Cytokine targeting in tumors using a bispecific antibody directed against carcinoembryonic antigen and tumor necrosis factor alpha.

The use of tumor necrosis factor alpha (TNFalpha) in cancer therapy is limited by its short circulatory half-life and its severe systemic side effects. To overcome these limitations, we evaluated the capability of a bispecific antibody (BAb) directed against carcinoembryonic antigen (CEA) and human TNFalpha to target this cytokine in tumors. A BAb was constructed by coupling the Fab' fragments from an anti-CEA monoclonal antibody (MAb) to the Fab' fragments from an anti-TNFalpha MAb via a stable thioether linkage. The double specificity of the BAb for CEA and TNFalpha was demonstrated using a BIAcoreTM two-step analysis. The affinity constants of the BAb for CEA immobilized on a sensor chip and for soluble TNFalpha added to the CEA-BAb complex were as high as those of the parental MAbs (1.7 x 10(9) M-1 and 6.6 x 10(8) M-1, respectively). The radiolabeled 125I-labeled BAb retained high immunoreactivity with both CEA and TNFalpha immobilized on a solid phase. In nude mice xenografted with the human colorectal carcinoma T380, the 125I-labeled BAb showed a tumor localization and biodistribution comparable to that of 131I-labeled anti-CEA parental F(ab')2 with 25-30% of the injected dose (ID)/g tumor at 24 h and 20% ID/g tumor at 48 h. To target TNFalpha to the tumor, a two-step i.v. injection protocol was used first, in which a variable dose of 125I-labeled BAb was injected, followed 24 or 48 h later by a constant dose of 131I-labeled TNFalpha (1 microg). Mice pretreated with 3 microg of BAb and sacrificed 2, 4, 6, or 8 h after the injection of TNFalpha showed a 1.5- to 2-fold increased concentration of 131I-labeled TNFalpha in the tumor as compared to control mice, which received TNFalpha alone. With a higher dose of BAb (25 microg), mice showed a better targeting of TNFalpha with a 3.2-fold increased concentration of 131I-labeled TNFalpha in the tumor: 9.3% versus 2.9% ID/g in control mice 6 h after TNFa injection. In a one-step injection protocol using a premixed BAb-TNFalpha preparation, similar results were obtained 6 h postinjection (3.5-fold increased TNFalpha tumor concentration). A longer retention time of TNFalpha was observed leading to an 8.1-fold increased concentration of TNFalpha in the tumor 14 h postinjection (4.4 versus 0.5% ID/g tumor for BAb-treated and control mice, respectively). These results show that our BAb is able, first, to localize in a human colon carcinoma and, there, to immunoabsorb the i.v.-injected TNFalpha, leading to its increased concentration at the tumor site.

The scaffold protein IB1/JIP-1 controls the activation of JNK in rat stressed urothelium.

The c-Jun N-terminal kinase (JNK) is critical for cell survival, differentiation, apoptosis and tumorigenesis. This signalling pathway requires the presence of the scaffold protein Islet-Brain1/c-Jun N-terminal kinase interacting protein-1 (IB1/JIP-1). Immunolabeling and in situ hybridisation of bladder sections showed that IB1/JIP-1 is expressed in urothelial cells. The functional role of IB1/JIP-1 in the urothelium was therefore studied in vivo in a model of complete rat bladder outlet obstruction. This parietal stress, which is due to urine retention, reduced the content of IB1/JIP-1 in urothelial cells and consequently induced a drastic increase in JNK activity and AP-1 binding activity. Using a viral gene transfer approach, the stress-induced activation of JNK was prevented by overexpressing IB1/JIP-1. Conversely, the JNK activity was increased in urothelial cells where the IB1/JIP-1 content was experimentally reduced using an antisense RNA strategy. Furthermore, JNK activation was found to be increased in non-stressed urothelial cells of heterozygous mice carrying a selective disruption of the IB1/JIP-1 gene. These data established that mechanical stress in urothelial cells in vivo induces a robust JNK activation as a consequence of regulated expression of the scaffold protein IB1/JIP-1. This result highlights a critical role for that scaffold protein in the homeostasis of the urothelium and unravels a new potential target to regulate the JNK pathway in this tissue.

Affirmative Action in Iowa, September 2005

In accordance with 19B.5 of the Code of Iowa, the 2005 Affirmative Action in Iowa report illustrates the progress made during fiscal year 2005 to balance the State's worforce, the challenges that the State must address and the effort that the Department of Administratie Services must lead in order to remove barriers that limit the hiring, retention and advancement of females, minorities and persons with disabilities in the State's workforce. Highlighted in the report are four departments that initiated proactive and innovative measures to address their workplace equal opportunity, affirmative action and diversity programs. Additionally, the Department of Administrative Services-Human Resources Enterprise outlines its plan to build on its past efforts as well as pursue new initiatives to partner with advocacy groups and reach out to the commuity more directely to enhance employment opportunities for females, minorities and persons with disabilities in State of Iowa employment.

Renal and neurohormonal responses to increasing levels of lower body negative pressure in men.

BACKGROUND: The stimulation of efferent renal sympathetic nerve activity induces sequential changes in renin secretion, sodium excretion, and renal hemodynamics that are proportional to the magnitude of the stimulation of sympathetic nerves. This study in men investigated the sequence of the changes in proximal and distal renal sodium handling, renal and systemic hemodynamics, as well as the hormonal profile occurring during a sustained activation of the sympathetic nervous system induced by various levels of lower body negative pressure (LBNP). METHODS: Ten healthy subjects were submitted to three levels of LBNP ranging between 0 and -22.5 mm Hg for one hour according to a triple crossover design, with a minimum of five days between each level of LBNP. Systemic and renal hemodynamics, renal water and sodium handling (using the endogenous lithium clearance technique), and the neurohormonal profile were measured before, during, and after LBNP. RESULTS: LBNP (0 to -22.5 mm Hg) induced an important hormonal response characterized by a significant stimulation of the sympathetic nervous system and gradual activations of the vasopressin and the renin-angiotensin systems. LBNP also gradually reduced water excretion and increased urinary osmolality. A significant decrease in sodium excretion was apparent only at -22.5 mm Hg. It was independent of any change in the glomerular filtration rate and was mediated essentially by an increased sodium reabsorption in the proximal tubule (a significant decrease in lithium clearance, P < 0.05). No significant change in renal hemodynamics was found at the tested levels of LBNP. As observed experimentally, there appeared to be a clear sequence of responses to LBNP, the neurohormonal response occurring before the changes in water and sodium excretion, these latter preceding any change in renal hemodynamics. CONCLUSIONS: These data show that the renal sodium retention developing during LBNP, and thus sympathetic nervous stimulation, is due mainly to an increase in sodium reabsorption by the proximal segments of the nephron. Our results in humans also confirm that, depending on its magnitude, LBNP leads to a step-by-step activation of neurohormonal, renal tubular, and renal hemodynamic responses.

Dynamics of diversity within the Mossos d���Esquadra

This chapter offers a case-study of diversity and diversity policies withinthe Mossos d���Esquadra, the police force of the Catalan autonomous communityin Spain. The case is described in a comprehensive way (includingpolicies in all relevant policy areas: recruitment, retention, and promotion)and at the same time analyzed with a new analytical framework (includingthe definition of diversity, the motivation for diversity within the organisation,and the facilitation of diversity within the organisation with policies).The goal of the chapter is twofold. First, offer a deeper understanding ofthe dynamics of diversity within this police force. Second, demonstrate theacademic potential of this new analytical framework.

Affirmative Action in Iowa, September 2005

In accordance with 19B.5 of the Code of Iowa, the 2005 Affirmative Action in Iowa report illustrates the progress made during fiscal year 2005 to balance the State's worforce, the challenges that the State must address and the effort that the Department of Administratie Services must lead in order to remove barriers that limit the hiring, retention and advancement of females, minorities and persons with disabilities in the State's workforce. Highlighted in the report are four departments that initiated proactive and innovative measures to address their workplace equal opportunity, affirmative action and diversity programs. Additionally, the Department of Administrative Services-Human Resources Enterprise outlines its plan to build on its past efforts as well as pursue new initiatives to partner with advocacy groups and reach out to the commuity more directely to enhance employment opportunities for females, minorities and persons with disabilities in State of Iowa employment.

Expression and one-step purification of Plasmodium proteins in dictyostelium.

Nearly full-length Circumsporozoite protein (CSP) from Plasmodium falciparum, the C-terminal fragments from both P. falciparm and P. yoelii CSP and a fragment comprising 351 amino acids of P.vivax MSPI were expressed in the slime mold Dictyostelium discoideum. Discoidin-tag expression vectors allowed both high yields of these proteins and their purification by a nearly single-step procedure. We exploited the galactose binding activity of Discoidin Ia to separate the fusion proteins by affinity chromatography on Sepharose-4B columns. Inclusion of a thrombin recognition site allowed cleavage of the Discoidin-tag from the fusion protein. Partial secretion of the protein was obtained via an ER independent pathway, whereas routing the recombinant proteins to the ER resulted in glycosylation and retention. Yields of proteins ranged from 0.08 to 3 mg l(-1) depending on the protein sequence and the purification conditions. The recognition of purified MSPI by sera from P. vivax malaria patients was used to confirm the native conformation of the protein expressed in Dictyostelium. The simple purification procedure described here, based on Sepharose-4B, should facilitate the expression and the large-scale purification of various Plasmodium polypeptides.

Biokinetics and dosimetry of 111 In-DOTA-NOC-ATE compared with 111In-DTPA-Octreotide

Aim: Biokinetics and dosimetry of 111In-DOTA-NOC-ATE (NOCATE) and 111In-DTPA-octreotide (Octreoscan?, OCTREO) were comparatively studied in the same patients. Patients and Methods: Seventeen patients (10 males, 7 females), mean age 60 years referred for an Octreoscan? because of carcinoid (N=9), unspecified neurodendocrine tumors (N=6), thymoma (N=1) or medullary thyroid carcinoma (N=1) accepted a second study with NOCATE. Four patients had no detectable tumor at the time of scanning. Whole-body (WB) anterior-posterior scans were recorded 0.5 (100% reference scan), 4, 24 and 48 hrs (N=17) and 120 hrs (N=6) after injection. OCTREO (178��15 MBq) preceded NOCATE (108��14 MBq) imaging with 16��5 days in 16 patients while 1 patient had first NOCATE followed 14 days later by OCTREO. Blood samples were taken 5, 15, 30, 60, 240 and 1440 min after injection. Background corrected geometric mean counts of WB, lung, kidney, liver, spleen and blood counts expressed in % of the initial composite WB and blood counts, respectively were fitted to bi- or single exponential curves and dosimetry was performed for male and female patients using MIRDOSE3.1 and OLINDA/EXM. Results: Initially, WB, lung and kidney activity was similar but retention was significantly higher for NOCATE compared with OCTREO. Liver and spleen uptake of NOCATE was higher from beginning (p<0.001) and remained so over time. Activity in rest of body showed similar &#945; and &#946; half-lives, but the &#946; half-life fraction of NOCATE was much higher than OCTREO (49% vs. 19%, respectively). Blood T1/2&#946; was longer for NOCATE compared with OCTREO (19 vs. 6h). Residence times were similar in male and female patients while they were in both genders higher for NOCATE than OCTREO. Consequently, effective dose (ED) for NOCATE (ED 114 and 134 &#956;Sv/MBq for man and women, respectively) exceeded that of OCTREO (ED = 61 and 71 &#956;Sv/MBq), the latter results being close to the ICRP-published radiation dose of OCTREO (ED = 54 and 71 ��Sv/MBq, respectively). Differential activity measurement in blood cells and plasma showed that only a minor fraction of NOCATE and OCTREO (<5 % in the mean) was bound to globular blood components. Conclusions: NOCATE showed higher retention in normal organs and delivered roughly twice the radiation dose of OCTREO. The ED of OCTREO in these patients was similar to ICRP80 report when adopting a bladder voiding interval of 2 hours.

Work in inflammatory and degenerative joint diseases

This article focuses on work disability and sick leave and their cost; it also discusses the value of vocational rehabilitation programmes in rheumatic conditions such as rheumatoid arthritis, ankylosing spondylitis, hip and knee osteoarthritis. It acknowledges the importance of work not only for the worker who has one of these diseases but also for the public purse. Much can be done to improve the health of the persons and reduce their disability and its impact in the workplace which will have an important effect on their and their family's quality of life. It is important that neither rehabilitation nor vocational rehabilitation are regarded as bolt-on activities after drug treatment but are seen as an integral part of effective management. Publications dealing with return to work are relatively common in rheumatoid arthritis, less common in ankylosing spondylitis and relatively rare in osteoarthritis. Vocational rehabilitation programmes should aim to facilitate job retention or, failing that, to improve the ability to return to work. The process must be started with in the health arena and it has to be recognised that slow or poor practice in the health service can jeopardise the patient's work potential

Colon-specific deletion of epithelial sodium channel causes sodium loss and aldosterone resistance.

Aldosterone promotes electrogenic sodium reabsorption through the amiloride-sensitive epithelial sodium channel (ENaC). Here, we investigated the importance of ENaC and its positive regulator channel-activating protease 1 (CAP1/Prss8) in colon. Mice lacking the &#945;ENaC subunit in colonic superficial cells (Scnn1a(KO)) were viable, without fetal or perinatal lethality. Control mice fed a regular or low-salt diet had a significantly higher amiloride-sensitive rectal potential difference (&#8710;PDamil) than control mice fed a high-salt diet. In Scnn1a(KO) mice, however, this salt restriction-induced increase in &#8710;PDamil did not occur, and the circadian rhythm of &#8710;PDamil was blunted. Plasma and urinary sodium and potassium did not change with regular or high-salt diets or potassium loading in control or Scnn1a(KO) mice. However, Scnn1a(KO) mice fed a low-salt diet lost significant amounts of sodium in their feces and exhibited high plasma aldosterone and increased urinary sodium retention. Mice lacking the CAP1/Prss8 in colonic superficial cells (Prss8(KO)) were viable, without fetal or perinatal lethality. Compared with controls, Prss8(KO) mice fed regular or low-salt diets exhibited significantly reduced &#8710;PDamil in the afternoon, but the circadian rhythm was maintained. Prss8(KO) mice fed a low-salt diet also exhibited sodium loss through feces and higher plasma aldosterone levels. Thus, we identified CAP1/Prss8 as an in vivo regulator of ENaC in colon. We conclude that, under salt restriction, activation of the renin-angiotensin-aldosterone system in the kidney compensated for the absence of ENaC in colonic surface epithelium, leading to colon-specific pseudohypoaldosteronism type 1 with mineralocorticoid resistance without evidence of impaired potassium balance.

Downsizing, Job Insecurity, and Firm Reputation

This paper studies how firms make layoff decisions in the presence of adverse shocks. In this uncertain environment, workers' expectations about their job security affect their on-the-job performance. This productivity effect on job insecurity forces firms to strike a balance between laying off redundant workers and maintaining survivors' commitment when deciding on the amount and timing of downsizing. This framework offers an explanation of conservative employment practices (such as zero or reduced layoffs) based on firms having private information about their future profits. High retention rates and wages can signal that the firm has a bright future, boosting workers' confidence. Moreover, the model provides clear predictions about when waves of downsizing will occur as opposed to one-time massive cuts.

Iowa's Adult Basic Education Program Annual Benchmark Report Executive Summary, 2002

The purpose of this publication is to present the Executive Summary for the Program Year 2002 report on Iowa���s adult basic education program benchmarks. The passage of the Workforce Investment Act (WIA) of 1998 [Public Law 105-220] by the 105th Congress has ushered in a new era of collaboration, coordination, cooperation and accountability. The overall goal of the Act is ���to increase the employment, retention, and earnings of participants, and increase occupational skill attainment by participants, and, as a result improve the quality of the workforce, reduce welfare dependency, and enhance the productivity and competitiveness of the Nation.���

Iowa's Adult Basic Education Program Annual Benchmark Report Executive Summary, 2001

The purpose of this publication is to present the Executive Summary of the first annual report on the results of Iowa���s adult basic education program benchmarks. The passage of the Workforce Investment Act (WIA) of 1998 [Public Law 105-220] by the 105th Congress has ushered in a new era of collaboration, coordination, cooperation and accountability. The overall goal of the Act is ���to increase the employment, retention, and earnings of participants, and increase occupational skill attainment by participants, and, as a result improve the quality of the workforce, reduce welfare dependency, and enhance the productivity and competitiveness of the Nation.���

Iowa Adult Literacy Program Annual Benchmark Report, 2005

The purpose of this publication is to present the Program Year 2005 report on Iowa���s adult literacy program benchmarks. The passage of the Workforce Investment Act (WIA) of 1998 [Public Law 105-220] by the 105th Congress ushered in a new era of collaboration, coordination, cooperation and accountability. The overall goal of the Act is ���to increase the employment, retention, and earnings of participants, and increase occupational skill attainment by participants, and, as a result improve the quality of the workforce, reduce welfare dependency, and enhance the productivity and competitiveness of the Nation.���

Iowa's Adult Basic Education Program Annual Benchmark Report Executive Summary, 2005

The purpose of this publication is to present the Executive Summary for the Program Year 2002 report on Iowa���s adult basic education program benchmarks. The passage of the Workforce Investment Act (WIA) of 1998 [Public Law 105-220] by the 105th Congress has ushered in a new era of collaboration, coordination, cooperation and accountability. The overall goal of the Act is ���to increase the employment, retention, and earnings of participants, and increase occupational skill attainment by participants, and, as a result improve the quality of the workforce, reduce welfare dependency, and enhance the productivity and competitiveness of the Nation.���