PubMed Health⌕ Search

Biomedical subjects

A Perkins

Publications and source records attributed to A Perkins.

At least 55 records · Page 3Linked to original sources
PubMed1992

Luteinizing hormone, testosterone, and behavioral response of male-oriented rams to estrous ewes and rams.

During the breeding season three experiments were conducted to evaluate the LH and testosterone (T) response of rams with male sexual orientation (e.g., male-oriented homosexual rams) to female sheep, to male sheep, and to treatment with LHRH. Male-oriented rams were identified through a series of sexual performance and sexual preference tests. Treatments included exposure to estrous females and to males for 15 min (Exp. 1) and exposure to estrous females and to males for 8 h (Exp. 2). Behavioral responses to stimulus animals were recorded. In Exp. 2 homosexual rams mounted males more than females (P less than .02) and exhibited more flehmen (P less than .002) and investigatory sniffs (P less than .01) when exposed to males vs females. Acts of aggression (butting the stimulus animals) did not differ by gender (P greater than .1). Flehmen and butting were positively correlated to LH secretion (P less than .02) of rams exposed to females but not to males. In Exp. 1, LH concentration determined every 15 min for 6 h was not affected (P greater than .05) by the gender of the stimulus animal. In Exp. 2, LH pulse frequency and concentration were similar (P greater than .05) by treatment. Lack of an LH response to sexual activity in homosexual rams was not a result of pituitary or gonadal insensitivity; within 1 h of a single injection of LHRH both LH and T increased (Exp. 3).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Luteinizing hormone and testosterone response of sexually active and inactive rams.

The objective of this study was to identify rams exhibiting high (HP) and low (LP) levels of sexual performance and to determine whether their respective behavioral responses to ewes in estrus were related to changes in serum testosterone (T) and LH concentrations. Rams were selected on the basis of standardized serving capacity tests. Plasma T and LH concentrations in rams were measured in three experiments: 1) after 15 min of exposure to estrous ewes, 2) after an injection of 500 ng of LHRH, and 3) during an 11-h exposure to estrous ewes. During 15 min of exposure to ewes, HP rams were sexually active, whereas LP rams showed no sexual interest. Secretion of LH was similar (P greater than .05) between ram groups. Sexual arousal, copulation, and ejaculation of HP males were not related (P greater than .05) to LH secretion. Exposure to estrous ewes for 11 h, however, stimulated LH pulse frequency and elevated basal LH and T concentrations in HP but not LP rams (P less than .001). Luteinizing hormone secretion was positively correlated to the frequency of mounts (r = .19; P less than .01) and ejaculation (r = .17; P less than .03). Aggressive behavior of rams directed at ewes was negatively correlated to LH (r = -.22 P less than .003). Concentrations of LH and T after LHRH injection were similar between HP and LP rams (P greater than .05). These results show that the effects of the ewe on LH secretion of rams depend on length of the exposure period and sexual activity of the male.

Aggression↗
PubMed1992

Sexual performance of rams in serving capacity tests predicts success in pen breeding.

The purpose of this study was to determine the extent to which sexual performance (serving capacity) tests can be used to predict the sexual behavior and reproductive success of rams in the context of pen mating. Standard serving capacity tests were used to select four low (LP) and four high-performing (HP) rams from a population of 94 yearling males. Each selected ram was then exposed to approximately 30 estrus-synchronized ewes for a 9-d period. Ejaculations observed and mounting marks left on ewes confirmed the greater (P less than .001) sexual activity of the HP rams in the field. In addition, ewes exposed to HP rams had a higher lambing percentage, more lambs born, and more live lambs born per ewe. Ram classification was not related (P greater than .60) to the number of lambs born per ewe lambing (prolificacy). It was concluded that serving capacity tests, properly conducted, can be used to predict ram mating performance and thus aid in establishing more efficient ram-to-ewe stocking rates.

Animals↗
PubMed1991

Concomitant delineation of surface Ig, B-cell differentiation antigens, and HLADR on lymphoid proliferations using three-color immunocytometry.

Accurate and consistent enumeration of B-cell subpopulations in lymphoid tissue was achieved through multiparameter three-color immunofluorescence and flow cytometric analysis (FCM). Phycoerythrin (PE)-anti-CD19 (Leu12) and biotinylated anti-HLADr/streptavidin-Duochrome (PE/Texas Red), used in conjunction with polyclonal fluorescein isothiocyanate (FITC) conjugated anti-surface immunoglobulin (SIg) antibodies, effectively separated non-specific binding and background fluorescence from true B-cell surface FITC immunofluorescence, while concomitantly analyzing for HLADr and CD19 phenotypic expression/deletion. Autofluorescence was measured to establish a fluorescence threshold. A second control measured non-specific binding of isotypic control mouse Ig and non-immune rabbit IgG. Cell suspensions from 128 samples of various lymphoid proliferations were studied. In 116 of the 128 samples, kappa/lambda ratios determined by flow cytometry correlated well with immunocytology results obtained using cytospins from the same cell suspension and with histopathologically established diagnosis. Clonality and lineage as defined immunotypically by flow cytometry was concordant with genotypic results in 64 of the 67 cases evaluated. SIg, HLADr, and CD19 deletions were demonstrated by flow cytometry in 8, 4, and 1 case(s), respectively. Discordance was usually attributable to selective loss of large neoplastic cells in flow cytometry specimens or absent expression of SIg by some cytoplasmic Ig (CIg+) lymphomas.

Antigens, CD↗
PubMed1991

The diphosphonate space: a useful quantitative index of disease activity in patients undergoing hydroxy methylene diphosphonate (HMDP) bone imaging for Paget's disease [corrected].

Comparison between the plasma levels of intravenously injected technetium 99m hydroxy methylene [corrected] diphosphonate (99mTc-HMDP) and chromium 51 ethylene diamine tetra-acetic acid (51Cr-EDTA) reflects the uptake of diphosphonate into bone (the diphosphonate space). This can be used as an index of skeletal function in metabolic bone disease. In a series of 49 patients with Paget's disease the diphosphonate space (DPS) correlated well with other indicators of disease activity such as alkaline phosphatase and urinary hydroxyproline levels. The DPS is a good predictor of the volume of skeletal involvement as estimated from bone images. The DPS also provides a sensitive indicator of response to treatment with intravenously administered bisphosphonate. The DPS is simple to perform and is a useful adjunct to routine bone imaging.

Alkaline Phosphatase↗
PubMed1991

A turnover prevention program for child care workers: a failed effort?

This study describes efforts to reduce turnover of child care workers, one of the many activities undertaken within the Milieu Enhancement Project. This 3-year endeavor, funded by the Division of Nursing of the Department of Health and Human Services, allowed nurses to provide care for orphaned and disadvantaged children, test the value of a nursing practice model, analyze the cost-benefit and the cost-effectiveness of the project, computerize existing and model-derived health care records, and allow faculty and graduate students to practice autonomously in a nontraditional setting. The research question was: What is causing job dissatisfaction in child care workers? Nurses and social workers used the Milieu Enhancement Model to conceptualize, address, and evaluate factors contributing to turnover. While statistical significance was not obtained in the turnover rates before and after implementation of the Turnover Prevention Program, the milieu was enhanced in many ways. Data are presented to support many aspects of the program's success.

Adult↗
PubMed1991

Isolation from human placental extracts of a preparation possessing 'early pregnancy factor' activity and identification of the polypeptide components.

Protocols are described for the isolation from human placental extracts of a preparation which is active in the rosette inhibition assay by inducing an increased rosette inhibition titre. In this respect the preparation mimics the effects of pregnancy sera, an ability ascribed to the presence in these sera of a so-called 'early pregnancy factor' (EPF). In addition to this activity in the rosette inhibition assay, the preparation was also shown to modulate the expression of cell surface immunoglobulin on peripheral blood mononuclear cells. The polypeptide composition of the preparation was relatively simple, as revealed by SDS-PAGE. There was a major 12 kd polypeptide previously isolated in a rosette inhibition titre-active preparation from ovine placental extracts, and a small amount of 68 kd polypeptide. N-Terminal amino acid sequence analyses obtained after blotting onto polyvinylidene difluoride membranes identified the latter polypeptide as serum albumin and the major 12 kd polypeptide as human thioredoxin. Gel permeation analysis partially resolved activity expression away from the 12 kd polypeptide as activity expression was also found to be associated with low mol. wt material. It is concluded that EPF activity expression in pregnancy sera involves molecules related to and associated with thioredoxin.

Amino Acid Sequence↗
PubMed1991

Structural and functional analysis of the human immunodeficiency virus type 2 Rev protein.

The Rev proteins of the human immunodeficiency virus (HIV) are necessary for expression of viral structural gene products. Site-directed mutations were made within the HIV-2 rev gene to identify functional domains. We observed that similar to HIV-1 Rev, the HIV-2 Rev protein was phosphorylated, albeit to a much lesser extent than was HIV-1 Rev. We also found that like HIV-1 Rev, HIV-2 Rev localized to the nucleus, with a marked accumulation in the nucleolus. Mutations within a stretch of basic residues prevented both nuclear and nucleolar localization. Furthermore, mutant Rev proteins able to localize in the nucleus but unable to localize in the nucleolus were nonfunctional.

Amino Acid Sequence↗
PubMed1991

The HIV-1 Rev protein: a model system for coupled RNA transport and translation.

The impact of the Rev protein of the human immunodeficiency virus type 1 (HIV-1) on RNA transport, intranuclear RNA distribution, and gene expression was examined for two Rev-dependent expression systems by means of fluorescence in situ hybridization, immunofluorescence, S1 nuclease protection, and functional assays. In the pgTat expression system, which utilizes authentic HIV-1 splice signals, unspliced mRNA remained entrapped in the nucleus in the absence of Rev and was exported to the cytoplasm in its presence, consistent with published findings. In the pSVAR expression system, significant levels of mRNA were found in the nucleus and cytoplasm in both the presence and absence of Rev, but only in the presence of Rev was mRNA translated into protein. The presence of cytoplasmic untranslated mRNA in the absence of Rev was demonstrated by in situ hybridization analysis of individual cells as well as by S1 nuclease analysis of cell populations. The results indicate that Rev has the potential to affect translation as well as transport, suggesting the possibility that cellular mechanisms exist whereby the translational efficiency of an mRNA may be affected by the manner in which it is transported from the nucleus. Fluorescence hybridization also provided high-resolution visualization of the intranuclear distribution of RNAs containing the Rev response element. This demonstrated for both expression systems that mRNA was not highly localized in tracks or around the nucleolus in the presence or absence of Rev, a nucleolar protein, but was more widely distributed throughout the nucleus. In pgTat transfectants, HIV-1 RNA often became localized in 5 to 20 discrete large intranuclear clusters in the presence of Rev, the potential significance of which is discussed.

Biological Transport↗
PubMed1990

A cDNA for a protein that interacts with the human immunodeficiency virus Tat transactivator.

The human immunodeficiency virus (HIV) tat protein (Tat) is a positive regulator of virus gene expression and replication. Biotinylated Tat was used as a probe to screen a lambda gt11 fusion protein library, and a complementary DNA encoding a protein that interacts with Tat was cloned. Expression of this protein, designated TBP-1 (for Tat binding protein-1), was observed in a variety of cell lines, with expression being highest in human cells. TBP-1 was localized predominantly in the nucleus, which is consistent with the nuclear localization of Tat. In cotransfection experiments, expression of TBP-1 was able to specifically suppress Tat-mediated transactivation. The strategy described may be useful for direct identification and cloning of genes encoding proteins that associate with other proteins to modulate their activity in a positive or negative fashion.

ATPases Associated with Diverse Cellular Activitie↗
PubMed1990

Leukotrienes are active in the rosette inhibition assay mimicking the action of 'early pregnancy factor'.

Lipoxygenase metabolites have been found to be active in the rosette inhibition assay by inducing increased rosette inhibition titres. Leukotrienes B4, C4, D4 and E4 were identified as possessing this activity. The cycloxygenase products, prostaglandins E2, D2 and F2 alpha had no such activity; however, prostaglandin E2 and to a lesser degree prostaglandin D2 could counteract the activity of the leukotrienes in this assay. The identified leukotrienes are the first characterised molecules known to display activity in the assay. In this respect they mimic the action of early pregnancy serum, an action ascribed to a so called 'early pregnancy factor'.

Animals↗
PubMed1990

Flow cytometric competitive binding assay for determination of actinomycin-D concentrations.

A single step, separation free competitive binding reaction between the fluorescent antibiotic mithramycin and actinomycin-D for common binding sites on DNA coated 10 microns diameter microspheres is described. The fluorescence of the microspheres is measured with a flowcytometer. In the presence of a constant amount of mithramycin, the microsphere fluorescence is inversely proportional to actinomycin-D concentration.

Binding, Competitive↗
PubMed1990

Homeobox gene expression plus autocrine growth factor production elicits myeloid leukemia.

In the murine myelomonocytic leukemia WEHI-3B, proviral insertions have induced expression of the Hox-2.4 homeobox gene and the gene for the myeloid growth factor interleukin 3 (IL-3). To assess their potential oncogenic role, normal bone marrow cells were infected with retroviruses bearing the genes for IL-3 or IL-3 plus Hox-2.4. Unlike the IL-3 virus, the IL-3/Hox-2.4 virus was highly leukemogenic. Infected cells expressing both genes exhibited retarded differentiation in vitro, generated myelomonocytic cell lines, and provoked a rapid, transplantable myeloid leukemia in vivo. The oncogenic action of Hox-2.4 appears to derive from its ability to impede the IL-3-driven terminal differentiation of myeloid cells. The results suggest that homeobox genes can regulate key differentiation processes such as self-renewal capacity and that their inappropriate expression can be oncogenic.

Animals↗
PubMed1990

Identification of sequences important in the nucleolar localization of human immunodeficiency virus Rev: relevance of nucleolar localization to function.

The human immunodeficiency virus rev gene product regulates the expression of viral structural genes. It was recently shown that Rev regulates the export of viral structural mRNAs from the nucleus to the cytoplasm. Analysis of Rev subcellular localization reveals marked accumulation in the nucleolus, suggesting a role for the nucleolus in this export process. We report here the identification of amino acid residues critical to the nucleolar localization of Rev. Consistent with this finding, a Rev/beta-galactosidase fusion protein, harboring this region of Rev, localized entirely within the nucleolus. Of most significance, mutations that eliminated nucleolar localization markedly diminished Rev function, even though accumulation in the nucleoplasm was retained. These findings support a model whereby Rev-induced export of human immunodeficiency virus structural mRNAs from the nucleus to the cytoplasm is likely to involve nucleolar events.

Amino Acid Sequence↗
PubMed1990

Function of the human immunodeficiency virus types 1 and 2 Rev proteins is dependent on their ability to interact with a structured region present in env gene mRNA.

The interaction of the human immunodeficiency virus type 1 (HIV-1) Rev protein with a structured region in env mRNA (the Rev-responsive element [RRE]) mediates the export of structural mRNAs from the nucleus to the cytoplasm. We demonstrated that unlike HIV-1 Rev, which functions with both the HIV-1 and HIV-2 RREs, HIV-2 Rev functions only with the HIV-2 RRE. Rev-RRE binding studies suggested that the lack of nonreciprocal complementation stems from the inability of HIV-2 Rev to interact with HIV-1 RRE RNA. Maintenance of RNA secondary structure, rather than the primary nucleotide sequence, appeared to be the major determinant for interaction of both HIV-1 and HIV-2 Rev with the HIV-2 RRE. Moreover, the binding domain of the HIV-2 RRE recognized by HIV-1 Rev was dissimilar to the binding domain of the HIV-1 RRE, in terms of both secondary structure and primary nucleotide sequence. Our results support the hypothesis that function of HIV Rev proteins and possibly the functionally similar Rex proteins encoded by the human T-cell leukemia viruses (HTLVs) HTLV-I and HTLV-II is controlled by the presence of RNA secondary structure generated within the RRE RNA.

Base Sequence↗
PubMed1989

Effect of magnesium on depression of the monosynaptic reflex induced by 2-chloroadenosine or hypoxia in the isolated spinal cord of neonatal rats.

Superfusion of the isolated spinal cord of neonatal rats (4-9 days postpartum) with physiological medium containing 2-chloroadenosine (2-CA) or anoxic medium (equilibrated with 95% N2-5% CO2) depressed the evoked monosynaptic reflex (MSR) recorded extracellularly from a ventral spinal root. The effectiveness of 2-CA or anoxic medium in depressing the MSR was significantly reduced when the concentration of Mg2+ in the physiological medium was lowered from 1.25 X 10(-3) M to zero. The absence of Mg2+ resulted in a 7-fold shift to the right of the concentration-response curve to 2-CA and a reduction in the maximal depression of the MSR from 100% to 65 +/- 4% (mean +/- S.E.M.) of control. A 10 min exposure to anoxic medium containing 1.25 X 10(-3) M Mg2+ decreased the amplitude of the MSR to 23 +/- 6% of control, whilst in zero Mg2+ a decrease to only 50 +/- 5% of control was observed. These data provide further evidence that the response to adenosine, at the A1-receptor, is sensitive to Mg2+ ion concentration and suggest that there is an absolute requirement for Mg2+ in order to obtain full expression of the adenosine effect. Furthermore, the data are consistent with the hypothesis that adenosine is an important mediator of hypoxia-induced depression of the evoked MSR in the spinal cord, and suggest a potential role for Mg2+ during or after exposure to hypoxia in altering the actions of adenosine on neuronal activity or synaptic events.

2-Chloroadenosine↗