Changes in nonstructural protein 3 are associated with attenuation in avian coronavirus infectious bronchitis virus.

Full-length genome sequencing of pathogenic and attenuated (for chickens) avian coronavirus infectious bronchitis virus (IBV) strains of the same serotype was conducted to identify genetic differences between the pathotypes. Analysis of the consensus full-length genome for three different IBV serotypes (Ark, GA98, and Mass41) showed that passage in embryonated eggs, to attenuate the viruses for chickens, resulted in 34.75-43.66% of all the amino acid changes occurring in nsp 3 within a virus type, whereas changes in the spike glycoprotein, thought to be the most variable protein in IBV, ranged from 5.8 to 13.4% of all changes. The attenuated viruses did not cause any clinical signs of disease and had lower replication rates than the pathogenic viruses of the same serotype in chickens. However, both attenuated and pathogenic viruses of the same serotype replicated similarly in embryonated eggs, suggesting that mutations in nsp 3, which is involved in replication of the virus, might play an important role in the reduced replication observed in chickens leading to the attenuated phenotype.

Bovine serum--regulatory issues.

For many years users of bovine serum in the manufacture of human and veterinary biological medicinal products have relied upon USDA 9CFR to ensure the viral safety of their serum. Recently, EU regulators have formalised their position by issuing guidelines on the use of bovine serum during manufacture. Additionally, the European Pharmacopoeia has drafted a monograph on bovine serum. There is good harmonisation among the recommendations and requirements although the EU CHMP guideline calls for greater attention to be paid to the potential presence of infectious bovine polyoma virus. The EU guidelines also call for various tests to assess the effect of BVDV antibodies in the detection of BVDV. However, in response to criticisms from serum suppliers and users, the stringency of these recommendations is being relaxed. The overall viral safety of bovine serum should be subject to a risk analysis as infectious virus will invariably be present in some batches of serum but remain undetected. Other factors such as the geographical source of the serum, the efficacy of viral inactivation/removal steps and the ability of specific viruses to grow in the production cells being used, should be taken into consideration.

Gonadal radiation dose and its genetic significance in radioiodine therapy of hyperthyroidism.

Published estimates of radiation dose to the gonads from 131I therapy of Graves' disease vary widely, largely because of differences in assumptions regarding the details of iodine kinetics. The calculations described in this paper show that hyperthyroid patients treated with 10 mCi of 131I will usually receive a total radiation dose to the ovaries or testes of less than 3 rad. Several common roentgenographic diagnostic procedures may involve a greater radiation dose and a greater genetic hazard than does the usual 131I treatment for hyperthyroidism. It is important to minimize total exposure to radiation, but it seems unreasonable to deny 131I treatment for hyperthyroidism to young men and nonpregnant young women on the grounds of genetic hazard alone.

Radiation absorbed doses from iron-52, iron-55, and iron-59 used to study ferrokinetics.

Biological data obtained principally with Fe-59 citrate are used with physical data to calculate radiation absorbed doses for ionic or weak chelate forms of Fe-52, Fe-55, and Fe-59, administered by intravenous injection. Doses are calculated for normal subjects, primary hemochromatosis (also called idiopathic or hereditary hemochromatosis), pernicious anemia in relapse, iron-deficiency anemia, and polycythemia vera. The Fe-52 doses include the dose from the Mn-52m daughter generated after injection of Fe-52. Special attention has been given to the dose to the spleen, which has a relatively high concentration of RBCs and therefore of radioiron, and which varies significantly in size in both health and disease.

MIRD pamphlet no. 16: Techniques for quantitative radiopharmaceutical biodistribution data acquisition and analysis for use in human radiation dose estimates.

This report describes recommended techniques for radiopharmaceutical biodistribution data acquisition and analysis in human subjects to estimate radiation absorbed dose using the Medical Internal Radiation Dose (MIRD) schema. The document has been prepared in a format to address two audiences: individuals with a primary interest in designing clinical trials who are not experts in dosimetry and individuals with extensive experience with dosimetry-based protocols and calculational methodology. For the first group, the general concepts involved in biodistribution data acquisition are presented, with guidance provided for the number of measurements (data points) required. For those with expertise in dosimetry, highlighted sections, examples and appendices have been included to provide calculational details, as well as references, for the techniques involved. This document is intended also to serve as a guide for the investigator in choosing the appropriate methodologies when acquiring and preparing product data for review by national regulatory agencies. The emphasis is on planar imaging techniques commonly available in most nuclear medicine departments and laboratories. The measurement of the biodistribution of radiopharmaceuticals is an important aspect in calculating absorbed dose from internally deposited radionuclides. Three phases are presented: data collection, data analysis and data processing. In the first phase, data collection, the identification of source regions, the determination of their appropriate temporal sampling and the acquisition of data are discussed. In the second phase, quantitative measurement techniques involving imaging by planar scintillation camera, SPECT and PET for the calculation of activity in source regions as a function of time are discussed. In addition, nonimaging measurement techniques, including external radiation monitoring, tissue-sample counting (blood and biopsy) and excreta counting are also considered. The third phase, data processing, involves curve-fitting techniques to integrate the source time-activity curves (determining the area under these curves). For some applications, compartmental modeling procedures may be used. Last, appendices are included that provide a table of symbols and definitions, a checklist for study protocol design, example formats for quantitative imaging protocols, temporal sampling error analysis techniques and selected calculational examples. The utilization of the presented approach should aid in the standardization of protocol design for collecting kinetic data and in the calculation of absorbed dose estimates.

The MIRD perspective 1999. Medical Internal Radiation Dose Committee.

The MIRD schema is a general approach for medical internal radiation dosimetry. Although the schema has traditionally been used for organ dosimetry, it is also applicable to dosimetry at the suborgan, voxel, multicellular and cellular levels. The MIRD pamphlets that follow in this issue and in coming issues, as well as the recent monograph on cellular dosimetry, demonstrate the flexibility of this approach. Furthermore, these pamphlets provide new tools for radionuclide dosimetry applications, including the dynamic bladder model, S values for small structures within the brain (i.e., suborgan dosimetry), voxel S values for constructing three-dimensional dose distributions and dose-volume histograms and techniques for acquiring quantitative distribution and pharmacokinetic data.

MIRD pamphlet No. 17: the dosimetry of nonuniform activity distributions--radionuclide S values at the voxel level. Medical Internal Radiation Dose Committee.

The availability of quantitative three-dimensional in vivo data on radionuclide distributions within the body makes it possible to calculate the corresponding nonuniform distribution of radiation absorbed dose in body organs and tissues. This pamphlet emphasizes the utility of the MIRD schema for such calculations through the use of radionuclide S values defined at the voxel level. The use of both dose point-kernels and Monte Carlo simulation methods is also discussed. PET and SPECT imaging can provide quantitative activity data in voxels of several millimeters on edge. For smaller voxel sizes, accurate data cannot be obtained using present imaging technology. For submillimeter dimensions, autoradiographic methods may be used when tissues are obtained through biopsy or autopsy. Sample S value tabulations for five radionuclides within cubical voxels of 3 mm and 6 mm on edge are given in the appendices to this pamphlet. These S values may be used to construct three-dimensional dose profiles for nonuniform distributions of radioactivity encountered in therapeutic and diagnostic nuclear medicine. Data are also tabulated for 131I in 0.1-mm voxels for use in autoradiography. Two examples illustrating the use of voxel S values are given, followed by a discussion of the use of three-dimensional dose distributions in understanding and predicting biologic response.

Thyroid radioiodine uptakes and scans in euthyroid patients.

A substantial increase in the dietary iodide intake in many communities has caused a major increase in the total iodide pool in the body and a consequent decrease in the normal thyroid radioiodine uptake. Because of the pronounced regional variations in iodide supplementation of food, and because the effects of this additional dietary intake on radioiodine uptake are so large, a redefinition of the normal range of thyroid radioiodine uptake values in patients seen at this institution was thought to be indicated. The results of 6-hour and 24-hour thyroid radioiodine uptake studies in 44 euthyroid subjects, with scan data on the 20 who were studied with 123-I, are consistent with other reports indicating that the current normal thyroid radioiodine uptake values are lower than those accepted several years ago.

Comparison of three platelet markers for measurement of platelet survival time in healthy volunteers.

We studied mean platelet survival times in healthy volunteers with use of [51Cr]disodium chromate, 111In-oxine (in a solution of acid-citrate-dextrose [ACD] and saline), and 111In-tropolone (in ACD-plasma) as markers. Differences found between the 51Cr and 111In labels probably can be attributed to a variation in localization of the label on the cell and of renal handling of the free label after release. The mean platelet survival time with 51Cr was slightly longer than the survival time with both indium labels and showed a sex difference not seen with 111In-oxine. Protein-bound plasma 51Cr was lower than plasma 111In and remained constant throughout the study. Plasma 111In increased with time. For survival time calculations, no correction for free 51Cr is necessary, but correction should routinely be performed when 111In markers are used. Both 111In markers gave similar results with respect to platelet survival time. The somewhat more elaborate plasma labeling procedure with tropolone shows no measurable advantage over the original 111In-oxine method. 111In-tropolone labeling takes less time and maintains platelets in the physiologic environment of plasma during incubation. Biodistribution studies show no difference between the two 111In markers.

Genic amplification of the entire coding region of the HEF RNA segment of influenza C virus.

In order to provide an easy and powerful analysis of influenza C viral HEF RNA segment of a recent strain, a combination of reverse transcription and the polymerase chain reaction was used. We amplified the entire coding region of the HEF gene of a laboratory strain of virus called C/Johannesburg/1/66, widely used for binding and esterase activity studies as well as that of a strain isolated in 1991 (C/Paris/145/91) from a patient suffering from severe flu syndrome. The sequences we amplified were about 2 kilobases long. In this work, we show that the forward 'universal primer' Uni1, which has been used for influenza A and B viruses cDNA syntheses can also be used for influenza C virus. The PCR primers were designed to contain restriction sites to make the PCR products ready to be used for further purposes. A restriction analysis of the PCR products combined with analyses of all the human influenza C virus HEF gene sequences published so far permitted the design of sets of oligonucleotides which can prime PCR on cDNA of unknown influenza C virus for cloning.

Assuring the quality, safety, and efficacy of DNA vaccines.

Scientists in academia whose research is aimed at the development of a novel vaccine or approach to vaccination may not always be fully aware of the regulatory process by which a candidate vaccine becomes a licensed product. It is useful for such scientists to be aware of these processes as the development of a novel vaccine could be problematic owing to the starting material often being developed in a research laboratory under ill-defined conditions. This paper examines the regulatory process with respect to the development of a DNA vaccine. DNA vaccines present unusual safety considerations that must be addressed during preclinical safety studies, including adverse immunopathology, genotoxicity through integration into a vaccinees chromosomes, and the potential for the formation of anti-DNA antibodies.

Water sodium, urinary electrolytes, and blood pressure of adolescents.

Blood pressure measurements were made on children in their fourth year at secondary schools in parts of Scunthorpe Health District supplied with drinking water of varying sodium content. Of the 3131 children, 2740 were examined (1394 boys and 1346 girls). Boys had slightly higher systolic pressures and slightly lower diastolic pressures than the girls. There was no difference between the blood pressure distributions of children in areas supplied with water containing 105 mg/l sodium, 50 mg/l sodium, or less than 15 mg/l sodium. Small differences were found in the weights of children in these areas, and slightly more of the children in the area supplied with water containing more than 105 mg/l sodium had relatives who had been treated for hypertension. Standardisation for these factors did not show any relation between water salinity and either systolic or diastolic blood pressure. Studies of the urinary sodium, potassium, and the ratios of these to creatinine on a sample of 769 boys showed no correlation with assessments of usual sodium intake, but urinary sodium correlated well with salt and fluid intake at the meal immediately preceding examination. The relation between median blood pressure and urinary sodium concentration and lack of a clear relation with sodium creatinine ratio supports the hypothesis that it may be the ratio of salt to fluid intake rather than total dietary sodium that is relevant to the regulation of blood pressure.

The reaction of oxyhemoglobin with 1,1-diphenyl-2-picrylhydrazine and 2,2-diphenyl-1-picrylhydrazyl.

In an effort to elucidate the mechanism of the initiation reaction of the denaturation of oxyhemoglobin, I, to methemoglobin, II, by hydrazines, we have investigated by electron paramagnetic resonance, EPR, and visible spectrophotometry at 22 degrees C, pH 7.4, the reaction of I with 1,1-diphenyl-2-picrylhydrazine, III, and 2,2-diphenyl-1-picrylhydrazyl, IV, in dimethylsulphoxide/buffer and methanol/buffer mixtures, these organic solvents included at a concentration of 10 v/v% to render III and IV soluble while not causing appreciable denaturation of I. In both buffer mixtures, the results obtained were the same. For the I/III reaction mixtures, although the spectrophotometric data obtained showed denaturation to occur, there was no EPR evidence for formation of IV, contrary to expectation based on the chemical structure of III. The EPR observations on each I/IV reaction mixture showed a rapid decrease in IV signal intensity to a value that, depending on the initial reactant concentrations, was either below the detection limit or, when measurable, constant with time. The results of similar EPR measurements on analogous II/IV reaction mixtures were the same. These EPR results are compatible with the idea that IV forms a complex with the protein moiety of I and II, and show that the I/III reaction could yield IV and thus involve a one-electron transfer process.

Samarium-145: a new brachytherapy source.

A new radiation source has been produced for brachytherapy, with radiation energies slightly above those of 125I, and a T1/2 of 340 d. This source, 145Sm, is produced by neutron irradiation of 144Sm (96.5% enriched). Decay is by electron capture with 140 K x-rays per 100 disintegrations in the energy region between 38-45 keV, plus 13 gamma-rays at 61 keV. These sources are encapsulated in Ti tubes, approximately 0.8 mm X 4.5 mm, and have been developed for temporary implantation in brain and ocular tumours. The 38-61 keV photons should make such sources easy to shield, while providing a dose distribution from source arrays somewhat more homogeneous than that from 125I. In addition, the 340 d half life of 145Sm permits its use for times significantly longer than that of 60 d 125I. While the 145Sm sources have been designed primarily for implantation in a brain tumour, they should be useful for almost any conventional brachytherapy application.

Facial reanimation after acoustic neuroma excision: the patient's perspective.

Facial reanimation after acoustic neuroma excision is currently accomplished using a variety of surgical techniques. A multi-institutional survey of patient perceptions of facial reanimation success was accomplished by mailing a questionnaire to 809 randomly selected members of the Acoustic Neuroma Association. Four hundred sixty patients who underwent 296 reanimation procedures responded. Facial to hypoglossal nerve anastomosis, tarsorrhaphy, and upper eyelid implants were most frequently performed. The patient's estimations of initial deficit, spontaneous recovery, and overall satisfaction with the reanimation procedures are discussed.

Assuring the quality, safety, and efficacy of DNA vaccines.

Scientists in academia whose research is aimed at the development of a novel vaccine or approach to vaccination may not always be fully aware of the regulatory process by which a candidate vaccine becomes a licensed product. This chapter will provide an overview of the regulatory process and will discuss in more detail the quality and pre-clinical safety issues of plasmid DNA vaccines intended for human use. It is useful for research scientists to be aware of these processes as the development of a novel vaccine could be problematic due to the starting material often being developed in a research laboratory under ill-defined conditions.

Retroviral aspects of the characterisation of avian and mammalian cell substrates.

There have been no reported incidents of adventitious retroviral infection of cell lines used for the production of vaccines and other biologicals. However, due to the unique molecular biology of retroviruses, cell lines may contain endogenous retroviral genomes and these can give rise to defective retroviral particles, e.g. murine hybridomas and CHO cells. Recently, a similar situation was reported for a cell substrate in use for decades for vaccine production, namely chick embryo fibroblasts, and this raised some concern regarding the safety of these vaccines.

European Union guidance on the quality, safety and efficacy of DNA vaccines and regulatory requirements.

A European Union note for guidance on gene transfer medicinal products is being developed by the Biotechnology Working Party (BWP) of the Committee for Proprietary Medicinal Products (CPMP) which will include guidance for DNA vaccines. The 'Note for Guidance on the Quality, Preclinical and Clinical Aspects of Gene Transfer Medicinal Products' outlines the information required to assure the quality of the plasmid DNA intended to be used as a vaccine. It also provides guidance on preclinical safety evaluation and on clinical efficacy and safety evaluation with regard to obtaining marketing authorisation according to Council Regulation No. (EEC) 2309/93. Before initiating a clinical trial, it is necessary to obtain an appraisal from the relevant central and/or local ethics committees, and the competent authority within the member state(s) concerned has to authorise, or be notified of, the clinical trial. Harmonisation of gene therapy and DNA vaccine regulations as well as co-operation between relevant ethics committees and authorities in Europe is currently being improved.

[Differences in receptor specificity between the influenza A viruses isolated from the duck, chicken, and human]. / Sravnenie retseptornoi spetsifichnosti virusov grippa A, vydelennykh ot utok, kur i cheloveka.

The affinity of the duck, chicken, and human influenza viruses to the host cell sialosides was determined, and considerable distinctions between duck and chicken viruses were found. Duck viruses bind to a wide range of sialosides, including the short-stem gangliosides. Most of the chicken viruses, like human ones, lose the ability to bind these gangliosides, which strictly correlates with the appearance of carbohydrate at position 158-160. The affinity of the chicken viruses to sialoglycoconjugates of chicken intestine as well as chicken, monkey, and human respiratory epithelial cells exceeds that of the duck viruses. The human influenza viruses have high affinity to the same cells but do not bind at all to the duck epithelial cell. This testifies to the absence of 6'-sialylgalactose residues from the duck cells, in contrast to chicken and monkey cells. The alteration of the receptor specificity of chicken viruses in comparison with duck ones results in the similarity of the patterns of accessible cells for chicken and human influenza viruses. This may be the cause of the appearance of the line of H9N2 viruses from Hong Kong live bird markets with receptor specificity similar to that of H3N2 human viruses, and of the ability of H5N1 and H9N2 chicken influenza viruses to infect humans.