Determinação simultânea de VMA, 5HIAA e HVA urinários sem extração / Simultaneous determination of urinary VMA, 5HIAA and HVA without extraction

Este trabalho mostra as vantagens de se determinar simultaneamente os metabólitos VMA, 5HIAA, HVA urinários. A quantificação destes metabólitos são muito importantes nos diagnósticos e tratamento de tumores que se originam na crista neural (VMA, HVA), e diagnóstico da síndrome e tumor carcinóide (5HIAA). A maioria das técnicas utilizadas, limitam-se a medir somente um ou dois setes metabólitos por análise, sendo que a ESA atualmente desenvolveu um procedimento direto, onde se obtém uma quantificação simultânea dos três metabólitos (VMA, 5HIAA, HVA) urinários usando o Sistema Coulochem Electrode Array – CEAS

Soluble interleukin-2 receptors in sera of children with primary malignant neoplasms.

PURPOSE: T lymphocyte activation regulates the autocrine type secretion of interleukin-2, a T cell growth factor and the de novo expression of its cell-surface receptor (IL-2R). Afterwards, a special, truncated form of the IL-2R is secreted into the serum as a soluble molecule. Soluble interleukin-2 receptor (sIL-2R) in serum is non-specific marker of common activation of the cellular immune regulation. This study reports the results of an early diagnostic observation of sIL-2R level in the sera of 18 primary solid pediatric malignancies. PATIENTS AND METHODS: 18 children between the ages of 1 and 12 represented the Study Group of solid tumor bearing patients. 12 healthy children between the ages of 1 and 12 represented the Control Group. Concentrations of sIL-2R were detected in the sera of the children employing a double-antibody sandwich enzyme-linked immunosorbent assay (T Cell Sciences). RESULTS: Control Group concentrations of sIL-2R were in the range of 101.9 to 255.3 IU/mL with a mean value of 178.6 IU/mL. Serum levels of sIL-2R were markedly elevated in the sera of children with solid tumors (Study Group) to levels between 223.8 IU/mL and 927 IU/mL with a mean value of 575.4 IU/mL. CONCLUSIONS: 1) The shedding of cell-surface receptors of immunological effector cells is a common physiological mechanism; 2) Presence of sIL-2R represents a sign of an overall activation of the effector elements of the host's cellular immune system; 3) An increase in the levels of sIL-2R represents an active phase of progression; conversely, decreasing levels indicate tumor regression; 4) Serum levels of sIL-2Rs may represent a useful laboratory parameter in choosing an efficient anti-cancer therapy. Thus, the level of sIL-2R in childhood solid tumor patients has a prognostic significance.

Morphometric studies of cystic and tubulointerstitial kidney diseases with hepatic fibrosis in children.

Portal tract fibrosis with biliary ductular enlargement or proliferation occurs in a number of genetic diseases that have cystic or tubulointerstitial renal lesions. These include some with renal cystic disease such as autosomal recessive diseases (e.g., infantile polycystic disease, juvenile polycystic disease, and Meckel's syndrome), autosomal dominant diseases (e.g., adult polycystic disease) and, rarely, tuberose sclerosis and dominant glomerulocystic disease. Portal tract fibrosis with biliary enlargement and proliferation occurs also with tubulointerstitial kidney diseases. These probably include at least three disorders in the category nephronophthisis-congenital hepatic fibrosis (one autosomal recessive disease and two either autosomal or X-linked recessive diseases) plus Jeune's syndrome (the tubulointerstitial diseases Fanconi's familial nephronophthisis and anti-tubular membrane antibody disease do not regularly cause hepatic fibrosis). Morphometric data on ratios of bile ductules to connective tissue in hepatic portal tracts show high values for infantile polycystic disease (mean, 0.616) compared to lower values for juvenile polycystic disease (mean, 0.286). That the cystic renal lesions of the first two diseases differ in type and time course is known. Similar data on ratios of glomeruli plus tubules to connective tissue in renal cortices and of tubules to connective tissue in outer medullary zones of kidneys, respectively, are as follows: for Fanconi's nephronophthisis, 0.445 and 0.197; for anti-tubular basement membrane antibody disease, 0.585 and 0.164; and for the three types of nephronophthisis-congenital hepatic fibrosis studied, 0.668 and 0.446, 1.39 and 0.921, and 1.18 and 0.12. These data support clinical impressions that the category nephrophthisis-congenital hepatic fibrosis includes more than one disease entity.

Simultaneous liquid-chromatographic determination of prednisone and prednisolone in plasma.

This high-performance liquid-chromatographic (HPLC) method for simultaneous determination of prednisone and its metabolite, prednisolone, in plasma is a modification of the method of Frey et al. (Clin Chem 1979;25:1944-7). Heparinized plasma (1.0 mL) with 0.1 mL of internal standard solution (11-deoxy-17-hydroxycorticosterone, 2 mg/L) is extracted with 7.0 mL of dichloromethane, then washed sequentially with 0.1 mol/L HCl, 0.1 mol/L NaOH, and deionized water, 2.0 mL each. The extract is evaporated and the residue reconstituted with 75 microL of mobile phase, methanol/H2O (40/60 by vol). Thirty microliters of this is injected onto a reversed-phase C6 column, which is eluted at 1.4 mL/min. Analytical recoveries of prednisone and prednisolone were 94-98% and 102-106%, respectively. Day-to-day precision (CV) was 3.8% for prednisone, 6.1% for prednisolone. We encountered no interference from the 21 other steroids and 25 drugs tested. This method is simple, accurate, and precise.

Comparison of cyclosporine determinations in whole blood by three different methods. HPLC, 125I RIA and 3H RIA.

The authors have analyzed and compared the cyclosporine concentrations in whole blood specimens from pediatric renal transplant patients using three different methods: high-performance liquid chromatography (HPLC) (5u C18 reverse-phase column), 3H radioimmunoassay (RIA) (Sandoz, E. Hanover, NJ), and 125I RIA (substituted 3H-tracer in Sandoz Kit with 125I tracer made by Immuno Nuclear Corporation, Stillwater, MN). Results obtained by the 125I RIA correlated well with results obtained by the 3H RIA. Both RIA methods had similar correlation with the HPLC method. The 125I RIA method showed higher sensitivity and greater precision than the 3H RIA method. The authors conclude that the 125I RIA method can be used for cyclosporine determination in whole blood specimens. The use of the 125I RIA provides a simple and rapid method with higher counting efficiency and less background quenching than the 3H RIA method, which requires cumbersome liquid scintillation counting procedures.

Increased cardiac muscle fiber size and reduced cell number in Down syndrome: heart muscle cell number in Down syndrome.

The size of cardiac muscle fibers was determined for 15 patients with Down syndrome who did not have congenital heart disease and for 15 age- and sex-matched control patients by counting the number of cardiac muscle cells in specified areas of microscopic sections (method of Black-Schaffer and Turner). Mean ratio of muscle cells per unit area for the Down patients versus controls was 84.9%, with mean cross-sectional area of Down fibers 117.8% of control value and calculated mean volume of Down cells 127.7% of control value. Mean weights of Down hearts related to normal for age, versus control hearts, was 79.8% of control values. Calculated ratio of total cardiac muscle cell number in Down hearts was 62.5% of the value for controls. This value corresponds closely to the 62.4%, which can be calculated from Naeye's data on cardiac muscle fiber size in Down syndrome based on a point-count method, and is also close to published values for liver cell size and number and skeletal muscle nuclear number in Down syndrome. This study demonstrates that the Black-Schaffer and Turner method gives results equivalent to those of the point-count method for studies of cardiac muscle fiber size, and further supports the suggestion that chromosome 21 controls the size and number of at least certain cell types.

Microdissection study of the incidence of branched eccrine sweat glands and the number of eccrine glands per unit area of infants' and childrens' skin.

That branched eccrine sweat glands occur has rarely been mentioned in the literature, and their incidence has never been determined. All eccrine glands in 0.5-cm2 pieces of anterior trunk skin of 59 children were microdissected, and the total number of glands and the number of branched eccrine glands determined. One-hundred fourteen of 17,539 (0.65%) were branched, 90% in the middermis and 10% at the epidermis. The apparently normal anatomic property that almost 1% of eccrine sweat glands are branched has not hitherto been appreciated. One doubly branched gland was found. Patients with leukemia (11 in the study) possibly had more branched glands than nonleukemic patients of the same body size. The best statistical relation of the number of sweat glands per unit area of skin (GUA) to surface area (SA) or age in children was the natural logarithm of GUA versus the reciprocal of surface area: LnGUA = (0.2205 X 1/SA) + 5.42. This result is consistent with the classical proposition that there is no important degree of formation of new eccrine glands, nor of loss, after birth, the density of glands per unit area of skin reducing as SA rises with growth during childhood.

The renal lesion of congenital hepatic fibrosis: pathologic and morphometric analysis, with comparison to the renal lesion of infantile polycystic disease.

The renal lesion of congenital hepatic fibrosis (CHF = Blyth and Ockenden's juvenile polycystic disease of liver and kidneys) was analyzed from 6 specimens from patients aged 3 3/12 to 19 3/12 years and compared with that of 5 patients with infantile polycystic disease (IPCD) aged 6 months to 14 4/12 years. Pathologic, microdissection, injection, and morphometric studies show that the predominantly medullary cystic lesion of CHF shows different distribution in medullary, cortico-medullary, and cortical zones of kidney from the lesion of IPCD, and shows a different time course, from early life to renal insufficiency, from that of IPCD. The renal cysts in CHF affect deep or central collecting tubules, in contrast to the involvement of more peripheral orders of collecting tubules in IPCD. Papillary pore counts, performed for 1 patient, gave significantly low values, in contrast to normal values reported for IPCD. The findings support the previously published conclusion, based on differences in the hepatic lesions of the two conditions, that CHF and IPCD are difference diseases, rather than different permissible manifestations of a single disease.

Plasma fibronectin levels after splenectomy and splenic autoimplantation in rats with and without dietary ascorbic acid supplementation.

The critical role of the spleen in protecting subjects from systemic bacterial infection is well known. Plasma fibronectin (PF), cold-insoluble globulin, and opsonic alpha2 surface binding glycoprotein, has regulatory influence on reticuloendothelial system clearance activity and it is important for maximal phagocytosis. Ascorbic acid (AA) also appears to play an important role in phagocytic cell function. The purposes of this study were to determine the effects of splenectomy and splenectomy with autoimplantation, with and without dietary AA supplementation, on PF levels. Six-week-old male Sprague-Dawley rats were divided into four experimental groups of 20 animals each: Nonoperated controls, sham-operated controls, totally splenectomized animals, and splenectomized animals with intraperitoneal autoimplantation. Each group was further randomly divided into two subgroups of 10 animals, those receiving dietary AA supplementation and those not receiving AA. PF levels were measured with a colorimetric assay immediately prior to and at 4 and 8 weeks after operation. Plasma AA determinations documented the effectiveness of dietary AA supplementation. PF levels in nonoperated and sham-operated controls increased significantly during the 8 weeks of experimentation. In contrast, PF levels decreased significantly following total splenectomy from 328 +/- 46 mcg/mL (mean +/- S.D.) to 285 +/- 46 at 4 weeks and rose to 303 +/- 77 at 8 weeks postoperatively in non-AA supplemented animals. Splenic autoimplantation eliminated this decrease in PF levels at 4 weeks. AA supplementation also protected against the decrease in PF levels in the splenectomized group. In the intragroup comparisons, AA supplementation did not produce a significant elevation of PF.(ABSTRACT TRUNCATED AT 250 WORDS)

Eosinophilia in the hospitalized neonate.

The incidence of and clinical associations with eosinophilia in a cross-section of hospitalized newborns had not been studied previously. The medical records of 200 such infants less than 29 days of age were reviewed. Total eosinophil counts were calculated and associations with a variety of clinical factors were examined. Eosinophilia (adjusted for day of age) occurred at least once in 22% of the infants studied. No significant skew could be detected in the age of onset of eosinophilia. Statistical analysis revealed eosinophilia to be significantly associated (p less than .05) with length of hospitalization, number of days of antibiotic therapy and use of parenteral alimentation. A trend toward eosinophilia was noted in lower birth weight infants (0.05 less than p less than 0.10). Eosinophilia was not associated with sex, age, Apgar score at birth, Caesarian section delivery, transfusions, phototherapy, specific diagnoses or type of oral feeding. Multiple regression analysis showed a highly positive correlation (r = .81) of associated factors with hospitalized days as the dependent variable. Eosinophilia appears to be a non-specific finding in sick neonates and is related to the severity of illness and the number of diagnostic or therapeutic procedures performed.

Capnocytophaga sepsis: a newly recognised clinical entity in granulocytopenic patients.

10 episodes of Capnocytophaga sepsis occurred in 6 patients with granulocytopenia, malignancy, and oral mucosal ulcerations. Capnyocytophaga is a newly described genus of gliding gram-negative bacilli which is physiologically and genetically identical to Bacteroides ochraceus and biogroup DF-1 of the United States Center for Disease Control and is commonly responsible for periodontal infections.

Microchemical analysis for 13 constituents of plasma from healthy children.

Normal values for 13 chemical constituents of plasma were estimated from results for 837 presumably healthy children. Ninety microliters of specimen was analyzed for lactate dehydrogenase, aspartate aminotransferase, alkaline phosphatase, inorganic phosphorus, total calcium, total cholesterol, total proteins, albumin, uric acid, urea nitrogen, alanine aminotransferase, total bilirubin, and glucose. We used two Abbott ABA-100 Bichromatic Analyzers interfaced directly to the ABA Data Management System. For each test age- and sex-related variations were assessed and normal values were estimated for six different age groups.

Hyperlipidemia in pediatric hemodialysis and renal transplant patients. Associated with coronary artery disease.

Fasting serum cholesterol and serum triglyceride levels were determined in 15 maintenance hemodialysis (MH) and 35 renal transplant (RT) patients. Fourteen of 15 MH patients (93%) had elevated triglyceride levels (greater than 140 mg/100 ml) compared to 11 of 35 RT recipients (31%) (P less than .001). Two of 15 MH patients (13%) had elevated cholesterol levels (greater than 230 mg/100 ml), compared to 18 of 35 RT recipients (51%) (P = .03). In MH patients, a positive correlation was noted between serum triglyceride levels and carbohydrate intake (P = .03). Autopsy material from 12 children who underwent MH or RT was compared to material from 16 age-matched controls; an increased collagenous content of intima, a possible early indicator of coronary artery disease, was noted more frequently (P less than .006) in index patients compared to controls. Our data demonstrate that hyperlipidemia is a frequent finding in pediatric patients treated with MH and RT, and may be associated with premature coronary artery disease.

Cerebrospinal fluid endotoxin concentrations in gram-negative bacterial meningitis.

The limulus lysate assay was utilized to detect and quantitate endotoxin in cerebrospinal fluid from 232 patients with suspected meningitis. The assay was positive in initial specimens of CSF from all 86 patients with gram-negative bacterial meningitis and was uniformly negative in the remaining 146 patients with a variety of infectious and noninfectious processes. Endotoxin concentrations in initial specimens of CSF from patients with gram-negative meningitis ranged from 4 to 2,000 ng/ml. No correlation between initial CSF levels of endotoxin and initial clinical or laboratory variables of infection was noted. With antibiotic therapy, CSF concentrations of endotoxin fall rapidly to undetectable levels after five days.