Rapid Determination of Salmonella in Samples of Egg Noodles, Cake Mixes, and Candies

A glass apparatus system was compared with a standard enrichment broth-selective agar method to test samples of egg noodles, cake mixes, and candy for the presence or absence of salmonellae. The glass apparatus system used fermentation of mannitol, production of H(2)S, or motility, in conjunction with a serological test of flagellar antigens, to detect salmonellae. No salmonellae were detected in 173 samples of food products. Of these samples, 171 were found to be Salmonella-negative after 48 hr with the glass apparatus system. After 72 hr, the standard Salmonella procedure yielded 38 samples which produced Salmonella false-positive results on selective agars. Inoculation of samples with cultures of Salmonella showed that approximately one inoculated cell could be detected after 48 hr of incubation with the glass apparatus. The standard Salmonella test requires a minimum of 72 hr for completion. Compared with the standard Salmonella test, the glass apparatus system is a more rapid and simple system that can be used to determine the presence or absence of Salmonella in these food products.     

An automatic gas counter for quantitative micro-determinations of tritium in biological material

An automatic microcounter for quantitative determinations of tritium has been developed. It has a background of 0.5 cpm and a constant and quench-free efficiency of 57.4 ± 2.1% ($\text{E}{}^2\text{b}$ 6600). Radioactivities of 2 pCi can be measured with a standard deviation of ±10% using a 1-hr counting time. It is particularly adapted for tritium microassay from biological material in which tritium is converted to a gaseous state by a single step combustion and reduction method, utilizing a mixture of metallic zinc and anhydrous sodium carbonate. Samples weighing up to 2 mg can be measured using sample ampules made of Pyrex glass with outer diameter of 5 mm and length of 30 mm. The material costs are extremely low and one technician can load and scal over 100 samples in a day. The apparatus may have significance as an additional instrument beside liquid scintillation counters in the region of radioactivities below 20 pCi. The method has been successfully used in investigations concerning steroid metabolism in rat adrenal cortex and testis. Specific activities have been determined from tissue samples weighing 1 μg and less.     

Synthesis of C-19 deuterium labelled steroids.

Preparation of the synthetically useful steroid intermediate 19-d3-androst-4-ene-3,17-dione (Ia) together with its 19-d2-(Ib) and 19-d1-(Ic) analogues is described. The conditions and work-up of the synthesis have been designed to eliminate tedious chromatographic separation and purification steps thus enabling decigrams of material to be conveniently prepared using standard laboratory apparatus. The deuterium label in the C-19 angular methyl group is inert to normal chemical exchange processes, thus offering the opportunity for synthesis of more complex, biologically active, stable labelled steroids, whose metabolism can be studied mass spectrometrically.     

An automatic apparatus for continuous suspension culture with a concentrating device

A chemostat in which mammalian cells can be raised in continuous suspension culture is described. It is constructed from commercially available parts. This apparatus has the advantage over earlier models in that the medium can be pumped off free of cells, thus suddenly increasing the cell concentration in the culture. The apparatus has been successfully used in studies on contact inhibition.     

Quantitative tracer studies of metabolic dynamics of animal cells growing in tissue culture

Metabolic dynamics of animal cells, growing in tissue culture, can be determined quantitatively by allowing the cells to metabolize radioactively labeled substrates under carefully controlled steady-state conditions. In order to avoid artifacts resulting from uncontrolled changes in physiological conditions, a steady-state apparatus for animal cells (SAFAC) has been constructed. In this device, cells in culture plates (max of 30) can be given radioactive substrate and incubated for various periods without disturbing the steady-state metabolism prior to killing. Subsequent analysis by two-dimensional paper chromatography and radioautography shows that metabolites are labeled rapidly and thereafter are maintined at constant levels of radioactivity, as expected for steady-state metabolism.     

Application of a novel apparatus, the quartz chemical analyzer, to the determination of endotoxin in blood.

A novel apparatus called a quartz chemical analyzer (QCA) has been developed using a quartz crystal resonator. This apparatus measures sample viscosity changes based on resonant frequency changes of the quartz crystal. The apparatus was used to determine bacterial endotoxin concentrations by monitoring the gelation reaction of Limulus amebocyte lysate. The QCA determined endotoxin concentrations with good accuracy and reproducibility in the range of 0.001-3 EU/ml for endotoxin standard (JP XII). For endotoxin determination in human whole blood and plasma samples, the inhibitory reaction was eliminated by pretreatment of a fourfold dilution at 60 degrees C and incubation for 30 min. There are many advantages of the QCA method compared with the turbidimetric and chromogenic methods. For example, QCA can measure sample viscosity changes with high sensitivity and accuracy because QCA detects minor resonant frequency changes and the frequency data give a numerical value for easy quantitation. QCA can examine turbid samples, and the required quantities of samples and reagents are small, since the quartz crystal detects sample viscosity changes directly. The endotoxin determination time may be shortened by raising the reaction temperature, and QCA can detect other types of coagulation reactions.     

一种新型半导体激光治疗机研制

采用二合一光纤系统,高效率光纤耦合装置,AlGaInp半导体激光器, 方便适用的光纤定位装置以及稳定可靠的电源控制系统,研制了一种新型650nm半导体激光治疗仪,其光纤端输出达109mW。该仪器已在生物医学领域成功应用。     

核黄素基因工程研究进展

核黄素 (维生素B2 )为天然水溶性的B族维生素 ,是维持机体正常代谢所必须的物质 ,具有重要的生理功能。目前核黄素的生产方法主要有化学合成法和微生物发酵法。其中微生物发酵法是后来发展起来的一种十分经济有效的方法 ,并在核黄素主产中开始占据主导地位。为进一步获得核黄素高产菌株 ,人们对核黄素合成基因及其表达调控的机制做了深入细致的研究 ,并以此为依据 ,通过基因工程手段构建出了核黄素高产菌株 ,大大提高了核黄素的产量 ,其中尤以枯草芽孢杆菌最为成功。综述发酵法生产核黄素的现状、核黄素生物合成的分子生物学以及基因工程研究进展 ,讨论了其进一步的发展方向。     

Pharmacogenomics of Cisplatin Sensitivity in Non-small Cell Lung Cancer

Cisplatin, a platinum-based chemotherapeutic drug, has been used for over 30 years in a wide variety of cancers with varying degrees of success. In particular, cisplatin has been used to treat late stage non-small cell lung cancer （NSCLC） as the standard of care. However, therapeutic outcomes vary from patient to patient. Considerable efforts have been invested to identify biomark- ers that can be used to predict cisplatin sensitivity in NSCLC. Here we reviewed current evidence for cisplatin sensitivity biomarkers in NSCLC. We focused on several key pathways, including nucleotide excision repair, drug transport and metabolism. Both expression and germline DNA variation were evaluated in these key pathways. Current evidence suggests that cisplatin-based treatment could be improved by the use of these biomarkers.     

A "slow" temperature jump apparatus built from a stopped-flow machine

A simple modification to a standard thermostated stopped-flow machine is described which allows it to be used as a temperature jump machine. Temperature jumps larger than 10 degrees C can be achieved in less than 150 ms which makes it useful for the range of times where conventional rapid temperature jumps are not applicable. The apparatus has a sample size of 300 microliters and can produce temperature jumps both above and below the initial temperature.     

Continuous Culture of Ruminal Microorganisms in Chemically Defined Medium: I. Design of Continuous-Culture Apparatus

An apparatus is described which has been used for successful continuous culture of the ciliates from the rumen of cattle. Automatic control of feeding rate, pH, oxidation-reduction potential, temperature, stirring rate, aeration rate, salinity, and volume of the culture is provided for, using standard commercial equipment, whenever possible. The operation of this apparatus is described.     

Creep indentation of single cells

An apparatus for creep indentation of individual adherent cells was designed, developed, and experimentally validated. The creep cytoindentation apparatus (CCA) can perform stress-controlled experiments and measure the corresponding deformation of single anchorage-dependent cells. The apparatus can resolve forces on the order of 1 nN and cellular deformations on the order of 0.1 micron. Experiments were conducted on bovine articular chondrocytes using loads on the order of 10 nN. The experimentally observed viscoelastic behavior of these cells was modeled using the punch problem and standard linear solid. The punch problem yielded a Young's modulus of 1.11 +/- 0.48 kPa. The standard linear solid model yielded an instantaneous elastic modulus of 8.00 +/- 4.41 kPa, a relaxed modulus of 1.09 +/- 0.54 kPa, an apparent viscosity of 1.50 +/- 0.92 kPa-s, and a time constant of 1.32 +/- 0.65 s. To our knowledge, this is the first time that stress-controlled indentation testing has been applied at the single cell level. This methodology represents a new tool in understanding the mechanical nature of anchorage-dependent cells and mechanotransductional pathways.     

An Apparatus (Georgiou-Petri Dish) for Growing Fungi and other Microorganisms on Liquid Media in a Petri Dish

This work describes a new apparatus for growing fungi and other microorganisms on liquid nutrient media in a Petri dish. The apparatus is composed of a net supporting a cellophane membrane stretched between an outer and an inner ring that is placed inside a Petri dish. This modification of the standard Petri dish offers many advantages for studying growth, metabolism, differentiation, and other aspects of fungi in liquid cultures with minimal waste of expensive chemicals. Monitoring of excreted or absorbed substances by the fungi, the aseptic transfer of undisturbed fungal colonies from dish to dish, and harvesting are made easier, using this apparatus.     

An airlift pump device for low pressure perfusion storage of the isolated heart

A portable apparatus for the continuous hypothermic perfusion of the isolated heart is described. The system has been used successfully to store pig and baboon hearts for periods of up to 48 hr, and to store human donor hearts for periods of 7 to 17 hr before being transplanted. The perfusate is both oxygenated and circulated by gas flow from a pressurized oxygen cylinder, using the air-lift pump principle. The apparatus has no moving parts and requires no electrical energy supply; malfunction is, therefore, extremely unlikely. A regulator has been incorporated which can be adjusted to increase or decrease the myocardial perfusion pressure. The system and environmental variables which can affect flow and pressure within the apparatus are discussed. The storage time allowed by this system will enable transportation of donor hearts between most of the world's major cities.     

Isoelectric focusing with reduced cathodic drift and migration into the anode chamber

An apparatus has been developed to reduce cathodic drift and migration into the anode chamber in vertical gel rod isoelectric focusing (IEF). In contrast to commercially available apparatuses, this apparatus can easily handle many more gels at one time, and the length, diameter and shape of its gel can be arbitrarily changed. In addition, high concentrations of detergent can be used to dissolve the protein samples, and removal of the gel cylinders from the glass tubes is easy.     

A gradient titration apparatus for determining spectrophotometric binding isotherms.

The construction of an automatic gradient titration apparatus using a multichannel peristaltic pump and a recording spectrophotometer is described. The ability of the apparatus to faithfully generate continuous spectrophotometric binding isotherms was tested in experiments studying the interaction of DNA with neutral red. The method has been shown to require low volumes of reactants, and complete binding curves can be produced in less than 15 min. The apparatus was also used to perform automatically the method of continuous variations in experiments determining the binding stoichiometry of calmagite and magnesium ion.     

Separation of apolipoprotein B species by agarose-acrylamide gel electrophoresis

Human apolipoprotein (apo) B has been recognized to exist in two different forms designated apoB-100 and apoB-48. The two apoB forms are usually separated by NaDodSO4 gel electrophoresis with a low percentage polyacrylamide gel in a tube gel apparatus. However, the matrix of this low percentage gel is relatively weak, and one can separate the two forms of apoB in a slab gel apparatus only if one utilizes a gradient polyacrylamide gel or a higher percentage polyacrylamide gel which results in a poorer separation of the protein bands. We have developed an agarose-acrylamide gel electrophoretic method to separate the two major apoB forms. The gel is a mixture of 0.5% agarose and 2% acrylamide. The agarose-acrylamide method is fast, has the advantage of being able to be used on an analytical or preparative scale in a vertical slab gel apparatus, and the gel is of sufficient strength to be used in immunoblotting and/or radioautography.     

A rapid and effecient method for the isolation of proteins from polyacrylamide cells.

A procedure is described for the rapid and efficient electrophoretic elution of protein from polyacrylamide gels which is then collected in a dialysis bag tied to the end of a tube containing the gel slices. To illustrate the method a heterogeneous preparation of alkaline phosphatase was used from which a single homogeneous component was isolated in six hours with a recovery of 86%. The eluted protein is collected in a volume which can easily be kept below 1.5 ml, thus eliminating the need for subsequent concentration. The method has also been used successfully in two other systems in which a human lung tumor-associated antigen and glycogen synthetase from yeast were isolated. Since the method utilizes a standard analytical gel electrophoresis apparatus with no modifications or accessories, it should be immediately applicable for the isolation of many different proteins from polyacrylamide gels.     

A Rapid and Efficient Method for the Isolation of Proteins from Polyacrylamide Gels

A procedure is described for the rapid and efficient electrophoretic elution of protein from polyacrylamide gels which is then collected in a dialysis bag tied to the end of a tube containing the gel slices. To illustrate the method a heterogeneous preparation of alkaline phosphatase was used from which a single homogeneous component was isolated in six hours with a recovery of 86%. The eluted protein is collected in a volume which can easily be kept below 1.5 ml, thus eliminating the need for subsequent concentration. The method has also been used successfully in two other systems in which a human lung tumor-associated antigen and glycogen synthetase from yeast were isolated. Since the method utilizes a standard analytical gel electrophoresis apparatus with no modifications or accessories, it should be immediately applicable for the isolation of many different proteins from polyacrylamide gels.     

A simple preparative polyacrylamide disc gel electrophoresis apparatus: purification of three branched-chain amino acid binding proteins from Escherichia coli

The equipment used for preparative polyacrylamide gel electrophoresis has been either difficult to construct or costly if purchased commercially. An inexpensive preparative acrylamide gel apparatus and peristaltic pump are described in this paper which are easy to use and may be constructed from readily available materials. The construction of the preparative gel apparatus requires no special machining or glass blowing.This report describes the use of the disc gel apparatus in the final purification step of three binding proteins which appear to be involved in the transport of the branched-chain amino acids in Escherichia coli. Two of these proteins have been described previously (1–4). The apparatus has also been successfully used in a number of other laboratories for the purification of a variety of other proteins (5–9).