Observations onApoica pallens,a nocturnal neotropical social wasp (Hymenoptera: Vespidae,Polistinae, Epiponini)

Summary Colonies and nests ofApoica pallens in the llanos region of Venezuela range from small foundress nests to large mature colonies. Nests are sited on small diameter, near-horizontal branches in a variety of shrub and tree species. During the day, adult wasps cluster on the face of the nest in an array that seems to be determined by orientation to gravity; defense of the colony against parasitoids and ants by the resting wasps may be more a passive than an active behavior. Wasps fan their wings to cool the colony during the day, but no foraging for water accompanies the fanning behavior. Nightly foraging activity begins with the explosive departure from the nest of hundreds of wasps, most of which rapidly return. Moderate foraging levels early at night give way to very low foraging levels in pre-dawn hours. The period of moderate foraging may be extended for longer hours during increased moonlight. Foraging wasps collect arthropod provisions for larvae. Larvae produce a trophallactic saliva; adults engage in inter-adult trophallaxis; brood are cannibalized. During cluster formation prior to swarm emigration, adult wasps do not appear to scent-mark substrates such as leaves. Instead,A. pallens exhibits a calling behavior, unique among polistine wasps studied to date, in which the gaster is held rigidly away from the thorax and metasomal sternal glands are exposed. Swarms can emigrate during the day.A. pallens may incorporate absconding and colony relocation as features of its colony cycle in the highly seasonal llanos.     

The effect of cycloheximide on Euglena gracilis phenylalanyl-tRNA synthetases

The effect of cycloheximide on the chloroplastic, cytoplasmic and mitochondrial phenylalanyltransferRNA synthetases of Euglena gracilis was studied by growing both logarithmic and stationary phase cultures in the presence of the antibiotic. Enzyme activity was measured relative to untreated control cultures. At very low concentrations of cycloheximide (1 g/ml), all three log phase enzymes showed an increase in activity of 40–50%. At slightly higher concentrations (2.5 g/ml), the phenylalanyl-tRNA synthetase activities were comparable to those of the control cultures. At a cycloheximide concentration of 5g/ml the enzyme activities from stationary phase cultures showed only very slight decreases (5–20%). The cytoplasmic and mitochondrial enzymes behaved similarly in log phase cultures at this concentration. However, the chloroplastic phenylalanyl-tRNA synthetase from log phase cultures treated with 5g/ml cycloheximide showed a marked decrease in activity (70%). A further increase in antibiotic concentration to 10g/ml resulted in significant losses of activity of all three enzymes, from both growth stages. The implications of the data with regard to identification of the site(s) of chloroplast enzyme synthesis are discussed.     

Clavispora,a new yeast genus of the Saccharomycetales

In Candida lusitaniae van Uden et do Carmo-Sousa (1959), strains of opposite sex have been found. Cells of the opposite mating types conjugate and form asci with one to four clavate spores. These are easily liberated from the ascus. The type strain of Candida obtusa (Dietrichson) van Uden et do Carmo-Sousa ex van Uden et Buckley (1970) also produces ascospores after mating with one of the strains of Candida lusitaniae. As clavate ascospores are unknow in yeasts, a new genus, Clavispora, is proposed with Clavispora lusitaniae spec. nov. as type species.     

Determination of the chromosomal size of three different strains of Enterococcus faecalis and one strain of Enterococcus faecium.

Pulsed-field gel electrophoresis was used to determine the chromosomal size of three different strains of Enterococcus faecalis and one strain of Enterococcus faecium. The size determinations of OG1X, a strain of E. faecalis widely used in many laboratories for genetic studies, using Sma I, Not I, and Sfi I alone or in combination, ranged from 2,750 to 2,761 kb. Using the same enzymes as with OG1X, the size of HH-67, a plasmid-free clinical isolate of E. faecalis, was determined to be 2,170-2,288 kb and the size of JH2-2, an E. faecalis recipient strain, ranged from 2,008 to 2,135 kb. The size range generated for GE-1, a plasmid-free E. faecium strain, with the use of Sma I, Not I, and Apa I was 2,045-2,155 kb. Although OG1X differed in size from the other three enterococci, each individual enterococcal strain generated reproducible results in different experiments. However, for both E. faecalis OG1X and E. faecium GE-1, one of the enzymes used generated a considerably smaller molecular size than that generated by the other two enzymes. The discrepancy was due to visually undiscernible comigrating fragments, and serves to point out a potential source of error if fewer than two enzymes are used to size a genome. The size discrepancies were resolved by digesting individual fragments with a second enzyme. The molecular sizes of these enterococcal strains are larger than that recently reported for Campylobacter, smaller than that of Escherichia coli and Pseudomonas aeruginosa, and similar (OG1X) or smaller (JH2-2, HH67, and GE-1) than the 2,819-kb reported for Streptococcus mutans.     

Effect of gestational and postnatal environmental temperature on metabolic rate in the altricial rodent, Phyllotis darwini

1. In the altricial rodent, Phyllotis darwini, we found higher body temperatures and faster developmental rates of the thermoregulatory capacity in neonates born from cold- than warm-acclimated mothers.

2. This difference could be explained by maternal effects on the litter, such as high levels of catecholamines and thyroxin levels, high concentration of the uncoupled protein and larger quantity of brown adipose tissue as a consequence of cold acclimation.

3. The exposition of mothers and the maintenance of cold condition during the early development might be responsible of the high metabolism and better thermoregulatory capacity of newborns.

Electrocoagulation as a clarifiation and pre-purification step in plant based bioprocesses

Electrocoagulation is a technique that has been applied to water and wastewater treatment. We propose here extension of this technique to the field of plant downstream processing (DSP), where plants are used as a bioreactors for recombinant protein production. The main problem in plant-based bioreactors is the presence of chlorophyll and phenolic compounds in plant extracts, which tend to precipitate and denature proteins. Their removal from the extracts is essential, since they have a large impact on the DSP performance. In the present work we studied the application of an electrocoagulation based technique to clarify tobacco leaf extracts by removing chlorophyll and phenolic compounds. By manipulating the pH of electrocoagulation, 90% of chlorophyll, 65% of phenolic compounds, and only 23% of total protein were removed from the extract. The process preferentially removes proteins with more acidic pI (below 6.03) and the pH of the process differentially effects removal of acidic and basic proteins.     

FK506 binding protein 8 peptidylprolyl isomerase activity manages a late stage of cystic fibrosis transmembrane conductance regulator (CFTR) folding and stability

Cystic fibrosis (CF) is caused by mutations in the apical chloride channel cystic fibrosis transmembrane conductance regulator (CFTR) with 90% of patients carrying at least one deletion of the F508 (ΔF508) allele. This mutant form of CFTR is characterized by a folding and trafficking defect that prevents exit from the endoplasmic reticulum. We previously reported that ΔF508 CFTR can be recovered in a complex with Hsp90 and its co-chaperones as an on-pathway folding intermediate, suggesting that Δ508 CF disease arises due to a failure of the proteostasis network (PN), which manages protein folding and degradation in the cell. We have now examined the role of FK506-binding protein 8 (FKBP8), a component of the CFTR interactome, during the biogenesis of wild-type and ΔF508 CFTR. FKBP8 is a member of the peptidylprolyl isomerase family that mediates the cis/trans interconversion of peptidyl prolyl bonds. Our results suggest that FKBP8 is a key PN factor required at a post-Hsp90 step in CFTR biogenesis. In addition, changes in its expression level or alteration of its activity by a peptidylprolyl isomerase inhibitor alter CFTR stability and transport. We propose that CF is caused by the sequential failure of the prevailing PN pathway to stabilize ΔF508-CFTR for endoplasmic reticulum export, a pathway that can be therapeutically managed.     

Antigen Recognition By Autoreactive Cd4+ Thymocytes Drives Homeostasis Of The Thymic Medulla

The thymic medulla is dedicated for purging the T-cell receptor (TCR) repertoire of self-reactive specificities. Medullary thymic epithelial cells (mTECs) play a pivotal role in this process because they express numerous peripheral tissue-restricted self-antigens. Although it is well known that medulla formation depends on the development of single-positive (SP) thymocytes, the mechanisms underlying this requirement are incompletely understood. We demonstrate here that conventional SP CD4⁺ thymocytes bearing autoreactive TCRs drive a homeostatic process that fine-tunes medullary plasticity in adult mice by governing the expansion and patterning of the medulla. This process exhibits strict dependence on TCR-reactivity with self-antigens expressed by mTECs, as well as engagement of the CD28-CD80/CD86 costimulatory axis. These interactions induce the expression of lymphotoxin α in autoreactive CD4⁺ thymocytes and RANK in mTECs. Lymphotoxin in turn drives mTEC development in synergy with RANKL and CD40L. Our results show that Ag-dependent interactions between autoreactive CD4⁺ thymocytes and mTECs fine-tune homeostasis of the medulla by completing the signaling axes implicated in mTEC expansion and medullary organization.