimus-IPV could be a useful tool to identify patients with a greater risk of graft rejection and of developing a de novo DSA after liver展开更多
This study examined the isolation and differentiation of dorsal root ganglion progenitor cells for therapeutic use in neurodegenerative diseases. Rat embryonic dorsal root ganglia progenitors were isolated and purifie...This study examined the isolation and differentiation of dorsal root ganglion progenitor cells for therapeutic use in neurodegenerative diseases. Rat embryonic dorsal root ganglia progenitors were isolated and purified using the differential adhesion method combined with cytosine arabinoside treatment. After culture in serum-free medium supplemented with B27, basic fibroblast growth factor and epidermal growth factor, these cells remained viable and survived for more than 18 months in vitro. Most cells differentiated to neurons that were immunoreactive for gamma-aminobutyric acid and choline acetyltransferase as detected by immunohistochemical staining. In addition, nerve growth factor and neurotrophic tyrosine kinase receptor expression were also observed in dorsal root ganglion progenitors and differentiated cells. K252a, an inhibitor that blocks nerve growth factor-induced signaling, inhibited cell survival, suggesting the possible existence of a nerve growth factor autocrine loop in these proliferating cells.展开更多
We assessed the effects of plantations of exotic trees (Pinus radiata, Eucalyptus globulus, and Populus nigra) on plant biodiversity in the temperate zone of the biodiversity hotspot of Central Chile. This region has ...We assessed the effects of plantations of exotic trees (Pinus radiata, Eucalyptus globulus, and Populus nigra) on plant biodiversity in the temperate zone of the biodiversity hotspot of Central Chile. This region has suffered from intense deforestation in favor of plantation establishment in the major part of the coastal area since the neoliberal turn in 1973. The approach presented aimed to analyze plant biodiversity on the α-, β- and γ-scale. Furthermore, a plantation evaluation index was applied in order to provide quantitative figures on management practices. Species richness is reduced by 50% below plantations. Diversity and evenness index values are also significantly reduced. Analyses on β-similarity indicate that plantations do not host species absent in adjacent native forests, and no additional habitat heterogeneity is gained. On the γ-scale, plantations lower the number of total species observed;especially of endemic species. The abundance of species considered as invasive is significantly higher and frequently, invasive plants dominate the understory. The evaluation index attests rather poor plantation management in Central Chile since plantations are grown as monocultures, natural elements and native species are lacking at specific sites and plantations are insufficiently connected to native plant formations at the landscape scale. Results give much concern since deforestation processes as observed in our study area is about to begin in Patagonia as well. If management practices from the temperate zone are adopted in Patagonia, a considerable decline in plant biodiversity has to be expected there.展开更多
Testicular sperm extraction is widely used in the treatment of male infertility in cases of non-obstructive azoospermia. Identifying spermatogenetic foci within the testes is critical for testicular sperm extraction. ...Testicular sperm extraction is widely used in the treatment of male infertility in cases of non-obstructive azoospermia. Identifying spermatogenetic foci within the testes is critical for testicular sperm extraction. Two-photon laser scanning microscopy (TPLSM) is an autofluorescence-based microscopy technique that allows observation at a cellular level in the depth of fresh living tissues and does not require any histological processing (fixation or staining). The wavelengths previously used have shown no phototoxicity on sperm. We used TPLSM to detect spermatogenetic foci in fresh mouse testicular parenchyma without disrupting the tunica albuginea. Fresh surgically retrieved testes were observed using TPLSM within 1 h after extraction. Contralateral testes for each animal were observed using standard histology. Using TPLSM we were able to observe and measure the diameter of seminiferous tubules through the tunica albuginea, similar to the histological control. Structures within epithelial tubules were also observed, although their nature has yet to be identified. TPLSM is a real-time microscopy technique that could detect spermatogenetic foci.展开更多
A collection of lactic acid bacteria isolates from fish viscera was studied and investigated regarding to their functional properties and safety aspects. From these, three isolates GM1, GM2 and GM3 were identified as ...A collection of lactic acid bacteria isolates from fish viscera was studied and investigated regarding to their functional properties and safety aspects. From these, three isolates GM1, GM2 and GM3 were identified as Enterococcus feacium species using molecular methods. Partial Amplified rDNA Restriction Analysis (partial ARDRA) with restriction enzyme HaeIII separated these isolates into distinctive group which suggest genotypic variability within enterococci strains isolated from fish viscera. The three strains GM1, GM2 and GM3 exhibited antimicrobial activity. Indeed strains have been shown to produce bacteriocins with inhibitory effect against food spoilage bacteria and pathogenic fish including Carnobacterium maltaromaticum. The molecular mass of bacteriocin, as calculated by tricine-SDS-PAGE, was found to be 4.5 kDa. All isolates were tested positive upon PCR amplification of enterocin A structural gene. Investigations of antibiotic resistance show that the isolates were mostly sensitive to several antibiotics (ampicillin, penicillin, tetracycline, gentamycine) and resistance to rifampicin. All isolates grow in esculin azide agar as a selective medium for enumeration of probiotic enterococci. This study suggests that our strains can be employed as probiotic or to improve the safety of food products.展开更多
The Amazon region is considered to be a high endemic area for Hepatitis B Virus (HBV) infections, Rondônia state having the highest prevalence. The aim of this study was to identify molecular genotypes and m...The Amazon region is considered to be a high endemic area for Hepatitis B Virus (HBV) infections, Rondônia state having the highest prevalence. The aim of this study was to identify molecular genotypes and mutations in the S gene region of HBV viral genomes from 20 patients using a DNA microarray. Results: Serological tests showed that 88% of patients were HBeAg negative, 82% had anti-HBe antibodies and 33% were co-infected with Hepatitis Delta Virus. Sixteen percent of the patients were considered cirrhotic, and 11% have been transfused. The microarray technique identified the genotypes A (4 patients), D (7 patients) and F (7 patients) in 18 samples. Mutations were detected in all 3 genotypes and, overall, A159G, which has been associated with a reduced antigenicity of the virus, was detected most frequently. In genotype A, G119E was the most frequently detected mutation followed by mutations A159G, F134Y, W172C, Y161F and T143S. A159G was detected in all genotype D and F samples followed by mutations T143S, Y161F, N131T, T114S and G119E in genotype D and mutations T143S, Y161F, N131T, T114S and G119E in genotype F. Conclusion: The analysis of mutations repartition among genotypes suggests that some of them are preferentially or exclusively associated with genotype A, D or F. This type of tool is adapted for clinical and therapy monitoring of patient as well as for molecular epidemiology research on HBV.展开更多
The study describes an analysis of the impact of plantation forestry on the taxonomic diversity of plants in south-central Chile. In this biodiversity hotspot, plantations of non-native species like Pinus radiata D. D...The study describes an analysis of the impact of plantation forestry on the taxonomic diversity of plants in south-central Chile. In this biodiversity hotspot, plantations of non-native species like Pinus radiata D. Don, Eucalyptus globules Labill. and Populus nigra L. have largely replaced native deciduous and sclerophyllous forests. The study compares taxonomic diversity of commercial plantations and native forests using taxonomic distinctness and diversity and the Simpson diversity index. Most of these indexes attest a considerably reduced taxonomic diversity to plantations. However, taxonomic distinctness values for P. radiata plantations seem to contradict other indexes at first glance. It is shown that the higher values of taxonomic distinctness of P. radiata plantations come from taxonomic dominance. Taxonomic dominance describes the fact that P. radiate—a member of the infradivison of Gymnospermae-bears only little taxonomic resemblance to other plants, which are Angiospermae. Thus, it strongly dominates the taxonomic distinctness index and the high taxonomic resemblance of other plant within its plots is neglected. Indexes are developed that identify such dominant species and adjust for taxonomic dominance in taxonomic diversity analyses. After this adjustment, all indexes provide a coherent image on taxonomic diversity. Plantation forestry produces a considerable decline of taxonomic diversity. Taxonomic diversity analysis provides valuable insights in biodiversity impacts and complements standard analyses.展开更多
文摘The Caatinga biome is an important ecosystem in the semi-arid region of Brazil.It has significantly degraded due to human activities and is currently a region undergoing desertification.Thus,monitoring the variation in the Caatinga biome has become essential for its sustainable development.However,traditional methods for estimating aboveground biomass(AGB)are time-consuming and destructive.Remote sensing,such as optical and radar imaging,can estimate and correlate with vegetation.Nevertheless,radar imaging is still a novelty to be applied in estimating the AGB of this biome,which is an area with little research.Therefore,this study aimed to use Sentinel-1 images to estimate the AGB of the Caatinga biome in Sergipe State(northeastern Brazil)and to verify its influencing factors.Nineteen sample plots(30 m×30 m)were selected,and the stems of individuals with a circumference at breast height(1.3 m above the ground)equal to or greater than 6.0 cm were measured,and the AGB through an allometric equation was estimated.The Sentinel-1 images from 3 different periods(green,intermediate,and dry periods)were used to consider the phenological conditions of the Caatinga biome.All the pre-processing and extraction of attributes(co-polarized VV(vertical transmit and vertical receive),cross-polarized VH(vertical transmit and horizontal receive),and band ratio VH/VV backscatter,radar vegetation index,dual polarization synthetic aperture radar(SAR)vegetation index(DPSVI),entropy(H),and alpha angle(α))were performed with Sentinel’s Application Platform.These attributes were used to estimate the AGB through simple and multiple linear regressions and evaluated by the coefficients of determination(R2),correlation(r),and root mean squared error(RMSE).The results showed that the attributes individually had little ability to estimate the AGB of the Caatinga biome in the three periods.Combined with multiple regression,we found that the intermediate period presented the equation with the best results among the observed and estimated variables(R^(2)=0.73;r=0.85;RMSE=8.33 Mg/hm^(2)),followed by the greenness period(R2=0.72;r=0.85;RMSE=8.40 Mg/hm^(2)).The attributes contributing to these equations were VH/VV,DPSVI,H,α,and co-polarized VV for the green period and cross-polarized VH for the intermediate period.The study showed that the Sentinel-1 images could be used to estimate the AGB of the Caatinga biome in the green and intermediate phenological periods since the SAR attributes highly correlated with the estimated variable(i.e.,AGB)through multiple linear equations.
基金Supported by Contrat Universite Paul Sabatier,Toulouse,France,ASUPS 2000(N.Vaysse)AOL DRC Hopitaux de Toulouse 2001,(L.Buscail)Region Midi-Pyrenees(L.Buscail)H.Laurell was supported by a grant from European Community Plan 99 ECC QLG3-CT-1999-0908(C.Susini)The Agilent 2100 Bioanalyzer and the phosphoimager(Molecular Dynamics,Sunnyvale,CA,USA)were at the Transcriptome Platform,Toulouse Genopole,and at the molecular biology platform at the Institute Louis Bugnard,IFR31,Toulouse,France,respectively
文摘AIM: To compare gene expression profiles of pancreatic adenocarcinoma tissue specimens, human pancreatic and colon adenocarcinoma and leukemia cell lines and normal pancreas samples in order to distinguish differentially expressed genes and to validate the differential expression of a subset of genes by quantitative real-time RT-PCR (RT-QPCR) in endoscopic ultrasound-guided fine needle aspiration (EUS-guided FNA) specimens.METHODS: Commercially dedicated cancer cDNA macroarrays (Atlas Human Cancer 1.2) containing 1176 genes were used. Different statistical approaches (hierarchical clustering, principal component analysis (PCA) and SAM) were used to analyze the expression data. RT-QPCR and immunohistochemical studies were used for validation of results.RESULTS: RT-QPCR validated the increased expression of LCN2 (lipocalin 2) and for the first time PLAT (tissue-type plasminogen activator or tPA) in malignant pancreas as compared with normal pancreas. Immunohistochemical analysis confirmed the increased expression of LCN2 protein localized in epithelial cells of ducts invaded by carcinoma. The analysis of PLAT and LCN2 transcripts in 12 samples obtained through EUS-guided FNA from patients with pancreatic adenocarcinoma showed significantly increased expression levels in comparison with those found in normal tissues, indicating that a sufficient amount of high quality RNA can be obtained with this technique.CONCLUSION: Expression profiling is a useful method to identify biomarkers and potential target genes. Molecular analysis of EUS-guided FNA samples in pancreatic cancer appears as a valuable strategy for the diagnosis of pancreatic adenocarcinomas.
基金INSERM,the Ministère de l'Education Nationale de la Recherche et de la Technologie,the Région Bretagne.No.2079
文摘AIM: TO study the implication of prokineticin 1 (PKI/EGVEGF) and prokineticin 2 (PK2/13v8) in hepatocellular carcinoma angiogenesis.METHODS: The gene induction of PK1/EG-VEGF and PK2/Bv8 was investigated in 10 normal, 28 fibrotic and 28 tumoral livers by using real time PCR. Their expression was compared to the expression of VEGF (an angiogenesis marker), vWF (an endothelial cell marker) and to CD68 (a monocyte/macrophage marker). Furthermore, the rnRNA levels of PK1/EG-VEGF, PK2/Bv8, prokineticin receptor 1 and 2 were evaluated by real time PCR in isolated liver cell populations. Finally, PK2/Bv8 protein was detected in normal liver paraffin sections and in isolated liver cells by immunohistochernistry and immunocytochemistry.RESULTS: PK2/Bv8 mRNA but not PK1/EG-VEGF was expressed in all types of normal liver samples examined. In the context of liver tumor development, we reported that PK2/13v8 correlates only with CD68 and showed a significant decrease in expression as the pathology evolves towards cancer. Whereas, VEGF and vWF mRNA were significantly upregulated in both fibrosis and HCC,as expected. In addition, out of all isolated liver cells examined, only Kupffer cells (liver resident macrophages) express significant levels of PK2/Bv8 and its receptors, prokineticin receptor 1 and 2.CONCLUSION: In normal liver PK2/Bv8 and its receptors were specifically expressed by Kupffer cells. PK2/Bv8 expression decreased as the liver evolves towards cancer and did not correlate with HCC angiogenesis.
基金Supported by funds from INSERM, the "Association pour la Recherche contre le Cancer" Grants, No. 3664, the "Region Midi Pyréeés". Audrey Ferrand was supported by the Ligue Nationale contre le cancer
文摘AIM: TO investigate in vivo, whether CCK2 receptors (CCK2R) regulate proteins known to play a crucial role in cell proliferation and cancer development and analyse in vitro the molecular mechanisms that lead to Src activation; in particular, to identify the domains within the CCK2R sequence that are implicated in this activation. METHODS: The expression and activation of Src and ERK were studied in vivo using immunofluorescence and western-blot techniques. We used pancreatic tissues derived from wild type or Elas-CCK2 mice that expressed the CCK2R in pancreatic acini, displayed an increased pancreatic growth and developed preneoplastic lesions. The pancreatic tumor cell line AR4-2J expressing the endogenous CCK2R or COS-7 cells transiently transfected with wild type or mutant CCK2R were used as in vitro models to study the mechanism of Src activation. Src activation was measured by in vitro kinase assays, ERK activation by western blot using antiphospho-ERK antibodies and the involvement of Src in gastrin-induced cell proliferation by MTT test. RESULTS: We showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, led to the activation of Src and the ERK pathway. Src was activated upstream of the ERK pathway by the CCK2R in pancreatic tumoral cells and contributed to the proliferative effects mediated by this receptor. In vitro results demonstrated that activation of the Src/ERK pathway by the CCK2R required the NPXXY motif, located within the CCK2R sequence at the end of the 7^th transmembrane domain, and suggested the putative role of Gq in this mechanism. CONCLUSION: Deregulation of the Src/ERK pathway by the CCK2R might represent an early step that contributes to cell proliferation, formation of preneoplastic lesions and pancreatic tumor development.
文摘Acute humoral rejection (AHR) is uncommon after ABO- compatible liver transplantation. Herein, we report two cases of AHR treated with plasmapheresis and rituximab in two ABO-compatible liver-transplant patients with preformed anti-human leukocyte antigen donor-specific antibodies. Patient 1 experienced a biopsy-proven AHR at day 10 post-transplant. She was treated by steroid pulses, and OKT3. Because of persisting signs of biopsy-proven AHR at day 26, she was treated by plasmapheresis and rituximab. Uver enzyme levels did not improve, and she died on day 41. Patient 2 experienced a biopsy-proven AHR on day 10 post-transplant. She was treated by steroid pulses, plasmapheresis, and rituximab. Liver enzymes returned to within normal range 18 d after diagnosis. Uver biopsies, at 3 and 9 mo post-transplant, showed complete resolution of AHR. We conclude that plasmapheresis should be started as soon as AHR is diagnosed, and be associated with a B-cell depleting agent. Rituximab may be considered as a first-line therapy.
基金Supported by a grant from l’Agence National de la Recherche sur le Sida (ANRS grant 2001/011)
文摘AIM: To evaluate the implication of substitutions in the hepatitis C virus (HCV) non-structural 5A (NS5A) protein in the resistance of HCV during mono-interferon (IFN) or combined IFN-ribavirin (IFN-R) therapy. Although NS5A has been reported to interact with the HCV RNA- dependent RNA polymerase, NS5B, as well as with many cellular proteins, the function of NS5A in the life cycle of HCV remains unclear. METHODS: HCV quasispecies were studied by clon- ing and sequencing of sequential isolates from patients infected by HCV genotype 1b. Patients were treated by IFN-α2b for 3 mo followed by IFN-α2b alone or com- bined IFN-R therapy for 9 additional months. Patients were categorized intro two groups based on their re- sponse to the treatments: 7 with sustained virological re- sponse (SVR) (quasispecies = 150) and 3 non-respond- ers (NR) to IFN-R (quasispecies = 106). RESULTS: Prior to treatment, SVR patients displayed a lower complexity of quasispecies than NR patients. Most patients had a decrease in the complexity of quasispe- cies during therapy. Analysis of amino acids substitu- tions showed that the degree of the complexity of the interferon sensitivity-determining region (ISDR) and the V3 domain of NS5A protein was able to discriminate thetwo groups of patients. Moreover, SVR patients displayed more variability in the NS5A region than NR patients. CONCLUSION: These results suggest that detailed mo- lecular analysis of the NS5A region may be important for understanding its function in IFN response during HCV 1b infection.
基金Supported by INSERMthe"Association pour la Recherche Contre le Cancer"Grants # 3664,# 4430the"Ligue Contre le Cancer"
文摘AIM: To investigate whether Src, JAK2 and phosphatidylinositol 3-kinase (PI3K) pathways are involved in the proliferation of human colonic tumour cells induced by glycine-extended gastrin (G-gly), the precursor of the mature amidated gastrin and to elucidate the molecular interaction between these three kinases in response to this peptide. METHODS: Using the human colonic tumour cell line HCT116 as a model, we first measured the activation of PI3K, p60-Src and JAK2 in response to G-gly by in vitro kinase assays. Then we investigated the involvement of these kinases in G-gly-induced cell proliferation by MTT test. RESULTS: G-gly stimulation induced p60-Src, JAK2 and PI3K activation in HCT116. The different pathways were involved in proliferation of human colon cancer cells induced by G-gly. Furthermore, we found that both Src and JAK2 were necessary to PI3K regulation by this peptide. However, we did not find any cross-talk between the two tyrosine kinases. CONCLUSION: Our results suggest that the p60-Src/ PI3K and JAK2/PI3K pathways act independently to mediate G-gly proliferative effect on human colonic tumour cells.
文摘Salmonella can invade non-phagocytic cells through its type Ⅲ secretion system (T3SS-1), which induces a Trigger entry process. This study showed that Salmonella enterica, subspecies enterica serovar Enteritidis can also invade cells via the Rck outer membrane protein. Rck was necessary and sufficient to enable non-invasive E. coli and Rckcoated beads to adhere to and invade different cells. Internalization analysis of latex beads coated with different Rck peptides showed that the peptide containing amino acids 140-150 promoted adhesion, whereas amino acids between 150 and 159 modulated invasion. Expression of dominant-negative derivatives and use of specific inhibitors demonstrated the crucial role of small GTPases Racl and Cdc42 in activating the Arp2/3 complex to trigger formation of actin-rich accumulation, leading to Rck-dependent internalization. Finally, scanning and transmission electron microscopy with Rck-coated beads and E. coli expressing Rck revealed microvillus-like extensions that formed a Zipper-like structure, engulfing the adherent beads and bacteria. Overall, our results provide new insights into the Salmonella T3SS-independent invasion mechanisms and strongly suggest that Rck induces a Zipper-like entry mechanism. Consequently, Salmonella seems to be the first bacterium found to be able to induce both Zipper and Trigger mechanisms to invade host cells.
文摘The objective of this research is to present a systematic analysis, in which we seek to identify the concepts of intelligent city, with the intention of recognizing the nature of this intelligence before the participation of the population. Therefore, the research associates concepts as smart cities, citizen participation and social web, identifying publications between 2012 and the first quarter of 2018 and questioning what the intelligent city is and how this concept is able to (re)organize the learning processes of the territory from the informal dynamics of the contemporary city. The qualitative analysis of the documents revealed an innumerable of definitions and related terms: smart, intelligent, ubiquitous, digital, knowledge, sustainable, crowd sourcing, innovative;structured in three types of approaches: technological focus, focus on human resources and focus on citizen-related governance from the following domains: media convergence, public and regulatory information policies, infrastructure management, real-time data mining and extraction, geographic information system, crowd computing, smart cities education, and social monitoring and control. In spite of the access to a great amount of data, we verify that the concept of intelligent city is referenced by a significant number of researches, but, in smaller number, works that present models of construction of a collective intelligence for the city. From this perspective, we identified the need to recognize technological education interventions for communication between individuals and the city. Because we believe that only through the implementation and management of techno-educommunication ecosystems will be able to promote a culture of participation.
文摘AIM To investigate the role of tacrolimus intra-patient variability(IPV) in adult liver-transplant recipients.METHODS We retrospectively assessed tacrolimus variability in a cohort of liver-transplant recipients and analyzed its effect on the occurrence of graft rejection and de novo donor-specific antibodies(dn DSAs), as well as graft survival during the first 2 years posttransplantation. Between 02/08 and 06/2015, 116 patients that received tacrolimus plus mycophenolate mofetil(with or without steroids) were included. RESULTS Twenty-two patients(18.5%) experienced at least one acute-rejection episode(BPAR). Predictive factors for a BPAR were a tacrolimus IPV of > 35% [OR = 3.07 95%CI(1.14-8.24), P = 0.03] or > 40% [OR = 4.16(1.38-12.50), P = 0.01), and a tacrolimus trough level of < 5 ng/mL [OR=3.68(1.3-10.4), P =0.014]. Thirteen patients(11.2%) developed at least one dn DSA during the follow-up. Tacrolimus IPV [coded as a continuous variable: OR = 1.1, 95%CI(1.0-1.12), P = 0.006] of > 35% [OR = 4.83, 95%CI(1.39-16.72), P = 0.01] and > 40% [OR = 9.73, 95%CI(2.65-35.76), P = 0.001] were identified as predictors to detect dn DSAs. IPV did not impact on patient-or graft-survival rates during the follow-up. CONCLUSION Tacrolimus-IPV could be a useful tool to identify patients with a greater risk of graft rejection and of developing a de novo DSA after liver
基金supported by the Rhone Alps Region of France, MIRA grants for the collaboration,the Agence National de la Recherche (ANR) RIB,the CNRS (International program for Scientific Cooperation, "PICS")
文摘This study examined the isolation and differentiation of dorsal root ganglion progenitor cells for therapeutic use in neurodegenerative diseases. Rat embryonic dorsal root ganglia progenitors were isolated and purified using the differential adhesion method combined with cytosine arabinoside treatment. After culture in serum-free medium supplemented with B27, basic fibroblast growth factor and epidermal growth factor, these cells remained viable and survived for more than 18 months in vitro. Most cells differentiated to neurons that were immunoreactive for gamma-aminobutyric acid and choline acetyltransferase as detected by immunohistochemical staining. In addition, nerve growth factor and neurotrophic tyrosine kinase receptor expression were also observed in dorsal root ganglion progenitors and differentiated cells. K252a, an inhibitor that blocks nerve growth factor-induced signaling, inhibited cell survival, suggesting the possible existence of a nerve growth factor autocrine loop in these proliferating cells.
文摘We assessed the effects of plantations of exotic trees (Pinus radiata, Eucalyptus globulus, and Populus nigra) on plant biodiversity in the temperate zone of the biodiversity hotspot of Central Chile. This region has suffered from intense deforestation in favor of plantation establishment in the major part of the coastal area since the neoliberal turn in 1973. The approach presented aimed to analyze plant biodiversity on the α-, β- and γ-scale. Furthermore, a plantation evaluation index was applied in order to provide quantitative figures on management practices. Species richness is reduced by 50% below plantations. Diversity and evenness index values are also significantly reduced. Analyses on β-similarity indicate that plantations do not host species absent in adjacent native forests, and no additional habitat heterogeneity is gained. On the γ-scale, plantations lower the number of total species observed;especially of endemic species. The abundance of species considered as invasive is significantly higher and frequently, invasive plants dominate the understory. The evaluation index attests rather poor plantation management in Central Chile since plantations are grown as monocultures, natural elements and native species are lacking at specific sites and plantations are insufficiently connected to native plant formations at the landscape scale. Results give much concern since deforestation processes as observed in our study area is about to begin in Patagonia as well. If management practices from the temperate zone are adopted in Patagonia, a considerable decline in plant biodiversity has to be expected there.
文摘Testicular sperm extraction is widely used in the treatment of male infertility in cases of non-obstructive azoospermia. Identifying spermatogenetic foci within the testes is critical for testicular sperm extraction. Two-photon laser scanning microscopy (TPLSM) is an autofluorescence-based microscopy technique that allows observation at a cellular level in the depth of fresh living tissues and does not require any histological processing (fixation or staining). The wavelengths previously used have shown no phototoxicity on sperm. We used TPLSM to detect spermatogenetic foci in fresh mouse testicular parenchyma without disrupting the tunica albuginea. Fresh surgically retrieved testes were observed using TPLSM within 1 h after extraction. Contralateral testes for each animal were observed using standard histology. Using TPLSM we were able to observe and measure the diameter of seminiferous tubules through the tunica albuginea, similar to the histological control. Structures within epithelial tubules were also observed, although their nature has yet to be identified. TPLSM is a real-time microscopy technique that could detect spermatogenetic foci.
文摘A collection of lactic acid bacteria isolates from fish viscera was studied and investigated regarding to their functional properties and safety aspects. From these, three isolates GM1, GM2 and GM3 were identified as Enterococcus feacium species using molecular methods. Partial Amplified rDNA Restriction Analysis (partial ARDRA) with restriction enzyme HaeIII separated these isolates into distinctive group which suggest genotypic variability within enterococci strains isolated from fish viscera. The three strains GM1, GM2 and GM3 exhibited antimicrobial activity. Indeed strains have been shown to produce bacteriocins with inhibitory effect against food spoilage bacteria and pathogenic fish including Carnobacterium maltaromaticum. The molecular mass of bacteriocin, as calculated by tricine-SDS-PAGE, was found to be 4.5 kDa. All isolates were tested positive upon PCR amplification of enterocin A structural gene. Investigations of antibiotic resistance show that the isolates were mostly sensitive to several antibiotics (ampicillin, penicillin, tetracycline, gentamycine) and resistance to rifampicin. All isolates grow in esculin azide agar as a selective medium for enumeration of probiotic enterococci. This study suggests that our strains can be employed as probiotic or to improve the safety of food products.
文摘The Amazon region is considered to be a high endemic area for Hepatitis B Virus (HBV) infections, Rondônia state having the highest prevalence. The aim of this study was to identify molecular genotypes and mutations in the S gene region of HBV viral genomes from 20 patients using a DNA microarray. Results: Serological tests showed that 88% of patients were HBeAg negative, 82% had anti-HBe antibodies and 33% were co-infected with Hepatitis Delta Virus. Sixteen percent of the patients were considered cirrhotic, and 11% have been transfused. The microarray technique identified the genotypes A (4 patients), D (7 patients) and F (7 patients) in 18 samples. Mutations were detected in all 3 genotypes and, overall, A159G, which has been associated with a reduced antigenicity of the virus, was detected most frequently. In genotype A, G119E was the most frequently detected mutation followed by mutations A159G, F134Y, W172C, Y161F and T143S. A159G was detected in all genotype D and F samples followed by mutations T143S, Y161F, N131T, T114S and G119E in genotype D and mutations T143S, Y161F, N131T, T114S and G119E in genotype F. Conclusion: The analysis of mutations repartition among genotypes suggests that some of them are preferentially or exclusively associated with genotype A, D or F. This type of tool is adapted for clinical and therapy monitoring of patient as well as for molecular epidemiology research on HBV.
文摘The study describes an analysis of the impact of plantation forestry on the taxonomic diversity of plants in south-central Chile. In this biodiversity hotspot, plantations of non-native species like Pinus radiata D. Don, Eucalyptus globules Labill. and Populus nigra L. have largely replaced native deciduous and sclerophyllous forests. The study compares taxonomic diversity of commercial plantations and native forests using taxonomic distinctness and diversity and the Simpson diversity index. Most of these indexes attest a considerably reduced taxonomic diversity to plantations. However, taxonomic distinctness values for P. radiata plantations seem to contradict other indexes at first glance. It is shown that the higher values of taxonomic distinctness of P. radiata plantations come from taxonomic dominance. Taxonomic dominance describes the fact that P. radiate—a member of the infradivison of Gymnospermae-bears only little taxonomic resemblance to other plants, which are Angiospermae. Thus, it strongly dominates the taxonomic distinctness index and the high taxonomic resemblance of other plant within its plots is neglected. Indexes are developed that identify such dominant species and adjust for taxonomic dominance in taxonomic diversity analyses. After this adjustment, all indexes provide a coherent image on taxonomic diversity. Plantation forestry produces a considerable decline of taxonomic diversity. Taxonomic diversity analysis provides valuable insights in biodiversity impacts and complements standard analyses.