Reduced Meloidogyne ethiopica Parasitism in Vitis vinifera Grafted on Six Resistant Rootstocks Under Field and Greenhouse Conditions.

Meloidogyne ethiopica, an aggressive nematode, causes significant economic losses to Vitis crops. Rootstocks can successfully manage phytoparasitic nematodes. However, no studies exist on M. ethiopica-resistant rootstocks under field conditions. This study assessed the resistance of six Vitis rootstocks to M. ethiopica under field and greenhouse conditions. The number of galls and eggs in vine roots, quantity of second stage juveniles and males in 250 ml of soil, root weight, and shoot weight were determined for the Harmony, SO4, 101-14 MG, 110R, 3309C, and Kober 5BB rootstocks, and the own-root Chardonnay variety as a control. In the field, Harmony, SO4, 101-14 MG, Kober 5BB, and 110R were highly resistant to nematode parasitism and reproduction. In turn, 3309C was the least resistant rootstock. In the greenhouse, all rootstocks similarly limited M. ethiopica attack and reproduction. In both conditions, Chardonnay was the most susceptible vine to nematode attack, with high quantities of galls, eggs, and nematode reproduction. In conclusion, most of the evaluated rootstocks reduced M. ethiopica parasitism. Harmony, SO4, 101-14 MG, 110R, and Kober 5BB rootstocks are options for vineyard use, with final selection dependent on winegrower requirements.

Low-Volume High-Intensity Interval Training as a Therapy for Type 2 Diabetes.

Our purpose was to investigate the effects of low-volume, high-intensity interval training (HIT) on cardiometabolic risk and exercise capacity in women with type 2 diabetes mellitus (T2DM). Sedentary overweight/obese T2DM women (age=44.5±1.8 years; BMI=30.5±0.6 kg/m(2)) were randomly assigned to a tri-weekly running-based HIT program (n=13) or non-exercise control follow-up (CON; n=10). Glycemic control, lipid and blood pressure levels, endurance performance, and anthropometry were measured before and after the follow-up (16 weeks) in both groups. Medication intake was also assessed throughout the follow-up. Improvements (P<0.05) on fasting glucose (14.3±1.4%), HbA1c (12.8±1.1%), systolic blood pressure (3.7±0.5 mmHg), HDL-cholesterol (21.1±2.8%), triglycerides (17.7±2.8%), endurance performance (9.8±1.0%), body weight (2.2±0.3%), BMI (2.1±0.3%), waist circumference (4.0±0.5%) and subcutaneous fat (18.6±1.4%) were found after HIT intervention. Patients of HIT group also showed reductions in daily dosage of antihyperglycemic and antihypertensive medication during follow-up. No changes were found in any variable of CON group. The HIT-induced improvements occurred with a weekly time commitment 56-25% lower than the minimal recommended in current guidelines. These findings suggest that low-volume HIT may be a time-efficient intervention to treat T2DM women.

PstS-1, the 38-kDa Mycobacterium tuberculosis glycoprotein, is an adhesin, which binds the macrophage mannose receptor and promotes phagocytosis.

Mycobacterium tuberculosis, the primary causative agent of tuberculosis, infects macrophages and transforms the hostile intracellular environment into a permissive niche. M. tuberculosis infects macrophages using a variety of microbial ligand/cell receptor systems. In this study, binding assays with biotin-labelled mycobacterial cell wall proteins revealed five Concanavalin A-reactive proteins that bind macrophages. Among these proteins, we identified PstS-1, a 38-kDa M. tuberculosis mannosylated glycolipoprotein, and characterized it as an adhesin. Inhibition assays with mannan and immunoprecipitation demonstrated that PstS-1 binds the mannose receptor. We purified PstS-1 to 95.9% purity using ion exchange chromatography. The presence of mannose in purified PstS-1 was demonstrated by Concanavalin A interaction, which was abolished in the presence of sodium m-periodate and α-D-mannosidase. Gas chromatography revealed that purified PstS-1 contained 1% of carbohydrates by weight, which was mainly mannose. Finally, we used fluorescent microbeads coated with purified PstS-1 in phagocytosis assays and discovered that microbead uptake was inhibited by the pre-incubation of cells with GlcNAc, mannan and α-methyl mannoside. The interaction of PstS-1 coated beads with the mannose receptor was confirmed by confocal colocalization studies that showed high Pearson and Manders's colocalization coefficients. Our findings contribute to a better understanding of the strategies M. tuberculosis uses to infect host cells, the critical first step in the pathogenesis of tuberculosis.

Ancient photosynthetic eukaryote biofilms in an Atacama Desert coastal cave.

Caves offer a stable and protected environment from harsh and changing outside prevailing conditions. Hence, they represent an interesting habitat for studying life in extreme environments. Here, we report the presence of a member of the ancient eukaryote red algae Cyanidium group in a coastal cave of the hyperarid Atacama Desert. This microorganism was found to form a seemingly monospecific biofilm growing under extremely low photon flux levels. Our work suggests that this species, Cyanidium sp. Atacama, is a new member of a recently proposed novel monophyletic lineage of mesophilic "cave" Cyanidium sp., distinct from the remaining three other lineages which are all thermo-acidophilic. The cave described in this work may represent an evolutionary island for life in the midst of the Atacama Desert.

Mycobacterium tuberculosis 38-kDa lipoprotein is apoptogenic for human monocyte-derived macrophages.

Mycobacterium tuberculosis is the main aetiologic agent of tuberculosis, a disease of great concern in less-developed regions. Apoptosis is a conspicuous event in macrophages infected in vitro with mycobacteria, a phenomenon also observed in vivo in granulomas of patients with tuberculosis. To determine its significance, it is important to define the mycobacterial moieties involved and how they cause apoptosis. Here we show that the 38-kDa lipoprotein induces macrophage caspase-dependent apoptosis involving TNF-alpha and FasL and, interestingly, with the upregulation of cell-death receptors TNFR1, TNFR2 and Fas. A role for the Toll-like receptor 2 was also demonstrated. In conclusion, the ability to induce apoptosis of host cells is another property of the 38-kDa lipoprotein, a molecule that has focused attention for being an immunodominant antigen that participates in phosphate transport.

[Study on the venoms of the principal venomous snakes from French Guiana and the neutralization]. / Etude des venins des principaux serpents venimeux de Guyane française et de leur neutralisation.

We studied some biochemical, toxic and immunological characteristics of the venoms of Bothrops atrox, Bothrops brazili and Lachesis muta, Viperidae responsible for most of the bites of venomous snakes in French Guiana. Chromatographic (HPLC) and electrophoretical profiles (SDS-PAGE), lethal, hemorrhagic, defibrinogenating, coagulant, thrombin like, proteolytic, fibrino(geno)lytic and phospholipase activities were studied. In addition, the neutralization of some toxic activities conferred by four antivenins was compared. The chromatographic and electrophoretic profiles were different for the three venoms, showing differences between Bothrops and L. muta venoms. In general, bothropic venoms showed the highest toxic and enzymatic activities, while the venom of L. muta showed the lowest lethal, hemorrhagic and coagulant activities. The enzymes of bothropic venoms responsible for gelatinolytic activity were around 50-90 kDa. All the venoms were able to hydrolyze a and beta chains of the fibrinogen, showing different patterns of degradation. Although all the antivenoms tested were effective to various degrees in neutralizing the venom of B. brazili and B. atrox, neutralization of L. muta venom was significantly better achieved using the antivenom including this venom in its immunogenic mixture. For the neutralization of L. muta venom, homologous or polyvalent antivenoms that include the "bushmaster" venom in their immunogenic mixture should be preferred.

Coexpression of NRAMP1, iNOS, and nitrotyrosine in bovine tuberculosis.

In murine models the inducible nitric oxide synthase (iNOS) and the natural resistance associated macrophage protein (NRAMP1) play major roles in host defense against mycobacteria. iNOS regulates nitric oxide (NO) production, which is noxious for ingested mycobacteria, and NRAMP1 displays pleiotropic antimicrobial effects, including upregulation of iNOS expression. Little is known about the role of these molecules in bovine tuberculosis (TB). In this work we demonstrate by Western blot a high expression of NRAMP1 in peripheral blood mononuclear cells (PBMCs), alveolar macrophages (obtained by bronchioalveolar lavage), and lymph node granulomas from 8 Holstein-Freisian cattle with autopsy-proven bovine TB. Immunohistochemistry revealed the abundant expression of NRAMP1 and iNOS in lymph node and lung granulomas. Immunoreactivity was abundant in the cytoplasm of many epithelioid macrophages and multinucleated giant cells of the Langhans type. A striking accumulation of nitrotyrosine (NT), an indicator of iNOS activity and local NO production, was observed in granuloma cells, particularly in multinucleated Langhans cells. This study shows that the expression of NRAMP1 and iNOS is costimulated in granulomas, which are protective T-cell reactions against mycobacteria.

Biocompatibility and toxicological studies of carbon nanotubes doped with nitrogen.

In this report, we compare the toxicological effects between pure carbon multiwalled nanotubes (MWNTs) and N-doped multiwalled carbon (CNx) nanotubes. Different doses of tubes were administered in various ways to mice: nasal, oral, intratracheal, and intraperitoneal. We have found that when MWNTs were injected into the mice's trachea, the mice could die by dyspnea depending on the MWNTs doses. However, CNx nanotubes never caused the death of any mouse. We always found that CNx nanotubes were far more tolerated by the mice when compared to MWNTs. Extremely high concentrations of CNx nanotubes administrated directly into the mice's trachea only induced granulomatous inflammatory responses. Importantly, all other routes of administration did not induce signs of distress or tissue changes on any treated mouse. We therefore believe that CNx nanotubes are less harmful than MWNTs or SWNTs and might be more advantageous for bioapplications.

An IgG antibody response to the antigen 85 complex is associated with good outcome in Mexican Totonaca Indians with pulmonary tuberculosis.

SETTING: It is generally accepted that antibodies do not protect against Mycobacterium tuberculosis infection, as this role relies upon T-cell reactivity. Hence, most studies on antimycobacterial antibodies have been aimed at developing serologic tests, and few explore their role in disease pathogenesis. OBJECTIVE: To determine the IgG antimycobacterial antibody response of 55 Mexican Totonaca Indians with pulmonary tuberculosis and its correlation with some features of the disease. DESIGN: Study of the profile of antigen recognition by immunoblot and ELISA with isolated antigen 85 complex (Ag85) and whole culture filtrate proteins. Correlation of immunoblot and ELISA results with BCG vaccination, tuberculin reactivity, extent of the disease, clinical setting, and response to treatment. RESULTS: On immunoblot, band reactivity was very poor and the most frequently recognized antigen was the 30-32 kDa, antigen 85 complex (45.8% of serum samples). ELISA with this antigen showed a sensitivity of 72% and a specificity of 100%. Positive antibody titers to Ag85 were observed in 79.4% of patients with non-cavitary tuberculosis (P = 0.012) and in 95.8% of patients who were cured with anti-tuberculosis chemotherapy (P = 0.0001). By contrast, an antibody response to whole culture filtrate antigens had no correlation with the presence of cavitations or with prognosis. CONCLUSIONS: Our data show that an antibody response to Ag85, aside from having great potential to develop a serologic test for tuberculosis, was associated with a positive outcome in a cohort of tuberculous Mexican Indians.

High-level expression of NRAMP1 in peripheral blood cells and tuberculous granulomas from Mycobacterium bovis-infected bovines.

By Western blotting, we demonstrate high-level expression of NRAMP1 proteins in peripheral blood cells and granulomas of Mycobacterium bovis-infected bovines. Immunohistochemistry of granulomatous lesions showed heavily labeled epithelioid macrophages and Langhans cells. These data suggest that M. bovis infection enhances NRAMP1 expression and that active tuberculosis can occur despite this response.

Binding and activation of human plasminogen by Mycobacterium tuberculosis.

The first evidence of the interaction of Mycobacterium tuberculosis with the plasminogen system is herein reported. By FACScan analysis and affinity blotting, lysine-dependent binding of plasminogen to M. tuberculosis was demonstrated. The binding molecules were 30-, 60-, and 66-kDa proteins present in cell wall and soluble protein extracts. The activation of plasminogen, which occurred only in presence of fibrin and was not inhibited by the host serpin, alpha(2)-antiplasmin, was also demonstrated.

Co-stimulatory signals increase the reactivity of gammadelta T cells towards mycobacterial antigens.

Although it has been shown that gammadelta T lymphocytes are able to react with different cell-associated or soluble antigens, the immune repertoire of these cells appears to be skewed to the recognition of mycobacterial antigens. We have studied the number and reactivity of gammadelta T cells towards several mycobacterial antigens in patients with tuberculosis and leprosy, as well as their healthy contacts and control individuals. We found an increased number of Vdelta2+ cells in healthy contacts (PPD+ and lepromin+) and tuberculoid leprosy patients. The gammadelta T cells from lepromatous leprosy showed a decreased response to all antigens tested, but some of these patients exhibited a significant response to the 30-kD glycoprotein of Mycobacterium tuberculosis. Interestingly, the reactivity of gammadelta T cells against mycobacterial antigens was significantly increased by costimulatory signals generated through CD7, LFA-1, CD50 and CD69 in all groups. However, signalling through CD69 did not enhance the responsiveness of gammadelta lymphocytes from lepromatous patients. On the other hand, the in vitro blockade of IL-10 with a specific antibody enhanced the cell proliferation of gammadelta lymphocytes from lepromatous leprosy patients, whereas exogenous IL-10 had an opposite effect in most individuals studied. These results suggest the potential role of different cell membrane receptors in the regulation of gammadelta T cell proliferation induced by mycobacteria, as well as the possible involvement of IL-10 in this phenomenon.

Rapid screening test for tuberculosis using a 38-kDa antigen from Mycobacterium tuberculosis.

A screening test for the diagnosis of tuberculosis by immunodot (IDt) is described, using an antigen of Mycobacterium tuberculosis, namely, a 38-kDa glycoprotein which has shown great specificity in previous serologic analyses. The test was used to examine 28 sera from patients with lung tuberculosis. Of these, 85% were positive by micro-ELISA and by the IDt test herein described. Control sera from healthy subjects (n = 20) gave negative results for ELISA and for IDt, which indicates that the screening test is highly specific. The test is easy to handle and requires no equipment and is therefore particularly useful for field studies.

Mycobacterium fortuitum glycolipids for the serodiagnosis of pulmonary tuberculosis.

Glycolipids belonging to the family of acylated trehaloses were isolated from Mycobacterium fortuitum, a rapidly growing mycobacterial species, and tested in the serologic diagnosis of human pulmonary tuberculosis by enzyme-linked immunosorbent assay. Di- and tri-O-acylated trehaloses from M. fortuitum reacted with serum antibodies of patients with pulmonary tuberculosis at higher titers than did with sera from healthy donors. With both glycolipids, the sensitivity of the test was above 0.80 at a chosen specificity of 0.98. Individuals with treated tuberculosis showed lower antibody titers compared with their initial reactivities. These data show that M. fortuitum could be used as a surrogate source of antigens for tuberculosis serodiagnosis.

T-cell lung granulomas induced by sepharose-coupled Mycobacterium tuberculosis protein antigens: immunosuppressive phenomena reversed with cyclophosphamide and indomethacin.

We induced lung granulomas in BALB/c mice by intratracheal instillation of Sepharose beads coated with a Mycobacterium tuberculosis protein extract. Granulomas composed of macrophages and lymphocytes were induced. The granulomatous reaction reached its peak 3-7 days after challenge and lasted for approximately 1 month. Immunolabelling of tissue sections and bronchial washings revealed that granulomas were predominantly composed of T lymphocytes with the cytotoxic-suppressor phenotype (CD8+). Granulomas were associated with a significant decrease in anti-mycobacterial immunity manifested by a drop in delayed-type hypersensitivity reactions and antibody titres. The immunosuppressive phenomena were abolished with cyclophosphamide or indomethacin. Control granulomas induced with methylated bovine serum albumin (BSA) were smaller and composed by similar numbers of CD4+ and CD8+ cells. BSA granulomas did not alter antibody titres but they decreased delayed-type hypersensitivity to BSA which was restored to normal with indomethacin but not with cyclophosphamide. Our findings show that mycobacterial proteins anchored to Sepharose beads are granulomatogenic and that they preferentially recruit CD8+ cells which, together with locally produced prostaglandins, down-modulate cell-mediated and humoral immunity to mycobacterial antigens.

Antigenic and structural similarities between Mycobacterium tuberculosis 50- to 55-kilodalton and Mycobacterium bovis BCG 45- to 47-kilodalton antigens.

The relationship between Mycobacterium tuberculosis 50- to 55-kDa protein and Mycobacterium bovis BCG 45- to 47-kDa antigen was examined by using immunological and biochemical criteria. Reciprocal cross-reactivity with a rabbit polyclonal antiserum against the M. bovis BCG protein and with a monoclonal antibody raised against the M. tuberculosis antigen was observed. The epitope recognized by this antibody was apparently present only in proteins of M. tuberculosis and M. bovis BCG among the 11 mycobacterial species tested. The amino-terminal sequences and total amino acid contents of these proteins showed strong similarities. Both antigens are glycoproteins as assessed by binding of concanavalin A, labeling of carbohydrate moieties with biotin-hydrazide, and digestion of carbohydrates with jack bean alpha-D-mannosidase, which produced a reduction of the molecular weights of the proteins and totally eliminated concanavalin A binding. Both M. tuberculosis and M. bovis BCG proteins are secreted, since they were found mainly in the culture medium. Analysis of M. tuberculosis 50- to 55-kDa antigen by two-dimensional gel electrophoresis showed at least seven different components, as previously described for the M. bovis BCG antigen. Solid-phase immunoassays showed that the purified M. tuberculosis 50- to 55-kDa protein was recognized by serum specimens from 70% of individuals with pulmonary tuberculosis from a total of 77 Mexican patients examined.

Mammary 5'deiodinase (5'D) during the breeding cycle of the rat: indirect evidence that 5'D type I is specific to the alveolar epithelium.

The present study analyses the activity of 5'deiodinases type I and II in mammary gland during the breeding and estrous cycle of the rat, and includes indirect evidence that 5'D-I is present only in the alveolar epithelium. Data show that the mammary gland exhibits 5'D-II activity throughout the developmental period and its activity varies along the allometric growth of the gland. 5'D-I is detected during the differentiation stages of the alveolar epithelium (puberty, late pregnancy) and its activity rises significantly (10-fold) 24 h after delivery. Data also show that 5'D-I activity is not present in the fat pads of the gland. These findings suggest that during its differentiation and functional stages, the mammary gland requires an elevated and compartmentalized production of T3.