Evaluation in Appalachian pasture systems of the 1996 (update 2000) National Research Council model for weaning cattle.

This study was conducted to evaluate the accuracy of the National Research Council's (2000) Nutrient Requirements of Beef Cattle computer model when used to predict calf performance during on-farm pasture or dry-lot weaning and backgrounding. Calf performance was measured on 22 farms in 2002 and 8 farms in 2003 that participated in West Virginia Beef Quality Assurance Sale marketing pools. Calves were weaned on pasture (25 farms) or dry-lot (5 farms) and fed supplemental hay, haylage, ground shell corn, soybean hulls, or a commercial concentrate. Concentrates were fed at a rate of 0.0 to 1.5% of BW. The National Research Council (2000) model was used to predict ADG of each group of calves observed on each farm. The model error was measured by calculating residuals (the difference between predicted ADG minus observed ADG). Predicted animal performance was determined using level 1 of the model. Results show that, when using normal on-farm pasture sampling and forage analysis methods, the model error for ADG is high and did not accurately predict the performance of steers or heifers fed high-forage pasture-based diets; the predicted ADG was lower (P < 0.05) than the observed ADG. The estimated intake of low-producing animals was similar to the expected DMI, but for the greater-producing animals it was not. The NRC (2000) beef model may more accurately predict on-farm animal performance in pastured situations if feed analysis values reflect the energy value of the feed, account for selective grazing, and relate empty BW and shrunk BW to NDF.

Influence of mass of ruminal contents on voluntary intake and digesta passage in steers fed a forage and a concentrate diet.

To evaluate the influence of mass of ruminal contents on voluntary intake and ruminal function, five ruminally cannulated steers (550 kg) were fed an orchard grass hay diet ad libitum in a 5 x 5 Latin square experiment. The mass of ruminal contents was altered by adding varying weights of modified tennis balls to the rumen before the initiation of each 15-d experimental period. Treatments consisted of 50 balls with a specific gravity of 1.0, 1.1, 1.2, 1.3, or 1.4; the total weight of the balls was 7.45, 8.50, 9.25, 10.55, and 11.55 kg, respectively. Increasing the specific gravity of the balls added to the rumen decreased DMI and particle passage rate (P < 0.05) in a linear manner. A second experiment was conducted to evaluate the influence of mass of ruminal contents on voluntary intake and ruminal function of both forage and concentrate diets. Five ruminally cannulated steers (580 kg) were fed a 70% concentrate (DM basis) or an orchardgrass hay diet ad libitum in a 5 x 5 Latin square experiment. The mass of ruminal contents was altered as in the first experiment. Treatments consisted of 0 balls added to the rumen of steers fed concentrate diet (control), 75 balls with a specific gravity of 1.1 given to steers fed a concentrate diet, 75 balls with a specific gravity of 1.4 given to steers fed a concentrate diet, 75 balls with a specific gravity of 1.1 given to steers fed a hay diet, and 75 balls with a specific gravity of 1.4 given to steers fed hay diet. The addition of balls to the rumen of steers fed the concentrate diet decreased DMI (P < 0.05) compared with the 0-ball treatment, and increasing specific gravity of balls also decreased DMI (P < 0.01) for both concentrate and hay diets. Adding balls to the rumen of steers fed the concentrate diet decreased particle passage rate (P < 0.05), whereas increasing specific gravity of balls decreased particle passage rate for both concentrate and hay diet. The results of this study suggest that the density of ruminal digesta can have an influence on voluntary intake of both forage and concentrate diets.

The Health-Related Hardiness Scale: Spanish language equivalence and translation.

The article describes the development and initial evaluation of a Spanish language version of the Health-Related Hardiness Scale (HRHS) and reports on the Spanish language HRHS (SHRHS). The second study back-translated the SHRHS, which demonstrated a strong correlation with the first translation obtained by Boytell, Velasco-Whetsell, and Coffin, further supporting the accuracy and reliability of the SHRHS.

Neuroendocrine-induced synthesis of bone marrow-derived cytokines with inflammatory immunomodulating properties.

Although cytokines and other soluble regulators of immunity are known to be involved in hematopoiesis, little is known about the signals that induce the synthesis of those mediators locally. Based on recent studies linking the neuroendocrine hormone thyrotropin [thyroid-stimulating hormone (TSH)] to immune cell function in other tissues, we investigated the capacity of TSH to activate cytokine responses from bone marrow cells. These studies reveal that stimulation of the TSH receptor on bone marrow cells-using highly purified or recombinant TSH or by direct stimulation with anti-TSH receptor antibodies-rapidly induces the synthesis of cytokines from bone marrow cells that are classically used in the regulation of inflammatory responses. Of 13 cytokines screened for activity by ELISA or by RNase protection assays for gene expression, IL-6, IFN-beta, TNFalpha, TNFbeta, TGFbeta2, and lymphotoxin-beta responses were reproducibly induced by TSH within 2-3 h of stimulation. Intracellularly, TSH stimulation of bone marrow cells caused rapid increases in cAMP levels and induced the phosphorylation of the Jak2 protein kinase, thereby defining a novel G-protein-coupled receptor/cytokine synthesis pathway. These findings demonstrate that TSH can serve as a primary inductive signal of cytokine production by bone marrow cells.

Children's fears.

The purpose of this article is to provide nurses with a comprehensive literature review of children's fears and to offer interventions that help children cope with fear. Children's normal fear development and the importance of developmental considerations in the assessment, diagnosis, and treatment of fear are also explored. A preventive model based on the work of Robinson and colleagues highlights several approaches to assist children in dealing with fear. This model provides consultation and counseling strategies that the nurse may use to help children develop control, self-worth, and security. This model may also be used to explore areas of normative developmental fears.

T cell progenitors in the murine small intestine.

Lymphocytes in the murine small intestine epithelium are known to have a high proportion of extrathymic T cells. To explore the possibility that small intestine intraepithelial lymphocytes (IELs) are derived from T cell progenitors present within the intestine, intestine-derived cells with characteristics of early-stage T cell precursors were studied for their ability to regenerate IEL T cell populations following transfer into irradiated recipient mice. Cells within this population lacked markers of mature T cells but expressed heat-stable antigen, the c-kit receptor for stem cell factor, and/or the pre-T cell alpha gene. Upon adoptive transfer, donor cells preferentially homed to the intestine and did not repopulate the thymus or extraintestinal peripheral lymphoid tissues. IELs derived from the donor precursor pool included both (alpha beta and gamma delta T subsets and consisted of phenotypically heterogeneous cell populations defined by CD4 and CD8. These findings provide evidence that T cell progenitors located in the intestinal mucosa are the likely source of most intestinal IELs.

Local hormone networks and intestinal T cell homeostasis.

Neuroendocrine hormones of the hypothalamus-pituitary-thyroid axis can exert positive or negative immunoregulatory effects on intestinal lymphocytes. Small intestine epithelial cells were found to express receptors for thyrotropin-releasing hormone (TRH) and to be a primary source of intestine-derived thyroid-stimulating hormone (TSH). The gene for the TSH receptor (TSH-R) was expressed in intestinal T cells but not in epithelial cells, which suggested a hormone-mediated link between lymphoid and nonhematopoietic components of the intestine. Because mice with congenitally mutant TSH-R (hyt/hyt mice) have a selectively impaired intestinal T cell repertoire, TSH may be a key immunoregulatory mediator in the intestine.

T cell precursors in the spleen give rise to complex T cell repertoires in the thymus and the intestine.

T cell precursors located in peripheral immune tissues have been studied according to the potential to repopulate the thymus and the gut of lethally irradiated mice. T cell repopulation could be achieved with spleen cells from athymic or euthymic mice thoroughly devoid of mature T cells. Repopulation did not occur with lymph node lymphocytes as determined from studies in congenic mice. The kinetics of T cell repopulation differed in the gut and thymus in that donor-derived T cells appeared in the gut by day 7 after cell transfer, and in the thymus by day 14 after cell transfer. The multipotent nature of splenic T cell precursors was evident from the finding that all major phenotypic subsets of T cells in the thymus and the gut developed after spleen cell transfer. T cell repopulation of the intraepithelial lymphocytes also occurred efficiently in athymic radiation chimeras injected with spleen cells from congenitally athymic nude mice, demonstrating that gut T cell repopulation by those cells does not require a functional thymus. PCR-spectratype analyses of twenty-four V beta TCR genes in thymocytes and intraepithelial lymphocytes revealed extensive TCR-beta repertoires in both tissues 1 to 2 wk after cell transfer, although T cells with rearranged TCR were undetectable in the donor spleen cell population. The minimal phenotype of the splenic T cell precursor was determined to be CD3-, CD4-, CD8-, HSA+.

A novel marker of murine bone marrow hematopoietic stem cells that is expressed on peripheral T cells and is associated with a functionally important molecule on activated cytotoxic T lymphocytes.

A MAb (R2/60) has been isolated that defines a novel lymphocyte marker of murine T cells. The determinant recognized by MAb R2/60 is present on a subset of bone marrow (BM) hematopoietic stem cells, on adult thymocytes, on peripheral T cells (both resting and activated), and on murine T cell tumor lines, although it is not expressed on mature B cells. In immunoprecipitation studies using radiolabeled membrane lysates from adult thymocytes, MAb R2/60 precipitated a 44-kDa membrane-bound dimer. Functionally, MAb R2/60 mediated antigen-independent cell lysis by activated CTLs, and by CTL clones, when bridged to Fc receptor-bearing target cells; however, binding of MAb R2/60 to effector cells prior to cytotoxic assays did not inhibit target cell lysis by CTLs, suggesting that the R2/60 determinant is involved in transmembrane signaling to already activated CTLs, but that it is not involved in target cell adhesion or antigen recognition. Moreover, direct stimulation of T cells by MAb R2/60 in the absence of additional stimuli did not induce cell proliferation, further implying that the R2/60 determinant is functionally involved in the effector rather than the inductive phase of the T cell response.

Phenotype and TCR gamma delta variable gene repertoire of intestinal intraepithelial lymphocytes in wild mice (Mus musculus domesticus): abundance of V gamma 1 transcripts and extensive delta gene diversity.

In order to study murine intestinal intraepithelial lymphocytes (IEL) independent of factors imparted by conditions of laboratory housing and breeding, and to provide a basis for comparison of IEL studies between inbred and outbred mouse populations, IEL from the domestic house mouse, Mus musculus domesticus, were analyzed by flow cytometric analyses using mAbs to murine lymphocyte markers, and by polymerase chain reaction to study the TCR gamma and delta V gene repertoires. The majority of IEL in wild mice were CD3+, CD8+CD4- T cells. CD4+CD8- also were present in IEL isolates from wild mice, although at low numbers. Among IEL, but not T cells from the spleen or lymph nodes, there was a notable lack of Thy-1 expression, a preponderance of CD8 alpha alpha + T cells, and a relatively high ratio (3:1) of TCR gamma delta + T cells over TCR alpha beta + T cells, suggesting that some IEL in wild mice may develop via an extrathymic pathway similar to that described for laboratory mice. Analyses of the IEL gamma and delta variable genes revealed rearrangements of three of six V region gamma genes (V gamma 1, V gamma 2, and V gamma 5), with an abundance of V gamma 1 transcripts as determined by Northern blot analyses. For the delta gene, rearrangement of five of seven V region elements had occurred (V delta 2, V delta 3, V delta 4, V delta 5, and V delta 6).(ABSTRACT TRUNCATED AT 250 WORDS)

T cell receptor delta gene repertoire and diversity of intestinal intraepithelial lymphocytes in athymic mice.

T cell receptor (TCR) delta gene rearrangements in intestinal intraepithelial lymphocytes (IEL) were studied in athymic radiation chimeras using polymerase chain reaction (PCR) and sequence analysis of DNAs spanning the variable (V), diversity (D), and junctional (J) genes. In both thymus-bearing and athymic mice, IEL delta gene rearrangements occurred for V delta 3, V delta 4, V delta 5 and V delta 6. V-D-J junctional-site sequence analyses of cloned DNAs from rearranged IEL delta genes in athymic mice revealed a predominance of in-frame rearrangements; junctional diversity consisting of nucleotide removal from V, D and/or J genes; N segment nucleotide insertions; and high overall gene diversity. Evaluation of PCR-amplified cDNAs made from IEL RNA indicated that all four rearranged V delta genes were expressed in IEL from athymic mice. The high diversity observed at the gene level also was present in amino acid sequences encoded by the V-D-J region of IEL delta genes in athymic mice. These data demonstrate that there is extensive diversity of rearranged delta genes in IEL which develop extrathymically, and suggest that the delta chain of IEL TCR-gamma delta+ T cells has the potential for interactions with polymorphic structures.

Do postsuctioning transcutaneous PO2 values change when a neonate's movements are restrained?

The purpose of this research was to examine the effects of containment (restraint of an infant's movements) on premature infants' postsuctioning transcutaneous PO2 values. The hypothesis was that premature infants receiving containment would have significantly different postsuctioning transcutaneous PO2 levels than infants receiving no containment. Premature infants with respiratory disease require suctioning to remove excess secretions from their lungs. Research studies document a variety of infant responses to suctioning; hypoxia, hypoxemia, increased blood pressure, bradycardia, increased intracranial pressure, and increased cerebral blood flow velocity. Most studies have examined only a few isolated variables and the magnitude and duration of hypoxemia in response to suctioning. Our sample comprised 24 infants less than 72 hours of age, who had respiratory distress syndrome and had been born at between 24 and 33 weeks' gestational age. The setting was a neonatal intensive care unit of a large county hospital. The same nurse performed the suctioning and containment procedures in all subjects. Student's t-tests and analysis of variance were carried out to determine the effect of the technique.

Rearrangement and junctional-site sequence analyses of T-cell receptor gamma genes in intestinal intraepithelial lymphocytes from murine athymic chimeras.

The molecular organization of rearranged T-cell receptor (TCR) gamma genes intraepithelial lymphocytes (IEL) was studied in athymic radiation chimeras and was compared with the organization of gamma gene rearrangements in IEL from thymus-bearing animals by polymerase chain reaction and by sequence analyses of DNA spanning the junction of the variable (V) and joining (J) genes. In both thymus-bearing mice and athymic chimeras, IEL V-J gamma-gene rearrangements occurred for V gamma 1.2, V gamma 2, and V gamma 5 but not for V gamma 3 or V gamma 4. Sequence analyses of cloned V-J polymerase chain reaction-amplified products indicated that in both thymus-bearing mice and athymic chimeras, rearrangement of V gamma 1.2 and V gamma 5 resulted in in-frame as well as out-of-frame genes, whereas nearly all V gamma 2 rearrangements were out of frame from either type of animal. V-segment nucleotide removal occurred in most V gamma 1.2, V gamma 2, and V gamma 5 rearrangements; J-segment nucleotide removal was common in V gamma 1.2 but not in V gamma 2 or V gamma 5 rearrangements. N-segment nucleotide insertions were present in V gamma 1.2, V gamma 2, and V gamma 5 IEL rearrangements in both thymus-bearing mice and athymic chimeras, resulting in a predominant in-frame sequence for V gamma 5 and a predominant out-of-frame sequence for V gamma 2 genes. These findings demonstrate that (i) TCR gamma-gene rearrangement occurs extrathymically in IEL, (ii) rearrangements of TCR gamma genes involve the same V gene regardless of thymus influence; and (iii) the thymus does not determine the degree to which functional or nonfunctional rearrangements occur in IEL.

Proliferative properties of murine intestinal intraepithelial lymphocytes (IEL): IEL expressing TCR alpha beta or TCR tau delta are largely unresponsive to proliferative signals mediated via conventional stimulation of the CD3-TCR complex.

Murine intestinal intraepithelial lymphocytes (IEL) were studied for their capacity to proliferate in vitro following stimulation of the T cell receptor (TCR)-associated CD3 epsilon molecule, or upon direct stimulation of the TCR complex itself. Although IEL consisted primarily of CD3+ T cells which included activated cytotoxic T lymphocytes as demonstrated in CD3- and TCR-mediated redirected cytotoxic assays, IEL displayed minimal proliferative responses following stimulation with anti-CD3, anti-TCR alpha beta, or anti-TCR tau delta monoclonal antibodies under soluble conditions, or under conditions which effect membrane cross-linking, including the addition of accessory cells to IEL cultures. The lack of proliferation induction could not be overcome by stimulation of IEL in the presence of T cell-dependent cytokines, phorbol ester, or interleukin-4. Moreover, unlike splenic T cells, stimulation of IEL failed to result in expression of interleukin-2 receptor, further demonstrating an inability of IEL to respond to exogenous proliferative signals. This study is the first to examine the proliferative potential of murine IEL following direct CD3 or TCR stimulation. The findings described here: (i) identify an important functional distinction between intestinal IEL and other peripheral alpha beta or tau delta T cells which generally respond well to proliferative signals mediated through the CD3-TCR complex, and (ii) demonstrate that on murine IEL the CD3-TCR complex can discriminate signals of lytic activity from those of cell proliferation.