Plasma n-6 Fatty Acid Levels Are Associated With CD4 Cell Counts, Hospitalization, and Mortality in HIV-Infected Patients.

BACKGROUND: Fatty acids, including n-6 series, modulate immune function, but their effect on CD4 cell counts, death, or hospitalization in HIV-infected patients on antiretroviral therapy is unknown. METHODS: In a randomized trial for effects of multivitamins in HIV-infected patients in Uganda, we used gas chromatography to measure plasma n-6 fatty acids at baseline; determined CD4 counts at baseline, 3, 6, 12, and 18 months; and recorded hospitalization or death events. The associations of fatty acids with CD4 counts and events were analyzed using repeated-measures analysis of variance and Cox regression, respectively. RESULTS: Among 297 patients with fatty acids measurements, 16 patients died and 69 were hospitalized within 18 months. Except for linoleic acid, n-6 fatty acids levels were positively associated with CD4 counts at baseline but not during follow-up. In models that included all 5 major n-6 fatty acids, age; sex; body mass index; anemia status; use of antiretroviral therapy, multivitamin supplements, and alcohol; and the risk of death or hospitalization decreased significantly with an increase in linoleic acid and gamma-linolenic acid levels, whereas associations for dihomo-gamma-linolenic acid, arachidonic acid, and aolrenic acid were null. The hazard ratios (95% confidence intervals) per 1 SD increase in linoleic acid and gamma-linolenic acid were 0.73 (0.56-0.94) and 0.51 (0.36-0.72), respectively. Gamma-linolenic acid remained significant (hazard ratio = 0.51; 95% confidence interval: 0.35 to 0.68) after further adjustment for other plasma fatty acids. CONCLUSIONS: Lower levels of gamma-linolenic acid are associated with lower CD4 counts and an increased risk of death or hospitalization. These results suggest a potential for using n-6 fatty acids to improve outcomes from antiretroviral therapy.

Clinical significance of serological biomarkers and neuropsychological performances in patients with temporal lobe epilepsy.

BACKGROUND: Temporal lobe epilepsy (TLE) is a common form of focal epilepsy. Serum biomarkers to predict cognitive performance in TLE patients without psychiatric comorbidities and the link with gray matter (GM) atrophy have not been fully explored. METHODS: Thirty-four patients with TLE and 34 sex - and age-matched controls were enrolled for standardized cognitive tests, neuroimaging studies as well as measurements of serum levels of heat shock protein 70 (HSP70), S100ß protein (S100ßP), neuronal specific enolase (NSE), plasma nuclear and mitochondrial DNA levels. RESULTS: Compared with the controls, the patients with TLE had poorer cognitive performances and higher HSP70 and S100ßP levels (p < 0.01). The patients with higher frequencies of seizures had higher levels of HSP70, NSE and S100ßP (p < 0.01). Serum HSP70 level correlated positively with duration of epilepsy (σ = 0.413, p < 0.01), and inversely with memory scores in the late registration (σ = -0.276, p = 0.01) and early recall score (σ = -0.304, p = 0.007). Compared with the controls, gray matter atrophy in the hippocampal and parahippocampal areas, putamen, thalamus and supplementary motor areas were found in the patient group. The HSP70 levels showed an inverse correlation with hippocampal volume (R square = 0.22, p = 0.007) after controlling for the effect of age. CONCLUSIONS: Our results suggest that serum biomarkers were predictive of higher frequencies of seizures in the TLE group. HSP70 may be considered to be a stress biomarker in patients with TLE in that it correlated inversely with memory scores and hippocampal volume. In addition, the symmetric extratemporal atrophic patterns may be related to damage of neuronal networks and epileptogenesis in TLE.

Qualidade espermática e perfil plasmático de lipídeos em garanhões suplementados com óleo de arroz semirefinado com alto teor de gama-orizanol / Spermatic quality and plasmatic lipids in stallions supplemented with rice bran oil semirefined containing high level of gamma-oryzanol

O gama-orizanol é uma substância natural contida no óleo de arroz, que por suas características hipocolesterolêmicas e antioxidantes, têm sido objeto de estudo em humanos e, mais recentemente, em equinos. Foram utilizados seis garanhões, de várias raças, com idade de 10±5,4 anos e peso inicial de 472,67±90,48 kg. Objetivou-se verificar os efeitos da suplementação da dieta com óleo de arroz semi-refinado, contendo 1,1% de gama-orizanol, sobre os níveis de lipídeos plasmáticos (colesterol total, HDL-C, LDL-C, VLDL-C e triglicérides), de testosterona e da qualidade espermática. Os garanhões foram divididos em dois grupos, recebendo em cada refeição 150 mL de óleo de soja ou de arroz durante 60 dias. Realizaram-se colheitas amostrais de sangue e sêmen a cada 15 dias do período experimental. O delineamento experimental foi inteiramente casualizado, com medidas repetidas no tempo, e as médias comparadas pelo teste F, considerando-se o nível de 5% de significância. Os valores médios de testosterona, colesterol total, HDL-C, LDL-C, VLDL-C e triglicérides foram respectivamente 75,93 ng/dL; 92,73; 61,47; 26,99 e 4,28 mg/dL para o tratamento com óleo de soja; e de 62,13 ng/dL; 110,20; 66,73; 38,44 e 5,02 mg/dL para o tratamento com óleo de arroz. Em relação à qualidade espermática, não foi observada diferença (p<0,05) entre os tratamentos nas variáveis: volume, motilidade, concentração e defeitos. Podemos concluir que a suplementação supracitada não influencia a qualidade espermática nem as concentrações plasmáticas de testosterona, VLDL-C, HDL-C e triglicérides, porém, pode elevar as concentrações plasmáticas de colesterol total e de LDL-C.

Gamma-oryzanol is a natural substance contained in rice bran oil that has been studied for years in humans due to its hypocholeterolemic and antioxidant properties, and in recent years in horses. Using six stallions from different breeds in a randomized experimental design, weighing 472.67±90.48 kg and aging 10±5.4 years, this study was aimed to evaluate the effects of supplementation with rice bran oil containing 1.1% of gamma-oryzanol in plasmatic lipids levels (total cholesterol, HDL-C, LDL-C, VLDL-C and triglycerides), testosterone and sperm quality. Stallions were divided in two groups, one receiving 150 mL of soybean oil, and the other 150 mL of rice bran oil twice a day, during 60 days. Blood and sperm samples were collected every 15 days of the trial period. Data obtained was processed, and means compared using F test, at 5% of confidence level. Mean values of total cholesterol, HDL-C, LDL-C, VLDL-C, triglycerides and testosterone were respectively 92.73; 61.47; 26.99; 4.28 mg/dL and 75.93 ng/dL in the soybean oil supplemented group, and 110.20; 66.73; 38.44; 5.02 mg/dL and 62.13 ng/dL in the rice bran oil one. No difference among treatments (p<0,05) were observed in sperm quality parameters volume, motility, concentration and defects. Supplementation of stallions with rice bran oil containing 1.1% of gamma-oryzanol do not promote any change in sperm quality, testosterone, HDL-C, VLDL-C or triglycerides plasmatic levels, but an increase in total cholesterol and LDL-C.

The plasma fraction of stored erythrocytes augments pancreatic cancer metastasis in male versus female mice.

BACKGROUND: Males with pancreatic cancer have decreased survival compared with females. Interestingly, perioperative blood transfusions have been shown to reduce survival in patients with pancreatic adenocarcinoma. Recent evidence incriminates blood transfusions from female donors as a causative factor in acute lung injury. We therefore hypothesize that male mice with pancreatic cancer will have greater tumor progression than female mice in response to transfusion. METHODS: Mice previously inoculated with pancreatic cancer cells received an intravenous injection of acellular plasma collected from single donor erythrocytes from either male or female donors. Control mice received an equal volume of intravenous saline. Necropsy to determine metastasis was performed in female mice at 4 wk status post-transfusion. The male group necessitated sacrifice at 3 wk post-transfusion due to clinical deterioration. RESULTS: Male mice developed more metastatic events than female mice, and this was accentuated when receiving blood from female donors. Male mice experienced weight loss within 2 wk of tail vein injection, and three mice in the male transfused groups died secondary to malignancy. Female mice did not manifest substantial weight loss, and did not die in the study time period. CONCLUSION: Male mice, compared with female, had significantly more metastatic events following transfusion of plasma from stored erythrocytes in an immunocompetent murine model of pancreatic adenocarcinoma. Moreover, the adverse effect of transfusion was augmented with female donor blood. These data are consistent with clinical outcomes from centers of excellence in treating pancreatic cancer and warrant further investigation.

Basic studies on the clinical applications of platelet-rich plasma.

Platelets, which contain many growth factors such as platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta), are being used in clinical applications as platelet-rich plasma (PRP). Only a few studies, however, have been conducted on the growth factors present in PRP and on the clinical applications using the drug delivery system (DDS). For the purpose of clinical application, we first modified the PRP preparation method and assessed the amounts of growth factors contained in the human platelet concentrates. Furthermore, we assessed fibrin glue as a DDS of platelet concentrates. Platelet precipitations were made by twice centrifuging human whole blood. The precipitated platelet was resuspended to yield the platelet concentrates. The growth factor concentrations were measured. Fibrin glue sheets containing this platelet concentrate were implanted in rabbit pinna and samples were obtained for immunostaining (anti-PDGF antibody) to assess the use of PRP over time using the fibrin glue as the DDS. The mean concentration of growth factors present in the platelet concentrates was three times or greater than that of conventional PRP. Furthermore, the results indicated that when the platelet concentrate was used with fibrin glue as a carrier, the contents were released over a period of about 1 week. This raises the possibility that this system may be useful in clinical applications.

Effects of perioperative recombinant human IFN-gamma (rHuIFN-gamma) application in vivo on T cell response.

In spite of the well-known immunoregulatory effects of recombinant human interferon-gamma (rHuIFN-gamma), in vitro clinical trials in trauma patients remain inconclusive. In vitro studies have shown that IFN-gamma has an effect on lymphocyte responses in addition to immunomodulatory effects on the monocyte/macrophage system. To investigate the in vivo effect of rHuIFN-gamma perioperatively on lymphocyte behavior in surgical patients, we studied 46 anergic patients undergoing major surgery. Treated patients (T, n = 24) received 100 microg rHuIFN-gamma subcutaneously (s.c.), and control patients (C, n = 22) received a placebo on preoperative days -7, -5, and -3 in a controlled, double-blinded placebo trial. Whole blood cultures were stimulated with mitogen on perioperative days, and cytokines were investigated in the supernatants. Interleukin-2 receptor (IL-2R) levels were significantly elevated in the treatment arm during the postoperative period (p < 0.05). The postoperative enhancement of IL-4 in C was completely attenuated in T (p < 0.05). IL-2 levels were elevated perioperatively in T but not in C. No significant effect of rHuIFN-gamma could be demonstrated on IL-10 or lymphocyte proliferation in vitro. From this pilot study, we conclude that preoperative in vivo immunomodulation of lymphocyte function with rHuIFN-gamma in anergic patients is effective. It improves immunoreactivity, as shown by elevated IL-2R levels. Elevated IL-2 and suppressed IL-4 levels indicate a shift toward a Th1-driven lymphocyte response.

Defects in leukocyte-mediated initiation of lipid peroxidation in plasma as studied in myeloperoxidase-deficient subjects: systematic identification of multiple endogenous diffusible substrates for myeloperoxidase in plasma.

More than a decade ago it was demonstrated that neutrophil activation in plasma results in the time-dependent formation of lipid hydroperoxides through an unknown, ascorbate-sensitive pathway. It is now shown that the mechanism involves myeloperoxidase (MPO)-dependent use of multiple low-molecular-weight substrates in plasma, generating diffusible oxidant species. Addition of activated human neutrophils (from healthy subjects) to plasma (50%, vol/vol) resulted in the peroxidation of endogenous plasma lipids by catalase-, heme poison-, and ascorbate-sensitive pathways, as assessed by high-performance liquid chromatography (HPLC) with on-line electrospray ionization tandem mass spectrometric analysis of free and lipid-bound 9-HETE and 9-HODE. In marked contrast, neutrophils isolated from multiple subjects with MPO deficiency failed to initiate peroxidation of plasma lipids, but they did so after supplementation with isolated human MPO. MPO-dependent use of a low-molecular-weight substrate(s) in plasma for initiating lipid peroxidation was illustrated by demonstrating that the filtrate of plasma (10-kd MWt cutoff) could supply components required for low-density lipoprotein lipid peroxidation in the presence of MPO and H(2)O(2). Subsequent HPLC fractionation of plasma filtrate (10-kd MWt cutoff) by sequential column chromatography identified nitrite, tyrosine, and thiocyanate as major endogenous substrates and 17 beta-estradiol as a novel minor endogenous substrate in plasma for MPO in promoting peroxidation of plasma lipids. These results strongly suggest that the MPO-H(2)O(2) system of human leukocytes serves as a physiological mechanism for initiating lipid peroxidation in vivo.

Counting of residual WBCs in WBC-reduced blood components: a multicenter evaluation of a microvolume fluorimeter by the United Kingdom National Blood Service.

BACKGROUND: Implementation of universal WBC reduction of blood components means that automated analytical methods may be the only satisfactory way for production laboratories to meet increased testing requirements. STUDY DESIGN AND METHODS: A multicenter study on the performance of a microvolume fluorimeter (IMAGN 2000, Becton Dickinson) was undertaken on 519 RBC, 353 platelet, and 27 fresh plasma units. RESULTS: WBC counts for the RBC samples ranged from 0.02 to 6.94 x 10(6) per unit (mean, 0.57) as determined by FC and from 0.02 to 5.53 x 10(6) per unit (mean, 0.40) as determined by MVF with a mean FC bias of +0.15 x 10(6) WBCs per unit, and discrepancies outside the 95% limits of agreement were mainly associated with higher FC counts. The series of platelet samples showed means of 0.90 (range, 0.06-19.45) and 0.66 (range, 0.01-18.95) x 10(6) WBCs per unit for FC and MVF methods, respectively. FC and MVF results were in good agreement at low counts, although significant discrepancies were noted at higher counts. Overall, for the platelet units, there was a mean FC bias of +0.34 x 10(6) WBCs per unit. The intermethod agreement exceeded 99 percent for both types of blood component when the single (both UK and United States) decision point of 5.0 x 10(6) WBCs per unit was applied. The mean WBC counts for the 27 analyzed fresh plasma units were 61.8, 56.0, and 46.0 per microL by Nageotte hemocytometry, FC, and MVF, respectively. CONCLUSIONS: This evaluation found that the level of intersite consistency for FC was relatively poor compared to that for MVF. The results nevertheless validated the broad equivalence of FC and MVF results for the current Council of Europe and UK/US decision points of <1.0 and <5.0 x 10(6) WBCs per unit.

Investigations into the haemorheological significance of postprandial and fasting hypertriglyceridaemia.

Two study designs were conceived to evaluate the rheological significance of hypertriglyceridaemia. We first investigated the course of serum- (SV) and plasma viscosity (PV) and erythrocyte aggregation in serum (SEA) and plasma (PEA) of healthy normolipidaemic individuals over 4 h after a fatty rich meal, in native material and after removal of triglyceride rich lipoproteins by centrifugation. Secondly, blood from patients with untreated hypertriglyceridaemia was investigated under fasting conditions. PEA and SEA increased in parallel with postprandial triglycerides (+135 mg dl-1), but the effect on PEA was more pronounced (+0.8 abs% increase; 2 h after the meal) as compared to SEA (+0.4 abs% increase). PV and SV increased in parallel to the same extent (+0.05 mPas). In the triglyceride poor infranatant no significant changes occurred. In fasting plasma PEA and PV were significantly lower (1.1 abs% and PV 0.04 mPas respectively) in infranatant than in native plasma, while only small differences in triglyceride (mostly VLDL) were observed. This phenomenon was barely detectable in serum samples. We conclude that triglyceride rich lipoproteins have a profound influence on haemorheological parameters, and that fibrinogen in particular, potentiates the effect of large fasting VLDL on plasma viscosity and erythrocyte aggregation.