Physical activity measured with an accelerometer in dogs following extracapsular stabilisation to treat cranial cruciate ligament rupture.

OBJECTIVES: To quantify the physical activity levels in dogs with cranial cruciate ligament rupture before and after lateral fabellar suture stabilisation surgery. MATERIALS AND METHODS: Seventeen dogs (mean weight, 12.3±5.1 kg) with unilateral cranial cruciate ligament rupture were fitted with an accelerometer for seven consecutive days at four different time points: before surgery (T0), one (T1), three (T3) and six (T6) months after surgery. The total activity and times spent in sedentary activity, light to moderate activity and vigorous activity were recorded by the accelerometer, and preoperative and postoperative data were compared. At all time points, dogs underwent clinical evaluations (lameness score, stifle pain score and thigh circumference) and their owners were asked to respond to questionnaires to subjectively score the physical activity and quality of life of the dogs. RESULTS: At the four time points, the dogs spent between 21.2 and 21.4 hours on sedentary behaviour, 2.3 and 2.5 hours performing light to moderate activity, and 13 to 15 minutes performing vigorous activity. There was no increase in physical activity variables or decrease in sedentary behaviour over time. Lameness scores, pain score and dogs' quality of life improved significantly during the postoperative period. At T6, 17 (100%) of 17 dogs presented no lameness, 16 (94%) of 17 dogs presented no stifle pain, 16 (94%) of 17 owners rated the quality of life as very good and excellent, and 16 (100%) of 16 owners reported a total return to normal activity levels. CLINICAL SIGNIFICANCE: The clinical recovery after extracapsular stabilisation of the stifle joint was not associated with a spontaneous increase in physical activity or a decrease in sedentary behaviour.

Spatial distribution of Ceratitis capitata in guava orchards and influences from orchard management.

The objectives of this research were: (i) to Estimate the quantitative occurrence of Ceratitis capitata captured in McPhail traps in cultivating guava; (ii) to investigate the spatial distribution patterns of C. capitata in guava orchards; (iii) to compare the index of the FTD (fruit fly/trap/day) to the type of spatial distribution of C. capitata with the Negative Binomial to set the best time for control of the population in the context of Integrated Pest Management; (iv) Verify the influence of the pruning, spraying, and mowing on the distributions of medfly in guava. Was used 30 McPhail traps installed in three commercial orchards of guava. The spatial distribution was evaluated by the regression model with the Taylor power method, from the log of variance to the log of the mean number of individuals. Ceratitis capitata has aggregated distribution behavior. The potential risk of economic damage is associated with aggregate distribution behavior. The beginning of this distribution indicates the most appropriate time for using control techniques.

Cisto aracnoide medular em um cão: relato de caso / Arachnoid cyst in a dog: case report

O cisto aracnoide medular (CAM) é uma doença que pode ocorrer em humanos e animais, podendo causar sinais clínicos neurológicos. A origem dessa enfermidade ainda é desconhecida assim como sua patofisiologia. Acredita-se que pode ser congênita ou adquirida. Até o momento, não foi verificada predileção por raça, sexo ou idade. O objetivo deste trabalho é relatar um caso de CAM lombar em um cão com 13 anos de idade, que causou paralisia dos membros pélvicos. Ao exame clínico, o paciente apresentava dor lombar na palpação epaxial, incontinência urinária e fecal, com paraplegia de membros pélvicos. A sintomatologia progrediu durante oito meses, com histórico de trauma. Na mielografia, foi identificado um CAM na região lombar (L1-L2) lateralizado para a esquerda. O tratamento instituído foi a laminectomia e a durectomia. A paciente apresentou melhora dos sinais clínicos após 11 dias da realização da cirurgia. O tratamento cirúrgico obteve bons resultados para essa enfermidade. O CAM pode ocorrer em cães geriátricos ou com paraplegia de membros, assim deve ser incluído na lista de diagnóstico diferencial das mielopatias lombares compressivas.(AU)

Medullary arachnoid cyst (MAC) is a disease that occurs in humans and animals, and may cause neurological clinical signs. The origin of this disease, as well as its pathophysiology, are still unknown. It is believed that it can be congenital or acquired. No predilection for race, sex, or age has been verified. The aim of this paper is to report a lumbar MAC case in a dog at 13 years of age that caused paralysis of the pelvic limbs. At the clinical examination the patient had back pain on the lumbar region, urinary and fecal incontinence, and paraplegia on the pelvic members. The symptoms were progressing for eight months with history of trauma. In myelography a MAC in the lumbar region (L1- L2) lateralized to left was identified. For treatment laminectomy and durectomy were established. The patient showed improvement of clinical signs eleven days after surgery. The surgical treatment achieved good results for this type of disease. MAC can occur in geriatric or member paraplegia dogs, so it must be included in the differential diagnosis list of the lumbar compressive myelopathy.(AU)

AIM2 inflammasome is associated with disease severity in tegumentary leishmaniasis caused by Leishmania (V.) braziliensis.

The inflammasome is a multiprotein signalling platform involved in the pathogenesis of various inflammatory skin diseases. Herein, we investigated gene and protein expression of the inflammasome molecules AIM2 and NLRP3 in active lesions from patients with L. (V.) braziliensis-associated tegumentary leishmaniasis (TL) and correlated these findings with the clinical presentations and responses to therapy. Real-time PCR assays showed a significantly higher AIM2 gene expression in mucosal leishmaniasis (ML) compared with that in cutaneous leishmaniasis (CL). Additionally, AIM2 mRNA expression was significantly higher in lesions from poor responders than in lesions from good responders. In situ protein quantification analyses revealed greater AIM2 expression in ML lesions than in CL lesions. The percentage of AIM2-producing cells was higher in poor responders than in good responders. Although not quite significant, IL-1ß+ cells were slightly more prominent in poor responders than in good responders. Similar results were observed when patients were evaluated according to clinical form. GP63 immunostaining was identified in all samples, but no significant variation between mucosal and cutaneous lesions was observed. GP63 could be associated with reduced NLRP3 inflammasome expression in CL and ML patients. Taken together, these data demonstrate that AIM2 is an important component of the inflammasome in TL patients and is directly associated with the severity of lesions.

Implante de Port-o-Cath para quimioterapia antineoplásica em um canino: relato de caso / Implant of Port-o-Cath for antineoplastic chemotherapy in a canine: case report

O Port-o-Cath é um cateter venoso central totalmente implantável, o qual permite a infusão de quimioterápicos, hemoderivados, nutrição parenteral e coleta de sangue. Este trabalho relata o caso de um canino com um tumor no membro pélvico que foi tratado cirurgicamente e recebeu o cateter para a quimioterapia antineoplásica. A paciente possuía um sarcoma sinovial, tumor incomum em cães, que acomete normalmente as articulações e exige terapia multimodal. O tratamento foi feito em duas etapas; na primeira, o membro foi amputado e, na segunda, foi realizado o implante do Port-o-Cath e exérese de nódulo metastático. A cadela respondeu satisfatoriamente à cirurgia e realizou as sessões de quimioterapia com perfeito funcionamento do cateter, indo a óbito por outras causas 90 dias após o procedimento. O implante de Port-o-Cath para a quimioterapia é um grande aliado à terapia multimodal preconizada para o câncer, podendo o cateter ser considerado em pacientes que necessitem de medicação quimioterápica por longo período.(AU)

The Port-a-Cath is a totally implantable central venous catheter that allows the infusion of chemotherapeutic agents, blood products, parenteral nutrition, and blood sample collection. This work reports the case of a dog with a tumor in the pelvic limb that was treated surgically and received the catheter for cancer chemotherapy. The patient was diagnosed with synovial sarcoma, an uncommon tumor in dogs that usually affects the joints and requires multimodal therapy. The treatment was done in two steps, in the first the limb was amputated and in the second the Port-a-Cath implantation and the excision of the metastatic nodule were performed. The dog responded well to surgery and chemotherapy sessions held with proper functioning of the catheter, and died from other causes 90 days after the procedure. The implantation of Port-a-Cath for chemotherapy is a great ally of multimodal therapy recommended for cancer, so this catheter may be considered for patients requiring chemotherapy medication for a long period.(AU)

Agenesia renal unilateral e criptorquidismo ipsilateral em um felino: relato de caso / Unilateral renal agenesis and ipsilateral cryptorchidism in a cat: case report

A agenesia renal é uma afecção congênita rara na espécie felina, frequentemente associada a uma malformação reprodutiva. O presente trabalho relata o caso de um felino com agenesia renal unilateral associada a criptorquidismo ipsilateral, com ênfase no diagnóstico, tratamento e acompanhamento. O paciente foi conduzido ao Hospital de Clínicas Veterinárias da Universidade Federal do Rio Grande do Sul para avaliação de criptorquidismo. A agenesia renal foi um achado durante a ecografia abdominal do felino. Durante a laparotomia, foi confirmada a ausência do rim e ureter direito, hipertrofia do rim esquerdo e presença de um testículo ectópico. O paciente teve alta após a recuperação anestésica e se mantém clinicamente estável, transcorridos seis meses da cirurgia. A agenesia renal unilateral é uma condição compatível com a vida, contanto que o rim existente apresente funcionamento aceitável. Assim, sugere-se que a possibilidade de rim único em felinos criptorquidas deve ser investigada sempre que possível, tendo em vista a alta correlação entre essas malformações, e objetivando um acompanhamento da função renal do paciente ao longo da vida.(AU)

Renal agenesis is a rare disorder in feline species, commonly associated with reproductive malformation. This study aims to report the case of a cat with unilateral renal agenesis combined with ipsilateral cryptorchidism, emphasizing the diagnosis, treatment and patient follow up. The patient was taken to the Veterinary Hospital of the Federal University of Rio Grande do Sul to evaluate the cryptorchidism. The renal agenesis was an incidental finding during the abdominal ultrasound. At laparotomy, the absence of the right kidney and ureter was confirmed, hypertrophy of the left kidney and the presence of an ectopic testicle were found. The patient was discharged after recovering from anesthesia and remains clinically stable six months after surgery. The unilateral renal agenesis is a life compatible condition as long as the existing kidney has an acceptable performance. Therefore, it is suggested that the chance of a single kidney in cats whit cryptorchidism should be investigated, given the correlation between these malformations, and aiming to monitor renal function throughout the life of the patient.(AU)

Influence of the Notch system in the therapeutic response of American tegumentary leishmaniasis.

BACKGROUND: The evolution and therapeutic outcome of American tegumentary leishmaniasis (ATL) depend upon many factors, including the balance between Th1 and Th2 cytokines to control parasite multiplication and lesion extension. Other cytokines known for their role in inflammatory processes such as interleukin IL-17 or IL-18 as well as factors controlling keratinocyte differentiation and the inflammatory process in the skin, like the Notch system, could also be involved in the disease outcome. Notch receptors are a group of transmembrane proteins that regulate cell fate decisions during development and adulthood in many tissues, including keratinocyte differentiation and T-cell lineage commitment, depending on their activation by specific groups of ligands (Delta-like or Jagged). OBJECTIVES: To compare the in situ expression of Notch system proteins (receptors, ligands and transcriptional factors) and cytokines possibly involved in the disease outcome (IL-17, IL-18, IL-23 and transforming growth factor-ß) in ATL cutaneous and mucosal lesions, according to the response to therapy with N-methyl glucamine. METHODS: Cutaneous and mucosal biopsies obtained from patients prior to therapy with N-methyl glucamine were analysed by immunohistochemistry and real-time polymerase chain reaction. RESULTS: Notch receptors and Delta-like ligands were found increased in patients with ATL, particularly those with poor response to therapy or with mucosal lesions. CONCLUSIONS: The increase of Notch receptors and Delta-like ligands in patients with a poor response to treatment suggests that these patients would require a more aggressive therapeutic approach or at least a more thorough and rigorous follow-up.

Therapeutic failure in American cutaneous leishmaniasis is associated with gelatinase activity and cytokine expression.

Cutaneous lesions caused by Leishmania braziliensis infection occasionally heal spontaneously, but with antimonials therapy heal rapidly in approximately 3 weeks. However, about 15% of the cases require several courses of therapy. Matrix metalloproteinase-2 (MMP-2) and MMP-9 are gelatinases that have been implicated in other chronic cutaneous diseases and skin re-epithelialization. These enzymes are controlled by their natural inhibitors [tissue inhibitors of metalloproteinase (TIMPs)] and by some cytokines. Uncontrolled gelatinase activity may result in intense tissue degradation and, consequently, poorly healing wounds. The present study correlates gelatinase activity to therapeutic failure of cutaneous leishmaniasis (CL) lesions. Our results demonstrate an association between gelatinase activity and increased numbers of cells making interferon (IFN)-γ, interleukin (IL)-10 and transforming growth factor (TGF)-ß in lesions from poor responders. Conversely, high levels of MMP-2 mRNA and enhanced MMP-2 : TIMP-2 ratios were associated with a satisfactory response to antimonials treatment. Additionally, high gelatinolytic activity was found in the wound beds, necrotic areas in the dermis and within some granulomatous infiltrates. These results indicate the importance of gelatinase activity in the skin lesions caused by CL. Thus, we hypothesize that the immune response profile may be responsible for the gelatinase activity pattern and may ultimately influence the persistence or cure of CL lesions.

Effects of ultraviolet radiation on human cutaneous nerve fibres.

UNLABELLED: Due to an increasing number of skin diseases as a result of exposure to ultraviolet (UV) radiation, it is necessary to evaluate the effectiveness of new skin care formulations with broad-spectrum sunscreens. OBJECTIVES: This study aims to assess the status of nerve fibres in healthy human skin, to quantify effects of UV radiation on nerve endings, and to evaluate neuroprotective effects of new skin care formulations against UV exposure damage. METHODS: Samples were obtained from 34 female patients enrolled for plastic surgery and were immediately treated (10 min) with three emulsions: Cream 1, Cream 2 (placebo) and a sunscreen with sun protection factor 15 (SPF15). Control samples and those treated with the cream emulsions were exposed to UVA and UVB for 60 min. Nerve fibres were identified by immunofluorescence using a monoclonal antibody (anti-human CD56/NCAM). Cell damage was assessed by image analysis. RESULTS: Several cellular nervous structures were identified in the skin samples, including free nerve endings. UVA and UVB significantly decreased (40-60%) density of nerve endings in the control samples and those treated with placebo (Cream 2) or SPF15 (all P < 0.001). Cream 1 completely blocked effects of UV radiation on nerve endings (P > 0.05 vs. control). CONCLUSIONS: Quantification of cell damage induced by UV radiation provides useful information for identification of new skin care compounds with neuroprotective properties.

New insights to the MLL recombinome of acute leukemias.

Chromosomal rearrangements of the human MLL gene are associated with high-risk pediatric, adult and therapy-associated acute leukemias. These patients need to be identified, treated appropriately and minimal residual disease was monitored by quantitative PCR techniques. Genomic DNA was isolated from individual acute leukemia patients to identify and characterize chromosomal rearrangements involving the human MLL gene. A total of 760 MLL-rearranged biopsy samples obtained from 384 pediatric and 376 adult leukemia patients were characterized at the molecular level. The distribution of MLL breakpoints for clinical subtypes (acute lymphoblastic leukemia, acute myeloid leukemia, pediatric and adult) and fused translocation partner genes (TPGs) will be presented, including novel MLL fusion genes. Combined data of our study and recently published data revealed 104 different MLL rearrangements of which 64 TPGs are now characterized on the molecular level. Nine TPGs seem to be predominantly involved in genetic recombinations of MLL: AFF1/AF4, MLLT3/AF9, MLLT1/ENL, MLLT10/AF10, MLLT4/AF6, ELL, EPS15/AF1P, MLLT6/AF17 and SEPT6, respectively. Moreover, we describe for the first time the genetic network of reciprocal MLL gene fusions deriving from complex rearrangements.

Immune factors involved in the cervical immune response in the HIV/HPV co-infection.

AIMS: Immune factors influencing the progression of cervical intraepithelial neoplasia (CIN) to cancer remain poorly defined. This study investigates the expression of RANTES, MIP1alpha, COX1, COX2, STAT3, TGFbetaRI, IL10R, TNFalphaRII and TLR4 in the cervical immune response in HIV/HPV (human papillomavirus) co-infected women. METHODS: Cervical biopsies of 36 patients were assayed by immunohistochemistry, and the Ventana Benchmark System was used for HIV-nef detection. RESULTS: Cervices from HIV-positive patients exhibited nef in cells mainly around blood vessels, and showed a decreased expression of all the immune factors tested except IL10R and STAT3, while RANTES (5.54 cells/mm(2)) was highly expressed in comparison with controls (1.41 cells/mm(2), p = 0.028). COX1 was decreased in the HIV/HPV- (0.32 cells/mm(2), p = 0.017) and HPV-infected patients (0.21 cells/mm(2), p = 0.015) compared with controls (3.28 cells/mm(2)). CONCLUSIONS: It is suggested that RANTES in HIV/HPV co-infection may influence the development of CIN leading to progression to cervical cancer.

Liver injury and viremia in mice infected with dengue-2 virus.

The goal of this study was to test the feasibility of BALB/c mice as an experimental model in the study of dengue disease. BALB/c mice were intraperitoneal infected with DENV-2 obtained from a human patient. Histopathological analysis of infected animals revealed liver injury with viral antigens detection. In initial stages, the most prominent lesions were vacuolization and diffuse steatosis in hepatocytes. Serum levels of ALT and AST increased progressively, reaching the highest values 7 days p.i. and decreasing at the 14th day. Since levels of circulating virus were very low, viremia was analyzed in C6/36 cells. Virus presence was detected by ultrastructural analysis, confirmed by RT-PCR assays. Period of viremia was analyzed by flow cytometry with cells incubated with mouse-infected sera collected in different days, revealing peak virus levels at the 7th day p.i. All such data correlate to the development of the disease described in humans.

A comparison of horse-race, bingo, and video poker gamblers in Brazilian gambling settings.

In the present study Brazilian gamblers from different settings were compared on sociodemographic characteristics, gambling the behavior, and use of drugs. The SOGS was administered to 171 subjects at bingo (BG), video poker (VP), and horse-racing clubs (HR) of São Paulo. BG concentrated most women, VP the youngest and single gamblers, and HR the lowest income subjects. More VP than HR or BG gamblers reported taking time off work to gamble, as well as returning another day to win back lost money. They also had a higher number of scorable responses on the SOGS. The differences observed suggest that VP gamblers bear a greater risk of developing a pathological gambling pattern. The authors suggest that measures should be taken aimed at the prevention of pathological gambling, particularly among the young population of video poker gamblers.

Transplacental chemical exposure and risk of infant leukemia with MLL gene fusion.

Infant acute leukemia (IAL) frequently involves breakage and recombination of the MLL gene with one of several potential partner genes. These gene fusions arise in utero and are similar to those found in leukemias secondary to chemotherapy with inhibitors of topoisomerase II (topo-II). This has led to the hypothesis that in utero exposures to chemicals may cause IAL via an effect on topo-II. We report a pilot case-control study of IAL across different countries and ethnic groups. Cases (n = 136) were population-based in most centers. Controls (n = 266) were selected from inpatients and outpatients at hospitals serving the same populations. MLL rearrangement status was derived by Southern blot analysis, and maternal exposure data were obtained by interviews using a structured questionnaire. Apart from the use of cigarettes and alcohol, very few mothers reported exposure to known topo-II inhibitors. Significant case-control differences were apparent for ingestion of several groups of drugs, including herbal medicines and drugs classified as "DNA-damaging," and for exposure to pesticides with the last two being largely attributable, respectively, to one nonsteroidal anti-inflammatory drug, dipyrone, and mosquitocidals (including Baygon). Elevated odds ratios were observed for MLL+ve (but not MLL-ve) leukemias (2.31 for DNA-damaging drugs, P = 0.03; 5.84 for dipyrone, P = 0.001; and 9.68 for mosquitocidals, P = 0.003). Although it is unclear at present whether these particular exposures operate via an effect on topo-II, the data suggest that specific chemical exposures of the fetus during pregnancy may cause MLL gene fusions. Given the widespread use of dipyrone, Baygon, and other carbamate-based insecticides in certain settings, confirmation of these apparent associations is urgently required.

Pathological and nonpathological gamblers: a survey in gambling settings.

In this first study on gambling in Brazil, pathological and non-pathological gamblers were surveyed at three bingo clubs, one video poker club, and one horse-racing club in São Paulo. The South Oaks Gambling Screen and a questionnaire were administered to 171 subjects. When compared to nonpathological gamblers, a significantly higher proportion of pathological gamblers played cards, horse races, video poker, and dice in their lifetime. The two groups were similar with respect to socially acceptable games such as lotteries, bingo, sports, and the stock market. No significant differences were observed in drug consumption except for a higher lifetime consumption of tobacco among pathological gamblers. Only 4.9% of the gamblers sought help for gambling-related problems, suggesting that gambling is not generally perceived as a mental health problem by these subjects.

A lysine-to-arginine change found in natural alleles of the human T-cell lymphotropic/leukemia virus type 1 p12(I) protein greatly influences its stability.

The HTLV-1 singly spliced open reading frame I protein, p12(I), is highly unstable and appears to be necessary for persistent infection in rabbits. Here we demonstrate that p12(I) forms dimers through two putative leucine zipper domains and that its stability is augmented by specific proteasome inhibitors. p12(I) is ubiquitylated, and mutations of its unique carboxy-terminus lysine residue to an arginine greatly enhance its stability. Interestingly, analysis of 53 independent HTLV-1 strains revealed that the natural p12(I) alleles found in ex vivo samples of tropical spastic paraparesis-HTLV-1-associated myelopathy patients contain a Lys at position 88 in some cases, whereas arginine is consistently found at position 88 in HTLV-1 strains from all adult T-cell leukemia-lymphoma (ATLL) cases and healthy carriers studied. This apparent segregation of different alleles in tropical spastic paraparesis-HTLV-associated myelopathy and ATLL or healthy carriers may be relevant in vivo, since p12(I) binds the interleukin-2 receptor beta and gammac chains, raising the possibility that the two natural alleles might affect differently the regulation of these molecules.

Evaluation of the stability and immunogenicity of autoclaved and nonautoclaved preparations of a vaccine against American tegumentary leishmaniasis.

This study was designed to evaluate the immunogenicity of autoclaved and nonautoclaved preparations of a vaccine composed of whole antigens from killed promastigotes of Leishmania amazonensis. Leishmanin skin-test (LST)-negative volunteers were immunized with either autoclaved or nonautoclaved vaccine preparations (32 and 36 subjects, respectively) that had been maintained at 4 degrees C for one year before the onset of this trial. Immunological tests were performed two days before and 40 days after vaccination. The LST conversion rates induced by the autoclaved and nonautoclaved vaccines were significantly different: 59% and 83%, respectively. Leishmania antigen-stimulated proliferative responses of peripheral blood mononuclear cells (PBMC) were significantly higher after vaccination than before vaccination in both groups. The CD8+ subset was predominant over the CD4+ subset among the leishmania-reactive cells after vaccination in both groups. The production of IFN-gamma by the leishmania antigen-stimulated PBMC was significantly higher after vaccination than before vaccination in the group receiving the nonautoclaved vaccine but not in the autoclaved vaccine group. IL-2 was found both before and after vaccination with no differences between its levels in these time points in either group. IL-4 was not detected for either group during the study period.

Changes in metabolism of inorganic polyphosphate in rat tissues and human cells during development and apoptosis.

Age-dependent studies show that the amount of inorganic polyphosphate in rat brain strongly increases after birth. Maximal levels were found in 12-months old animals. Thereafter, the concentration of total polyphosphate decreases to about 50%. This decrease in the concentration of total polyphosphate is due to a decrease in the amount of insoluble, long-chain polyphosphates. The amount of soluble, long-chain polyphosphates does not change significantly in the course of ageing. In rat embryos and newborns, mainly soluble polyphosphates could be detected. In rat liver, the age-dependent changes are less pronounced. The changes in polyphosphate level are accompanied by changes in exopolyphosphatase activity, which degrades the polymers to orthophosphate; highest enzyme activities were found when the polyphosphate level was low. Induction of apoptosis in the human leukemic cell line HL-60 by actinomycin D results in degradation of long polyphosphate chains. The total polyphosphate content does not change significantly in apoptotic cells.

A novel method for determination of inorganic polyphosphates using the fluorescent dye fura-2.

A method for determining inorganic polyphosphate, which is based on the Mn(2+)-induced quenching of the fluorescence of the calcium indicator fura-2, is described. The effect of Mn2+ ions on fura-2 fluorescence is gradually abolished in the presence of increasing concentrations of polyphosphate; this allows the quantification both of synthetic polyphosphates and of the naturally occurring polymer isolated from tissues or cells. The described method has some advantages compared to conventional procedures for detection of polyphosphates based on the metachromatic effect on toluidine blue. It can be applied for the determination of pyrophosphate, tripolyphosphate and other short-chain polyphosphates not detectable by toluidine blue and it can be used for measurement both of pyrophosphatase and exopolyphosphatase activity.

T-cell responsiveness of American cutaneous leishmaniasis patients to purified Leishmania pifanoi amastigote antigens and Leishmania braziliensis promastigote antigens: immunologic patterns associated with cure.

Patients suffering from American cutaneous leishmaniasis were studied before therapy (active lesion) and at the end of therapy (cured patients). Assays of lymphocyte proliferative responses of peripheral blood mononuclear cells induced in vitro by Leishmania braziliensis promastigote antigens (Lb) or by three proteins (A-2/P-2, P-4, and P-8) derived from Leishmania pifanoi amastigotes were performed. Antigen-stimulated cells were harvested for CD4 and CD8 phenotype analysis and the levels of gamma interferon (IFN-gamma), interleukin 2 (IL-2) and interleukin 4 (IL-4) produced were also determined. Results show two different patterns of Lb-induced T cell responses: (a) predominance of responding CD4+ cells and mixed type 1 and type 2 cytokine production (IFN-gamma, IL-2, and IL-4) during the active disease, (b) similar proportions of responding CD4+ and CD8+ cells and type 1 cytokine production (presence of IFN-gamma and IL-2 and very low IL-4) at the end of therapy (healed lesions). Thus, this last pattern is probably associated with a beneficial T cell response. The A-2/P-2 amastigote cysteine proteinase provided only marginal (s.i. approximately or = 2.5) T cell stimulation in 25% of patients studied; in contrast, the L. pifanoi P-4 and P-8 amastigote antigens induced significant stimulation (s.i. approximately or = 5) in approximately 50% of the patients. In comparison to Lb-stimulated cultures, lower proliferative responses of T lymphocytes to P-4 or P-8 were observed. However, the P-4- or P-8-stimulated cultures had similar percentages of reactive CD4+ and CD8+ cells, as well as type 1 cytokines (presence of IFN-gamma and IL-2, and low levels or absence of IL-4) in the supernatants both before and at the end of therapy. The consistent induction of apparently beneficial T cell responses by the P-4 and P-8 amastigote glycoproteins points to the possibility that these molecules be considered as candidates for future defined vaccines against leishmaniasis.