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Colchicine inhibits GABA(A) receptors independently of microtubule depolymerization.

Colchicine is a microtubule depolymerizing agent used extensively in the study of cytoskeleton-dependent cell functions. In studying the possible functional interaction between the GABA(A) receptor and the cytoskeleton, we found that colchicine inhibits GABA(A) receptor function by mechanisms independent of microtubule depolymerization. Human GABA(A) receptor alpha1beta2gamma2L subunits were co-expressed in Xenopus oocytes and the effects of colchicine on GABA(A) receptor function was assessed using the two-electrode voltage-clamp technique. Co-application of GABA (10 microM) with colchicine (100 microM) resulted in a 59.9% inhibition of GABA-gated chloride currents. This effect was instantaneous in onset with no pre-incubation required and reversed within seconds. Other depolymerizing agents, such as nocodazole (20 microM) and vinblastine (200 microM), did not affect GABA(A) receptor function using the same co-application protocol used with colchicine. The polymerizing agent taxol (10-50 microM) did not affect colchicine inhibition of the GABA responses and did not itself alter GABA-gated chloride currents. The inhibitory effect of colchicine was present under conditions in which the oocyte microtubules had been depolymerized by cold temperature. These results indicate that colchicine inhibits the GABA(A) receptor via mechanisms unrelated to microtubule depolymerization. To further examine the inhibitory effect of colchicine on the GABA response, GABA (10-3000 microM) concentration-response curves were performed in the absence or presence of various concentrations of colchicine (30-300 microM). In the presence of colchicine, the GABA concentration-response curve was shifted to the right in a parallel fashion. A Schild plot of this data yielded a linear slope indicating that colchicine acts as a competitive antagonist at the GABA binding site. We conclude that colchicine is a competitive antagonist at the GABA(A) receptor and that studies using colchicine to examine the functional interaction between GABA(A) receptors and microtubules should be interpreted with caution.

Animals↗

The efficacy of cooling on excimer laser photorefractive keratectomy in the rabbit eye.

We cooled the surface of the cornea to reduce the thermal damage by the excimer laser ablation and referred to this method as "cooling photorefractive keratectomy" (cooling PRK). We performed conventional PRK and cooling PRK on rabbits' eyes and measured the thermal change during laser ablation. We also examined the degree of subepithelial haze and the tissues with a light microscope and an electron microscope. Normal corneal temperature of live rabbit corneas was about 32 degrees C and it rose to 41 degrees C after 300 laser shots. However, when we cooled the rabbit corneas to 24 degrees C with continuous irrigation of chilled BSS PLUS solution before laser ablation, the temperature rose only to 34 degrees C after the same ablation. Slit-lamp evaluation showed that more severe corneal haze occurred with the conventional PRK procedure. At 2 weeks postoperatively, the mean haze score was 1.77 +/- 0.87 in the conventional PRK and 0.87 +/- 0.38 in the cooling PRK (P < 0.01). Light-microscopy examination showed epithelial hyperplasia and fibroblasts, type III collagen, and heat shock protein-70 in the subepithelium of corneas with conventional PRK procedure than with cooling PRK. Under the electron microscope, more disruption of the layers of fibroblasts and collagen fibrils was observed in the conventional PRK procedure than that of the cooling PRK. In conclusion, the cooling of the corneal surface in PRK effectively reduces tissue damage related to subepithelial haze and may enable us to reduce the degree of myopic regression.

Animals↗

The evolution and state of modern technology for prostate cryosurgery.

Cryosurgery is the in situ ablation of a target tissue by application of extreme cold temperature. The ability of cryosurgery to ablate tissue is unquestioned. It is the controlled application of a cryoinjury in a manner to minimize morbidity that is problematic. Prostate cryosurgery is complicated by the proximity of the prostate to adjacent structures that are sensitive to a freeze injury, namely the urethra, rectal wall, and neurovascular bundles. Several recent technological advances have led to the development of an effective treatment protocol with acceptable morbidity. These include the advent of real-time transrectal ultrasound, cryomachines with almost instant freeze-thaw control through the use of the Joule-Thompson effect, and warming catheters to effectively preserve the integrity of the urethra and external sphincter. Further, temperature monitoring at the posterior margin of the prostate sometimes combined with an injection of saline solution into Denonvilliers fascia has reduced the occurrence of urethrorectal fistula formation to 0% to 0.5% in modern series. We review the key innovations of prostate cryosurgery that differentiate this state-of-the-art procedure from that used by early investigators to even that of the early 1990s. Potential future innovations, specifically related to image guidance of the procedure, are also addressed.

Body Temperature↗

Letter: Cot deaths.

Explore the source record for details and available documents.

Body Temperature↗

Activity of antimicrobial skin peptides from ranid frogs against Batrachochytrium dendrobatidis, the chytrid fungus associated with global amphibian declines.

Accumulating evidence suggests that a chytrid fungus, Batrachochytrium dendrobatidis, is responsible for recent declines in amphibian populations in Australia, Central America, Europe, and North America. Because the chytrid infects the keratinized epithelium of the skin, we investigated the possible role of antimicrobial peptides produced in the skin as inhibitors of infection and growth. We show here that 10 peptides representing eight families of peptides derived from North American ranid frogs can effectively inhibit growth of this chytrid. The peptides are members of the ranatuerin-1, ranatuerin-2, esculentin-1, esculentin-2, brevinin-2, temporin, palustrin-3, and ranalexin families. All the tested peptides inhibit growth of mature fungal cells at concentrations above 25 microM, and some of them inhibit at concentrations as low as 2 microM. A comparison of the sensitivity of infectious zoospores with that of mature cells showed that the zoospores are inhibited at significantly lower concentrations of peptides. To determine whether cold temperature interferes with the inhibitory effects of these peptides, we tested their effectiveness at both 22 and 10 degrees C. Although the peptides inhibit at both temperatures, they appear to be more effective against zoospores at the lower temperature. These results suggest that the ranid frogs have, within their repertoire of antimicrobial substances, a number of skin peptides that should be a deterrent to chytrid infection. This may provide some natural resistance to infection, but if environmental factors inhibit the synthesis and release of the skin peptides, the pathogen could gain the advantage.

Amino Acid Sequence↗

High-frequency jet ventilation cooling in a canine hyperthermia model.

A study was undertaken to evaluate high-frequency jet ventilation (HFJV) as an adjunct for the treatment of heatstroke. Eleven mongrel dogs were anesthetized, shaved, and externally heated until core temperature reached 43 C (109.4 F). Control animals (n = 6) were cooled passively in room air. Treatment animals (n = 5) were cooled with the addition of HFJV using a flow-interrupter, high-frequency jet ventilator with a rate of 200, a driving pressure of 40 psi, and an inspiratory to expiratory ratio of .5 to deliver room air. Temperatures were measured in brain, pulmonary artery, rectum, and subcutaneous tissue of the chest wall. Serial arterial blood gases, cardiac outputs, and mean arterial pressures were measured at baseline, maximal heating, and at .5, 1, 2, and 12 hours after heating. Animals were sacrificed 12 hours after heatstroke induction. Cooling rates for brain, pulmonary artery, and rectum achieved in the HFJV group were 1.7 times faster than those for the control group (P less than .05), although slower than previously reported cooling rates with gastric and peritoneal lavage. A higher PO2, a lower PCO2, and a more physiologic pH was maintained in the HFJV group than in the control group at 30 and 60 minutes after onset of cooling (P less than .05). Gross and histopathologic examination showed characteristic changes of heatstroke in both groups; no deleterious findings could be attributed to the use of HFJV itself. HFJV warrants clinical investigation as an adjunct for treatment of hyperthermia.

Animals↗

Selective gene expression during adaptation of muscle in response to different physiological demands.

Muscle is a very adaptable tissue in which gene expression is to a large extent influenced by physical signals. Adaptation to a different work regime is brought about by changes in fibre type and fibre cross-sectional area. We have shown both mass and phenotype are markedly altered by stretch and force production within a period as short as 4 days. This is associated with quantitative as well as qualitative changes in gene expression. The latter involves the expression of myosin heavy chain isogenes which encode different types of molecular motors. Some species of fish have exploited this and they are able to rebuild their myofibrillar systems for warm and cold temperature swimming by selective myosin gene expression. To understand how the different myosin isoform confer different contractile properties methods have been developed for cloning, sequencing and visualizing the structure of the ATPase site to explain how the molecular motors are designed. With regard to the chemical link between the physical signal and the upregulation of certain muscle genes we have cloned a new growth factor that is only expressed in muscles subjected to stretch and/or exercise and which is designed for autocrine/paracrine action. Experiments indicate that the expression of a local growth factor which induces repair, remodelling and hypertrophy is one of the ways cells respond to mechanical strain.

Adaptation, Biological↗

Effects of Erkang, a modified formulation of Chinese folk medicine Shi-Quan-Da-Bu-Tang, on mice.

Shi-Quan-Da-Bu-Tang is a traditional Chinese herbal medicine formula used to increase vital energy, and strengthen health and immunity. Data from previous studies demonstrated that this formula also has the ability to attack tumor tissue. The Erkang capsule is a modified formula of Shi-Quan-Da-Bu-Tang, with the addition of four other herbs to increase the adaptogen effects and ergogenic properties. Results from this study in mice indicated that the Erkang treated group had significant differences in mortality, body weight change, fatigue, cold temperature endurance, and immune function related organ weight change, compared to the control animals.

Animals↗

Glistening formation in an AcrySof lens initiated by spinodal decomposition of the polymer network by temperature change.

PURPOSE: To examine the effect of temperature change on the structural alteration and glistening formation in an AcrySof intraocular lens (IOL). SETTING: Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, Kobe, Japan. METHODS: The equilibrium water content of a Wagon-Wheel-packaged AcrySof IOL was determined at various temperatures by the gravimetrical method. Alterations in the microscopic structure of the lens were analyzed in situ with a reflecting microscope without staining, while the temperature of the lens was changed at different rates. RESULTS: Microvacuoles of 1.0 to 20.0 microm formed in the bulk of a swollen AcrySof IOL as the temperature decreased. The morphology of the vacuoles was similar to that of glistening particles observed in vivo. The size and density of the glistening-like vacuoles varied depending on the magnitude and rate of the temperature decrease. The glistening-like vacuoles were imprinted in the bulk of the IOL during the preceding cooling process in aqueous humor. CONCLUSIONS: Glistenings formed in the bulk of an AcrySof IOL as the medium temperature decreased. The mechanism may involve spinodal decomposition of the swollen polymer network, which initiates the formation of microvacuoles consisting of water and loosely packed network chains.

Acrylic Resins↗

Estrogens, brain and behavior: studies in fundamental neurobiology and observations related to women's health.

Mechanisms and consequences of the effects of estrogen on the brain have been studied both at the fundamental level and with therapeutic applications in mind. Estrogenic hormones binding in particular neurons in a limbic-hypothalamic system and their effects on the electrophysiology and molecular biology of medial hypothalamic neurons were central in establishing the first circuit for a mammalian behavior, the female-typical mating behavior, lordosis. Notably, the ability of estradiol to facilitate transcription from six genes whose products are important for lordosis behavior proved that hormones can turn on genes in specific neurons at specific times, with sensible behavioral consequences. The use of a gene knockout for estrogen receptor alpha (ERalpha) revealed that homozygous mutant females simply would not do lordosis behavior and instead were extremely aggressive, thus identifying a specific gene as essential for a mammalian social behavior. In dramatic contrast, ERbeta knockout females can exhibit normal lordosis behavior. With the understanding, in considerable mechanistic detail, of how the behavior is produced, now we are also studying brain mechanisms for the biologically adaptive influences which constrain reproductive behavior. With respect to cold temperatures and other environmental or metabolic circumstances which are not consistent with successful reproduction, we are interested in thyroid hormone effects in the brain. Competitive relations between two types of transcription factors - thyroid hormone receptors and estrogen receptors have the potential of subserving the blocking effects of inappropriate environmental circumstances on female reproductive behaviors. TRs can compete with ERalpha both for DNA binding to consensus and physiological EREs and for nuclear coactivators. In the presence of both TRs and ERs, in transfection studies, thyroid hormone coadministration can reduce estrogen-stimulated transcription. These competitive relations apparently have behavioral consequences, as thyroid hormones will reduce lordosis, and a TRbeta gene knockout will increase it. In sum, we not only know several genes that participate in the selective control of this sex behavior, but also, for two genes, we know the causal routes. Estrogenic hormones are also the foci of widespread attention for their potential therapeutic effects improving, for example, certain aspects of mood and cognition. The former has an efficient animal analog, demonstrated by the positive effects of estrogen in the Porsolt forced swim test. The latter almost certainly depends upon trophic actions of estrogen on several fundamental features of nerve cell survival and growth. The hypothesis is raised that the synaptic effects of estrogens are secondary to the trophic actions of this type of hormone in the nucleus and nerve cell body.

Affect↗

Interstellar matrices: the chemical composition and evolution of interstellar ices as observed by ISO.

Matrix isolation techniques have been developed in the early sixties as a tool for studying the spectroscopic properties of out of equilibrium species (atoms, radicals, ions, reactive molecules), embedded in rare gas inert matrices at low temperatures. Cold interstellar grains surfaces are able to condense out gas phase molecules, routinely observed by radioastronomy. These grain 'mantles' can be considered as 'interstellar matrices'. However, these matrices are not clean and unreactive. They are made principally of dirty ices whose composition must be determined carefully to assess the importance of the solid state chemistry that takes place in the Interstellar Medium. Infrared spectroscopy, both in astronomy and in the laboratory, is the unique tool to determine the chemical composition of these ices. Astronomical spectra can directly be compared with laboratory ones obtained using classical matrix isolation techniques. Furthermore, dedicated experiments may be undertaken to further improve the understanding of the basic physico-chemical processes that take place in cosmic ices.

Astronomical Phenomena↗

YidC is strictly required for membrane insertion of subunits a and c of the F(1)F(0)ATP synthase and SecE of the SecYEG translocase.

YidC was previously discovered to play a critical role for the insertion of the Sec-independent M13 procoat and Pf3 coat phage proteins into the Escherichia coli inner membrane. To determine whether there is an absolute requirement of YidC for membrane protein insertion of any endogenous E. coli proteins, we investigated a few representative membrane proteins. We found that membrane subunits of the F(0) sector of the F(1)F(0)ATP synthase and the SecE protein of the SecYEG translocase are highly dependent on YidC for membrane insertion, based on protease mapping and immunoblot analysis. We found that the SecE dependency on YidC for membrane insertion does not contradict the observation that depletion of YidC does not block SecYEG-dependent protein export at 37 degrees C. YidC depletion does not decrease the SecE level low enough to block export at 37 degrees C. In contrast, we found that protein export of OmpA is severely blocked at 25 degrees C when YidC is depleted, which may be due to the decreased SecE level, as a 50% decrease in the SecE levels drastically affects protein export at the cold temperature [Schatz, P. J., Bieker, K. L., Ottemann, K. M., Silhavy, T. J., and Beckwith, J. (1991) EMBO J. 10, 1749-57]. These studies reported here establish that physiological substrates of YidC include subunits of the ATP synthase and the SecYEG translocase, demonstrating that YidC plays a vital role for insertion of endogenous membrane proteins in bacteria.

Adenosine Triphosphatases↗

Epicatechin carbonyl-trapping reactions in aqueous maillard systems: Identification and structural elucidation.

Recently, our group reported via labeling experiments that epicatechin in Maillard reaction aqueous glucose-glycine model systems formed adduct reaction products with C2, C3, and C4 sugar fragments. In the current study, we investigated the identity of the sugar fragment precursors responsible for adduct generation by directly comparing the liquid chromatography-mass spectrometry properties of these reported epicatechin (EC)-sugar fragments adducts with those generated from reactions consisting of only EC and well-known Maillard-generated glucose fragments (i.e., glyoxal, glycolaldehyde, methylglyoxal, glyceraldehyde, etc.). The structural properties of an EC-methylglyoxal adduct reaction product were also analyzed by NMR. The most likely precursors for the C2, C3, and C4 sugar moiety of the EC-sugar fragment adducts were identified as glyoxal, hydroxyacetone, and erythrose, respectively. 1H NMR analysis of the EC-methylglyoxal indicated that the analyte underwent rapid conformational/constitutional exchange. Using cold temperature (-25 degrees C) two-dimensional NMR analyses (heteronuclear multiple bond coherence, heteronuclear multiple quantum coherence, and 1H-(1)H correlation spectroscopy), the structure of one of the isomers was reported to consist of a covalent linkage between the C1 position of the methylglyoxal and either the C6 or the C8 position of the EC A ring, presumably generated by hydroxyalkylation and aromatic substitution reactions.

Carbon↗