Anethole Prevents the Alterations Produced by Diabetes Mellitus in the Sciatic Nerve of Rats.

Anethole is a terpenoid with antioxidant, anti-inflammatory, and neuronal blockade effects, and the present work was undertaken to study the neuroprotective activity of anethole against diabetes mellitus (DM)-induced neuropathy. Streptozotocin-induced DM rats were used to investigate the effects of anethole treatment on morphological, electrophysiological, and biochemical alterations of the sciatic nerve (SN). Anethole partially prevented the mechanical hyposensitivity caused by DM and fully prevented the DM-induced decrease in the cross-sectional area of the SN. In relation to electrophysiological properties of SN fibers, DM reduced the frequency of occurrence of the 3rd component of the compound action potential (CAP) by 15%. It also significantly reduced the conduction velocity of the 1st and 2nd CAP components from 104.6 ± 3.47 and 39.8 ± 1.02 to 89.9 ± 3.03 and 35.4 ± 1.56 m/s, respectively, and increased the duration of the 2nd CAP component from 0.66 ± 0.04 to 0.82 ± 0.09 ms. DM also increases oxidative stress in the SN, altering values related to thiol, TBARS, SOD, and CAT activities. Anethole was capable of fully preventing all these DM electrophysiological and biochemical alterations in the nerve. Thus, the magnitude of the DM-induced neural effects seen in this work, and the prevention afforded by anethole treatment, place this compound in a very favorable position as a potential therapeutic agent for treating diabetic peripheral neuropathy.

The Essential Oil of Hyptis crenata Inhibits the Increase in Secretion of Inflammatory Mediators.

Background: Hyptis crenata is a plant of great ethnopharmacological importance widely distributed in South American countries. In Northeast Brazil, teas or infusions of its aerial parts are used in folk medicine to treat several acute and chronic inflammatory diseases. In a previous work we have demonstrated that the essential oil of H. crenata (EOHc) has an antiedematogenic effect. The aim of this work was to evaluate the effect of EOHc on cytokines secretion and cellular infiltration. Methods: Peritonitis and paw edema models induced by carrageenan were used to determine leucocyte count, myeloperoxidase (MPO) activity, nitrite, and cytokines secretion. Results: EOHc (10−300 mg/kg) significantly inhibited leucocyte migration and reduced the neutrophil count (control: 1.46 × 103 ± 0.031 × 103/mL) of the total leucocytes population in extracellular exudate (control: 2.14 × 103 ± 0.149 × 103/mL) by 15.00%, 43.29%, 65.52%, and 72.83% for the doses of 10, 30, 100, and 300 mg/kg EOHc, respectively (EC50: 24.15 mg/kg). EOHc (100 mg/kg) inhibited the increase in myeloperoxidase activity and completely blocked the increase in nitrite concentration induced by carrageenan. EOHc markedly reduced the pro-inflammatory cytokines (IL-6, MCP-1, IFN-γ, TNF-α, and IL-12p70) and increased IL-10, an anti-inflammatory cytokine (compared to control group, p < 0.05). Conclusions: This study demonstrates that EOHc has a long-lasting anti-inflammatory effect mediated through interference on MPO activity, and nitrite, and cytokines secretion. This effect, coupled with low EOHc toxicity, as far as results obtained in mice could be translated to humans, suggests that EOHc has great potentiality as a therapeutic agent.

Ground-foraging ants (Hymenoptera: Formicidae) and rainfall effect on pitfall trapping in a deciduous thorn woodland (caatinga), Northeastern Brazil.

The semi-arid Caatinga is the fourth largest biome of Brazil, which biota still remains one of the most poorly known, especially with regard to invertebrate groups. In this study, a ground-foraging ant assemblage was surveyed during one year and the effect of rainfall on pitfall trapping was assessed. The study was performed in an area located in the municipality of Pentecoste (3 degrees 48' S - 39 degrees 20' W), in the State of Ceará. A 200m transect with 20 equidistant sampling points was established. Transect sampling was performed once a month during 12 months, over the period August 2008-August 2009. At each sampling point, a pitfall trap partially filled with a mixture of ethanol and monoethylene glycol was placed at the beginning of each month and remained in the field for seven days. 39 species belonging to six subfamilies and 19 genera, plus two unidentified species, were collected, with Pheidole (10 spp.) and Camponotus (8 spp.) being the taxa with the most species. 23 species were frequent, being found in more than 50% of the 12 transect samplings. Five species had an intermediate frequency (25 to 50%), while 13 were relatively infrequent (less than 25%). Most of the species (22) showed low occurrence, being found in less than 10% of the 240 samples (20 samples each month, during 12 months). Only five species were collected in more than 50% of the samples, those species being also responsible for most of the total abundance (number of captured individuals of all species) observed each month. The species-accumulation curves (observed and estimated) indicated that sampling sufficiency was attained, and that about 92% of the estimated ground-foraging ant fauna had been collected. 40 and 29 species were collected in the dry and rainy season, respectively, with monthly species richness ranging from 13 to 28. The total ant abundance showed a drastic decrease during the rainy season, and a negative linear correlation was found between rainfall and total ant abundance (R2 = 0.68). A similar negative linear correlation was found for species occurrences against rainfall (R2 = 0.71), and for mean number of species per pitfall trap against rainfall (R2 = 0.71). However, some species showed equal abundance, occurrence and mean number of individuals per pitfall trap in both seasons, while others showed a much higher abundance and occurrence during the rainy season. Pitfall trapping as a method to sample ground-foraging ant assemblage of the Caatinga biome and potential factors responsible for lower pitfall trap performance during rainy season are discussed.

Effect of an aqueous extract of Chrysobalanus icaco on the adiposity of Wistar rats fed a high-fat diet.

INTRODUCTION: Objective: the purpose of this study was to investigate the effects of Chrysobalanus icaco on adiposity and its mechanism of action in the gene and protein expression of acetyl-CoA carboxylase (ACC), a key enzyme in lipogenesis. Method: Wistar rats were divided into a regular or control group (CG) and a high-fat diet (HFD) group. HFD was treated with saline or aqueous extract of Chrysobalanus icaco (AECI) for four weeks. Body weight and food intake were assessed. Subcutaneous, retroperitoneal and periepididymal adipose tissue samples were collected and weighed. Adipocytes from periepididymal tissue were isolated and analyzed. The gene and protein expression of ACC in subcutaneous tissue was determined. Results: AECI showed no effect on intake or body weight. However, the weight of the fat pads and the gene and protein expression of ACC were lower, and glucose tolerance was improved. Conclusion: the aqueous extract of Chrysobalanus icaco proved beneficial for the treatment of obesity, preventing fat storage and improving glycemic homeostasis.

INTRODUCCIÓN: Objetivo: el objetivo de este estudio fue investigar los efectos del extracto acuoso de Chrysobalanus icaco (AECI) en la adiposidad y su mecanismo de acción en la expresión génica y proteica de la acetil-CoA-carboxilasa (ACC), una enzima clave para la lipogénesis. Métodos: se usaron ratones macho Wistar que se asignaron a una dieta estándar de control (CG) o a una rica en grasa (HFD). La HFD se trató con solución salina o con extracto acuoso de Chrysobalanus icaco (AECI) durante cuatro semanas. Se evaluaron el peso corporal y el consumo alimentario. Se aislaron y analizaron muestras de tejido adiposo subcutáneo, retroperitoneal y periepididímico. Se determinó la expresión génica y proteica de ACC en el tejido subcutáneo. Resultados: el AECI no mostró ningún efecto sobre la ingesta de alimento y tampoco sobre el peso corporal. Sin embargo, el tratamiento con AECI redujo el peso de los tejidos adiposos y la expresión génica y proteica de ACC, y mejoró también la tolerancia a la glucosa. Conclusión: Chrysobalanus icaco (AECI) resultó ser beneficioso para el tratamiento de la obesidad, previniendo el almacenamiento de grasa y mejorando la homeostasis glucémica.

Hepatoprotective Effect of Essential Oils from Hyptis crenata in Sepsis-Induced Liver Dysfunction.

No specific therapeutics are available for the treatment of sepsis-induced liver dysfunction, a clinical complication strongly associated with the high mortality rate of septic patients. This study investigated the effect of the essential oil of Hyptis crenata (EOHc), a lamiaceae plant used to treat liver disturbances in Brazilian folk medicine, on liver function during early sepsis. Sepsis was induced by the cecal ligation and puncture (CLP) model. Rats were divided into four groups: Sham, Sham+EOHc, CLP, and CLP+EOHc. EOHc (300 mg/kg) was orally administered 12 and 24 h after surgery. The animals were sacrificed for blood collection and liver tissue samples 48 h after surgery. Hepatic function was evaluated by measuring serum bilirubin, alkaline phosphatase (ALP), aspartate aminotransferase, and alanine aminotransferase (ALT) levels. The levels of malondialdehyde and the activity of superoxide dismutase, catalase, and GSH peroxidase (GSH-Px) were measured for assessment of oxidative stress. Liver morphology was analyzed by hematoxylin and eosin staining. EOHc normalized serum ALP, ALT, and bilirubin levels and inhibited morphological changes. In addition, we observed that EOHc inhibited elevation in hepatic lipid peroxidation and reduction of the glutathione peroxidase activity induced by sepsis. Our data show that EOHc plays a protective effect against liver injury induced by sepsis.

Diagnosis and treatment of chronic lymphocytic leukemia: recommendations from the Brazilian Group of Chronic Lymphocytic Leukemia.

Chronic lymphocytic leukemia is characterized by clonal proliferation and progressive accumulation of B-cell lymphocytes that typically express CD19+, CD5+ and CD23+. The lymphocytes usually infiltrate the bone marrow, peripheral blood, lymph nodes, and spleen. The diagnosis is established by immunophenotyping circulating B-lymphocytes, and prognosis is defined by two staging systems (Rai and Binet) established by physical examination and blood counts, as well as by several biological and genetic markers. In this update, we present the recommendations from the Brazilian Group of Chronic Lymphocytic Leukemia for the diagnosis and treatment of chronic lymphocytic leukemia. The following recommendations are based on an extensive literature review with the aim of contributing to more uniform patient care in Brazil and possibly in other countries with a similar social-economic profile.

Binet's staging system and VH genes are independent but complementary prognostic indicators in chronic lymphocytic leukemia.

PURPOSE: Rai's and Binet's staging systems have contributed significantly to the identification of major prognostic groups in chronic lymphocytic leukemia (CLL), though they fail to accurately predict disease progression at the individual level. Biologic factors, such as the mutational status of the immunoglobulin heavy-chain variable genes (VH, cytogenetics, CD38 expression, and some serum markers, have recently improved prognostic assessment in CLL. In this study, we analyzed the prognostic value of VH mutational status within the different stages of Binet's classification in 145 patients. PATIENTS AND METHODS: Our series consisted of 83 VH mutated (MT) and 62 VH unmutated (UM) patients. MT cases predominated within Binet's stage A (70%), whereas UM cases predominated among stages B and C (62%). RESULTS: Median overall survival (OS) was 84 months for UM patients and was not achieved for the MT group (70% 12-year survival, P <.0001). Concerning Binet's stage A, both median OS and progression-free survival were significantly shorter for UM patients when compared with those of MT patients (97 months v not achieved, P =.0017; and 42 v 156 months, P <.0001), which compared favorably with the classical A' and A" substaging. The VH mutational profile could also segregate stage B and C patients into two groups with different survival patterns (median OS, 78 v 120 months for UM and MT patients, respectively; P =.002). CONCLUSION: The significant survival differences observed between the VH mutational groups, among stage A and stage B and C patients, indicate that Binet's classification and VH genes are independent prognostic variables and are most likely complementary.

Effect of an aqueous extract of Chrysobalanus icaco on the adiposity of Wistar rats fed a high-fat diet / Efecto del extracto acuoso de Chrysobalanus icaco sobre la adiposidad de ratas Wistar alimentadas con una dieta alta en grasas

OBJECTIVE: the purpose of this study was to investigate the effects of Chrysobalanus icaco on adiposity and its mechanism of action in the gene and protein expression of acetyl-CoA carboxylase (ACC), a key enzyme in lipogenesis. METHOD: Wistar rats were divided into a regular or control group (CG) and a high-fat diet (HFD) group. HFD was treated with saline or aqueous extract of Chrysobalanus icaco (AECI) for four weeks. Body weight and food intake were assessed. Subcutaneous, retroperitoneal and periepididymal adipose tissue samples were collected and weighed. Adipocytes from periepididymal tissue were isolated and analyzed. The gene and protein expression of ACC in subcutaneous tissue was determined. RESULTS: AECI showed no effect on intake or body weight. However, the weight of the fat pads and the gene and protein expression of ACC were lower, and glucose tolerance was improved. CONCLUSION: the aqueous extract of Chrysobalanus icaco proved beneficial for the treatment of obesity, preventing fat storage and improving glycemic homeostasis

OBJETIVO: el objetivo de este estudio fue investigar los efectos del extracto acuoso de Chrysobalanus icaco (AECI) en la adiposidad y su mecanismo de acción en la expresión génica y proteica de la acetil-CoA-carboxilasa (ACC), una enzima clave para la lipogénesis. MÉTODOS: se usaron ratones macho Wistar que se asignaron a una dieta estándar de control (CG) o a una rica en grasa (HFD). La HFD se trató con solución salina o con extracto acuoso de Chrysobalanus icaco (AECI) durante cuatro semanas. Se evaluaron el peso corporal y el consumo alimentario. Se aislaron y analizaron muestras de tejido adiposo subcutáneo, retroperitoneal y periepididímico. Se determinó la expresión génica y proteica de ACC en el tejido subcutáneo. RESULTADOS: el AECI no mostró ningún efecto sobre la ingesta de alimento y tampoco sobre el peso corporal. Sin embargo, el tratamiento con AECI redujo el peso de los tejidos adiposos y la expresión génica y proteica de ACC, y mejoró también la tolerancia a la glucosa. CONCLUSIÓN: Chrysobalanus icaco (AECI) resultó ser beneficioso para el tratamiento de la obesidad, previniendo el almacenamiento de grasa y mejorando la homeostasis glucémica

Volatile oil of Croton zehntneri per oral sub-acute treatment offers small toxicity: perspective of therapeutic use

ABSTRACT Croton zehntneri Pax & K. Hoffm., Euphorbiaceae, or "canela-de-cunhé" is used in the Northeast Brazil to treat several diseases. Leaves and aerial parts of C. zehntneri are rich in volatile oil of high potential therapeutic. This study aimed to investigate volatile oil systemic toxicity after per oral treatment in rats. Volatile oil characterization (gas chromatography and mass spectrometry) showed 85.7% anethole and 4.8% estragole. Male Wistar rats (116-149 g) were treated with volatile oil (250 mg/kg p.o.) during ten weeks and evaluated for the following parameters: survival; food and water intake; body mass; absolute/relative organs weight; hemogram; plasma biochemical dosage; organs morphology. Volatile oil did not alter animal water and food consumption or the relative/absolute weight of most organs, but animals gained less weight. Volatile oil did not alter function biomarkers of pancreas, kidney, heart or liver, but increased plasma gamma-glutamyltranspeptidase (liver biomarker) and decreased uric acid (kidney biomarker). Although volatile oil had caused discrete morphological alterations in some organs, it did not induce architectural changes in these organs. In conclusion, the sub-acute per oral treatment with volatile oil no longer than ten weeks in rats offers small toxicity at doses below 250 mg/kg.

Gastroprotective effects of the essential oil of Hyptis crenata Pohl ex Benth. on gastric ulcer models.

ETHNOPHARMACOLOGICAL RELEVANCE: Hyptis crenata Pohl ex Benth (Lamiaceae), popularly known as "hortelã-brava" or "hortelã do campo", is widely distributed in the northeast of Brazil. In Brazil, the leaves of this plant have been used in traditional medicine for the treatment of gastrointestinal disturbances, including gastric ulcers. In an attempt to experimentally validate this claimed antiulcerogenic activity, the gastroprotective effects of the essential oil extracted from the leaves of the Hyptis crenata Pohl ex Benth (EOHc) were evaluated in recognized gastric ulcer models in mice. MATERIALS AND METHODS: EOHc was obtained by hydrodistillation and analyzed using gas chromatography-mass spectrometry (GC-MS). Swiss male mice (25-30g) were used for the studies. The gastric ulcers were induced by oral administration of absolute ethanol or indomethacin 45min after oral pretreatment with EOHc, vehicle and positive control drugs. One hour after the ulcerative challenges, the stomachs were removed and the area of the lesions was measured. The volume, pH and total acidity of the gastric secretions were determined using the pylorus ligature model. The gastrointestinal motility was measured using gastric emptying and intestinal transit. The ethanol-induced gastric mucus depletion and lipid peroxidation were also analyzed. RESULTS: Our findings are as follows: A significant inhibition of gastric lesions induced by absolute ethanol was observed in the mice pre-treated with EOHc, at a dose of 30 and 100 and 300mg/kg (5.56±1.51, 2.88±0.82 and 1.71±0.54mm(2), respectively) compared to control group (118.03±35.4mm(2)). Also, EOHc (300mg/kg) produced a gastroprotective effect against the gastric lesions induced by indomethacin (16.07±4.68mm(2)) compared to control group (38.64±6.1mm(2)). EOHc pretreatment produced a reduction in the ethanol-induced lipid peroxidation from 3.9±0.22 to 2.4±0.1µmol/mg tissue (EOHc-300mg/kg and control group, respectively). We also observed that EOHc pretreatment decreased the gastric emptying, but did not alter the intestinal transit ratio, ethanol-induced depletion of the gastric wall mucus or secretion parameters (volume, pH and [H(+)]). CONCLUSION: Our data indicate that EOHc exerts a gastroprotective effect, indicated by its significant inhibition of gastric lesions in ethanol- and indomethacin-induced ulcer models, which may be associated with its accelerating effect on gastric emptying and reduction in oxidative damages. Our data suggest a potential therapeutic application for EOHc in the treatment of gastric ulcers.

Diagnosis and treatment of chronic lymphocytic leukemia: recommendations from the Brazilian Group of Chronic Lymphocytic Leukemia

ABSTRACT Chronic lymphocytic leukemia is characterized by clonal proliferation and progressive accumulation of B-cell lymphocytes that typically express CD19+, CD5+ and CD23+. The lymphocytes usually infiltrate the bone marrow, peripheral blood, lymph nodes, and spleen. The diagnosis is established by immunophenotyping circulating B-lymphocytes, and prognosis is defined by two staging systems (Rai and Binet) established by physical examination and blood counts, as well as by several biological and genetic markers. In this update, we present the recommendations from the Brazilian Group of Chronic Lymphocytic Leukemia for the diagnosis and treatment of chronic lymphocytic leukemia. The following recommendations are based on an extensive literature review with the aim of contributing to more uniform patient care in Brazil and possibly in other countries with a similar social-economic profile.

Ground-foraging ants (Hymenoptera: Formicidae) and rainfall effect on pitfall trapping in a deciduous thorn woodland (Caatinga), Northeastern Brazil

The semi-arid Caatinga is the fourth largest biome of Brazil, which biota still remains one of the most poorly known, especially with regard to invertebrate groups. In this study, a ground-foraging ant assemblage was surveyed during one year and the effect of rainfall on pitfall trapping was assessed. The study was performed in an area located in the municipality of Pentecoste (3º48’ S - 39º20’ W), in the State of Ceará. A 200m transect with 20 equidistant sampling points was established. Transect sampling was performed once a month during 12 months, over the period August 2008-August 2009. At each sampling point, a pitfall trap partially filled with a mixture of ethanol and monoethylene glycol was placed at the beginning of each month and remained in the field for seven days. 39 species belonging to six subfamilies and 19 genera, plus two unidentified species, were collected, with Pheidole (10 spp.) and Camponotus (8 spp.) being the taxa with the most species. 23 species were frequent, being found in more than 50% of the 12 transect samplings. Five species had an intermediate frequency (25 to 50%), while 13 were relatively infrequent (less than 25%). Most of the species (22) showed low occurrence, being found in less than 10% of the 240 samples (20 samples each month, during 12 months). Only five species were collected in more than 50% of the samples, those species being also responsible for most of the total abundance (number of captured individuals of all species) observed each month. The speciesaccumulation curves (observed and estimated) indicated that sampling sufficiency was attained, and that about 92% of the estimated ground-foraging ant fauna had been collected. 40 and 29 species were collected in the dry and rainy season, respectively, with monthly species richness ranging from 13 to 28. The total ant abundance showed a drastic decrease during the rainy season, and a negative linear correlation was found between rainfall and total ant abundance (R2=0.68). A similar negative linear correlation was found for species occurrences against rainfall (R2=0.71), and for mean number of species per pitfall trap against rainfall (R2=0.71). However, some species showed equal abundance, occurrence and mean number of individuals per pitfall trap in both seasons, while others showed a much higher abundance and occurrence during the rainy season. Pitfall trapping as a method to sample ground-foraging ant assemblage of the Caatinga biome and potential factors responsible for lower pitfall trap performance during rainy season are discussed. Rev. Biol. Trop. 59 (4): 1637-1650. Epub 2011 December 01.

La Caatinga semiárida es el cuarto bioma más grande de Brasil. Pese a ello, es también el bioma brasileño cuya biota continúa siendo la más pobremente estudiada, especialmente en lo que se refiere a grupos de invertebrados. En este estudio se examinó durante un año el grupo de hormigas que forrajean en el suelo de un área de Caatinga y se evaluó el efecto de la lluvia sobre las trampas de caída. El estudio se llevó a cabo en un área del municipio de Pentecoste (3º48’ S - 39º20’ W), estado de Ceará. Se estableció un transecto de 200m con 20 puntos de muestreo equidistantes. El muestreo del transecto se realizó mensualmente durante 12 meses, entre Agosto 2008-Agosto 2009. En cada punto de muestreo se colocó al principio de cada mes una trampa de caída parcialmente llena con una mezcla de etanol y monoetilenglicol y se mantuvo en el campo durante siete días. Se recogieron 39 especies pertenecientes a seis subfamilias y 19 géneros, además de dos especies sin identificar, siendo Pheidole (10 spp) y Camponotus (8 spp) los taxones con más especies. Veintitrés especies fueron frecuentes, se registraron en más del 50% de los 12 transectos muestreados. Cinco especies tuvieron una frecuencia intermedia (25 a 50%), mientras 13 fueron relativamente infrecuentes (menos del 25%). La mayoría de las especies (22) mostraron una presencia baja, encontrándose en menos del 10% de las 240 muestras (20 muestras cada mes durante 12 meses). Sólo cinco especies fueron recogidas en más del 50% de las muestras, fueron además responsables de casi toda la abundancia total (número de individuos capturados de todas las especies) mensual. Las curvas de acumulación de especies (observadas y estimadas) indicaron que se consiguió un muestreo suficiente y que se había recogido cerca del 92% de la fauna estimada de hormigas terrícolas forrajeras. Se recogieron 40 y 29 especies durante las estaciones seca y lluviosa, respectivamente, con una riqueza de especies mensual entre 13 y 28. La abundancia total de especies mostró una disminución drástica durante la estación de lluvias, y se encontró una correlación linear negativa entre la pluviosidad y la abundancia total de hormigas (R2=0.68). Una correlación linear negativa similar se encontró entre la ocurrencia de especies y la pluviosidad (R2=0.71), y entre el número medio de especies por trampa de caída y la pluviosidad (R2=0.71). Sin embargo, mientras se observó que algunas especies tenían la misma abundancia, presencia, y número medio de individuos por trampa de caída en ambas estaciones, otras tenían una abundancia y presencia mucho mayor durante la estación lluviosa. Se discute el uso de trampas de caída como método para muestrear el grupo de hormigas que forrajean en el suelo del bioma de la Caatinga, así como los factores potenciales responsables del rendimiento más bajo de las trampas de caída durante la estación lluviosa.

The LPL/ADAM29 expression ratio is a novel prognosis indicator in chronic lymphocytic leukemia.

Although the zeta-associated protein of 70 kDa (ZAP-70) is overexpressed in patients with chronic lymphocytic leukemia (CLL) displaying unmutated IGVH genes and poor prognosis, a previous microarray study from our group identified overexpression of LPL and ADAM29 genes among unmutated and mutated CLL, respectively. To assess the prognostic value of these genes, we quantified their expression by real-time quantitative polymerase chain reaction (PCR) in a cohort of 127 patients with CLL and correlated this with clinical outcome, IGVH mutational status, and ZAP-70 protein expression. IGVH mutational status, ZAP-70, and the LPL and ADAM29 mRNA ratios (L/A ratio) were predictive of event-free survival for the whole cohort and for patients with stage A disease. In patients in stage B and C, the L/A ratio was an independent prognostic factor, whereas ZAP-70 did not predict survival. Simultaneous usage of the L/A ratio and ZAP-70 expression allowed an almost perfect (99%) assessment of the IGVH status in the 80% of patients with concordant results (L/A+, ZAP-70+ or L/A-, ZAP-70-). LPL and ADAM29 gene expression could also be determined by a simple competitive multiplex reverse transcription PCR assay. Overall, quantification of LPL and ADAM29 gene expression is a strong prognostic indicator in CLL, providing better prognostic assessment than ZAP-70 in advanced stages of the disease.

Chronic lymphocytic leukemia B cells expressing AID display dissociation between class switch recombination and somatic hypermutation.

In B cells, somatic hypermutation (SHM) and class switch recombination (CSR) depend on the activation-induced cytidine deaminase (AID) gene product, although the precise mode of action of AID remains unknown. Because some chronic lymphocytic leukemia (CLL) B cells can undergo CSR without SHM, it constitutes a useful model to dissect AID function. In this work, we have studied AID expression, the presence of mutations in the preswitch mu DNA region, CSR, and the SHM in 65 CLL patients. Our results demonstrate that unmutated CLL B cells can constitutively express AID and that AID expression is associated with the presence of mutations in the preswitch region and in clonally related isotype-switched transcripts. They also demonstrate that in CLL without constitutive AID expression, AID induction on stimulation results in preswitch mutations and the CSR process. Our results show a dissociation between SHM and CSR in CLL and suggest that, in this disease, AID would require additional help for carrying out the SHM process.

CLL: chromosomal abnormalities (FISH) and their relation with clinical stage, CD38 and ZAP-70 / Leucemia linfocítica crônica: anormalidades cromossômicas e a sua relação com o estágio clínico CD38 e o ZAP-70

Chronic lymphocytic leukemia is the most prevalent type of leukemia in the West. It is characterized by an extremely variable clinical course. The aim of the study was to detect the most frequent chromosomal abnormalities in patients with CLL using FISH, and assess them regarding age, gender, clinical stage and CD38 and ZAP-70 expressions. We found 51.7 percent of the patients with chromosome abnormalities. The most frequent one was del 13q14 in 34.5 percent of cases. It was associated to other alterations in 17.2 percent. 17p13 deletions were found in 17.2 percent and trisomy 12 in 13.8 percent (in isolation in 6.9 percent and associated to del 13q14, in 6.9 percent of the cases). An 11q22 deletion was found in one case associated to a 13q14 deletion. To better evaluate the relationship between chromosome aberrations and other prognostic factors in CLL, two cytogenetics groups were considered: favorable (13q deletion in isolation and no alteration) and unfavorable outcomes (trisomy 12, 17p13 deletion, 11q22 deletion and two simultaneous alterations).The unfavorable alterations were more frequently seen among young individuals (<60y). There were more females (70 percent) than males in this group (p=0.04). In relation to the Binet's staging system, patients with unfavorable cytogenetic alterations, tended to be B and C stages, while in the favorable group prevailed patients in stage A. Additionally, patients with poor prognostic cytogenetics tended to express CD38 and ZAP-70 proteins.

A leucemia linfocítica crônica (LLC) é o tipo de leucemia mais prevalente no Ocidente e é caracterizada por curso clínico extremamente variável. O objetivo deste estudo foi detectar as anomalias cromossômicas mais freqüentes em pacientes com LLC, empregando a técnica FISH, e correlacioná-las com idade, sexo, estádio clínico, expressão de CD 38 e ZAP-70. Foram encontradas alterações cromossômicas em 51,7 por cento dos pacientes. A mais freqüente foi a del 13q14, observada em 34,5 por cento dos casos e que esteve associada a outras anomalias em 17,2 por cento. Deleção 17p13 foi encontrada em 17,2 por cento e trissomia 12 em 13,8 por cento (isolada em 6,9 por cento e associada à del 13q14 em 6,9 por cento). Deleção 11q22 foi observada em um caso em concomitância à del 13q14. Para melhor avaliar a relação entre alteração cromossômica e outros fatores prognósticos em LLC, dois grupos citogenéticos foram considerados: favorável (deleção 13q isolada e ausência de alterações) e desfavorável (trissomia 12, deleção 17p13, deleção 11q22 e duas anomalias simultâneas). As alterações desfavoráveis foram mais freqüentemente observadas em indivíduos jovens (<60 anos) e em mulheres (70 por cento)(p=0,04). Em relação ao sistema de estadiamento de Binet, houve tendência dos pacientes com alterações cromossômicas desfavoráveis apresenteram-se nos estágios B e C enquanto no grupo favorável prevaleceram aqueles com estágio A. Em adição, pacientes com achados citogenéticos de prognóstico desfavorável tiveram tendência a expressar proteínas CD 38 e ZAP-70.

Marcadores de prognóstico na leucemia linfocítica crônica / Prognostic factors in chronic lymphocytic leukemia

A leucemia linfocítica crônica (LLC) é reconhecida pela evolução clínica heterogênea que não consegue ser prevista com exatidão pelos sistemas de estadiamento clínico a nível individual. Isto levou à investigação de outros marcadores de prognóstico que poderiam agregar valor preditivo aos sistemas de estadiamento ou até mesmo substituí-los. Entre os marcadores clínicos e biológicos inicialmente encontrados, as aberrações cromossômicas e o estado mutacional dos genes de imuðnoglobulinas demonstraram uma alta precisão na avaliação de prognóstico na LLC. No entanto, as técnicas empregadas nestes estudos são laboriosas e inacessíveis à maioria dos serviços de onco-hematologia, o que motivou a busca por marcadores substitutos (surrogate). Entre os potenciais marcadores surrogate, CD38 e Zap-70 possuem um papel independente de prognóstico na LLC, com um poder de predição evolutiva tão (ou mais) preciso quanto o perfil mutacional das imunoglobulinas, possibilitando sua substituição definitiva num futuro próximo. Novos marcadores como LPL, LPL/ADAM29 e Vimentina têm apresentado resultados preliminares bastante atrativos, porém ainda aguardam validação em outras séries de pacientes. Mesmo com a identificação de marcadores biológicos altamente precisos, os sistemas de estadiamento clínico ainda não devem ser abandonados.

Chronic lymphocytic leukemia is characterized by a variable clinical course that cannot be predicted accurately by clinical staging systems in individual patients. This prompted the investigation of other prognostic factors capable of adding predictive power to clinical staging systems or even substituting them. Among the clinical and biological markers found initially, genomic aberrations and the mutational status of immunoglobulin genes demonstrated a high level of prognostic prediction in CLL. However, the techniques employed in these studies are laborious and inaccessible for most hematology-oncology facilities, which motivated the hunt for surrogate markers. Among the potential surrogates, CD38 and Zap-70 play an independent prognostic role in CLL, with a predictive power as precise as (or even better than) the immunoglobulin mutational profile, pointing for its replacement in the near future. Novel factors such as LPL, LPL/ADAM29 and Vimentin have showed quite attractive preliminary results, but still wait for validation in further series of patients. Besides the recognition of powerful biological markers, the clinical staging systems should not be abandoned so far.