Genetic change results from selection on an economic breeding objective in beef cattle.

The objective of this study was to evaluate and quantify the genetic progress achieved in a New Zealand Angus nucleus herd through long-term selection for an economically based, multi-trait breeding objective. A 4-trait breeding objective was implemented in 1976 and selected on through 1993 with traits consisting of slaughter weight and dressing percentage of harvest progeny and cull cows, and the number of calves weaned in the lifetime of each cow. These traits were related to gross income with none related to costs of production. To overcome this, economic weights were adjusted down for increased feed requirements of faster growing (and generally larger) animals. Performance and pedigree information was recorded on 16,189 animals from 1976 through 1993 and included weaning, yearling, and mature cow weights along with the lifetime number of calves weaned by each cow. These traits were used in the phenotypic selection indexes developed to predict the defined breeding objective. Individual performance was adjusted by least squares for major environmental fixed effects and deviated from contemporaneous means. Genetic and residual (co)variances were re-estimated for each of the traits using REML techniques and used to calculate EBV for each trait. These EBV were in turn used to calculate annual genetic changes. The average annual genetic changes for weaning weight direct and maternal breeding value were 0.43 +/- 0.05 and 0.03 +/- 0.22 kg/yr, respectively. Corresponding annual genetic changes for postweaning BW gain, yearling weight, harvest weight, and mature BW were 0.29 +/- 0.03, 0.72 +/- 0.06, 1.7 +/- 0.13, and 0.13 +/- 0.09 kg, respectively. The annual change in number of calves weaned per cow lifetime was 0.006 +/- 0.001 calves/cow and the change in dressing percentage was estimated to be -0.035 +/- 0.003 %/yr. At the end of the program, 3.21 generations of selection had occurred with a mean accumulated selection differential of 3.87 SD. Change in objective traits due to selection was similar to or exceeded change predicted at the onset of the program with the exception of mature BW and dressing percentage. Genetic change in mature BW was not different from zero, whereas the predicted change was 29.3 kg. The overall genetic trend in the breeding objective exceeded that predicted at the onset of the program. Results of this study showed that selection on indexes developed to predict an economically based, multi-trait breeding objective will produce genetic change.

Persistent airway hyper-responsiveness and inflammation in Toxocara canis-infected BALB/c mice.

BACKGROUND: Infection with Toxocara canis, the roundworm of dogs, has been associated with asthmatic manifestations. Clinical symptoms such as wheezing, coughing and episodic airflow obstruction have been described for patients infected with this helminth. OBJECTIVE: In order to characterize the effect of T. canis infection on the lungs, we monitored immune responses, pulmonary pathology and lung function over a period of 60 days in BALB/c mice. METHODS: Infection was performed by a single oral administration of 1000 T. canis embryonated eggs. Airway responsiveness was measured in conscious, unrestrained mice at 7, 14, 30 and 60 days post-infection (p.i.). RESULTS: Infection of mice resulted in airway hyper-responsiveness (AHR) that persisted up to 30 days p.i. Pulmonary inflammation as well as increased levels of IgE and eosinophils in bronchoalveolar lavage (BAL) persisted up to 60 days p.i. Cytokine analysis in BAL indicated increased levels of IL-5 at day 7 and 14 p.i., whereas the levels of IL-2, IFN-gamma, IL-4 and IL-10 did not differ from those of uninfected controls. Toxocara-specific stimulation of spleen cells using recombinant TES-70 protein resulted in the induction of IL-5 at day 7 and 14 p.i. and IL-10 at day 14 p.i. Production of all other cytokines did not differ from that of uninfected controls. Evaluation of larval burden revealed that T. canis was still present in the lungs of infected mice at 60 days p.i. CONCLUSION: The presence of Toxocara larva in the lungs at 60 days p.i. following a single infection could explain the persistent pulmonary inflammation, airway hyper-reactivity, eosinophilia and increased IgE production observed in T. canis-infected BALB/c mice.

Expression of the Hypermethylated in Cancer gene (HIC-1) is associated with good outcome in human breast cancer.

A new cancer gene, HIC-1 (Hypermethylated in Cancer) telomeric to p53 on chromosome 17p may be of clinical importance in sporadic breast cancer. Regional DNA hypermethylation of 17p13.3 resulting in suppression of gene expression has been shown to precede 17p structural changes in human carcinogenesis. In addition, loss of heterozygosity studies have suggested clinically significant involvement of a gene on 17p13.3 associated with poor prognosis in breast cancer. Using RT-PCR analysis, we demonstrate that the MCF7 (wild type p53) cell line expressed HIC-1 transcripts but the MDAMB231 (mutant p53) cell line did not, suggesting loss of HIC-1 expression and p53 malfunction may be synergistic events in sporadic breast cancer. HIC-1 expression was examined using RT-PCR on RNA extracted from 50 primary untreated, human breast cancers and was detected in only 7/50 (14%) cancers. All seven patients with HIC-1 expression were alive without disease recurrence after 8 years follow-up and 5/7 had detectable p53 wild type mRNA expression. This suggests that retained HIC-1 expression may offer a survival advantage. However the seven cancers had 17p13.3 loss of heterozygosity (LOH; four patients), a feature previously associated with poor prognosis, or were homozygous (three patients) suggesting there may be two genes at 17p13.3 involved in breast carcinogenesis. Using a demethylating drug 5-aza-2'-deoxycytidine (DeoxyC), HIC-1 expression was restored in the MDAMB231 cells, also suggesting restoration of HIC-1 function by reversing HIC-1 hypermethylation may offer a therapeutic avenue in breast cancer.

Identification, characterization and leucocyte expression of Siglec-10, a novel human sialic acid-binding receptor.

Here we characterize Siglec-10 as a new member of the Siglec family of sialic acid-binding Ig-like lectins. A full-length cDNA was isolated from a human spleen library and the corresponding gene identified. Siglec-10 is predicted to contain five extracellular Ig-like domains and a cytoplasmic tail containing three putative tyrosine-based signalling motifs. Siglec-10 exhibited a high degree of sequence similarity to CD33-related Siglecs and mapped to the same region, on chromosome 19q13.3. The expressed protein was able to mediate sialic acid-dependent binding to human erythrocytes and soluble sialoglycoconjugates. Using specific antibodies, Siglec-10 was detected on subsets of human leucocytes including eosinophils, monocytes and a minor population of natural killer-like cells. The molecular properties and expression pattern suggest that Siglec-10 may function as an inhibitory receptor within the innate immune system.

Siglec-9, a novel sialic acid binding member of the immunoglobulin superfamily expressed broadly on human blood leukocytes.

Here we characterize the properties and expression pattern of Siglec-9 (sialic acid-binding Ig-like lectin-9), a new member of the Siglec subgroup of the immunoglobulin superfamily. A full-length cDNA encoding Siglec-9 was isolated from a dibutyryl cAMP-treated HL-60 cell cDNA library. Siglec-9 is predicted to contain three extracellular immunoglobulin-like domains that comprise an N-terminal V-set domain and two C2-set domains, a transmembrane region and a cytoplasmic tail containing two putative tyrosine-based signaling motifs. Overall, Siglec-9 is approximately 80% identical in amino acid sequence to Siglec-7, suggesting that the genes encoding these two proteins arose relatively recently by gene duplication. Binding assays showed that, similar to Siglec-7, Siglec-9 recognized sialic acid in either the alpha2,3- or alpha2, 6-glycosidic linkage to galactose. Using a specific mAb, Siglec-9 was found to be expressed at high or intermediate levels by monocytes, neutrophils, and a minor population of CD16(+), CD56(-) cells. Weaker expression was observed on approximately 50% of B cells and NK cells and minor subsets of CD8(+) T cells and CD4(+) T cells. These results show that despite their high degree of sequence similarity, Siglec-7 and Siglec-9 have distinct expression profiles.

Identification and characterization of a novel siglec, siglec-7, expressed by human natural killer cells and monocytes.

We describe the characterization of sialic acid-binding Ig-like lectin-7 (siglec-7), a novel member of the siglec subgroup of the immunoglobulin superfamily. A full-length cDNA encoding siglec-7 was isolated from a human primary dendritic cell cDNA library. Siglec-7 is predicted to contain three extracellular immunoglobulin-like domains that comprise an N-terminal V-set domain and two C2-set domains, a transmembrane region and a cytoplasmic tail containing two tyrosine residues embodied in immunoreceptor tyrosine-based inhibition motif-like motifs. Overall, siglec-7 exhibited a high degree of sequence similarity to genes encoding CD33 (siglec-3), siglec-5, OBBP1/siglec-6, and OBBP-like protein and mapped to the same region on chromosome 19q13.3. When siglec-7 was expressed on COS or Chinese hamster ovary cells, it was able to mediate high levels of sialic acid-dependent binding to human erythrocytes and soluble sialoglycoconjugates, suggesting that it may be involved in cell-cell interactions. Among human peripheral blood leukocytes, siglec-7 was found to be present at low levels on granulocytes, intermediate levels on monocytes, and relatively high levels on a major subset of natural killer cells and a minor subset of CD8(+) T cells. Immunoprecipitation experiments indicated that siglec-7 is expressed as a monomer of approximately 65 kDa.

The irritancy of soaps and syndets.

The use of soaps and synthetic detergent (syndet) bars has been associated with skin dryness and aggravation of dermatologic conditions. A number of factors, including chemical structure, pH, and cleansing ability, have been implicated in this phenomenon. Many bars contain agents designed to ameliorate the irritancy of the bar and/or provide a skin benefit. Clinical studies have shown that soaps are generally quite irritating while syndets can range from mild to harsh. The addition of skin benefit/mildness agents such as glycerin, cocoa butter, mineral oil, or lanolin has little effect on the irritancy potential of a bar since minimal amounts of these agents are deposited on the skin. The excessive removal of skin lipids by harsh soaps and detergents can result in superficial dryness. The key to gentle cleansing is to start with a mild cleansing product and avoid overuse.

Muscle and liver pyruvate kinases are closely related: amino acid sequence comparisons.

Previous evidence has shown that the M1 and L pyruvate kinase isozymes differ markedly in kinetic and immunological properties, amino acid compositions and peptide maps. However, the amino acid sequence results we present here for the N-terminal region and for a region of the C domain show that the M1 and L isozymes are very similar. The variable length of the N-terminal sequences also explains the difference in regulation by phosphorylation between the M1 and L isozymes. The M1 isozyme lacks the serine residue that has been shown to be phosphorylated in the L isozyme.

Quantitation of macrophage activation.

Chemoluminescence is a sensitive and relatively easy method to quantitate macrophage activation. Cytotoxic assay as measured by 51Cr release from tumour target cells is more involved and less sensitive. Our experiments indicate that, depending on the mode of activation, different effector mechanisms are operational. Thus, macrophages activated with double stranded yeast RNA show good chemoluminescence but no cytotoxic response while thioglycollate activated macrophages show the reverse response.

In vivo crevicular leukocyte response to a chemotactic challenge: inhibition by experimental diabetes.

Diabetes in rats inhibits the migration of neutrophils into the healing gingival crevice, an effect associated with impaired in vitro neutrophil chemotactic activity. We recently described the in vivo response of human and rat crevicular neutrophils to a chemotactic challenge and used this assay in the present study on streptozotocin-induced diabetic rats. Optimal concentrations of two chemotactic agents, casein (0.2 mul, 2 mg/ml) or N-formylmethionylleucylphenylalanine (0.2 mul, 10(-4) M), were placed into the gingival crevices of control and diabetic rats (time zero) after the resting neutrophil count was measured. After a 15-min delay, the neutrophil counts and gingival crevicular fluid flow were assessed every 5 min for another 0.5 h. The control rats (n = 14) showed an increase in neutrophil counts which reached maximum levels 30 min after the N-formylmethionylleucylphenylalanine challenge ("peak" neutrophil response) and decreased dramatically 5 min later. Diabetes of 4 days (n = 4), 14 days (n = 8), and 20 days (n = 5) duration reduced the peak neutrophil response 45, 66, and 71%, respectively. Casein produced the same response as N-formylmethionylleucylphenylalanine in control rats. Uncontrolled diabetes of 20 days duration reduced the peak neutrophil response to casein by 83%; diabetics administered insulin on a daily basis showed a reduction of only 34%. The pattern of change in gingival crevicular fluid flow in response to chemoattractants paralleled the neutrophil response. The chemotactic activity of peritoneal neutrophils was assessed in vitro with the agarose gel technique and was found to be correlated (r = 0.84; P < 0.01) with the in vivo chemotactic response in the same rats. If the same in vivo defect is observed in humans with diabetes (or with other systemic diseases associated with leukocyte dysfunction), this test could be useful diagnostically to rapidly assess neutrophil chemotaxis in lieu of in vitro assays and to identify patients who are unusually susceptible to aggressive periodontal disease.

Suprapubic prostatectomy. The removable purse string: a continuing comparative analysis of 300 consecutive cases.

Mortality rates, length of hospitalization, transfusion requirements, weights of excised tissue, complications and operating time are analyzed in this series of 300 patients who underwent suprapubic prostatectomies in which a removable purse-string closure of the bladder neck was used. The results are compared to those in other studies. The mortality rate in our study is the lowest reported to date for a large series. We believe that this technically simple operation, which has the most adaptability for combination with conjunctive operations, low transfusion requirements, few long-term complications and which can be done rapidly, is the most desirable method to remove benign prostates of 40 gm. or larger. We believe that there should be acceptance of broader indications for its use and that greater emphasis should be placed on teaching this procedure in urological training programs.