Sentinel lymph node in endometrial cancer.

The aim of this study was to evaluate the possibility of identifying the sentinel lymph node and involvement of neoplastic cells in patients with endometrial carcinoma limited to the uterus, and also its correlation with the conditions of other pelvic and para-aortic lymph nodes. Forty patients with endometrial carcinoma, clinical staging I and II, were submitted to complete surgical staging through laparotomy, as recommended by FIGO in 1988. The sentinel node was investigated using patent blue dye in the myometrial subserosa. The sentinel node was excised and submitted to frozen section examination of specimen, stained with hematoxylin and eosin (H&E). Afterward, selective bilateral para-aortic and pelvic lymphadenectomy, total hysterectomy with bilateral salpingo-oophorectomy were performed. The lymph nodes excised were examined by means of paraffin-embedded slices stained with H&E and of imunohistochemistry with antikeratin antibody AE1/AE3. The sentinel lymph node was identified in 77.5% of patients (31/40), and 16.1% (5/31) presented neoplastic involvement in the node. In 25 cases of negative sentinel node, 96% (24/25) had no neoplastic involvement, and 4% (1/25) had other lymph node affected (false negative). In nine cases with no sentinel node identified, 55.5% (5/9) had lymph node involvement. The results of this study allow us to conclude that it is possible to identify the sentinel node using the methods described, and the pathologic examination significantly represents the same conditions of other pelvic and para-aortic lymph nodes.

Identification of the sentinel lymph node stained with blue dye in breast cancer patients.

PURPOSE: This study aimed to evaluate the reproducibility of sentinel lymphadenectomy in breast cancer patients (T1N0M0 and T2N0M0) and its possibility of predicting the total axillary behavior. METHODS: A total of 25 patients were evaluated, all presenting palpable mammary nodes between 1.5 and 5 cm (T1 and T2), with clinically negative axillary lymph nodes (N0). After an incisional biopsy of the tumor and histopathological confirmation of invasive breast carcinoma, a study of the sentinel lymph node took place with a peritumoral injection of 4 ml of blue dye at 2.5%. After waiting for 15 to 20 minutes, a search for the blue stained lymphatic vase in the axillary fat was carried out, which would lead to the sentinel lymph node, stained or not. At that point, a mastectomy (20 patients) or a quadrantectomy (5 patients) was performed, both with axillary lymphadenectomy at grades 1, 2 and 3. The sentinel lymph nodes and the material from the axillary dissection were sent separately for an anatomicopathological test in paraffin. RESULTS: The lymph nodes were identified in 19 patients, which represented a 76% detection rate. There was a concordance between the sentinel lymphadenectomy and the standard axillary dissection in 68.4% of the patients. The false-positive and the false-negative rates observed were 10% and 55.5%, respectively. A higher detection rate was found in tumors larger than 2 cm and situated in external quadrants. CONCLUSIONS: Sentinel lymphadenectomy identified the sentinel lymph node in the majority of the patients in this study, although the high rate of false-negatives observed prevented an accurate staging.

Deficiency of vitamins E and A in cystic fibrosis is independent of pancreatic function and current enzyme and vitamin supplementation.

UNLABELLED: The aim of this study was to evaluate to what extent serum vitamins A and E cystic fibrosis are affected by the underlying disease, pancreatic sufficiency or insufficiency, meconium ileus, nutritional status, age and treatment (enzyme and vitamin supplementation). Serum vitamin A and E levels were determined by high performance liquid chromatography in 210 cystic fibrosis patients, subdivided according to clinical condition into four subgroups (unsupplemented pancreatic insufficiency, supplemented meconium ileus, pancreatic sufficiency, supplemented pancreatic insufficiency) and compared with 42 control subjects. Vitamin A and E levels were generally lower in cystic fibrosis patients than in controls (P < 0.002 and P < 0.001 respectively). Subjects with pancreatic insufficiency regularly receiving enzyme and vitamin supplementation had significantly lower vitamin A (P < 0.05) and vitamin E (P < 0.01) levels than controls. In subjects with pancreatic sufficiency only vitamin A was significantly lower than in controls (P < 0.01). Vitamin levels were not age-dependent in cystic fibrosis, and no significant correlation with standardized body weight (Z-score) was observed. CONCLUSION: Cystic fibrosis patients show a clear tendency to vitamin A and E deficiency, irrespective of pancreatic function, body weight and standardized supplementation with pancreatic extract and liposoluble vitamins. Since the clinical significance of this deficiency is still not clear, longitudinal studies of cystic fibrosis patients with and without adequate vitamin supplementation are required.

Assessment of neutrophil aggregation by Coulter STKR and STKS haematological analysers.

We have studied an alternative method to aggregometry for the assessment of human polymorphonuclear (PMN) leucocyte aggregation. This simple, rapid and reliable procedure counts unaggregated cells on both Coulter STKS and STKR haematological analysers by the impedance principle. Aggregation of PMN was induced by 15 min incubation with fresh autologous serum (FAS) after a 10 min phorbol myristate acetate (PMA) activation of neutrophils in small aliquots (0.25 ml) of suspension containing about 4.0 x 10(9) PMN/1. Differences (x 100) between count of resting and PMA+FAS treated neutrophils/count of resting PMN reflect percent aggregation. By this procedure, PMN aggregation did not occur in autologous plasma from EDTA anticoagulated whole blood; it was partially inhibited by hydrocortisone, whereas inactivated or Zymosan activated sera gave values similar to those from FAS induced aggregation. PMA aggregation was dependent on Ca2+ + Mg2+ concentration. Intra-assay analytical variability did not exceed 4% on either instrument. Reference values (n = 20) of percent PMN aggregation were 50.7 +/- 4.7 on STKS and 47.1 +/- 4.8 on STKR. Most probably, the interindividual variance was due to the physiological variability of Mg2+ and/or Ca2+ concentrations in FAS. Thus, this procedure reflects the true PMN aggregability status in a given subject, and in a given electrolyte environment.

High light scatter by neutrophils in the Bayer-Technicon H*2 analyzer: a screening test of morphologically defective responsiveness to in vitro chemotactic stimulation.

The Bayer-Technicon H*2 haematological analyser provides differential white blood cell count, including the assay of polymorphonuclear leukocytes by light scattering and the absorbance increase following the cytochemical reaction for myeloperoxidase. The mean value of polymorphonuclear leukocytes scatter, which reflects polymorphonuclear leukocytes volume, is printed in a separate report "for laboratory use only" as a ybar value in arbitrary units. In certain patients neutrophils displayed an unreported correlation between polymorphonuclear leukocytes high ybar basal values (> or = 37.00 arbitrary units) (determined on the H*2) and a defective response in vitro to the chemoattractant, formyl-methionyl-leucyl-phenylalanine (determined by microscopic evaluation of polymorphonuclear leukocytes shape change (polarization)). The patients showing no polymorphonuclear leukocyte response or a defective one to formyl-methionyl-leucyl-phenylalanine were all affected by "Systemic Inflammatory Response Syndrome (SIRS)". Therefore the predictive value of the positive test for SIRS is 100%. On the other hand 8.8% of SIRS patients had polymorphonuclear leukocytes < 37.00 arbitrary units of ybar basal value and a "normal" response to formyl-methionyl-leucyl-phenylalanine; the predictive value of the negative test being 90%. Since we demonstrated in vitro a dose-dependent deactivation of endotoxin or lipopolysaccharide-pretreated polymorphonuclear leukocytes, the "normal" response to formyl-methionyl-leucyl-phenylalanine of the "false negative" cases may occur because the endotoxaemia in these patients is too low to prevent it.(ABSTRACT TRUNCATED AT 250 WORDS)

A simple assessment of human neutrophil adhesiveness.

The adhesiveness of human polymorphonuclear leukocytes was assessed in serum-coated polystyrene spectrophotometric cuvettes. Capped cuvettes, containing no more than 2 x 10(6) resting or concanavalin A-treated (100 micrograms/ml) polymorphonuclear leukocytes, were laid horizontally and subjected to three 90 degrees rotations on their major axis at fixed times. After incubation at room temperature, non-adherent cells remaining in suspension were counted on the Coulter counter STKS hematological analyzer. After a 16-min incubation (4 min each side of the cuvette) the adhesion of concanavalin A-activated neutrophils ranged from 98% to 100% and the adhesion of resting neutrophils from 30% to 35% (mean 32.4 +/- 2.2%, n = 10). An 8-min incubation (2 min each side) led to approximately 50% adhesion of concanavalin A-activated neutrophils (mean 49.9 +/- 2.2%, range 46%-54%, n = 16), whereas the adhesion of resting cells was about 21% (mean 21.4 +/- 1.6%, range 19%-24%, n = 16). The variation in percentage adhesion in repeated assays did not exceed 4% using concanavalin A-activated cells and 7.5% with resting neutrophils. The procedure is very rapid, easy to perform and precise, and no special apparatus or glassware is necessary. The method also allows microscopic evaluation of shape changes of adherent neutrophils through the clear sides of the cuvettes.

Multicenter evaluation of Reflotron direct dry-chemistry assay of high-density lipoprotein cholesterol in venous and fingerstick specimens.

The Reflotron HDL Cholesterol test (Boehringer Mannheim GmbH) directly separates and analyzes high-density lipoprotein (HDL) cholesterol in plasma collected with EDTA in an integrated dry-reagent system suitable for alternative site testing of lipoproteins. We describe a multicenter evaluation of this test by two US and six European laboratories experienced in lipid analysis. Each laboratory compared the Reflotron with the same conventional wet-chemistry method, Boehringer phosphotungstate-Mg2+ precipitation with enzymatic cholesterol assay. Imprecision was within accepted guidelines, with CVs of < or = 8% for fresh and frozen plasmas (median CV 1.7-3.9%) and for lyophilized sera (median CV 3.8-4.7%), similar to those of the conventional method. Results of linear-regression analysis were as follows: Reflotron HDL Cholesterol = 1.03 conventional - 3.9 mg/L, r = 0.987. The Reflotron results were somewhat low in the two US laboratories, demonstrating the need for general standardization of methods for measuring HDL cholesterol. Results from capillary fingerstick plasma agreed well with those from venous-derived plasma; capillary = 1.04 venous + 4.5 mg/L, r = 0.967. The system is relatively insensitive to interference from hemoglobin (< or = 0.75 g/L), ascorbic acid (< or = 0.3 g/L), bilirubin (< or = 50 mg/L), cholesterol (< or = 3.5 g/L), and triglycerides (< or = 4 g/L). The relative ease of operation and the rapid availability of results (within 90 s for plasma collected in EDTA) make the method appropriate for use by well-trained, but not necessarily technical, operators in the physician's office or other alternative sites.

Volumetric changes in phorbol myristate acetate activated neutrophils: a rapid and simple assay using Coulter counter STKR and STKS hematological analyzers.

BACKGROUND: The aim of this study was to investigate some physical changes (volume, conductivity and scatter) in human polymorphonuclear leukocytes (PMN) activated with phorbol myristate acetate (PMA). METHODS: Volume changes in PMA-activated neutrophils were assayed by both STKR and STKS, two Coulter hematological analyzers. Scatter changes in PMN activated by PMA in suspensions containing nitro-blue tetrazolium (NBT) were investigated on STKS scattergrams. RESULTS: PMA activation induced PMN volumetric increases that could be assayed with STKR and displayed with STKS. The activation of PMN in suspension containing NBT induced cellular scatter changes on STKS scattergrams. The differences in scatter between resting and PMA-activated neutrophils may thus provide a semiquantitative assay of NBT reduction and superoxide production. Volume changes in PMA-activated neutrophils were due to cellular swelling through water uptake, induced by Na+/H+ antiport activation and Na+ influx. Since volume changes in PMA-activated neutrophils might occur without O2- production and vice versa, in the cases in which PMA activation of protein kinase C cannot be demonstrated by O2- production, the effect of PMA on protein kinase C- mediated Na+/H+ antiport activation, and in turn on PMA volume changes, reflects protein kinase C activation. CONCLUSIONS: A screening of neutrophils unresponsive to volumetric changes from PMA activation may be easily performed using both Coulter STKR and STKS analyzers, whereas expected or defective PMA-induced production of O2- may be semi-quantitatively evaluated by STKS.

Serum low density lipoprotein separation: a simple procedure.

Reference methods for serum low density lipoprotein (LDL) separation are time-consuming and the salts used in density-gradient ultracentrifugation may cause chemical and/or immunological changes in the lipoprotein structure. A method has been developed to provide native LDL suitable for chemical and immunochemical studies. The three step procedure involves firstly the separation of very low density lipoproteins by non-density adjusted ultracentrifugation, secondly the separation of low from high density lipoproteins by LDL precipitation with PEG-6000, and finally the re-dissolution of the pellet in NaCl 150 mmol/l. The effectiveness of LDL separation, as well as the preservation of the electrophoretic mobility of the LDL molecules, was verified by electrophoresis in agarose gel, and the maintenance of the immunochemical reactivity of apolipoprotein B was verified by an immunochemical assay.

Volume, conductivity, and scatter changes of activated polymorphonuclear leukocytes: an estimation by Coulter Counter STKS analyzer.

Suspensions of phorbol myristate acetate-activated polymorphonuclear leukocytes were analyzed with a Coulter Counter STKS hematological analyzer. Phorbol myristate acetate activation induced an increase in polymorphonuclear leukocyte volume and conductivity, while scatter was unchanged. Phorbol myristate acetate-activated neutrophils in a suspension containing nitroblue tetrazolium showed increased scatter. The rise in scatter was phorbol myristate acetate dose dependent, completely inhibited by diphenylene iodonium and partially by dimethyl sulfoxide, two inhibitors of NADPH oxidase. Zymosan-activated polymorphonuclear leukocytes were notably larger with a characteristic position on discriminant function 1 display (volume versus scatter) of the analyzer. Volume and conductivity changes were seemingly inexplicable features of phorbol myristate acetate activation. The rise in scatter was produced by cytoplasmic precipitation of reduced nitroblue tetrazolium and thus by O2-generation in phorbol myristate acetate-activated neutrophils. Zymosan phagocytosis was responsible for the notable rise in polymorphonuclear leukocyte volume. The analysis of activated polymorphonuclear leukocytes by Coulter Counter STKS may provide useful information on their activation and a pragmatic approach for studying function.

Advantages of a new anticoagulant in routine hematology on the Coulter Counter S-Plus STKR analyzer.

The authors report the advantages of a new anticoagulant-antiaggregant mixture that avoids the deleterious effects of ethylenediaminotetraacetic acid (EDTA) on mean platelet volume and also prevents EDTA-induced platelet clumping. It is suitable for routine cell counting and sizing with the Coulter Counter S-Plus STKR. The values of the common hematologic parameters agree well with those from EDTA-treated samples and are stable for at least eight hours after sampling.

EDTA-induced platelet aggregation can be avoided by a new anticoagulant also suitable for automated complete blood count.

In vitro EDTA-induced platelet aggregation is a fairly rare event but can have serious clinical consequences producing pseudothrombocytopenia and pseudoleukocytosis. Sixteen specimens with previously recognized EDTA-induced platelet aggregation were collected in a new anticoagulant-antiaggregant mixture containing trisodium citrate 17 mmol/l, pyridoxal 5'-phosphate 11.3 mmol/l and Tris 24.76 mmol/l (CPT mixture) and analyzed at various times after venepuncture with four hematological instruments: Coulter Counter S-Plus STKR, Technicon H6000, Technicon H1 and Ortho ELT-8. In CPT-anticoagulated specimens the signals and instrumental flags of platelet clumping were absent, and the platelet number correlated very well with a microscopic count from a finger stick drawn into Unopette. The complete blood count was very similar in "normal" hematological specimens either collected in K3. EDTA or in CPT, although Technicon H1 and Ortho3 ELT-8 required a suitable calibration for MCV and hematocrit in the latter mixture. Mean platelet volume was stable for up to 24 h only in CPT-collected specimens, if it was measured on a Coulter Counter S-Plus STKR. In routine hematological practice CPT can be an alternative anticoagulant to K3. EDTA, most suitable for automated complete blood count and useful in avoiding EDTA-induced platelet clumping.

[Obstetric factors associated with low birth weight]. / Fatores obstétricos associados ao baixo peso ao nascer.

An analysis was made of 18,804 of 19,446 consecutive births of the number analysed 15.93% presented low birth-rate. Significant statistical association was found in relation to maternal age, pre-natal care, previous pregnancies, smoking and gestational age at birth. Measures with a view to the attenuation of the problem are proposed, among them being: educational programs for teenagers on human reproduction, programs designed to create awareness of the harm done by smoking, amplification of antenatal assistance, medical programs for the limitation of premature labor, all of these and others, in association with programs of socio-economic support.

[Myasthenia gravis and pregnancy: review of the literature and report of 6 cases occurring in 5 patients]. / "Miastenia gravis" e gestação: revisão da literatura e relato de seis casos ocorridos em cinco pacientes.

The authors report six cases of myasthenia gravis during pregnancy. Three patients experienced increasing severity of their disease and one death occurred in the puerperium. The infants were born with no evidence of neonatal myasthenia. The patients should be closely monitored during labor and puerperium. This disease is characterized by unpredictable exacerbations, and the association of myasthenia gravis and pregnancy increases the risk for the patient.

[Cerclage of the cervix uteri via a transabdominal route: report of 2 cases]. / Circlagem do colo uterino por via transabdominal: relato de dois casos.

The authors present two cases of transabdominal cervicouterine cerclage using Benson and Durfee's technique. In both cases the surgery was successful and the patients gave birth at 37 and 40 weeks, respectively, to healthy newborns. The technique is recommended by the authors since technical details are observed.