A new species of Eimeria Schneider, 1885 (Apicomplexa: Eimeriidae) from the eastern gray squirrel,Sciurus carolinensis (Rodentia: Sciuridae: Sciurinae: Sciurini) from Oklahoma,USA

Systematic Parasitology - A new species of Eimeria Schneider, 1885 is described from an eastern gray squirrel (Sciurus carolinensis) from McCurtain County, Oklahoma, USA. Oöcysts of Eimeria...     

Genetic effect of two APOA repeat polymorphisms (kringle 4 and pentanucleotide repeats) on plasma Lp(a) levels in American Samoans

Elevated plasma lipoprotein(a) [Lp(a)] level has been established as an independent risk factor for atherosclerosis and coronary heart disease. Considerable ethnic group differences in the distribution of plasma Lp(a) levels have raised public health concerns. Recently, we have reported that Samoans have the lowest plasma Lp(a) levels of any population group. In the present investigation, we report the contribution of two apolipoprotein(a) (APOA) polymorphisms, the kringle 4 type 2 (K4) repeat and the pentanucleotide repeat (PNR), in affecting plasma Lp(a) levels in an American Samoan sample (n = 309). The K4 repeats ranged in size from 15 to 40. The common alleles contained repeats ranging from 26 to 36 with allele frequencies between 5.5% to 9.7%, and these accounted for 82% of all alleles. An inverse relationship between K4 repeat number and plasma Lp(a) level was observed for single-banded (r = -0.59, p = 0.0001) and double-banded phenotypes (r = -0.50, p = 0.0001). This polymorphism explained 60% of the variation in plasma Lp(a) level in American Samoans. For the PNR polymorphism, five different repeat alleles and eight different genotypes were identified; the most common allele was eight repeats. The *8 PNR allele was associated with a wide range of K4 repeats, the *9 PNR allele with larger K4 repeats (25-40), and the *10 PNR with smaller K4 repeats (15-24). Analysis of variance (ANOVA) revealed that the PNR polymorphism accounts for 2.1% of the variability in plasma Lp(a) levels in this sample, when the K4 repeat polymorphism was taken into account. Our data show that common polymorphisms in the APOA gene are major determinants of plasma Lp(a) variation in American Samoans.     

Elevated CO22 and Leaf Longevity in the C44 Grassland-Dominant Andropogon gerardii

In central U.S. grasslands, plant and ecosystem responses to elevated CO2 are most pronounced when water availability is limited. In a northeast Kansas grassland, responses to elevated CO2 in leaf area, number, development, and longevity were quantified for the tallgrass prairie dominant, Andropogon gerardii. Plants were grown in open-top chambers (OTCs) modified to limit water availability and to maximize responses to elevated CO2. In OTCs with elevated (x2 ambient) levels of CO2, aboveground biomass production and leaf water potentials were increased significantly compared with those of plants in OTCs with ambient CO2. There were no differences in leaf area or leaf number per tiller in A. gerardii in elevated compared with ambient OTCs. However, leaf area in adjacent unchambered plots with greater water availability was significantly higher than in the OTCs. The time required for developing leaves to achieve maximum leaf area was reduced by 29%, and the period of time until leaves senesced was increased by 20% for plants exposed to elevated compared with ambient CO2. Thus, leaves of this C4 grass species expanded more rapidly (6 d) and remained green longer (9 d) when exposed to elevated CO2. Such CO2-mediated increases in leaf longevity in the dominant species may allow this grassland to respond more opportunistically to temporally variable rainfall patterns in high-CO2 environments. These responses should be included in leaf-based simulation models that attempt to mechanistically link physiological alterations to predicted canopy responses to increased CO2.     

Experimental inoculation of domestic pigeons (Columbia livia) and zebra finches (Poephila guttata) with Neospora caninum tachyzoites

Dogs are a definitive host of Neospora caninum and cattle are intermediate hosts. Alternative life-cycles have not been investigated. Foxes are frequently seropositive, but may not commonly prey upon cattle; therefore, other intermediate hosts may exist that are frequent prey of foxes. Three domestic pigeons (Columbia livia) and three zebra finches (Poephila guttata) were inoculated with N. caninum tachyzoites, to determine if they could serve as intermediate hosts. Tissue culture, PCR, serology, and histology were all positive for one or more pigeons. All finches resisted infection. Further testing of columbiform birds as intermediate hosts of N. caninum is warranted.     

Increased cytochrome P450 17alpha-hydroxylase promoter function in theca cells isolated from patients with polycystic ovary syndrome involves nuclear factor-1

Cytochrome P450 17alpha-hydroxylase (CYP17) gene expression and androgen biosynthesis are persistently elevated in theca cells isolated from ovaries of women with polycystic ovary syndrome (PCOS). We previously reported that -235 to -109 bp of the CYP17 promoter confers increased CYP17 promoter function in PCOS theca cells. In this report, additional deletion and mutational analyses of the CYP17 promoter were performed to identify the sequences that contribute to increased CYP17 promoter function in PCOS theca cells. Results of these analyses established that augmented promoter function in PCOS theca cells results from preferentially increased basal regulation conferred by sequences between -188 and -147 bp of the CYP17 promoter. Scanning mutant analysis demonstrated that mutations within a 16-bp sequence, spanning -174 to -158 bp of the promoter, ablated increased basal CYP17 promoter function in PCOS theca cells. EMSA analysis demonstrated that the NF-1 family member, NF-1C, bound this sequence. Cotransfection of several NF-1C isoforms expressed in normal and PCOS cells repressed CYP17 promoter function. NF-1C protein and DNA binding were reduced in PCOS theca cell nuclear extracts, as compared with normal. Another NF-1C site between -102 and -90 bp of the promoter was also identified. However, mutation of this site had no effect on differential promoter function in PCOS theca cells. These studies demonstrate that 1) augmented CYP17 promoter function in PCOS theca cells results from increased basal regulation, and 2) diminished NF-1C-dependent repression may be one mechanism underlying increased basal CYP17 promoter activity and altered gene expression in PCOS theca cells.     

TAT-mediated protein transduction into mammalian cells

Manipulation of mammalian cells has been achieved by the transfection of expression vectors, microinjection, or diffusion of peptidyl mimetics. While these approaches have been somewhat successful, the classic manipulation methods are not easily regulated and can be laborious. One approach to circumvent these problems is the use of HIV TAT-mediated protein transduction. Although this technology was originally described in 1988, few improvements were reported in the subsequent 10 years. In the last few years, significant steps have been taken to advance this technology into a broadly applicable method that allows for the rapid introduction of full-length proteins into primary and transformed cells. The technology requires the synthesis of a fusion protein, linking the TAT transduction domain to the molecule of interest using a bacterial expression vector, followed by the purification of this fusion protein under either soluble or denaturing conditions. The purified fusion protein can be directly added to mammalian cell culture or injected in vivo into mice. Protein transduction occurs in a concentration-dependent manner, achieving maximum intracellular concentrations in less than 5 min, with nearly equal intracellular concentrations between all cells in the transduced population. Full-length TAT fusion proteins have been used to address a number of biological questions, relating to cell cycle progression, apoptosis, and cellular architecture. Described here are the fundamental requirements for the creation, isolation, and utilization of TAT-fusion proteins to affect mammalian cells. A detailed protocol for production and transduction of TAT-Cdc42 into primary cells is given to illustrate the technique.     

Effects of immunization against GnRH, melengestrol acetate, and a trenbolene acetate/estradiol implant on growth and carcass characteristics of beef heifers

A 2 x 3 factorial experiment was conducted to determine the effects of an implant (trenbolene acetate/estradiol or no implant) and method of estrus suppression (immunization against GnRH, melengestrol acetate, or no suppression) on growth performance and carcass characteristics of heifers fed for slaughter. At the start of a 21-d feed adaption phase, crossbred beef heifers (n = 144, 390+/-2.8 kg) were given their first dose of an anti-GnRH vaccine or started on melengestrol acetate (MGA). Thereafter, heifers were fed a high-concentrate diet (78% barley grain) for 84 d (Days 0 to 83), received implants on Day 0, a second vaccination on Day 21, and were slaughtered on Days 84 or 85. Implanting increased average daily gain (1.72 vs 1.50 kg/d, P < 0.01), feed efficiency (6.02 vs 6.75 kg dry matter intake/kg gain, P < 0.01), preslaughter weight (532 vs 513 kg, P < 0.01), carcass weight (301 vs 289 kg, P < 0.01), and ribeye area (88.6 vs 85.9 cm2, P < 0.05), but had no affect (P > 0.05) on dry matter intake, grade fat thickness, marbling score, or lean yield. Compared to heifers fed MGA, those immunized against GnRH had a greater ribeye area (90.0 vs 84.6 cm2) and lean yield (63 vs 61%), and had thinner grade fat (7.5 vs 8.6 mm; P < 0.05 for each). Furthermore, immunized heifers had lower (P < 0.001) plasma progesterone concentrations than control heifers on Days 42, 63 and 83. Heifers fed MGA had less estrus mounting activity (P < 0.05) and lower plasma progesterone concentrations (P < 0.001) than the remaining heifers. Method of estrus suppression did not affect (P > 0.05) preslaughter weight, average daily gain, dry matter intake, feed efficiency, carcass weight, or marbling score. In conclusion, implanting significantly increased growth performance and preslaughter and carcass weights. Compared to heifers fed MGA, immunization against GnRH significantly increased ribeye area and lean yield, and reduced grade fat thickness