Adaptation of Plasmodium vivax to growth in owl monkeys (Aotus nancymai)

The purpose of this study was reactivation and adaptation of a strain of Plasmodium vivax to Aotus nancymai monkeys. A need arose for malarial parasites for use in serologic and molecular studies and for teaching slides. This particular strain of parasite had been characterized previously as producing high-density parasitemia in splenectomized New World monkeys and therefore represented a good candidate for reactivation. P. vivax (Vietnam II), isolated in 1970, was reactivated after adaptation in Aotus lemurinus griseimembra monkeys nearly 33 years earlier and adapted to A. nancymai monkeys. Passage was achieved by intravenous inoculation of parasite blood stages into splenectomized A. nancymai monkeys. Parasitemia was determined by analyzing daily blood smears stained with Giemsa. Maximum parasite counts ranged from 10,630 to 94,000 parasites/microl; the mean maximum parasite count for the four animals was 39,565 parasites/microl. Parasite counts of > 10,000/microl were maintained for 2 to 64 days. After only three passages of the parasite, attempts to reactive were successful. A. nancymai proved a suitable animal model for the recovery of this parasite. In conclusion, successful reactivation and adaptation of this parasite offers the capability to perform a series of diagnostic, immunologic, and molecular studies as well as to provide otherwise potentially unavailable teaching materials to healthcare professionals.     

High‐Capacity All‐Solid‐State Sodium Metal Battery with Hybrid Polymer Electrolytes

All‐solid‐state sodium metal batteries (SSMBs) are of great interest for their high theoretical capacity, nonflammability, and relatively low cost owing partially to the abundance of sodium recourses. However, it is challenging to fabricate SSMBs because compared with their counterparts, which contain lithium metal, sodium metal is mechanically softer and more reactive toward the electrolyte. Herein, the synthesis and electrochemical properties of newly designed sodium‐containing hybrid network solid polymer electrolytes (SPEs) and their application in SSMBs are reported. The hybrid network is synthesized by controlled crosslinking of octakis(3‐glycidyloxypropyldimethylsiloxy)octasilsesquioxane and amine‐terminated polyethylene glycol in existence with sodium perchlorate (NaClO₄). Plating and stripping experiments using symmetric cells show prolonged cycle life of the SPEs, >5150 and 3550 h at current density of 0.1 and 0.5 mA cm^?2, respectively. The results for the first time show that the SPE|sodium metal interface migrates into the SPE phase upon cycling. SSMBs fabricated with the hybrid SPE sandwiched between sodium metal anode and bilayered δ‐Na_xV₂O₅ cathode exhibit record‐high specific capacity for solid sodium‐ion batteries of 305 mAh g^?1 and excellent Coulombic efficiency. This work demonstrates that the hybrid network SPEs are promising for SSMB applications.     

Stress and developmental regulation of the yeast C-type cyclin Ume3p (Srb11p/Ssn8p).

The ume3-1 allele was identified as a mutation that allowed the aberrant expression of several meiotic genes (e.g. SPO11, SPO13) during mitotic cell division in Saccharomyces cerevisiae. Here we report that UME3 is also required for the full repression of the HSP70 family member SSA1. UME3 encodes a non-essential C-type cyclin (Ume3p) whose levels do not vary through the mitotic cell cycle. However, Ume3p is destroyed during meiosis or when cultures are subjected to heat shock. Ume3p mutants resistant to degradation resulted in a 2-fold reduction in SPO13 mRNA levels during meiosis, indicating that the down-regulation of this cyclin is important for normal meiotic gene expression. Mutational analysis identified two regions (PEST-rich and RXXL) that mediate Ume3p degradation. A third destruction signal lies within the highly conserved cyclin box, a region that mediates cyclin-cyclin-dependent kinase (Cdk) interactions. However, the Cdk activated by Ume3p (Ume5p) is not required for the rapid destruction of this cyclin. Finally, Ume3p destruction was not affected in mutants defective for ubiquitin-dependent proteolysis. These results support a model in which Ume3p, when exposed to heat shock or sporulation conditions, is targeted for destruction to allow the expression of genes necessary for the cell to respond correctly to these environmental cues.     

Bioavailability of selenium from veal, chicken, beef, pork, lamb, flounder, tuna, selenomethionine, and sodium selenite assessed in selenium-deficient rats

The bioavailability of selenium (Se) from veal, chicken, beef, pork, lamb, flounder, tuna, selenomethionine (SeMet), and sodium selenite was assessed in Se-deficient Fischer-344 rats. Se as veal, chicken, beef, pork, lamb, flounder, tuna, SeMet, and sodium selenite was added to torula yeast (TY) basal diets to comprise Se-inadequate (0.05 mg Se/kg) diets. Se as sodium selenite was added to a TY basal diet to comprise a Se-adequate (0.10 mg Se/kg), Se-control diet. The experimental diets were fed to weanling Fischer-344 rats that had been subjected to dietary Se depletion for 6 wk. After 9 wk of the dietary Se repletion, relative activity of liver glutathione peroxidase (GSHPx) from the different dietary groups compared with control rats (100%) was: flounder 106%, tuna 101%, pork 86%, sodium selenite 81%, SeMet 80%, beef 80%, chicken 77%, veal 77%, and lamb 58%. Se from flounder was the most efficient at restoring Se concentrations in the liver and skeletal muscle. Se from sodium selenite, SeMet, beef, veal, chicken, pork, lamb, and tuna was not dietarily sufficient to restore liver and muscle Se after 9 wk of recovery following a 6-wk period of Se depletion.     

Mutational analysis of the role of glycoprotein I in varicella-zoster virus replication and its effects on glycoprotein E conformation and trafficking.

The contributions of the glycoproteins gI (ORF67) and gE (ORF68) to varicella-zoster virus (VZV) replication were investigated in deletion mutants made by using cosmids with VZV DNA derived from the Oka strain. Deletion of both gI and gE prevented virus replication. Complete deletion of gI or deletions of 60% of the N terminus or 40% of the C terminus of gI resulted in a small plaque phenotype as well as reduced yields of infectious virus. Melanoma cells infected with gI deletion mutants formed abnormal polykaryocytes with a disrupted organization of nuclei. In the absence of intact gI, gE became localized in patches on the cell membrane, as demonstrated by confocal microscopy. A truncated N-terminal form of gI was transported to the cell surface, but its expression did not restore plaque morphology or infectivity. The fusogenic function of gH did not compensate for gI deletion or the associated disruption of the gE-gI complex. These experiments demonstrated that gI was dispensable for VZV replication in vitro, whereas gE appeared to be required. Although VZV gI was dispensable, its deletion or mutation resulted in a significant decrease in infectious virus yields, disrupted syncytium formation, and altered the conformation and distribution of gE in infected cells. Normal cell-to-cell spread and replication kinetics were restored when gI was expressed from a nonnative locus in the VZV genome. The expression of intact gI, the ORF67 gene product, is required for efficient membrane fusion during VZV replication.     

Metabolism of 14C-Chlorobenzilate and 14C-Chloropropylate by Rhodotorula gracilis

Rhodotorula gracilis metabolizes Chlorobenzilate (ethyl 4,4'-dichlorobenzilate) and Chloropropylate (isopropyl 4,4'-dichlorobenzilate) to several metabolites in a basal medium supplemented by sucrose and by several intermediates of the citric acid cycle. Three identified metabolites resulting from the degradation of either acaricide, were 4,4'-dichlorobenzilic acid, 4,4'-dichlorobenzophenone, and carbon dioxide. Chlorobenzilate, i.e., ethyl ester of 4,4'-dichlorobenzilic acid, was more easily hydrolyzed than Chloropropylate, i.e., isopropyl ester of this acid, so that larger amounts of carbon dioxide and 4,4'-dichlorobenzophenone were obtained from Chlorobenzilate degradation. Regardless of acaricides used, longer incubation caused a higher accumulation of 4,4'-dichlorobenzophenone. The probable steps of the degradation pathway are: Chlorobenzilate (or Chloropropylate) --> 4,4'-dichlorobenzilic acid --> 4,4'-dichlorobenzophenone plus carbon dioxide. It appears that the decarboxylation of 4,4'-dichlorobenzilic acid to 4,4'-dichlorobenzophenone was hindered by alpha-ketoglutarate and enhanced by succinate.     

Aggressive neighbors and dense nesting: nest site choice and success in high-Arctic common eiders

Minimizing the risk of nest predation has led some bird species to exploit the nest defense behavior of other species. At Nasaruvaalik Island, Nunavut, Canada, some common eiders (Somateria mollissima borealis) nest within the boundaries of Arctic tern (Sterna paradisaea) colonies, while others nest elsewhere on the island, away from the terns. We tested the effects of location (within vs. outside the tern colonies), density of common eider nests, and annual variation on the nesting parameters of common eiders. Our results suggest that nesting in association with Arctic terns does not confer an obvious benefit to eiders. Such associative nesting of eiders and terns may be the result of overlapping habitat preferences between the two species, or a general scarcity of suitable nesting habitat for ground-nesting species in the high Arctic. However, eiders nesting in higher densities with other eiders had greater nest success and lower total clutch predation, indicating a positive correlation between nest density and success.     

Expression and characterization of human apolipoprotein A-I in Chinese hamster ovary cells

We produced human apolipoprotein A-I (apoA-I) in Chinese hamster ovary (CHO) cells. The CHO cells were transfected with an expression plasmid which placed the human apoA-I gene under the direction of the human metallothionein II gene promoter. Isolation of a clonal cell line resulted in high level expression of apoA-I. Greater than 30% of total protein secreted by these CHO cells was apoA-I, which enabled us to purify apoA-I with a single step purification scheme. As a result, large quantities of apoA-I can be produced and isolated without having to rely on plasma sources. Structural characterization of the recombinant apoA-I showed it to be identical to authentic apoA-I from human serum high density lipoprotein. Furthermore, we demonstrated approximately equal to 90% of the apoA-I secreted by CHO cells is processed, mature protein. A portion of the secreted recombinant apoA-I was associated with lipid and floated at a density approximately equal to 1.10 g/ml. Additional analysis identified the presence of five isoforms of apoA-I in the CHO cell conditioned medium. Processing and post-translational modification of the recombinant apoA-I occurred in the CHO cell cultures in the absence of serum components. We conclude that the human apoA-I produced by CHO cells is identical to circulating, mature apoA-I in humans and that recombinant mammalian expression offers an opportunity to investigate apoA-I processing.     

Timing of Environmental Enrichment Affects Memory in the House Cricket,Acheta domesticus

Learning appears to be ubiquitous among animals, as it plays a key role in many behaviors including foraging and reproduction. Although there is some genetic basis for differences in learning ability and memory retention, environment also plays an important role, as it does for any other trait. For example, adult animals maintained in enriched housing conditions learn faster and remember tasks for longer than animals maintained in impoverished conditions. Such plasticity in adult learning ability has often been linked to plasticity in the brain, and studies aimed at understanding the mechanisms, stimuli, and consequences of adult behavioral and brain plasticity are numerous. However, the role of experiences during post-embryonic development in shaping plasticity in adult learning ability and memory retention remain relatively unexplored. Using the house cricket (Acheta domesticus) as a model organism, we developed a protocol to allow the odor preference of a large number of crickets to be tested in a short period of time. We then used this new protocol to examine how enrichment or impoverishment at two developmental stages (either the last nymphal instar or young adult) affected adult memory. Our results show that regardless of nymphal rearing conditions, crickets that experienced an enriched rearing condition as young adults performed better on a memory task than individuals that experienced an impoverished condition. Older adult crickets (more than 1 week post adult molt) did not demonstrate differences in memory of the odor task, regardless of rearing condition as a young adult. Our results suggest that environmentally-induced plasticity in memory may be restricted to the young adult stage.     

Advances in the Conceptualization and Measurement of Health Care Empowerment: Development and Validation of the Health Care Empowerment Inventory

The Health Care Empowerment Model offers direction for the investigation of patient-controlled engagement and involvement in health care. At the core of the model is the construct of Health Care Empowerment (HCE), for which there exist no validated measures. A set of 27 candidate self-report survey items was constructed to capture five hypothesized inter-related facets of HCE (informed, engaged, committed, collaborative, and tolerant of uncertainty). The full item set was administered to 644 HIV-infected persons enrolled in three ongoing research studies. Exploratory and confirmatory factor analyses resulted in a two factor solution comprising four items each on two subscales: (1) HCE: Informed, Committed, Collaborative, and Engaged HCE ICCE) and (2) HCE Tolerance of Uncertainty (HCE TU). Subscale scores were evaluated for relationships with relevant constructs measured in the three studies, including depression, provider relationships, medication adherence, and HIV-1 viral load. Findings suggest the utility of this 8-item Health Care Empowerment Inventory (HCEI) in efforts to measure, understand, and track changes in the ways in which individuals engage in health care.