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A new American Onthophagus of the chevrolati species-group is described (Onthophagus bolivari sp. nov.), providing also identification keys to the aztecus complex. Dorsal habitus and genital structures of the new species are illustrated. The relationships and geographical distribution of the aztecus complex are discussed, giving particular attention to the evolutionary history of O. bolivari.

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The Arrau turtle (Podocnemis expansa) is an endangered species, as a result of long-lasting, unsustainable exploitation. To obtain reference haematological values from the wild Podocnemis expansa during post-laying, 20 turtles were captured in the Orinoco River. Blood was obtained from the dorsal cervical sinus in lithium heparin tubes. Red blood cells (RBC), white blood cells (WBC), thrombocytes (TC), packed cell volume (PCV), plasmatic protein (PP), haemoglobin (Hgb), mean corpuscular volume (MCV) and differential leukocyte count were determined. Haematological values were: RBC 0.9×10(9)/L, WBC 5.7×10(9)/L, TC 5.4×10(9)/L, PCV 35.6%, PP 4.2g/dL, Hgb 11.8g/dL, MCV 411fL. The differential leukocyte count comprised: 71% heterophils, 23% lymphocytes, 3% eosinophils, 1.6% basophils, and 1% monocytes. The reports of reference haematology values for the wild P. expansa are limited; therefore, the results presented herein contrast with those values obtained in captivity. This study represents a contribution to the referential haematological values of the wild P. expansa.

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During December 2009 and January 2010, pear trees (Pyrus communis L.) from five orchards located in Allen (Alto Valle) and one in Rio Colorado, both in Rio Negro Province in Argentina, were randomly sampled for Pear blister canker viroid (PBCVd). Ninety-six trees were tested, 20 of cv. Williams, 4 of Abate Fetel, 30 of D'Anjou, and 38 of Packhamn, that showed no symptoms of PBCVd, plus four trees of cv. Red Bartlett that exhibited symptoms of bark pustules and rounded, scaly cankers varying from 2 to 6 cm in diameter on the stems. Purified dsRNA from leaves of 96 trees were analyzed by dot-blot hybridization with a specific probe for PBCVd (2). Of the plants tested, 18 were positive for PBCVd. Three of the positive dsRNAs, with a higher dot-blot signal, were analyzed by electrophoresis in 5% nondenaturing polyacrylamide and a second denaturing polyacrylamide gel. A band, in the portion of viroids migration, was detected with ethidium bromide. The segment corresponding to the three bands was excised and electroeluted. Two-step reverse transcription (RT)-PCR was performed using Moloney-murine leukemia virus (M-MLV) reverse-transcriptase (Promega Corporation, Madison, WI), retrotranscriptase, and PBCVd primers F-5'-GCGGGACAGAAGACGAGGCTCAGGCAGGAAGCAAC-3' and R-5'-TATAAAAGAAAAAAGCGCTTCGGCGGTGCTCGGG-3' (3). The product was legated into pUC 19 vector (Fermentas Inc., Glen Burnie, MD) and cloned following the manufacturer's instructions. Four clones were sequenced by the Unidad de Genómica, Instituto de Biotecnología (Argentina) and the sequences were analyzed with the Lasergene Biocomputing Software for Windows (version 8.0.2; DNASTAR, Madison, WI). The four partial sequences of 296 nucleotides from the local sequences were identical to each other and had the highest nucleotide identity (99.7%) with the Spanish PBCVd (GenBank Accession No. D12823). The local sequence was submitted to GenBank (Accession No. HQ606079). PBCVd is a member of the genus Apscaviroid within the family Pospiviroidae. PBCVd is a 316-nucleotide viroid responsible for pear blister canker disease. It causes pustules, cankers, and/or bark symptoms on the pear indicators A 20 and Fieud 37, whereas infections on most commercial pear cultivars remain symptomless (1). These lesions are entry points for other pathogens to infect the plant. This research indicates the need to test pear propagation material in Argentina, since this is the primary way of spreading this pathogen. To our knowledge, this is the first report of PBCVd in Argentina. References: (1) J. Desvignes et al. Plant Dis. 83:419, 1999. (2) R. Flores et al. J. Gen. Virol. 72:1199, 1991. (3) C. Hernandez et al. J. Gen. Virol. 73:2503, 1992.

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Apple stem pitting virus (ASPV) is an important latent virus of apple trees transmitted by grafting. In pear trees, ASPV is associated with pear vein yellows and pear necrotic spot diseases. Symptoms consist of chlorotic leaf banding and red mottling and flecking along the veins and necrotic spotting in some cultivars may also occur (4). During the spring of 2007, chlorotic leaf banding was observed in Bartlett pear (Pyrus communis L.) trees from one orchard in Bahía Blanca (Buenos Aires Province) and in Anjou, Packham, Abate Fetel, and Bartlett pears in another orchard in General Roca (Río Negro Province). The percentage of symptomatic plants was 10% in both cases. Pooled samples consisting of eight leaves per tree, 25 samples from Bahía Blanca and 25 samples from General Roca, were tested by double-antibody sandwich (DAS)-ELISA with a polyclonal antiserum from BIOREBA AG (Reinach, Switzerland). Five samples from Bahía Blanca and ten from General Roca were positive by DAS-ELISA. Only four positive samples by DAS-ELISA were also positive by immunocapture-reverse transcription (RT)-PCR using virions trapped in a microcentrifuge tube (3). A fragment of 370 bp was amplified with specific primers from each of these four samples. Amplicons were cloned and the nucleotide sequences were determined for one clone of each sample (GenBank Accession Nos. GQ356781, GQ356782, GQ356783, and GQ356784). All sequences had the highest identities with coat protein genes of ASPV. One of them was 94% identical with the coat protein gene of isolate PA66 isolate from Germany (GenBank Accession No. D21829.1) (1). Losses in pear by ASPV have not been demonstrated yet in Argentina. However, when the virus is present with other virus or virus-like diseases, a synergistic effect may occur and growth reduction may exceed 50% (2). Because of the mild symptoms in pear plants, nurserymen or growers must take care when they select material for propagation, in part because laws requiring virus-free propagation material do not exist in Argentina. To our knowledge, this is the first report of ASPV in pears in Argentina. References: (1) W. Jelkmann. J. Gen. Virol. 75:1535, 1994. (2) A. L. Jones and H. S. Aldwinckle. Compendium of Apple and Pear Diseases. The American Phytopathological Society, St. Paul, MN, 1990. (3) W. Menzel et al. J. Virol. Methods 99:81, 2002. (4) M. Németh. Virus, Mycoplasma and Rickettsia Disease of Fruit Trees. Martinus Nijhoff Publishers, Dordrecht, the Netherlands, 1986.

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Sharka disease, caused by Plum pox virus (PPV), is probably the most important disease of stone fruits crops worldwide because of tremendous yield losses from infected trees (1). During November 2004, symptoms resembling sharka disease were observed in a plum and apricot orchard consisting of 5,000 trees in Pocito, San Juan Province, Argentina. Apricot leaves showed chlorotic spots while plum leaves showed chlorotic rings, spots, and irregular edges. Plum fruits were deformed and much smaller than those from symptomless trees. Samples collected from 70 symptomatic trees were tested using double-antibody sandwich enzyme-linked immunosorbent assays with a polyclonal antiserum anti-PPV from BIOREBA (Reinach BL1, Switzerland), and immunosorbent electron microscopy with a polyclonal antiserum from our laboratory made against a recombinant PPV capsid protein (CP). The samples were also tested using double-antibody sandwich indirect enzyme-linked immunosorbent assay using the REAL kit (Durviz, Valencia, Spain) with two different monoclonal antibodies including Mab 5b that recognizes all strains of PPV and Mab 4DG5 that is specific for PPV strain D. Samples were positive with both antibodies in 80% of the cases. Leaf extracts from symptomatic plum samples were also analyzed by immuno-capture reverse-transcription polymerase chain reaction. A 1,209-bp fragment was amplified with specific primers that anneal at the 5' end of the coat protein coding region and the viral 3' end poly A tail. The amplified fragment was cloned and the nucleotide sequence was determined for two of the resulting clones (Gen-Bank Accession Nos. DQ299537 and DQ299538). The sequences were 98% identical with the PPV-strain D from the United States (GenBank Accession No. AF360579) and Germany (GenBank Accession No. X81081). The restriction sites for AluI and RsaI, previously described (2) as typical for the PPV-D strain, were present in the expected positions. To our knowledge, this is the first report of PPV-D in Argentina. Reference: (1) M. Németh. Virus, Mycoplasma, and Rickettsia Disease of Fruit Trees. Martinus Nijhoff Publishers, Dordrecht, the Netherlands, 1986. (2) T. Wetzel et al. J. Virol. Methods 33:355, 1991.

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INTRODUCTION: 2-Chlorodeoxyadenosine (cladribine or 2-CdA) is a purine analogue that has been used successfully in hairy cell leukaemia (HCL). Moreover, it has been increasingly used to treat chronic lymphoproliferative syndromes and paediatric acute myeloid leukaemia. Cutaneous side-effects associated with this drug have seldom been described in cases of HCL. PATIENTS AND METHODS: We describe three patients with chronic lymphocytic leukaemia that presented generalized skin eruptions after treatment with 2-CdA. RESULTS: All patients had advanced disease, receiving 2-CdA as a second or third line chemotherapy. Skin lesions were severe and chemotherapy had to be discontinued. Histological examination of skin biopsies showed an eosinophil-rich infiltrate with flame figures, similar to what is observed in Wells' syndrome (eosinophilic cellulitis). Corticosteroids were effective to control the eruptions. CONCLUSIONS: Cutaneous adverse reactions associated with 2-CdA have seldom been observed in the treatment of HCL. However, as this purine analogue has been used in more advanced cases these may be more frequent and severe. The pathophysiology of these lesions is unclear, but it is probably related to drug-induced change in T-cell imbalance in severely immunosuppressed patients.

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In order to assess the molecular epidemiology of HIV-1 in two neighboring cities located near the epicenter of the HIV-1 epidemics in Brazil (Santos and São Paulo), we investigated 83 HIV-1 strains obtained from samples collected in 1995 from intravenous drug users. The V3 through V5 region of the envelope of gp 120 was analyzed by heteroduplex mobility analysis. Of the 95 samples, 12 (12.6%) were PCR negative (6 samples from each group); low DNA concentration was the reason for non-amplification in half of these cases. Of the 42 typed cases from São Paulo, 34 (81%, 95% confidence limits 74.9 to 87.0%) were B and 8 (19%, 95% confidence limits 12.9 to 25.0%) were F, whereas of the 41 typed cases from Santos, 39 (95%, 95% confidence limits 91.6 to 98.4%) were B and 2 (5%, 95% confidence limits 1.6 to 8.4%) were C. We therefore confirm the relationship between clade F and intravenous drug use in São Paulo, and the presence of clade C in Santos. The fact that different genetic subtypes of HIV-1 are co-circulating indicates a need for continuous surveillance for these subtypes as well as for recombinant viruses in Brazil.

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In order to assess the molecular epidemiology of HIV-1 in two neighboring cities located near the epicenter of the HIV-1 epidemics in Brazil (Santos and São Paulo), we investigated 83 HIV-1 strains obtained from samples collected in 1995 from intravenous drug users. The V3 through V5 region of the envelope of gp 120 was analyzed by heteroduplex mobility analysis. Of the 95 samples, 12 (12.6 percent) were PCR negative (6 samples from each group); low DNA concentration was the reason for non-amplification in half of these cases. Of the 42 typed cases from São Paulo, 34 (81 percent, 95 percent confidence limits 74.9 to 87.0 percent) were B and 8 (19 percent, 95 percent confidence limits 12.9 to 25.0 percent) were F, whereas of the 41 typed cases from Santos, 39 (95 percent, 95 percent confidence limits 91.6 to 98.4 percent) were B and 2 (5 percent, 95 percent confidence limits 1.6 to 8.4 percent) were C. We therefore confirm the relationship between clade F and intravenous drug use in São Paulo, and the presence of clade C in Santos. The fact that different genetic subtypes of HIV-1 are co-circulating indicates a need for continuous surveillance for these subtypes as well as for recombinant viruses in Brazil

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CONTEXT: Young patients affected by acute myeloid leukemia (AML) achieve complete remission (CR) using conventional chemotherapy in about 55-85%. However, 30% of patients fail to achieve CR and the remission duration is often only about 12 months. More intensive treatment after CR seems to be necessary in order to maintain CR and obtain a definitive cure. In Brazil, few reports have been published on this important subject. OBJECTIVE: The aim of this study was to describe a Brazilian experience in the treatment of "de novo" acute myeloid leukemia (AML) in younger adult patients (age < 60 years). DESIGN: Retrospective analysis. SETTING: University Hospital, Hematology and Hemotherapy Center, State University of Campinas, Brazil. PARTICIPANTS: Newly diagnosed cases of "de novo" AML in the period from January 1994 to December 1998 were evaluated retrospectively, in relation to response to treatment, overall survival (OS) and disease free survival (DFS). Cases with acute promyelocytic leukemia (APL) were also included in this analysis. RESULTS: On the basis of an intention to treat, 78 cases of AML, including 17 cases of APL, were evaluated. The overall median follow-up was 272 days. The complete remission (CR) rate was 63.6% in the AML group (excluding APL) and 78% in the APL group. The 5-year estimated disease-free survival (DFS) was 80% for the APL group and 34% for the AML group (P = 0.02). The 5-year estimated overall survival (OS) was 52% for the APL group and 20.5% for the AML group, respectively (P = NS). Relapse was observed in 12/39 (30.7%) patients with AML and 1/11 (9%) with APL. CONCLUSIONS: These results are similar to those reported in the literature. However, relapse and mortality rates remain high, and a search for more aggressive strategies in order to prevent relapse is recommended.

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CONTEXT: Liver damage is relatively common in patients affected by Hodgkin's disease. A smaller proportion of cases develops jaundice. Recently, the vanishing bile duct syndrome was described in Hodgkin's disease. The mechanisms of this severe complication have been poorly understood until now. OBJECTIVE: To describe a rare case of intra-hepatic cholestasis due to vanishing bile duct syndrome. DESIGN: Case report. CASE REPORT: A 38-year-old male patient affected by Hodgkin's disease. Liver biopsy showed no detectable Hodgkin's disease. Intra-hepatic cholestasis was found and none of the six portal tracts analyzed contained normal bile ducts. The treatment was based on conventional and high-dose escalation chemotherapy. The patient died from an irreversible liver failure while in complete remission from Hodgkin's disease.

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