Identification of phospholipid species affected by miltefosine action in Leishmania donovani cultures using LC-ELSD, LC-ESI/MS, and multivariate data analysis.

Leishmaniasis is a widespread parasitic disease principally treated by intravenous drugs. Hexadecylphosphocholine (miltefosine) has recently proved its efficacy by oral route. Although its mechanism of action has been investigated, and principally relies on perturbations of the metabolism of lipids and especially phospholipids, further studies need to be conducted to detect precisely which metabolic pathways are impacted. For this purpose, the present work proposes a complete lipidomic study focused on membrane phospholipids of clones of Leishmania donovani non-treated (NT), treated (T) and resistant (R) to miltefosine. Firstly, a separation of phospholipids in normal phase high-performance liquid chromatography (NP-HPLC) was coupled to a mass spectrometer (MS) equipped with an electrospray (ESI) ion source, and response was compared to evaporative light scattering detection (ELSD). Secondly, a quantification of phospholipid classes was performed using NP-HPLC/ESI/MS on NT, T and R clones of L. donovani. Thirdly, full-scan acquisitions of analyzed samples were compared using orthogonal signal correction-partial least square-discriminant analysis (OSC-PLS-DA) to highlight phospholipid molecular species of interest between the three types of clones. Structural determination of the most relevant species has finally been performed using tandem mass spectrometry. A first hypothesis on the effect of miltefosine on lipid metabolic pathways is then proposed.

Controlling coherence using the internal structure of hard pi pulses.

The tiny difference between hard pi pulses and their delta-function approximation can be exploited to control coherence. Variants on the magic echo that work despite a large spread in resonance offsets are demonstrated using the zeroth- and first-order average Hamiltonian terms, for 13C NMR in 60C. The 29Si NMR linewidth of silicon has been reduced by a factor of about 70,00 using this approach, which also has potential applications in magnetic resonance microscopy and imaging of solids.

Bartonella native valve endocarditis: the first brazilian case alive and well

Bartonella is an important cause of blood culture-negative endocarditis in recent studies. Seroprevalence studies in the States of Minas Gerais and Rio de Janeiro have shown Bartonella IgG positivity around 14 percent in healthy adults and 40 percent in HIV seropositive adults, respectively. A case report of a 46-year-old white male with moderate aortic regurgitation (AR) due to rheumatic heart disease (RHD), admitted due to worsening heart failure, is presented. Clinical features were apyrexia, anemia, polyclonal hypergammaglobulinemia, hematuria and splenomegaly. He was submitted to surgery due to worsening AR. Histopathology of the excised valve showed active bacterial endocarditis and underlying RHD. Routine blood cultures were negative. Indirect immunofluorescence (IFI) assays for Coxiella burnetii were non-reactive. Bartonella henselae IgG titer was 1:4096 prior to antibiotics and 1:512 14 months after treatment. History of close contact with a young cat during the months preceding his admission was elicited.

Generating unexpected spin echoes in dipolar solids with pi pulses.

NMR spin echo measurements of 13C in C60, 89Y in Y2O3, and 29Si in silicon are shown to defy conventional expectations when more than one pi pulse is used. Multiple pi-pulse echo trains may either freeze out or accelerate the decay of the signal, depending on the pi-pulse phase. Average Hamiltonian theory, combined with exact quantum calculations, reveals an intrinsic cause for these coherent phenomena: the dipolar coupling has a many-body effect during any real, finite pulse.

Bartonella native valve endocarditis: the first Brazilian case alive and well.

Bartonella is an important cause of blood culture-negative endocarditis in recent studies. Seroprevalence studies in the States of Minas Gerais and Rio de Janeiro have shown Bartonella IgG positivity around 14% in healthy adults and 40% in HIV seropositive adults, respectively. A case report of a 46-year-old white male with moderate aortic regurgitation (AR) due to rheumatic heart disease (RHD), admitted due to worsening heart failure, is presented. Clinical features were apyrexia, anemia, polyclonal hypergammaglobulinemia, hematuria and splenomegaly. He was submitted to surgery due to worsening AR. Histopathology of the excised valve showed active bacterial endocarditis and underlying RHD. Routine blood cultures were negative. Indirect immunofluorescence (IFI) assays for Coxiella burnetii were non-reactive. Bartonella henselae IgG titer was 1:4096 prior to antibiotics and 1:512 14 months after treatment. History of close contact with a young cat during the months preceding his admission was elicited.

Very extended shapes in the A--110 region.

High-angular-momentum states in 108Cd were populated via the (64)Ni((48)Ca,4n) reaction at a beam energy of 207 MeV. Gamma rays were detected using the Gammasphere array. A rotational band has been observed with a dynamic moment of inertia and deduced lower limit of the quadrupole moment suggesting a major-to-minor axis ratio larger than 1.8:1, placing it among the most deformed structures identified in any nucleus, to date.

rst and its paralogue kirre act redundantly during embryonic muscle development in Drosophila.

The polynucleate myotubes of vertebrates and invertebrates form by fusion of myoblasts. We report the involvement of the Drosophila melanogaster Roughest (Rst) protein as a new membrane-spanning component in this process. Rst is strongly expressed in mesodermal tissues during embryogenesis, but rst null mutants display only subtle embryonic phenotypes. Evidence is presented that this is due to functional redundancy between Rst and its paralogue Kirre. Both are highly related single-pass transmembrane proteins with five extracellular immunoglobulin domains and three conserved motifs in the intracellular domain. The expression patterns of kirre and rst overlap during embryonic development in muscle founder cells. Simultaneous deletion of both genes causes an almost complete failure of fusion between muscle founder cells and fusion-competent myoblasts. This defect can be rescued by one copy of either gene. Moreover, Rst, like Kirre is a myoblast attractant.

Ubiquitous overexpression of a transgene encoding the extracellular portion of the Drosophila roughest-irregular chiasm C protein induces early embryonic lethality.

The cell adhesion molecule Rst-irreC is a transmembrane glycoprotein of the immunoglobulin superfamily involved in several important developmental processes in Drosophila, including axonal pathfinding in the optic lobe and programmed cell death and pigment cell differentiation in the pupal retina. As an initial step towards the "in vivo" functional analysis of this protein we have generated transgenic fly stocks carrying a truncated cDNA construct encoding only the extracellular domain of Rst-IrreC under the transcriptional control of the heat shock inducible promoter hsp70. We show that heat-shocking embryos bearing the transgene during the first 8hs of development lead to a 3-4 fold reduction in their viability compared to wild type controls. The embryonic lethality can already be produced by applying the heat pulse in the first 3hs of embryonic development, does not seem to be suppressed in the absence of wildtype product and is progressively reduced as the heat treatment is applied later in embryogenesis. These results are compatible with the hypothesis of the lethal phenotype being primarily due to heterophilic interactions between Rst-IrreC extracellular domain and an yet unknown ligand.

[Depression in the elderly and the role of family dynamics]. / La depresión del anciano y el papel de la dinámica familiar.

OBJECTIVE: The objective was to find the relationship between the lived experience within the family and depression in the elderly. DESIGN: The study had a crossover and descriptive design. SETTING: Carranque and Campanillas Health Districts, Málaga. PATIENTS: The target population was the over 65's. The sample consisted of 97 people attended at 2 health centres, one rural and one urban. MEASUREMENTS AND MAIN RESULTS: The main study variable was the results on the Yesavage Geriatric Depression Scale (GDS), which were contrasted through multiple linear regression with the variables of age, sex, marital status, tobacco dependency, number of people living together, presence of the children in the home (concretely, daughters), family income, ownership of the home, social support network, and Apgar Family II. The results showed that the GDS correlated only with the results of the Apgar Family II (p = 0.00003). CONCLUSIONS: When tackling the depression of elderly people in primary care, it is absolutely vital to look at the family context, as this is a factor strongly linked to the depression.

La depresión del anciano y el papel de la dinámica familiar / Depression in the elderly and the role of family dynamics

Objetivo. El objetivo es conocer la relación entre la vivencia de la dinámica familiar y la depresión en el anciano. Diseño. El diseño del estudio es descriptivo, transversal. Emplazamiento. Zona Básica de Salud de Carranque y Zona Básica de Salud de Campanillas, ambas de Málaga. Pacientes. La población diana es la población mayor de 65 años. La muestra está constituida por 97 sujetos atendidos en 2 centros de salud, uno rural y otro urbano. Mediciones y resultados principales. La principal variable de estudio es el resultado de la Escala de Depresión Geriátrica (GDS) de Yesavage, cuyos resultados se correlacionan mediante regresión lineal múltiple con las variables edad, sexo, estado civil, tabaquismo, número de convivientes, presencia de hijos en el domicilio (y concretamente, de hijas), ingresos familiares, titularidad de propiedad sobre el domicilio, red de apoyo social y Apgar Familiar II. Los resultados obtenidos muestran que el GDS se correlaciona únicamente con los resultados del Apgar Familiar II (p = 0,00003). Conclusiones. En el abordaje de la depresión del anciano en atención primaria resulta de gran importancia considerar el ámbito familiar por ser éste un factor fuertemente asociado a aquélla (AU)

Retrospective study on dengue fatal cases.

Immunohistochemical procedure (avidin biotin peroxidase complex) was applied in formalin-fixed and paraffin-embedded tissues obtained from 5 fatal cases of dengue infection associated with encephalopathy. Dengue virus antigen was demonstrated in the cytoplasm of phagocytic mononuclear cells from liver, spleen, and lung. Moreover, dengue viral antigens were here, to our knowledge, first demonstrated in the central nervous system (CNS) and numerous immunolabelled cells were found in brain sections from 3 cases. Extended immunohistochemical studies carried out in 1 case showed virus-positive cells mostly located within Virchow Robin space of medium size and small veins, infiltrating the white and grey matter, and often situated close to neurons displaying apparent cytopathic features. Furthermore, immunostaining for CD68 antigens demonstrated that most CD68+ macrophages and dengue antigen-positive cells share similar morphology and localization, suggesting a unique identity for at least part of these cells. Since in dengue fever, virus replicates mostly in cells of macrophage lineage, our results seem to indicate that infiltration of virus-infected macrophages could be one of the pathways by which viruses enter the brain in dengue encephalitis. Whether bone marrow-derived infected macrophages and viral-free particles induce CSN lesions through immune, metabolic, and/or direct viral-induced mechanisms will be essential to better understand the pathogenesis and provide new therapeutic strategies for dengue-associated encephalitis. As the evidence of tissue damage was nonspecific, the detection of virus antigen by immunoperoxidase technique appeared to be highly reliable for dengue diagnosis.

[Cardiac changes in children with AIDS]. / Alterações cardíacas em crianças com SIDA.

PURPOSE: To describe the cardiac findings in children with AIDS, the follow-up with treatment and the correlation between clinical and pathological features. METHODS: We studied prospectively 25 children with clinical-laboratorial diagnosis of AIDS, ages between 3 months and 11 years, even those without cardiac symptoms. We classified the signs of AIDS following that of CDC-Atlanta 1994. Eight children died and it was done necropsy in six with macro and microscopic examinations. RESULTS: Fifteen cases had already complications of AIDS and were classified as C2 and C3, 5 as B2, 3 B1 and 2 as A1. Beside the symptoms related to the disease and infections we found signs of congestive heart failure III and IV (NYHA) in 5 children, pericardial effusion in 5, (one of them had cardiac tamponade). In the electrocardiogram (EKG), 8 children had repolarization abnormalities. In 1st echocardiogram (ECHO) we found some features of dilated myocardiopathy in 8 (6 were asymptomatic) 5 of those had pericardial effusion, one child had huge amount of pericardial liquid and also increased measures of intraventricular septum and posterior wall of left ventricle by ECHO and confirmed by necropsy examination. All cases of diagnosis of myocardiopathy received treatment with captopril and 4 also received furosemide and digoxina. Even under oral treatment three of them had deterioration of heart failure but after combined anti-retroviral drugs they showed better cardiac functions. From the six necropsied cases, two had increased heart weight without myocarditis, one had toxoplasma pancarditis and other one had fibrocalcic vasculopathy. CONCLUSION: We found several assymptomatic cases already with decreased ventricular function. Some worsened of heart failure even under apropriate treatment and showed better cardiac index after combined anti-retroviral drugs.

[Rheumatic heart disease and infective endocarditis in a patient with acquired immunodeficiency syndrome]. / Cardiopatia reumática e endocardite infecciosa em paciente com síndrome de imunodeficiência adquirida.

A 36-old-woman was admitted with an infectious syndrome, respiratory insufficiency and vasculitis. There was a history of chronic intravenous drug abuse, sexual promiscuity and rheumatic heart disease. She had HIV positive tests. The vasculitis and heart failure worsened and the patient died of stroke. At autopsy it was found histologic evidence of AIDS, rheumatic heart disease with Aschoff nodes, infective endocarditis with cerebral abscesses and thalamic infarction.

When neurobiology and genetics meet: the study of visual system development in Drosophila melanogaster.

The visual system of Drosophila melanogaster is an exceptionally well suited model to study the molecular and genetic mechanisms underlying inter- and intracellular signalling during nervous system development. This review highlights some of the recent advances in this field, that show not only the power of the combined neurobiological and genetical approaches in shedding new light into long standing questions regarding nervous system development but also uncover a striking evolutionary conservation, at the molecular level, of the basic developmental pathways and signal transduction cascades underlying metazoan ontogenesis.

Restricted expression of the irreC-rst protein is required for normal axonal projections of columnar visual neurons.

The 104 kDa irreC-rst protein, a member of the immunoglobulin superfamily, mediates homophilic adhesion in cell cultures. In larval optic chiasms, the protein is found on recently formed axon bundles, not on older ones. In developing visual neuropils, it is present in all columnar domains of specific layers. The number of irreC-rst-positive neuropil stratifications increases until the midpupal stage. Immunoreactivity fades thereafter. The functional importance of the restricted expression pattern is demonstrated by the severe projection errors of axons in the first and second optic chiasms in loss of function mutants and in transformants that express the irreC-rst protein globally. Epigenesis of the phenotypes can be explained partially on the bases of homophilic irreC-rst interactions.

[Clinicopathologic correlation in 50 cases of acquired immunodeficiency syndrome. Retrospective study]. / Correlação clínicopatológica de 50 casos de síndrome de imunodeficência adquirida. Estudo retrospectivo.

PURPOSE: To analyze myocardial abnormalities in patients of acquired immunodeficiency syndrome with clinical and pathological correlation. METHODS: We selected 50 cases, retrospectively, age ranged from 3 months to 40 years, all of them had myocardial changes and the data of clinical records fulfilled our protocol. Cases of others cardiac diseases were not included. RESULTS: The pathological findings were: myocarditis in 33 (11 had severe myocarditis) and degenerative hystological lesions in 17. The etiologic agents detected were: Toxoplasma in 11, Cryptococcus in 7 and Cytomegalovirus in 3. In 12 cases we could not find any agent. In 15 cases occurred others lesions: endocarditis, pericarditis and sarcoma of Kaposi. It was noted tachycardia in 15 cases, decrease of heart sounds in 12, arterial hypotension in seven, systolic murmur in 8, galop rhythm in 7, pericardial friction rub in 3, arrhythmia in 2. Four patients had congestive heart failure. The EKG showed sinus tachycardia in 18, ST and T changes in 10, low voltage in 5, ST segment elevation in 5 and extrasystoles in 3 cases. The echocardiogram findings were: pericardial effusion in 9 cases and 9 had ventricular dysfunction. CONCLUSION: The cardiac lesions were very important even in patients without clinical signals. We need others prospective studies with viral identification trying to detect specific lesions of HIV.

The irregular chiasm C-roughest locus of Drosophila, which affects axonal projections and programmed cell death, encodes a novel immunoglobulin-like protein.

The axonal projection mutations irregular chiasm C of Drosophila melanogaster comap and genetically interact with the roughest locus, which is required for programmed cell death in the developing retina. We cloned the genomic region in 3C5 by transposon tagging and identified a single transcription unit that produces a major, spatially and temporally regulated mRNA species of approximately 5.0 kb. Postembryonic expression is strong in the developing optic lobe and in the eye imaginal disc. The gene encodes a transmembrane protein of 764 amino acids with five extracellular immunoglobulin-like domains and similarity to the chicken axonal surface glycoprotein DM-GRASP/SC1/BEN. Both known irreC alleles reduce the level of transcription, whereas the roughestCT mutation disrupts the intracellular domain of the protein.

A prospective, placebo-controlled, randomized trial of intravenous streptokinase and angioplasty versus lone angioplasty therapy of acute myocardial infarction.

BACKGROUND: The value of routine administration of intravenous thrombolytic agents during percutaneous transluminal coronary angioplasty (PTCA) therapy of acute myocardial infarction (MI) has not been determined. Therefore, we prospectively randomized 122 patients with evolving MI to PTCA therapy with or without adjunctive intravenous streptokinase therapy. METHODS AND RESULTS: Patients with ECG ST segment elevation who presented within 4 hours of symptom onset, had no contraindication to thrombolytic therapy, and were not in cardiogenic shock were enrolled. They were treated immediately with intravenous heparin (10,000 units) and oral aspirin (325 mg) and randomized to treatment with placebo or streptokinase (1.5 M units) administered intravenously over 30 minutes. Patients then were taken immediately to the catheterization laboratory, and those with suitable coronary anatomy underwent immediate PTCA. Subsequent clinical course, serial radionuclide ventriculography, and 6-month repeat angiography were analyzed. A total of 106 patients were treated with PTCA. Use of PTCA was similar for placebo (92%) and streptokinase (83%) groups. Angioplasty was successful in 95% of patients, with no difference in placebo (93%) and streptokinase (98%) groups. Serial radionuclide ventriculography demonstrated no difference in 24-hour (52 +/- 12% versus 50 +/- 12%) or 6-week (51 +/- 12% versus 51 +/- 13%) ejection fraction values for placebo and streptokinase groups, respectively. Contrast ventriculography demonstrated improvement in immediate (54 +/- 12%) versus 6-month (60 +/- 15%, p < 0.05) values for the overall group. No differences in 6-month values were present (58 +/- 15% versus 62 +/- 15%, p = NS) for placebo and streptokinase groups, respectively. Coronary angiography was performed in 75% of the 90 patients eligible for restudy. Arterial patency was 87% at 6 months, and coronary restenosis was present in 38% of patients. No differences in chronic patency or restenosis were detected for the two treatment groups. Although adjunctive intravenous streptokinase therapy did not improve outcome, it did complicate the hospital course. Hospitalization was longer (9.3 +/- 5.0 versus 7.7 +/- 4.4 days, p = 0.046) and more costly ($25,191 +/- 15,368 versus $19,643 +/- 7,250, p < 0.02). Transfusion rate was higher (39% versus 8%, p = 0.0001) and need for emergency coronary bypass surgery was greater (10.3% versus 1.6%, p = 0.03) for the streptokinase-treated patients. CONCLUSIONS: Adjunctive intravenous streptokinase therapy does not enhance early preservation of ventricular function, improve arterial patency rates, or lower restenosis rates after PTCA therapy of acute MI. Hospital course is longer, more expensive, and more complicated. For these reasons, PTCA therapy of acute MI should not be routinely performed with adjunctive intravenous streptokinase therapy.

Outcome of patients with acute myocardial infarction who are ineligible for thrombolytic therapy.

OBJECTIVE: To determine what proportion of patients with acute myocardial infarction are not eligible for thrombolytic therapy and to assess their natural history. DESIGN: Retrospective chart review. SETTING: A large community-based hospital. PATIENTS: All patients with acute myocardial infarction hospitalized during a 27-month period. MEASUREMENTS: Of 1471 patients with acute myocardial infarction, 230 (16%) received thrombolytic therapy according to the protocol and an additional 97 (7%) received nonprotocol thrombolytic therapy, primary coronary balloon angioplasty, or both because of contraindications. The other 1144 patients (78%) did not receive reperfusion therapy. MAIN RESULTS: The patients who did not receive thrombolytic therapy were older, more likely to be women, and more likely to have a history of hypertension, previous myocardial infarction, or chronic angina (all comparisons, P less than 0.002). An average of 1.9 reasons for exclusion were identified per patient among the ineligible patients. Mortality was fivefold higher among ineligible patients (19%; Cl, 16% to 21%) than among protocol-treated patients (4%; Cl, 1% to 6%) (P less than 0.001). In-hospital mortality rates for excluded patients were 28% (Cl, 23% to 32%) in elderly patients (age, greater than 76 years; n = 396); 29% (Cl, 23% to 35%) in patients with stroke or bleeding risk (n = 209); 17% (Cl, 14% to 20%) in patients with delayed presentation (greater than 4 hours after the onset of chest pain; [n = 599]); 14% (Cl, 11% to 16%) in patients with an ineligible electrocardiogram (ECG) (n = 673); and 26% (Cl, 21% to 32%) in patients with a miscellaneous reason for exclusion (n = 243). Independent predictors of increased mortality were: age greater than 76 years, stroke or other bleeding risk, ineligible ECG, or the presence of two or more exclusion criteria. CONCLUSIONS: Thrombolytic therapy is currently used in the United States for only a minority of patients with acute myocardial infarction: those who have low-risk prognostic characteristics.

Dengue: neuropathological findings in 5 fatal cases from Brazil.

Neuropathological examination of 5 patients with dengue who died of shock in Rio de Janeiro during an outbreak in summer 1987, showed nonspecific lesions (edema, vascular congestion, hemorrhagic foci and perivascular lymphocytic infiltrates). In one case with delayed marked neurological symptoms, several foci of perivenous demyelination were observed. Neurological manifestations are various and not uncommon in dengue, but their anatomical substratum is not known. An immunopathological mechanism has been postulated in some cases but has never been demonstrated morphologically. The perivenous leukoencephalitis observed in one of our cases could represent the morphological substratum of such an immunological mechanism.