Immune cells from patients with psoriasis are defective in inducing indoleamine 2,3-dioxygenase expression in response to inflammatory stimuli.

BACKGROUND: Indoleamine 2,3-dioxygenase (IDO) is an inducible enzyme that suppresses the immune response. The role of IDO as a negative regulator of inflammatory responses has been documented in several experimental autoimmune diseases. OBJECTIVES: To explore the regulation of IDO by immune cells in psoriasis and its relation with disease severity. METHODS: The expression and activity of IDO were assessed by reverse-transcriptase polymerase chain reaction, flow cytometry and high-performance liquid chromatography in peripheral blood of patients with moderate-to-severe plaque-type psoriasis. The ability of immune cells to express IDO in response to inflammatory stimuli was studied. The functional role of IDO expression was evaluated in a regulatory T cell (Treg) differentiation assay, using cocultures of immature monocyte-derived dendritic cells with autologous peripheral CD4+ T cells. RESULTS: Analysis of the kynurenine-to-tryptophan ratio in serum samples indicated higher IDO activity in patients with psoriasis than in healthy controls. However, correlation studies showed lower IDO activity in those patients with higher Psoriasis Area and Severity Index (PASI). Although myeloid dendritic cells from patients with psoriasis expressed higher levels of IDO than those from healthy controls, these cells did not upregulate IDO in response to a combination of tumour necrosis factor-α, interleukin (IL)-1ß and IL-6 cytokines. The defective expression of IDO correlated with PASI. Immature monocyte-derived dendritic cells from patients with psoriasis also expressed low levels of IDO and induced CD4+ Treg differentiation poorly. CONCLUSIONS: Immune cells from patients with psoriasis have a defect in upregulating IDO in response to inflammation associated with the severity of psoriasis.

Reduced expression of galectin-1 and galectin-9 by leucocytes in asthma patients.

Accumulating evidence shows that galectins play roles in the initiation and resolution phases of inflammatory responses by promoting anti- or proinflammatory effects. This study investigated the presence of three members of the galectin family (galectin-1, -3 and -9) in induced sputum samples of asthma patients, as well as their possible implication in the immunopathogenesis of human asthma. Levels of interleukin (IL)-5, IL-13, and galectins were determined in leucocytes isolated from induced sputum samples by reverse transcription-polymerase chain reaction (RT-PCR) immunofluorescence and flow cytometry. High levels of IL-5 and IL-13 mRNA were detected in sputum cells from asthma patients. In parallel, immunoregulatory proteins galectin-1 and galectin-9 showed a reduced expression on macrophages from sputum samples compared with cells from healthy donors. In-vitro immunoassays showed that galectin-1 and galectin-9, but not galectin-3, are able to induce the production of IL-10 by peripheral blood mononuclear cells from healthy donors. These findings indicate that macrophages from sputum samples of asthma patients express low levels of galectin-1 and galectin-9, favouring the exacerbated immune response observed in this disease.

[Jejunal mucormycosis in a patient with Hodgkin's lymphoma]. / Mucormicosis yeyunal en paciente con linfoma de Hodgkin.

We report a case of intestinal mucormycosis in a 46-year-old male diagnosed with classical Hodgkin's disease, IV-B stage. During the first phase of chemotherapy he had a massive digestive bleeding event secondary to a jejunal ulcer, and zygomicosis mucor-type was diagnosed by endoscopic biopsy. The patient was treated with antifungal drugs and surgical resection of the intestine involved. At surgery a double covered perforation of the jejunum was seen. Pathological examination confirmed the previous diagnosis. After one year of follow-up the patient is doing well, and his lymphoma is on remission. To our best knowledge this is the second case of intestinal mucormycosis in a patient with Hodgkin's lymphoma reported in the medical literature.

Consumo de energía y nutrientes, características socioeconómicas, pobreza y área de residencia de mujeres peruanas en edad fértil / Energy and nutrient consumption, socio-economic characteristics, poverty and living places fertile peruvian women

Objetivo: Determinar la relación entre el consumo de energía y nutrientes de mujeres peruanas en edad fértil, sus características socioeconómicas, pobreza y área de residencia. Materiales y métodos: La información fue obtenida de la Encuesta Nacional de Consumo de Alimentos en Mujeres en Edad Fértil y Niños entre 12 y 35 meses en el año 2003. La muestra fue de 2909 mujeres entre 15 a 49 años. El consumo de energía y nutrientes se obtuvo por el método de encuesta de recordatorio de 24 horas. Se incluyó información del área de residencia, características y servicios básicos del hogar. Se agrupó a la población de acuerdo con el método de necesidades básicas insatisfechas (NBI). Se estableció como consumo deficiente una adecuación por debajo de 50 por ciento. Se aplicaron pruebas estadísticas de Chi-cuadrado, Mann Whitney y Kruskall Wallis. Resultados: El 67,6 por ciento de las mujeres vivía en hogares pobres, la pobreza afecta mayormente a las mujeres del área rural. Más de 20 por ciento de mujeres presentan deficiencias en el consumo de energía y nutrientes. Conforme se incrementan las NBI las adecuaciones del consumo van disminuyendo, a excepción de carbohidratos y hierro. Los carbohidratos contribuyen en mayor porcentaje a la energía total de la dieta de las mujeres pobres. Conclusiones: Las condiciones de pobreza en que se encuentran las mujeres influyen sobre su consumo de energía y nutrientes disminuyéndolo, lo cual traería serias consecuencias sobre su estado de salud y nutrición y el de sus niños.

Objectives: To determine the relationship between energy and nutrient consumption in fertile Peruvian women, as well as their socioeconomic characteristics, poverty condition, and living places. Materials and methods: Data were obtained from the National Food Consumption Survey for Fertile Women and 12 to 35 Month Old Children for 2003. The sample consisted in 2909 women between 15 to 49 years old. Energy and nutrient consumption information was obtained using a 24-hour reminder survey. Collected information included living places, as well as households characteristics and basic services. The population was grouped according to the unmet basic needs method. A deficient consumption was established as a less than 50 per cent adequation. Statistical tests used were chi-square, Mann-Whitney, and Kruskall-Wallis. Results: 67.6 per cent of surveyed women lived in poor households, and poverty mainly affects women from rural areas. More than 20 of women had deficiencies in energy and nutrient consumption. As long as unmet basic needs increase, adequate consumption is reduced, except carbohydrate and iron consumption. Carbohydrates are the major contributors for total energy intake in poor women. Conclusions: Poverty in women negatively influences energy and nutrient consumption, which may lead to serious consequences on their health status and their childrenÆs health condition as well.

Shiga toxin 1: damage to DNA in vitro.

Shiga toxins share with plant ribosome-inactivating proteins the same enzymatic mechanism of action: the removal of a specific adenine from 28S RNA when acting on ribosomes and the removal of multiple adenines when acting on DNA in vitro. The activity on DNA, only recently reported, is particularly evident, and has been studied mostly at acidic pH. For the in vitro activity, on both ribosomes and DNA, Shiga toxins require activation by trypsin, urea and dithiothreitol which release the enzymatically active A(1) fragment. Activation by the classical procedure leaves large amounts of urea and DTT which interfere in the DNA depurination assay and completely abolish depurination at physiological pH. A consistent release of [3H]adenine from DNA at neutral pH is instead observed when the toxin is activated in vitro by an improved method which removes most of the drastic reagents required for proteolytic cleavage and reduction. Damage to single-stranded DNA by Shiga toxin 1 (Stx1) primarily involves depurination. A spontaneous DNA breakdown appears in fact only after extensive base removal, a behavior similar to that observed with uracil-DNA glycosylase, a simple glycosylase devoid of lyase activity. NaCl inhibits the activity of Stx1, probably by minimizing the sliding distance traveled by the enzyme along DNA in search of its target sites and promoting dissociation of the substrate-enzyme complex.

[An aggressive inguinal (parafunicular) angiomyxoma in a male patient]. / Angiomixoma agresivo inguinal (parafunicular) en paciente varón.

OBJECTIVE: To report a case of aggressive inguinal angiomyxoma in a male patient. METHODS: An 82-year-old male patient presented with a well-defined, 6 cm. parafunicular mass in the right groin. The mass was located adjacent to the spermatic cord and had been noted 8 years earlier. Patient evaluation included CT, ultrasound and immunohistochemical studies. RESULTS: The CT and US findings suggested lymph node enlargement. Microscopic analysis showed a myxoid tumor with partially infiltrating margins, vascular channels of small-sized vessels with thick walls occasionally with hyalinization and spindle-shaped or stellate mesenchymal cells with ill-defined margins without atypia or mitosis that were positive for vimentin and negative for actin, desmin, keratins, CD34 and protein S-100. No tumor recurrence or metastasis has been observed at 26-months' follow-up. CONCLUSIONS: To our knowledge, this is one of the few cases of inguinal angiomyxoma in male patients; 16 have been reported to date. This neoplasm appears to originate from pelvic soft tissue fibroblasts.

[Inhalation administration of nitric oxide during selective pulmonary ventilation decreases the intrapulmonary shunt]. / La administración inhalatoria de óxido nítrico durante la ventilación pulmonar selectiva disminuye el shunt intrapulmonar.

OBJECTIVE: To assess the effects of inhaled nitric oxide (NO) on oxygenation and hemodynamics in patients undergoing lung resection surgery during one-lung ventilation (OPV). PATIENTS AND METHODS: Prospective study of 16 patients aged 62 +/- 10 years scheduled for chest surgery under combined general and epidural anesthesia. During ventilation of only one lung, NO was administered for 15 minutes. Arterial blood and mixed venous blood samples were taken for analysis of blood gases and the calculation of intrapulmonary shunt. Pulmonary and systemic hemodynamic variables were also recorded using a Swan-Ganz catheter at three times: baseline (ventilation of both lungs), OLV, and with OLV plus NO (OLV NO). RESULTS: The most relevant data consisted of a significant decrease in shunt after start of NO inhalation in comparison with the level during OLV (31.1 +/- 0.5% versus 36 +/- 0.6%; p < 0.05). Arterial oxygen pressure decreased significantly during OLV and increased after start of NO (118.9 +/- 53.6 versus 155.4 +/- 78.5 mmHg; p < 0.05). Mean pulmonary artery pressure, pulmonary and systemic vascular resistances, and cardiac index did not change with inhalation of NO. CONCLUSIONS: Inhalational administration of NO during OLV significantly improves arterial oxygenation and decreases intrapulmonary shunt during OLV, without causing hemodynamic or systemic effects.

[Inflammatory pseudotumor of the spleen. An old concept with many questions]. / Pseudotumor inflamatorio de bazo. Un viejo concepto con múltiples interrogantes.

Inflammatory pseudotumor of spleen is an infrequent benign condition. It is difficult to differentiate, on a clinical and radiological basis, from haematologic neoplasms, granulomatous diseases as sarcoidosis and splenic hamartoma. Sometimes can be an incidental finding. Two women, aged 72 years, are presented. On the first case the sympthons mi micked a malignant disease. The second one was an incidental finding in a routine study for cholecystitis. Histological and immunohistochemical study showed a polymorphic cellular population including plasma cell, lymphoid cells, histiocytes, eosinophils and spindle cells, showing a reactive benign character. Plasma cells presented light chains polyclonality. Lymphoid cells were mature and with T inmunophenotype. Spindle cells were focally positive for muscle spe-cific actin and vimentine. In the first case, ultraestructural study showed myofibroblast morphology on the stromal spindle cells. Like many other authors have already postulated, immunohistochemical and ultraestructural findings would corroborate the mesenchymal reactive and benign nature of this type of lesions.

Parotid metastasis from renal clear cell adenocarcinoma. An unusual site for metastasis.

Metastasis to the parotid region is rare, and originates primarily from head and neck squamous cell carcinoma and melanoma of the skin. Renal clear cell adenocarcinoma has considerable metastatic potential and the parotid gland is one possible destination. Histopathologic study is important to differentiate this entity from primary clear cell parotid neoplasm.

Pyrrolidine dithiocarbamate inhibits the production of interleukin-6, interleukin-8, and granulocyte-macrophage colony-stimulating factor by human endothelial cells in response to inflammatory mediators: modulation of NF-kappa B and AP-1 transcription factors activity.

Endothelial cells (EC) play a key role in the inflammatory response, both by the production of proinflammatory cytokines and by their interaction with leukocytes. Molecular genetic analysis has demonstrated that functional NF-kappa B sites are involved in the transcription of interleukin-6 (IL-6), IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) genes in response to inflammatory mediators. Thus, we have explored the effect of two inhibitors of the NF-kappa B activation, pyrrolidine dithiocarbamate (PDTC) and N-acetylcysteine (NAC), on the production of these cytokines by EC. Both PDTC and NAC inhibited, in a dose-dependent manner, the synthesis of IL-6, IL-8, and GM-CSF induced by tumor necrosis factor (TNF)-alpha or bacterial lipopolysaccharides (LPS) in human umbilical vein endothelial cells (HUVEC). PDTC appeared to prevent IL-6, IL-8, and GM-CSF gene transcription, as it blocked the induction of specific mRNA by TNF-alpha or LPS. The TNF-alpha mediated transcriptional activation of a chloramphenicol acetyltransferase (CAT) plasmid containing three copies of the -72 kappa B binding site from the IL-6 promoter was abrogated by PDTC. According to transfection experiments, electrophoretic mobility shift assays (EMSA) demonstrated that the antioxidant prevented the induction of NF-kappa B DNA-binding activity by TNF-alpha. Under the same conditions, PDTC by itself or in combination with TNF-alpha, enhanced the DNA-binding activity of AP-1, as well as c-fos and c-jun mRNA levels. Altogether, these results indicate that the antioxidant PDTC specifically inhibits the transcription of IL-6, IL-8, and GM-CSF genes through the inhibition of the NF-kappa B activation, while increasing the expression of AP-1. Our data make evident the antiinflammatory and immunoregulatory potential of the pharmacological inhibition of the NF-kappa B activation. In addition, PDTC and related molecules may be a useful tool to explore the expression of genes involved in the inflammatory response.

Transcriptional up-regulation of intracellular adhesion molecule-1 in human endothelial cells by the antioxidant pyrrolidine dithiocarbamate involves the activation of activating protein-1.

The redox status of the cell plays an essential role in regulating signal transduction, transcription factor activity, and expression of cell surface molecules. In this study, we show that pyrrolidine dithiocarbamate (PDTC), a potent antioxidant agent, upregulated the cell surface expression of intercellular adhesion molecule-1 (ICAM-1) in human endothelial cells (EC). Further analysis of PDTC-mediated ICAM-1 up-regulation revealed that PDTC increased ICAM-1 mRNA levels and augmented its gene promoter activity. Transfection experiments in EC with reporter constructs harboring nested deletion fragments of the ICAM-1 promoter indicated the presence of a functional PDTC-responsive region located between positions -136 to -353 of the promoter. Gel retardation assays together with supershift analysis revealed that PDTC induced the binding of c-fos and c-jun to a consensus activating protein-1 (AP-1) binding site located at position -284. PDTC alone or in combination with TNF-alpha enhanced AP-1-dependent transactivation in HUVEC, as determined by DNA binding assays. The functional implication of AP-1 in the transcription of the ICAM-1 gene was further demonstrated by cotransfection experiments in which a c-jun expression vector induced the promoter activity of the PDTC-responsive element of the ICAM-1 promoter. Taken together, these results indicate that the antioxidant PDTC induces transcriptional activation of ICAM-1 and that this induction is mediated at least in part by the transcription factor AP-1. This mechanism might be operative in pathologic conditions in which a redox imbalance plays a key role, such as ischemia/reperfusion injury or arteriosclerosis.

[Angiocentric immunoproliferative lesions: the terminology and the clinical picture]. / Lesiones angiocéntricas inmunoproliferativas: terminología y cuadros clínicos.

Two diagnosed clinical cases of T-cell angiocentric lymphoma are presented, different in their clinical aspects, evolution and treatment. Both represent different spectrums within the entity of angiocentric immunoproliferative lesions. The medical literature is reviewed and emphasis is placed on the different illness which include a wide range from less to more highly malignant within the same entity.

Characterization of the CD11a (alpha L, LFA-1 alpha) integrin gene promoter.

Human lymphocyte function-associated antigen-1 (LFA-1, CD11a/CD18, alpha L/beta 2) is a cell surface heterodimer, which, together with Mac-1 (CD11b/CD18, alpha M/beta 2) and p150,95 (CD11c/CD18, alpha X/beta 2), constitutes the leukocyte integrin beta 2 (CD18) subfamily. LFA-1 is the only integrin expressed on all leukocyte lineages and functions both as a key adhesion receptor in immune and inflammatory processes and as a signal-transducing molecule. To elucidate the molecular basis for the leukocyte-restricted expression of LFA-1, the promoter region of the CD11a gene has been isolated and functionally characterized. The 5' region of the CD11a gene exhibits a similar exon/intron organization as the CD11b, CD11c, and VLA-2 alpha genes but is different from that of the genes encoding VLA-4 alpha, VLA-5 alpha, and gpIIb. Several tightly clustered transcription initiation sites have been identified on the CD11a gene, with the major site resembling the "initiator" sequence. Transient expression of CD11a promoter-based reporter gene constructs in both LFA1+ and LFA1- cell lines demonstrated that the fragment spanning from -880 to +83 is involved in the tissue-specific expression of LFA-1. Functional analysis of different fragments within the -880/+83 fragment suggested the presence of negative regulatory elements between -880 and -226 and demonstrated that the proximal region of the CD11a gene promoter exhibits tissue-specific activity.

Characterization of the p150,95 leukocyte integrin alpha subunit (CD11c) gene promoter. Identification of cis-acting elements.

The leukocyte integrin p150,95 (CD11c/CD18) is involved in a number of cell-cell and cell-extracellular matrix interactions and mediates signal transduction into the cytoplasm. p150,95 is expressed on cells of the myeloid lineage as well as on certain activated T and B lymphocytes, and its expression is regulated during cell activation and differentiation. Since CD18 is expressed on all leukocyte lineages, the restricted expression of p150,95 must be controlled at the level of CD11c gene transcription. To understand the mechanisms that direct the constitutive and regulated leukocyte expression of p150,95 we have structurally characterized the CD11c promoter region and initiated its functional dissection. The CD11c promoter lacks TATA- and CCAAT-boxes, directs the synthesis of transcripts with heterogeneous 5'-ends, and contains an initiator-like sequence at the major transcription initiation site. Several putative binding sequences for ubiquitous (Sp1, AP-1, AP-2, and NF-kB) and leukocyte-specific (PU.1) transcription factors have been identified in the proximal region of the CD11c promoter which may participate in the regulation of the expression of p150,95. Transient expression of CD11c-based reporter gene constructs indicates that the CD11c promoter dictates the tissue-specific expression of p150,95 and that sequences contained within 160 base pairs 5' from the major transcriptional start site are involved in the tissue-specific and regulated expression of p150,95. DNase I protection analysis on the promoter region spanning from -160 to +40 revealed four regions of DNA-protein interactions (FPI-FPIV), two of which (FPII and FPIV) correlate with the cell type-specific and regulated expression of the CD11c gene.

A single mRNA encodes the alpha 150 and alpha 80/70 forms of the alpha subunit of VLA4.

VLA4 is a cell surface heterodimer (alpha 4 beta 1, CD49d/CD29) which belongs to the integrin family and is involved in cell-extracellular matrix interactions, as well as in intercellular adhesion. Unlike other integrin alpha subunits, the alpha subunit of VLA4 (alpha 4) can appear as a 150-kDa polypeptide (alpha 150), or cleaved into two non-covalently associated polypeptides (alpha 70-80). The relative proportion of each form is highly variable among different cells and is dependent on the state of cellular activation. The alpha 4 cleavage site has recently been shown to occur between residues Arg558-Ser559. We report the isolation of genomic clones encoding the alpha 4 subunit, the location of the cleavage site-encoding nucleotides to a specific exon and the determination of the exon-intron organization around the cleavage site. Comparison with the gpIIb (CD41) and the alpha x (p150,95 alpha, CD11c) genes revealed a similar genomic structure in this region, with exons of similar length separated by introns of identical phase. The structure of the alpha 4 mRNA in cells expressing the alternative forms of alpha 4 has been analyzed by means of reverse transcription-polymerase chain reaction. Our results indicate that the exon encoding the cleavage site is present in alpha 4 mRNA molecules from cells expressing either the alpha 150 or the alpha 70-80 form on the cell surface. Moreover, the structure of the alpha 4 mRNA around the cleavage site does not change during the switch towards the alpha 70-80 form that takes place upon lymphocyte activation. Therefore, both forms of alpha 4 arise from a common mRNA, the alpha 150 form contains the cleavage sequence and the alpha 70-80 form must be generated by a post-translational proteolytic event.

Endometrioid adenosarcoma of the bladder arising from endometriosis.

Vesical endometriosis is an uncommon entity characterized by the deposition of benign, hyperplastic endometrial tissue in the bladder. To date, only about 160 cases have been reported in the world literature. More uncommon, however, are case reports of extrauterine malignancies arising from pre-existing endometriosis. We report a case of a large endometrial adenosarcoma in a patient who had undergone total abdominal hysterectomy and bilateral salpingo-oophorectomy 10 years previously for endometriosis. To our knowledge this malignancy has never been reported previously to involve the bladder. We review the pathological findings, possible mode of entry into the bladder and current concepts in its treatment.