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The Millennial generation is expected to comprise nearly 34% of the labor force in 2024 (U.S. Census Bureau, 2012). A significant catalyst for the growth pattern is exhibited by Baby Boomer cohort retirement and deaths among them in the United States (Pew Research Center, 2016). The majority of the Baby Boomer generation is historically connected with public sector work, but the composition of the generational workplace/workforce has been altered to exhibit diverse needs and contributions. The Millennial cohort has characteristics that directly align with the premise of Public Sector Organizations. However, other attributes fail to align with the Public Sector Organization's core characteristic of bureaucracy. Based on the Bureau of Labor Statistics (2014) the Millennial generation only made up 24.5% of government or public sector employees, compared to 33.7% in the private sector, and this statistic continues to be contemporarily disproportionate. Therefore, the purpose of this study is to identify and assess Public Sector Management strategies and interventions that would increase retention among the Millennial cohort in Public Sector Organizations. A qualitative study using a Thematic Analysis was done to examine whether a productive relationship between the United States Public Sector Organization and Millennial generation characteristics could increase retention. Based on an analysis of the research, the study finds that there is a significant relationship between the Millennial generation and the Public Sector Organization, which may cause an increase in retention if Public Sector Management aligns certain Public Sector Organization attributes to meet the needs of the Millennial generation.

Infection with human rhinovirus (HRV) is thought to result in acute respiratory exacerbations of chronic obstructive pulmonary disorder (COPD). Consequently, prevention of HRV infection may provide therapeutic benefit to these patients. As all major group HRV serotypes infect cells via an interaction between viral coat proteins and intercellular adhesion molecule-1 (ICAM-1), it is likely that inhibitors of this interaction would prevent or reduce infections. Our objective was to use phage display technology in conjunction with naive human antibody libraries to identify anti-ICAM-1 antibodies capable of functional blockade of HRV infection. Key to success was the development of a robust, functionally relevant high-throughput screen (HTS) compatible with the specific challenges of antibody screening. In this article, we describe the development of a novel homogeneous time-resolved fluorescence (HTRF) assay based on the inhibition of soluble ICAM-1 binding to live HRV16. We describe the implementation of the method in an antibody screening campaign and demonstrate the biological relevance of the assay by confirming the activity of resultant antibodies in a cell-based in vitro HRV infection assay.

Reactions of isolated ion pairs in solution have been modelled using the Debye–Smoluchowski equation for diffusion and conduction. An activation step was incorporated using a partially reflecting boundary condition. The method of matched expansions and the Abelian theorem of Laplace transforms was used to give an approximate solution of the Debye–Smoluchowski equation. Numerical integrations based on the finite‐difference method confirmed these approximate analytic formulas.

Sustainable energy supplies are needed to supplement and eventually replace fossil fuels. Molecular hydrogen H2 is a clean burning, high-energy fuel that is also used as reducing gas in industrial processes. H2 is mainly synthesized by steam reforming of natural gas, a non-renewable fuel. There are biosynthetic strategies for H2 production; however, they are associated with poor yield and have high cost. The application of an electrochemical driving force in a microbial electrolysis cell (MEC) improves the yield of biological reactions. The performance of the MEC is influenced by experimental parameters such as the electrode material, reactor design, microbial consortia and the substrate. In this review, factors that affect the performance of MECs are discussed and critically analysed. The potential for scale-up of H2 bioelectrosynthesis is also discussed.

In response to infections and irritants, the respiratory epithelium releases the alarmin interleukin (IL)-33 to elicit a rapid immune response. However, little is known about the regulation of IL-33 following its release. Here we report that the biological activity of IL-33 at its receptor ST2 is rapidly terminated in the extracellular environment by the formation of two disulphide bridges, resulting in an extensive conformational change that disrupts the ST2 binding site. Both reduced (active) and disulphide bonded (inactive) forms of IL-33 can be detected in lung lavage samples from mice challenged with Alternaria extract and in sputum from patients with moderate–severe asthma. We propose that this mechanism for the rapid inactivation of secreted IL-33 constitutes a ‘molecular clock' that limits the range and duration of ST2-dependent immunological responses to airway stimuli. Other IL-1 family members are also susceptible to cysteine oxidation changes that could regulate their activity and systemic exposure through a similar mechanism., IL-33, released by epithelial cells in response to stress, is a potent activator of inflammation. Here Cohen et al. show that secreted IL-33 is rapidly inactivated by disulfide bond formation that prevents binding to its receptor, and that IL-33-related cytokines are susceptible to similar oxidation.

Background: In response to infections and irritants, the bronchial epithelium releases the alarmin interleukin(IL)-33 to elicit a rapid immune response. Regulation of IL-33 following release from the cell is poorly understood. Here we characterise the lifecycle and isoforms of the IL-33 protein in vivo. Methods: 25ug of Alternaria extract was administered intranasally to BALB/c mice to release endogenous IL-33 into BAL Fluid. Human endogenous IL-33 was derived from lung explants or sputum from asthma patients. Recombinant IL-33 WT or mutant proteins were made in E. Coli . IL-33 proteins were characterised by Western Blot, mass spectrometry, ELISA, ST2-dependent functional assays and in vivo responses. Results: Mature IL-33 was rapidly oxidised in cell culture media, serum and the in vivo lung to generate an isoform with two disulphide bridges, C208–C259 and C227–C232, respectively. This resulted in an extensive conformational change with disruption of the ST2 binding site that terminated activity. A cysteine to serine mutant form of IL-33 was resistant to oxidation and was 30-fold more potent both in vitro and in vivo . Disulphide-bonded inactive IL-33 was detected following the peak of active IL-33 in lung lavage samples from animals challenged with Alternaria extract and from human lung explants. Disulphide bonded IL-33 was the most commonly detected isoform in sputum from patients with moderate-severe asthma. Conclusion: We propose oxidation as a novel mechanism for the rapid inactivation of secreted IL-33 that limits the range and duration of ST2-dependent immunological responses to airway stimuli.

Laboratory investigations play an important role in the diagnosis of rheumatic disorders but many tests are of limited specificity. It is therefore important to select tests in the light of careful clinical assessment rather than blindly sending off a battery of requests, as the results may be confusing. We review the frequency of autoantibodies in different disorders and their value for diagnosis and, in some cases, for prognosis and monitoring. We also review the relative merits and disadvantages of plain radiographs, ultrasound, isotope, CT and MR imaging for different types of musculoskeletal problem. Finally we describe the typical imaging characteristics of degenerative and inflammatory disorders including rheumatoid arthritis and seronegative spondyloarthropathies in the peripheral joints and spine.

A combined clinical and radiological approach is essential in the diagnosis and management of rheumatological disorders and their complications. There is significant overlap between the radiological features of several rheumatic disorders; therefore, close collaboration is required between clinician and radiologist when selecting the appropriate imaging modality and interpreting the result.

Biologics represent a fast-growing class of therapeutics in the pharmaceutical sector. Discovery of therapeutic antibodies and characterization of peptides can necessitate high expression of the target gene requiring the generation of clonal stably transfected cell lines. Traditional challenges of stable cell line transfection include gene silencing and cell-to-cell variability. Our inability to control these can present challenges in lead isolation. Recent progress in site-specific targeting of transgene to specific genomic loci has transformed the ability to generate stably transfected mammalian cell lines. In this article, we describe how the use of the Jump-In platform (Life Technologies, Carlsbad, CA) has been applied to drug discovery projects. It can easily and rapidly generate homogeneous high-expressing cell pools with a high degree of reproducibility. Their use in cell-based screening to identify specific binders, identify binding to relevant species variants, or detect functionally relevant therapeutic antibodies is central in driving drug discovery.

Previous studies have shown sequence similarity between a region of the autosomal dominant polycystic kidney disease (ADPKD) protein, polycystin-1 and a sea urchin sperm glycoprotein involved in fertilization, the receptor for egg jelly (suREJ). We have analysed sequence databases for novel genes encoding PKD/REJ-like proteins and found a significant region of homology to a large open reading frame in genomic sequence from human chromosome 22. Northern analysis showed that this is a functional gene [termed the polycystic kidney disease and receptor for egg jelly related gene ( PKDREJ )], but unlike polycystin-1, has a very restricted expression pattern; the approximately 8 kb transcript was found exclusively in testis, coincident with the timing of sperm maturation. The PKDREJ transcript was cloned by screening a testis cDNA library and RT-PCR which revealed a 7660 bp mRNA terminating with a 900 bp 3'UTR and a polyA tail. Comparison with genomic sequence showed that PKDREJ is intronless; sequencing the mouse orthologue revealed a similar structure. The predicted human PKDREJ protein has 2253 amino acids (calculated molecular mass 255 kDa) and sequence similarity over approximately 2000 amino acids with polycystin-1, corresponding to the predicted membrane associated region and the area of homology ( approximately 1000 amino acids) with the suREJ protein (the REJ module). The suREJ protein binds the glycoprotein coat of the egg (egg jelly), triggering the acrosome reaction, which transforms the sperm into a fusogenic cell. The sequence similarity and expression pattern suggests that PKDREJ is a mammalian equivalent of the suREJ protein and therefore may have a central role in human fertilization.

The critical role played by IgE in allergic asthma is well-documented and clinically precedented, but some patients in whom IgE neutralization may still offer clinical benefit are excluded from treatment with the existing anti-IgE therapy, omalizumab, due to high total IgE levels or body mass. In this study, we sought to generate a novel high affinity anti-IgE antibody (MEDI4212) with potential to treat a broad severe asthma patient population. Analysis of body mass, total and allergen-specific IgE levels in a cohort of severe asthmatics was used to support the rationale for development of a high affinity IgE-targeted antibody therapeutic. Phage display technology was used to generate a human IgG1 lead antibody, MEDI4212, which was characterized in vitro using binding, signaling and functional assay systems. Protein crystallography was used to determine the details of the interaction between MEDI4212 and IgE. MEDI4212 bound human IgE with an affinity of 1.95 pM and was shown to target critical residues in the IgE Ce3 domain critical for interaction with FceRI. MEDI4212 potently inhibited responses through FceRI and also prevented the binding of IgE to CD23. When used ex vivo at identical concentration, MEDI4212 depleted free-IgE from human sera to levels ~1 log lower than omalizumab. Our results thus indicate that MEDI4212 is a novel, high affinity antibody that binds specifically to IgE and prevents IgE binding to its receptors. MEDI4212 effectively depleted free-IgE from human sera ex vivo to a level (1 IU/mL) anticipated to provide optimal IgE suppression in severe asthma patients.

The discoidin domain receptor (DDR) is a new class of receptor tyrosine kinase that is distinguished by a unique extracellular domain homologous to the lectin Discoidin I found Dictyostelium discoideum. A cosmid was isolated from a human chromosome 6 cosmid library containing the DDR gene. A complete genomic contig of the DDR gene was constructed from seven subclones of the cosmid. The cosmid fragments were analyzed by PCR, sequencing, and comparison of genomic/cDNA sequence. The DDR gene is composed of 17 exons, ranging in size from 96 to 1014 bp, distributed along approximately 12 kb of genomic DNA. The extracellular domain is encoded by 8 exons of which three code for the discoidin domain. The transmembrane domain is encoded by 1 exon, the juxtamembrane by 3 exons, and the catalytic domain by 5 exons. The generation of the two splice variants of DDR, EDDR1 and EDDR2 are explained by the genomic structure. Exon 11 (111 bp in the juxtamembrane domain) is present in DDR and absent in the splice variant EDDR1. An inverted repeat of 20 bp was identified at the 3' exon-intron junction of exon 11, which results in a lariat loop-like secondary structure. EDDR2 is generated because of a cryptic splice acceptor site that results in an extra 18 bp (6 amino acids) inserted 5' of exon 14 in the catalytic domain. A polymorphic (GT)17 repeat was identified in intron 5 with a heterozygosity of 0.71. The exon-intron structure of the DDR gene will be helpful in further understanding of its function and explains the possible structural basis for the two splice variants.

Comprises a speech given by Sir Robin Butler, Secretary to the Cabinet and head of the Home Civil Service, at a conference held in London in November 1995 to celebrate the twenty‐fifth anniversary of the Civil Service College. Draws from the speaker’s own experiences of working in the Civil Service and details the changes leading to the opening of the college in 1970, which provides high quality training and consultancy in management and policy issues, and has gained international recognition. Considers the main themes of the reforms in the management of the Civil Service, not just in the UK, but also in the USA and New Zealand, and highlights the Civil Service’s emphasis on well‐trained, knowledgeable and professional staff.

(1995). The Themes of Public Service Reform in Britain and Overseas. Policy Studies: Vol. 16, No. 3, pp. 4-25.

While the publication of ‘cult’ texts has become somewhat common place in the management world it is much more of a rarity in public administration. However, the success in the United States of David Osborne and Ted Gaebler's Reinventing Government 1992) may be a pointer to the emergence of an identifiable and coherent field of public management offering analysis and prescriptions in tune with management reform in government on both sides of the Atlantic. In the symposium that follows we offer three different perspectives on ‘Reinventing Government’. In the first, Sir Robin Butler indicates how the major themes of the book can be seen to correspond with many of the recent management initiatives in UK government. In the second, Grant Jordan raises some questions on the assumptions made in Osborne and Gaebler's thesis and questions some of their prescriptions. In the final article, Rod Rhodes describes some alternative academic perspectives which he believes could inform the reform of government debate and asks what makes a cult best seller.

This is a slightly edited version of a lecture Sir Robin Butler gave in the FDA series on "The Future of the Civil Service" on October 15th, 1991. Public Policy and Administration is grateful to Sir Robin for allowing us to publish the full version of his lecture. An edited version appeared in FDA News, November 1991.

Bone mineral content (BMC) of the distal forearm was measured by single photon absorptiometry in 142 patients with rheumatoid arthritis (RA) of whom 27/54 men and 44/88 women received low-dose steroid therapy (less than 10 mg/day). To study the effect of steroid therapy a case-control analysis was undertaken in patients matched for age, sex and disease duration. Steroid therapy was associated with a reduced BMC in men (1.16 +/- 0.29 versus 1.32 +/- 0.23; P less than 0.05) and post-menopausal (0.76 +/- 0.24 versus 0.91 +/- 0.25; P less than 0.02) but not pre-menopausal women (1.1 +/- 0.28 versus 1.1 +/- 0.17). Symptomatic fractures were more common in steroid-treated patients than in those who had not received steroids (10/71 versus 2/71; P less than 0.05). Serum osteocalcin, an index of bone formation, was measured in 106 cases. It tended to be higher in patients with RA than in controls but the values observed in steroid and non-steroid RA groups did not differ significantly. We conclude that low-dose steroid therapy is associated with increased bone loss and numbers of fractures in patients with RA but this does not appear to be the result of a simple defect in bone formation.

X-linked dystonia-parkinsonism (XDP) is a recessive disorder characterized by generalized dystonia with some patients exhibiting parkinsonism. The disease gene, DYT3, is located between DXS453 (DXS993) and DXS559, and strongest linkage disequilibrium is found distal to DXS7117 and proximal to DXS559. We have isolated and analyzed four novel polymorphic markers between DXS7117 and DXS559 and, by haplotype analysis, have narrowed the candidate interval to

A second gene for autosomal dominant polycystic kidney disease (ADPKD), PKD2, has been recently identified. Using antisera raised to the human PKD2 protein, polycystin-2, we describe for the first time its distribution in human fetal tissues, as well as its expression in adult kidney and polycystic PKD2 tissues. Its expression pattern is correlated with that of the PKD1 protein, polycystin-1. In normal kidney, expression of polycystin-2 strikingly parallels that of polycystin-1, with prominent expression by maturing proximal and distal tubules during development, but with a more pronounced distal pattern in adult life. In nonrenal tissues expression of both polycystin molecules is identical and especially notable in the developing epithelial structures of the pancreas, liver, lung, bowel, brain, reproductive organs, placenta, and thymus. Of interest, nonepithelial cell types such as vascular smooth muscle, skeletal muscle, myocardial cells, and neurons also express both proteins. In PKD2 cystic kidney and liver, we find polycystin-2 expression in the majority of cysts, although a significant minority are negative, a pattern mirrored by the PKD1 protein. The continued expression of polycystin-2 in PKD2 cysts is similar to that seen by polycystin-1 in PKD1 cysts, but contrasts with the reported absence of polycystin-2 expression in the renal cysts of Pkd2+/- mice. These results suggest that if a two-hit mechanism is required for cyst formation in PKD2 there is a high rate of somatic missense mutation. The coordinate presence or loss of both polycystin molecules in the same cysts supports previous experimental evidence that heterotypic interactions may stabilize these proteins.

The RNA polymerase sigma factor sigma N (sigma 54) is characterized by the presence, near the C-terminal end of the protein, of a highly conserved sequence of 10 amino acids (ARRTVAKYRE) that has been termed the RpoN box. In order to examine the function of this motif, which is predicted to adopt an alpha-helical structure, we have isolated a number of mutations that alter residues within the box and examined the properties of the sigma N derivatives encoded by them. Certain mutations that alter charged and potentially exposed residues within the motif result in transcriptionally inactive proteins with impaired promoter recognition but no impairment in core RNA polymerase binding. We therefore suggest that the RpoN box could play a direct or indirect role in recognition of the -24, -12 promoter consensus that is characteristic of sigma N-dependent genes.

A protein receptor tyrosine kinase (EDDR1) has been isolated from a complementary DNA library of SKOV-3, an epithelial ovarian cancer cell line. The primary structure of the predicted amino acid sequence of the protein shows a novel N-terminal region that has homology to a factor VIII-like domain. The C-terminal catalytic domain has all of the canonical sequence motifs of a receptor tyrosine kinase with homology to the TRK-2H protein (49%), which suggests that it is a type II receptor. It is expressed in epithelial cells of several tissues. To determine the chromosomal localization of the gene, somatic cell hybrids were analyzed by PCR amplification using oligonucleotide primers specific for EDDR1. Segregation was observed to a hybrid containing human chromosome 6. Cosmids for EDDR1 were isolated from a human chromosome 6 cosmid library and were shown by fluorescence in situ hybridization to map to 6q16.

Fick's second diffusion equation, with an added exponential sink term, is integrated numerically to simulate the decay of electrons at short times in the presence of scavengers. The time dependence of the scavenger concentration profile, the scavenging rate constant, and the electron concentration are illustrated graphically. Using the experimental results of Buxton et al. and Jonah et al. It is shown that the Smoluchowski equation is valid within their experimental time ranges provided the cage encounter distance is replaced by Reff, where Reff can be evaluated explicitly in terms of reaction parameters. It is also shown that tunnelling from relaxed traps may make a significant contribution to ultra-short time electron scavenging.

The diffusion equation, based on Fick's second law, has been solved numerically with an added sink term, which falls off as r−6, to simulate reaction by a dipole—dipole mechanism. The results show that, even for large Ro values (≈ 6 nm), the Forster equation is seriously in error for diffusion coefficients > 10−9 m2 s−1, but that an interpolation formula, proposed by Gosele et al., reproduces the numerical solutions with high precision.

The quenching of triplet phenanthiene by Cu 2+ has been studied over a range of high viscosities. The rate constants were determined in the time-dependent regime and are compatible with an encounter distance which increases with increasing viscosity. The Smoluchowski analysis of the time-dependence breaks down at very high viscosities in agreement with a numerical analysis.

Imaging in rheumatology was in the past largely confined to radiographs of the hands and sacroiliac joints (SIJs) helping to establish the diagnosis and then monitoring disease progression. Radiographs are not very sensitive for early inflammation in inflammatory rheumatic disorders and the demand on imaging services was therefore limited. However, over the last 10–15 years new drugs and new technologies have brought new challenges and opportunities to rheumatology and radiology as specialties. New drug treatments allow more effective treatment, preventing many complications. Early diagnosis and disease monitoring has become the challenge for the rheumatologist and radiologist alike. The best possible patient outcome is only achieved if the two specialties understand each other’s viewpoint. This article reviews the role of imaging—in particular radiography, magnet resonance imaging, computer tomography, ultrasound and nuclear medicine—for the diagnosis and monitoring of rheumatological disorders, concentrating on rheumatoid arthritis, inflammatory spondylarthropathies and gout. Teaching Points • New drugs for the treatment of inflammatory disorders has led to greatly improved outcomes. • Imaging often allows for earlier diagnosis of inflammatory disorders. • Early diagnosis and treatment can often prevent the development of crippling disease manifestations. • Tailored imaging examinations are best achieved by consultation of rheumatologist and radiologist.

Protein tyrosine kinases play an imporH-RYK, an Unusual Receptor Kinase: Isolation and Analysis of Expression in Ovarian Cancertant role in cellular metabolism as key components of signal transduction pathways. They are involved in cellular growth, differentiation, and development. Receptor tyrosine kinases (EGF receptor and c-erbB2) have been shown to be important in the pathogenesis of cancer. In ovarian cancer, overexpression of c-erbB2, a type I receptor, has been correlated with an adverse effect on survival of patients. An unusual receptor tyrosine kinase, H-RYK, has been isolated from a complimentary DNA library of SKOV-3, an epithelial ovarian cancer cell line, using a polymerase chain reaction-mediated approach. The primary structure of the predicted amino acid sequence of the protein shows a novel NH2-terminal region. The catalytic region shows homology to other tyrosine kinases, the closest homology being with v-sea(39%). A significant alteration in the catalytic domain is that the highly conserved “DFG” triplet in subdomain VII is altered to “DNA.” The gene was mapped to chromosome 3q22. A single transcript of 3.0 kb is expressed in heart, brain, lung, placenta, liver, muscle, kidney, and pancreas by Northern analysis with maximal expression in skeletal muscle. In situ hybridization analysis on human tissues demonstrated localization of message in the epithelial and stromal compartment of tissues such as brain, lung, colon, kidney, and breast. There was minimal to absent expression of H-RYK on surface epithelium of ovaries. In benign (3) and borderline tumors of the ovary (5), there was expression in the stromal compartment. However, in malignant tumors (24) there was increased expression predominantly confined to the epithelium. Polyclonal antisera raised against synthetic peptides recognize a 100-kD protein in ovarian cancer cells and other cell lines. In contrast to other receptor tyrosine kinases, the receptor did not phosphorylate in an in vitro kinase assay. The expression of this unusual receptor tyrosine kinase in epithelial ovarian cancer suggests that it may be involved in tumor progression, which needs further investigation.

The diffusion equation, derived from Fick's second law, with an added exponential sink term to simulate electron tunnelling, is integrated numerically to determine the rate of electron decay at times greater than 1 ps. The effect of a coulomb interaction with a charged scavenger is examined and the steady-state rate constant shown to approximate closely to that obtained by combining the separate effects of tunnelling and charge-affected diffusion, which can be expressed analytically. Diffusion in the presence of a charge-induced dipole interaction is investigated for the case of scavenging of localised electrons in alkanes. The rate constant is shown to be dominated by random diffusion and tunnelling and the bias induced by the interaction is of little consequence. The sensivity of the rate constant to changes in the pre-exponential factor in the sink term is shown to be most favourable at short times.