Perinatal inflammation contributes to neurodevelopmental diseases, and animal models have revealed that the inflammatory response within the central nervous system is age dependent. It remains unclear what intrinsic and/or extrinsic factors are responsible for this variation. Here, my aim was to discover the mechanisms responsible for the age-dependent changes in the inflammatory response of the brain by injecting interleukin-1 (IL-1β) into the brain of mice at postnatal day (P)7, P14, P21 or into adult mice. A "window of susceptibility" was found at P14, which was associated with marked neutrophil recruitment and blood-brain barrier (BBB) breakdown, in response to a low dose of IL-1β. Evaluation of cytokine, chemokine, and adhesion molecule mRNA transcripts failed to reveal any specific increases in basal or reactive expression following the injection of IL-1β at P14. The extrinsic hepatic acute phase response (APR) was evaluated, but, once again, there was no evidence that an altered APR might account for the enhanced inflammatory response at P14. Indeed, an inverse relationship between the magnitude of the leukocyte recruitment to the brain and the APR was discovered. Enhancement of the APR with intravenous IL-1β after injection of a low dose of IL-1β into the brain was found to reduce the number of neutrophils and BBB permeability in the brain. While no molecular changes seem to account for the presence of the "window of susceptibility", a population of Iba-1⁺ large, flattened and irregular perivascular cells was discovered within the P14 brain, that may contribute to the increased leukocyte recruitment at P14. Although variations in the brain inflammatory response with development were not fully account for, my results highlight the importance of the systemic inflammatory response on the outcome of acute brain injury and suggest that the systemic APR might be manipulated therapeutically to protect the brain in the perinatal period.

Abstract The effects of perioperative filgrastim (rhG-CSF) and surgery per se on the postoperative acute phase reaction were studied by assessing leukocyte functions, cytokine levels and tenascin-C (Tn-C) and procollagen propeptide (PINP, PIIINP) concentrations in different body fluid compartments in patients undergoing gastrointestinal surgery. Thirty consecutive patients were randomized to receive either filgrastim or placebo for five days, starting 12 hours before colorectal surgery. Filgrastim treatment led to marked neutrophilia with decreased neutrophil migration in peripheral blood but not in peritoneal fluid 48 hours postoperatively. Neutrophil phagocytosis and bacterial killing did not differ between the groups. Filgrastim caused increased postoperative expression of neutrophil CD11b/CD18 in blood but not in peritoneal fluid or wound fluid. CD11b/CD18 expression was higher in both wound fluid and peritoneal fluid than in blood in the placebo group. The expression of neutrophil CD62L was higher in blood than in peritoneal fluid or wound fluid in both groups. The serum concentration of interleukin (IL)-8 was lower in the filgrastim group 5 hours postoperatively. The concentrations of IL-1β, IL-6, transforming growth factor (TGF)-β and IL-10 did not differ between the groups. The cytokine levels were markedly higher locally in the wound and in the peritoneal cavity compared to circulating blood. No adverse events attributable to filgrastim were seen. Leukocyte counts, neutrophil and monocyte functions and the levels of IL-6, IL-8 and granulocyte colony-stimulating factor (G-CSF) were measured from 18 patients before and after colorectal surgery. Surgery caused an increase in neutrophil and monocyte counts along with lymphocytopenia. Neutrophil phagocytosis was decreased 4 and 24 hours postoperatively, but normalized after that. A distinct systemic cytokine response was seen postoperatively. In a study with 24 patients, Tn-C concentration increased in wound fluid during the first postoperative week after abdominal surgery. The Tn-C level was markedly higher in wound fluid than in serum.

Pneumonia and sepsis are distinct but linked public health concerns. Each condition is the leading cause of the other; however, the responses controlling the susceptibility between the two disease processes remain speculative. The acute phase response (APR) is an important component of the host immune response during pneumonia and sepsis, and primarily driven by the activation of hepatocyte transcription factors NF-κB RelA and STAT3. While the NF-κB pathway is essential for inflammation and hepatocyte function, its inactivation has been associated with hepatotoxicity. Liver injury is an independent risk factor for sepsis morbidity and mortality, suggesting that pathways promoting liver homeostasis may limit the systemic consequences of pneumonia. To identify conditions in which NF-κB RelA is required for liver resilience, we challenged mice lacking hepatocyte RelA (hepRelAΔ/Δ) and wildtype (WT) controls with E. coli, K. pneumoniae, S. pneumoniae, LPS, or αGalCer to induce pneumonia, sepsis, and/or NKT cell activation. Severe hepatotoxicity was observed in hepRelAΔ/Δ mice in all conditions examined in association with apoptosis, which could be prevented by neutralization of TNFα. Lastly, these changes were associated with remodeling of the hepatic transcriptome, likely reflecting both the cause and consequence of hepatoxicity. We have previously shown that activation of STAT3 in hepatocytes limits pneumonia susceptibility during endotoxemia, but the mechanisms whereby this liver APR provides protection are unknown. Iron sequestration is a defense mechanism against bacterial infections, which require iron for growth. Based on previous observations that alveolar lining fluid is favorable for bacteria in the absence of liver STAT3, we investigated whether liver APR limits pneumonia susceptibility during sepsis by withholding iron to prevent bacterial outgrowth. WT mice or mice lacking hepatocyte STAT3 (hepSTAT3Δ/Δ) mice were challenged with endotoxemia followed by E. coli pneumonia, or cecal ligation and puncture (CLP). Induction of mRNA encoding several essential iron-regulating factors was ablated in hepSTAT3Δ/Δ mice after endotoxemia and pneumonia, and post CLP. Additionally, liver STAT3 activation significantly remodeled the pulmonary transcriptome during endotoxemia, which potentially represents other protective mechanisms. Taken together, these results suggest that hepatic APR is an important immunological interface modulating pneumonia and sepsis interaction and susceptibility.

Orientador: Aureo Evangelista Santana / Resumo: A hemorragia pulmonar induzida por exercício (HPIE) caracteriza-se pela ruptura de capilares pulmonares e extravasamento de sangue nas vias aéreas. Uma das formas mais utilizadas para diagnóstico é visibilização de sangue na traqueia por meio de exame endoscópico das vias aéreas. A HPIE ocasiona inflamação no sistema respiratório e a resposta de fase aguda leva à síntese de proteínas de fase aguda, como a amiloide A sérica (AAS). Diante disso, objetivou-se com a realização deste estudo avaliar se a HPIE causa aumento da AAS em equinos após corrida. Exame endoscópico das vias aéreas e amostras de sangue foram colhidas de equinos da raça Puro-sangue inglês (PSI), após corrida no Jockey Club Brasileiro, na Gávea, Rio de Janeiro. As amostras de soro foram utilizadas para efetuar análises de AAS, por meio de ensaio imunoenzimático pela técnica de ELISA indireto (Enzyme Linked Immuno Sorbent Assay), utilizando kit comercial. Os dados obtidos foram avaliados por meio de análise de variância e verificou-se diferenças (p ≤ 0,05) em relação à variável presença ou ausência de HPIE e a concentração de AAS. Setenta e um equinos, sendo 43 machos e 28 fêmeas, da raça PSI, foram distribuídos em dois grupos: o primeiro grupo, grupo controle (GC) com 28 animais (39%); o segundo, grupo hemorragia pulmonar induzida por exercício (GH), com 43 animais (61%). Houve diferença entre os grupos (p<0,0001) em relação à presença e à ausência de HPIE. A concentração de AAS no sangue dos animais dos grupos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Exercise-induced pulmonar hemorrhage (EIPH) is characterized as the rupture of pulmonary capillaries and blood loss within the respiratory tract. One of the most important diagnostic means is the presence of fresh blood at the trachea during endoscopic examination of the respiratory tract. EIPH causes inflammation of the respiratory system and may cause an acute phase response leading to synthesis of acute phase proteins such as serum amiloyd A (SAA). Therefore the aim of this study was to evaluate if EIPH causes increase in SAA of horses after race. Endoscopic exam and blood samples were collected from Thoroughbred racehorses after a race at the Jockey Club Brasileiro, at Gávea, Rio de Janeiro. Serum samples were used to perform SAA analyses by immunosorbent assay with indirect ELISA technique (Enzyme Linked Immuno Sorbent Assay), using a commercial kit. The collected data was evaluated by ANOVA (p ≤ 0.05) for presence or absence of EIPH and to evaluate SAA data. Seventy one Thoroughbred race horses, 43 males and 28 females, were used in this study and were divided into two groups. The control group (GC) was enlisted with 28 horses (39%) and the EIPH positive (GH) with 43 horses (61%). There was a significant difference in between both groups (p<0.0001) related to EIPH positive and negative horses. There was no difference on the SAA values between groups GC e GH (p= 0.292), and the mean values and mean standard error (X±MSE) were 4.9 ±3.1 µg/mL e 7.5 ±3.0 µg/mL, respectively... (Complete abstract click electronic access below) / Doutor

Alpha-tocopherol, the most abundant form of vitamin E in man, is transported in the circulation by plasma lipoproteins. It plays important roles, not only in preventing lipid peroxidation, but also in modulating several cell functions such as cell signaling and gene expression. While chylomicrons transport dietary alpha-tocopherol after intestinal absorption, LDL and HDL are the major carriers of alpha-tocopherol in fasting plasma and largely contribute to its delivery to cells and tissues. Exchanges of alpha-tocopherol occur between plasma lipoproteins. In addition, alpha-tocopherol transfers have also been observed, in both directions, between plasma lipoproteins and artificial chylomicrons such as intravenous lipid emulsion particles used in parenteral nutrition. In acute conditions, intravenous supply of vitamin E via lipid emulsions, which bypasses the intestinal tract, may offer some advantages over oral administration to rapidly increase alpha-tocopherol plasma concentration. However, many questions remain unanswered regarding kinetics and factors facilitating vitamin E exchanges between lipid emulsions and plasma lipoproteins. The first part of this work aimed at characterizing alpha-tocopherol transfers between alpha-tocopherol rich emulsion particles and plasma lipoproteins as well as the potential for plasma proteins to modulate such transfers. An in vitro model of incubation was used in which emulsion triglyceride concentration was relatively low and lipoprotein levels comparable to those commonly found in the circulation. Results indicate a high capacity for LDL and HDL to acquire extra-amounts of alpha-tocopherol by rapid mass transfers from alpha-tocopherol-rich emulsion particles. Data further shows that, at a fixed alpha-tocopherol concentration provided by emulsion particles, the limiting factor for alpha-tocopherol enrichment is not the capacity of plasma lipoproteins to accommodate extra-amounts of alpha-tocopherol but the facilitating effect of plasma proteins on alpha-tocopherol transfer, the duration of the incubation and possibly the competition between different acceptor particles. Two lipid transfer proteins, PLTP and CETP, appear to largely mediate facilitation of alpha-tocopherol transfer; however, other plasma proteins may be involved. Data further shows that alpha-tocopherol enriched LDL and HDL can readily transfer newly acquired alpha-tocopherol to cells, without any regulation by plasma proteins. Short-term prophylactic vitamin E supplementation has been suggested to be beneficial in some patients in acute conditions who present reduced plasma vitamin E concentrations in association with important changes in plasma lipids and severe oxidative stress. However, it was not clear whether low plasma vitamin E concentration in critically ill patients is related to changes in the composition of plasma lipoproteins or to a decrease in the number of alpha-tocopherol carriers. In the second part of this work, two clinical studies were conducted to analyze changes of lipoprotein concentration and composition in relation to inflammatory reaction and oxidative stress in selected subgroups of critically ill patients, namely patients undergoing cardiac surgery with different procedures. Important changes in LDL and HDL lipid content were observed, some of which contrast with previous observations made in critically ill septic patients. The reduced plasma level of alpha-tocopherol measured after cardiac surgery is entirely due to a reduced number of circulating LDL and HDL particles. Data suggests that such reduced number in alpha-tocopherol carriers post-surgery may impede the delivery of alpha-tocopherol to cells in conditions of increased requirements due to oxidative stress. Avoidance of extracorporeal circulation during cardiac surgery does not reduce inflammation-related changes in plasma lipids but largely prevents oxidative stress. This data on changes occurring in plasma lipoproteins may help to better define strategies against pro-inflammatory changes or oxidative stress. If further studies would confirm a clinical benefit with evidence-based rationale, alpha-tocopherol enriched lipid emulsions may be used to guarantee a sufficient alpha-tocopherol supply in acute conditions associated with fewer alpha-tocopherol transporters and increased requirements due to high risk of oxidative tissue injury.

Inflammatory diseases such as hypertension are associated with high levels of oxidative stress. Characteristic of oxidative stress is the inflammatory acute phase protein response. Oxidative stress and its accompanied inflammation can be reduced via phase 2 enzyme induction. Broccoli sprouts, a rich source of phase 2 enzyme inducers such as isothiocyanates, can be incorporated into the diet to increase phase 2 enzymes. <p> The hypothesis of this study is that, the dietary intake of dried broccoli sprouts, by inducing liver phase 2 enzymes, will decrease oxidative stress and the acute phase response in the blood of spontaneously hypertensive stroke-prone rats. <p> Spontaneously hypertensive stroke-prone rats (SHRsp) and Sprague Dawley (SD) rats were placed either on a control diet of modified AIN-93G or an experimental diet of modified AIN-93G supplemented with dried broccoli sprouts. The following parameters were examined: 1. Isothiocyanate absorption (an increased level of dithiocarbamates is reflective of ITC absorption), 2. Oxidative stress (a reduction in oxidative stress is evidenced by an increase in plasma protein thiols and blood glutathione (GSH)), 3. Acute phase proteins (a decreased APR is reflected by an increase in plasma albumin and a decrease in ceruloplasmin), 4. Activity of phase 2 enzymes (increased phase 2 enzyme induction results in higher activities of liver quinone reductase (QR), glutathione-S-transferase (GST) and glutathione reductase (GR)). <p> My experimental results demonstrated that broccoli sprout feeding results in higher protein thiol levels in female SHRsp and higher blood GSH levels in males but no acute phase protein changes were observed in either male or female SHRsp. Broccoli sprout feeding caused higher QR and lower GST activities in female SHRsp but did not affect the activities of phase 2 enzymes in male SHRsp. The activities of GST and QR were higher in SD rats than in SHRsp. Levels of dithiocarbamates were higher in the broccoli fed group than in the control fed group. <p> The results from this study do not present a clear pattern to support the hypothesis that dietary intake of broccoli sprouts by inducing phase 2 enzymes will decrease parameters of oxidative stress and the acute phase response. <p> In conclusion, there is an interactive role played by animal gender and the induction of phase 2 enzymes by dried broccoli sprouts.

Inflammatory diseases such as hypertension are associated with high levels of oxidative stress. Characteristic of oxidative stress is the inflammatory acute phase protein response. Oxidative stress and its accompanied inflammation can be reduced via phase 2 enzyme induction. Broccoli sprouts, a rich source of phase 2 enzyme inducers such as isothiocyanates, can be incorporated into the diet to increase phase 2 enzymes. <p> The hypothesis of this study is that, the dietary intake of dried broccoli sprouts, by inducing liver phase 2 enzymes, will decrease oxidative stress and the acute phase response in the blood of spontaneously hypertensive stroke-prone rats. <p> Spontaneously hypertensive stroke-prone rats (SHRsp) and Sprague Dawley (SD) rats were placed either on a control diet of modified AIN-93G or an experimental diet of modified AIN-93G supplemented with dried broccoli sprouts. The following parameters were examined: 1. Isothiocyanate absorption (an increased level of dithiocarbamates is reflective of ITC absorption), 2. Oxidative stress (a reduction in oxidative stress is evidenced by an increase in plasma protein thiols and blood glutathione (GSH)), 3. Acute phase proteins (a decreased APR is reflected by an increase in plasma albumin and a decrease in ceruloplasmin), 4. Activity of phase 2 enzymes (increased phase 2 enzyme induction results in higher activities of liver quinone reductase (QR), glutathione-S-transferase (GST) and glutathione reductase (GR)). <p> My experimental results demonstrated that broccoli sprout feeding results in higher protein thiol levels in female SHRsp and higher blood GSH levels in males but no acute phase protein changes were observed in either male or female SHRsp. Broccoli sprout feeding caused higher QR and lower GST activities in female SHRsp but did not affect the activities of phase 2 enzymes in male SHRsp. The activities of GST and QR were higher in SD rats than in SHRsp. Levels of dithiocarbamates were higher in the broccoli fed group than in the control fed group. <p> The results from this study do not present a clear pattern to support the hypothesis that dietary intake of broccoli sprouts by inducing phase 2 enzymes will decrease parameters of oxidative stress and the acute phase response. <p> In conclusion, there is an interactive role played by animal gender and the induction of phase 2 enzymes by dried broccoli sprouts.

Thesis (Ph. D.)--Case Western Reserve University, 2008. / [School of Medicine] Department of Biochemistry. Includes bibliographical references.

INTRODUCTION Tuberculosis remains a major public health threat globally and the Human Immunodeficiency Virus (HIV) pandemic afflicting developing and developed countries has resulted in enormous increases in tuberculosis infections worldwide. Researchers have previously documented very low plasma vitamin C levels in patients with active pulmonary tuberculosis. This was attributed to a number of factors including: accelerated turnover of vitamin C, shifts in plasma concentrations, increased collagen formation and tissue repair and decreased vitamin C intake. Vitamin C appears to have a role in steroid-genesis and catecholamine synthesis. Decreased plasma vitamin C levels may therefore impact on the stress hormone response and acute phase reaction of patients with active tuberculosis. AIM The primary aims of the study were to measure plasma vitamin C levels, as well as stress hormone levels and acute phase reaction in patients with newly diagnosed active pulmonary tuberculosis and control patients (without tuberculosis), to determine if there was any relationship between vitamin C levels and the levels of these other variables. METHODS AND MATERIALS This was a prospective study of seventy one (71) consecutive patients admitted to Helen Joseph Hospital (between March and October 2002) with newly diagnosed active pulmonary tuberculosis and eighty nine (89) control patients with medical conditions other than tuberculosis. Demographic, clinical and laboratory data were captured and analyzed using SPSS 7.5 soft-ware. Continuous variables were analyzed using students t-test. Categorical data were analyzed by non parametric analysis and Pearsons linear regression model was used to determine the correlation between vitamin C and the other variables in the two groups. RESULTS There were no differences in race, gender, age, suburb of residence and occupational distributions in the study group with tuberculosis compared to the control group. There were more smokers and consumers of alcohol in the control group (54 and 62 patients respectively) than in the study group (28 and 31 patients respectively). The study patients had lower blood pressure (average 90/40 mmHg versus 100/60 mmHg of controls), higher mean pulse rate (101.87 ± 15.14 beats/minute versus 82.92 ± 8.88 beats/minute, p< 0.01), higher mean temperature (38.66 ± 0.67oC versus 37.14 ± 0.44oC, p< 0.01), and lower body mass index (18.29 ± 3.80 Kg/ M2 versus 23.20 ± 5.35 Kg/ M2, p< 0.01). Laboratory data comparing study group and controls also showed marked differences as follows: White cell count (WCC) 8.68×106 / L ± 5.44 versus 11.00×106 / L ± 4.94, p = 0.01; Haemoglobin 9.56gm / dl ± 1.93 versus 12.92gm / dl ± 2.34, p < 0.01 and platelet count 369.21× 106/L ± 190.71 versus 295.94×106 / L ± 94.64, p = 0.01. White cell vitamin C levels (normal range – 20-40 μg/108 leucocytes) were low in half of the patients in both groups (study patients mean 29.85 ± 28.70μg/108 leucocytes versus controls 31.39 ± 30.24μg/108 leucocytes, p = NS). Plasma vitamin C levels were reduced (normal range 10-20 mg/ml) in both groups but more so in the controls (mean 3.87 ± 2.82 mg/ml versus 4.81 ± 3.21 mg / ml in study patients, p= 0.053). Mean cortisol levels were slightly higher in the study patients (448.11 ± 197.41ηmol/L) than controls (392.70 ± 191.25ηmol/L, p = NS). Norepinephrine levels were slightly higher in the study patients than controls (study patients mean 2531.61 ± 2043.60 ρmol/L versus 2178.98 ± 1719.98 ρmol/L of controls, p = NS). Dopamine levels were higher in the study patients than in the controls (468.42 ± 377.57 ρmol/L in study patients versus 293.37 ± 355.84 ρmol/L in controls, p = 0.01). Epinephrine levels were higher in the controls (control patients mean 680.64 ± 743.78 ρmol/L versus 449.41 ± 380.04 ρmol/L of the study patients, p = 0.03). Ferritin levels were much higher in the study patients compared with controls (study patients mean 3005.87 ± 5023.26 μg/L versus 466.51 ± 1774.76 μg/L of the controls, p<0.01) as were CRP levels (125.91 ± 54.77 mg/L in the study patients versus 77.22 ± 81.17 mg/L in the controls, p=0.01). Mean urine cotinine levels were 16.42 ± 24.26μM/L for controls and 9.28 ± 11.59 μM / L for the study patients (p=0.027). Correlation studies did not show any significant differences between the different variables. There was an inverse correlation between CRP levels and urine cotinine levels in the control group (R squared=0.058 and p= 0.024). DISCUSSION There were no differences in the demographic profile of the two groups. Smoking and alcohol consumption were more common in the control group than in the study patients. Over 90 % of patients in both groups had low plasma vitamin C levels, while half of the patients in each group had low white cell vitamin C levels. The low levels of vitamin C could be due to some of the reasons given above or possibly due to the fact that generally there are low levels in Africans for reasons that are not apparent. The control group had increased mean urine cotinine levels suggesting a possible influence of cigarette smoking on vitamin C homeostasis in these patients. In both groups, the majority of patients had normal cortisol levels as well as normal to high catecholamine levels. Also, Ferritin and CRP levels were much higher in the study group than in the controls. The low levels of vitamin C did not, however, have any relationship with stress hormone levels and acute phase reactants. CONCLUSION This study has reaffirmed low plasma and white cell vitamin C levels in patients with new onset pulmonary tuberculosis but has also found low levels in control patients with diseases other than pulmonary tuberculosis. The study demonstrates adequate stress hormone responses in tuberculosis patients, which was not different from non- tuberculosis patients. Acute phase responses were found to be of higher magnitude in tuberculosis patients than in the controls. There were, however, no correlations between plasma vitamin C and stress hormones or acute phase reactants.

No description available.