Physical and thermal processing of Waste Printed Circuit Boards aiming for the recovery of gold and copper.

The recovery of electronic waste to obtain secondary raw materials is a subject of high relevance in the context of circular economy. Accordingly, the present work relies on the evaluation of mining separation/concentration techniques (comminution, size screening, magnetic separation and gravity concentration) alone as well as combined with thermal pre-treatment to recover gold and copper from Waste Printed Circuit Boards. For that purpose, Waste Printed Circuit Boards were subjected to physical processing (comminution, size screening in 6 classes from <0.425 mm to > 6.70 mm, magnetic separation and gravity concentration) alone and combined with thermal treatment (200-500 °C), aiming the recovery of gold and copper. Mixed motherboards and graphic cards (Lot 1 and 3) and highly rich components (connectors separated from memory cards, Lot 2) were analyzed. Gold and copper concentrations were determined before and after treatment. Before treatment, concentrations from 0.01 to 0.6 % wt. and from 9 to 20 % wt. were found for gold and copper respectively. The highest concentrations were observed in the size fractions between 0.425 and 1.70 mm. The highest copper concentration was around 35 % wt. (class 0.425-0.85 mm) and when analyzing memory card connectors alone, gold concentrations reached almost 2% in the same class, reflecting the interest of separating such components. The physical treatment alone was more effective for Lot 1/3, compared to Lot 2, allowing recoveries of 67 % wt. and 87 % wt. for gold and copper respectively, mostly due to differences in particles size and shape. The thermal treatment showed unperceptive influence on gold concentration but significant effect for copper concentration, mostly attributed to the size of the copper particles. Concentrations increased in a factor of around 10 when the thermal treatment was performed at 300 °C for the larger particles (1.70-6.70 mm); the best results were obtained at 400 °C for the other sizes, when the highest rate of thermal decomposition of the material occurred.

Effect of chromosome constitution variations on the expression of Turner phenotype.

Turner syndrome (TS) is a chronic disease related to haploinsufficiency of genes that are normally expressed in both X chromosomes in patients with female phenotype that is associated with a wide range of somatic malformations. We made detailed cytogenetic and clinical analysis of 65 patients with TS from the region of Recife, Brazil, to determine the effects of different chromosome constitutions on expression of the TS phenotype. Overall, patients with X-monosomy exhibited a tendency to have more severe phenotypes with higher morbidity, showing its importance in TS prognosis. Additionally, we found rare genetic and phenotypic abnormalities associated with this syndrome. To the best of our knowledge, this is the first case of 45,X,t(11;12)(q22;q22) described as a TS karyotype. Turner patients usually have normal intelligence; however, moderate to severe levels of mental retardation were found in 5 TS cases, which is considerate a very uncommon feature in this syndrome.

New cytogenetic aberrations found in a case of aggressive retinoblastoma.

We describe a case of retinoblastoma with an atypical presentation and previously unreported cytogenetic aberrations. A 19-month-old girl with left intraocular retinoblastoma was treated with enucleation and chemotherapy. The disease showed aggressive evolution within a short period between diagnosis and relapse. Eight months after diagnosis, a new large tumor was present in the orbit of the right eye, with diffuse bone pain, pancytopenia and diffuse infiltration into the bone marrow and the central nervous system. The child did not respond to treatment and died. Cytogenetic studies made with G-banding, FISH and SKY analysis showed chromosomal aberrations commonly associated with retinoblastoma, including del(13q), i(6p), +1, and monosomy 16, along with others that had not been reported previously, including dup(5q), dic(15;22) and add(14q). The new chromosomal aberrations may be related to the aggressiveness of the disease in this case.

An SOD rich melon extract Extramel prevents aortic lipids and liver steatosis in diet-induced model of atherosclerosis.

BACKGROUND AND AIMS: Oxidative stress has been involved in the early steps of atherosclerosis and previous studies on hypercholesterolemic hamsters have shown that non-enzymatic antioxidant could prevent fatty streak formation. Therefore, we investigated whether a melon juice extract (Extramel((R))) rich in superoxide dismutase (SOD) would prevent the development of early atherosclerosis. METHODS AND RESULTS: The effects of Extramel((R)) on plasma cholesterol, aortic fatty streak formation, hepatic steatosis, superoxide anion tissue production and NAD(P)H oxidase expression were studied in hamsters fed with an atherogenic diet (HF), receiving by gavage either water or Extramel((R)) at 0.7, 2.8 or 5.6mg/d. After 12 weeks of oral administration, Extramel((R)) lowered plasma cholesterol and non-HDL cholesterol and induced blood and liver SOD activities. It also strongly reduced the area of aortic fatty streak by 49-85%, cardiac (45%) and liver (67%) production of superoxide anion and liver p22(phox) subunit of NAD(P)H oxidase expression by 66%, and attenuated the development of hepatic steatosis. CONCLUSION: These findings support the view that chronic consumption of melon juice extract rich in SOD has potential beneficial effects with respect to the development of atherosclerosis and liver steatosis, emphasizing its use as potential dietary therapy.

Identification of a de novo inv dup(X)(pter--> q22) by multicolor banding in a girl with Turner syndrome.

We report on a 23-year-old girl with short stature, short and wide neck, low posterior hairline, hypogonadism, underdeveloped breasts, infantile uterus, ovaries not visualized, and primary amenorrhea. Cytogenetic G-banding analysis revealed a mosaic karyotype of 46,X,dup(X)(q22)[35]/45,X[15], confirming the clinical suspicion of Turner syndrome. Molecular cytogenetics using a multicolor banding probe set for the X-chromosome characterized an inverted dup(X). The karyotype of the patient was therefore interpreted as 46,X,inv dup(X) (pter --> q22::q22 --> pter). This patient had a mosaic Turner syndrome with a cell line comprising partial trisomy Xpter to Xq22 and partial monosomy Xq22 to Xqter.

Calcineurin Inhibitor Determination in Whole Blood With the RXL Dimension Analyzer: A Useful Tool for Immunosuppressive Drug Monitoring.

Routine monitoring of cyclosporine and tacrolimus levels is necessary to minimize adverse side effects and to ensure effective immunosuppression. The RXL Dimension apparatus conceived for ACMIA technologies is proposed to determine C0 and C2 cyclosporine levels and also tacrolimus levels in whole blood without any dilution or pretreatment using specific calibrators and Flex reagent cartridges (reagent stability: 72 hours for Neoral C0 and C2; 48 hours for tacrolimus). The assay ranges were between 25 to 500 ng/mL for C0; 350 to 2000 ng/mL for C2; and 1.2 to 30 ng/mL for tacrolimus. Within-run and between-day imprecision were <10% for cyclosporine. The coefficient of linearity was r(2) = .998 for C0, C2, and tacrolimus. Moreover, for cyclosporine and tacrolimus assays, the time for the first result was 20 minutes. Cyclosporine (C0, n = 152; C2, n = 54) and tacrolimus (n = 70) ACMIA assays were compared with enzyme-multiplied immunoassay technique (EMIT) cyclosporine and tacrolimus assays (V-Twin, Siemens ex-Dade Behring Laboratories) among 276 transplant patients: 119 kidney, 67 liver, 28 heart, and 62 bone marrow transplantations. Values obtained with the ACMIA assay were highly correlated with the EMIT assay for CsA C0 levels (ACMIA = 1.04 EMIT - 9.32; r(2) = .97); CsA C2 levels (ACMIA = 1.15 EMIT - 53.7; r(2) = .94); and tacrolimus levels (ACMIA = 0.93 EMIT - 0.16; r(2) = .93). In conclusion, the RXL Dimension analyzer is a useful tool for routine monitoring with a single method for C0 and C2 cyclosporine and tacrolimus level determinations in whole blood without any dilution or preanalytic treatment.

[Usefulness of brain natriuretic peptide and N-terminal pro-brain natriuretic peptide in the elderly]. / Intérêt du peptide natriurétique B et du N-terminal-proBNP chez le sujet âgé

B-type natriuretic peptide and N-terminal pro-brain natriuretic peptide are biomarkers secreted both in response to myocardial stretch and increased ventricular wall stress. They are now largely used in cardiovascular disease in adults, especially as diagnostic and prognostic tools. This article aims at synthesizing existing data concerning the usefulness of natriuretic peptides in elderly patients. With aging, co-morbidities such as renal failure are common and lead to an increase in circulating concentrations of natriuretic peptides. Nevertheless, their usefulness is relevant in geriatric practice. Their role has been well established in heart failure, not only in diagnosis, using two thresholds to decrease misclassification, but also in prognosis. In the future, natriuretic peptides could be used as prognosis tools in acute coronary syndrome, in cardiovascular risk assessment or in management of heart valve disease. However, further investigations in elderly people are needed before validating these indications in routine geriatric practice.

Effects of consuming sugars and alternative sweeteners during pregnancy on maternal and child health: evidence for a secondhand sugar effect.

Consumption of sugar and alternative low- or no-energy sweeteners has increased in recent decades. However, it is still uncertain how consumption of sugar and alternative sweeteners during pregnancy affects pregnancy outcomes and long-term offspring health. This review aims to collate the available evidence surrounding the consequences of sugar and alternative sweetener consumption during pregnancy, a so-called secondhand sugar effect. We found evidence that sugar consumption during pregnancy may contribute to increased gestational weight gain and the development of pregnancy complications, including gestational diabetes, preeclampsia and preterm birth. Further, we found a growing body of the animal and human evidence that maternal sugar intake during pregnancy may impact neonatal and childhood metabolism, taste perception and obesity risk. Emerging evidence also suggests that both maternal and paternal preconception sugar intakes are linked to offspring metabolic outcomes, perhaps via epigenetic alterations to the germline. While there have been fewer studies of the impacts of alternative sweetener consumption before and during pregnancy, there is some evidence to suggest effects on infant outcomes including preterm birth risk, increased infant body composition and offspring preference for sweet foods, although mechanisms are unclear. We conclude that preconception and gestational sugar and alternative sweetener consumption may negatively impact pregnancy outcomes and offspring health and that there is a need for further observational, mechanistic and intervention research in this area.

Brain Tumor-Enhancement Visualization and Morphometric Assessment: A Comparison of MPRAGE, SPACE, and VIBE MRI Techniques.

BACKGROUND AND PURPOSE: Postgadolinium MR imaging is crucial for brain tumor diagnosis and morphometric assessment. We compared brain tumor enhancement visualization and the "target" object morphometry obtained with the most commonly used 3D MR imaging technique, MPRAGE, with 2 other routinely available techniques: sampling perfection with application-optimized contrasts by using different flip angle evolutions (SPACE) and volumetric interpolated brain examination (VIBE). MATERIALS AND METHODS: Fifty-four contrast-enhancing tumors (38 gliomas and 16 metastases) were assessed using MPRAGE, VIBE, and SPACE techniques randomly acquired after gadolinium-based contrast agent administration on a 3T scanner. Enhancement conspicuity was assessed quantitatively by calculating the contrast rate and contrast-to-noise ratio, and qualitatively, by consensus visual comparative ratings. The total enhancing tumor volume and between-sequence discrepancy in the margin delineation were assessed on the corresponding 3D target objects contoured with a computer-assisted software for neuronavigation. The Wilcoxon signed rank and Pearson χ2 nonparametric tests were used to investigate between-sequence discrepancies in the contrast rate, contrast-to-noise ratio, visual conspicuity ratings, tumor volume, and margin delineation estimates. Differences were also tested for 1D (Response Evaluation Criteria in Solid Tumors) and 2D (Response Assessment in Neuro-Oncology) measurements. RESULTS: Compared with MPRAGE, both SPACE and VIBE obtained higher contrast rate, contrast-to-noise ratio, and visual conspicuity ratings in both gliomas and metastases (P range, <.001-.001). The between-sequence 3D target object margin discrepancy ranged between 3% and 19.9% of lesion tumor volume. Larger tumor volumes, 1D and 2D measurements were obtained with SPACE (P range, <.01-.007). CONCLUSIONS: Superior conspicuity for brain tumor enhancement can be achieved using SPACE and VIBE techniques, compared with MPRAGE. Discrepancies were also detected when assessing target object size and morphology, with SPACE providing more accurate estimates.

Matrix isolation FTIR spectroscopic and theoretical study of 3,3-dichloro-1,1,1-trifluoropropane (HCFC-243).

The molecular structure and infrared spectrum of the atmospheric pollutant 3,3-dichloro-1,1,1-trifluoropropane (HCFC-243) were characterized experimentally and theoretically. The theoretical calculations show the existence of two conformers, with the gauche (G) and trans (T) orientation around the HCCC dihedral angle. Conformer G was calculated to be more stable than form T by more than 10 kJ mol (-1). In consonance with the large predicted relative energy of conformer T, only the G form was identified spectroscopically in cryogenic argon (10 K) and xenon (20 K) matrices prepared from room-temperature equilibrium vapor of the compound. The observed infrared spectra of the matrix-isolated HCFC-243 were interpreted with the aid of high-level density functional theory calculations and normal coordinate analysis. For experimental identification of the weakest IR absorption bands, the spectrum of HCFC-243 in the neat solid state at 145 K was obtained. This spectrum also confirmed the sole presence of the G conformer in the sample. Natural bond orbital and atomic charge analyses were carried out for the two conformers to shed light on the most important intramolecular interactions in the two conformers, in particular those responsible for their relative stability.

Propranolol antagonizes the anti-inflammatory effect of alcohol and improves survival of infected intoxicated rabbits.

We studied the effects of alcohol and propranolol on the course of peritonitis in rabbits. Induction of sterile peritonitis with normal saline led to a 50% augmentation of granulocyte adherence in normal rabbits, and a mean cumulative granulocyte count of 27,000/mm(3) in peritoneal exudate by 8 h. Rabbits intoxicated with alcohol at the time of peritonitis induction maintained a granulocyte adherence below pretreatment values, and only delivered a cumulative mean of 12,000 granulocytes/mm(3) into the peritoneal fluid. When intoxicated rabbits received propranolol intravenously at the time of intoxication, adherence increased above preperitonitis levels, and stayed significantly above values for animals given alcohol alone. In addition, the defect in granulocyte delivery was prevented by propranolol, resulting in a mean cumulative granulocyte count in peritoneal fluid of 24,000/mm(3).When peritonitis was induced with live pneumococci instead of a sterile inflammatory stimulus, 14/18 normal animals survived the infection and were culture-negative when sacrificed at 2 wk. In contrast, 17/18 intoxicated animals died of the infection, in a mean of 2.8 days. 9 of 18 intoxicated animals who also received propranolol survived, and those who died lived a mean of 7.5 days. The survival rates and the time-to-death among the nonsurvivors given propranolol were both significantly greater than in the animals intoxicated without propranolol. Thus, propranolol prevents the granulocyte adherence and delivery defects induced by alcohol intoxication, and significantly improves survival from infection.

Molecular and cellular aspects of iron-induced hepatic cirrhosis in rodents.

Hepatic fibrosis and cirrhosis are common findings in humans with hemochromatosis. In this study we investigated the molecular pathways of iron-induced hepatic fibrosis and evaluated the anti-fibrogenic effect of vitamin E. Male gerbils were treated with iron-dextran and fed a standard diet or a alpha-tocopherol enriched diet (250 mg/Kg diet). In gerbils on the standard diet at 6 wk after dosing with iron, in situ hybridization analysis documented a dramatic increase of signal for collagen mRNA around iron foci onto liver fat storing cells (FSC), as identified by immunocytochemistry with desmin antibody. After 4 mo, micronodular cirrhosis developed in these animals, with nonparenchymal cells surrounding hepatocyte nodules and expressing high level of TGF beta mRNA. In this group, in vivo labeling with [3H]-thymidine showed a marked proliferation of nonparenchymal cells, including FSC. In iron-dosed gerbils on the vitamin E-enriched diet for 4 mo, in spite of a severe liver iron burden, a normal lobular architecture was found, with a dramatic decrease of collagen mRNA accumulation and collagen deposition. At the molecular level, a total suppression of nonparenchymal cell proliferation was appreciable, although expression of collagen and TGF beta mRNAs was still present into microscopic iron-filled nonparenchymal cell aggregates scattered throughout the hepatic lobule. In conclusion, our study shows that anti-oxidant treatment during experimental hepatic fibrosis arrests fibrogenesis and completely prevents iron induced hepatic cirrhosis mainly through inhibition of nonparenchymal cell proliferation induced by iron.

A theoretical prediction of stability in hydrogen-bonded complexes formed between oxirane and oxetane rings with HX (X=F and Cl).

The optimised geometries of heterocyclic hydrogen-bonded complexes, C2H4O...HX and C3H6O...HX, where X=F or Cl, were determined at DFT/B3LYP/6-311++G(d,p) computational level. Structural, electronic and vibrational properties of these complexes are used in order to compare the strained ring, which confer the great reactivity of these heterocyclic rings with monoprotic acids, forming a primary hydrogen bond. A secondary hydrogen bond between the hydrogen atoms of the CH2 groups and the halide species also takes place, thus causing a nonlinearity (characterized by the theta angle), in the primary hydrogen bond.

Lack of Association Between Elevated Urinary Levels of Interleukin-10 and Interferon Gamma With the Presence of Inflammation in Kidney Transplant Recipients.

INTRODUCTION: A lot of evidence has demonstrated the importance of different cytokines in acute renal rejection. Previous studies have examined the presence or absence of interleukin (IL)-10 in related immunopathologic rejection grafts as well as other interleukins. Studies in human transplantation show elevated levels of IL-10 and gamma interferon (INF-γ) in inflammation and rejection. OBJECTIVE: The objective of this study was to demonstrate the lack of association of elevated urinary levels of IL-10 and IFN in the presence of active inflammation. METHODS: An observational, descriptive, cross-sectional study conducted in transplant recipients at 12 months of follow-up after renal transplantation. In those who were held biopsy after renal transplantation at one year follow-up, or allograft dysfunction, we also measured IL-10 and INF-γ in the urine. The following were considered as variables: age, body mass index (BMI), gender, transplant type, creatinine, chronic kidney disease epidemiology collaboration equation, (CKD-EPI), modification of diet in renal disease study equation (MDRD), Banff classification, and levels of IL-10 and INF-γ. Statistical analysis was performed calculating a sample size of 25 patients, with an alpha bias of 0.05%, yielding measures of central tendency and determining no association between levels of IL-10 and INF-γ with the presence of rejection using SPSS 21.0 program. RESULTS: A total of 50 patients, 34 (68%) males, 16 (32%) females, with an average 31.7 ± 9.9 years, weight of 64.91 ± 13.84 kg, size 1.60 ± 0.10 m and 24.97 ± 4.07 BMI were included,39 (78%) living donor and 11 (22%) cadaveric. Twenty-six (52%) showed inflammation in the biopsy and 24 (48%) showed none. Mean creatinine was 1.81 ± 1.5, and the estimated glomerular filtration rate (eGFR) was 55.27 ± 22.46, 65.76 ± 26.7. (MDRD and CKD-EPI, respectively). No statistical difference was found in the levels of IL-10 and IFN-γ using analysis of variance. (ANOVA; P = .467 and P = .063, respectively) Based on Banff, the inflammation on biopsy score was 2.78 ± 2.84. There was statistical significance (P < .05) with respect to the Cr and eGFR by different equations. There were no significant interactions between cytokine levels and more than 1 factor. (as indicated by P < .2). DISCUSSION AND CONCLUSIONS: No significant differences were observed in the level of interleukins in patients with and without inflammation, denoting an adequate immunosuppression in most of these patients. Determination of inflammatory cytokines in urine could be used as a determinant of a good immunosuppression status, rather than as an early marker of rejection.

Transmembrane potential of liver mitochondria from hexachlorobenzene-and iron-treated rats.

The respiratory parameters and the membrane potential of liver mitochondria from rats treated with either hexachlorobenzene, iron or hexachlorobenzene plus iron, to induce experimental porphyria, have been studied. Partial uncoupling of oxidative phosphorylation has been observed in mitochondria from hexachlorobenzene- and hexachlorobenzene plus iron-treated rats. Direct evidence has been presented that this uncoupling is due to the action of pentachlorophenol endogenously formed by metabolism of hexachlorobenzene. No irreversible damage of mitochondria membrane has been revealed under both these conditions. Normal oxidative phosphorylation has been found in mitochondria from rats treated with iron alone. In contrast, they presented an anomalous membrane potential, fully restored by oligomycin. A possible involvement of lipid peroxidation process, induced by iron, in causing these abnormalities has been suggested.

The effect of ferric iron complex on isolated rat liver mitochondria. I. Respiratory and electrochemical responses.

Addition of iron(III)-gluconate complex to isolated rat liver mitochondria resulted in an increased iron content of mitochondria. Iron was accumulated through a relatively fast process (maximal uptake in less than 2 min incubation) by an energy-independent mechanism. The in vitro iron overload of mitochondria was associated with enhancement in the oxygen consumption, which was due to the induction of lipoperoxidative processes catalyzed by iron. It was found that a concentration of iron as low as 0.1 mM elicits a consistent production of malondialdehyde in mitochondria. Concomitant with the induction of lipoperoxidation a progressive fall in the mitochondrial membrane potential was observed. The occurrence of energy-consuming processes as a consequence of iron addition, and particularly the enhancement of endogenous Ca2+ cycling across the membrane, was suggested as the cause of the membrane potential drop.

The effect of ferric iron complex on isolated rat liver mitochondria. II. Ion movements.

It has been found that addition of iron(III)-gluconate complex to rat liver mitochondria disturbed the mitochondrial Ca2+ transport. Indirect evidence when the changes in the membrane potential during the transport of Ca2+ were followed, as well as direct evidence, when the fluxes of Ca2+ were monitored by a Ca2+-selective electrode, indicated that this iron complex induced an efflux of Ca2+ from liver mitochondria. The mechanisms by which iron induced Ca2+ release appeared to be linked to the induction of lipoperoxidation of mitochondrial membrane. The mitochondrial membrane, however, did not become irreversibly damaged under these conditions, as indicated by its complete repolarization. It was also shown that the induction by iron of lipoperoxidation brought about an efflux of K+ from mitochondria.

Relationship between number of allergic sensitizations and immunity impairment: anything new in immunotherapy for allergies and vaccines planning?

To 106 14-56 year-old allergic people (30 monosensitized, 24 sensitized to 2 pollens, 52 polysensitized) we have evaluated the Global Immune Competence Status (GICS). That's a compound score, made of ten parameters, six regarding cell-mediated immunity (WBC/mmc, Gr/mmc, Ly/mmc, Ly CD3+/mmc, Ly CD4+/mmc, CD4/CD8 Ratio), four regarding nutritional status and humoral immunity (Tot. Protein mg/dl, Albumin mg/dl, Gammaglobulins mg/dl, IgG mg/dl). Each parameter is brought on a grid including 4 worth scores worsening from 4 to 1, related to different ranges of values; this way quickly leads to characterize type and grade of immune deficiency. So doing we found that in 30 monosensitized people 27 (90%) show a complete immune competence, while just 3 people (10%) have impaired GICS: in these 1 (3%) regards cell-mediated immunity, while 2 (7%) regard humoral immunity. In 24 patients sensitized to 2 allergenes, 18 (75%) showed complete immune competence, while 6 (25%) a GICS impairment regarding cell-mediated immunity. In leaving 52 polysensitized patients, 30 people showed complete immune competence (58%), while 20 (38%) showed a GICS impairment regarding cell-mediated immunity and 2 (4%) impaired humoral immunity. This work shows that the higher the number of sensitizations is, the stronger the cell-mediated immunity impairment in allergic people become.

The expression of cytokine mRNA in the cauda equina of Lewis rats with experimental allergic neuritis.

Autoreactive CD4+ T cells can transfer experimental allergic neuritis (EAN) to naive recipients. In order to further analyze the role of these T cells and their corresponding cytokines in EAN, we studied the expression of mRNA for IFN-gamma, IL-4, and IL-10 in the cauda equina of rats with EAN using a quantitative competitive reverse transcriptase PCR method. Nerves were studied on days 0 (pre-immunization), 10 (disease onset), 13 (clinical progression), 16 (disease peak), as well as 20, 24, and 34 post immunization (recovery). IFN-gamma messages increased at disease onset and peaked at day 13 p.i. IL-10 message remained at a very low level at disease onset and surged at day 16. Both messages were low in recovery stage. IL-4 message was undetectable at any time point. These data suggest a pro-inflammatory role of IFN-gamma and anti-inflammatory role of IL-10 in EAN lesions. It is also possible that a clonal switch from Th1 to Th2 occurs in EAN lesions during the disease course.

Flow cytometric analysis of infiltrating cells in the peripheral nerves in experimental allergic neuritis.

Experimental autoimmune neuritis (EAN) is an animal model that shares clinical, pathological and electrophysiological features with the human disease Guillain-Barré syndrome (GBS). In this study, we isolated and characterized by fluorescent activated cell sorter (FACS) phenotype of the inflammatory cells infiltrating cauda equina (CE) of Lewis rats at the active stage of the disease. We found that at this stage of EAN macrophages (Mphi) and alphabeta T cells were two major populations isolated from CE. We also found that among total cell population isolated from CE, gammadelta T and NK cells composed two small but distinct populations, while B cells were negligible. We characterized phenotype of alphabeta T cells in CE as CD45RC(+)CD8(+) (activated cytotoxic lymphocytes) and CD45RC(-)CD4(+) (memory Th cells). The phenotype of gammadelta T cells was found to be consisted of only CD45RC(+)CD8(+) cells. Both alphabeta and gammadelta T cells in CE expressed a higher level of CD25, CD44 and CD54 activation markers compared to the other tissues. Immunohistochemistry demonstrated that gammadelta T cells existed apart from the intense cellular infiltrate. This is the first report on the isolation and FACS analysis of CE-infiltrating cells, contributing a new and alternative approach to study the inflammatory lesions in EAN. We conclude that both alphabeta and gammadelta T cells have a unique activation/inflammatory phenotype required to traffic through and be retained in the peripheral nerves during EAN.