The advancement of a new human factors report--'The Unique Report'--facilitating flight crew auditing of performance/operations as part of an airline's safety management system.

This paper presents the findings of research relating to the specification of a new human factors report, conducted as part of the work requirements for the Human Integration into the Lifecycle of Aviation Systems project, sponsored by the European Commission. Specifically, it describes the proposed concept for a unique report, which will form the basis for all operational and safety reports completed by flight crew. This includes all mandatory and optional reports. Critically, this form is central to the advancement of improved processes and technology tools, supporting airline performance management, safety management, organisational learning and knowledge integration/information-sharing activities. Specifically, this paper describes the background to the development of this reporting form, the logic and contents of this form and how reporting data will be made use of by airline personnel. This includes a description of the proposed intelligent planning process and the associated intelligent flight plan concept, which makes use of airline operational and safety analyses information. Primarily, this new reporting form has been developed in collaboration with a major Spanish airline. In addition, it has involved research with five other airlines. Overall, this has involved extensive field research, collaborative prototyping and evaluation of new reports/flight plan concepts and a number of evaluation activities. Participants have included both operational and management personnel, across different airline flight operations processes. Statement of Relevance: This paper presents the development of a reporting concept outlined through field research and collaborative prototyping within an airline. The resulting reporting function, embedded in the journey log compiled at the end of each flight, aims at enabling employees to audit the operations of the company they work for.

Integrated stereological and biochemical studies on hepatocytic membranes. III. Relative surface of endoplasmic reticulum membranes in microsomal fractions estimated on freeze-fracture preparations.

New methods are required for identifying membranes in subcellular fractions with respect to their origin, if such preparations are to be evaluated morphometrically. One method is freeze-fracturing which reveals intramembrane particles whose size, pattern, and numerical density differ for various membrane types. The question is examined whether the differences in numerical particle density per square micrometer of membrane (alpha) can be used to differentiate membrane vesicles found in microsomal fractions from liver cells with respect to their origin in the hepatocytes. It is found that the range of alpha for the protoplasmic face (PF) of endoplasmic reticulum (ER) membrane (1,900 less than alpha less than 3,250) is intermediate between those for plasma and mitochondrial membranes. Since PF(ER) should appear in the outer leaflet of microsomal vesicles, alpha was estimated on concave profiles of freeze-fracture preparations; the numerical frequency distribution of vesicles with respect to alpha was trimodal, with a major peak around 2,900/micrometer2 and 66% of the vesicles in the range determined for PF(ER). Using a new stereological method, it was calculated that 63% of the membrane surface in these microsomal fractions was of ER origin by this criterion. On the same preparations, an attempt was made to label the ER-derived membranes cytochemically for glucose-6-phosphatase. A line intersection count revealed 62% of the membrane surface to be of ER origin on the basis of marker enzyme labeling. These findings indicate a smaller part of ER membranes in microsomal fractions than would be predicted from biochemical data (77%). The possible reasons for such discrepancies are discussed; shifts in particle densities due to the preparation procedure could lead to an underestimate by freeze-fracturing, whereas the prediction from biochemical data could be overestimates if marker enzymes were not homogeneously distributed.

Integrated stereological and biochemical studies on hepatocytic membranes. I.V. Heterogeneous distribution of marker enzymes on endoplasmic reticulum membranes in fractions.

The purpose of the study was to consider quantitatively the relationships between the surface area of the endoplasmic reticulum (ER) and constituent marker enzyme activities, as they occur in fractions collected from rat liver homogenates. The ER surface area was estimated in five membrane-containing fractions by use of a combined cytochemical-stereological technique (5), while, at the same time, ER marker enzymes were assayed biochemically. Fraction/homogenate recoveries for the ER enzymes averaged 100%, total membrane surface area 98%, and ER surface area 96%. Relative specific activities, which compare the relative amounts of ER marker enzyme activities to the relative ER surface area in the membrane-containing fractions, indicate variable distributions for glucose-6-phosphatase and NADPH cytochrome c reductase, but not for esterase.

Intergrated stereological and biochemical studies of hepatocytic membranes. I. Membrane recoveries in subcellular fractions.

Previous attempts to relate the structure and function of hepatocytic membranes have compared biochemical data of fractions to morphological data derived from either intact tissue or fractions. The effects of the original homogenization aside, biochemical recoveries comparing membrane marker enzymes of the homogenate to subsequent fractions suggest a general conservation of activity. A sterological study was undertaken to estimate membrane surface areas in the intact tissue, homogenate, and fractions of the same livers and then to test the comparability of these data with membrane marker enzymes by calculating both morphological and biochemical recoveries. The sterological data were corrected for errors due to section thickness and compression. The average total membrane sufrace area per 1 g of liver was 9.3 m2 in the intact tissue (T), 7.8 m2 in the homogenate (H), and 7.4 m2 in the fractions (F); recoveries for the membrane surface areas thus averaged 96% for the (F/H) and 81% for the (F/T) comparisons. In homogenate and fractions, the differentiability of membranes by morphological criteria was limited to rough- and smooth- surfaced membranes, as well as outer and inner mitochondrial membranes. The recoveries of rough-surfaced membranes were 101% for F/H and 92% for F/T; those of smooth-surface membranes were 89% for F/H and 107% for F/T. For mitochondrial membranes, a recovery of 100% for F/H was obtained, whereas it amounted to only 54% for F/T. With respect to F/H, the membrane recoveries compare well with the marker enzyme recoveries obtained biochemically. The extension of recovery calculations to the intact tissue (F/T) revealed satisfactory conservation of the procedures of homogenization and fractionation; it indicates, however, that a shift of a substantial part of mitochondrial membranes to the pool of unidentifiable smooth membranes may occur on homogenization.

Detection of the common acute lymphoblastic leukemia antigen in the serum of leukemia patients.

The common acute lymphoblastic leukemia antigen (CALLA) has been detected in biological fluids using a radioimmunoassay based on the inhibition of binding of 125I-labeled monoclonal anti-CALLA antibody to glutaraldehyde-fixed NALM-1 cells. With this assay, we showed first that CALLA was released in culture fluids from NALM-1 and Daudi cell lines but was absent from culture fluids from CALLA negative cell lines. Then, we found that the sera of 34 out of 42 patients (81%) with untreated common acute lymphoblastic leukemia (c-ALL) contained higher CALLA levels than any of the 42 serum samples from healthy controls. The specificity of these results was further demonstrated by testing in parallel the sera from 48 patients with CALLA negative leukemias, including 26 acute myeloid leukemia (AML), 12 T-cell acute lymphoblastic leukemia (T-ALL), and 10 acute undifferentiated leukemia (AUL). All of these sera gave negative results, except for one patient with AUL, who had a significantly elevated circulating CALLA level, and one patient with AML, who had a borderline CALLA level, 3 SD over the mean of the normal sera. Preliminary results suggest that circulating CALLA is associated with membrane fragments or vesicles, since the total CALLA antigenic activity was recovered in the pellet of the serum samples centrifuged at 100,000 g. In addition, the CALLA-positive pellets contained an enzyme considered as a membrane marker, 5'-nucleotidase. Evaluation of the clinical importance of repeated serum CALLA determinations for the monitoring of c-ALL patients deserves further investigation.

Partial characterization of proteoglycans isolated from neoplastic and nonneoplastic human breast tissues.

The biochemical composition of proteoglycans was investigated in human breast tissues of different age either with invasive mammary carcinoma or with benign lesions of the breast. Proteoglycans were extracted from tissues under dissociative conditions (4 M guanidine-HCl), isolated by CsCl gradient ultracentrifugation, and purified by gel exclusion and ion exchange chromatography. Glycosaminoglycan side chain compositions of proteoglycans were evaluated by enzymatic analysis (chondroitinases ABC and AC) and nitrous acid degradation. Biochemical data indicated that proteoglycans of high density and molecular size were increased (per wet weight of tissue) in neoplastic compared to nonneoplastic tissues. Overall proteoglycan content was increased almost 2-fold in tumors. Furthermore, enzymatic data revealed a change in the proportions of glycosaminoglycan chains in neoplastic and nonneoplastic tissues. In particular, an increase in chondroitin sulfate (63% versus 35%, respectively) together with a decrease of dermatan sulfate (12% versus 45%, respectively) characterized tumors in comparison to mammary tissues with benign lesions, while the relative content of heparan sulfate side chains remained similar in both tissues. However, morphometric analyses revealed that heparan sulfate content per epithelial cell volume was in fact decreased in neoplastic tissue. These differences in proteoglycans indicate that there are significant changes in the extracellular matrix and surface properties of cells in breast cancer tissue.

Fractal organization of feline oocyte cytoplasm.

The present study aimed at verifying whether immature cat oocytes with morphologic irregular cytoplasm display self-similar features which can be analytically described by fractal analysis. Original images of oocytes collected by ovariectomy were acquired at a final magnification of 400x with a CCD video camera connected to an optic microscope. After greyscale thresholding segmentation of cytoplasm, image profiles were submitted to fractal analysis using FANAL++, a program which provided an analytical standard procedure for determining the fractal dimension (FD). The presentation of the oocyte influenced the magnitude of the fractal dimension with the highest FD of 1.91 measured on grey-dark cytoplasm characterized by a highly connected network of lipid droplets and intracellular membranes. Fractal analysis provides an effective quantitative descriptor of the real cytoplasm morphology, which can influence the acquirement of in vitro developmental competence, without introducing any bias or shape approximation and thus contributes to an objective and reliable classification of feline oocytes.

Enzymatic imbalance in peripheral blood mononuclear cells isolated from individuals with anti-HIV antibodies.

Plasma membrane-bound 5'-nucleotidase (5'-NT), gamma-glutamyltransferase (gamma-GT) and soluble deoxynucleotidyltransferase (TdT) were studied in peripheral blood cells (PBMN) of 35 individuals, 26 male and 9 female, with circulating anti-HIV antibodies. Twenty-six were drug abusers, 2 were drug abusers and homosexuals and 4 were homosexuals. Three did not fall into any risk group. The surface immunologic phenotype of cells stained with the fluorescent monoclonal antibodies Leu 5, Leu 3, Leu 2, Leu 12, Leu M3, Leu M1, anti-CALLA and anti-HLA-DR was delineated by flow cytometry. While the gamma-GT activity did not change, the lymphocyte 5'-NT activity was significantly less than normal in anti-HIV positive individuals and in anti-HIV negative drug abusers. TdT activity was detectable in 14 anti-HIV positive patients (40%), who did not have clinical AIDS. Of 8 patients with AIDS, 3 had a low level of TdT activity but 5 had cells completely devoid of TdT and 5'-NT activity. 5'-nucleotidase activity and the frequency of Leu 2 suppressor antigen bearing cells were the only independent variables that correlated with AIDS incidence.

Ultrastructural aspects of endometrial surface in Kartagener's syndrome.

The endometrium of an infertile patient with Kartagener's syndrome showed initial secretory phase characteristics at SEM, whereas TEM analysis demonstrated several alterations in the central and peripheral microtubular distribution in 87% of the cilia examined. Such aspects seemed appropriate for a normal implantation, but the ciliary immotility or dyskinesia could cause an altered flow of the endometrial secretions and compromise the upstream movement of the spermatozoa.

Secretory changes in preovulatory endometrium during controlled ovarian hyperstimulation with buserelin acetate and human gonadotropins.

Twenty-one patients with unexplained infertility underwent controlled ovarian hyperstimulation with buserelin acetate and human menopausal and chorionic gonadotropins, and follicular growth was monitored by ultrasonography and daily 17 beta-estradiol (E2) assays. Endometrial biopsy was performed when E2 levels were greater than or equal to 250 pg/mL per follicle and the follicular diameter was greater than or equal to 17 mm. As controls, we studied 20 preovulatory endometrial biopsies from patients with a male infertility factor. The biopsy material was examined at light, scanning, and transmission electron microscopes, and a morphometric analysis was performed. Preovulatory endometrial mucosa during controlled ovarian hyperstimulation showed accentuated proliferative aspects in both the glandular and stromal components and more frequent early secretory phenomena compared with the controls. The preovulatory progesterone (P) levels observed in our study seem to exclude the possibility that such early secretory aspects of the hyperstimulated endometrium are because of higher P concentrations in patients compared with controls.

Relationship of steroid hormone receptors to the cloning of fresh breast cancer tissues.

Thirty-seven samples of human primary breast cancer were processed for direct cloning in methylcellulose (MC) cultures. Out of the 37 specimens plated, 19 (51%) tumors grew with a plating efficiency (PE) of 0.012%. Both growing and nongrowing tumors belonged mostly to the ductal histological type. Neither the use of autologous serum (AS) nor of fetal calf serum (FCS) affected the PE. Moreover, a negative correlation was found between the level of estrogens and especially of progestin receptors and the ability of tumors to grow in MC culture. These findings underline the difficulty of cloning fresh specimens of human solid tumors and indicate that malignant cells displaying high concentrations of progestinic receptors may also display a degree of differentiation which leads to a reduced clonogenic ability.

Treatment of fallopian tube carcinoma with cyclophosphamide, adriamycin, and cisplatin.

Primary carcinoma of the fallopian tube is uncommon; optimal primary treatment is still not well defined, and little information is available about the efficacy of cisplatin-based combination chemotherapy. Thirty-eight patients with fallopian tube carcinoma were treated with cyclophosphamide (500 mg/m2), Adriamycin (50 mg/m2), and cisplatin (50 mg/m2) (CAP). Thirty-two patients received the combination chemotherapy as first-line treatment after cytoreductive surgery, whereas six subjects were treated for recurrent disease. The patients received a median of six cycles of therapy (range, four to nine). At the initiation of chemotherapy, 24 patients had measurable lesions. In this group of patients, 15 had a clinical complete response (CR), four had a partial response (PR), three had stable disease (SD), and two had progressive disease (PD) after chemotherapy. The overall clinical response rate (CR + PR) was 80%. Ten of the 14 CR patients who were submitted to second-look operation (SLO) were found free of disease, in pathologic complete response (pCR). Three pCR patients relapsed, and two of them died despite second-line treatment. Nine patients achieving PR, SD, and PD after first-line chemotherapy were further treated (five with chemotherapy, two with radiotherapy, two with progesteron), but none responded to second-line treatment and all died (median survival, 9 months). Fourteen patients without gross residual disease after cytoreductive surgery had no measurable lesions and were not evaluable for response. Seven of them had negative SLO and remain disease free. Three patients (two stage III and one stage II) who refused SLO relapsed 14, 16, and 26 months after completion of chemotherapy. The median survival for the entire group was 38 months, and the 5-year survival rate was 35%. The toxicity of the regimen was moderate. The CAP regimen appears to be active in primary fallopian tube carcinoma and yields response rates comparable to those reported for epithelial ovarian cancer.

Freeze-fracture morphometry on normal and cancer breast tissues.

A stereological procedure to evaluate the number of particles per unit surface area (Np/micron2) and the particle density distribution in both protoplasmic (pf) and external face (ef) of the plasma membrane was applied on freeze-fracture preparations of human breast tissues with invasive carcinoma and with non-neoplastic lesions. A significant higher intramembrane particle density was recorded on the protoplasmic face of plasma membrane of epithelial cells from non-neoplastic breast tissues (2999 +/- 720 Np/micron2) compared to the particle density on the corresponding face of cancer cells (911 +/- 608 Np/micron2). Moreover, in the latter cells the external face had more than 50% of the surface area totally smooth while residual particles were often aggregated. The reduction of integral glycoproteins in the plasma membrane of malignant tissues fits apparently well with the increased activity of the methyltransferase-I, an enzyme methylating membrane phospholipids. These findings suggest a distribution of constituents and a lipid environment peculiar for the plasma membrane of breast cancer cells.

Fractal dimension of pericellular membranes in human lymphocytes and lymphoblastic leukemia cells.

When blood cells are projected into an image plane their contours appear as borderlines of irregular shape with the property of the statistical self-similarity. The dimension D of these cell perimeters may be fractal rather than an integer as in the Euclidean space. The fractal dimension D for different q'order moments was evaluated by means of a probabilistic method after recording EM image profiles of normal peripheral blood mononuclear cells (PBMN) and mature lymphocytes and also of lymphoblasts isolated from humans with acute leukemia or obtained by lectin stimulation in vitro. PBMN cells, CD 4 and CD 8 T-lymphocytes showed a fractal dimension D quite similar among five moments, ranging from 1.23-1.17. Cells of hairy-cell leukemia with highly convoluted morphology displayed D comprised between 1.32-1.36, whereas blasts of T or B acute lymphoblastic leukemia were characterized by a smaller D of 1.11 and 1.13-1.16 respectively. When normal T-lymphocytes were transformed into blasts by lectin PHA stimulation in vitro, the fractal dimension D was significantly reduced and found close to the value recorded on pericellular contour of blasts from acute T-lymphoblastic leukemia.

Effect of autologous serum and steroid hormone receptors on direct cloning of human breast cancer.

Thirty-seven samples of human primary breast cancer were processed for direct cloning in methylcellulose (MC) cultures. Of the 37 specimens plated, 19 tumors (51%) grew with a plating efficiency (PE) of 0.012%. Both growing and nongrowing tumors belonged mostly to the ductal histologic type. The use of autologous serum v fetal calf serum did not affect the PE. Moreover, a negative correlation was found between the level of estrogen receptors and especially of progesterone receptors (PRs) and the ability of tumors to grow in MC culture. These findings underline the difficulty of cloning fresh specimens of human solid tumors and indicate that malignant cells with a high concentration of PRs may also have a degree of differentiation that leads to a reduced clonogenic ability.

Steroid hormones modify nuclear heterochromatin structure and plasma membrane enzyme of MCF-7 cells. A combined fractal, electron microscopical and enzymatic analysis.

Ultrastructural features of the nuclear membrane envelope (ENM) and the nuclear membrane-bound heterochromatin (NMBHC) were investigated in MCF-7 human breast cancer cells by fractal morphometry. The fractal dimension D established by the box counting method proved to be effective for quantifying nuclear changes in MCF-7 cells treated with steroid hormones, namely the estrogen 17 beta-estradiol, which stimulates cell proliferation, and the glucocorticoid dexamethasone. When MCF-7 cells were briefly (5 min) cultured in the presence of 17 beta-estradiol (10(-9) M), the irregularity of the NMBHC outline was increased as documented by the increased fractal dimension D. Changes in the ultrastructural complexity of the nuclear heterochromatin were observed in concomitance with functional changes at the cell periphery, namely the modulation of the estrogen-induced activity of phospholipase C, a cell membrane-associated enzyme involved in the signal transduction pathway via phosphoinositides metabolism. Dexamethasone did not affect the in vitro proliferation, the phospholipase C activity nor the shape of the ENM of MCF-7 cells, but reduced the structural complexity of the nuclear membrane-bound heterochromatin.

Limited invasiveness to assess retroperitoneal spread in stage I-II ovarian carcinoma.

OBJECTIVE: To evaluate the incidence of retroperitoneal metastases, survival rate and site of recurrence in early ovarian tumors undergoing limited retroperitoneal surgery. METHOD: Three hundred seventy-three consecutive patients underwent assessment of the retroperitoneum consisting of intraoperative palpation with or without biopsies. RESULTS: Retroperitoneal metastases were detected in 10 stage-I tumors (3.2%) and in 10 stage-II tumors (16%). The risk was inversely related to tumor differentiation. Palpation revealed metastases in 10 cases. During follow-up, none of the borderline tumors (1.9% of stage-I grade-1 node-negative, 2.7% of grade-2 and 7.0% of grade-3 tumors) recurred in the retroperitoneum. In stage II, two recurrences were observed in grade-2 tumors (11%) and one in grade 3 (4.5%). CONCLUSION: Limited retroperitoneal surgery enables satisfactory outcome in early ovarian cancers. Risk of retroperitoneal recurrence is minimal in grade 1 and non-existent in borderline tumors. Less differentiated tumors have low risk but further investigation of the therapeutic role of lymphadenectomy is justified.

Resveratrol modulates apoptosis and oxidation in human blood mononuclear cells.

BACKGROUND: We examined the effect of resveratrol (RS), a nonflavonoid polyphenolic phytoalexin found in grapes and red wine, and RS coincubated with the oxidant 2-deoxy-D-ribose (dR), on apoptosis and on the oxidative metabolic status of normal human peripheral blood mononuclear cells (PBMNCs) isolated ex vivo from healthy donors. MATERIAL AND METHODS: Apoptosis was measured by changes of membrane permeability to propidium iodide (PI), plasma membrane exposure of phosphatidylserine (PS) and intracellular caspase activity. Oxidative status was assessed by recording the intracellular glutathione concentration (GSH), the activities of the enzymes y-glutamyltransferase (y-GT) and glutathione-S-transferase (GST), and intracellular lipid peroxidation (MDA). RESULTS: Neither apoptotic nor oxidative parameters were affected by culturing PBMNCs in medium containing RS up to 20 micro M for 5 days, while the frequency of cells with intermediate permeability to PI (17% +/- 5) increased at 50 micro M of RS. Thus resveratrol was slightly toxic, but there was little apoptosis in these cells. Peripheral blood mononuclear cells were also grown first in medium plus RS for 24 h and then for 96 h in medium containing RS plus 10 mM of dR, an oxidant sugar that is apoptogenic for human lymphocytes. The apoptotic changes triggered by dR were counteracted by the phytoalexin in a dose-dependent manner, but RS activity was absent at the lowest concentration (5 micro M) and significantly reduced at the highest concentration used (50 micro M). In PBMNCs coincubated with 20 micro M of RS and 10 mM of dR the antioxidant effect of RS manifested with a significant reduction of caspases-3, -8, y-GT, GST activities and MDA content. CONCLUSIONS: Peripheral blood mononuclear cells acquire antioxidant capacity when treated with RS. Grape resveratrol may make a useful dietary supplement for minimizing oxidative injury in immune-perturbed states and human chronic degenerative diseases.

Fractals in pathology: are they really useful?

Through several examples selected from functional and pathological cells and tissues, including surface, cytoplasmic and nuclear membranes one will focus on the application of the fractal approach to measuring irregular and complex structures, on the importance of evaluating real morphology and on its potential role in understanding of tumor biology.