Comparative histopathology of three ascovirus isolates in larval noctuids

The histopathology caused by three ascoviruses isolated respectively from Heliothis zea, Spodoptera frugiperda, and Trichoplusia ni was studied in the host species from which the viruses were isolated originally, or in the case of the H. zea isolate, in H. virescens. In all three isolates, infected cells and virion-containing vesicles in the hemolymph were observed by 3 days postinoculation. The isolates from H. zea and T. ni exhibited relatively broad tissue tropisms infecting the tracheal matrix, epidermis, and connective tissue, and in the latter isolate, the fat body. Although considered variants of the same virus, these isolates varied slightly in their tissue tropism in that the isolate from H. zea replicated more extensively in the epidermis. The isolate from T. ni replicated in the fat body during early stages of disease, but not in advanced stages, and was not as common in the epidermis as the isolate from H. zea. The isolate from S. frugiperda replicated only in the fat body and completely destroyed this tissue by 12 days postinoculation. The progression of disease within the tissues was correlated with a rise in the concentration of virion-containing vesicles in the hemolymph of all three isolates. Vesicle concentrations rose from 10(7) on day 2 to 10(8)/ml of hemolymph on day 9, plateaued for 5-7 days, and then declined gradually through day 20. These results indicate that the chronic nature of the diseases caused by ascoviruses is due to the limited degree to which they infect major tissues (isolates from T. ni and H. zea) or the infection of tissues less essential to the maintenance of larval life (isolate from S. frugiperda).     

Surface Acidity and Solid-State Compatibility of Excipients with an Acid-Sensitive API: Case Study of Atorvastatin Calcium

The objectives of this study were to measure the apparent surface acidity of common excipients and to correlate the acidity with the chemical stability of an acid-sensitive active pharmaceutical ingredient (API) in binary API-excipient powder mixtures. The acidity of 26 solid excipients was determined by two methods, (i) by measuring the pH of their suspensions or solutions and (ii) the pH equivalent (pH_eq) measured via ionization of probe molecules deposited on the surface of the excipients. The chemical stability of an API, atorvastatin calcium (AC), in mixtures with the excipients was evaluated by monitoring the appearance of an acid-induced degradant, atorvastatin lactone, under accelerated storage conditions. The extent of lactone formation in AC-excipient mixtures was presented as a function of either solution/suspension pH or pH_eq. No lactone formation was observed in mixtures with excipients having pH_eq > 6, while the lactone levels were pronounced (> 0.6% after 6 weeks at 50°C/20% RH) with excipients exhibiting pH_eq < 3. The three pH_eq regions (> 6, 3–6, and < 3) were consistent with the reported solution pH-stability profile of AC. In contrast to the pH_eq scale, lactone formation did not show any clear trend when plotted as a function of the suspension/solution pH. Two mechanisms to explain the discrepancy between the suspension/solution pH and the chemical stability data were discussed. Acidic excipients, which are expected to be incompatible with an acid-sensitive API, were identified based on pH_eq measurements. The incompatibility prediction was confirmed in the chemical stability tests using AC as an example of an acid-sensitive API.

Electronic supplementary material

The online version of this article (doi:10.1208/s12249-014-0231-7) contains supplementary material, which is available to authorized users.KEY WORDS: acidity, atorvastatin, excipients, pH indicators, solid-state stability     

Ectoparasite Raymondia lobulata infestation in relation to the reproductive cycle of its host--the greater false vampire bat Megaderma lyra

To study variation of infestations by the bat fly Raymondia lobulata (Diptera: Streblidae) on the greater false vampire bat Megaderma lyra (Chiroptera: Megadermatidae), we captured individual bats at their day roost in the south of India and recorded their rate of infestation continuously for a year. All examined bats (n = 72 individuals, 202 captures) were infested with parasites (n = 3,008). However, the recorded intensity of infestation (range 1-33) was gender-related and statistically higher in females than in males (F(1, 200) = 304.45, P < 0.001). Furthermore, pregnant and lactating females had greater parasite loads than non-reproductive females and males (F(1, 63) = 23.34, P < 0.001 and F(1, 37) = 78.07, P < 0.001, respectively). No significant differences were observed between males either during mating and non-mating periods or breeding and non-breeding seasons. Analysis of the relationship between parasite infestation and the reproductive status of bats revealed that pregnant and lactating females with pups were more vulnerable hosts for parasites. Our results also suggest a well-developed coevolutionary strategy for synchronized reproduction within the host-parasite relationship and add to our understanding of how host sex and reproductive status shape the dynamics of parasitism.     

Efficient synthesis and biological evaluation of proximicins A, B and C

A quick and efficient synthesis and the biological evaluation of promising antitumor-antibiotics proximicins A, B and C are reported. The characteristic repetitive unit of these molecules, the methyl 4-Boc-aminofuran-2-carboxylate 15, was prepared in three synthetic steps in good yield using an optimised copper-catalysed amidation method. The proximicins were evaluated for their antitumor activity using cellular methods. Proximicin B induced apoptosis in both Hodgkin's lymphoma and T-cell leukemia cell lines and proximicin C exhibited significantly high cytotoxicity against glioblastoma and breast carcinoma cells. The proximicins were also screened against Escherichia coli, Enterococcus faecalis and several strains of methicillin-and multidrug-resistant Staphylococcus aureus. Proximicin B showed noteworthy activity against antibiotic-resistant Gram-positive cocci.     

Environmental enrichment exerts anxiolytic effects in the Indian field mouse (Mus booduga)

Environmental enrichment (EE) is known to have behavioral and physiological anxiolytic effects in several animal models. However, it is as yet unclear how EE modulates behavior of wild animals and the underlying molecular mechanisms. The adult male field mouse Mus booduga (n = 42) captured at agricultural field, were housed in non-enriched standard condition (SC) for 7 days and considered as directly from wild (DW). Another two groups of mice were housed in either EE or SC for 30 days. Behavioral testing was carried out to assess their anxiety-like behavior in the elevated plus-maze (EPM). We found that on EPM, mice housed in EE display less anxiety like behavior when compared to mice housed in SC. Exposure to plus-maze did not increase the levels of corticosterone (CORT) in prefrontal cortex (PFC) and circulating CORT, and adrenocorticotropic hormone (ACTH) in the mice housed in EE but not in the mice housed in SC. We observed a trend in the EE induced inhibition of expression of corticotropin releasing hormone (CRH), glucocorticoid receptor (GR), E2 ubiquitin-conjugating enzyme (Ubc9) and steroid receptor coactivator-1 (SRC-1) mRNA levels, which are all known to be involved in the stress response signaling pathway. Our study suggests that EE exerts therapeutic and anxiolytic effects against stressors.     

N-fertilizer saving by the inoculation of Gluconacetobacter diazotrophicus and Herbaspirillum sp. in micropropagated sugarcane plants

Colonization of micropropagated sugarcane plants by Gluconacetobacter diazotrophicus and Herbaspirillum sp. was confirmed by a dot-immunoblot assay. In all, a 45-day short-term and 180-day long-term experiments conducted on micropropagated sugarcane plants of Co 86032, a sugar rich popular variety in South India, indicated the usefulness of these diazotrophs as plant growth promoting bacteria. Co-inoculation of these two bacteria enhanced the biomass considerably under N-limited condition in the short duration experiment. In the long-term experiment, the establishment of inoculated Herbaspirillum sp. remained stable with the age of the crop up to 180 days, while there was a reduction in population of G. diazotrophicus for the same period. The total bio-mass and leaf N were higher in plants inoculated with G. diazotrophicus and Herbaspirillum sp. without N fertilization and also in plants with 50% of the recommended N (140 kg ha(-1)) than the plants fertilized with recommended dose of inorganic N (280 kg ha(-1)). This experiment showed that inoculation with these bacteria in sugarcane variety Co 86032 could mitigate fertilizer N application considerably in sugarcane cultivation.     

Purification and characterization of a β-1,4-glucosidase from a newly isolated strain of <Emphasis Type="Italic">Fomitopsis pinicola</Emphasis>

An efficient ß-1,4-glucosidase (BGL) producing strain, Fomitopsis pinicola KMJ812, was isolated and identified based on morphological features and sequence analysis of internal transcribed spacer rDNA. An extracellular BGL was purified to homogeneity by sequential chromatography of F. pinicola culture supernatants on a DEAE-sepharose column, a gel filtration column, and then on a Mono Q column with fast protein liquid chromatography. The relative molecular weight of F. pinicola BGL was determined to be 105 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis, or 110 kDa by size exclusion chromatography, indicating that the enzyme is a monomer. The hydrolytic activity of the BGL had a pH optimum of 4.5 and a temperature optimum of 50°C. The enzyme showed high substrate specificity and high catalytic efficiency (k _cat?=?2,990 s^?1, K _m?=?1.76 mM, k _cat/K _m?=?1,700 mM^?1 s^?1) for p-nitrophenyl-β-d-glucopyranoside. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase family 3, indicating that the F. pinicola BGL is a member of glycoside hydrolase family 3. Although BGLs have been purified and characterized from several other sources, F. pinicola BGL is distinguished from other BGLs by its high catalytic efficiency and strict substrate specificity.