Ionizing radiation modulates human macrophages towards a pro-inflammatory phenotype preserving their pro-invasive and pro-angiogenic capacities

In order to improve the efficacy of conventional radiotherapy, attention has been paid to immune cells, which not only modulate cancer cell response to therapy but are also highly recruited to tumours after irradiation. Particularly, the effect of ionizing radiation on macrophages, using therapeutically relevant doses, is not well understood. To evaluate how radiotherapy affects macrophage behaviour and macrophage-mediated cancer cell activity, human monocyte derived-macrophages were subjected, for a week, to cumulative ionizing radiation doses, as used during cancer treatment (2Gy/fraction/day). Irradiated macrophages remained viable and metabolically active, despite DNA damage. NF-kappaB transcription activation and increased Bcl-xL expression evidenced the promotion of pro-survival activity. A significant increase of pro-inflammatory macrophage markers CD80, CD86 and HLA-DR, but not CCR7, TNF and IL1B was observed after 10Gy cumulative doses, while anti-inflammatory markers CD163, MRC1, VCAN and IL-10 expression decreased, suggesting the modulation towards a more proinflammatory phenotype. Moreover, ionizing radiation induced macrophage morphological alterations and increased their phagocytic rate, without affecting matrix metalloproteases (MMP)2 and MMP9 activity. Importantly, irradiated macrophages promoted cancer cell-invasion and cancer cell-induced angiogenesis. Our work highlights macrophage ability to sustain cancer cell activities as a major concern that needs to be addressed to improve radiotherapy efficacy

Avaliação da qualidade de vida com o instrumento SF-36 em lombalgia crônica

OBJETIVO: Avaliar a qualidade de vida através dos domínios do Instrumento SF-36 em portadores de lombalgia crônica inespecífica. MÉTODOS: Trinta portadores de lombalgia crônica inespecífica foram aleatorizados para três grupos (grupo Iso (Isostretching), grupo RPG (Reeducação Postural Global) e Iso+RPG), e avaliados quanto à dor e qualidade de vida antes e após as intervenções fisioterápicas e reavaliados novamente após 2 meses de acompanhamento. Após a aceitação do Termo de Consentimento Livre e Esclarecido os pacientes foram submetidos a procedimentos como: avaliação fisioterápica através do Instrumento de Avaliação da Coluna Vertebral, Escala Visual Analógica de Dor (EVA), Questionário de Qualidade de Vida através do Instrumento SF-36, antes da 1ª sessão e após três meses de tratamento, e reavaliados 2 meses após o tratamento. RESULTADOS: Mostraram que ambas as técnicas fisioterápicas diminuíram a dor (p<0,001), porém quando foram associadas as duas técnicas (Iso + RPG) a melhora da dor foi significativamente maior, e na avaliação após dois meses de acompanhamento o método de RPG foi mais eficaz. Quanto à avaliação da qualidade de vida, as técnicas fisioterápicas foram eficazes após as intervenções (P<0,001), porém o método do Iso foi mais eficaz quando os pacientes foram reavaliados no acompanhamento. CONCLUSÃO: As técnicas fisioterápicas utilizadas neste estudo foram eficazes para tratar a lombalgia crônica inespecífica apresentada pelos pacientes, pois diminuíram a algia que os mesmos apresentavam e melhoraram a qualidade de vida segundo os domínios do Instrumento SF-36. Nível de Evidência II, Ensaio Clínico Controlado e Randomizado. ____________________________________________________________________________________ ABSTRACTThe objective of this study was to evaluate the quality of life (QL) with the use of the SF-36 Questionnaire in patients with chronic nonspecific low back pain (CNLBP). Thirty patients with CNLBP were randomly assigned to one of three groups (Iso group (Isostretching), GPR group (Global Postural Reeducation), and the Iso+GPR group. Patients underwent physical therapy assessment with the use of the Vertebral Spine Assessment, the Visual Analog Scale of Pain (VASP), and the SF-36 life quality questionnaire before the first session (first assessment), after three months of treatment (second assessment) and reassessed two months after the final session in the follow-up (third assessment). The results indicated that both physical therapy techniques reduced pain (p<0.001); when the techniques (Iso+GPR) were combined, the reduction in pain was significantly greater; and, in the follow-up assessment, the GPR method was more efficient. As for the QL, physical therapy techniques were effective after the interventions (p<0.001), and the Iso method was more effective when patients were reassessed in the follow-up. We conclude that the physical therapy techniques used in this study were efficient to treat CNLBP in the patients since they reduced pain and increased QL according to the results of the SF-36 questionnaire. Level of Evidence II, Randomized Controlled Clinical Trial

Purinergic signalling and immune cells

This review article provides a historical perspective on the role of purinergic signalling in the regulation of various subsets of immune cells from early discoveries to current understanding. It is now recognised that adenosine 5'-triphosphate (ATP) and other nucleotides are released from cells following stress or injury. They can act on virtually all subsets of immune cells through a spectrum of P2X ligand-gated ion channels and G protein-coupled P2Y receptors. Furthermore, ATP is rapidly degraded into adenosine by ectonucleotidases such as CD39 and CD73, and adenosine exerts additional regulatory effects through its own receptors. The resulting effect ranges from stimulation to tolerance depending on the amount and time courses of nucleotides released, and the balance between ATP and adenosine. This review identifies the various receptors involved in the different subsets of immune cells and their effects on the function of these cells

Purinergic signalling links mechanical breath profile and alveolar mechanics with the pro-inflammatory innate immune response causing ventilation-induced lung injury

Severe pulmonary infection or vigorous cyclic deformation of the alveolar epithelial type I (AT I) cells by mechanical ventilation leads to massive extracellular ATP release. High levels of extracellular ATP saturate the ATP hydrolysis enzymes CD39 and CD73 resulting in persistent high ATP levels despite the conversion to adenosine. Above a certain level, extracellular ATP molecules act as danger-associated molecular patterns (DAMPs) and activate the pro-inflammatory response of the innate immunity through purinergic receptors on the surface of the immune cells. This results in lung tissue inflammation, capillary leakage, interstitial and alveolar oedema and lung injury reducing the production of surfactant by the damaged AT II cells and deactivating the surfactant function by the concomitant extravasated serum proteins through capillary leakage followed by a substantial increase in alveolar surface tension and alveolar collapse. The resulting inhomogeneous ventilation of the lungs is an important mechanism in the development of ventilation-induced lung injury. The high levels of extracellular ATP and the upregulation of ecto-enzymes and soluble enzymes that hydrolyse ATP to adenosine (CD39 and CD73) increase the extracellular adenosine levels that inhibit the innate and adaptive immune responses rendering the host susceptible to infection by invading microorganisms. Moreover, high levels of extracellular adenosine increase the expression, the production and the activation of pro-fibrotic proteins (such as TGF-β, α-SMA, etc.) followed by the establishment of lung fibrosis

Frame shift mutation, exon skipping, and a two-codon deletion caused by splice site mutations account for pyruvate kinase deficiency

Three novel splice site mutations and two novel missense mutations were identified by molecular analysis of pyruvate kinase (PK) deficiency associated with hereditary nonspherocytic hemolytic anemia. A Nepalese PK variant, PK Kowloon, was found to have a homozygous transversion at the 5'- splice site of the seventh intervening sequence (IVS) of the L-type PK gene (Ivs7[+1]gt → tt). Using a reverse transcription polymerase chain reaction (RT-PCR) assay, we showed that the R-type PK mRNA in the proband's reticulocytes included the seventh IVS between the seventh and eighth exon, introducing a stop codon 3 nucleotides downstream of the mutated site. Consequently, the translational product may lack 44% of the R-PK polypeptide. A transition at the last nucleotide of axon 9 H (1269GCG → GCA) was found in a Japanese PK variant, PK 'Kamata.' The mutation did not alter the amino acid sequence, but caused skipping of the ninth exonic sequence in the R-PK transcripts. As a result, the affected R-type PK lost 51 amino acid residues (373Met-423Ala del). A transversion at the splice acceptor site of the third IVS (Ivs 3[-2]ag → tg) was identified in PK 'Aomori.' The mutation resulted in aberrant splicing at a cryptic splice site within exon 4, causing deletion of two codons in the aberrant R-PK transcript (95 Gly-96 Pro → del). Both PK 'Kamata' and PK 'Aomori' had a missense mutation on the other allele, 1044AAG → AAT (348Lys → Asn) and 1075CGC → TGC (359Arg → Cys), respectively. Although both 348Lys and 359Arg were located in the sixth loop of A domain (β/}a barrel, which has been shown to contain the substrate and cation binding sites, the degree of anemia was much more severe in PK 'Kamata' than PK 'Aomori,' possibly because the 51 amino acid deletion of PK 'Kamata' but the 2 amino-acid deletion of PK 'Aomori' may abolish PK catalytic activity.link_to_subscribed_fulltex