OBJECTIVE: To describe the latest version of SisPorto, a program for automated analysis of cardiotocograms that closely follows the FIGO guidelines, analyses ante- and intrapartum tracings, performs no signal reduction, and has the possibility of simultaneously recording twins. METHODS: A detailed description of the program's processing algorithms and operation is provided, as well as the main results of the studies performed to-date with this system. RESULTS: Considering both current and previous versions of the program, SisPorto has been tested in over 6000 pregnancies. The system's FHR baseline was compared with an average of three experts' estimates, and the difference was under 8 bpm in all cases. A fair to good agreement was found with experts' identification of accelerations, decelerations, contractions, and normal/reduced variability (proportions of agreement 0.64-0.89). In a preliminary validity study (n = 85), a sensitivity of 100% and a specificity of 99% were obtained in prediction of poor neonatal outcome. The system is currently undergoing an international multicentre validation study. CONCLUSIONS: Although still at the research level, a considerable experience has now been gathered with this system. Promising results have been achieved in studies comparing SisPorto with experts' analysis and in those evaluating the validity of the system.

Algorithms↗

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Metastatic calcifications and severe hypercalcemia in a patient with parathyroid carcinoma.

Primary hyperparathyroidism is rarely produced by parathyroid carcinoma. We present the case of a 63-yr-old man who was admitted due to recent onset of constipation, weakness and progressive lethargy. At physical examination, a left cervical mass was palpated. Marked hypercalcemia (serum calcium 25 mg/dl) (6.22 mmol/l) complicated by renal insuficiency (serum creatinine 4.4 mg/dl) (388 micromol/l) was found, but both were unresponsive to conventional therapy and hemofiltration. Autopsy examination showed a carcinoma of the upper left parathyroid gland, multiple foci of metastatic calficications in the vessel walls and parenchyma of both lungs and kidneys, and the myocardium, which contributed to multi-organ failure and death. In addition to describing the clinical presentation, we review the mechanism of metastatic calcifications as well as the role of renal function and hyperphosphatemia, and the basis for therapy of hypercalcemic crisis.

Autopsy↗

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[Detection of Brucella melitensis by BACTEC 9050 system].

OBJECTIVES: To evaluate BACTEC 9050 system capacity for detection of bacteremia due to Brucella spp. MATERIAL AND METHODS: 16 febrile patients were studied during an epidemic infection for Brucella spp. Suspicious of brucellosis was serologically confirmed with Rose Bengal test and agglutination tube test. Only one blood culture was processed per patient. Ten millilitres of blood were inoculated in a Bactec Plus aerobic/F bottle and incubated in BACTEC 9050 automatic system. The bottles were kept in incubation during 21 days, and they were subcultured when the machine detected its growth; if not, a blind subculture was performed after 21 days. RESULTS: 13 of 16 patients showed bacteriemia (81.2%): 11 patients were detected by BACTEC 9050 system and 2 patients by blind subculture after 21 days. A positive result appeared in 149.8 hours (6.2 days) as a mean. Earlier detections were seen in 83 hours (3.4 days) and the latest ones at 245 hours (10.2 days). The aetiology agent of the epidemic infection was Brucella mellitensis biotype 1. We found significative differences comparing the BACTEC 9050 with BACTEC 9120/9240 systems (p<0.03). CONCLUSIONS: Incubation protocols of 5 days are not useful for BACTEC 9050 system in the case brucellosis suspicious. Protocols of 7 would detect only 69.2% of the cases.

Agglutination Tests↗

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Immunogenicity of H-2 positive and H-2 negative clones of a mouse tumour, GR9.

The GR9 tumour was induced with methylcholanthrene in a BALB/c mouse, adapted to tissue culture and cloned without any passage in vivo GR9 clones were typed for H-2 with three monoclonal antibodies that define H-2Kd + Dd, Kd and Dd antigens. A great heterogeneity of H-2d expression was found from clones which were Kd and Dd positive to clones Kd and Dd negative. These results were confirmed for A7 and B9 clones using immunoprecipitation with anti-H-2D.4 and anti-H-2K.31 alloantisera and SDS-PAGE analysis. In addition, the number of chromosomes per cell was heterogeneous amongst the clones, ranging from 38 +/- 2 to pseudotetraploid clones which have 75 +/- 2 chromosomes. GR9 clones were injected into syngeneic BALB/c mice to measure local tumour growth. We found that the growth correlated with the amount of H-2 antigen expressed, i.e. clones with low H-2d expression were highly malignant while clones with normal H-2d expression were highly immunogenic. Finally we found that BALB/c mice immunized against A7 (Kd, Dd-positive) protected against A7, as expected, but surprisingly also against B9 (Kd, Dd-negative).

Animals↗

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Characterization of the syngeneic anti-tumour response against the GR9 tumour with the production of isoantisera and monoclonal antibodies.

Isoantisera and syngeneic monoclonal antibodies have been produced against A7, a clone derived from a BALB/c methylcholanthrene-induced tumour named GR9. The antigens defined by the isoantisera and mAbs are widely distributed in a large number of GR9 clones as well as in other tumour cell lines of different aetiology and origin. Two GR9 clones with marked differences in H-2 expression and syngeneic in vivo growth (A7, which is H-2d-positive and highly immunogenic and B-9, which is H-2d-negative and highly malignant) were selected to further analyse the presence or absence of the antigens recognized by the syngeneic anti-tumour antibodies. Immunoprecipitation data with two different isoantisera revealed a group of proteins with a molecular weight of Mr 65-70,000 present in both clones. Furthermore, immunoprecipitation with the A7-1 mAb again showed a single Mr 65K band present in A7 and B9 clones. We conclude that the marked differences observed in the syngeneic growth of the two clones are not due to differences in tumour associated antigens (TAA), defined by syngeneic antibodies, and emphasize the role that class I H-2 antigens could play in tumour rejection.

Animals↗

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Generation of syngeneic anti-tumour double negative cells (CD4- CD8-), with cytotoxic activity against clones of different MHC class I expression.

In vivo immunization and secondary culture techniques were used to generate cytotoxic T lymphocytes against a tumour cell clone obtained from a methylcholantrene-induced fibrosarcoma. Our CTLs differed from classical CTLs (Ly2+ and MHC restricted) in that our lymphocytes exhibited a non-MHC restricted cytotoxic activity directed to the original tumour and to other MCA-induced tumours, but have failed to demonstrate any killing activity against a wide range of tumour cell lines of diverse origin (including NK sensitive targets). Depletion and phenotypic studies demonstrated that these cells bear the Thyl.2 antigen but are negative for both Ly2 and L3T4 antigens. These CTLs may belong to a double negative subset (CD4-, CD8-) involved in the anti-tumour response.

Animals↗

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Virtual automation.

Total laboratory automation (TLA) can be substituted in mid-size laboratories by a computer sample workflow control (virtual automation). Such a solution has been implemented in our laboratory using PSM, software developed in cooperation with Roche Diagnostics (Barcelona, Spain), to this purpose. This software is connected to the online analyzers and to the laboratory information system and is able to control and direct the samples working as an intermediate station. The only difference with TLA is the replacement of transport belts by personnel of the laboratory. The implementation of this virtual automation system has allowed us the achievement of the main advantages of TLA: workload increase (64%) with reduction in the cost per test (43%), significant reduction in the number of biochemistry primary tubes (from 8 to 2), less aliquoting (from 600 to 100 samples/day), automation of functional testing, drastic reduction of preanalytical errors (from 11.7 to 0.4% of the tubes) and better total response time for both inpatients (from up to 48 hours to up to 4 hours) and outpatients (from up to 10 days to up to 48 hours). As an additional advantage, virtual automation could be implemented without hardware investment and significant headcount reduction (15% in our lab).

Autoanalysis↗

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