Glycosome assembly in trypanosomes: variations in the acceptable degeneracy of a COOH-terminal microbody targeting signal

Trypanosomes compartmentalize most of their glycolytic enzymes in a peroxisome-like microbody, the glycosome. The specificity of glycosomal targeting was examined by expression of chloramphenicol acetyltransferase fusion proteins in trypanosomes and monkey cells. Compartmentalization was assessed by cell fractionation, differential detergent permeabilization, and immunofluorescence. The targeting signal of trypanosome phosphoglycerate kinase resides in the COOH-terminal hexapeptide, NRWSSL; a basic amino acid is not required. The minimal targeting signal is, as for mammalian cells, a COOH-terminal tripeptide related to -SKL. However, the acceptable degeneracy of the signal for glycosomal targeting in trypanosomes is considerably greater than that for peroxisomal targeting in mammals, with particularly relaxed requirements in the penultimate position.     

Toward a clinical antifungal peptoid: Investigations into the therapeutic potential of AEC5

Cryptococcus neoformans is a fungal pathogen that causes cryptococcal meningitis in immunocompromised individuals. Existing antifungal treatment plans have high mammalian toxicity and increasing drug resistance, demonstrating the dire need for new, nontoxic therapeutics. Antimicrobial peptoids are one alternative to combat this issue. Our lab has recently identified a tripeptoid, AEC5, with promising efficacy and selectivity against C. neoformans. Here, we report studies into the broad-spectrum efficacy, killing kinetics, mechanism of action, in vivo half-life, and subchronic toxicity of this compound. Most notably, these studies have demonstrated that AEC5 rapidly reduces fungal burden, killing all viable fungi within 3 hours. Additionally, AEC5 has an in vivo half-life of 20+ hours and no observable in vivo toxicity following 28 days of daily injections. This research represents an important step in the characterization of AEC5 as a practical treatment option against C. neoformans infections.     

Screening of marine actinomycetes isolated from the Bay of Bengal,India for antimicrobial activity and industrial enzymes

A total of 288 marine samples were collected from different locations of the Bay of Bengal starting from Pulicat lake to Kanyakumari, and 208 isolates of marine actinomycetes were isolated using starch casein agar medium. The growth pattern, mycelial coloration, production of exopolysaccharides and diffusible pigment and abundance of Streptomyces spp. were documented. Among marine actinomycetes, Streptomyces spp. were present in large proportion (88%). Among 208 marine actinomycetes, 111 isolates exhibited antimicrobial activity against human pathogens, and 151 showed antifungal activity against two plant pathogens. Among 208 isolates, 183, 157, 116, 72 and 68 isolates produced lipase, caseinase, gelatinase, cellulase and amylase, respectively. The results of diversity, antimicrobial activity and enzymes production have increased the scope of finding industrially important marine actinomycetes from the Bay of Bengal and these organisms could be vital sources for the discovery of industrially useful molecules/enzymes.     

Plant regeneration from sorghum anther cultures and field evaluation of progeny

Summary Embryogenic calli were obtained from a hybrid line of Sorghum bicolor (L.) Moench anthers with mid to late uninucleate pollen cultured in N₆ medium supplemented with 3.0% sucrose, 2.0 mg/1 2,4-D, 0.8% agar and incubated at 30°C, which was the optimum temperature. The regeneration of embryos was obtained from the embryogenic calli cultured in modified MS medium supplemented with 3.0% sucrose, 2.0 mg/1 BAP combined with 0.3 mg/1 IAA and 0.8% agar. A total of 248 doubled haploids and 12 haploid plants were regenerated. In a subsequent field study, the selfed progeny from anther culture (designated as the anther culture-2, [A₂] generation) derived families was compared with both the F₂ and the F₁ for agronomic and morphological traits. Significant differences were noticed between the family means of both A₂ and F₂ for all the quantitative traits studied. The distinctive difference in the behavior of the A₂ families in comparison with the F₂ was established by within family variance, which was significant in F₂ for days to 50 per cent flowering, plant height, panicle length, leaf area index, dry matter production, harvest index and grain yield and was non-significant in A₂. Male sterility, one of the potentially important traits, currently exploited in the hybrid seed production of cereals, including CSH5 hybrid sorghum and the morphological traits (panicle shape, compactness, grain color, glume color and nature of the leaf sheath) segregated in the F₂. Such segregation was not observed within A₂ families and they bred true to their respective A₁ plants, indicating the rapid attainment of homozygosity/uniformity. The present study establishes the gametophytic origin of anther culture derived families and indicates the possibility of rapid production of homozygous lines which can be used as recombinant inbreds.Abbreviations N₆ Chu et al. (1975) - MS Murashige and Skoog (1962) - 2,4-D, 2 4-Dichlorophenoxyacetic acid - IAA Indole-3 -acetic acid - BAP Benzylaminopurine - A₁ regenerated plants - A2 selfed progeny from A₁     

A comparison of mutation induction in diploid and tetraploid rice

Summary A comparative assessment of the frequency and spectrum of chlorophyll mutations in the M ₂ and M ₃ of three diploids and one tetraploid of rice after X-irradiation was made. As well as a linear relationship of the frequency of mutations with the dose of mutagen, a saturation effect was also evident. Among the three diploids, the maximum frequency of mutations was observed in T. N. 1, followed by G. E. B. 24 and A. S. D. 8. The diploid of G. E. B. 24 showed a higher frequency of mutations than the tetraploid when measured on the M₁ plant basis only. The diploids showed a higher rate of mutations in the M ₂ than in the M₃.There was no relationship between the frequency of different types of chlorophyll mutations and the dose of X-rays. Albina, occurred in greater proportion than the other types of mutations in the diploids as well as in the tetraploid. The diploid showed a wider spectrum in the M ₂, whereas the tetraploid recorded the maximum types of mutations in the M ₃. Striking differences in the spectrum and the relative frequency of each type were observed among the three diploids and also between the diploid G. E. B. 24 and its tetraploid.The frequency and spectrum of induced mutations in the diploid and autotetraploid provided an insight into the genetic behaviour of the diploid and autotetraploid of G. E. B. 24, indicating that the genetics of diploidisation of the existing diploid rice may give evidence on the nature of the evolutionary pathway.