Sonoluminescence as an indicator of cell membrane disruption by acoustic cavitation.

Ultrasound (US) has been shown to transiently disrupt cell membranes and, thereby, facilitate the loading of drugs and genes into viable cells. Because these effects are believed to be mediated by cavitation, we hypothesized that measured levels of cavitation-induced sonoluminescence should correlate with levels of US bioeffects. We, therefore, quantified the number of calcein molecules delivered and the loss of viability in prostate cancer cells exposed to 24-kHz US over a range of different pulse lengths (1 to 100 ms), total exposure times (0.1 to 10 s) and pressures (1.0 to 9.8 atm). Consistent with previous observations, uptake increased and viability decreased with increasing pulse length, total exposure time and pressure. As a new observation, we established correlations between the amount of light produced by sonoluminescence and both molecular uptake and cell viability. These results support a cavitation-based mechanism for these bioeffects and suggest a means to control US effects on cells using sonoluminescence-based feedback.

Severe local hypothermia from laparoscopic gas evaporative jet cooling: a mechanism to explain clinical observations.

BACKGROUND AND OBJECTIVES: Explanations for laparoscopic-induced hypothermia fail to explain clinical observations. It is possible that water evaporation occurs from the jet stream of gas inflation resulting in tissue surface super-cooling leading to tissue damage and drying. METHODS: Theoretical calculations based on thermal conductivity, mass transfer effects and heat flux considerations correlated closely with synthetic and tissue experiments. Thermocouple measurements at a rate of 15 data points per second were performed. RESULTS: Cooling rates of 10 to 25 degrees centigrade per second for high flow rates were found based on gas flow rate and effective size of gas delivery site. These rapid temperature drops extended beyond a 2 cm2 diameter. CONCLUSIONS: Evaporative cooling accounts for significant hypothermia. The cooling is dependent on the lack of water vapor in the gases currently used during laparoscopy. Cooling rates are independent of height from tissue and geometry of delivery port. Heating and hydrating the gas to a physiologic condition eliminates hypothermia and tissue dessication.

What can toxins tell us for drug discovery?

Toxins are of interest in drug design because the toxins provide three-dimensional templates for creating small molecular mimics with interesting pharmacological properties. Toxins are also useful in drug discovery because they can be used as pharmacological tools to uncover potential therapeutic targets. With their high potency and selectivity, toxins are often more useful in functional experiments than standard pharmacological agents. We have used two groups of neurotoxins, the dendrotoxins and the muscarinic toxins (MTs), to explore the involvement of subtypes of potassium ion channels and muscarinic receptors, respectively, in processes involved in cognition and the changes in neuronal properties with aging. From our current work, quantitative autoradiographic studies with radiolabelled dendrotoxins reveal widespread distribution of binding sites throughout rat brain sections, but few differences exist between young adult and aged rats. However, displacement studies with toxin K, which preferentially binds to the Kv1.1 subtype of cloned potassium channel, show the selective loss of such sites in regions of the hippocampus and septohippocampal pathway with aging. MTs have been tested for effects on performance of rats in memory paradigms. MT2, which activates m1 receptors, improves performance of rats in a step-down inhibitory avoidance test, whereas MT3, which blocks m4 receptors, decreases performance when given into the hippocampus. This is the first clear demonstration of a role for m4 muscarinic receptors in cognition.

Evidence for regulation of the autoimmune anti-erythrocyte response by idiotype-specific suppressor T cells in NZB mice.

The present study demonstrates that specific CD8+, CD4- suppressor T cells (Ts) actively regulate the autoimmune anti-mouse red blood cell (MRBC) antibody response in spleen cell populations of young, Coombs-negative NZB mice. These Ts appear to bind a monoclonal NZB autoantibody (G-8 mAb) to unmodified MRBC which expresses a dominant idiotype (Id) in the spontaneous anti-MRBC autoantibody response of NZB mice. Treatment of normally nonauto-responsive spleen cells from young NZB mice with the G-8 mAb + C prior to culture allows these cells to develop, in 4-5 days, an autoantibody response to MRBC. The level of response obtained after depletion of the G-8-binding cells is comparable with that obtained after generalized depletion of Ts by treatment with anti-CD8 + C, suggesting that the G-8-binding cells make up a major portion of the regulatory Ts in this response. Yet, G-8 + C treatment depletes a very small subset of cells and not the total CD8+ T cell population. The regulatory cells appear to be neither isotype nor allotype specific, nor do they appear to have MRBC antigens bound to or expressed on their membranes. Rather, these cells are more likely G-8 idiotype specific. The regulatory G-8-binding cells are CD8+ T cells, not B cells. Furthermore, Ts-enriched populations when depleted of G-8-binding cells lose their ability to suppress in vitro anti-MRBC responses of spleen cells from Coombs-negative NZB mice depleted of CD8+ cells, as well as those of unfractionated spleen cells from Coombs-positive NZB mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Oxidative activation of Bacillus cereus spores.

A study was made of the activation of Bacillus cereus strain T spores by using the oxidizing agent sodium perborate. The degree of activation was measured with constant germination conditions by using L-alanine, inosine, adenosine, and L-alanine plus adenosine as germination stimulants. The germinal response following the various treatments was compared with the responses obtained with heat activation. It was concluded that the optimal time for activation with 30 mM sodium perborate at room temperature was about 4 hr. If the exposure time was greatly extended, the spores would germinate spontaneously. When the perborate treatment followed heat activation, the germinal response to L-alanine was stimulated, to inosine retarded and without apparent effect for adenosine or L-alanine plus adenosine. Results of experiments designed to demonstrate deactivation by slow oxidation showed that spores activated with sodium perborate were not deactivated by slow oxidation, whereas those activated by heat were. A deactivation study using mercaptoethanol as the deactivation agent showed that both methods of activation could be deactivated after a 24-hr exposure, but this deactivation was reversible by extending the exposure to mercaptoethanol. The results of heat-sensitivity studies revealed that about 70% of the sodium perborate-activated spores were heat sensitive after 60 min in a germination menstruum of L-alanine plus adenosine, whereas similarly treated heat-activated and nonactivated spores were about 99.99% heat sensitive, respectively.