Traditional Restriction Research Articles

Therapeutic Options in Managing Hyponatremia: Focus on Arginine Vasopressin Receptor Antagonists

Purpose To describe the available and emerging treatment options for acute and chronic hyponatremia, including the efficacy, safety, and recommendations regarding appropriate use, monitoring, and treatment individualization. Summary Arginine vasopressin (AVP) receptor antagonists provide an opportunity to address some of the unmet medical needs of patients with hyponatremia. Traditional therapies, including diuretics, fluid restriction, and saline infusions, have variable effects, potential toxicities, and issues with patient adherence. Furthermore, these therapies are not specific to the underlying pathophysiology causing the hyponatremia. The recently approved AVP receptor antagonists target the underlying abnormal release of AVP that is very likely at the core of the physiology. Conclusion Management of hyponatremia requires balancing the benefits of therapeutic intervention for the restoration of normal serum sodium against the potential risks. The data available indicate that this new class of medications, the AVP receptor antagonists, can favorably affect serum sodium and clinical outcomes in patients with hypervolemic and euvolemic hyponatremia.

A cleaved amplified polymorphic sequence (CAPS) method for the identification of geographical isolates of Schistosoma japonicum in China.

Schistosomiasis japonica, caused by the blood fluke Schistosoma japonicum, is a severe parasitic zoonosis, causing significant morbidity, mortality and considerable economic losses in south-east Asia, particularly in mainland China (Zhou et al., 2005; Bergquist et al., 2008; Zhou et al., 2008; Li et al., 2010). Although there have been significant efforts to control schistosomiasis in China, this disease appears to be re-emerging due to climate changes and human activities (Wang et al., 2009). Thus far, schistosomiasis is still endemic in seven provinces (Yunnan, Sichuan, Anhui, Hubei, Jiangxi, Jiangsu and Hunan provinces) of China (Ross et al., 2001), and the impact of this communicable disease in China is almost equal to that of HIV/AIDS and tuberculosis (Engels et al., 2005). In previous studies, genetic variation among S. japonicum populations from different endemic provinces in China has been detected using a variety of genetic markers (Bogh et al., 1999; Sorensen et al., 1999; Zhu et al., 1999; Shrivastava et al., 2005; Zhao et al., 2009a; Zhao et al., 2009b; Zhao et al., 2009c). However, these markers and the detected genetic variability have not been utilized for the practical identification and differentiation of S. japonicum geographical isolates in China. The economic reform and the deterioration of ecological environment, flash floods, landslides and the mobility of human population appear to be having an impact on the transmission of S. japonicum (Li et al., 2000; Seto et al., 2008). Consequently, it is important to be able to identify different genetic variants of S. japonicum in China. Various methods for the analysis of genetic variation have been developed, including single-strand conformation polymorphism (Simoes et al., 2007), denaturing high-performance liquid chromatography (Nickerson et al., 2000), gene chips (Schmalzing et al., 2000), TaqMan probe (Shi et al., 1999) and pyrosequencing technology (Ahmadian et al., 2000). However, such techniques can be time-consuming and/or require detection instruments or labelled oligonucleotides. In contrast, the cleaved amplified polymorphism sequence (CAPS) technique is a simple and effective method to allow the rapid detection of single nucleotide polymorphisms (SNPs) (Neff et al., 1998; Komori and Nitta, 2005). CAPS is based on the PCR-based amplification of targeted genes and subsequent digestion of amplicons using restriction enzymes. The digested PCR products are separated by agarose gel electrophoresis. CAPS generates the same type of data as traditional restriction fragment length polymorphism analysis, but significantly reduces the amount of purified DNA required for analysis and time-consuming steps. This method can detect both homozygous and heterozygous individuals (Weiland and Yu, 2003; Komori and Nitta, 2005). CAPS has been successfully used to study genetic diversity in plants, such as Arabidopsis thaliana (Hardtke et al., 1996; Barth et al., 2002), Cryptomeria japonica (Tsumura and Tomaru, 1999; Tsumura et al., 1999) and Pisum sativum L. (Konovalov et al., 2009), but has seldom been used to study variation in parasites (Gandhi et al., 2009). The aims of the present study were to identify SNP sites specific for geographical isolates of S. japonicum based on results of previous studies. Using these SNPs, the CAPS technique was developed and used to identify and differentiate S. japonicum isolates from Yunnan province and those from other endemic provinces.

A Gateway MultiSite Recombination Cloning Toolkit

The generation of DNA constructs is often a rate-limiting step in conducting biological experiments. Recombination cloning of single DNA fragments using the Gateway system provided an advance over traditional restriction enzyme cloning due to increases in efficiency and reliability. Here we introduce a series of entry clones and a destination vector for use in two, three, and four fragment Gateway MultiSite recombination cloning whose advantages include increased flexibility and versatility. In contrast to Gateway single-fragment cloning approaches where variations are typically incorporated into model system-specific destination vectors, our Gateway MultiSite cloning strategy incorporates variations in easily generated entry clones that are model system-independent. In particular, we present entry clones containing insertions of GAL4, QF, UAS, QUAS, eGFP, and mCherry, among others, and demonstrate their in vivo functionality in Drosophila by using them to generate expression clones including GAL4 and QF drivers for various trp ion channel family members, UAS and QUAS excitatory and inhibitory light-gated ion channels, and QUAS red and green fluorescent synaptic vesicle markers. We thus establish a starter toolkit of modular Gateway MultiSite entry clones potentially adaptable to any model system. An inventory of entry clones and destination vectors for Gateway MultiSite cloning has also been established (www.gatewaymultisite.org).

Current Therapeutic Options for Hyponatremia: Indications, Limitations, and Confounding Variables

The arginine vasopressin receptor antagonists, also known as the vaptans, are a new class of agents that address some of the unmet medical needs of patients with hyponatremia. Traditional therapies, including diuretics, fluid restriction, and saline infusions, have variable effects, potential toxicities, and concerns with patient adherence. Furthermore, these therapies are not specific to the underlying pathophysiology causing the hyponatremia. The recently approved arginine vasopressin receptor antagonists, however, target the underlying abnormal release of arginine vasopressin that is very likely at the core of the pathophysiology. Management of hyponatremia requires balancing the benefits of therapeutic intervention to restore normal serum sodium concentrations against the potential risks. Additional clinical experience is needed to develop reliable criteria for determining which patients should be treated with these agents. However, the data available indicate that this new class of drugs can favorably affect serum sodium concentration and clinical outcomes in patients with hypervolemic and euvolemic hyponatremia.

Nanofiltration (NF) membranes: the next generation separators for all vanadium redox flow batteries (VRBs)?

NF membranes, as an alternative to traditional ion exchange membranes, were first proposed and successfully prepared for VRBs based on a totally new concept of tuning the vanadium/proton selectivity viapore size exclusion. The results showed that membranes show increasing vanadium ion/proton (V/H) selectivity with decreasing pore size distribution. VRBs assembled with prepared NF membranes exhibited comparable performance to that of commercialized Nafion. The concept could potentially overcome the traditional restriction from ion exchange membranes and provide much more material options for VRB membranes.

Large neutral amino acids in the treatment of PKU: from theory to practice

Notwithstanding the success of the traditional dietary phenylalanine restriction treatment in phenylketonuria (PKU), the use of large neutral amino acid (LNAA) supplementation rather than phenylalanine restriction has been suggested. This treatment modality deserves attention as it might improve cognitive outcome and quality of life in patients with PKU. Following various theories about the pathogenesis of cognitive dysfunction in PKU, LNAA supplementation may have multiple treatment targets: a specific reduction in brain phenylalanine concentrations, a reduction in blood (and consequently brain) phenylalanine concentrations, an increase in brain neurotransmitter concentrations, and an increase in brain essential amino acid concentrations. These treatment targets imply different treatment regimes. This review summarizes the treatment targets and the treatment regimens of LNAA supplementation and discusses the differences in LNAA intake between the classical dietary phenylalanine-restricted diet and several LNAA treatment forms.

Genotyping the BDNF rs6265 (val66met) polymorphism by one-step amplified refractory mutation system PCR

The brain derived neutrophic factor (BDNF), a 27 kD polypeptide, is one of the most widely expressed neurotrophins in the brain, regulating neural development and plasticity. The BDNF gene contains a functional single-nucleotide polymorphism (rs6265), which results in a valine to methionine substitution (val66met), leading to reduced mature BDNF expression. This polymorphism has been widely implicated in a host of psychiatric disorders and is a focus of many ongoing psychiatric genetic studies. To develop an efficient and rapid method to detect the val66met polymorphism in a one-step PCR reaction. We have designed four PCR primers that amplify the BDNF gene region containing rs6265. The specificity of the four primers in a single PCR reaction amplifies two allele-specific amplicons (253 and 201 bp) and the entire region (401 bp) as an internal control, which are easily distinguished on a polyacrylamide gel. The effectiveness and efficiency of the results are validated by traditional NlaIII restriction enzyme digestion, sequencing of resulting bands and confirmation on 308 genomic DNA samples. This new method describes a rapid, sensitive, cost effective and high throughput genotyping of the BDNF val66met polymorphism, ideal for large-scale genotyping studies.

Alternate-day fasting protects the rat heart against age-induced inflammation and fibrosis by inhibiting oxidative damage and NF-kB activation

The free radical theory of aging is currently one of the most popular. In parallel, many studies have demonstrated the association of fibrosis and increased oxidative stress in the pathogenesis of some chronic human diseases, and fibrosis is often characteristic of aging tissues. One of the few interventions that effectively slow aging is calorie restriction and the protection against the age-associated increase of oxidative stress remains one of the foremost hypotheses to explain this action. As an alternative to traditional calorie restriction, another dietary regimen, termed alternate-day fasting, has also been tested, whose antiaging mechanisms have not been explored so much extensively. We thus studied the effects of alternate-day fasting, started at 2 months of age, on oxidative stress and fibrosis in the heart during aging. In the left ventricle of the heart of elderly (aged 24 months) versus young (aged 6 months) male rats we found a significant increase in oxidative stress paralleled by increased fibrosis. In parallel there was a significant increase in inflammatory cytokine levels and in NF-kB DNA binding activity with advancing age. Alternate-day fasting protected against all these age-related phenomena. These data support the hypothesis that this kind of dietary restriction protects against age-related fibrosis, at least in part by reducing inflammation and oxidative damage, and this protection can thus be considered a factor in the prevention of age-related diseases with sclerotic evolution.

Simple, Fast, and Efficient Cloning of PCR Products with TOPO ® Cloning Vectors

BioTechniquesVol. 46, No. 6 Application Forum - Sponsored PaperOpen AccessSimple, Fast, and Efficient Cloning of PCR Products with TOPO® Cloning VectorsBalwant PatelBalwant PatelLife Technologies, 5791 Van Allen Way, Carlsbad, CA, 92008, USASearch for more papers by this authorPublished Online:25 Apr 2018https://doi.org/10.2144/000113158AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinkedInRedditEmail IntroductionTraditional restriction enzyme cloning methods are time-consuming and tedious, and with their relatively low efficiencies, multiple colonies must be screened to find the insert of interest. By removing laborious and unreliable steps from standard cloning procedures, TOPO® PCR cloning enables benchtop cloning reactions in 5 min and obtains up to 95% recombinants (Table 1). The use of TOPO® cloning vectors has been reported in the literature more than 10,000 times since the technology was launched.Table 1– Efficient subcloning with TOPO® cloning kits.Various PCR products were cloned into the pCR®2.1-TOPO® vector, transformed into One Shot®TOP10 Chemically Competent E. coli, and plated onto X-gal/amp plates. The percent recombinants represent the numbers of white colonies/the number of total colonies.95% Cloning Efficiency in Three StepsInvitrogen's TOPO® cloning technology harnesses the activity of DNA topoisomerase I, which in nature functions both as a restriction enzyme and as a ligase. It cleaves and rejoins DNA during Vaccinia virus replication. Topoisomerase I specifically recognizes the pentameric sequence 5′-(C/T)CCTT-3′ and forms a covalent bond with the phosphate group of the 3′ thymidine. It then cleaves one DNA strand, enabling the DNA to unwind. The enzyme then religates the ends of the cleaved strand and releases itself from the DNA. To harness the re-ligating activity of topoisomerase I, TOPO® vectors are provided linearized with topoisomerase I covalently bound to each 3′ phosphate. This enables the vectors to efficiently ligate linear DNA fragments with compatible ends. The ligation is completed in only 5 min at room temperature.TOPO® PCR cloning only requires three steps: combine PCR product and a TOPO® cloning vector, incubate 5 min at room temperature, and transform E. coli (Figure 1). Cloning can be done 2 days faster by using TOPO® cloning kits with Mach1™-T1R competent cells compared to traditional restriction enzyme cloning (Table 2).Figure 1– TOPO® cloning protocol.We offer TOPO® cloning kits with Mach1™-T1R, the fastest-growing component cells on the market. With Mach1™-T1R competent cells, minipreps from an overnight colony are obtained in four hours—saving an entire day compared to cloning with most other strains.Table 2– TOPO® cloning kits with Mach1™-T1R competent cells save time.Versatility of TOPO® PCR cloningTOPO® cloning solutions are available for use with general subcloning, sequencing, or in vitro transcription; expression in E. coli or mammalian cells; or using the Gateway® system. The power of the technology enables the pcDNA™3.3-TOPO®–adapted mammalian expression vectors to yield tens of milligrams of recombinant protein expression in suspension-adapted mammalian cells.To request a sample today and learn more, visit www.invitrogen.com/toposample.FiguresReferencesRelatedDetails Vol. 46, No. 6 Follow us on social media for the latest updates Metrics Downloaded 699 times History Published online 25 April 2018 Published in print May 2009 Information© 2009 Future Science LtdPDF download

Multilevel and multidimensional Hadamard matrices

Multilevel Hadamard matrices (MHMs), whose entries are integers as opposed to the traditional restriction to {±1}, were introduced by Trinh, Fan, and Gabidulin in 2006 as a way to construct multilevel zero-correlation zone sequences, which have been studied for use in approximately synchronized code division multiple access systems. We answer the open question concerning the maximum number of distinct elements permissible in an order n MHM by proving the existence of an order n MHM with n elements of distinct absolute value for all n. We also define multidimensional MHMs and prove an analogous existence result.

Functional Oncogene Identification in Patient Leukemia Samples.

Approximately half of adult pre-B cell acute lymphocytic leukemias (pre-B ALL) harbor well-characterized chromosomal rearrangements known to affect prognosis and/ or treatment. In the 50% of cases with normal cytogenetics, pathogenesis is very poorly understood. Recent studies identified several somatic gene mutations in pre-B ALL samples. However, the functional significance of nearly all of these mutations remains unclear. We developed an approach to efficiently identify functionally-important, cryptic alterations directly from patient samples by establishing retroviral cDNA libraries. Streamlined methods for cDNA cloning and retroviral transduction have made it both feasible and cost-efficient to construct patient-specific libraries for oncogene identification. In our system (Figure), patient-specific retroviral cDNA libraries are used to infect the IL- 3-dependent B cell line, BaF3. Transformation by oncogenic kinases, transcription factors and other oncogenes results in IL-3 independent survival, allowing for the isolation of individual transformed clones and sequencing of integrated cDNA. cDNA is isolated from pre-B ALL samples and cloned into a retroviral backbone. Construction using clonase enzymes, rather than traditional restriction enzymes, maximizes efficiency and establishes representational cDNA libraries. BaF3 cells are infected with retroviral cDNA libraries then IL-3 is withdrawn. Viable, IL-3 independent clones are isolated and integrated cDNA are sequenced. If multiple cDNAs integrate into the same IL-3-independent clone, retrovirus is repackaged within that clone then applied at low titer to a new batch of IL-3- dependent BaF3 cells to identify the specific cDNA that confers IL-3 independence. With this approach, cryptic mutations, translocations and other rearrangements with functional significance can be identified. In addition, weaker oncogenes that require coordinated effects from other alterations can be elucidated by increasing the multiplicity of infection. To pilot the system, we assembled libraries using BCR-ABL-expressing K562 cells and peripheral blood lymphocytes from healthy donors. Restriction fragment analysis confirmed the presence of broadly heterogeneous cDNAs within the libraries. IL-3 independent cells were recovered after infection with the K562 libraries but not after infection with the healthy donor lymphocyte libraries. The IL-3 independent cells from the K562 library-infected population were confirmed to contain integrated BCR-ABL by PCR. We are currently characterizing transforming oncogenes recovered from pre-B ALL specimens with normal cytogenetics. Future efforts will characterize the biologic activity of these novel oncogenes and define the molecular epidemiology, as well as clinical features, associated with these alterations. In conclusion, large-scale sequencing efforts are underway at many centers to identify somatic mutations in tumor samples. Our functional approach offers an efficient alternative, focused on phenotypically meaningful, and therefore potentially drugabble, alterations. This approach could be easily applied to other tumor types, using either BaF3 or other cell lines that undergo oncogene-mediated transformation. [Display omitted]

Robust H∞ control based on fuzzy hyperbolic model with time-delay

This paper investigates the problems of robust stabilization and robust H ∞ control for the fuzzy hyperbolic model (FHM) with time-varying delay. A kind of novel Lyapunov–Krasovskii functional is constructed. By using the descriptor model transformation and free-weighting matrix technique, new delay-dependent sufficient conditions for the existence of desired FHM controllers are developed in terms of linear matrix inequality (LMI). The designed controllers not only ensure that the closed-loop system is asymptotically stable, but also satisfy a prescribed H ∞ performance level. The matrices of the Lyapunov–Krasovskii functional are not required to be diagonally positive-definite. In addition, the traditional restriction that the derivative of the time-varying delay is strictly smaller than one is not needed. Therefore, the obtained results are less conservative. Finally, a simulation example is provided to show the effectiveness of the proposed methods.

On eigenproblem solution of damped vibrations associated with gyroscopic moments

A new efficient approach is presented for solving the quadratic eigenvalue problem of weakly, nonproportionally damped vibration systems. In the analysis of these systems, gyroscopic moments and external damping are both considered. Traditional restriction of symmetry of inertia, damping and stiffness matrices is slightly relaxed. A second-order perturbation theory is developed such that the perturbed solution is based on the eigensolution of an unperturbed subproblem. This subproblem considers the unperturbed system in two different forms: (i) a conservative, gyroscopic part of an original problem, or (ii) a nonconservative, gyroscopic part of an original problem that is proportionally damped. To cope with asymmetry of the system matrices, a Duncan's like state formulation is used to bring these matrices into a suitable form for perturbations. Two numerical examples are introduced for explaining the detailed implementation of the presented approach. Additionally, a practical problem of rotor supported by two tilting pad-bearings is investigated. The eigensolutions obtained by the current approach match, to a great extent, other solutions obtained by time-consuming exact methods. The investigation procedure given here gives a framework to handle vibration problems of weakly nonproportional damping and/or weakly asymmetric inertia, damping and stiffness matrices.

Genotyping of Hepatitis B Virus – Genotypes A to G by Multiplex Polymerase Chain Reaction

Objectives: Eight genotypes (A–H) of hepatitis B virus (HBV) are known with variations in nucleotide sequences greater than 8%. Several recent publications found that the clinical course and outcome of antiviral therapy depended on the genotype of the infecting HBV strain. Large epidemiological studies will require the availability of a system which is rapid, reliable and can be performed on a large number of samples. Methods: To establish a simple and accurate genotyping method, the study collected 369 HBV complete genomic sequences from the GenBank database. Type-specific primers were also designed that separated HBV genotypes A to G by multiplex polymerase chain reaction. Results: By comparison with the traditional restriction fragment length polymorphism method, over 93% of 441 samples were accurately genotyped by current assay, with a higher detection rate and sensitivity to detect mixed HBV infections. Conclusions: This methodology can be applied only to areas prevalent with HBV genotypes A to G. However, it provides an efficient alternative for clinical diagnosis and large-scale studies.

Influence of simulated microwave disinfection on complete denture base adaptation using different flask closure methods

Denture microwave disinfection may result in dimensional changes that may distort the acrylic resin base, causing discomfort to the patient. The purpose of this study was to determine the effect of simulated microwave disinfection on maxillary denture base adaptation using 2 different flask closure methods. Forty stone cast-wax base sets were prepared for flasking by the traditional flask closure (TFC) and Restriction System flask closure (RSFC) methods (n=20). The RSFC consists of 2 iron plates to hold the flask during definitive flask closure, maintaining the flask in a closed position after release of pressure. Acrylic resin (Classico) was prepared according to the manufacturer's instructions. After polymerization in water at 74 degrees C for 9 hours, the bases were removed following flask cooling and submitted to conventional finishing with abrasive stones and pumice slurry. Ten bases for each TFC or RSFC method (n=10) were submitted to simulated disinfection (SD) in 150 mL distilled water in a microwave oven at 650 W for 3 minutes; control bases for each TFC or RSFC method (n=10) were not disinfected (ND). Three transverse cuts were made through each stone cast-resin base set, corresponding to the distal of canines, mesial of first molars, and posterior palatal region. Measurements were made in the bases using an optical micrometer at 5 points for each cut to determine adaptation: left and right marginal limits of the flanges, left and right ridge crests, and midline. Collected data were submitted to 3-way ANOVA followed by the Tukey HSD test (alpha=.05). Dimension gap values (mm) for ND denture bases prepared by the RSFC method were significantly lower (0.16 +/- 0.05) when compared to the TFC method (0.21 +/- 0.05) (P<.027). Simulated disinfection statistically improved the base adaptation in bases prepared by the TFC method (0.17 +/- 0.03), compared to the ND bases (0.21 +/- 0.05) (P<.027). Simulated disinfection statistically significantly improved base adaptation (P<.0001) only in the distal of canines (ND=0.13 +/- 0.01; SD=0.11 +/- 0.03) and the posterior palatal region (ND=0.25 +/- 0.04; SD=0.21 +/- 0.01) when bases were prepared by the TFC method. Simulated disinfection by microwave energy improved denture base adaptation when the TFC method was used, but did not statistically alter base adaptation for the RSFC method.

Spatial dispersion and energy in strong chiral medium

Since the discovery of backward-wave materials, people have tried to realize a strong chiral medium, which is traditionally thought impossible mainly for the reason of energy and spatial dispersion.We compare the two most popular descriptions of a chiral medium. After analyzing several possible reasons for the traditional restriction, we show that a strong chirality parameter leads to positive energy without any frequency-band limitation in the weak spatial dispersion. Moreover, strong chirality does not result in a strong spatial dispersion, which occurs only around the traditional limit point. For strong spatial dispersion where higher-order terms of spatial dispersion need to be considered, the energy conservation is also valid. Finally, we show that realization of strong chirality requires the conjugated type of spatial dispersion.

Identificación rápida de micobacterias no tuberculosas mediante análisis de patrones de restricción

The frequency of diseases caused by non tuberculous mycobacteria has increased in the last years. Their clinical diagnosis is difficult, mainly in immunocompromised patients. The identification of these mycobacteria by traditional methods is based on phenotypic characteristics and the results are obtained two to four weeks after their isolation in primary cultures. To report a new identification method for non tuberculous mycobacteria. The restriction pattern analysis method was implemented. It is based on the amplification, using polymerase chain reaction (PCR), of a polymorphic region of 440 base pairs that codifies Hsp65 protein, followed by a digestion with BstE II and Hae III restriction enzymes. The results were compared with patterns established for each strain. Sixty four strains of mycobacteria obtained from clinical samples and seven reference mycobacteria, were identified using the traditional methods and restriction pattern analysis. The latter method identified the same strain as the former in 87.5% of cases. In the remainder 12.5% of cases there was no agreement between both methods. In these, the sequencing of a fragment of a gene that codifies 16S ribosomal RNA, confirmed the correct identification by restriction patterns. Restriction pattern analysis is a rapid identification method for non tuberculous mycobacterial strains.

Detection of the Bcl I polymorphism of the glucocorticoid receptor gene by single-tube allele-specific polymerase chain reaction

The Bcl I polymorphism of the glucocorticoid receptor gene, recently identified as an intronic C to G change 646 nucleotides downstream of exon 2, has been associated with increased sensitivity to glucocorticoids and its potential relevance in metabolic disturbances and in various disorders has been extensively investigated. In the present study, we designed a single-tube allele-specific polymerase chain reaction for genotyping this polymorphism in peripheral blood DNA samples. When the Bcl I polymorphism was detected with this novel method in a cohort of 247 healthy subjects, the observed genotype distribution matched the Hardy–Weinberg equilibrium (100 subjects homozygous for the wild-type, 124 heterozygous and 23 homozygous for the mutant allele). In 50 randomly selected subjects the Bcl I polymorphism was also determined using a traditional restriction fragment length polymorphism technique and DNA sequencing, and the results showed 100% coincidence with those obtained by our novel method. The method proved to be more rapid and less labour-intensive compared to currently used techniques, and it avoided the use of extensive instrumentals. We assume that this novel method may have a broad utility in clinical and molecular epidemiological studies aimed to elucidate the impact of the Bcl I polymorphism of the glucocorticoid receptor gene either on metabolic disturbances, or various disorders, including cancer treatment and hormone substitution therapies.

The Interleukin-1β Gene Is Transcribed from a Poised Promoter Architecture in Monocytes

Cytokine transcription is usually regulated by transcription factor binding and chromatin remodeling following an inducing signal. By contrast, these data showed the interleukin (IL)-1beta promoter assembles into a "poised" structure, as evidenced by nuclease accessibility and loss of core histones immediately surrounding the transcription start site. Strikingly, these properties do not change upon transcriptional activation by lipopolysaccharide. Furthermore, association of two key transcriptional activators, PU.1 and C/EBPbeta, is robust pre- and post-stimulation indicating the IL-1beta promoter is packaged into a nontranscribed but poised promoter architecture in cells capable of rapidly inducing IL-1beta. Monocyte stimulation causes recruitment of a third factor, IRF-4, to the IL-1beta enhancer. PU.1 phosphorylation at a CK2 kinase consensus element is required for this recruitment. We showed that CK2 phosphorylates PU.1, CK2 inhibitors abrogate IL-1beta induction, and CK2 inducibly associates with the IL-1beta enhancer. Taken together, these data indicate a novel two-step mechanism for IL-1beta transcription: 1) formation of a poised chromatin architecture, and 2) phosphorylation of an enhancer-bound factor that recruits other activators. We propose that this poised structure may generally characterize rapidly activated genes.

Population Trends and Problems of Public Health

The scope and emphasis of a public health program are necessarily influenced by the changing characteristics of the population it serves. The rate of population growth affects long-range planning of community health and medical facilities. Alterations in age composition, internal migration of racial or industrial groups, changes in population density and urban-rural movement require current adaptation of the health program to solve the new problems thus created. Among the various characteristics of recent population trends, aging of the population is one of the most fundamental in its bearing on national health. The social and economic effects of an aging population have long been recognized. Dr. Louis I. Dublin appraised the problem of old age in some detail in 1926, when the provision of economic security for the aged was the dominant theme of contemporary discussion.1 The passage of the Social Security Act in 1935 represented the fruits of the efforts of this early period. Adjustment of national policy with respect to the health problems associated with aging of the population has been slower in development. Under the terms of the Social Security Act, a limited expansion of activities designed to promote the health of older adults—control of cancer and pneumonia, and industrial hygiene services—has been made possible in the cooperating States. However, the Act makes no provision for the solution of such fundamental problems as invalidity insurance and medical care of the aged. During the past five years, the health aspects of old age have received increasing attention in the discussions of public health administrators. It therefore seems appropriate to resurvey this general problem, and to consider, in particular, the nature of future trends in mortality, morbidity, and the receipt of medical care which may be expected solely as a result of changing age structure of the population.