MMP1, MMP9, and COX2 expressions in promonocytes are induced by breast cancer cells and correlate with collagen degradation, transformation-like morphological changes in MCF-10A acini, and tumor aggressiveness.

Tumor-associated immune cells often lack immune effector activities, and instead they present protumoral functions. To understand how tumors promote this immunological switch, invasive and noninvasive breast cancer cell (BRC) lines were cocultured with a promonocytic cell line in a Matrigel-based 3D system. We hypothesized that if communication exists between tumor and immune cells, coculturing would result in augmented expression of genes associated with tumor malignancy. Upregulation of proteases MMP1 and MMP9 and inflammatory COX2 genes was found likely in response to soluble factors. Interestingly, changes were more apparent in promonocytes and correlated with the aggressiveness of the BRC line. Increased gene expression was confirmed by collagen degradation assays and immunocytochemistry of prostaglandin 2, a product of COX2 activity. Untransformed MCF-10A cells were then used as a sensor of soluble factors with transformation-like capabilities, finding that acini formed in the presence of supernatants of the highly aggressive BRC/promonocyte cocultures often exhibited total loss of the normal architecture. These data support that tumor cells can modify immune cell gene expression and tumor aggressiveness may importantly reside in this capacity. Modeling interactions in the tumor stroma will allow the identification of genes useful as cancer prognostic markers and therapy targets.

Interleukin-4 and RANTES expression in maturing eosinophils derived from human cord blood CD34+ progenitors.

Eosinophils elaborate a number of proinflammatory mediators, including immunoregulatory cytokines and chemokines. Interleukin (IL)-4 and RANTES are important cytokines that have previously been shown to be expressed by mature eosinophils. We hypothesized that de novo synthesis of IL-4 and RANTES occurs in nascent eosinophils, leading to storage of newly produced proteins in crystalloid granule-like structures. Cytokine mRNA and protein expression were examined in cultured eosinophil colonies, which were derived from purified cord blood CD34+ cells and generated in semisolid media (methylcellulose) in the presence of recombinant human (rh)IL-3 and rhIL-5. Cytokine mRNA profiles were analysed by the reverse transcription-polymerase chain reaction (RT-PCR) to determine transcription of IL-4 and RANTES in cells on days 0, 7, 14, 21 and 28 of culture. The expression of translated cytokine products and granule major basic protein (MBP) was confirmed, from day 23 onwards, for colonies cultured in semisolid media, by immunofluorescent labelling and confocal laser-scanning microscopy (CLSM). We found that mRNA sequences encoding IL-4 and RANTES were expressed in freshly prepared, non-differentiated CD34+ cells. Furthermore, RANTES mRNA localized to carbol chromotrope 2R-positive colony cells, as assessed using in situ RT-PCR on day 21 of culture in semisolid media, and was found to gradually decrease (relative to beta2-microglobulin) in rhIL-3- and rhIL-5-treated colony cells (comprising > 90% eosinophil-like cells) up to day 28. Immunoreactivity for IL-4 and RANTES co-localized with MBP in maturing colony eosinophils on day 23 of culture in semisolid media, as judged by CLSM. These results suggest that synthesis and storage of immunoregulatory cytokines, essential for processes associated with adaptive immunity, occurs in nascent eosinophils during their growth and differentiation.

Immunofluorescence analysis of cytokine and granule protein expression during eosinophil maturation from cord blood-derived CD34 progenitors.

BACKGROUND: In allergic inflammation and asthma, eosinophils are major effector cells. They have been shown to synthesize at least 23 cytokines, some of which are stored intracellularly in their unique crystalloid granules together with cationic granule protein. Little is known about the synthesis and storage of cytokines relative to cationic granule proteins in maturing eosinophils during eosinophilopoiesis. OBJECTIVE: Our purpose was to analyze the expression of eosinophil-derived mediators, major basic protein (MBP), eosinophil cationic protein (ECP), IL-6, and RANTES, during early stages of eosinophil maturation in CD34(+) cell-derived colonies. METHODS: Purified human cord blood CD34(+) cells were grown in methylcellulose cultures in the presence of recombinant human IL-3 and IL-5. By confocal laser scanning microscopy, the coexpression of eosinophil granular proteins MBP and ECP was determined concurrently with IL-6 and RANTES during eosinophil maturation on days 16, 19, 23, and 28 of culture. RESULTS: Immunoreactivity against MBP, ECP, IL-6, and RANTES was not detectable in freshly purified CD34(+) cells. Maturing eosinophils (>95%) exhibited positive immunostaining for all these proteins between days 16 and 28 of culture. At early stages of culture, discrete immunostaining was observed around the periphery but not in the center of granular structures. By day 28 cultured eosinophil-like cells showed evidence of the acquisition of crystalloid granule-like structures, analogous to those observed in mature peripheral blood eosinophils. CONCLUSIONS: Eosinophils express and store cytokines simultaneously with cationic granule proteins during the process of maturation. We propose that the storage of cytokines during the development of eosinophils is an early event and it may be integral to inflammatory responses involving these cells. The results of this study suggest a potential immunoregulatory function for maturing eosinophils.

Deformidad en coxa vara en la disóstosis cleidocraneal / Coxa vara deformity in cleidocranial dysostosis

La disostosis cleidocraneal es una rara displasia ósea generalizada. La coxa vara o disminución anormal del ángulo de inclinación del cuello femoral está frecuentemente presente en la disostosis cleidocraneal. Se presenta un nuevo caso y se discute la revisión de la bibliografía (AU)

Replenishment of RANTES mRNA expression in activated eosinophils from atopic asthmatics.

Eosinophils have been shown to express the gene encoding regulated upon activation, normal T-cell expressed and secreted (RANTES), a potent eosinophilotactic chemokine. RANTES protein expression in eosinophils has previously been shown to be up-regulated by a number of agonists, including complement-dependent factors (C3b/iC3b) and interferon-gamma (IFN-gamma). We hypothesized that gene expression of RANTES is regulated in these cells by eosinophil-specific agonists. We analysed RANTES mRNA expression by reverse-transcription polymerase chain reaction (RT-PCR) in human peripheral blood eosinophils obtained from mild atopic asthmatics following stimulation over time. In resting eosinophils, a low level of RANTES mRNA was found to be constitutively expressed in all the atopic donors tested in this study (n = 6). Following stimulation with C3b/iC3b (serum-coated surfaces), eosinophils released measurable levels of RANTES, while sustained transcript expression was detected for up to 24 hr of stimulation. In contrast, IFN-gamma (5 ng/ml) transiently and significantly (P<0.05, n = 3) depleted relative amounts of RANTES PCR product (compared with beta2-microglobulin) after 1-4 hr of stimulation. RANTES transcript was again detectable after 24 hr of IFN-gamma incubation, suggesting that the pool of RANTES mRNA had been replenished. Other eosinophil-active cytokines, interleukin-3 (IL-3), IL-4, IL-5 and granulocyte-macrophage colony-stimulating factor, did not appear to modulate RANTES mRNA expression after 1 hr of incubation. The effect of IFN-gamma on RANTES mRNA was reversed by cycloheximide, suggesting that IFN-gamma may act by increasing the rate of translation of RANTES mRNA. These findings indicate that IFN-gamma may induce a rapid and transient effect on the translation and replenishment of RANTES mRNA in eosinophils. This novel observation supports the notion that eosinophils have the potential to replenish their stored and released bioactive proteins.

Movilidad anormal de hombros / Abnormal motility of the shoulders

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Rapid mobilization of intracellularly stored RANTES in response to interferon-gamma in human eosinophils.

The CC chemokine RANTES is synthesized, stored, and upregulated in response to interferon-gamma (IFN-gamma) in human peripheral blood eosinophils. In this report, we propose that RANTES is rapidly mobilized from eosinophil crystalloid granules during agonist-induced degranulation. We stimulated purified eosinophils (>99%) from atopic asthmatics with 500 U/mL IFN-gamma to analyze the kinetics of mobilization and release of RANTES (0 to 240 minutes). We used subcellular fractionation, immunogold analysis, two-color confocal laser scanning microscopy (CLSM), and enzyme-linked immunosorbent assay (ELISA) to trace the movement of eosinophil-derived RANTES from intracellular stores to release. RANTES was rapidly mobilized (10 minutes) and released after 120 minutes of stimulation (80 +/- 15 pg/mL per 2 x 10(6) cells). RANTES appeared to be stored in at least two intracellular compartments: the matrix of crystalloid granules, detected by major basic protein and eosinophil peroxidase activities, and a specialized small secretory vesicle present in light membrane fractions. The extragranular RANTES was mobilized more rapidly than that of crystalloid granules during IFN-gamma stimulation. This effect was not observed in eosinophils treated with IFN-alpha, interleukin-3 (IL-3), IL-5, granulocyte-macrophage colony-stimulating factor (GM-CSF), or genistein followed by IFN-gamma. Our findings suggest that RANTES may be mobilized and released by piecemeal degranulation upon stimulation, involving transport through a putative pool of small secretory vesicles.

Effects of interferon-gamma on mobilization and release of eosinophil-derived RANTES.

Eosinophils synthesize and release a number of cytokines and chemokines, including RANTES, a potent chemoattractant particularly for memory T cells and eosinophils. Long-term (>12 h) incubation with interferon-gamma (IFN-gamma) has been shown to activate eosinophils and induce expression of membrane receptors. We hypothesized that IFN-gamma mobilizes intracellular RANTES in eosinophils in advance of mediator release. Highly purified peripheral blood eosinophils were obtained from asthmatics and stimulated with IFN-gamma at 500 U/ml for time course analysis up to 2h. By specific ELISA, RANTES was detected in supernatants (80+/-15 pg per 2x10(6) cells) following 120min of stimulation. Immunoreactive RANTES in resting cells (5x10(7) eosinophils) was detected in two intracellular compartments in studies of subcellular fractionation by density gradient centrifugation. After 10 min IFN-gamma stimulation, RANTES immunoreactivity was confined to crystalloid granules. RANTES was redistributed from secretory granules to light-membrane fractions after 60 min of IFN-gamma incubation. Our data suggest that rapid mobilization and release of RANTES occurs from stimulated eosinophils. These findings may have important implications for the role of IFN-gamma in activating human eosinophils, particularly in severe chronic asthma or viral exacerbation of asthmatic inflammation, where this cytokine may play a role.

[Effect of levamisole on microbicidal activity and chemotaxis in polymorphonuclear cells]. / Efecto del levamisol en la actividad microbicida y quimiotaxis en células polimorfonucleares.

AIMS: Neonatal septicemia may have a mortality of 20% in newborns small for their-gestational age. This susceptibility for infections has been explained by immunological disorders. The aim of this study to know the effect of levamisole on the functions of chemotaxis and bacterial activity of PMN cells in newborns small for their-gestational age. MATERIAL AND METHODS: Twenty full-term newborns were studied. Ten of them were small for their-gestational age, and the other 10 had body weight within the normal range. Microbicidal activity and chemotaxis were measured, and comparisons between groups were made with non-parametric statistical tests. RESULTS: In the group of newborns small for their-gestational age. Microbicidal activity was similar to control group, but chemotactic activity was reduced (p < 0.05).

Viladot's operation in the treatment of the child's flatfoot.

We present the results of observations on a series of 77 flat feet treated by Viladot's surgical technique and prosthesis between 1981 and 1989. The correction was evaluated by photopodograms and radiographs. In the final evaluation, 45 cases (58.44%) were judged to have excellent results, 24 (29.87%) good results, 2 (2.59%) fair results, and 7 (9.09%) poor results.

Does the eosinophil have a protective role in amebiasis?

While normal human eosinophils are destroyed in vitro by virulent Entamoeba histolytica, notwithstanding the presence of antibodies and complement, activated eosinophils promptly destroy the parasite although dying also at the end of the process. To study the possible in vivo participation of eosinophils in invasive amebiasis, we compared the induction of experimental amebic abscess of the liver (AAL) in gerbils (Meriones unguiculatus) previously made eosinophilic through Toxocara canis antigen injection and in normal control gerbils. After intraportal inoculation of 10(5) ameba trophozoites (6 and 24 hr), the ratio of gerbils with AAL, as well as the number and size of the microabscesses was comparable in eosinophilic and control gerbils. However, at 96 hr the number and size of the microabscesses were significantly smaller (p < 0.05) in eosinophilic gerbils. On the other hand the actuarial AAL survival curve up to 45 days post-amebic inoculation was significantly (p < 0.05) shifted to the right in controls. These results suggest that antigen-induced eosinophilia may exert a protective effect against AAL in gerbils.

Increased frequency of HLA-DR3 and complotype SCO1 in Mexican mestizo children with amoebic abscess of the liver.

The increase of HLA-DR3 and complotype SCO1 previously found in Mexican mestizo adults with E. histolytica amoebic abscess of the liver, was also found in Mexican mestizo children of either sex with the same disease, when compared to the healthy control population (adults and/or children) of the same ethnic and socioeconomic background. This HLA and complotype pattern was not found in Mexican Mestizo patients with amoebic rectocolitis. No linkage disequilibrium was found between these and the other MHC determinants tested in this survey. Thus, HLA-DR3 and SCO1 may constitute primary, independent risk factors, not for any kind of amoebic tissue invasion (i.e. amoebic rectocolitis), but specifically for amoebic liver abscess, irrespective of age or sex. The possibility of linkage disequilibrium with other factors (i.e. the TNF family) within or close to the MHC that were not tested in this study, is discussed. Children with amoebic liver abscess revealed a significant increase in HLA-DR5, and the absence of HLA-DR6 when compared to adults with amoebic liver abscess, suggesting that at least in this ethnic group these class II HLA traits may contribute to some of the peculiarities of pediatric amoebic liver abscess as opposed to the adult version of this disease. HLA-DR3, SCO1, but also HLA-DR5 and HLA-DR6 have all been associated with certain forms of immune-dysfunction, and may thus contribute to some of the clinical and immunological features of this parasitic disease.

Antigen induced eosinophilia protects gerbils (Meriones unguiculatus) against experimental amebic abscess of the liver.

While the normal human eosinophil is destroyed in vitro by virulent Entamoeba histolytica, notwithstanding the presence of antibodies and complement, activated eosinophils promptly destroy the parasite even though succumbing in the process as well. To study the possible in vivo participation of eosinophils in invasive amebiasis, we compared the induction of experimental amebic abscess of the liver (EAAL) in gerbils (Meriones unguiculatus) previously made eosinophilic (532 +/- 80 eosinophils/mm3) through Toxocara canis antigen injection and normal control gerbils (101 +/- 15 eosinophils/mm3). Shortly (6 and 24 h) after intraportal injection of 10(5) virulent E. histolytica, the ratio of gerbils with EAAL, as well as the number and size of the abscesses was comparable in eosinophilic and control gerbils. At 96 h post-inoculation, the ratio of animals with EAAL was still the same in both groups, yet number and size of abscesses were significantly (p < 0.05) smaller in eosinophilic gerbils. The actuarial EAAL survival curve up to 45 days post-amebic inoculation was significantly (p < 0.05) shifted to the right in eosinophilic gerbils. No significant changes in IL-5 levels were recorded throughout these experiments. The results suggest that antigen-induced eosinophilia may exert a protective effect against EAAL in gerbils. It is speculated that a less overwhelming EAAL strategy--more akin to human amebic abscesses--may reveal this protective effect more clearly.

[In vitro transfer of immunity against PPD with dialyzable extract of leukocytes from human colostrum]. / Transferencia in vitro de inmunidad a PPD con extracto dializable de leucocitos de calostro humano.

The aim of this study is to demonstrate the transference of PPD hypersensibility in an in vitro model, with dialysable colostral leukocyte extract (DCLE) of PPD+ and PPD-mothers, through measurements of leukocyte migration inhibition factor activity (LIF) from blood obtained of the umbilical cord of newborns from PPD+ mothers. The results show that DCLE PPD+ incubated with leukocytes of newborns from PPD- mothers had inhibition of leukocyte migration compared with migration of leukocytes incubated with DCLE PPD-. These results suggest that in this in vitro model, DCLE transfers hypersensibility to PPD.

[Economic development and health]. / Desarrollo económico y salud.

The purpose of this work is to examine the relationship between economic development and the population's level of health. Consequently, we will establish a comparison between Mexico and other countries in various stages of development. The interrelationship between economic development and health is a complex problem which is little understood, even though it is acknowledged that at increased levels of economic development the levels of health of a population will also increase. These considerations can be taken into account when we accept the fact that the health conditions in industrialized countries are generally better than the health conditions in non-industrialized countries. In the case of Mexico, the problems relating to social disparities due to the type of economic development are reflected in the country's level of health. Poor health conditions and the deterioration of health services in Mexico, based on international references, are matters that must be resolved by means of a clear political decision accompanied by the appropriate corresponding resources so that the strategies put forth by the National Health Program (1989-1994) can be carried out.

[Natural history and surgical treatment of pulmonary stenosis. Review of 91 cases]. / Historia natural y tratamiento quirúrgico de la estenosis pulmonar. Revisión de 91 casos.

The natural history of valvular pulmonary stenosis is modified by the surgical treatment of these patients. In order to assess these changes we studied the clinical and hemodynamic features of 91 patients that were treated surgically. Hemodynamic studies were performed postoperatively in 62 of them. All patients had preoperatively a right ventricular systolic pressure above 70 mm Hg and were divided into three groups according to the transvalvular gradient. The first group had less than 50 mm Hg, the second group between 50 and 80 mm Hg, and the third group more than 80 mm Hg gradient. Correlation of these values with asymptomatic (49 patients) and with symptomatic (42 patients) cases showed significant values (p less than 0.001). Postoperatively only 2 patients remained with mild degree effort dyspnea. There was also a significant correlation (p less than 0.005) between the systolic gradient and the cardiothoracic ratio in the studied groups. The T wave in lead VF showed a predictive value in relation to the systolic gradient (p less than 0.001). Reduction of the systolic gradient after surgery showed very significant values when compared to the preoperative gradient (p less than 0.001). The overall mortality was 5.5%.