Mesenchymal "stem" cells, or facilitators for the development of regenerative macrophages? Pericytes at the interface of wound healing.

Cultured mesenchymal stromal cells are among the most used cells in clinical trials. Currently, their potential benefits include provision of mature cell types through differentiation, and secretion of various types of paracrine signaling molecules. Even though research on these cells has spanned some decades now, surprisingly, their therapeutic potential has not been fully translated into clinical practice yet, which calls for further understanding of their intrinsic nature and modes of action. In this review, after discussing pieces of evidence that suggest that some perivascular cells may exhibit mesenchymal stem cell characteristics in vivo, we examine the possibility that subpopulations of perivascular and/or adventitial cells activated after tissue injury behave as MSCs and contribute to the resolution of tissue injury by providing cues for the development of regenerative macrophages at injured sites. Under this perspective, an important contribution of cultured MSCs (or their acellular products, such as extracellular vesicles) used in cell therapies would be to instigate the development of M2-like macrophages that support the tissue repair process.

Neuroprotective effects of the CTK 01512-2 toxin against neurotoxicity induced by 3-nitropropionic acid in rats.

The mitochondrial inhibitor 3-nitropropionic acid (3-NP) induces excitotoxicity. The authors hypothesized that CTK 01512-2, a recombinant peptide calcium channel N-type blocker, and the TRPA1 antagonist, could show neuroprotective effects. The male Wistar rats received 3-NP [25 mg/kg (i.p.) for 7 days], and a treatment of CTK 01512-2 was delivered intrathecally (i.t.), thrice a week. The neuroprotective effects were evaluated by [18F]FDG MicroPET analysis. The CTK 01512-2 toxin was able to reestablish similar glucose uptakes on the control animals. To detect the neurobehavioral effects from 3-NP, three protocols (6.25, 12.5, 18.75 mg/kg of 3-NP (i.p.), for 3, 4, and 6 days, respectively) were evaluated by performance tests (open field test, walk footprint, elevated plus-maze, Y-maze, and the object recognition test). Important disabilities in the gait of the rats were seen, as well as memory deficits, and anxious behavior in the animals that were treated with all 3-NP protocols. The dose of 18.75 mg/kg (for 3 days) showed the most pronounced behavioral effects and lethality, while the rats treated with 12.5 mg/kg (for 4 days) showed behavioral effects similar to the 6.25 mg/kg dose (for 6 days). The third protocol was then repeated and the rats were treated with the CTK 01512-2 toxin to be evaluated behaviorally again. The recombinant peptide prevented all of the gait-evaluated parameters that were induced by 3-NP at a 6.25 mg/kg dose, which displayed an improvement in the exploratory activities. Overall, these results have reinforced the positive effects of CTK 01512-2 against the behavioral changes that were induced by the mitochondrial inhibitor 3-NP.

In vivo treatment with a subacute low dose of 3-nitropropionic acid does not induce genotoxicity or mutagenicity in rats.

3-nitropropionic acid (3-NP) is a toxin that causes neural damage in the striatum and can lead to the development of Huntington's disease manifestations in animal models. Several studies have shown genotoxicity related to the 3-NP treatment. This study investigated potential genotoxicity and mutagenicity that was induced by a low dose (6.25 mg/kg i. p.) 3-NP subacute treatment (daily, over 6 days) in a rat model. The arterial blood and the frontal cortex were analyzed by the comet assay and the bone marrow by micronucleus. Surprisingly, the 3-NP subacute treatment with the low dose did not show genotoxic or mutagenic effects.

Prognostic utility of early plasma matrix metalloproteinases -2 and -9 concentrations after severe traumatic brain injury. / Valor prognóstico das concentrações precoces de metaloproteinases -2 e -9 após traumatismo craniencefálico grave.

OBJECTIVE: To determine whether the matrix metalloproteinases-2 and -9 plasma levels were associated with intensive care unit mortality in patients who suffered severe traumatic brain injury, despite the presence of extracerebral injuries. METHODS: This prospective cohort enrolled 39 male patients who suffered severe traumatic brain injury (Glasgow coma scale: 3 - 8 at hospital admission). The plasma matrix metalloproteinase -2 and matix metalloproteinase -9 levels were determined by ELISA at the time of intensive care unit admission. RESULTS: Severe traumatic brain injury was associated with a 46% intensive care unit mortality rate. Higher plasma matrix metalloproteinase -9 concentrations were associated with mortality: 147.94 ± 18.00ng/mL for survivors and 224.23 ± 23.86ng/mL for nonsurvivors (mean ± standard error of the mean, p = 0.022). In contrast, there was no significant association between matrix metalloproteinase -2 levels and intensive care unit mortality: 315.68 ± 22.90ng/mL for survivors and 336.55 ± 24.29ng/mL for nonsurvivors (p = 0.499). Additionally, there were no significant associations between matrix metalloproteinase -2 (p = 0.711) and matrix metalloproteinase -9 (p = 0.092) levels and the presence of associated lesions. CONCLUSION: Increased plasma matrix metalloproteinase -9 levels were associated with intensive care unit mortality following severe traumatic brain injury, regardless of the presence of extracerebral injuries. Conversely, in this same context, plasma matrix metalloproteinase -2 levels were not associated with short-term fatal outcome prediction.

OBJETIVO: Determinar se os níveis plasmáticos das metaloproteinases de matriz -2 e -9 tem associação com a mortalidade na unidade de terapia intensiva em pacientes com trauma craniencefálico grave, independentemente de lesões não cerebrais associadas. MÉTODOS: Esta coorte prospectiva incluiu 39 pacientes do sexo masculino com trauma craniencefálico grave (escore na escala de coma Glasgow na admissão hospitalar: 3 - 8). Os níveis plasmáticos das metaloproteinases -2 e -9 foram determinados por ELISA no momento da admissão na unidade de terapia intensiva. RESULTADOS: O trauma craniencefálico grave apresentou mortalidade de 46% na unidade de terapia intensiva. Concentrações mais elevadas de metaloproteinase -9 apresentaram associação com a mortalidade: 147,94 ± 18,00ng/mL para pacientes que sobreviveram e 224,23 ± 23,86ng/mL para os que não sobreviveram (média ± erro padrão, respectivamente; p = 0,022). Todavia, não houve associação significativa entre os níveis de metaloproteinase -2 e a mortalidade na unidade de terapia intensiva: 315,68 ± 22,90ng/mL para o grupo de sobreviventes e 336,55 ± 24,29ng/mL entre os pacientes que não sobreviveram (p = 0,499). Além disso, não se observaram associações significativas entre os níveis de metaloproteinase -2 (p = 0,711) ou metaloproteinase -9 (p = 0,092) e a presença de lesões não cerebrais associadas. CONCLUSÃO: Em vítimas de traumatismo craniencefálico grave, níveis elevados de metaloproteinase -9 tiveram valor preditivo para o desfecho fatal na unidade de terapia intensiva independentemente da presença de lesões não cerebrais associadas. Por outro lado, no mesmo cenário, os níveis plasmáticos de metaloproteinase -2 não apresentaram associação com a mortalidade na unidade de terapia intensiva.

Valor prognóstico das concentrações precoces de metaloproteinases -2 e -9 após traumatismo craniencefálico grave / Prognostic utility of early plasma matrix metalloproteinases -2 and -9 concentrations after severe traumatic brain injury

RESUMO Objetivo: Determinar se os níveis plasmáticos das metaloproteinases de matriz -2 e -9 tem associação com a mortalidade na unidade de terapia intensiva em pacientes com trauma craniencefálico grave, independentemente de lesões não cerebrais associadas. Métodos: Esta coorte prospectiva incluiu 39 pacientes do sexo masculino com trauma craniencefálico grave (escore na escala de coma Glasgow na admissão hospitalar: 3 - 8). Os níveis plasmáticos das metaloproteinases -2 e -9 foram determinados por ELISA no momento da admissão na unidade de terapia intensiva. Resultados: O trauma craniencefálico grave apresentou mortalidade de 46% na unidade de terapia intensiva. Concentrações mais elevadas de metaloproteinase -9 apresentaram associação com a mortalidade: 147,94 ± 18,00ng/mL para pacientes que sobreviveram e 224,23 ± 23,86ng/mL para os que não sobreviveram (média ± erro padrão, respectivamente; p = 0,022). Todavia, não houve associação significativa entre os níveis de metaloproteinase -2 e a mortalidade na unidade de terapia intensiva: 315,68 ± 22,90ng/mL para o grupo de sobreviventes e 336,55 ± 24,29ng/mL entre os pacientes que não sobreviveram (p = 0,499). Além disso, não se observaram associações significativas entre os níveis de metaloproteinase -2 (p = 0,711) ou metaloproteinase -9 (p = 0,092) e a presença de lesões não cerebrais associadas. Conclusão: Em vítimas de traumatismo craniencefálico grave, níveis elevados de metaloproteinase -9 tiveram valor preditivo para o desfecho fatal na unidade de terapia intensiva independentemente da presença de lesões não cerebrais associadas. Por outro lado, no mesmo cenário, os níveis plasmáticos de metaloproteinase -2 não apresentaram associação com a mortalidade na unidade de terapia intensiva

Abstract Objective: To determine whether the matrix metalloproteinases-2 and -9 plasma levels were associated with intensive care unit mortality in patients who suffered severe traumatic brain injury, despite the presence of extracerebral injuries. Methods: This prospective cohort enrolled 39 male patients who suffered severe traumatic brain injury (Glasgow coma scale: 3 - 8 at hospital admission). The plasma matrix metalloproteinase -2 and matix metalloproteinase -9 levels were determined by ELISA at the time of intensive care unit admission. Results: Severe traumatic brain injury was associated with a 46% intensive care unit mortality rate. Higher plasma matrix metalloproteinase -9 concentrations were associated with mortality: 147.94 ± 18.00ng/mL for survivors and 224.23 ± 23.86ng/mL for nonsurvivors (mean ± standard error of the mean, p = 0.022). In contrast, there was no significant association between matrix metalloproteinase -2 levels and intensive care unit mortality: 315.68 ± 22.90ng/mL for survivors and 336.55 ± 24.29ng/mL for nonsurvivors (p = 0.499). Additionally, there were no significant associations between matrix metalloproteinase -2 (p = 0.711) and matrix metalloproteinase -9 (p = 0.092) levels and the presence of associated lesions. Conclusion: Increased plasma matrix metalloproteinase -9 levels were associated with intensive care unit mortality following severe traumatic brain injury, regardless of the presence of extracerebral injuries. Conversely, in this same context, plasma matrix metalloproteinase -2 levels were not associated with short-term fatal outcome prediction.

Pain catastrophizing is associated with the Val66Met polymorphism of the brain-derived neurotrophic factor in fibromyalgia.

BACKGROUND: Fibromyalgia (FM) is a musculoskeletal chronic pain syndrome that impacts negatively patient's daily lives. Its pathogenesis is characterized by a complex relationship between biological and psychosocial factors not fully understood yet. Pain catastrophizing is associated with FM and is an important predictor of outcomes. This study aimed to answer two questions: (i) whether the allele and genotype frequencies of BDNF Val66Met (rs6265) polymorphism differs between FM patients and healthy controls (HC); and (ii) if the BDNF Val66Met polymorphism is a factor that predicts pain catastrophizing in FM. METHODS: In a cross-sectional design, 108 FM patients and 108 HC were included. FM patients responded to the Brazilian Portuguese version of the Pain Catastrophizing Scale (BP-PCS) to assess pain catastrophizing, as well as other validated tools for anxiety (The State-Trait Anxiety Inventory - STAI), depression (Beck Depression Inventory II - BDI-II) and functional aspects (Fibromyalgia Impact Questionnaire - FIQ; Central Sensitization Inventory validated and adapted for Brazilian population - CSI-BP; Pittsburgh Sleep Quality Index - PSQI; and Resilience Scale). All subjects were genotyped for the BDNF Val66Met polymorphism. RESULTS: Val allele was significantly more frequent in FM patients compared to the control group (p < 0.05). Also, FM patients with Val/Val genotype showed more pain catastrophizing thoughts, and this genotype was significantly associated with magnification and rumination dimensions of BP-PCS (p < 0.05). Furthermore, there were significant differences in levels of anxiety and symptoms of depression, years of education, and the functional situation between the FM and control groups. CONCLUSIONS: The findings show an association of BDNF Val66Met polymorphism with pain catastrophizing in FM, which opens new avenues to comprehend the interplay between molecular genetic characteristics and neuroplasticity mechanisms underpinning FM.

Pain catastrophizing is associated with the Val66Met polymorphism of the brain-derived neurotrophic factor in fibromyalgia

Abstract Background: Fibromyalgia (FM) is a musculoskeletal chronic pain syndrome that impacts negatively patient's daily lives. Its pathogenesis is characterized by a complex relationship between biological and psychosocial factors not fully understood yet. Pain catastrophizing is associated with FM and is an important predictor of outcomes. This study aimed to answer two questions: (i) whether the allele and genotype frequencies of BDNF Val66Met (rs6265) polymorphism differs between FM patients and healthy controls (HC); and (ii) if the BDNF Val66Met polymorphism is a factor that predicts pain catastrophizing in FM. Methods: In a cross-sectional design, 108 FM patients and 108 HC were included. FM patients responded to the Brazilian Portuguese version of the Pain Catastrophizing Scale (BP-PCS) to assess pain catastrophizing, as well as other validated tools for anxiety (The State-Trait Anxiety Inventory - STAI), depression (Beck Depression Inventory II -BDI-II) and functional aspects (Fibromyalgia Impact Questionnaire - FIQ; Central Sensitization Inventory validated and adapted for Brazilian population - CSI-BP; Pittsburgh Sleep Quality Index - PSQI; and Resilience Scale). All subjects were genotyped for the BDNF Val66Met polymorphism. Results: Val allele was significantly more frequent in FM patients compared to the control group (p < 0.05). Also, FM patients with Val/Val genotype showed more pain catastrophizing thoughts, and this genotype was significantly associated with magnification and rumination dimensions of BP-PCS (p < 0.05). Furthermore, there were significant differences in levels of anxiety and symptoms of depression, years of education, and the functional situation between the FM and control groups. Conclusions: The findings show an association of BDNF Val66Met polymorphism with pain catastrophizing in FM, which opens new avenues to comprehend the interplay between molecular genetic characteristics and neuroplasticity mechanisms underpinning FM.(AU)

Prognostic utility of circulating nucleic acids in acute brain injuries.

INTRODUCTION: Acute brain injuries represent major causes of morbidity and mortality worldwide. Nevertheless, therapeutic options are centered mainly on supportive care, and accurate prognosis prediction following traumatic brain injury (TBI) or stroke remains a challenge in clinical settings. Areas covered: Circulating DNA and RNA have shown potential as predictive molecules in acute brain injuries. In particular, plasma cell-free DNA (cfDNA) levels have been correlated to severity, mortality, and outcome after TBI and stroke. The real-time quantitative polymerase chain reaction (qPCR) is the most widely used technique for determination of cfDNA in brain injuries; however, to consider the use of cfDNA in emergency settings, a quicker and easier methodology for detection should be established. A recent study proposed detection of cfDNA applying a rapid fluorescent test that showed compatible results with qPCR. Expert commentary: As a promising perspective, detection of cfDNA levels using simple, rapid, and cheap methodology has potential to translate to clinic as a point-of-care marker, supporting the clinical decision-making in emergency care settings. Conversely, miRNA profiles may be used as signatures to determine the type and severity of injuries. Additionally, in the future, some miRNAs may constitute innovative neurorestorative therapies without the common hurdles associated with cell therapy.

Neurotrauma: The Crosstalk between Neurotrophins and Inflammation in the Acutely Injured Brain.

Traumatic brain injury (TBI) is a major cause of morbidity and mortality among young individuals worldwide. Understanding the pathophysiology of neurotrauma is crucial for the development of more effective therapeutic strategies. After the trauma occurs, immediate neurologic damage is produced by the traumatic forces; this primary injury triggers a secondary wave of biochemical cascades together with metabolic and cellular changes, called secondary neural injury. In the scenario of the acutely injured brain, the ongoing secondary injury results in ischemia and edema culminating in an uncontrollable increase in intracranial pressure. These areas of secondary injury progression, or areas of "traumatic penumbra", represent crucial targets for therapeutic interventions. Neurotrophins are a class of signaling molecules that promote survival and/or maintenance of neurons. They also stimulate axonal growth, synaptic plasticity, and neurotransmitter synthesis and release. Therefore, this review focuses on the role of neurotrophins in the acute post-injury response. Here, we discuss possible endogenous neuroprotective mechanisms of neurotrophins in the prevailing environment surrounding the injured areas, and highlight the crosstalk between neurotrophins and inflammation with focus on neurovascular unit cells, particularly pericytes. The perspective is that neurotrophins may represent promising targets for research on neuroprotective and neurorestorative processes in the short-term following TBI.

Plasma matrix metalloproteinase-9 levels predict intensive care unit mortality early after severe traumatic brain injury.

OBJECTIVES: Matrix metalloproteinase-9 (MMP-9) is an inducible metalloproteinase that can degrade the cerebrovascular matrix leading to disruption of the blood-brain barrier and exacerbation of oedema in neurotrauma. Therefore, our aim was to determine whether MMP-9 plasma levels were associated with intensive care unit (ICU) mortality after severe traumatic brain injury (TBI) despite the presence of extracerebral injuries. METHODS: This cohort enrolled 80 patients who suffered severe TBI (Glasgow Coma Scale: 3-8 at hospital admission). The plasma MMP-9 level was determined by enzyme-linked immunosorbent assay assay at ICU admission. RESULTS: Severe TBI was associated with a 32.5% ICU mortality rate. There was no association between the presence of extracerebral injuries (72.5% of the patients) and ICU mortality (P = 0.419). Higher plasma MMP-9 concentrations were associated with fatal outcome: 181.1 ± 16.0 ng/mL for survivors and 257.0 ± 23.2 ng/mL for nonsurvivors (mean ± S.E.M., P = 0.009). In contrast, there was no significant difference between MMP-9 levels and associated lesions: 220.8 ± 26.3 ng/mL for isolated TBI and 196.8 ± 15.8 ng/mL for patients with extracerebral injuries (P = 0.397). CONCLUSION: Increased plasma MMP-9 levels predicted short-term fatal outcome following severe TBI, regardless the presence of extracerebral injuries.

Plasma brain-derived neurotrophic factor levels after severe traumatic brain injury.

OBJECTIVE: Severe traumatic brain injury (TBI) is associated with a 30-70% mortality rate. Nevertheless, in clinical practice there are no effective biomarkers for the prediction of fatal outcome following severe TBI. Therefore, the aim was to determine whether brain-derived neurotrophic factor (BDNF) plasma levels are associated with intensive care unit (ICU) mortality in patients with severe TBI. METHODS: This prospective study enrolled 120 male patients who suffered severe TBI (Glasgow Coma Scale 3-8 at emergency room admission). The plasma BDNF level was determined at ICU admission (mean 6.4 hours after emergency room admission). RESULTS: Severe TBI was associated with a 35% mortality rate and 64% of the patients presented severe TBI with multi-trauma. The mean plasma BDNF concentration among the severe TBI victims was 704.2 ± 63.4 pg ml(-1) (±SEM). Nevertheless, there were no significant differences between BDNF levels in the survivor (700.2 ± 82.8 pg ml(-1)) or non-survivor (711.6 ± 97.4 pg ml(-1)) groups (p = 0.238) or in the isolated TBI (800.4 ± 117.4 pg ml(-1)) or TBI with multi-trauma groups (650.5 ± 73.9 pg ml(-1)) (p = 0.109). CONCLUSIONS: Plasma BDNF concentrations did not correlate with either short-term fatal outcome or type of injury following severe TBI.

Serum ferritin correlates with Glasgow coma scale scores and fatal outcome after severe traumatic brain injury.

OBJECTIVES: Severe traumatic brain injury (TBI) is associated with a 30-70% mortality rate. Nevertheless, in clinical practice there are no effective biomarkers for the prediction of fatal outcome following severe TBI. Therefore, the aim was to determine whether ferritin serum levels are associated with ICU mortality in patients with severe TBI. METHODS: This prospective study enrolled 69 male patients who suffered severe TBI [Glasgow Coma Scale (GCS) 3-8 at emergency room admission]. The serum ferritin protein level was determined at ICU admission (mean 5.6 ± 2.5 hours after emergency room admission). RESULTS: Severe TBI was associated with a 39% mortality rate. Higher serum ferritin concentrations were significantly associated with lower hospital admission GCS scores (p = 0.049). Further, there was a significant association between higher ferritin concentrations and fatal outcome (289.5 ± 27.1 µg L(-1) for survivors and 376.5 ± 31.5 µg L(-1) for non-survivors, respectively, mean ± SEM, p = 0.032). CONCLUSIONS: Increased serum ferritin levels were associated with lower hospital admission GCS scores and predicted short-term fatal outcome following severe TBI.

Prognostic value of circulating DNA levels in critically ill and trauma patients.

The number of studies investigating circulating nucleic acids as potential biomarkers has increased in recent years. The detection of such biomarkers is a minimally invasive alternative for the diagnosis and prognosis of various clinical conditions. The value of circulating DNA levels as a predictive biomarker has been demonstrated in patients suffering from numerous acute pathologies that have a high risk of intensive care needs and in-hospital deaths. The mechanism by which circulating DNA levels increase in patients with these conditions remains unclear. In this review, we focused on the potential use of this biomarker for prognosis prediction in critically ill and trauma patients. The literature review was performed by searching MedLine using PubMed in the English language.

Valor prognóstico dos níveis de DNA circulante em pacientes graves e em pacientes com trauma / Prognostic value of circulating DNA levels in critically ill and trauma patients

Nos últimos anos, o número de estudos que investigam os ácidos nucleicos circulantes como potenciais biomarcadores tem aumentado. A detecção desses biomarcadores é uma alternativa minimamente invasiva para o diagnóstico e o prognóstico de diversas condições clínicas. O valor dos níveis de DNA circulante como biomarcador preditivo foi demonstrado em pacientes com numerosas patologias agudas que apresentam riscos elevados de necessitar cuidados intensivos e de sofrer mortalidade hospitalar. Os mecanismos pelos quais os níveis de DNA circulante aumentam em pacientes com essas condições ainda são obscuros. Nesta revisão, focalizamos o potencial uso deste biomarcador para predição prognóstica em pacientes graves e pacientes com trauma. A revisão da literatura foi feita por meio de busca no MedLine utilizando o PubMed em inglês.

The number of studies investigating circulating nucleic acids as potential biomarkers has increased in recent years. The detection of such biomarkers is a minimally invasive alternative for the diagnosis and prognosis of various clinical conditions. The value of circulating DNA levels as a predictive biomarker has been demonstrated in patients suffering from numerous acute pathologies that have a high risk of intensive care needs and in-hospital deaths. The mechanism by which circulating DNA levels increase in patients with these conditions remains unclear. In this review, we focused on the potential use of this biomarker for prognosis prediction in critically ill and trauma patients. The literature review was performed by searching MedLine using PubMed in the English language.

Increased levels of interleukin-6, -8 and -10 are associated with fatal outcome following severe traumatic brain injury.

BACKGROUND: Despite the involvement of cytokine production in neurotrauma, there is still controversy regarding cytokines levels and clinical outcome following severe traumatic brain injury (TBI). OBJECTIVE: The present study was designed to investigate whether cytokine levels (of IL-1ß, IL-6, IL-8, IL-10, IL-12p70 and TNF-α) are associated with primary outcome (death or survival) after severe TBI. METHODS: This prospective study enrolled 24 male patients, victims of severe TBI. Venous blood samples were taken in the Intensive Care Unit (ICU) (study entry), 24 and 48 hours later. Plasma cytokine levels were assayed by flow cytometry. RESULTS: Severe TBI was associated with a 42% mortality rate. TBI patients had a significant increase in the levels of all cytokines measured, except for IL-1ß, compared to controls. Statistically significant increases in the IL-10, -8 and -6 levels were observed in the non-survivors TBI patients compared to the survivors sub-group measured in the first sample (study entry) and in the subsequent sample (24 hours later). There were no significant differences in IL-1ß, TNF-α and IL-12p70 levels between survivors and non-survivors in any time sampled. CONCLUSIONS: The findings indicate that increased IL-10, -8 and -6 levels may constitute an early predictor of unfavourable outcome in severe TBI patients.

Elevated cell-free plasma DNA level as an independent predictor of mortality in patients with severe traumatic brain injury.

Trauma is the leading cause of death in individuals less than 45 years old worldwide, and up to 50% of trauma fatalities are because of brain injury. Prediction of outcome is one of the major problems associated with severe traumatic brain injury (TBI), and research efforts have focused on the investigation of biomarkers with prognostic value after TBI. Therefore, our aim was to investigate whether cell-free DNA concentrations correlated to short-term primary outcome (survival or death) and Glasgow Coma Scale (GCS) scores after severe TBI. A total of 188 patients with severe TBI were enrolled in this prospective study; outcome variables comprised survival and neurological assessment using the GCS at intensive care unit (ICU) discharge. Control blood samples were obtained from 25 healthy volunteers. Peripheral venous blood was collected at admission to the ICU. Plasma DNA was measured using a real-time quantitative polymerase chain reaction (PCR) assay for the ß-globin gene. There was correlation between higher DNA levels and both fatal outcome and lower hospital admission GCS scores. Plasma DNA concentrations at the chosen cutoff point (≥171,381 kilogenomes-equivalents/L) predicted mortality with a specificity of 90% and a sensitivity of 43%. Logistic regression analysis showed that elevated plasma DNA levels were independently associated with death (p<0.001). In conclusion, high cell-free DNA concentration was a predictor of short-term mortality after severe TBI.

IL-6 polymorphism associated with fatal outcome in patients with severe traumatic brain injury.

OBJECTIVE: The aim of this study was to test whether a functional polymorphism (-174C/G) located in the promoter region of the interleukin-6 (IL-6) gene is associated with primary short-term outcome (death or Intensive Care Unit discharge) in patients with severe traumatic brain injury (TBI). METHODS: The study group consisted of 77 male patients who suffered severe TBI. The -174C/G IL-6 polymorphism was analysed by polymerase chain reaction (PCR) followed by restriction digestion. RESULTS: Severe TBI was associated with a 44% mortality rate. The GG genotype was significantly more frequent in the survivor group than in non-surviving patients (67% vs 41%; p =?0.038); similarly, the IL-6 -174G allele was more frequent in the survivor group than in non-surviving patients (81% vs 65%; p =0.031). CONCLUSION: The findings indicate that genetic variation regarding inflammatory response has significant impact on the short-term outcome for patients after acute severe TBI.

Glutamate promotes cell growth by EGFR signaling on U-87MG human glioblastoma cell line.

Accumulating evidences suggest that glutamate plays a key role in the proliferation and invasion of malignant glioblastoma (GBM) tumors. It has been shown that GBM cells release and exploit glutamate for proliferation and invasion through AMPA glutamate receptors. Additionally, amplification of the epidermal growth factor receptor (EGFR) gene occurs in 40-50% of GBM. Since, PI3K/Akt is considered one of the main intracellular pathways involved in EGFR activation, AKT functions could trigger EGFR signaling. Thus, we investigated whether EGFR-phospho-Akt pathway is involved on the glutamate inducing U-87MG human GBM cell line proliferation. For these purpose, we treated the U-87MG cell line with 5 to 200 mM of glutamate and assessed the number of viable cells by trypan blue dye exclusion test. An increase in cell number (50%) was found at 5 mM glutamate, while the addition of DNQX (500 microM), an antagonist of AMPA receptor, inhibited the effect of glutamate on the U87-MG cells proliferation. Also, at 5 mM glutamate we observed an increase on the EGFR and phospho-Akt contents evaluated by immunohistochemistry. Moreover, U-87MG cells treated with glutamate exhibited an increase about 2 times in the EGFR mRNA expression. While, in the presence of the anti-EGFR gefitinib (50 muM) or the PI3K inhibitor wortmannin (5 muM), the U-87MG proliferation was restored to control levels. Together, our data suggest that glutamate signaling mediated by AMPA receptor induces U-87MG human GBM cell line proliferation via EGFR-phospho-Akt pathway.

Ausência de associação entre polimorfismo do gene da interleucina-1 beta e o prognóstico de pacientes com traumatismo crânio-encefálico grave / Lack of association between interleukin-1 gene polymorphism and prognosis in severe traumatic brain injury patients

OBJETIVO: O traumatismo crânio-encefálico é a principal causa de óbito em indivíduos com idade entre 1 a 45 anos. O desfecho do traumatismo crânio-encefálico pode estar relacionado, além de fatores pré-morbidade e gravidade do dano, com fatores genéticos. Genes que podem ter relação com o resultado pós-trauma vêm sendo estudados, porém, ainda existem poucas informações sobre a associação entre polimorfismos genéticos e o desfecho do traumatismo crânio-encefálico. O gene da interleucina-1 beta (IL-1B) é um dos genes estudados, pois esta citocina encontra-se em níveis elevados após o traumatismo crânio-encefálico e pode afetar de forma negativa seu desfecho. O objetivo do presente estudo foi analisar o polimorfismo -31C/T, localizado na região promotora do gene IL-1B, em pacientes com traumatismo crânio-encefálico grave visando correlacioná-lo com o desfecho primário precoce (alta do centro de terapia intensiva ou morte). MÉTODOS: Foram estudados 69 pacientes internados por traumatismo crânio-encefálico grave em três hospitais de Porto Alegre e região metropolitana. O polimorfismo foi analisado através da reação em cadeia da polimerase, seguida da digestão com enzima de restrição. RESULTADOS: O traumatismo crânio-encefálico grave foi associado a uma mortalidade de 45 por cento. Não foram observadas diferenças significativas nas frequências alélicas e genotípicas entre os grupos de pacientes divididos pelo desfecho do traumatismo crânio-encefálico. CONCLUSÃO: Nossos resultados sugerem que o polimorfismo -31C/T do gene IL-1B não tem impacto significativo no desfecho fatal dos pacientes com traumatismo crânio-encefálico grave.

OBJECTIVE: Traumatic brain injury is the major cause of death among individuals between 1-45 years-old. The outcome of traumatic brain injury may be related to brain susceptibility to the injury and genetic factors. Genes that may affect traumatic brain injury outcome are being investigated, however there is still few data concerning the association between genetic polymorphisms and traumatic brain injury outcome. The interleukin-1 beta gene (IL-1B) is one of the most studied genes, because levels of this cytokine are raised after traumatic brain injury and this can affect worsen the prognosis. The aim of this study was to test whether the -31C/T polymorphism, located at the promoter region of the IL-1B gene, is associated with primary short-term outcome (death or intensive care unit discharge) in severe traumatic brain injury patients. METHODS: Were studied 69 patients admitted with severe traumatic brain injury in three hospitals of the metropolitan region of Porto Alegre. The polymorphism was analyzed by polymerase chain reaction, followed by restriction digestion. RESULTS: Severe traumatic brain injury was associated with a 45 percent mortality rate. No significant differences were observed in the allele and genotype frequencies between patients stratified by traumatic brain injury outcome. CONCLUSION: Our findings suggest that -31C/T IL-1B gene polymorphism have no significant impact on the outcome of patients after acute severe traumatic brain injury.

Quantitative plasma DNA analysis in Parkinson's disease.

Parkinson disease (PD) is a neurodegenerative disease resulting from the loss of the dopaminergic neurons from the substantia nigra pars compacta (SNpc). It is characterized by bradykinesia, rigidity, resting tremor and/or postural instability. The diagnosis of PD is essentially clinical and there is no reliable biological marker to assess its progression. Recently, investigations have been performed on the potential use of circulating cell-free deoxyribonucleic acid (DNA) in the plasma for clinical diagnosis, prognosis and monitoring of human diseases. The aim of this work was to assess the role of free DNA as a biological marker of PD. Forty-two patients with PD (19 men, 23 women) and 20 healthy (7 men, 13 women) subjects were enrolled in this study. Mean +/- SD plasma DNA concentration in PD patients and control subjects were, respectively, 16,487 +/- 16,378 (range: 100-62,034) kilogenomes-equivalents/L and 37,975 +/- 17,832 (range: 15,143-78,783) kilogenomes-equivalents/L. There was a significant difference between control and PD groups (p < 0.001). There was no correlation between plasma DNA levels and demographic or clinical parameters in PD patients. Free DNA does not seem to be a reliable marker of PD progression. Further research is warranted to confirm the present results to have some value as biomarkers in other neurodegenerative diseases.