Measurement of serum gentamycin levels by rapid tests

Gentamicin is a very useful antimicrobial agent for the treatment of serious infections caused by gram-negative bacteria. However, it's low therapeutic index and potential ototoxic and nephrotoxic side effects necessitate frequent determinations of serum concentration to assist in maintaining therapeutic levels and avoiding toxic levels. Two bioassays and a latex agglutination inhibition card (LAIC) test were evaluated to determine gentamicin levels in nearly 100 patient sera. Results were compared with a radioimmunoassay (RIA). Two bioassays, the Bio-Monitor and the GentaSak, gave correlation coefficients of 0.987 and 0.982, respectively. The correlation coefficient for the LAIC test was 0.987. All three tests compared well with RIA in accurately detecting gentamicin levels in patient as well as simulated sera. The LAIC test, however, was more rapid, giving results within half an hour whereas bioassays required 6–8 hours for completion. The LAIC test was also found to be more economical. It provides a suitable alternative to RIA procedures in small laboratories and for performing stat tests since batching is not necessary.     

Synthesis of charged amphipathic nitroxide lipid spin labels and an example of their application in membrane studies

The synthesis of a series of amphipathic nitroxide lipid spin labels is reported. Thus, 12-proxylhexadecanol has been converted into the versatile fatty acid spin label 14-proxylstearic acid. This substance was used to prepare 14-proxylstearyltrimethylammonium methanesulfonate, a positively charged label, and 14-proxylstearylmethyl phosphate sodium salt, a negatively charged label. Also prepared in the doxyl series were quaternary ammonium salts derived from 16-doxyl- and 7-doxylstearic acid. The positively charged and negatively charged proxyl labels were used in a preliminary experiment to investigate the role of charge in their interaction with reconstituted cytochrome oxidase. The average binding affinity of the negatively charged label is approximately 2-fold higher than that of the positively charged label at pH 7.4. At pH 5.5 the average relative affinity for negatively charged label is about 3.5-fold higher than that of positively charged label, suggesting that the ionizable group(s) on the protein can interact with the lipid headgroup.     

Thyrotropin-releasing hormone facilitates display of reproductive behavior and locomotor behavior in an amphibian

In the amphibian brain, thyrotropin-releasing hormone (TRH) is present in many regions outside the hypothalamus. The functions of this extrahypothalamic TRH however are unknown. We sought to determine whether TRH or its metabolites altered reproductive behaviors (amplectic clasping behavior) or locomotor behaviors of the male South African clawed frog, Xenopus laevis. TRH-injected (100 micrograms; dorsal lymph sac injection) male Xenopus displayed significantly fewer amplectic clasp attempts and longer clasp durations than saline-injected controls. The TRH metabolites, TRH acid and histidylproline diketopiperazine, similarly altered clasping behavior. Several hormones released by TRH, including thyroid-stimulating hormone, melanocyte-stimulating hormone, prolactin, and dopamine, had no significant effect on clasp frequency or duration. Locomotor activity in Xenopus males was increased significantly after 15 min following TRH injection (150 micrograms); this effect persisted for at least 1 hr. The metabolites did not alter locomotion. These studies indicate that TRH can facilitate the display of two behaviors in the South African clawed frog. Effects of TRH on locomotor and reproductive behaviors thus appear in several vertebrate classes. These behavioral actions of TRH likely occur through different mechanisms or at different sites.     

Effect of vasotocin on locomotor activity in bullfrogs varies with developmental stage and sex

Although neurohypophysial peptides are present in many regions of the developing and adult bullfrog (Rana catesbeiana) brain, the function of these peptides remains unclear. To investigate possible behavioral actions, we examined locomotor activity following peptide injection in bullfrogs at various developmental stages. An intraperitoneal (ip) injection of arginine vasotocin (AVT) in tadpoles (stages V, X, or XVII) produced an immediate and dose-dependent inhibition of locomotor activity. On the other hand, AVT stimulated activity when administered ip to juvenile or adult female bullfrogs, but did not influence activity in juvenile or adult males. The minimum effective dose of AVT, when injected directly into the brain of tadpoles, was 100-fold less than that observed when injected ip, suggesting a central nervous system site of action for this peptide. A vasopressin receptor antagonist (d(CH2)5[Tyr(Me)2]AVP administered ip or icv) significantly increased locomotor activity in tadpoles, compared to controls. Oxytocin, vasopressin, and AVP4-9 inhibited activity in tadpoles while mesotocin, des Gly(NH2)AVP, and pressinoic acid had no significant effect. Injection of PGF2 alpha also significantly decreased activity levels in tadpoles. However, pretreatment of tadpoles with indomethacin, a prostaglandin synthesis inhibitor, did not prevent the behavioral effects of AVT, suggesting that prostaglandin synthesis is not required for this response. In summary, AVT influenced locomotor activity in bullfrog tadpoles and female frogs. This effect shifted during development from an inhibitory action in tadpoles to a stimulatory effect in metamorphosed female frogs. The effect of AVT on juvenile and adult frog locomotion was sexually dimorphic, as this peptide altered female behavior but not male behavior.     

Effect of synthetic enantiomeric cannabinoids on platelet aggregation.

The effect of a synthetic pair of enantiomeric cannabinoids on platelet function was evaluated. The nonpsychotropic enantiomer, the 1,1-dimethylheptyl homolog of (+)-(3S,4S)-7-hydroxy-delta-6-tetrahydrocannabinol (HU-211), was found to be more active in inhibiting ADP-induced platelet aggregation than the highly psychotropic (-)-enantiomer (HU-210). The related (+)-(3R,4R) cannabinoid, HU-213, which lacks the 7-hydroxy moiety, exerted its inhibitory effect within a wider range of concentrations. The results indicate a differentiation between psychotropic activity and inhibition of platelet aggregation in the cannabinoid group of compounds.     

Differential bioavailability of soil-sorbed naphthalene to two bacterial species.

Prediction of the fate of hydrophobic organic contaminants in soils is complicated by the competing processes of sorption and biodegradation. To test the hypothesis that sorbed naphthalene is unavailable to degradative microorganisms, we developed a simple kinetic method to examine the rates and extents of naphthalene degradation in soil-free and soil-containing systems in a comparison of two bacterial species. The method is predicated on the first-order dependence of the initial mineralization rate on the naphthalene concentration when the latter is below the Michaelis-Menten half-saturation constant (Km) for naphthalene for the organism under study. Rates and extents of mineralization were estimated by nonlinear regression analysis of data by using both a simple first-order model and a three-parameter, coupled degradation-desorption model described for the first time here. Bioavailability assays with two bacterial species (Pseudomonas putida ATCC 17484 and a gram-negative soil isolate, designated NP-Alk) gave dramatically different results. For NP-Alk, sorption limited both the rate and extent of naphthalene mineralization, in accordance with values predicted on the basis of the equilibrium aqueous-phase naphthalene concentrations. For strain 17484, both the rates and extents of naphthalene mineralization exceeded the predicted values and resulted in enhanced rates of naphthalene desorption from the soils. We conclude that there are important organism-specific properties which make generalizations regarding the bioavailability of sorbed substrates inappropriate.     

Linear order of new and established DNA markers around the fragile site at Xq27.3

We have used recombinant clones derived from microdissection of the fragile X region to characterize breakpoints around the fragile site at Xq27.3. So far, no microdissection markers derived from Xq28 material have been found, thus allowing a rapid screening for clones surrounding the fragile site by their presence in a somatic cell hybrid containing Xq27.2-Xqter. A total of 43 new DNA markers from Xq27 have been sublocalized within this chromosome band. Of these new DNA markers, 5 lie in an interval defined as containing the fragile X region. The saturation of Xq27 with DNA markers by microdissection demonstrates the power of this technique and provides the resources for generating a complete physical map of the region.     

Snow-patch foraging by pink-footed geese Anser brachyrhynchus in south Iceland

The pre-nesting feeding behaviour of pink-footed geese was studied in hayfields in southern Iceland during the late spring of 1989. Persistent snow-patches protected underlying grass from the effects of severe night-time frost. Areas within 1 m of snow-patches had significantly greater amounts of green material than those further away; green material contained more than double the protein of brown, dead material, which predominated in open fields. Geese spent nearly 60 times more time feeding within 1 m of snow patches than expected by chance, and 20 times more time within 2–5 m. Their feeding rates here were faster and their step rate slower than further away. In this way, the geese selected the prime forage as soon as it became available.     

Quantitative staging of embryonic development of the tobacco hawkmoth,Manduca sexta

Summary A complete timetable of embryonic development of the tobacco hawkmoth,Manduca sexta (Lepidoptera: Sphingidae), is presented. Using living embryos, 20 developmental stages from oviposition to hatching are described with respect to their morphological and physiological maturation. This staging series provides a simple method to identify the stage ofManduca development during all phases of embryogenesis.