A rapid automatic method for determining shock sensitivities of mice

An instrumental technique is described which permits rapid blind determination of shock sensitivities of mice. The measured parameter of shock sensitivity was not affected by increased activity of the subjects. Morphine was found to decrease the shock sensitivity as a function of dose.     

A method for determining information flow breakdown in clinical systems

This paper presents Determining Information flow Breakdown (DIB), a method for analyzing adverse events in clinical environments from the perspective of breakdowns in information flow. The larger context for DIB is that it is the first stage in a process that promotes organizational learning in response to adverse events through the design of novel IT solutions. DIB is based on the theoretical view of distributed cognition and adopts a system-wide view of failures. DIB is both a reactive and a proactive method in that it aims to locate the causes of either actual or potential adverse events by investigating all elements of the system related to a chosen aspect of patient care. It was developed and evolved using a case study approach whereby data on actual and potential adverse events in clinical environments was collected, modeled and analyzed using successive versions of the method. We provide an explanation and overview of the DIB method, and discuss our experiences of applying it in practice via a detailed example.     

A novel method for determining the sensitivity of herpes simplex virus to antiviral compounds

A rapid and sensitive assay was developed to analyse the sensitivity of wild type HSV-1 and HSV-2 isolates with respect to a battery of antiviral substances. In the viral sensitivity assay, human embryonic lung fibroblasts are incubated with the virus isolate and different concentrations of the antivirals. After 1-3 days, the cells are disrupted and analysed for HSV type 1 or 2 antigens by an enzyme-linked immunosorbent assay. Antigens corresponding to 17 plaque-forming units were detectable after 1 day of incubation. After 3 days, HSV antigens derived from less than one plaque-forming unit were measurable. The sensitivities of 22 HSV-1 and 19 HSV-2 primary isolates from untreated patients were tested against adenosine arabinoside, acyclovir, phosphonoformic acid and iododeoxyuridine. Each isolate was found to have an individual pattern of sensitivities to the different antivirals. Five isolates were judged to be relatively resistant to one or more of the drugs tested.     

Rapid method for determining the beta-lactamase-inducing potency of drugs

A rapid semiquantitative method for determining the -lactamase inducing potency of drugs was developed. Bacteria caryring a gene for inducible -lactamase expression were inoculated at a concentration of 10⁸ CFU/ml into microtiter plates for determination of MICs, which were recorded after 4 h of incubation. A suitable chromogenic -lactamase substrate was then added, and after incubation for another 3 h colour changes were monitored.     

The possible use ofTrypanosoma musculi infection in mice as a screening test for potentialTrypanosoma cruzi-active drugs

The ability of a range of trypanocidal drugs, including a number known to be active inTrypanosoma cruzi infections were tested againstTrypanosoma musculi infections in the mouse. The ability of these drugs, particularly in their ability to eliminate the cryptic phase ofT. musculi infections remaining in the kidneys, was investigated and their activity against this phase ofT. musculi largely paralleled their known activity againstT. cruzi infections. It is suggested that this could be used as a preliminary screening test for potentialT. cruzi-active drugs.     

A method for determining the effect of certain drugs on the motor reaction of animals

Summary A method is described for quantiative determination of the motor reaction of experimental animals to pain stimulation with the aid of a simple apparatus. The latter consists of a wooden wheel, on the axis of which a tachometer is located. A cage with a mouse is mounted over the wheel. The electrode hooks were connected by means of a fine conductor with an induction coil. When electric stimulation is applied, the animal tries to escape and so it turns the wheel of the tachometer. Stimulation with an automatic electrode is conducted according to the chosen scheme, every 2 or 5 sec for one or more minutes. From the number of revolutions for a definite period of time, prior to and after the use of the substance investigated, a conclusion may be drawn on the effect produced.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 54, No. 8, pp. 116–118, August, 1962     

Cell-permeable artificial zinc-finger proteins as potent antiviral drugs for human papillomaviruses

Human papillomavirus (HPV) is one of the important pharmaceutical targets because infection of the high-risk types causes invasive cervical cancer. However, effective antiviral drugs for HPV have not been developed so far. In the present study, we constructed cell-permeable artificial zinc-finger proteins (AZPs) by fusing an AZP previously generated for inhibition of HPV-18 DNA replication with a cell-penetrating peptide (CPP) as candidates for new antiviral drugs against HPV. We confirmed that these CPP-AZP fusions reduced the replication rate in transient replication assays when added to the culture medium. In particular, 250 nM CPP-AZP (designated AZP-R9) containing a 9-mer of arginine as the CPP reduced HPV-18 DNA replication to 3% of that of a control, and the 50% effective concentration (EC50) was <31 nM. Furthermore, a cytotoxicity assay revealed that the 50% inhibitory concentration (IC50) of AZP-R9 was >10 microM. Therefore, the selectivity index, defined as IC50/EC50, was >300, which is better than that of the antiviral cidofovir for HPVs. Thus, our results demonstrate that cell-permeable AZPs could serve as potent protein-based antiviral drugs.     

A rapid method for quantitation of antiviral antibodies

This paper examines the parameters necessary for the efficient measurement of anti-Theiler's murine encephalomyelitis virus (TMEV) antibodies in an affinity-dependent manner using a variation of a solid-phase particle concentration fluorescence immunoassay (PCFIA). By allowing antibody to react with fluorochrome-labelled virus in fluid phase and subsequently capturing the resulting virus-antibody complexes with anti-immunoglobulin coated polystyrene particles (fluid-phase PCFIA), the present assay allows for both greater sensitivity, specificity and preservation of conformational viral epitopes than do solid-phase immunoassays. Fluid-phase PCFIA proved to be a more rapid quantitative assay than ELISA and significantly diminished non-specific binding by both untreated and heat-inactivated normal mouse sera. This methodology also allowed us to perform competition assays and to determine the dissociation kinetics of anti-viral antibody preparations, investigations which cannot generally be performed as solid-phase immunoassays. Thus fluid-phase PCFIA is a rapid and efficient immunoassay with excellent reproducibility and great versatility.     

Humanized antibodies as potential drugs for therapeutic use

Antibodies have been used therapeutically to treat a variety of clinical conditions. The introduction of monoclonal antibodies (mAb) and, recently, engineered antibodies has greatly refined and expanded the therapeutic potential of this modality of treatment. Expanded use will depend on improvement in their efficacy (affinity and specificity), demonstration of their safety, and reduction of their immunogenicity depending on the size, suboptimal biodistribution and pharmacokinetics. To surmount these problems the molecules have to be redesigned and the basic issues of how monoclonal antibodies kill cells reinvestigated. The review will survey the literature for humanized antibodies in clinical trials and the perspective of the use of mAbs or engineered antibodies in clinical practice.     

Maturation in vitro of immature human oocytes for clinical use

Human oocyte maturation is considered as the reinitiation andcompletion of the first meiotic division from the germinal vesiclestage (prophase I) to metaphase II, and the accompanying cytoplasmicmaturation for fertilization and early embryonic development.Immature human oocytes obtained from patients undergoing gynaecologicalsurgery, or ovulation induction or having polycystic ovary syndrome(PCOS) can be matured and fertilized in vitro. To date, 80%of immature oocytes matured to metaphase II when cultured inmaturation medium supplemented with gonadotrophins and 85% ofmatured oocytes fertilized and cleaved in vitro. Following transferof these embryos, pregnancies and live births have been achieved.However, the capacity for oocyte maturation was different whenthe immature oocytes were retrieved from PCOS patients and whenthe oocytes were cryopreserved at germinal vesicle stage.     

A method of using noise as a test pattern for determining image enhancement filters

Many digitally based medical imaging systems include both reconstruction algorithms and additional image filters designed to enhance certain image features. However, the manufacturers usually consider these algorithms and filters to be proprietory information. The purpose of this note is to describe a simple procedure for determining the spatial frequency response of these proprietary enhancement filters. The technique uses image noise as a test pattern. The procedure consists of acquiring a small number of noise-only data sets (say 10) of a uniform phantom and reconstructing the images using the different filters with repeated use of the noise data sets. A straightforward analysis then yields the enhancement filter frequency responses.     

Influenza virus infection of hamsters. A model for evaluating antiviral drugs

Summary Inoculation of hamsters with influenza virus [A/PR/8/34HON 1] produces an inapparent infection which can be monitored by virus titrations of nasal washes or of homogenates prepared from trachea or lung. Antibody can be detected in the serum within 7 days following virus inoculation. Hamsters previously infected were found to be resistant to challenge with the same virus. The utility of this model for evaluating anti-influenza drugs was demonstrated with two compounds. Calcium elenolate, a virucidal agent, reduced the virus titers of nasal washes when the drug was given as nose drops near the time of virus inoculation so as to affect high drug concentrations in the nasal passages. Virazole, an inhibitor of virus replication, reduced the virus titers of the nasal washes when multiple drug treatments were given as nose drops in an effort to provide drug during the time of virus replication. The model described may provide a useful means of evaluating potential antiviral drug candidates inasmuch as the drug can be delivered directly into the nasal passages in a non-fatal influenza infection in a convenient laboratory animal.With 1 Figure