Routine prophylactic antibiotic use in the management of snakebite

Background  

Routine antibiotic prophylaxis following snakebite is not recommended but evidence suggests that it may be common practice in Zimbabwe. This study set out to determine and describe the extent of this practice at Parirenyatwa Hospital, a large teaching hospital in Zimbabwe     

The "five-sites" rule and the evolution of red and green color vision in mammals

Amino acid changes S180A (S-->A at site 180), H197Y, Y277F, T285A, and A308S are known to shift the maximum wavelength of absorption (lambda max) of red and green visual pigments toward blue, essentially in an additive fashion. To test the generality of this "five-sites" rule, we have determined the partial amino acid sequences of red and green pigments from five mammalian orders (Artiodactyla, Carnivora, Lagomorpha, Perissodactyla, and Rodentia). The result suggests that cat (Felis catus), dog (Canis familiaris), and goat (Capra hircus) pigments all with AHYTA at the five critical sites have lambda max values of approximately 530 nm, whereas rat (Rattus norvegicus) pigment with AYYTS has a lambda max value of approximately 510 nm, which is accurately predicted by the five-sites rule. However, the observed lambda max values of the orthologous pigments of European rabbit (Oryctolagus cuniculus), white-tailed deer (Odocoileus virginianus), gray squirrel (Sciurus carolinensis), and guinea pig (Cavia procellus) are consistently more than 10 nm higher than the predicted values, suggesting the existence of additional molecular mechanisms for red and green color vision. The inferred amino acid sequences of ancestral organisms suggest that the extant mammalian red and green pigments appear to have evolved from a single ancestral green-red hybrid pigment by directed amino acid substitutions.      

Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae

Various selection procedures in chemostats and batch cultures were systematically tested for their efficiency to select for a multiple-stress resistance phenotype in Saccharomyces cerevisiae. To determine the relative stress resistance phenotypes, mutant populations harvested at different time points and randomly chosen clones from selected populations were grown in batch cultures and exposed to oxidative, freezing-thawing, high-temperature and ethanol stress. For this purpose, we developed a high-throughput procedure in 96-well plates combined with a most-probable-number assay. Among all chemostat and batch selection strategies tested, the best selection strategy to obtain highly improved multiple-stress-resistant yeast was found to be batch selection for freezing-thawing stress. The final mutant populations selected for this particular stress were not only significantly improved in freezing-thawing stress resistance, but also in other stress resistances. The best isolated clone from these populations exhibited 102-, 89-, 62-, and 1429-fold increased resistance to freezing-thawing, temperature, ethanol, and oxidative stress, respectively. General selection guidelines for improving multiple-stress resistance in S. cerevisiae are presented and discussed.     

Cytogenetic analyses of eight species in the genus <Emphasis Type="Italic">Leptodactylus</Emphasis> Fitzinger, 1843 (Amphibia,Anura, Leptodactylidae), including a new diploid number and a karyotype with multiple translocations

Background

The karyotypes of Leptodactylus species usually consist of 22 bi-armed chromosomes, but morphological variations in some chromosomes and even differences in the 2n have been reported. To better understand the mechanisms responsible for these differences, eight species were analysed using classical and molecular cytogenetic techniques, including replication banding with BrdU incorporation.

Results

Distinct chromosome numbers were found: 2n = 22 in Leptodactylus chaquensis, L. labyrinthicus, L. pentadactylus, L. petersii, L. podicipinus, and L. rhodomystax; 2n = 20 in Leptodactylus sp. (aff. podicipinus); and 2n = 24 in L. marmoratus. Among the species with 2n = 22, only three had the same basic karyotype. Leptodactylus pentadactylus presented multiple translocations, L. petersii displayed chromosome morphological discrepancy, and L. podicipinus had four pairs of telocentric chromosomes. Replication banding was crucial for characterising this variability and for explaining the reduced 2n in Leptodactylus sp. (aff. podicipinus). Leptodactylus marmoratus had few chromosomes with a similar banding patterns to the 2n = 22 karyotypes. The majority of the species presented a single NOR-bearing pair, which was confirmed using Ag-impregnation and FISH with an rDNA probe. In general, the NOR-bearing chromosomes corresponded to chromosome 8, but NORs were found on chromosome 3 or 4 in some species. Leptodactylus marmoratus had NORs on chromosome pairs 6 and 8. The data from C-banding, fluorochrome staining, and FISH using the telomeric probe helped in characterising the repetitive sequences. Even though hybridisation did occur on the chromosome ends, telomere-like repetitive sequences outside of the telomere region were identified. Metaphase I cells from L. pentadactylus confirmed its complex karyotype constitution because 12 chromosomes appeared as ring-shaped chain in addition to five bivalents.

Conclusions

Species of Leptodactylus exhibited both major and minor karyotypic differences which were identified by classical and molecular cytogenetic techniques. Replication banding, which is a unique procedure that has been used to obtain longitudinal multiple band patterns in amphibian chromosomes, allowed us to outline the general mechanisms responsible for these karyotype differences. The findings also suggested that L. marmoratus, which was formerly included in the genus Adenomera, may have undergone great chromosomal repatterning.

     

Effect of Vitamin E on Blood-Brain Barrier Permeability in Aged Rats with PTZ-Induced Convulsions

The effect of vitamin E on blood-brain barrier (BBB) permeability was studied under conditions of pentylenetetrazole (PTZ)-induced convulsions in aged (23- to 24-month-old) male albino rats; Evans Blue was used as a tracer. The BBB permeability was found to increase considerably in rats with PTZ-evoked seizures; the Evans Blue contents in the left and right hemispheres and cerebellum + brainstem region were significantly higher than those in the control. Vitamin E at a dose of 70 mg/kg exerted practically no beneficial effect on the increased BBB permeability in rats with seizures, while a greater dose of vitamin E (700 mg/kg) exerted a significant protective effect, especially with respect to the cerebellum + brainstem regions (P < 0.01). The seizure-related rise in the arterial blood pressure was also smaller in the latter experimental group. Thus, our observations confirm the importance of the vitamin E dose as a protective factor for BBB permeability and demonstrate that the dose dependence of this antioxidant in aged animals differs from that in younger organisms.     

An electrically-controlled programmable microfluidic concentration waveform generator

Background

Biological systems have complicated environmental conditions that vary both spatially and temporally. It becomes necessary to impose time-varying soluble factor concentrations to study such systems, including cellular responses to pharmaceuticals, inflammation with waxing and waning cytokine concentrations, as well as circadian rhythms and their metabolic manifestations. There is therefore a need for platforms that can achieve time-varying concentrations with arbitrary waveforms.

Results

To address this need, we developed a microfluidic system that can deliver concentration waveforms in a fast and accurate manner by adopting concepts and tools from electrical engineering and fluid mechanics. Specifically, we employed pulse width modulation (PWM), a commonly used method for generating analog signals from digital sources. We implement this technique using three microfluidic components via laser ablation prototyping: low-pass filter (lower frequency signals permitted, high frequency signals blocked), resistor, and mixer. Each microfluidic component was individually studied and iteratively tuned to generate desired concentration waveforms with high accuracy. Using fluorescein as a small-molecule soluble factor surrogate, we demonstrated a series of concentration waveforms, including square, sawtooth, sinusoidal, and triangle waves with frequencies ranging from 100 mHz to 400 mHz.

Conclusion

We reported the fabrication and characterization of microfluidic platform that can generate time-varying concentrations of fluorescein with arbitrary waveforms. We envision that this platform will enable a wide range of biological studies, where time-varying soluble factor concentrations play a critical role. In addition, the technology is expected to assist in the development of biomedical devices that allow precise dosing of pharmaceuticals for enhanced therapeutic efficacy and reduced toxicity.

     

Lifelong Consumption of Sodium Selenite: Gender Differences on Blood–Brain Barrier Permeability in Convulsive,Hypoglycemic Rats

The aim of this study was to compare the effects of hypoglycemia and induced convulsions on the blood-brain barrier permeability in rats with or without lifelong administration of sodium selenite. There is a significant decrease of the blood-brain barrier permeability in three brain regions of convulsive, hypoglycemic male rats treated with sodium selenite when compared to sex-matched untreated rats (p<0.05), but the decrease was not significant in female rats (p>0.05). The blood-brain barrier permeability of the left and right hemispheres of untreated, moderately hypoglycemic convulsive rats of both genders was better than their untreated counterparts (p<0.05). Our results suggest that moderate hypoglycemia and lifelong treatment with sodium selenite have a protective effect against blood-brain barrier permeability during convulsions and that the effects of sodium selenite are gender-dependent.     

Inhibition of Sulfate Respiration by 1,8-Dihydroxyanthraquinone and Other Anthraquinone Derivatives

Derivatives of 9,10-anthracenedione, or anthraquinone, were shown to inhibit respiratory sulfate reduction by pure cultures of sulfate-reducing bacteria, as well as by crude enrichment cultures. Structure-activity studies showed that an increasing degree of substitution of the anthraquinone nucleus resulted in increasing 50% inhibition (I(inf50)) values for sulfate respiration. Addition of charged ring substituents also resulted in an increase in the I(inf50) concentration. Experiments carried out with 1,8-dihydroxyanthraquinone demonstrated inhibition of hydrogen-dependent sulfate respiration but not hydrogen-dependent sulfite or thiosulfate respiration. Addition of pyruvate resulted in stimulation of sulfate-dependent hydrogen oxidation in the presence of the anthraquinone. These observations, together with a direct demonstration of uncoupling in French press vesicle preparations, suggest that the underlying mechanism of inhibition is uncoupling of ATP synthesis from electron transfer reactions. The low I(inf50) values for inhibition (0.5 to 10 (mu)M) and the relatively low general toxicity of anthraquinones suggest that these compounds may be useful for inhibition of sulfide generation in situations which are incompatible with the use of broadly toxic biocides.     

A novel knowledge-based approach to design inorganic-binding peptides

MOTIVATION: The discovery of solid-binding peptide sequences is accelerating along with their practical applications in biotechnology and materials sciences. A better understanding of the relationships between the peptide sequences and their binding affinities or specificities will enable further design of novel peptides with selected properties of interest both in engineering and medicine. RESULTS: A bioinformatics approach was developed to classify peptides selected by in vivo techniques according to their inorganic solid-binding properties. Our approach performs all-against-all comparisons of experimentally selected peptides with short amino acid sequences that were categorized for their binding affinity and scores the alignments using sequence similarity scoring matrices. We generated novel scoring matrices that optimize the similarities within the strong-binding peptide sequences and the differences between the strong- and weak-binding peptide sequences. Using the scoring matrices thus generated, a given peptide is classified based on the sequence similarity to a set of experimentally selected peptides. We demonstrate the new approach by classifying experimentally characterized quartz-binding peptides and computationally designing new sequences with specific affinities. Experimental verifications of binding of these computationally designed peptides confirm our predictions with high accuracy. We further show that our approach is a general one and can be used to design new sequences that bind to a given inorganic solid with predictable and enhanced affinity.