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Biomedical subjects

M Cardoso

Publications and source records attributed to M Cardoso.

At least 19 recordsLinked to original sources

Computed tomographic simulation of craniospinal fields in pediatric patients: improved treatment accuracy and patient comfort.

PURPOSE: To reduce the time required for planning and simulating craniospinal fields through the use of a computed tomography (CT) simulator and virtual simulation, and to improve the accuracy of field and shielding placement. METHODS AND MATERIALS: A CT simulation planning technique was developed. Localization of critical anatomic features such as the eyes, cribriform plate region, and caudal extent of the thecal sac are enhanced by this technique. Over a 2-month period, nine consecutive pediatric patients were simulated and planned for craniospinal irradiation. Four patients underwent both conventional simulation and CT simulation. Five were planned using CT simulation only. The accuracy of CT simulation was assessed by comparing digitally reconstructed radiographs (DRRs) to portal films for all patients and to conventional simulation films as well in the first four patients. RESULTS: Time spent by patients in the CT simulation suite was 20 min on average and 40 min maximally for those who were noncompliant. Image acquisition time was <10 min in all cases. In the absence of the patient, virtual simulation of all fields took 20 min. The DRRs were in agreement with portal and/or simulation films to within 5 mm in five of the eight cases. Discrepancies of > or =5 mm in the positioning of the inferior border of the cranial fields in the first three patients were due to a systematic error in CT scan acquisition and marker contouring which was corrected by modifying the technique after the fourth patient. In one patient, the facial shield had to be moved 0.75 cm inferiorly owing to an error in shield construction. CONCLUSIONS: Our analysis showed that CT simulation of craniospinal fields was accurate. It resulted in a significant reduction in the time the patient must be immobilized during the planning process. This technique can improve accuracy in field placement and shielding by using three-dimensional CT-aided localization of critical and target structures. Overall, it has improved staff efficiency and resource utilization.

Adolescent

Correction of maternal serum bile acid profile during ursodeoxycholic acid therapy in cholestasis of pregnancy.

BACKGROUND/AIMS: Intrahepatic cholestasis of pregnancy is characterized by pruritus and increased levels of serum bile acids, and is often associated with premature delivery, fetal distress, and perinatal mortality. The aims of the present study were: (i) to better define the serum bile acid profile in intrahepatic cholestasis of pregnancy and its potential usefulness for differential diagnosis; (ii) to investigate the effect of ursodeoxycholic acid treatment on the bile acid pool; and (iii) to investigate possible adverse effects of therapy. METHODS: Fifteen patients with intrahepatic cholestasis of pregnancy were enrolled in this study. Ursodeoxycholic acid (14 mg/kg body weight per day) was administered for 13 +/- 5 days. Twenty normal pregnant women served as controls. Serum bile acid profile was analyzed by high-performance liquid chromatography. RESULTS: Patients with cholestasis of pregnancy showed significant alterations in the proportion of primary bile acids, with an increase in cholic acid (64.0 +/- 3.0% vs. 32.2 +/- 1.8%, p < 0.01), and a decrease in chenodeoxycholic acid (20.8 +/- 1.4% vs. 31.9 +/- 1.3%, p < 0.01), as compared to controls, resulting in a marked elevation in the cholic/chenodeoxycholic acid ratio (3.4 +/- 0.5 vs. 1.1 +/- 0.1, p < 0.01). The glycine/taurine ratio was reduced in cholestasis of pregnancy (0.8 +/- 0.1 vs. 1.4 +/- 0.1, p < 0.01). During ursodeoxycholic acid administration its proportion in serum increased from 1.4 +/- 0.6% (0.6 +/- 0.2 micromol/l) at baseline to 24.7 +/- 2.3% (5.9 +/- 1.9 micromol/l) with therapy (p < 0.01). This increment was accompanied by a significant decrease in the percentage of cholic acid (28.2 +/- 2.6%, p < 0.01) and an elevation in chenodeoxycholic acid proportion (25.0 +/- 1.9%, N.S.). Although lithocholic acid concentration in serum was maintained with treatment (1.2 +/- 0.2 micromol/l vs. 1.7 +/- 0.5 micromol/l), there was a significant increase in lithocholic acid proportion (p < 0.01) from 3.3 +/- 0.5% at baseline to 7.4 +/- 1.3% during therapy. The glycine/taurine ratio of serum bile acid pool returned to normal after ursodeoxycholic acid administration (1.7 +/- 0.3). CONCLUSIONS: These results establish the importance of ursodeoxycholic acid treatment for the correction of maternal serum bile acid profile in cholestasis of pregnancy, indicating that ursodeoxycholic acid may improve fetal prognosis.

Adult

Staphylococcal strains involved in bovine mastitis are inhibited by Staphylococcus aureus antimicrobial peptides.

The inhibitory activity of five bacteriocin (Bac)-producer strains of Staphylococcus aureus was tested against bacteria pathogenic for cattle. Sixty-five epidemiologically unrelated strains of Staph. aureus involved in bovine mastitis were used as indicators in an agar diffusion test. Bacteriocins produced by four strains could inhibit only a limited number of test organisms. However, all 65 indicator strains proved to be susceptible to the combined action of both bacteriocins encoded by pRJ9, a Bac plasmid found in strain A53. Therefore, the bacteriocins produced by this strain may represent new antimicrobial peptides with potential applications in the prevention and treatment of bovine mastitis.

Animals

Estimated risk of transmission of hepatitis C virus by blood transfusion.

OBJECTIVE: The risk of transmitting hepatitis C (HCV) by transfusion of anti-HCV-negative screened blood was estimated for the blood donor population of Baden-Württemberg (southwestern Germany). METHODS: The data from the blood donors screened for anti-HCV and for HBsAg during 1990-1995 were analyzed. RESULTS: The prevalence of confirmed anti-HCV -positive blood donations decreased continuously during the last 5 years, reaching 121 per 100,000 blood donations. A higher anti-HCV prevalence rate was found in female than in male blood donors (p < 0.05). The estimated risk of transmitting HCV during the window period is 1:200,000 (1:97,000-1:1,400,000) for repeat donors. In 1995, the calculated risk for first-time donors was 1:20,000 (1:15,000-28,000). The incidence for HCV was 1.2 per 100,000 blood donations. CONCLUSION: The risk of transmitting hepatitis C by blood transfusion is low. Additional tests to shorten the window period to detect antibodies to HCV might increase the safety of blood transfusion.

Blood Donors

Autologous platelet transfusion in alloimmunized patients with acute leukemia.

Seventy-eight transfusions of autologous platelets were given to eight alloimmunized patients receiving curative chemotherapy for acute leukemia. Platelets were collected at regeneration of hematopoiesis after a chemotherapy cycle, cryopreserved with 5% dimethylsulfoxide in liquid nitrogen, and retransfused during bone marrow aplasia following the next treatment cycle. The in vitro platelet recovery after freezing, thawing, and washing was 85 +/- 4%. The in vivo corrected count increment 1 h after autologous platelet transfusions was 11 +/- 5 x 10(9)/l. With the exception of moderate urticaria and slight nausea each after one transfusion, no immediate or chronic side effects occurred. The bleeding time was shortened and hemorrhage during bone marrow aplasia was prevented in all alloimmunized patients by autologous platelet transfusions.

Acute Disease

Quality of pooled platelet concentrates prepared from buffy coats and stored in an additive solution after filtration.

Platelet concentrates prepared from buffy coat were pooled and stored for 6 days after removal of leukocytes by filtration. The platelets were stored in plasma or in an additive solution, Plasmalyte-A. In vitro platelet function was better preserved using Plasmalyte-A than plasma with regard to osmotic reversal and aggregation. No significant differences for the release of platelet markers beta-thromboglobulin, platelet factor 4, or lactate dehydrogenase pre- and post-filtration and storage in plasma or Plasmalyte-A was observed. Expression of the surface membrane glycoproteins Ib, Ia/IIa, IIb/IIIa, and IV measured by flow cytometry after binding of monoclonal antibodies did not change during storage. The expression of activation-dependent alpha-granula glycoprotein GMP140, the thrombospondin, and the glycoprotein 53 from the lysosomal granules was not different between platelet pools stored in plasma or in Plasmalyte-A. The in vitro quality of platelets stored as pools is comparable for plasma and the additive solution Plasmalyte-A.

Acetates

In vitro platelet function during storage in three different additive solutions.

Three different synthetic media without glucose were studied for platelet storage. The first medium contained acetate and gluconate. The second contained acetate, gluconate and citrate. Finally the third contained phosphate and mannitol. The purpose of the study was to investigate whether there were differences among the various media in terms of preservation of platelet quality. Pools of platelet concentrates were prepared from buffy coats. In vitro function and metabolic parameters were measured during 5 days of storage in these additive solutions as well as in plasma. Platelet aggregation, hypotonic shock response and release of beta-thromboglobulin, platelet factor 4 and lactate dehydrogenase of the cytosol were equivalent in the media containing acetate compared to plasma storage. In vitro platelet functions and pH in these two media were better preserved compared to the medium with phosphate and mannitol. In addition bacteriological studies using platelets suspended in additive solutions or in plasma were carried out. Carryover of 20% of plasma to the synthetic media necessary for successful platelet storage in these additive solutions allows bacteriological growth. As shown, inoculation of 1 colony/ml Staphylococcus epidermidis leads to 10(6)-10(7) organisms/ml after 5 days of storage.

Blood Donors

[Recombinant antibodies in serologic diagnosis].

In this paper we discuss the value of a new methodology for the production of monoclonal antibodies by recombinant techniques. This approach is especially useful for human monoclonal antibodies due to instability of human antibody-producing cell lines. As an example, we present a recombinant human antibody to the hepatitis C virus (HCV) core protein.

Antibodies, Monoclonal

[Risk of infection transmission by blood components].

OBJECTIVE: Assessment of the present risk of blood-borne infection associated with homologous blood transfusion in Central Europe with particular emphasis on HIV and HCV infections. DATA SOURCES: The relevant literature in the English and German language and the authors' data. STUDY SELECTION: No special study has been carried out for the present paper. RESULTS: The recognition of the risk for transmitting HIV by transfusion has led to a bundle of measures which have improved the safety of blood supply in Germany. A stricter donor selection as well as screening for HIV 1 and HIV 2 have reduced the risk to transmit HIV to the order of 1 per 1 million units transfused. The transmission of hepatitis B is estimated to be in the order of 1:50,000. The anti-HCV testing, introduced in 1990, has markedly reduced the transmission of HCV to less than 1:5,000 per unit as judged from our own data. A further reduction is expected by an improved HCV screening in the near future. Fatal disease from bacterial contamination is rare, with an estimated risk of one in a million units. Syphilis transmitted by transfusion is virtually not occurring anymore. However, increased efforts should continue to enhance the safety of blood, bearing in mind that a zero risk is not achievable for effective therapies. The estimation of risks is a dynamic, time-dependent value which has to be estimated for a geographically defined population for a given time period. CONCLUSIONS: The risk of transfusion-associated infection has been markedly reduced in recent years. Albeit there is a small but definitive risk, which is often overestimated. The risks have to be defined to estimate the risk/benefit ratio of the homologous as well as the autologous transfusion for the individual patient.

Blood Banks

Chloramphenicol resistance plasmids in Staphylococcus aureus isolated from bovine subclinical mastitis.

Chloramphenicol resistance (CmR) could be detected in 11 of 217 Staphylococcus aureus isolates from bovine subclinical mastitis. All isolates were assigned to biotypes A or C. The CmR-determinants were found to be located exclusively on small plasmids of approximately 4.6 kb as revealed by protoplast transformation. The 11 CmR-plasmids could be differentiated on the basis of restriction endonuclease analyses. The restriction maps of these CmR-plasmids identified two separate groups. One group demonstrated homology to the plasmid pC 221, the other to the plasmid pC 223. Both prototype plasmids, pC 221 and pC 223, had been isolated from S. aureus of human origin.

Animals

Characterization of the chloramphenicol acetyltransferase variants encoded by the plasmids pSCS6 and pSCS7 from Staphylococcus aureus.

The two 4.6 kb chloramphenicol resistance (CmR) plasmids pSCS6 and pSCS7, previously identified in Staphylococcus aureus from subclinical bovine mastitis, both encoded an inducible chloramphenicol acetyltransferase (CAT, EC 2.3.1.28). The pSCS6- and pSCS7-encoded CAT variants were purified by ammonium sulphate precipitation, ion-exchange chromatography and fast protein liquid chromatography (FPLC). Both native enzymes showed Mr values of 70,000 on FPLC and were composed of three identical subunits, each of Mr approximately 23,000. The CAT variants from pSCS6 and pSCS7 differed in their net charges and in their isoelectric points. The isoelectric point of the CAT from pSCS6 was pH 5.7 and that of the CAT from pSCS7 pH 5.2. Both CAT variants exhibited highest enzyme activities at pH 8.0. The Km values for chloramphenicol and acetyl-CoA of the CAT from pSCS6 were 2.5 microM and 58.8 microM, respectively, while those of the CAT from pSCS7 were 2.7 microM and 55.5 microM. Both CAT variants were relatively thermostable. The CAT from pSCS6 was less sensitive to mercuric ions than the CAT from pSCS7.

Amino Acid Sequence

Nucleotide sequence and structural relationships of a chloramphenicol acetyltransferase encoded by the plasmid pSCS6 from Staphylococcus aureus.

The 4.6 kb chloramphenicol resistance (Cm) plasmid, pSCS6, isolated from a naturally occurring Staphylococcus aureus biotype C encoded an inducible chloramphenicol acetyltransferase (CAT). The respective cat gene and its regulatory region were cloned. Sequence analyses revealed two open reading frames: one for a 9-amino acid leader peptide and the other for the 215-amino acid CAT monomer. Comparisons of the predicted CAT amino acid sequences revealed a high degree of similarity between CAT from pSCS6 and the CAT variants encoded by Cm plasmids of the pC221 family. These close structural relationships suggested an intraspecific exchange of Cm-determinants between Staph. aureus of human and bovine biotype.

Amino Acid Sequence

Nucleotide sequence and phylogeny of the tet(L) tetracycline resistance determinant encoded by plasmid pSTE1 from Staphylococcus hyicus.

The nucleotide sequence of the tetracycline resistance (tet) gene and its regulatory region, encoded by the plasmid pSTE1 from Staphylococcus hyicus, was determined. The tet gene was inducible by tetracycline and encoded a hydrophobic protein of 458 amino acids. Comparisons between the predicted amino acid sequences of the pSTE1-encoded Tet from S. hyicus and the previously sequenced Tet K variants from Staphylococcus aureus, Tet L variants from Bacillus cereus, Bacillus stearothermophilus, and Bacillus subtilis, Tet M variants from Streptococcus faecalis and Staphylococcus aureus as well as Tet O from Streptococcus mutans were performed. An alignment of Tet amino acid sequences revealed the presence of 30 conserved amino acids among these Tet variants. On the basis of the alignment, a phylogenetic tree was constructed. It demonstrated large evolutionary distances between the Tet M and Tet O variants on one hand and the Tet K and Tet L variants on the other hand. The pSTE1-encoded Tet proved to be closely related to the Tet L proteins originally found on small Bacillus plasmids. The observed extensive similarities in the nucleotide sequences of the tet genes and in the deduced Tet amino acid sequences allowed the assignment of the pSTE1-encoded Tet to the Tet proteins of class L.

Amino Acid Sequence

Cryopreserved autologous platelet transfusions in alloimmunized patients with acute leukemia.

Autologous platelets of 5 alloimmunized patients with acute leukemia in remission were cryopreserved with 5% dimethylsulfoxide in liquid nitrogen and retransfused in the following therapy-induced thrombocytopenic phase. The mean platelet recovery after freezing, thawing and washing was 85 +/- 6%. The mean corrected 1-hour increment in platelet counts was 11 (4-27) x 10(9)/l, i.e. 61% in comparison with fresh platelet transfusions in other patients. In vivo function of frozen platelets was documented by improvement of the posttransfusion bleeding time. Cryopreserved autologous platelets function hemostatically and can be used even for completely refractory patients and thus permit curative antileukemic therapy.

Adult

[The application of the problem-solving method in a situation of surgical intervention; a case report].

This study used the problems solving method for directing nursing assistance to the hospitalized surgical patient. We present the elaborate instrument founded in the Bailey e Claus model which is analyzed and exemplified. The authors consider the available resources concluding that the proposed is possible to be established. They realize how necessary is in nursing all the taking decision process to reach the proposed objectives, toward the resolution of the patient problems.

Brazil

Cloning and sequence analysis of a plasmid-encoded chloramphenicol acetyltransferase gene from Staphylococcus intermedius.

The chloramphenicol acetyltransferase gene (cat) of a 3.9 kb chloramphenicol resistance (CmR) plasmid from Staphylococcus intermedius, designated pSCS1, was cloned into an Escherichia coli plasmid vector. Sequence analysis revealed a high degree of base similarity with the cat gene of the S. aureus CmR plasmid pC221 but there were several differences in the regulatory region. A lesser degree of similarity was observed between the cat gene of the S. intermedius plasmid and the cat gene of the S. aureus plasmid pC194.

Amino Acid Sequence

Molecular cloning, purification, and properties of a plasmid-encoded chloramphenicol acetyltransferase from Staphylococcus haemolyticus.

A small chloramphenicol resistance (Cmr) plasmid of approximately 3.75 kb, designated pSCS5, was isolated from Staphylococcus haemolyticus. This plasmid encoded an inducible chloramphenicol acetyltransferase (CAT; EC 2.3.1.28). The cat gene of pSCS5 was cloned into the Escherichia coli plasmid vector pBluescript SKII+. It differed in its nucleotide sequence and deduced amino acid sequence from the cat genes described previously in staphylococci and other gram-positive bacteria. The CAT enzyme was purified from cell-free lysates by ammonium sulfate precipitation, ion-exchange chromatography, and fast protein liquid chromatography. The native enzyme had an Mr of 70,000 and was composed of three identical subunits, each with an Mr of approximately 23,000. Its isoelectric point was at pH 6.15. CAT from pSCS5 exhibited Km values of 2.81 and 51.8 microM for chloramphenicol and acetyl coenzyme A, respectively. The optimum pH for activity was 7.8. CAT encoded by pSCS5 proved to be relatively heat stable, but sensitive to mercury ions. The observed differences in the nucleotide sequence and the biochemical characteristics of the enzyme allowed the identification of the pSCS5-encoded CAT from S. haemolyticus as a CAT variant different from those described previously in gram-positive bacteria.

Amino Acid Sequence

Nucleotide sequence and phylogeny of a chloramphenicol acetyltransferase encoded by the plasmid pSCS7 from Staphylococcus aureus.

The nucleotide sequence of the chloramphenicol acetyltransferase gene (cat) and its regulatory region, encoded by the plasmid pSCS7 from Staphylococcus aureus, was determined. The structural cat gene encoded a protein of 209 amino acids, which represented one monomer of the enzyme chloramphenicol acetyltransferase (CAT). Comparisons between the amino acid sequences of the pSCS7-encoded CAT from S. aureus and the previously sequenced CAT variants from S. aureus, Staphylococcus intermedius, Staphylococcus haemolyticus, Bacillus pumilis, Clostridium difficile, Clostridium perfringens, Escherichia coli, Shigella flexneri, and Proteus mirabilis were performed. An alignment of CAT amino acid sequences demonstrated the presence of 34 conserved amino acids among all CAT variants. These conserved residues were considered for their possible roles in the structure and function of CAT. On the basis of the alignment, a phylogenetic tree was constructed. It demonstrated relatively large evolutionary distances between the CAT variants of enteric bacteria, Clostridium, Bacillus, and Staphylococcus species.

Amino Acid Sequence