Fibrous tumors in children - a morphologic and interphase cytogenetic analysis of problematic cases.

We describe and discuss the findings by fluorescent in situ hybridization (FISH) for detection of non-random chromosomal gains, in a group of unusual fibrous lesions in children. Nuclear disaggregation was used to prepare slides from eight cases which were hybridized using alpha-satellite enumeration probes for chromosomes 8, 11 and 17. Trisomy 8 and 11 were detected in a high percentage of nuclei in cases of congenital/infantile fibrosarcomas (ranging from 45 to 80%), and in a low grade fibrosarcoma in an older child (23%). Only gains of chromosome 17 were detected in a case of infantile fibromatosis (22%). In this study we have found that given the unconventional histopathologic features, the detection of more than one non-random chromosomal gains by FISH, may aid in further defining fibrous tumors in children, and may be useful as an ancillary diagnostic test in the future.

Aldosterone regulates Na,K-ATPase and increases lung edema clearance in rats.

Aldosterone increases the Na,K-ATPase function in renal cells involved in active Na(+) transport. Because the alveolar type 2 (AT2) cells participate in active Na(+) transport, we studied whether aldosterone regulates the Na,K-ATPase in rat AT2 cells and whether aldosterone delivered by aerosols to spontaneously breathing rats affects edema clearance in a model of isolated-perfused lungs. The AT2 cells treated with aldosterone had increased Na,K-ATPase beta1-subunit mRNA and protein, which was associated with a 4-fold increase in the Na,K-ATPase hydrolytic activity and the ouabain-sensitive (86)Rb(+) uptake. In physiologic experiments, 24 h after aldosterone was delivered by aerosols to the rat air spaces, the active Na(+) transport and lung edema clearance increased by approximately 53% as compared with control rats and rats in which saline aerosols were delivered. The data suggest that increased active Na(+) transport and lung edema clearance induced by aldosterone is probably due to Na,K-ATPase regulation in alveolar epithelial cells. Conceivably, aldosterone may be used as a strategy to increase lung edema clearance.

Dexamethasone upregulates the Na-K-ATPase in rat alveolar epithelial cells.

Previous studies in kidney, heart, and liver cells have demonstrated that dexamethasone regulates the expression of Na-K-ATPase. In the lungs, Na-K-ATPase has been reported in alveolar epithelial type II (ATII) cells and is thought to participate in active Na+ transport and lung edema clearance. The aim of this study was to determine whether Na-K-ATPase would be regulated by dexamethasone in cultured rat ATII cells. Regulation of the Na-K-ATPase by dexamethasone could lead to a greater understanding of its role in active Na+ transport and lung edema clearance. Rat ATII cells were isolated, plated for 24 h, and exposed to 10(-7) and 10(-8) M dexamethasone. These cells were harvested at 0, 3, 6, 12, and 24 h after dexamethasone exposure for determination of steady-state Na-K-ATPase mRNA transcript levels, protein expression, and function. The steady-state Na-K-ATPase beta1-mRNA transcript levels increased in ATII cells 6, 12, and 24 h after dexamethasone exposure (P < 0.05). However, the steady-state alpha1-mRNA transcript levels were unchanged. The protein expression for the alpha1- and beta1-subunits increased in ATII cells exposed to dexamethasone compared with controls in association with a temporal increase in Na-K-ATPase function after dexamethasone exposure. These results suggest that dexamethasone regulates Na-K-ATPase in ATII cells possibly by transcriptional, translational, and posttranslational mechanisms.

Differential expression of p53, c-kit, and CD34 in prepubertal and postpubertal testicular germ cell tumors.

BACKGROUND: The origin of testicular germ cell tumors (TGCTs) in children is poorly understood. There are clear differences between tumors in young children and those in adolescents and adults, which may suggest that they follow different pathways of tumorigenesis. METHODS: Tissue sections from 25 TGCTs (15 from patients age 4 years or younger and 10 from adolescents or adults) were stained immunohistochemically with anti-p53 (DO-1), CD34, and c-kit proto-oncogene protein product. RESULTS: CD34 expression was noted only in 5 prepubertal tumors. Expression of c-kit was observed in 9 of the 15 prepubertal tumors versus 2 of the 10 postpubertal cases. The intensity of expression was equal to that of the adjacent normal tubules in the prepubertal tumors, whereas the intensity was less in the postpubertal tumors. Expression of p53 was strong in 8 of the 10 tumors in adolescents or adults, with a 40-70% positivity, whereas only 6 of 15 prepubertal tumors expressed p53, with < 10% positivity. CONCLUSIONS: CD34 expression in tumors in the group of young children suggests a possible link to teratomas and further provides insight into the fundamental differences between this group and the adolescent/adult group. The expression of c-kit and p53 provides further evidence that c-kit/SCF signaling and p53 play potentially different roles in the initiation and progression of these tumors. Future studies will be required to clarify this issue.

Ependymomas in children express the multidrug resistance gene: immunohistochemical and molecular biologic study.

In view of the poor response of ependymomas to chemotherapy, it may be hypothesized that these tumors have intrinsic drug resistance to some chemotherapeutic agents. The expression of drug resistance may be specific to a single agent or may involve multiple drugs. Among several mechanisms of drug resistance, P-glycoprotein (Pgp) has been the subject of considerable attention in clinical practice. In order to assess the possible participation of Pgp in the chemotherapeutic resistance of ependymomas, 42 biopsy specimens from 35 patients with ependymoma seen at our institutions were studied by immunohistochemistry with two monoclonal antibodies: C219 (Signet) and UIC-2 (Dr. Roninson's gift). In addition, four cases were studied by polymerase chain reaction after reverse transcription to detect transcripts of Pgp. Our results showed that in 35 samples there was a positive reaction for Pgp with both antibodies; two biopsy samples were positive only with C219 and three others with UIC-2; the remaining two samples were negative with both antibodies. Of the four cases studied by RT-PCR, three showed MDR1 transcripts. These results support our hypothesis of Pgp-mediated intrinsic multidrug resistance in these tumors.

Cellular adhesion molecules in rat adjuvant arthritis.

OBJECTIVE: To examine adhesion molecule expression during the progression of inflammation in a rheumatoid arthritis model of adjuvant-induced arthritis (AIA) in rats. METHODS: Immunohistochemical analysis was used to determine the distribution of the following adhesion molecules: lymphocyte function-associated antigen 1 (LFA-1; CD11a/CD18), Mac-1 and p150/95 (CD11bc/CD18), intercellular adhesion molecule 1 (ICAM-1), and CD44 in tissue sections from the ankle joints of rats with AIA. Control animals and those with AIA were killed at intervals over a 54-day period after injection with mineral oil and Mycobacterium butyricum, respectively. RESULTS: CD44 and LFA-1 were expressed on lymphocytes, macrophages, and synovial (ST) lining cells. CD44 expression on macrophages was found to be increased compared with control animals by day 18, and was significantly increased by day 41. CD44 expression on lymphocytes significantly increased earlier, on days 11-18. Increased LFA-1 expression on macrophages occurred late, on day 41. LFA-1 expression on lymphocytes was significantly increased on days 25, 47, and 54. ST lining cells exhibited two distinct periods of increased expression, one early, on days 11-25 and one later, on days 41-54. CD11b/c was expressed on macrophages and ST lining cells, showing a significant increase on AIA rat ST lining cells compared with control animals on day 4. No differences in ICAM-1 expression on endothelial cells between rats with AIA and controls were found on any of the days examined. CONCLUSION: CD44 expression is up-regulated on macrophages and lymphocytes during the early development of AIA, while LFA-1 expression is up-regulated later in the development of AIA. The up-regulation of CD44 and LFA-1 at different times in the development of AIA suggests an important role for these adhesion molecules in establishing and sustaining an inflammatory response in the AIA joint.

Increased expression of intercellular adhesion molecule 1, CD11/CD18 cell surface adhesion glycoproteins and alpha 4 beta 1 integrin in a rat model of chronic interstitial lung fibrosis.

The expression of the intercellular adhesion molecule 1 (ICAM-1), and the integrins CD49, CD11b/c, and CD11a (LFA-1 alpha chain) was analyzed in an experimental model of pulmonary fibrosis. Adult rats were exposed to 75% oxygen during 10 weeks, and to 2.0 mg/kg of paraquat twice weekly. Rats were sacrificed at 2 days, and at 2 and 10 weeks after the first injection of paraquat. Lungs were fixed in 4% paraformaldehyde and used for histology and immunohistochemistry. At 2 days the lungs showed a diffuse inflammation composed of a mixed polymorphonuclear and mononuclear cell infiltrate. Afterwards, the inflammatory process was predominantly mononuclear, and an increasing fibroblast proliferation was observed. Early inflammatory events (48 h) correlated with a moderate increased expression of ICAM-1, LFA, and CD11b/c in epithelial cells as well as a pronounced expression of ICAM-1 and CD11b/c in macrophages. At 2 and 10 weeks, there was a progressive increased expression of CD11b/c and ICAM-1 by macrophages, as well as of LFA in epithelial cells, and of ICAM-1 and CD49 by epithelial and interstitial cells. Lymphocytes showed a slight increased expression of LFA at 2 weeks, and of CD49 at 2 and 10 weeks. These results suggest that macrophages expressing ICAM-1, CD11b/c, and CD49 are involved in the earlier and late phases of the disease whereas fibroblast and epithelial cells expressing ICAM-1 and CD49 might play a role in the cell interactions involved in the fibrotic phase.

Expression of basic fibroblast growth factor and angiogenin in arthritis.

Angiogenesis is an integral component of the vasculoproliferative phase of rheumatoid arthritis (RA). We examined the distribution of two angiogenic factors, basic fibroblast growth factor (bFGF) and angiogenin (ANG) in arthritic diseases. We used an enzyme-linked immunosorbent assay and immunohistochemical analysis to determine the levels of bFGF and ANG in synovial fluid (SF) and their distribution in synovial tissue (ST), respectively. Most SF contained little or no detectable bFGF ( < 5 pg/ml). ANG in RA SF was 248.7 +/- 17.4 ng/ml, which did not differ significantly from levels found in osteoarthritis (OA; 305.9 +/- 23.1 ng/ml). Synovial lining cells, macrophages, endothelial cells, and vascular smooth muscle cells were immunopositive for bFGF and ANG; however, their expression was not up-regulated in RA ST compared to ST from OA and normal subjects. Though bFGF and ANG are present in the joints of patients with arthritic diseases, they are not up-regulated in RA. These results suggest that not all angiogenic mediators are up-regulated in RA compared to normal subjects and subjects with other arthritic diseases. It may be that some of these mediators, like ANG, play a role in the physiology of normal synovium.

Multidrug resistance gene expression in childhood medulloblastoma: correlation with clinical outcome and DNA ploidy in 29 patients.

Twenty-nine children treated for medulloblastoma between 1987 and 1991 were reviewed. Thirteen patients with high-risk medulloblastoma characterized by incomplete resection, diploid tumor or subarachnoid dissemination received chemotherapy following radiation therapy. Three received postoperative chemotherapy. Eight patients who had been treated with postoperative radiation therapy also received chemotherapy for recurrent tumors. After a minimum 3-year follow-up period, 16 were alive but 13 had died from recurrent tumors. In order to evaluate the possible participation of P-glycoprotein (Pgp)-mediated multidrug resistance (MDR) in medulloblastoma therapy and its correlation with prognosis, archival specimens were examined by immuno-histochemistry utilizing 3 monoclonal antibodies against Pgp and 6 cases by reverse-transcriptase polymerase chain reaction (RT-PCR) using MDR1-specific primers. Sixteen patients (55%) had MDR expression detected either by 1 of the 3 antibodies or by RT-PCR. DNA ploidy study was also performed on 18 specimens. We correlated patients' outcome with variable factors (extent of surgical resection, chemotherapy, DNA ploidy) and MDR expression. Patients who were treated with radiation therapy and adjuvant chemotherapy had a significantly better (p = 0.036) survival than those with radiation therapy alone, despite the fact that the former group of patients was considered to be high-risk. The extent of surgical resection and DNA ploidy did not correlate with prognosis. However, a statistically significant association was found between MDR expression and outcome (p = 0.007). Among the patients who received chemotherapy, positive MDR expression significantly correlated with poor outcome (p = 0.036). Our results showed that Pgp-mediated intrinsic MDR in medulloblastomas seems to correlate with an adverse outcome. This information may be used in designing new therapeutic protocols for medulloblastoma.

Collagenase and collagenase inhibitors in bronchoalveolar lavage fluids.

Collagenase, collagenolytic activity and tissue inhibitor of metalloproteinases were evaluated in bronchoalveolar lavage from 25 patients with hypersensitivity pneumonitis and four control subjects. Patients were followed between two and three years, after which they were classified as "healed," "improved," or "worsened." In control samples, immunoreactive collagenase was not detected. The enzyme was present in four of seven patients who healed, six of ten patients who improved, and four of eight patients who worsened. There was no relationship between the presence or absence of BAL collagenase or its concentration and the evolution of the disease. Latent collagenolytic activity was detected only in 5 of the 14 patients who displayed immunoreactive collagenase. Regarding collagenase inhibitor, TIMP was present in BAL fluid from all patients and normal subjects. Although the highest values were found in two cases who healed or improved, there was not a statistically significant difference among the three groups of patients, neither between patients nor control subjects. These findings suggest that at least in HP, the presence of collagenase, collagenolytic activity, or TIMP in BAL fluid is not associated with the prognosis of the disease.

Clubbing in hypersensitivity pneumonitis. Its prevalence and possible prognostic role.

To know the prevalence and prognostic significance of finger clubbing in hypersensitivity pneumonitis induced by avian antigen, this physical sign was evaluated in 82 patients who were followed up from 1 to 5 years (mean, 2.6 years). According to clinical, roentgenographic, and functional criteria, the patients were classified in one of three stages at admission as well as at least 1 year later. Digital clubbing was retrospectively recorded as present or absent by physical examination. Our results showed that 44 patients (51%) included in this study presented clubbing at the time of diagnosis. Sixteen of these patients presented with worsening of their lung disease, whereas only 5 of the 38 patients without clubbing incurred a worsening of their condition. This finding suggests that digital clubbing is frequent in pigeon breeder's disease and may help to predict clinical deterioration.

Effect of lung T lymphocytes on fibroblasts in idiopathic pulmonary fibrosis and extrinsic allergic alveolitis.

Increased fibroblast replication and interstitial collagen accumulation occur commonly in the interstitial lung disease that progress to fibrosis. The processes controlling lung fibrogenesis are not completely understood, however. This study was designed to analyse the influence of T lymphocytes from lung tissue obtained at open lung biopsy from four patients with idiopathic pulmonary fibrosis and four patients with extrinsic allergic alveolitis on fibroblast proliferation and collagen synthesis in vitro. Lung T cell supernatants from patients with both diseases induced a moderate but significant inhibition of human lung fibroblast cell line growth. In contrast, there was a clear difference in the effect of T cells from the two groups of patients in relation to collagen production. Lung T lymphocytes from all four patients with idiopathic pulmonary fibrosis produced a substantial increase in collagen synthesis (from 371% to 514% of control values), whereas T cells from three of the four patients with extrinsic allergic alveolitis induced a significant decrease in collagen production (to 35%, 36%, and 43% of control values); in the fourth case there was an increase in collagen synthesis but this was lower than that seen with T cells from any of the patients with idiopathic pulmonary fibrosis. Peripheral T cells from six patients and control subjects caused a small increase in fibroblast proliferation and no change in collagen synthesis. The findings suggest that at least two types of interaction occur between lung T cells and fibroblasts in these disorders. A variable degree of inhibition of cell proliferation is observed in response to lung T cell supernatants from patients with both idiopathic pulmonary fibrosis and extrinsic allergic alveolitis; a substantial increase in collagen synthesis is triggered by lymphokines from patients with idiopathic pulmonary fibrosis.

Immunoregulatory abnormalities in patients with pigeon breeder's disease.

T-cell suppression induced by concanavalin-A (Con-A) and the prostaglandin suppressor system (PSS) were studied in 14 patients with pigeon breeder's disease (PBD), 12 and 10 asymptomatic breeders, and 8 controls. Our results showed that PBD patients display a significant decrease in T-cell-induced suppression (29.6 +/- 15.3% vs. 61.2 +/- 9.3% in controls p less than 0.05); whereas asymptomatic breeders respond heterogeneously: 5 showed decreased suppression and 7 were within the normal range obtained in controls. In contrast, the patients presented a higher PSS index compared with the other 2 groups, suggesting an inverse relationship between the 2 systems. These findings indicate that there are relevant differences between PBD patients, asymptomatic breeders, and normal subjects in some immune interactions, which may at least partially explain the characteristic cellular and humoral hyperreactivity observed in patients with this disease.

[Effect of pregnancy and the puerperium on the development of extrinsic allergic alveolitis]. / Influencia del embarazo y puerperio en el desarrollo de la alveolitis alérgica extrínseca.

Extrinsic Allergic Alveolitis (EAA) is a disease characterized by an exaggerated immune response to inhaled organic dusts that can result in alveolar, and occasionally bronchiolar, damage. This damage can disappear completely with the elimination of the antigen and/or treatment with anti-inflammatory agents. Nonetheless, an important group of these patients do not improve; the inflammatory reaction persists. With time, this results in the deposit of abnormal connective, tissue with respect to its quantity and quality, in the pulmonary interstitium destroying the normal parenchymal architecture and making gas exchange impossible. The most common cause of this disease is inhalation of pigeon antigen with a 5:1 predominance in women. Pregnancy has been considered as a case of allotransplantation that produces in the mother a relative hyporeactive condition. In modifying the immune response to the fetus, it is also modified to other foreign antigens. In this work, we studied 80 cases of EAA, 67 females and 13 males. Of the 67 women, 11 (17.1%) presented with the disease after delivery. Symptoms began from 5 days to 7 months after delivery. Eight of the 11 women had been in contact with the antigen before the beginning of the pregnancy without showing symptoms until after delivery. In 4 cases, there had been contact with the antigen in other pregnancies, without symptoms. EAA is a disease involving the immune system in which changes can be seen in its development during pregnancy, as is also seen in other immune diseases, such as Lupus Erythematosus, Rheumatoid Arthritis and other connective tissue diseases. During pregnancy, the mother becomes tolerant. There is a decrease in non-self recognition.(ABSTRACT TRUNCATED AT 250 WORDS)

Paragonimus mexicanus: an antigenic analysis.

El extracto antigenico obtenido de adultos de la especie Paragonimus mexicanus se analiso por inmunodifusion doble y por inmunoelectroforesis empleando el suero hi perinmune de un conejo y los sueros de gatos infectados experimentalmente. Por inmunoelectroforesis se formaron de una banda de identid entre el suero de conejo y los de los gatos. El extracto antigenico contiene trece proteinas, segun fue revelado en un gel de poliacrilamida con dodecil sulfato de sodio