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An assessment of the microbiological risks involved with egg washing under commercial conditions.

The potential benefits of washing eggs is offset by a historical perception in the European Union that wetted eggs are prone to spoilage and water loss. This study describes the effects of spray jet washing under various processing conditions to shell surface counts of Salmonella and the presence of bacteria in egg contents. Experiments used eggs that were contaminated with Salmonella Enteritidis PT4 or Salmonella Typhimurium DT104 before cuticle hardening. Washing of contaminated eggs under optimum conditions resulted in a more than 5-log reduction of Salmonella counts from the shell surface. Salmonella was not isolated from the yolk or albumen of any egg washed by the optimal protocol, suggesting that when properly controlled, egg washing did not cause Salmonella to enter the contents. However, contamination did arise if strict control was not maintained over the wash and rinse water temperatures. Both Salmonella Enteritidis and Salmonella Typhimurium were shown to enter the egg contents when water temperatures were lowered, indicating that strict temperature control must be maintained in order to prevent the ingress of Salmonella into egg contents. Other washing machine parameters that were investigated did not significantly affect Salmonella entry into the egg contents but influenced shell surface kill levels to varying degrees.

Animals↗

Iron supplementation in adolescent hemodialysis patients.

AIMS: Iron supplementation is necessary in children on hemodialysis, but the optimal protocol remains unknown. We studied the effects of changing our unit's protocol from oral iron with periodic doses of parenteral iron dextran to routine administration of parenteral sodium ferric gluconate on anemia and iron parameters. METHODS: We followed seven hemodialysis patients aged 15 20 years (mean 17 years). Hemoglobin, hematocrit, serum iron, transferrin saturation, ferritin, erythropoietin dose, total elemental iron dose and total iron cost for the six months prior to the protocol change were compared to the same variables during the six months following the change. RESULTS: There was no statistically significant difference between the doses of parenteral iron between the two protocols; however, the total dose of elemental iron administered in the oral iron plus iron dextran protocol was greater than in the sodium ferric gluconate protocol (19.6+/-13.1 (mean+/-SD) mg/kg/week versus 1.1+/-0.3 mg/kg/week; p = 0.03). Both protocols had equivalent efficacy with respect to hemoglobin, ferritin and other measures of iron stores. On the other hand, the costs of sodium ferric gluconate were significantly higher than those of oral iron plus intermittent iron dextran (157.75+/-45.94 $/patient/month versus 52.08+/-49.88 $/patient/month; p = 0.01). CONCLUSIONS: Routine administration of sodium ferric gluconate is equivalent if not superior to use of oral iron plus iron dextran for maintenance of iron stores in adolescents on hemodialysis, but more expensive.

Administration, Oral↗

Use of buccal cells collected in mouthwash as a source of DNA for clinical testing.

CONTEXT: To maximize the participation rate in population genetic studies, alternatives to invasive whole blood collection are increasing. One such alternative is buccal epithelial cell collection, which, in contrast to venipuncture and finger sticks, is painless. Buccal cells, if collected and purified efficiently, offer an acceptable source for DNA to be used in research and clinical applications. OBJECTIVE: To develop a noninvasive sampling method for collecting cells for routine DNA testing in a clinical laboratory setting. DESIGN: Five factors were used to evaluate several brands of mouthwash: (1) compatibility with the DNA purification chemistry, (2) DNA yield, (3) DNA quality, (4) DNA stability at room temperature, and (5) mouthwash taste. Next, an optimization study was undertaken to maximize DNA yield. Finally, a validation study was undertaken with the optimized protocol to test a panel of 14 donors for DNA yield and performance and to test for the stability of DNA held in mouthwash. SETTING: Industrial research and development laboratory. RESULTS: Of 5 mouthwashes tested, Scope brand mouthwash received the highest overall ranking. The addition of proteinase K and glycogen to the protocol significantly enhanced DNA yields, with a test panel (n = 14) giving a range of 12 to 60 microg of DNA per donor. In a 4-week room temperature stability study, the DNA in mouthwash samples was found to be stable for at least 2 weeks. CONCLUSION: A clinically validated DNA purification chemistry was adapted to a noninvasive specimen collection method. This method used a commercially available mouthwash, Scope, to collect buccal epithelial cells for the preparation of high-quality DNA in high yield.

Cheek↗

[Shielding effect of protective seats during CT examination].

Because the exposure to radiation from CT scanning is higher than that from other X-ray diagnostic devices, it is necessary to lower this exposure. In this study, we placed a protective seat on the abdomen and dorsal side of a human body phantom. Three different kinds of CT units, with a single detector, four detectors, and 16 detectors, were used to compare the absorbed dose on the skin surface and uterus. Head and chest CT scans were taken with the standard protocol. The difference in exposure to the uterus with and without the protective shield was found to be small. Exposure to the skin surface was about 0.2 mGy without the protective seat. We found that the exposure dose to chest could be reduced as much as 50% by using the protective seat. Effective radiation differs depending on the institution, and these differences can be more than the amount that can be reduced by using the protective seat. Thus, we conclude that it is important for each institution to determine the best protocol optimization for each individual.

Female↗

[Evolution of ovarian stimulation in in vitro fertilization].

This paper reviews the different treatments of ovarian stimulation for in vitro fertilization (IVF). It emphasizes the recent development of new molecules, including recombinant gonadotrophins and GnRH antagonists. Because of their higher purity, recombinant gonadotrophins are theoretically less immunogenic than the urinary forms. Their preparation mode ensures a better batch to batch consistency. Their clinical use in IVF seems to be associated with a higher number of developing follicules and retrieved oocytes, but pregnancy rates are comparable to those obtained after ovarian stimulation with urinary gonadotrophins. The GnRH antagonists induce a direct inhibition of the pituitary gonadotrophin secretion. In association with an ovarian stimulation with gonadotrophins, they efficiently prevent premature LH surges when administered during the end of the follicular phase. The GnRH antagonists are still in clinical evaluation phase and the optimal protocol and dose still to be defined. They should become available in the coming year. The risks related to the controlled hyperstimulation together with the reduction of the number of transferred embryos has recently led "soft stimulation" to a reevaluation of the stimulation protocols towards a simplified procedure. However, the validity of this attitude could be in opposition with the wish to limit the number of transferred embryos since embryo quality seems to be directly related to the total number of embryos.

Clomiphene↗

[Comparative effectiveness of the use of hematological methods to analyze the procoagulant activity of biological materials].

Whether clinical hematological methods by employing two parameters: (plasma recalcification time (PRT) and activated partial thromboplastin time (ACTT) can be used to examine the procoagulant activity of the surface of biological materials was comparatively studied. The method PRT was shown to be sensitive and reproducible while ACTT failed to significantly record differences in the procoagulant activities of various materials. The authors chose the optimal protocol for evaluating the blood compatibility of biological materials: the surface area/plasma volume ratio is more than 150 cm2/ml, the incubation time of samples is no less than 180 sec at 37 degrees C and continuous mixing.

Biocompatible Materials↗

[Diagnostic evaluation of the breast using PET: optimization of data acquisition and postprocessing].

PURPOSE: Development and evaluation of an optimized protocol for PET examinations of the female breast with 2-F-18-fluoro-2-deoxyglucose (F-18-FDG). METHODS: All PET measurements were performed with a whole-body PET system (ECAT EXACT HR+). In order to examine the women with the breasts freely pendant, a special extension for the patient table made of carbon layer composite was designed. After data acquisition in the 3D modus, emission data were sorted into 2D sinograms using the Fourier rebinning algorithm and reconstructed by means of an ultra-fast iterative 2D algorithm (HOSP). The reconstructed emission scans were superimposed onto the corresponding transmission images. The protocol presented was evaluated in examinations on 6 women with breast lesions after the administration of 150-220 MBq F-18-FDG. From two adjacent bed positions, emission and transmission data were acquired over periods of 20 min and 10 min, respectively. For comparison, dynamic magnetic resonance (MR) image series were acquired with a whole-body MR system (MAGNETOM SP 4000) using a double-breast coil. RESULTS AND CONCLUSION: Using the designed extension of the patient table, it was possible to examine corpulent women despite the limited patient port of the PET system in the prone position with the breasts freely pendant. Alongside a reduction in motion artifacts, this positioning also offers the possibility of making a direct comparison between PET and MR images. Despite the fact that the amount of F-18-FDG applied to the patient was markedly reduced, the combination of 3D data acquisition and iterative image reconstruction resulted in excellent quality of the emission scans. By super-positioning of iteratively reconstructed emission and transmission scans, anatomical localization of breast lesions visualized on the emission scans could be improved. The postprocessing of the PET data described was completed in 60 min, this meaning that the presented concept can readily be employed in clinical practice.

Algorithms↗

[Investigation of fetal cells isolated from maternal blood by different methods].

This paper presents the results of investigation of the isolated from maternal blood by 4 different methods according to the optimized protocols. All women had male fetuses. The mononuclear cells with fetal erythroblasts were preisolated by using density-gradient centrifugation of the maternal blood in the Ficoll solution. Fetal cells were detected by FISH for Y-chromosomal sequences. The fetal cells were 2.1% by fluorescence activated cell sorting (FACS); 3.8% by magnetic field sorting, and 2.6% by two-stage density gradient precipitation. The fetal lymphocytes were investigated through air-cultivation of peripheral maternal lymphocytes. Their proportion was 3.9% in the culture samples. The findings lead to the conclusion that the new non-invasive approach is useful for prenatal diagnosis of chromosomal aneuploidies.

Aneuploidy↗

Absence of thyroid stunning after diagnostic whole-body scanning with 185 MBq 131I.

UNLABELLED: There has been recent controversy regarding the optimal protocol for imaging and ablation of post-thyroidectomy patients. Several authors have suggested that a scanning dose of 185-370 MBq (5-10 mCi) (131)I may be capable of producing a stunning effect on thyroid tissue that may interfere with the uptake and efficacy of the subsequent ablation dose of radioiodine. The purpose of this study was to determine whether a 185-MBq (5 mCi) diagnostic dose of (131)I produces a visually apparent stunning effect 72 h before (131)I ablation therapy. METHODS: One hundred twenty-two consecutive post-thyroidectomy patients for differentiated thyroid carcinoma received a 185-MBq (5 mCi) diagnostic dose of (131)I followed by a whole-body diagnostic scan at 72 h. On the same day the diagnostic scan was completed, the patient was admitted to the hospital and received an (131)I ablation therapy dose of 5550 MBq (150 mCi) in most cases. A postablation, whole-body scan was obtained at 72 h and compared with the previous diagnostic scan for any visual evidence of stunning. RESULTS: No cases of visually apparent thyroid stunning were observed on any of the postablation scans with regard to the number of (131)I foci identified or the relative intensity of (131)I uptake seen. CONCLUSION: Diagnostic whole-body scanning can be performed effectively with a 185-MBq (5 mCi) dose of (131)I 72 h before radioiodine ablation without concern for thyroid stunning.

Adolescent↗

[Significance and features of MLC reaction in histocompatibility determination of donor and recipient in allogenic bone marrow transplantation to patients with hematological malignancies].

AIM: To characterize mixed lymphocyte culture (MLC) reaction used for determination of donor-recipient compatibility before bone marrow transplantation to patients with hematological malignancies and to assess the reaction significance. MATERIAL AND METHODS: The analysis was made of compatibility testing in MLC standard reaction performed in 134 patients with hematological malignancies with HLA-A, -B identical donors-sibs and in 5 patients with haploid identical donors. RESULTS: Out of 134 patients, 22(91%) appeared compatible to donor sibs in MLC reaction, 12(9%) patients were incompatible. Mean RR for MLC-compatible couples made up: in RvD direction 77 +/- 0.17%, DvR 2.61 +/- 0.32%. 93% of RR values ranged from +15 to -15%, the rest--from +25% to -25%. Bone marrow transplantation was made in 83 patients. Graft retention was observed in 77(93%) patients. Acute and chronic graft versus host reaction developed in 15 and 17 patients, respectively. CONCLUSION: An optimal protocol is proposed for examination of compatibility donor-recipient in MLC reaction in patients with hematological malignancies. It is intended for allogenic bone marrow transplantation in hematological departments.

Adolescent↗

Technical developments in cardiac MRI: 2000 update.

In recent years, the technological developments in MRI have made it possible to perform routine cardiovascular imaging with MR. The availability of a vectorcardiogram, real time interactive scanning, SENSE and optimized protocols (balanced FFE) have made it possible to evaluate wall motion, perfusion and viability at rest and under stress. All vessels can be visualized, including the coronary arteries. In the near future, MRI will be capable of distinguishing between calcified and lipid plaque, and will even allow interventional applications.

Cardiovascular Diseases↗

Genetic alterations in tp53 in recurrent urothelial cancer: a longitudinal study.

PURPOSE: Because bladder cancer has a recurrence rate that can be as high as 90% at 2 years, we sought to clarify whether these metachronous tumors are polyclonal or monoclonal in origin. We have examined the genetic alterations of the TP53 gene in a cohort of patients with urothelial cancer who underwent multiple biopsies at different times and sites because of tumor recurrence and/or progression. We postulated that if tumor cells at different points in the natural history of the disease contain an identical mutation in the TP53 gene, this pattern could provide evidence for the monoclonality of the recurrent bladder tumors. EXPERIMENTAL DESIGN: Fifty-three biopsy specimens from 13 patients at different times and sites were selected for this study. Microdissection was used to ensure the purity of tumor cells. DNA extraction, PCR, and direct sequencing of exons 5 through 8 of the TP53 gene were conducted following protocols optimized in our laboratory. RESULTS: We found that specimens from seven patients carried tumor-specific TP53 mutations. The number of lesions in these patients ranged from two to seven, extending from 2 to 4 years. All of the seven patients displayed identical mutations in the different microdissected tumors. CONCLUSIONS: On the basis of these data, it appears that the recurrent bladder tumors originate from the same clone.

DNA Mutational Analysis↗

[Human factor VIII expression in retrovirus vector-transduced bone marrow stromal cells].

OBJECTIVE: To explore the feasibility of bone marrow stromal cells (BMSCs) used for gene therapy of hemophilia A. METHODS: Murine and rabbit BMSCs were transduced with a recombinant replication-incompetent retrovirus LNC-FVIII delta B, which encloses a B-domain-deleted (760aa-1,639aa) human factor VIII cDNA. An optimized protocol involving temperature declining (32 degrees C) and centrifugation was used. Antigen and coagulant activity of human FVIII in the cell culture medium were measured by ELISA assay and one-stage method, respectively. Semi-quantitative PCR was performed for the transduction efficiencies. Western blot was used for FVIII protein analysis. RESULTS: Transduction efficiencies were increased 5- to 8-fold by temperature decline and centrifugation, which resulted in high expression and secretion of FVIII in transduced mouse[(556 +/- 80) ng/10(6) cells/24 hr] and rabbit [480 +/- 56) ng/10(6) cells/24 hr] BMSCs. Coagulant activities of human FVIII secreted from transduced murine and rabbit BMSCs were 2.42 U/10(6) cells/24 hr and 1.8 U/10(6) cells/24 hr, respectively. Western blot analysis demonstrated that the B domain-deleted FVIII cDNA was secreted predominantly as heavy and light chain heterodimers that resemble the native forms in human plasma. CONCLUSIONS: It was demonstrated that the BMSCs system has potential utility in the gene therapy for hemophilia A.

Animals↗

[The effect of G-loads on gravity tolerance of monkeys Macaca rhesus during antiorthostatic hypokinesia].

Reported are results of 200 centrifuge runs of 24 monkeys Macaca mulatta. Microgravity effects were simulated by 4-wk immobilization with the head tilted down at -6 degrees. The investigation was divided into two series. During series-1, one group of animals was not exposed to centrifugal training, whereas the other group was exposed to +Gz at the magnitudes of 1.2, 1.4 and 1.6 G for 30-40 min 4 to 5 times a week. In series-2, again one group of animals was not trained on the centrifuge but the other was trained at +1.2 Gz for 30 min 2 or 3 times a week. Functional testing by 30-s runs at +3 Gz was performed before and after head-down immobilization. Deterioration of g-load tolerance following simulated microgravity in both groups of untrained macaques was ascribed to the deconditioning effect of simulated microgravity on the body systems. Centrifugal training had, as a rule, positive effect on g-tolerance of the animals though it failed to fully eliminate the negative consequences of simulated microgravity. This means that search after optimal protocols of application of artificial gravity generated by centrifuge needs to be continued. Evaluation of the effectiveness of combining centrifugation with other countermeasures against the adverse effects of microgravity and development of associated protocols can be another area of future research.

Adaptation, Physiological↗

Eradication of initial Pseudomonas aeruginosa colonization in patients with cystic fibrosis.

The optimal treatment for the eradication of initial P. aeruginosa infection in CF is still unclear. Recently long-term inhaled tobramycin has been proposed. Here we report the results with brief inhaled and/or systemic anti-pseudomonal treatments. Initial P. aeruginosa colonization was successfully eradicated as demonstrated by negative repetitive throat cultures or sputa and serum antipseudomonal antibodies in 15 of 17 patients for at least two years. Randomized, controlled trials are urgently needed to define the optimal protocol for the eradication of P. aeruginosa. Pseudomonas aeruginosa, infection, treatment, antibodies; inhaled tobramycin

Administration, Inhalation↗

Bronchopulmonary hyperreactivity and lung eosinophil sequestration but not their migration to the alveolar compartment are independent of interleukin-5 in allergic mice.

IL-5 is present in the lung and in the circulation following allergenic challenges in humans and in animals, but its role in bronchopulmonary hyperreactivity (BHR) and lung and bronchoalveolar lavage fluid (BALF) eosinophilia remains unclear. Because compartmentalization of IL-5 is recognized, the anti-IL-5 monoclonal antibody TRFK-5 or its isotype control GL113 were delivered selectively intranasally (i.n.) and/or intravenously (i.v.) before the prior i.n. challenge with 10 mug OVA in BALB/c and BP2 "Biozzi" mice immunized according to optimized protocols with read-outs taken 24 h later. IL-5 in the BALF was suppressed by i.n. TRFK-5, whereas its production persisted in the serum. Conversely, i.v. TRFK-5 suppressed IL-5 in the serum but not in the BALF. IL-5 was suppressed in conditioned medium from lung explants from mice treated with i.n. TRFK-5, which did not affect the other Th2 cytokines, IL-4 and IL-13. IL-5 is thus present in the alveolar, pulmonary and circulatory compartments following an i.n. allergenic challenge. When specific anti-IL-5 antibodies were delivered by the same i.n. route, BALF eosinophilia was markedly reduced, whereas BHR and lung eosinophil sequestration persisted totally or mostly, in both strains. The passage of eosinophils from lungs to alveoli depends on IL-5 released into the BALF, but not into circulation, whereas their lung sequestration and BHR are mostly IL-5-independent. IL-5 alone does not account for the complexities of BHR or of eosinophil tissue trapping, and lung-targeted immunobiologicals should be delivered into the appropriate compartment in order to assess the role of specific mediators in experimental airways/lung allergy.

Animals↗

Neonatal rat cardiomyocytes--a model for the study of morphological, biochemical and electrophysiological characteristics of the heart.

The neonatal rat cardiomyocyte model enables heart researchers to study and understand the morphological, biochemical and electrophysiological characteristics of the heart. This model offers a broad spectrum of experiments, such as studies of contraction, ischaemia, hypoxia and the toxicity of various compounds. This review examines the methodology for the isolation and cultivation of primary neonatal rat cardiomyocyte cultures, considers possible problems and pitfalls, and presents an optimized protocol for cardiomyocyte preparation.

Animals↗

New directions in photodynamic therapy.

Photodynamic therapy (PDT), a treatment approach that makes use of a photosensitizer to generate a localized toxic species in diseased tissue, has recently become an approved treatment modality. So far, however, only a handful of photosensitizers have received regulatory approval and for a small number of diseases. This chapter outlines the major limitations of PDT and speculates on the possible improvements that are required in order to advance PDT to a front line therapy. Seven areas of improvements are discussed: drug selectivity, drug delivery, light delivery, combination therapies, pigmented tumors, other potential uses, and protocol optimization. For each area, current limitations are discussed, and further required studies are recommended.

Animals↗