Partial budget analysis of culture- and algorithm-guided selective dry cow therapy.

The objectives of this study were to (1) use partial budget analysis to estimate the cash impact for herds that switch from blanket dry cow therapy (BDCT) to culture- or algorithm-guided selective dry cow therapy (SDCT) and (2) conduct a sensitivity analysis to investigate effects in situations where SDCT increased clinical and subclinical mastitis risk during the subsequent lactation. A partial budget model was created using Monte Carlo simulation with @Risk software. Expenditures associated with dry-off procedures and health outcomes (clinical and subclinical mastitis) during the first 30 d in milk were used to model herd-level effects, expressed in units of US dollars per cow dry-off. Values for each economic component were derived from findings from a recent multisite clinical trial, peer-reviewed journal articles, USDA databases, and our experiences in facilitating the implementation of SDCT on farms. Fixed values were used for variables expected to have minimal variation within the US dairy herd population (e.g., cost of rapid culture plates) and sampling distributions were used for variables that were hypothesized to vary enough to effect the herd net cash impact of one or more DCT approach(es). For Objective 1, herd-level udder health was assumed to be unaffected by the implementation of SDCT. For culture-guided SDCT, producers could expect to save an average of +$2.14 (-$2.31 to $7.23 for 5th and 95th percentiles) per cow dry-off as compared with BDCT, with 75.5% of iterations being ≥$0.00. For algorithm-guided SDCT, the mean net cash impact was +$7.85 ($3.39-12.90) per cow dry-off, with 100% of iterations being ≥$0.00. The major contributors to variance in cash impact for both SDCT approaches were percent of quarters treated at dry-off and the cost of dry cow antibiotics. For Objective 2, we repeated the partial budget model with the 30-d clinical and subclinical mastitis incidence increasing by 1, 2, and 5% (i.e., risk difference = 0.01, 0.02, and 0.05) in both SDCT groups compared with BDCT. For algorithm-guided SDCT, average net cash impacts were ≥$0.00 per cow dry-off (i.e., cost effective) when mastitis incidence increased slightly. However, as clinical mastitis incidence increased, economic returns for SDCT diminished. These findings indicate that when SDCT is implemented appropriately (i.e., no to little negative effect on health), it might be a cost-effective practice for US herds under a range of economic conditions.

Development of a novel antibody to calcitonin gene-related peptide for the treatment of osteoarthritis-related pain.

OBJECTIVE: Investigate a role for calcitonin gene-related peptide (CGRP) in osteoarthritis (OA)-related pain. DESIGN: Neutralizing antibodies to CGRP were generated de novo. One of these antibodies, LY2951742, was characterized in vitro and tested in pre-clinical in vivo models of OA pain. RESULTS: LY2951742 exhibited high affinity to both human and rat CGRP (KD of 31 and 246 pM, respectively). The antibody neutralized CGRP-mediated induction of cAMP in SK-N-MC cells in vitro and capsaicin-induced dermal blood flow in the rat. Neutralization of CGRP significantly reduced pain behavior as measured by weight bearing differential in the rat monoiodoacetate model of OA pain in a dose-dependent manner. Moreover, pain reduction with neutralization of CGRP occurred independently of prostaglandins, since LY2951742 and NSAIDs worked additively in the NSAID-responsive version of the model and CGRP neutralization remained effective in the NSAID non-responsive version of the model. Neutralization of CGRP also provided dose-dependent and prolonged (>60 days) pain reduction in the rat meniscal tear model of OA after only a single injection of LY2951742. CONCLUSIONS: LY2951742 is a high affinity, neutralizing antibody to CGRP. Neutralization of CGRP is efficacious in several OA pain models and works independently of NSAID mechanisms of action. LY2951742 holds promise for the treatment of pain in OA patients.

Increased myocardial Rab GTPase expression: a consequence and cause of cardiomyopathy.

The Ras-like Rab GTPases regulate vesicle transport in endocytosis and exocytosis. We found that cardiac Rabs1, 4, and 6 are upregulated in a dilated cardiomyopathy model overexpressing beta(2)-adrenergic receptors. To determine if increased Rab GTPase expression can contribute to cardiomyopathy, we transgenically overexpressed in mouse hearts prototypical Rab1a, the small G protein that regulates vesicle transport from endoplasmic reticulum to and through Golgi. In multiple independent mouse lines, Rab1a overexpression caused cardiac hypertrophy that progressed in a time- and transgene dose-dependent manner to heart failure. Isolated cardiac myocytes were hypertrophied and exhibited contractile depression with impaired calcium reuptake. Ultrastructural analysis revealed enlarged Golgi stacks and increased transitional vesicles in ventricular myocytes, with increased secretory atrial natriuretic peptide granules and degenerative myelin figures in atrial myocytes; immunogold studies localized Rab1a to these abnormal vesicular structures. A survey of hypertrophy signaling molecules revealed increased protein kinase C (PKC) alpha and delta, and confocal microscopy showed abnormal subcellular distribution of PKCalpha in Rab1a transgenics. These results indicate that increased expression of Rab1 GTPase in myocardium distorts subcellular localization of proteins and is sufficient to cause cardiac hypertrophy and failure.

Lead-contaminated imported tamarind candy and children's blood lead levels.

In 1999, an investigation implicated tamarind candy as the potential source of lead exposure for a child with a significantly elevated blood lead level (BLL). The Oklahoma City-County Health Department tested two types of tamarind suckers and their packaging for lead content. More than 50% of the tested suckers exceeded the US Food and Drug Administration (FDA) Level of Concern for lead in this type of product. The authors calculated that a child consuming one-quarter to one-half of either of the two types of suckers in a day would exceed the maximum FDA Provis onal Tolerable Intake for lead. High lead concentrations in the two types of wrappers suggested leaching as a potential source of contamination. The authors used the Environmental Protection Agency's Integrated Exposure Uptake Biokinetic (IEUBK) model to predict the effects of consumption of contaminated tamarind suckers on populat on BLLs. The IEUBK model predicted that consumption of either type of sucker at a rate of one per day would result in dramatic increases in mean BLLs for children ages 6-84 months in Oklahoma and in the percentage of children wth elevated BLLs (> or =10 micrograms per deciliter [microg/dL]). The authors conclude that consumption of these products represents a potential public health threat. In addition, a history of lead contamination in imported tamarind products suggests that import control measures may not be completely effective in preventing additional lead exposure.

hREC2, a RAD51-like gene, is disrupted by t(12;14) (q15;q24.1) in a uterine leiomyoma.

A balanced translocation between chromosomes 12 and 14 is commonly seen in uterine leiomyoma (UL). We have previously cloned and characterized a 2 Mb segment of human chromosomal subband 14q24.1, and have shown that the t(12;14)(q15;q24.1) breakpoints from several ULs map within this region. Exon trapping of DNA clones spanning one such breakpoint revealed coding sequences from hREC2, a gene that shows significant amino acid sequence identity to the double-strand break repair enzyme RAD51. We report here that this breakpoint is located within a 19 kb intron of the hREC2 gene and that the translocation results in the premature truncation of the major hREC2 transcript. Mapping and sequence analyses show that alternative transcripts of the hREC2 gene, including novel isoforms identified in testis and uterus, are not interrupted by the translocation.

Sensitive RIA for the specific determination of insulin lispro.

Insulin lispro is an insulin analog in which the primary sequence has been altered by the inversion of amino acids B28 and B29. To date, it has not been possible to specifically measure insulin lispro in the presence of endogenous insulin because of the high degree of homology between these peptides. However, the specific determination of insulin lispro offers advantages over quantifying total concentrations of immunoreactive insulin. We therefore immunized guinea pigs and screened for antibodies with increased affinity and selectivity for insulin lispro. We prepared a monospecific antiserum by a novel immunoadsorption strategy using despentapeptide insulin. The antiserum was used to develop a competitive RIA for insulin lispro. The RIA has a low limit of quantification (17.2 pmol/L); has no interference from insulin, proinsulin, or C-peptide; and has interassay CVs of 2.6-13.4%. The new RIA is useful for measuring serum concentrations of insulin lispro.

Genomic and functional map of the chromosome 14 t(12;14) breakpoint cluster region in uterine leiomyoma.

A translocation involving chromosomes 12 and 14 [t(12;14)(q15;24.1)] is commonly seen in benign smooth muscle tumor as uterine leiomyoma (UL). A contig of P1-derived artificial chromosome and bacterial artificial chromosome clones on chromosome 14, encompassing a t(12;14) breakpoint cluster region (BCR) in UL, was generated principally using the recently developed HAPPY map of chromosome 14 as a framework (P. H. Dear et al., 1998, Genomics 48: 232-241). Three UL t(12;14) breakpoints have been localized within this contig, showing that a BCR of at least 400 kb exists on chromosome 14. Other studies of tumors with t(12;14) rearrangements similarly show breakpoints within a 475-kb multiple aberration region on chromosome 12. Thus t(12;14) is an example of a translocation in which the breakpoints are located within a BCR on both chromosome 12 and chromosome 14, justifying the identification of expressed sequences that are altered in these BCR regions. A total of four expressed sequences were identified in the BCR on chromosome 14. Two of these were novel cDNAs (D14S1460E and D14S1461E). The chromosome 14 cDNAs were expressed in multiple adult tissues. The identification of a large breakpoint cluster region on chromosome 14 suggests that translocations in this region mediate their effects at a distance and also that elements that predispose this region to recurrent chromosomal translocation may be widely distributed.

Cloning of a breakpoint cluster region on chromosome 14 in uterine leiomyoma.

A recurrent reciprocal chromosomal translocation, t(12;14)(q15;q24) is frequently observed in uterine leiomyoma. Chromosome 12 breakpoints have been shown to occur in a region of approximately 150 kb that contains the gene for a high mobility group protein (HMGI-C). The breakpoint region on chromosome 14 has not been precisely defined. We have generated a contig of overlapping yeast artificial chromosome (YAC) clones approximately 3 Mb in size. Fluorescence in situ hybridization (FISH) analysis showed that this contig spanned the t(12;14) breakpoints in three uterine leiomyomas and that the breakpoints in these tumors occurred within a 1 Mb region. A 30 kb cosmid spanning one of the breakpoints was isolated to set the stage for identifying regions on chromosome 14 that may cause this region to be a preferential site for chromosomal translocation.

Pyomyositis of the anterior tibial compartment.

Five oncology patients developed bacterial pyomyositis involving the anterior tibial compartment and resulting in compartment syndrome with ischemia and abnormalities of neuromuscular function. All patients were neutropenic and thrombocytopenic, and four were receiving or had recently received cancer chemotherapy. Three infections were due to gram-negative bacilli and two to Staphylococcus aureus. Appropriate antimicrobial therapy and surgical drainage in four patients resulted in the resolution of these infections with good residual muscle function. To our knowledge, primary pyomyositis has never previously been known to cause compartment syndrome.

Expression of N-cadherin and alpha-catenin in astrocytomas and glioblastomas.

We examined levels of mRNA and protein for N-cadherin, the predominant cadherin in neural tissues, and mRNA levels for the cadherin-associated protein, alpha-catenin, in a series of gliomas and in glioblastoma cell lines. mRNA levels for N-cadherin and alpha-catenin were significantly higher in glioblastomas than in low-grade astrocytomas or normal brain, while the levels of intact N-cadherin protein were similar in glioblastomas, low-grade astrocytomas and brain. In addition, there was no consistent relationship between invasiveness and expression of N-cadherin and alpha-catenin in highly invasive vs minimally invasive tumours within the same histopathological grade. To assess further the relationship between cadherin expression and neural tumour invasion, we measured N-cadherin expression, calcium-dependent cell adhesion and motility of several glioblastoma cell lines. While all N-cadherin-expressing lines were adhesive, no correlation was seen between the level of N-cadherin expression and cell motility. Together, these findings imply that, in contrast to the role played by E-cadherin in carcinomas, N-cadherin does not restrict the invasion of glioblastomas.

Managing geriatric arrhythmias, II: Drug selection and use.

Age-related impairments in antiarrhythmic drug distribution, metabolism, and excretion may result in accumulation of these potent drugs, increasing the risk of adverse drug reactions and drug interactions in the elderly. These risks are minimized by using modest initial dosage with slow dosage titration. Therapeutic drug monitoring is a method which employs antiarrhythmic concentration data and a determination of drug clearance in order to develop an optimal dosing regimen. The pharmacology, indications for use, and adverse effects of currently available antiarrhythmics are reviewed. Dosing guidelines for the geriatric population are provided.

Managing geriatric arrhythmias, I: General considerations.

Cardiac arrhythmias become increasingly common as people age, but they are not always clinically significant. Sinus node dysfunction, AV conduction disturbances, and ventricular and supraventricular arrhythmias will be found but are not always symptomatic. Treatment of asymptomatic arrhythmias is controversial and probably not indicated, but in symptomatic elderly, therapy is indicated, since even the very elderly have been found to benefit from it as much as younger patients. There are specific guidelines, however, that apply to this age group based on its susceptibility to side effects.

Poly-N-acetyllactosamine alterations of Thy-1 glycoprotein in lymphocyte differentiation.

Developmental changes in murine lymphocyte Thy-1 include both quantitative and qualitative alterations involving N-linked oligosaccharides. Comparison of immature with mature T-cells has shown that the oligosaccharides of Thy-1 are characterized by an increase in the number of sialic acid residues responsible for the acidic pI of peripheral T-cell Thy-1, and a decrease in those oligosaccharides responsible for Mr heterogeneity of thymocyte Thy-1. The research reported here suggests that the basis of the large Mr heterogeneity in Thy-1 of immature T-cells is the presence of repeating N-acetyllactosamine (R'Gal beta 1,4GlcNAc beta 1,3R") units in the oligosaccharide portion of the molecule. Lymphocytes were surface iodinated and 125I-thy-1 was purified by immunoprecipitation and preparative nonequilibrium pH gradient electrophoresis. The minimal Mr of unglycosylated Thy-1 after endoglycosidase F digestion was 15,000-16,000. Digestion of Thy-1 with endo-beta-galactosidase suggested that the complex type N-linked glycans in thymocytes, but not in lymph node T-cells, contained increased levels of polylactosamine. The presence of polylactosamine was confirmed by binding to a Datura stramonium lectin column which retarded and bound approx. 50% of thymocyte Thy-1 and only about 18% of lymph node T-cell Thy-1. Affinity chromatography using anti-i antibody immobilized on agarose beads indicated that the polylactosamine is probably present in a predominantly linear form. Since alterations of polylactosamine structures have been implicated in development and transformation in several systems, the present results suggest an important role for these glycans in immune-cell differentiation.