Beyond overlap: Considering habitat preference and fitness outcomes in the umbrella species concept.

Umbrella species and other surrogate-species approaches to conservation provide an appealing framework to extend the reach of conservation efforts beyond single species. For the umbrella species concept to be effective, populations of multiple species of concern must persist in areas protected on behalf of the umbrella species. Most assessments of the concept, however, focus exclusively on geographic overlap among umbrella and background species, and not measures that affect population persistence (e.g., habitat quality or fitness). We quantified the congruence between the habitat preferences and nesting success of a high-profile umbrella species (greater sage-grouse, Centrocercus urophasianus, hereafter "sage-grouse"), and three sympatric species of declining songbirds (Brewer's sparrow Spizella breweri, sage thasher Oreoscoptes montanus, and vesper sparrow Pooecetes gramineus) in central Wyoming, USA during 2012 - 2013. We used machine-learning methods to create data-driven predictions of sage-grouse nest-site selection and nest survival probabilities by modeling field-collected sage-grouse data relative to habitat attributes. We then used field-collected songbird data to assess whether high-quality sites for songbirds aligned with those of sage-grouse. Nest sites selected by songbirds did not coincide with sage-grouse nesting preferences, with the exception that Brewer's sparrows preferred similar nest sites to sage-grouse in 2012. Moreover, the areas that produced higher rates of songbird nest survival were unrelated to those for sage-grouse. Our findings suggest that management actions at local scales that prioritize sage-grouse nesting habitat will not necessarily enhance the reproductive success of sagebrush-associated songbirds. Measures implemented to conserve sage-grouse and other purported umbrella species at broad spatial scales likely overlap the distribution of many species, however, broad-scale overlap may not translate to fine-scale conservation benefit beyond the umbrella species itself. The maintenance of microhabitat heterogeneity important for a diversity of species of concern will be critical for a more-holistic application of the umbrella species concept.

Microfaunal remains at Middle Pleistocene Qesem Cave, Israel: Preliminary results on small vertebrates, environment and biostratigraphy.

Preliminary results of the investigation of the microfauna at the Acheulo-Yabrudian Middle Pleistocene site of Qesem Cave, Israel, are presented. Thus far the assemblage includes ca. 10,000 bone and tooth fragments, of which 50% could be identified to the generic and some hundreds to the species level. Based on the current material, the fauna includes the following squamate reptiles: Laudakia sp., Chamaeleo sp., Gekkonidae indet., Lacertidae indet., Scincidae indet., Pseudopus sp., Varanus sp., Colubroidea indet. (at least three species) and micromammals: Suncus etruscus, Crocidura cf. leucodon, Crocidurinae indet. (large form), Chiroptera indet., Sciurus cf. anomalus, Cricetulus cf. migratorius, Microtus guentheri, Nannospalax ehrenbergi, Dipodillus cf. dasyurus, Meriones cf. tristrami, Gerbillidae indet., Mus cf. musculus, Apodemus cf. flavicollis. These results suggest that the fauna includes only taxa that occur recently in the territory of Israel. The ecological preferences of the nearest living relatives of the recorded taxa allow us to infer a paleoenvironment with a mosaic of open and woodland habitats. However, comparing the lower with the upper levels of the microfauna-bearing profile, a slight shift towards more wooded conditions might be detectable. Biostratigraphical inferences from the recorded micromammal taxa cover a rather wide age range, whereas the radiometric (U-series and preliminary TL) dating enable a provisionally estimated date for the microfauna-bearing levels at 360-300 ka. Detailed morphometric comparisons with material from other sites in the region are necessary and may yet provide further insights.

Aluminum-induced calcium deficiency syndrome in declining red spruce.

Prolonged suppression of cambial growth has apparently caused a decline in radial growth in many mature red spruce, Picea rubens. Surveys indicate that this decline occurs in trees throughout the natural range of red spruce and is independent of elevation, tree size, and age class. In addition, crowns of mature red spruce at high elevations across the northeastern United States have been dying back. Understanding the physiological basis for the growth decline is essential for the judicious management of the red spruce resource. A sequence of events is inferred through which an imbalance of aluminum and calcium in the fine root environment reduces the rate of wood formation, decreases the amount of functional sapwood and live crown, and leaves large trees more vulnerable to extant secondary diseases and insect pests.

Timing of peak bone mass in Caucasian females and its implication for the prevention of osteoporosis. Inference from a cross-sectional model.

To determine the timing of peak bone mass and density, we conducted a cross-sectional study of bone mass measurements in 265 premenopausal Caucasian females, aged 8-50 yr. Bone mass and bone mineral density were measured using dual X-ray absorptiometry and single-photon absorptiometry at the spine (anteroposterior, lateral), proximal femur, radius shaft, distal forearm, and the whole body. Bone mass parameters were analyzed using a quadratic regression model and segmented regression models with quadratic-quadratic or quadratic-linear form. The results show that most of the bone mass at multiple skeletal locations will be accumulated by late adolescence. This is particularly notable for bone mineral density of the proximal femur and the vertebral body. Bone mass of the other regions of interest is either no different in women between the age of 18 yr and the menopause or it is maximal in 50-yr-old women, indicating slow but permanent bone accumulation continuing at some sites up to the time of menopause. This gain in bone mass in premenopausal adult women is probably the result of continuous periosteal expansion with age. Since rapid skeletal mineral acquisition at all sites occurs relatively early in life, the exogenous factors which might optimize peak bone mass need to be more precisely identified and characterized.

Association of bovine papillomavirus type 2 and bracken fern with bladder cancer in cattle.

The bladder cancer syndrome that often accompanies chronic enzootic hematuria in cattle grazing on pastures infested by bracken fern has been experimentally reproduced in animals fed a diet of bracken. The experimentally induced tumors were histologically and pathologically indistinguishable from the naturally occurring ones and comprised two main types: (a) carcinoma of the urothelium identical to that seen in humans; and (b) hemangioendotheliomas of the subjacent capillaries. Often the two types of tumor occurred together in the same bladder. In animals experimentally immunosuppressed with azathioprine "bracken type" hemangiomas developed in the bladder lining. DNA of bovine papillomavirus (BPV) type 2 was found in 46% (7 of 15) of the natural cancer cases and in 69% (9 of 13) of the experimentally induced lesions, independently of histological type and including the hemangiomas of the azathioprine-treated animals, suggesting a close association between BPV and bovine bladder neoplasia. Moreover, BPV-2 DNA was found in experimental animals that had not been inoculated with BPV at all or had been inoculated with a different BPV type and had been kept in isolation, suggesting that BPV can persist in a latent state and be activated when the animal is exposed to the bracken cocarcinogens and to immunosuppressants.

Amplification of specific DNA sequences in C127 mouse cells transformed by bovine papillomavirus type 4.

A rearranged bovine papillomavirus type 4 DNA fragment, present in a line of transformed C127 cells, was molecularly cloned into lambda GT 10. The rearranged viral fragment was found to consist of two separate sequences of 3.8 kb each. Clone A comprised 3.3 kb of host DNA linked to 0.5 kb of BPV-4 DNA. Clone B consisted of 2.2 kb of host DNA linked to 1.5 kb of BPV-4 DNA. The AB locus was found to be amplified and rearranged in a number of different BPV-4 transformed C127 cell lines, irrespective of the presence of viral DNA. The rearranged amplified locus was transferred and maintained in second and third round transformants. We propose that the altered AB locus is involved in the maintenance of the transformed phenotype in the absence of the BPV-4 DNA, and may act as an activated oncogene.

BPV-4 induces amplification and activation of 'silent' BPV-1 in a sub-line of C127 cells.

BPV-4-induced malignant transformation of C127 mouse fibroblasts in vitro is the result of a 'hit and run' mechanism. Viral DNA is lost on continued subculture of transformed cell lines without loss of any malignant characteristics. The DNA from these cells harbours specific amplified sequences. Two such amplified fragments of approximately 10 kb and 12 kb were molecularly cloned and designated HL-10 and HL-12 respectively. HL-10 transformed C127 cells efficiently and therefore encodes a transforming function, whereas the 12 kb clone did not. Heteroduplexes showed that HL-12 was homologous to HL-10 except for two additional tandem copies of an approximately 1.7 kb sequence. Sequence analysis of HL-10 revealed that the clone contained a 5.2 kb region from BPV-1 including the transforming ORFs. Transformation studies have shown differences between HL-10 and BPV-1, indicating that the host flanking sequences may contribute to the transforming potential of the BPV-1 ORFs. The BPV-1 DNA was associated with sequences homologous to murine autonomously replicating sequences (ARS) implicated in the establishment of multiple tandem DNA repeats. As the parental cells contain the set of sequences amplified in transformed cells in single copy and show none of the characteristics of transformed cells, we conclude that BPV-4 has activated these sequences by amplification and rearrangement. These phenomena may be mediated through an interaction between BPV-4 proteins and the BPV-1 origin of DNA replication or via the ARS region.

Coordinate and non-coordinate accululation of aspartate transcarbamylase and dihydroorotase in synchronous Chlorella cells growing on different nitrogen sources.

Regulation of the levels of aspartate transcarbamylase (carbamoylphosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2) and dihydroorotase (L-5,6-dihydro-orotate amidohydrolase, EC 3.5.2.3) was studied in synchronous cultures of the eucaryotic microorganism Chlorella. Analytical polyacrylamide gel electrophoresis and sucrose density-gradient centrifugation studies revealed that these cells contain a single aspartate transcarbamylase and a dihydroorotase with apparent molecular weights of 160 000 and 80 000, respectively. In synchronous cells cultured in nitrate medium, these two enzymes accumulated in single step-patterns over different periods of the cell cycle. In contrast, these enzymes accumulated in a coordinate manner throughout the cell cycle in ammonium medium. Experiments with inhibitors of protein and RNA synthesis indicated that dihydroorotase is stable in vivo and suggested that cell cycle changes in the turnover rate of aspartate transcarbamylase might determine whether or not these enzymes accumulate in a coordinate manner. Although uracil and uridine could be absorbed and metabolized by the cells, synthesis of these two enzymes could not be repressed by culturing synchronous cells in medium, containing high concentrations (29-40 mM) of uracil or uridine, for an entire cell cycle.

Zinc-binding properties of bovine and human milk in vitro: influence of changes in zinc content.

The total zinc content of samples of fat-free human and bovine milk was 3.5 and 3.6 micrograms/ml, respectively. The total protein content was 5.3 and 29.0 mg/ml, respectively, Sephadex G-75 gel filtration chromatography of bovine milk revealed no evidence of zinc associated with low molecular weight fractions (less than 2000 daltons). However, chromatography revealed that 10% of the zinc in human milk was associated with these fractions. When the zinc content of these milks was raised by adding Zn2+ in vitro, chromatography of milk from both species revealed the presence of zinc with the low molecular weight fractions. More zinc was associated with these fractions from human milk than from bovine milk. Recovery of zinc was nearly 100% for all columns. It is proposed, based upon these binding data, that the association of zinc with low molecular weight components of milk is related in part to both protein content and composition and the relative zinc concentrations. These binding differences might influence the bioavailability of zinc from milk.

Spinal bone density and calcium intake in healthy postmenopausal women.

Dietary calcium intake and bone mineral density (BMD) of the lumbar spine (L2-L4) were determined in 131 healthy free-living postmenopausal women (aged 64.7 +/- 7.6 y, means +/- SD). The calcium consumption for the total population was 606 +/- 302 mg/d. Subjects consuming less than the population mean of dietary calcium had significantly lower BMDs than did subjects with intakes above the mean (P less than 0.009); these two groups did not differ in basic demographic characteristics. Additional analyses using a stepwise univariate regression model demonstrated that BMD was significantly associated with body weight (P less than 0.001) and dietary calcium intake (P less than 0.02). These data support the hypothesis that dietary calcium intake is a determinant of skeletal health in postmenopausal women.

Meal effects on calcium absorption.

The effect of coingestion of a meal on calcium absorption efficiency was investigated using Ca carbonate, milk, and a new Ca delivery system, CCM. Six experiments with 168 rats and two experiments with 46 normal young women showed that coingestion of a light meal of varied composition enhanced absorption efficiency from all three sources by 10-30% above levels achieved when the test load was ingested without other foods. The differences taken as a whole were highly statistically significant.

Effects of different calcium sources on iron absorption in postmenopausal women.

We measured the effect of calcium from food and supplement sources on whole-body retention of 59Fe in 19 normal postmenopausal women. Each woman received a placebo and 500 mg calcium from a mixed calcium citrate-malate salt (CCM), from orange juice plus CCM, and from milk after a test breakfast meal to which 59Fe had been added. The test meal contained 238 mg calcium. Whole-body countings of 59Fe were performed before and 30 min and 2 wk after each test meal. Retention of 59Fe was 8.3 +/- 1.1% (means +/- SEM) with placebo, 3.4 +/- 0.78% with milk, 6.0 +/- 0.97% with CCM, and 7.4 +/- 1.7% with CCM plus orange juice. When compared with placebo, milk and CCM significantly lowered iron retention (p less than 0.05) whereas CCM plus orange juice did not. The reduction with milk was greater than that with CCM (p less than 0.05) or CCM plus orange juice (p less than 0.05). The differences in the effects of these calcium sources on 59Fe retention may result from their varied contents of citric and ascorbic acids, known enhancers of iron absorption.

High throughput detection of retrovirus-associated reverse transcriptase using an improved fluorescent product enhanced reverse transcriptase assay and its comparison to conventional detection methods.

The development and application of a novel, sensitive TaqMan fluorescent probe-based product enhanced RT test (F-PERT) for the detection of retrovirus are described. The assay allows discrimination between the amplification signals generated by genuine positive signals that result from retroviral RT activity and the RT-like activity from DNA polymerases. The RT-like activity from DNA polymerases was suppressed by the addition of activated calf-thymus DNA with no reduction in the RT activity. A linear relationship between threshold cycle (C(T)) and the number of virus particles was demonstrated, allowing quantification of retroviruses in unknown samples. The F-PERT assay was able to detect a wide range of retroviral RT activities, including that from porcine endogenous retrovirus (PoERV), murine leukaemia virus (MLV), simian foamy virus (SFV), simian immunodeficiency virus (SIVmac) and squirrel monkey retrovirus (SMRV). The detection limit of SMRV, MLV and PoERV was approximately 100 virion particles and the test was able to detect at least 10(2) molecules of purified RT enzyme. RT activity was not detected in cellular lysates and supernatants from MRC-5, BT, VERO, or Raji cells, whereas RT activity was detected in C1271, Mus dunni, K-Balb, BHK-21, CHO-K1, SP2/0-Ag14 and NSO cell supernatants. RT activity was also detected in the Spodoptera cell line Sf9.

Comparison of electron microscopic techniques for enumeration of endogenous retrovirus in mouse and Chinese hamster cell lines used for production of biologics.

Murine myeloma and Chinese hamster ovary cells are used widely in the manufacture of recombinant proteins for biopharmaceuticals. However, rodent cell lines express endogenous retrovirus, which necessitates appropriate design of purification processes to remove virus in excess of the calculated maximum retroviral load. Currently, electron microscopy is the method of choice for determination of retroviral titre in bulk harvest. In this study we compared three electron microscopy techniques to determine retroviral titre in bulk harvest. These were direct negative stain, negative stain after sucrose-density purification and thin section electron microscopy of pelleted supernatant. The study demonstrated that the level of C-type retrovirus associated with cells was predictive of the viral load in cell culture supernatants. The most accurate method for quantifying viral load was direct counting, followed by thin section of pelleted supernatant and negative stain after sucrose concentration. The most practical method was thin section of resuspended pelleted supernatant, which gave improved detection limits.

Design and validation of immunological tests for the detection of Porcine endogenous retrovirus in biological materials.

The present study details the design and demonstrates function for a series of reagents and methods to allow the detection of exposure to antigens specific for Porcine endogenous retrovirus (PERV). The detection of PERV is carried out by the means of a variety of immunological screening methods including, indirect immunofluorescence, Western blotting and enzyme linked immunosorbent assay (ELISA) for the detection of antibodies in serum specific for PERV gag and env antigens. Alternatively, PERV-specific antisera for gag and env can be used to detect viral antigen in serum or other samples. PERV env peptides with potential specificity for the known PERV types are also described. Antisera against the peptides can be used to detect PERV antigens directly or to characterise viral type. Using electron microscopy coupled with labelled PERV-gag-specific antisera it was possible to visualise PERV virions.

Computed tomographic imaging of the coronary arterial tree-use of local tomography.

It has been demonstrated previously (see E.L. Ritman and A.A. Bove, in State of the Art in Quantitative Coronary Arteriography, p.67-78, 1986) that coronary artery anatomy can be visualized using high-speed, volume-scanning X-ray CT (computed tomography). In the current study it is demonstrated that local image reconstruction (i.e. the reconstruction kernel is ~2(+) mm long), as distinct from more conventional global image reconstruction (i.e. 200(+) mm kernel length), has the advantage of reducing the need for operator interactive image processing. In addition, the local reconstruction algorithm eliminates the need for recording the X-ray projection data over the full transaxial extent of the thorax because it requires only the X-ray attenuation data over the region of the heart. This latter aspect reduces the dynamic range requirements for the sensors and could reduce total X-ray exposure.

The biology of papillomaviruses and their role in oncogenesis.

A large number of different papillomavirus types have been identified. Several viruses often infect the same species and each virus is associated with a defined tissue. Recent evidence has shown that certain benign lesions can undergo malignant transformation in both animals and humans in response to genetic or environmental factors. Fine mapping of the structure and function of the viral genome may enhance our understanding of the interaction between the virus and the cofactors involved in malignant transformation.

Papillomaviruses and their involvement in oncogenesis.

Papillomaviruses have been identified as the causative agents of benign epithelial proliferation in many animals including man. Recent evidence has shown that each viral type will only infect a specific species and tissue. Furthermore, certain papillomavirus types have been found associated with lesions capable of malignant conversion, particularly in response to secondary physical or chemical factors. Many advances have been made in identifying particular papillomavirus types inducing papillomas capable of progression and in the identification of the virally encoded functions involved in viral replication and cellular transformation.

General and histopathological results of a two-year study of rats fed semi-purified diets containing casein and soya protein.

Two semi-purified diets, identical except for protein (soya or casein), and a Purina chow diet were fed to groups of Sprague-Dawley rats and compared over a 22-month period for effects on body weight, organ weight, feed consumption, feed efficiency, protein efficiency, organ-to-body-weight ratios, certain organ mineral levels, gross pathology and histopathology. Feed efficiency, protein efficiency, body weight, and most organ weights were higher in the groups fed soya and casein, while feed consumption and most organ-to-body-weight ratios were relatively lower. Significant increases in the final pathology findings in the groups fed soya and casein included nephrocalcinosis (in females), hepatocellular vacuolization, gastric trichobezoars (hairballs), ulcerative pododermatitis (hock ulcers) and splenomegaly. This study demonstrates that the long-term feeding of semi-purified diets is feasible. Moreover, the differences observed between the protein sources, that is, soya and casein, were minimal, which suggests that either protein source is acceptable for long-term feeding studies.

Intraduodenal delivery of intrinsically and extrinsically labelled CaCO3 in the rat: effect of solubilization on calcium bioavailability.

The dissolution of CaCO3 before intraduodenal administration was found to be an important factor determining calcium (Ca) bioavailability. Extrinsically and intrinsically labelled 47CaCO3 preparations were sequentially dissolved by serial additions of HCl. Aliquots of these preparations were collected before (no HCl added) and during the solubilization process and administered intraduodenally to rats. Whole body 47Ca retention 72 h post-dose was used as a measure of Ca bioavailability. Although dissolution of CaCO3 significantly increased Ca bioavailability (P < 0.001), Ca from both intrinsically and extrinsically labelled CaCO3 was absorbed and retained to some extent without prior acid dissolution. Due to a disproportionately high concentration of 47Ca on the particle surface, extrinsically labelled 47CaCO3 overestimated bioavailability when unsolubilized or partially solubilized CaCO3 preparations were used (P < 0.05). These data indicate that dissolution is a determining factor for Ca bioavailability from CaCO3. Incomplete dissolution will significantly limit but not completely prevent Ca bioavailability. The disintegration and dissolution characteristics of commercial CaCO3 preparations, which vary widely, may produce important differences in Ca absorption.