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A compact equipment package for vestibular experiments during spaceflight.

A compact measurement and stimulus equipment package for vestibular testing is described. The package is designed on a modular concept so that a customised version can be assembled for each experimental situation. Although primarily conceived for space-related research, the equipment has also been introduced successfully into the clinical diagnostic procedure. An essential function of the equipment is the recording and evaluation of eye movements. This is performed by a video-based measurement system which permits evaluation of horizontal, vertical and torsional components of eye movement. Objective testing of the vestibulo-ocular reflex in all three orthogonal planes is therefore possible. Furthermore evaluation of the otolithic function in weightlessness is made feasible by the possibility of measuring dynamic ocular counterrolling. Some applications of the equipment are described.

Equipment Design↗

A small mobilizable IncP group plasmid vector packageable into bacteriophage lambda capsids in vitro.

A mobilizable cosmid derivative of an IncP group plasmid was constructed by cloning the oriT region of RK2, a wide host-range plasmid, and the minimal DNA sequence of bacteriophage lambda required for efficient packaging in vitro. This cosmid is 13 kb in size and has unique restriction sites for EcoRI, XhoI, HindIII, and SalI. The XhoI and HindIII sites are within the kanamycin-resistance gene and the SalI site is in the tetracycline-resistance gene. This plasmid was mobilizable from an Escherichia coli donor to a number of diverse gram-negative bacteria at a frequency of 0.8 to 10 per 100 donors. This vector is one of the smallest of all wide host-range cosmids described in the literature. As part of this study, another mobilizable IncP group plasmid vector has also been constructed which, in addition to the sites listed above, has a unique BglII site, but which lacks the packager sequence.

Alcaligenes↗

Production of non-infectious human immunodeficiency virus-like particles which package specifically viral RNA.

A recombinant vector that rapidly produces large amounts of human immunodeficiency virus (HIV) virus-like particles (VLPs) was constructed. This vector lacks LTR sequences and a functional nef gene. The VLPs produced are non-infectious but similar in structure to mature, infectious HIV virions. They package specifically HIV RNAs containing appropriate signals and do not package abundant cellular mRNAs (e.g. actin). In the system described here, efficient particle production and release is decoupled from infection. Use of this VLP system offers many advantages over the study of infectious virions, permitting the expression of mutant phenotypes which interfere with virus infectivity.

Animals↗

Growth of Listeria monocytogenes Scott A, serotype 4 and competitive spoilage organisms in raw chicken packaged under modified atmospheres and in air.

The development of Listeria monocytogenes Scott A, serotype 4 and aerobic plate counts on minced raw chicken were determined independently at 4, 10 and 27 degrees C. Samples were packaged in flexible film under two modified atmospheres (one containing oxygen and one containing no oxygen) or air. The anaerobic modified atmosphere (75:25, CO2:N2) resulted in the failure of both the aerobic plate counts and L. monocytogenes to grow at all temperatures. Both the L. monocytogenes and aerobic plate counts grew in air at all temperatures. The aerobic modified atmosphere (72.5:22.5:5, CO2:N2:O2), which more closely duplicates commercial practice, inhibited the increase in aerobic plate counts by more than 4 log10 cfu/g compared to air at 4 degrees C. However, the L. monocytogenes was not affected by this atmosphere and increased in numbers by nearly 6 log10 cfu/g at 4 degrees C in 21 days. Regression analysis of the log10 growth and 95% confidence intervals showed that the differences between aerobic plate counts and L. monocytogenes in modified atmosphere were large. The ability of L. monocytogenes to grow in the aerobic modified atmosphere was not affected by level of the L. monocytogenes inoculum nor by the initial level of aerobic plate counts. These data show that modified atmosphere packaging of raw chicken (and probably other meats) can substantially inhibit the aerobic spoilage flora while allowing pathogenic L. monocytogenes to increase.

Animals↗

Quantification of microbial populations associated with the manufacture of vacuum-packaged, smoked vienna sausages.

Sources of contamination of vacuum-packaged vienna sausages by spoilage microorganisms were examined in a meat-processing plant. Microbial numbers present in the environment, on working surfaces and workers' hands and aprons were quantified by plate counting on selective media. Product line samples were taken at critical control points in the manufacturing process and analysed before and after preliminary incubation of vacuum-packaged samples at 25 degrees C for 24 h. In all samples the numbers of aerobic bacteria, Enterobacteriaceae, lactic acid bacteria and yeasts were determined by standard procedures. Contamination of sausage surfaces by lactic acid bacteria occurred as a result of the manufacturing and handling processes. The environment and specifically packers' hands, as well as working surfaces contributed to microbiological contamination of various types after removal of the peel from individual sausages. The preliminary incubation procedure allowed detection of low numbers of spoilage microorganisms.

Air Microbiology↗

Fungi causing thread mould spoilage of vacuum packaged Cheddar cheese during maturation.

Thread mould is a defect which occurs sporadically in maturing vacuum packaged Cheddar cheese, caused by the growth of fungi in folds and wrinkles of the plastic film in which the cheese is packaged. Fungi were isolated and identified from 110 Cheddar cheese blocks exhibiting typical thread mould defects. The major causative species were found to be Cladosporium cladosporioides, Penicillium commune, C. herbarum, P. glabrum and a Phoma species. Yeasts were also frequently isolated from the cheese, the majority belonging to the genus Candida. Fungal species which can cause thread mould defects were also found in the cheese factory environment, on cheesemaking equipment, in air, and in curd and whey, providing a wide range of potential sources of contamination.

Air Microbiology↗

Interaction of ionising radiation and acidulants on the growth of the microflora of a vacuum-packaged chilled meat product.

Microbiological effects of gamma irradiation dose of 2 kGy, with and without reduction of pH to 5.3-5.2, have been investigated with a vacuum-packaged, minced meat product prepared from pork and beef with spices and cereal fillings. Either glucono-delta-lactone or ascorbic acid were used as acidulants. Experimental batches were stored at 0-2 degrees C for 4 weeks. Effect of temperature abuse condition was also studied by transferring packages for one week to 10 degrees C after 2-week holding at 0-2 degrees C. The irradiation caused two decimal reduction of the aerobic viable cell count determined after incubation at room temperature and four decimal reduction in the Enterobacteriaceae count. Lactic acid bacteria appeared to be more radiation resistant and became the dominant component of the microflora during storage. Combination of pH-reduction and irradiation prevented growth of Enterobacteriaceae even at 10 degrees C incubation.

Animals↗

Taxonomy of lactic acid bacteria associated with vacuum-packaged processed meat spoilage by multivariate analysis of cellular fatty acids.

The taxonomy of lactic acid bacteria from vacuum-packaged processed meats is problematic, and atypical members of the leuconostocs and the Lactobacillus sake / curvatus group are often encountered. In order to resolve this problem the cellular fatty acid (CFA) content of 61 isolates from vacuum-packaged vienna sausages and 18 reference strains was determined by gas chromatography. The relationship between strains was derived by principal component analysis of data. The CFA profiles were highly reproducible. Although no relationships could be derived using only one or two differentiating CFAs, plots of the first two principal components based on only the six most variable CFAs allowed grouping of strains. The two genera (Leuconostoc and Lactobacillus) could not be clearly separated when analysed together, but differentiation of species within each of the genera was achieved when they were analysed independently. Examination of plots for the reference strains confirmed previously established relationships between these strains. From the plot of the Lactobacillus sake / Lactobacillus curvatus component of the study it was found that most atypical Lactobacillus sake / curvatus strains were closely related to the typical Lactobacillus sake isolates and reference strain, while the Lactobacillus curvatus strains formed an independent grouping. A small cluster of atypical strains, however, indicated that this relationship may not be true for all these strains. Among the leuconostocs only isolates of Leuconostoc mesenteroides could be clearly differentiated.

Fatty Acids↗

A report generator package for routine laboratory tests in a hospital cardiology department.

A simple report generating package is described. The package is designed for hospital computing personnel who wish to develop fast, easy access, easy-to-use report generating programs for use by non-computer specialists. Use of the system is illustrated by examples from three sections within a hospital cardiology department: nuclear cardiology, M-mode echocardiography and reporting of 24-hour ambulatory electrocardiogram tapes.

Cardiology↗

MIAMOD: a computer package to estimate chronic disease morbidity using mortality and survival data.

The MIAMOD package is for use in epidemiological analysis of chronic degenerative diseases. The package has been designed to use survival and mortality data supplied by the user, to estimate incidence and prevalence rates, to forecast future mortality and morbidity levels and to provide life tables. Estimates are derived by modelling incidence as an age-period-cohort function and by the equations relating specific cause mortality and prevalence to incidence and survival. The model parameters are evaluated by fitting mortality data by means of a weighted non-linear regression model. Numerical and statistical methods used to solve the regression and to compute standard errors, projections, and life tables are discussed. The program is written in standard FORTRAN 77 and runs on a mainframe computer. The different program units are illustrated together with the principal implementation options. An example of application is provided to show a typical output pattern.

Breast Neoplasms↗

ENZEDUC 1.0: a user-friendly software package for computation of hyperbolic enzyme kinetic data in biochemical education.

ENZEDUC 1.0 is a software package developed in Microsoft QuickBASIC v7.1 for computation of hyperbolic enzyme kinetics. The educational focus of the package is laboratory practical teaching. The program can manage a maximum of 18 substrate concentration values and 72 reaction rate values for each substrate concentration (1296 pairs of values). Samples can also be of unequal sizes. More than one reaction rate value for each substrate concentration can be handled. Real-time screen plots are obtained for the Michaelis-Menten function and the most popular linear transformations. Printer outputs are available. Hardware requirements are minimum 640 Kb RAM, EGA graphics card and 80286 processor.

Algorithms↗

A three-dimensional volume visualization package applied to stereotactic radiosurgery treatment planning.

A comprehensive software package has been developed for visualization and analysis of 3-dimensional data sets. The system offers a variety of 2- and 3-dimensional display facilities including highly realistic volume rendered images generated directly from the data set. The package has been specifically modified and successfully used for stereotactic radiosurgery treatment planning. The stereotactic coordinate transformation is determined by finding the localization frame automatically in the CT volume. Treatment arcs are specified interactively and displayed as paths on 3-dimensional anatomical surfaces. The resulting dose distribution is displayed using traditional 2-dimensional displays or as an isodose surface composited with underlying anatomy and the target volume. Dose volume histogram analysis is an integral part of the system. This paper gives an overview of volume rendering methods and describes the application of these tools to stereotactic radiosurgery treatment planning.

Brain Diseases↗

In vitro packaging of a lambda Dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1.

In this report we describe a coliphage lambda vector system for cloning endo R. EcoRI DNA fragments. This system differs significantly from those previously described in two ways. First, restricted and ligated DNA is encapsidated in vitro. Second, with increasing lambda DNA size in the range 78 to 100% that of wild-type, the efficiency of DNA encapsidation into infectious phage particles markedly increases. For lambda wild-type DNA the efficiency of in vitro packaging (10(6) to 10(7) plaques produced per microgram of added DNA) is equal to, or better than, the standard CaCl2 transfection method. The use of a Dam mutation to facilitate recognition of size classes of inserted fragments is described. Using this vector and in vitro packaging, several E. coli and phage P1 and R.EcoRI fragments were cloned.

Coliphages↗

Plasmids useable as gene-cloning vectors in an in vitro packaging by coliphage lambda: "cosmids".

A plasmid which contains a cos site of lambda and can be packaged into lambda bacteriophage particles is termed a "cosmid". Such plasmids can be used as gene cloning vectors in conjunction with an in vitro packaging system. The properties of a new series of cosmids based on the ColE1 replicon are described, including small temperature-sensitive plasmids which have lost mobilisation functions and carry no IS sequences. Amongst these plasmids are vectors for XmaI, BglII, BamHI, HindIII, PstI, KpnI, SalI and EcoRI. It is demonstrated that by using cosmids in particular size ranges these plasmids provide a high efficiency cloning system which yields essentially only hybrid clones without resort to a second selection or screening step, and without prior modification (e.g. phosphatase) treatment of the DNA. Attempts were made to optimise the cloning properties of the cosmid system. An Escherichia coli "gene bank" was obtained with an efficiency of 5 . 10(5) clones per microgram of E. coli DNA, and in which any particular unselected marker may be found in about one out of every 400 clones. It was demonstrated that deletion of mobilisation functions leads to loss of ability to form relaxation-complex without affecting copy number or segregation properties of the temperature-sensitive derivatives. The vectors are amplifiable in chloramphenicol to make up about 50% of the total cellular DNA.

Coliphages↗

Packaging of plasmid DNA containing the cohesive ends of coliphage lambda.

High yields of ColE1::Tn3-cos lambda plasmid genomes packaged in phage lambda virions (2.10(9) per ml) are produced by thermal induction of E. coli W3350 (lambda cI1857S7) lysogens carrying the plasmid DNA. The plasmid DNA is packaged in the linear form, with the right m' terminus of lambda being associated with the lambda tail.

Bacteriophage lambda↗

In vitro packaging into phage T4 particles and specific recircularization of phage lambda DNAs.

Concatemeric phage lambda imm434 DNA packaged in vitro into phage T4 particles produced plaques on a selective host. Moreover, lambda DNA containing a pBR322 derivative flanked by the lambda attL and attR sites could be specifically recircularized by excisive lambda recombination to yield the pBR322 derivative. A host deficient in generalized recombination and containing a defective lambda c Its prophage which provided Int and Xis proteins was the recipient for this plasmid derivative carried by T4. Such a T4-lambda hybrid may potentially allow almost one T4 headful of donor DNA (166 kb) to be packaged and recircularized.

Bacteriophage lambda↗

Isolation of fragments with pac function for phage P22 from phage LP7 DNA and comparison of packaging gene 3 sequences.

Three PstI DNA fragments of the P22-related Salmonella phage, LP7, have been cloned. They contain sequences recognized as pac signals by the packaging apparatus of P22. One of these fragments corresponds to the P22 DNA fragment carrying gene 3 which comprises the pac signal of phage P22. The product of gene 3, Gp3, is involved in the recognition of pac and the packaging process. Gene 3 of LP7 and most of the adjacent gene 2 have been sequenced. The pac analogous segments of the other two PstI fragments have been narrowed down by subcloning and by transduction of the resulting hybrid plasmids under recombination-defective conditions.

Amino Acid Sequence↗

VOSTORG: a package of microcomputer programs for sequence analysis and construction of phylogenetic trees.

VOSTORG is a new, versatile package of programs for the inference and presentation of phylogenetic trees, as well as an efficient tool for nucleotide (nt) and amino acid (aa) sequence analysis (sequence input, verification, alignment, construction of consensus, etc.). On appropriately equipped systems, these data can be displayed on a video monitor or printed as required. They are implemented on IBM PC/XT/AT/PS-2 or compatible computers and hardware graphic support is recommended. The package is designed to be easily handled by occasional computer users and yet it is powerful enough for experienced professionals.

Amino Acid Sequence↗