Anethole and eugenol reduce in vitro and in vivo leukocyte migration induced by fMLP,LTB4, and carrageenan

The aim of this study was to investigate the effect of anethole (AN) and eugenol (EUG) on leukocyte migration using in vitro chemotaxis and in situ microcirculation assays. BALB/c mice were used for the in vitro chemotaxis assay, and Wistar rats for the in situ microcirculation assay. We evaluated (a) the in vitro leukocyte migration in response to chemotactic factors (formyl-methionyl-leucyl-phenylalanine [fMLP] and leukotriene B₄ [LTB₄]) and (b) the rolling, adhesion, and migration of leukocytes induced by an injection of carrageenan (100 µg/cavity) into the scrotum of the animal. In the in vitro chemotaxis assay, AN and EUG at doses of 1, 3, 9, and 27 µg/ml significantly inhibited leukocyte migration when stimulated by the chemotactic agents fMLP and LTB4. In the in situ microcirculation assay, AN at doses of 125 and 250 mg/kg and EUG at a dose of 250 mg/kg significantly decreased the number of leukocytes that rolled, adhered, and migrated to perivascular tissue. The results indicate that AN and EUG exert inhibitory effects on leukocyte migration, highlighting their possible use to diminish excessive leukocyte migration in the inflammatory process.     

Effects of LED phototherapy on relative wound contraction and reepithelialization during tissue repair in hypothyroid rats: morphometric and histological study

The aim of this study was to assess morphometrically and histologically, the effects of light-emitting diode (LED) (λ630?±?20 nm) phototherapy on reepithelialization and wound contraction during tissue repair in hypothyroid rats. Thyroid hormone deficiency has been associated with disorders of tissue repair. LED phototherapy has been studied using several healing models, but their usefulness in the improvement of hypothyroidism wound healing remains unknown. Under general anesthesia, a standard surgical wound (1 cm²) was produced on the dorsum of 48 male Wistar rats divided into four groups of 12 animals each: EC—control euthyroid, ED—euthyroid + LED, HC—control hypothyroid, and HD—Hypothyroid + LED. The irradiation started immediately after surgery and was repeated every other day for 7 and 14 days. Photographs of the wound were taken at the day of the surgical procedure and on days 8 and 15 after surgery, when animals’ deaths occurred. The specimens were removed, routinely processed, and stained with hematoxylin/eosin. Seven days after the surgery, it was possible to observe statistically significant reductions in the wound area of the irradiated euthyroid group, in comparison to hypothyroid group, irradiated and non-irradiated (ANOVA, p?<?0.05). The reepithelialization was significantly higher in the euthyroid and hypothyroid groups irradiated with LED than in the non-irradiated groups (Fisher’s test, p?<?0.05). No significant difference was found in the experimental period of 14 days among the groups. The hypothyroidism delayed wound healing and the LED phototherapy, at these specific parameters, improved the process of reepithelialization in the presence of hypothyroidism.     

Endophytic species of Colletotrichum associated with mango in northeastern Brazil

Endophytic species of Colletotrichum associated with Mangifera indica (mango) are poorly understood. In this study, Colletotrichum species were isolated from mango in Pernambuco State, Brazil. There were significant differences in isolation frequencies of Colletotrichum species among sites and plant tissues. Mature leaf blades were colonized by most Colletotrichum isolates at the majority of sites. Partial sequences of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of 97 Colletotrichum isolates were amplified as an initial measure of genetic diversity. Phylogenetic analysis with a subset of 22 isolates were performed based on a multi-locus dataset (ACT, TUB2, CAL, CHS-1, GAPDH, ITS) followed by Apn2/MAT IGS sequence-analysis for isolates within the C. gloeosporioides species complex. Molecular analysis associated with phenotypic characteristics revealed six previously described species [C. asianum, C. cliviae, C. dianesei (syn. C. melanocaulon), C. fructicola, C. karstii and C. tropicale] and one new species. This new species is introduced as C. endomangiferae. All species isolated were pathogenic on mango fruits but varied in their virulence. There was no distribution pattern of species among sites and plant tissues, although C. asianum was the most prevalent species at all sites and in all plant tissues studied. Five previously reported Colletotrichum species causing anthracnose in mango fruits in northeastern Brazil were also recovered as endophytes.     

Chronic exposure to low concentrations of strontium 90 affects bone physiology but not the hematopoietic system in mice

The aim of this work was to delineate the effects of chronic ingestion of strontium 90 (⁹⁰Sr) at low concentrations on the hematopoiesis and the bone physiology. A mouse model was used for that purpose. Parent animals ingested water containing 20 kBq l^?1 of ⁹⁰Sr two weeks before mating. Offspring were then continuously contaminated with ⁹⁰Sr through placental transfer during fetal life, through lactation after birth and through drinking water after weaning. At various ages between birth and 20 weeks, animals were tested for hematopoietic parameters such as blood cell counts, colony forming cells in spleen and bone marrow and cytokine concentrations in the plasma. However, we did not find any modification in ⁹⁰Sr ingesting animals as compared with control animals. By contrast, the analysis of bone physiology showed a modification of gene expression towards bone resorption. This was confirmed by an increase in C‐telopeptide of collagen in the plasma of ⁹⁰Sr ingesting animals as compared with control animals. This modification in bone metabolism was not linked to a modification of the phosphocalcic homeostasis, as measured by calcium, phosphorus, vitamin D and parathyroid hormone in the blood. Overall these results suggest that the chronic ingestion of ⁹⁰Sr at low concentration in the long term may induce modifications in bone metabolism but not in hematopoiesis. Copyright © 2012 John Wiley & Sons, Ltd.     

Non-invasive methods for iron overload evaluation in dysmetabolic patients

IntroductionAlthough hyperferritinemia may reflect the inflammatory status of patients with non-alcoholic fatty liver disease (NAFLD), approximately 33% of hyperferritinemia cases reflect real hepatic iron overload.AimTo evaluate a non-invasive method for assessing mild iron overload in patients with NAFLD using 3T magnetic resonance imaging (MRI) relaxometry, serum hepcidin, and the expression of ferritin subunits.MethodsThis cross-sectional study assessed patients with biopsy-proven NAFLD. MRI relaxometry was performed using a 3T scanner in all patients, and the results were compared with iron content determined by liver biopsy. Ferritin, hepcidin, and ferritin subunits were assessed and classified according to ferritin levels and to siderosis identified by liver biopsy.ResultsA total of 67 patients with NAFLD were included in the study. MRI revealed mild iron overload in all patients (sensitivity, 73.5%; specificity, 70%). For mild (grade 1) siderosis, the transverse relaxation rate (R₂*) threshold was 58.9 s^?1 and the mean value was 72.5 s^?1 (SD, 33.9), while for grades 2/3 it was 88.2 s^?1 (SD, 31.9) (p < 0.001). The hepcidin threshold for siderosis was > 30.2 ng/mL (sensitivity, 87%; specificity, 82%). Ferritin H and ferritin L subunits were expressed similarly in patients with NAFLD, regardless of siderosis. There were no significant differences in laboratory test results between the groups, including glucose parameters and liver function tests.ConclusionsMRI relaxometry and serum hepcidin accurately assessed mild iron overload in patients with dysmetabolic iron overload syndrome.     

Known epigenetic biomarkers for prostate cancer detection and management: exploring the potential of blood-based liquid biopsies

Introduction: Although prostate cancer (PCa) stands as an important cause of cancer-related deaths, a sizeable proportion of diagnosed cases are clinically insignificant. Hence, novel and more specific biomarkers to identify clinically significant PCa are needed. Liquid biopsies offer the potential to accurately identify cancer markers, including PCa. Epigenetic biomarkers such as cell-free DNA and circulating RNAs have emerged as minimally invasive cancer markers.

Areas covered: Herein, we provide an overview of epigenetic biomarkers current state based on a comprehensive review of the relevant literature in blood-based liquid biopsies and challenges/limitations of this new and growing field of cancer biomarkers.

Expert opinion: The epigenetic-based biomarkers characteristics make them attractive to the clinics and their minimally invasive assessment are a promising opportunity for PCa detection/management. The main limitations are the lack of robust validation studies and integrated approaches. Future studies would benefit from a change in focus to a ‘selected PCa detection’.     

Expression of cartilage-specific markers in calcified and non-calcified atherosclerotic lesions

Recently, molecular mechanisms resembling endochondral ossification were suggested to be important for atherosclerotic vessel calcification. The aim of this study was to investigate in a series of human atherosclerotic (non-diabetic) lesions of the crural arteries the distribution and expression of classical marker genes of the endochondral ossification pathway. Immunostaining for marker proteins S-100 protein and collagen types II and X were performed on atherosclerotic lesions of different grades (according to Stary). Quantitative real-time PCR for human COL1A1, COL2A1, COL10A1, SOX9, and BMP-2 was applied on RNA isolated from atherosclerotic arteries. In most samples, no expression of collagen type II and S-100 protein was found. Exceptionally, S-100 protein and type II collagen expression was observed very focally within advanced atherosclerotic plaques. Type X collagen was not detected in any of the lesions investigated. Overall, in our study we found no evidence that chondrogenic differentiation pathways are generally active in atherosclerotic plaque formation. In particular type X collagen, one important molecule in cartilage calcification, was not expressed in any of the investigated specimens. Occasionally, however, chondrocytic differentiation markers occur within atherosclerotic lesions. This most likely represents a metaplastic event associated, but not causative for atherosclerotic vessel degeneration and calcification.     

Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis

A Western blot method that uses antigens from culture promastigote forms of Leishmania (Viannia) braziliensis, L. (Leishmania) amazonensis, L. (Leishmania) tropica, and a trypanosomatid (strain 268T) isolated from naturally infected tomatoes was evaluated for laboratory diagnosis of American tegumentary leishmaniasis (ATL). Serum samples were obtained from 108 patients with ATL (group I), 23 chagasic patients (group II), 32 patients with other diseases (group III), and 78 healthy individuals (group IV). The overall analysis showed a sensitivity of 76.90%, 90.40%, 78.50%, and 87.90%, a specificity of 100%, 93.80%, 87.80%, and 77.10%, a positive predictive value of 100%, 94.00%, 89.50%, and 72.50%, a negative predictive value of 75.70%, 90.00%, 75.40%, and 90.20%, and a concordance coefficient kappa of 0.7358, 0.8400, 0.6491, and 0.6287 for L. (V.) braziliensis, L. (L.) amazonensis, L. (L.) tropica, and strain 268T antigens, respectively. The antigenic profile recognized by serum samples from patients with ATL and with Chagas' disease permits serologic distinction between these infections.     

Cytokine participation in the acute rejection of intestinal transplantation in rats

BACKGROUND: Intestinal transplantation is a possible treatment for patients with short bowel syndrome, aiming the reintroduction of oral diet. However, the major obstacle in this procedure is the strong rejection. Delay in rejection diagnosis may be irreversible and lethal. AIM: To define method for early diagnosis of rejection based on the presence of interleucin-6 (IL-6) e interferon- gamma (IFN-gamma) from intestinal allograft. MATERIAL AND METHODS: Isogenic rats Brown-Norway (BN) and Lewis (LEW) were submitted to intestinal heterotopic allotransplantation and divided in two groups: LEW donor to LEW recipient isograft group (C) and BN donor to LEW recipient allograft group (Tx). According to the day of sacrifice, Tx group were subdivided in three subgroups with eight animals each as follow: Tx3--sacrificed at third postoperative day (POD), Tx5--sacrificed at fifth POD and Tx7--sacrificed at seventh POD. Eight animals from control group were subdivided in three moments according to the time of biopsy from the graft as follow: C3--biopsy at third POD; C5--biopsy at fifth POD and C7--biopsy at seventh POD. All animals from control group were sacrificed at seventh POD. Rejection parameters were compared between the control groups (C3 vs C5, C3 vs C7 and C5 vs C7, and allograft group (Tx3 vs Tx5, Tx3 vs Tx7 and Tx5 vs Tx7). The same parameters were analyzed between the control group and allograft groups (C3 vs Tx3, C5 vs Tx5 and C7 vs Tx7). RESULTS: In C group no statistical significant difference regarding the immunoexpression of the cytokines, while in Tx group, immunoexpression of IL-6 and IFN-gamma were remarkable since the fifth postoperative day.