Innate and adaptive immunity gene expression of human keratinocytes cultured of severe burn injury

PURPOSE:Evaluate the expression profile of genes related to Innate and Adaptive Immune System (IAIS) of human Primary Epidermal keratinocytes (hPEKP) of patients with severe burns.METHODS: After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific IAIS PCR Arrays plates (SA Biosciences).RESULTS: After the analysis of gene expression we found that 63% of these genes were differentially expressed, of which 77% were repressed and 23% were hyper-regulated. Among these, the following genes (fold increase or decrease): IL8 (41), IL6 (32), TNF (-92), HLA-E (-86), LYS (-74), CCR6 (- 73), CD86 (-41) and HLA-A (-35).CONCLUSIONS: This study contributes to the understanding of the molecular mechanisms underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNIFESP-EPM Department of SurgeryUNIFESP-EPMUNIFESPUniversidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Department of SurgeryUNIFESP, EPM, Department of SurgeryUNIFESP-EPMUNIFESP, EPM, Department of SurgeryFAPESP: 2011/12945-4FAPESP: 2013/10.905-0SciEL

Innate and adaptive immunity gene expression of human keratinocytes cultured of severe burn injury

PURPOSE:Evaluate the expression profile of genes related to Innate and Adaptive Immune System (IAIS) of human Primary Epidermal keratinocytes (hPEKP) of patients with severe burns.METHODS: After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific IAIS PCR Arrays plates (SA Biosciences).RESULTS: After the analysis of gene expression we found that 63% of these genes were differentially expressed, of which 77% were repressed and 23% were hyper-regulated. Among these, the following genes (fold increase or decrease): IL8 (41), IL6 (32), TNF (-92), HLA-E (-86), LYS (-74), CCR6 (- 73), CD86 (-41) and HLA-A (-35).CONCLUSIONS: This study contributes to the understanding of the molecular mechanisms underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNIFESP-EPM Department of SurgeryUNIFESP-EPMUNIFESPUniversidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Department of SurgeryUNIFESP, EPM, Department of SurgeryUNIFESP-EPMUNIFESP, EPM, Department of SurgeryFAPESP: 2011/12945-4FAPESP: 2013/10.905-0SciEL

Keratinocyte growth factor, interleukins (1 beta, 6, 8, 10, 12), and tumor necrosis factor alpha in culture medium of dermal fibroblast of burned patients

PURPOSE:To evaluate the level of cytokines and keratinocyte growth factor (KGF) or Fibroblast Growth Factor 7 (FGF-7) in the culture medium of cultured human dermal fibroblasts from patients with large burn in comparison to small burn.METHODS:Fibroblasts of 10 patients (four large burns, four small burns and two controls) were initiated by the enzymatic method using collagenase. Cytokines and KGF in the supernatant of the culture medium was measured by, respectively, flow cytometry using Cytometric Bead Array Human Inflammation kit (CBA, BD Biosciences, USA) and the enzyme immunoassay method using the Quantikine (r) Human KGF. The experiments were performed in triplicate.RESULTS:The expression of IL-12 protein in patients with large burns showed a tendency to increase. IL- 6, IL- 10, and IL- 1beta were observed no difference. For IL - 8, TNF - alpha and KGF was observed a significant difference between the expression in large and small burned patient.CONCLUSION:That IL-8, TNF-alpha and KGF showed higher expression in cultured fibroblasts of large burned patients.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNIFESP-EPM Department of SurgeryUNIFESP-EPMUNIFESP-EPM Department of GynecologyUniversidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Department of SurgeryUNIFESP, EPM, Department of SurgeryUNIFESP-EPMUNIFESP, EPM Department of GynecologyUNIFESP, EPM, Department of SurgeryFAPESP: 2011/12945-4FAPESP: 2011/23.985-7SciEL

Toll like receptors gene expression of human keratinocytes cultured of severe burn injury

PURPOSE: To evaluate the expression profile of genes related to Toll Like Receptors (TLR) pathways of human Primary Epidermal keratinocytes of patients with severe burns.METHODS: After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific TLR pathways PCR Arrays plates (SA Biosciences).RESULTS: After the analysis of gene expression we found that 21% of these genes were differentially expressed, of which 100% were repressed or hyporegulated. Among these, the following genes (fold decrease): HSPA1A (-58), HRAS (-36), MAP2K3 (-23), TOLLIP (-23), RELA (-18), FOS (-16), and TLR1 (-6.0).CONCLUSIONS: This study contributes to the understanding of the molecular mechanisms related to TLR pathways and underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNIFESP-EPM Department of SurgeryUNIFESP-EPMUNIFESPUniversidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Department of SurgeryUNIFESP, EPM, Department of SurgeryUNIFESP-EPMUNIFESP, EPM, Department of SurgeryFAPESP: 2011/12945-4FAPESP: 2013/10.905-0SciEL

The making of multilateralist germany implications for US-german bilateral relations

“Em Novembro de 1989, o mundo assistia extasiado à queda do muro de Berlim. O simbolismo do acontecimento não se perdeu à medida que alemães orientais e ocidentais quebravam a materialização em cimento do que Winston Churchill anos antes chamara a “Cortina de Ferro”. A queda do muro de Berlim e consequente processo de reunificação, acompanhado pelo fim das divisões da Guerra Fria que rodeavam a Europa, criou mudanças tectónicas na geopolítica global. O processo de reunificação alemão, acompanhado por um espírito internacional de cooperação e optimismo, pendendo para a euforia, forneceu um modelo inspirador de diplomacia positiva e popular autodeterminação trabalhando conjuntamente para um bem comum – era uma diplomacia por excelência. Diplomatas e políticos em Washington, Moscovo, Londres e Paris lutaram para manter os acontecimentos pacíficos na Europa central e todo o processo de reunificação foi marcado por um espírito notável de internacionalismo; (...) Esse espírito de colaboração transatlântica contrasta com a situação uma década e meia mais tarde. Desde do final de 2002, muito foi feito no cada vez maior distanciamento cultural e político entre EUA e Europa. Este distanciamento assenta firmemente onde política externa e valores culturais convergem. O historiador britânico Timothy Garton Ash chamou-lhe “a crise do ocidente, a mais profunda desde do fim da Guerra Fria”

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost