Girls Lean Back Everywhere: The Law of Obscenity and the Assault on Genius

A Review of Girls Lean Back Everywhere: The Law of Obscenity and the Assault on Genius by Edward de Grazi

Is Private Health Care the Answer to the Health Problems of the World's Poor?

Background to the debate: The global burden of disease falls disproportionately upon the world's low-income countries, which are often struggling with weak health systems. Both the public and private sector deliver health care in these countries, but the appropriate role for each of these sectors in health system strengthening remains controversial. This debate examines whether the private sector should step up its involvement in the health systems of low-income countries

Good Health at Low Cost 25 years on: lessons for the future of health systems strengthening.

In 1985, the Rockefeller Foundation published Good health at low cost to discuss why some countries or regions achieve better health and social outcomes than do others at a similar level of income and to show the role of political will and socially progressive policies. 25 years on, the Good Health at Low Cost project revisited these places but looked anew at Bangladesh, Ethiopia, Kyrgyzstan, Thailand, and the Indian state of Tamil Nadu, which have all either achieved substantial improvements in health or access to services or implemented innovative health policies relative to their neighbours. A series of comparative case studies (2009-11) looked at how and why each region accomplished these changes. Attributes of success included good governance and political commitment, effective bureaucracies that preserve institutional memory and can learn from experience, and the ability to innovate and adapt to resource limitations. Furthermore, the capacity to respond to population needs and build resilience into health systems in the face of political unrest, economic crises, and natural disasters was important. Transport infrastructure, female empowerment, and education also played a part. Health systems are complex and no simple recipe exists for success. Yet in the countries and regions studied, progress has been assisted by institutional stability, with continuity of reforms despite political and economic turmoil, learning lessons from experience, seizing windows of opportunity, and ensuring sensitivity to context. These experiences show that improvements in health can still be achieved in countries with relatively few resources, though strategic investment is necessary to address new challenges such as complex chronic diseases and growing population expectations

Impact of bariatric surgery on hypertensive disorders in pregnancy: retrospective analysis of insurance claims data

Objective To determine whether women who had a delivery after bariatric surgery have lower rates of hypertensive disorders in pregnancy compared with women who had a delivery before bariatric surgery

Protein folding in the cell: challenges and progress.

It is hard to imagine a more extreme contrast than that between the dilute solutions used for in vitro studies of protein folding and the crowded, compartmentalized, sticky, spatially inhomogeneous interior of a cell. This review highlights recent research exploring protein folding in the cell with a focus on issues that are generally not relevant to in vitro studies of protein folding, such as macromolecular crowding, hindered diffusion, cotranslational folding, molecular chaperones, and evolutionary pressures. The technical obstacles that must be overcome to characterize protein folding in the cell are driving methodological advances, and we draw attention to several examples, such as fluorescence imaging of folding in cells and genetic screens for in-cell stability. DOI 10.1016DOI 10. /j.sbi.2010 Introduction Chris Anfinsen launched the field of protein folding by showing that ribonuclease (specifically, bovine pancreatic ribonuclease A) could refold to an active enzyme after reductive denaturation. Naturally, ribonuclease became emblematic of the fundamental tenet of protein folding -that the primary sequence of a protein specifies an energy landscape and a successful route to the native state at the global energy minimum. Yet ribonuclease folds in vivo during a complex journey through the secretory pathway of the cell. Notably, in its biological folding process, ribonuclease confronts milieux that are densely crowded with macromolecules; it samples the microenvironments of the ribosome tunnel, the translocon, and the ER lumen; it has the opportunity to fold from its N-terminus to C-terminus; and it is not left on its own, but instead is accompanied by lumenal chaperones that facilitate its folding and post-translational modification. As this journey abundantly illustrates, protein folding in the cell confronts many issues that are nonexistent in high dilution refolding experiments. It is thus not surprising that an increasing research effort is being applied to issues and processes involved in cellular protein folding. This review presents research in the expanding area of protein folding in the cell. We mainly confine our discussion to publications on cellular protein folding that have appeared in the last two years and to areas that have not been recently reviewed. We first describe a number of issues, such as macromolecular crowding and cotranslational folding, that arise when considering folding in the cell but are absent in vitro ( Macromolecular crowding A striking difference between most in vitro folding experiments and the cellular environment is the high concentration of macromolecules, which severely limits the cellular volume accessible to a polypeptide chain. Like many issues related to folding in the cell, determining the effects of crowding on folding presents major technical challenges to both computational and experimental studies; moreover, crowding is generally accompanied by other effects including altered diffusion and weak interactions. Macromolecular crowding also affects the viscosity of the cellular environment and solvent viscosity has been invoked as an important factor in determining folding rates and mechanisms (e.g., [9]). In a recent study, Dhar et al. used computational and experimental approaches to study the effects of a model crowder, Ficoll, on activity and folding of phosphoglycerate kinase (PGK), a 412-aa protein with two domains connected by a flexible hinge www.sciencedirect.com Current Opinion in Structural Biology 2011, 21:32-41 decreased the inter-domain separation. Also, the relaxation rate from temperature-jump unfolding experiments showed a maximum at 100 g/L Ficoll. The authors interpreted this result as arising from opposing effects of crowding and viscosity Hindered mobility and sticky neighbors Several recent studies show that the cellular environment affects macromolecular motion and provide insight into how. For example, two recent papers have used fluorescence recovery after photobleaching to monitor translational diffusion of GFP constructs in E. coli cells Vectorial synthesis and roles of mRNA and ribosomes in folding Newly synthesized polypeptide chains emerge from the ribosome vectorially, allowing their N-terminal portions to sample conformational space before the chain is completely synthesized. Additionally, the earliest environments encountered by a nascent chain are the ribosome tunnel and ribosome-associated chaperones. There have been excellent recent reviews on issues related to cotranslational folding including one in this issue A single domain stabilized by many long-range contacts is not expected to fold until the entire chain is complete, and recent studies of ribosome-bound nascent chains (RNCs) have confirmed this expectation for an SH3 domain by NMR [20] and GFP by observing chromophore maturation [21 ]. In fact, the NMR studies on SH3 RNCs reveal little or no compaction until the entire chain has exited the ribosomal tunnel [20]. By comparison, RNCs of the larger GFP may populate a more compact state before full translation [21 ]. The interrelationship of translation rate and folding has been discussed in a number of recent reviews [22][23][24][25]. Intuitively, slowing translation might allow more time for proper folding, and indeed in a recent study mutant ribosomes with reduced translation rates increased the soluble expression of eukaryotic proteins in E. coli [26 ]. The messenger RNA sequence can also affect the translation rate either through the use of rare codons [22,25,[27][28][29] or by RNA folding [29,30]. Either of these factors may be changed by synonymous mutations, where mRNA sequences are altered without affecting the encoded amino acid. Increasing the translation rate of the multidomain E. coli protein SufI by synonymously exchanging rare codon clusters for common codons was found to decrease cotranslational folding and the production of mature, folded protein [31 ]. Intriguingly, the mutation that leads to the deletion of F508 (DF508) in the cystic fibrosis transmembrane conductance regulator (CFTR), the most common mutation linked to cystic fibrosis, also changes the preceding codon for Ile507 [32 ]. Alteration of the local mRNA structure in the mutant retards translation and presumably impairs folding, increasing cotranslational ubiquitination and leading to protein degradation [32 ,33]. Restoration of the original Ile507 codon in the DF508 background significantly increases the amount of mature CFTR in the plasma membrane, demonstrating the potential impact of synonymous mutations on in vivo protein folding and maturation [32 ,33]. As described in recent reviews Molecular chaperones remodel the in vivo folding energy landscape The ability of molecular chaperones to interact with nascent or incompletely folded chains so as to favor successful folding and disfavor aggregation is well established. Yet the impact of chaperones on the folding mechanisms and stabilities of their clients is less clear, despite expanding literature on the functions and substrate-omes of several chaperones (e.g., Consider the case of arguably the best-studied chaperone, the E. coli chaperonin GroEL and its partner GroES. On Spatial organization, membranes and compartmentalization Cellular interiors are highly anisotropic with elaborate and physiologically critical architectures. This subcellular organization plays a major role in folding at several levels. For example, the native structures of membrane proteins are tuned to the diverse microenvironments and twodimensional character of membranes. Additionally, membranes are barriers, requiring proteins made on one side but destined to perform their functions extracytoplasmically to be translocated across the membrane, in most cases unfolded. For secretory proteins, passage through cellular compartments is highly choreographed with an assembly line of modifying enzymes, chaperones and transport mechanisms. Compartmentalization also opens up the possibility of chemical gradients, for example pH or oxidizing potential. Taken together, the spatial organization and compartmentalization of the cellular environment enable folding to occur in temporally optimized steps. Bacterial proteins destined for the periplasm or the cell exterior can be secreted across membranes either folded (e.g., by the Tat system) or unfolded (e.g., by the Sec system) Proteins that translocate through the Sec channel in bacteria and eukaryotes are greeted by an array of chaperones and modifying enzymes that alter the folding energy landscape. In addition, they move from an ATPrich, reducing environment to one that is ATP-poor and oxidizing as they enter either the bacterial periplasm, mitochondrial intermembrane space (IMS) or the eukaryotic ER. We refer interested readers to recent reviews of protein folding in the ER Exposure of nascent chains to the oxidizing environment of the periplasm or ER lumen enables step-wise disulfide bond formation, fixing the topology of secretory proteins. Not surprisingly, the timing and specificity of disulfide bond formation is integral to their in vivo folding, and this issue has been widely studied in eukaryotic proteins Many proteins that fold in the ER lumen are large with complicated topologies including multiple domains, disulfides, and glyosylation sites, which play roles in specialized in vivo folding mechanisms. A recent study on the influenza membrane glycoprotein neuraminidase Promising new methods for the study of in vivo protein folding The Holy Grail in studies of protein folding in the cell is to directly observe a protein of interest (POI) in intact cells and to characterize its folding, both thermodynamically and kinetically, in situ. Not surprisingly, this has proven exceedingly difficult. Several years ago, Ghaemmaghami and Oas took advantage of E. coli's urea tolerance to perform in-cell urea titrations of the l repressor headpiece utilizing a novel hydrogen exchange/ mass spectrometry method to assess stability Exciting recent work from the Gruebele lab combines temperature-jump perturbation methods with fast relaxation imaging (FReI) to interrogate the in vivo folding landscape of a POI, here a temperature-sensitive variant of phosphoglycerate kinase (tsPGK) In-cell NMR, recently reviewed by Pielak et al. [70] and by Ito and Selenko [71], is a potentially powerful approach to study proteins in vivo and to gain insight into their stability and folding mechanisms. Unfortunately, many folded proteins fail to show measurable NMR spectra in the cellular environment, most likely because of hindered rotational diffusion. Despite the inherent obstacles, the Shirakawa group has had impressive success applying NMR to small proteins in eukaryotic cells Clever use of split reporters, in which folding of the POI is coupled to successful binding and folding of two pieces of 36 Folding and Binding Figure 2 Monitoring protein folding kinetics in a living cell using fast relaxation imaging (FReI) These approaches to in-cell stability use a single flanking reporter and are clearly powerful as selections for enhanced stability. However, they are end-point assays based on the proteolytic lability of the fusion construct when the POI is unstable and therefore cannot readily yield an estimate of folding free energy. A related in vivo screen flanks the POI between a DNA-binding domain and a transcriptional activation domai

The 'Antiretrovirals, Sexual Transmission Risk and Attitudes' (ASTRA) study. Design, methods and participant characteristics.

Life expectancy for people diagnosed with HIV has improved dramatically however the number of new infections in the UK remains high. Understanding patterns of sexual behaviour among people living with diagnosed HIV, and the factors associated with having condom-less sex, is important for informing HIV prevention strategies and clinical care. In addition, in view of the current interest in a policy of early antiretroviral treatment (ART) for all people diagnosed with HIV in the UK, it is of particular importance to assess whether ART use is associated with increased levels of condom-less sex. In this context the ASTRA study was designed to investigate current sexual activity, and attitudes to HIV transmission risk, in a large unselected sample of HIV-infected patients under care in the UK. The study also gathered background information on demographic, socio-economic, lifestyle and disease-related characteristics, and physical and psychological symptoms, in order to identify other key factors impacting on HIV patients and the behaviours which underpin transmission. In this paper we describe the study rationale, design, methods, response rate and the demographic characteristics of the participants. People diagnosed with HIV infection attending 8 UK HIV out-patient clinics in 2011-2012 were invited to participate in the study. Those who agreed to participate completed a confidential, self-administered pen-and-paper questionnaire, and their latest CD4 count and viral load test results were recorded. During the study period, 5112 eligible patients were invited to take part in the study and 3258 completed questionnaires were obtained, representing a response rate of 64% of eligible patients. The study includes 2248 men who have sex with men (MSM), 373 heterosexual men and 637 women. Future results from ASTRA will be a key resource for understanding HIV transmission within the UK, targeting prevention efforts, and informing clinical care of individuals living with HIV

Cognitive Fatigue, Sleep and Cortical Activity in Multiple Sclerosis Disease. A Behavioral, Polysomnographic and Functional Near-Infrared Spectroscopy Investigation

Patients with multiple sclerosis (MS) disease frequently experience fatigue as their most debilitating symptom. Fatigue in MS partially refers to a cognitive component, cognitive fatigue (CF), characterized by a faster and stronger than usual development of the subjective feeling of exhaustion that follows sustained cognitive demands. The feeling of CF might result from supplementary task-related brain activity following MS-related demyelination and neurodegeneration. Besides, CF in MS disease might also stem from disrupted sleep. The present study investigated the association between the triggering of CF, task-related brain activity and sleep features. In a counterbalance mixed design, 10 patients with MS and 11 healthy controls were exposed twice for 16 min to a CF-inducing dual working memory updating task (TloadDback) under low or high cognitive demands conditions, counterbalanced. Considering known inter-individual differences and potential cognitive deficits in MS, the maximal cognitive load of the task was individually adapted to each participant’s own upper limits. During the experimental sessions, cortical brain activity was measured using near-infrared spectroscopy (NIRS) during the CF-induction task, and in a resting state immediately before and after. Ambulatory polysomnography recordings were obtained on the nights preceding experimental sessions. When cognitive load was individually adapted to their processing capabilities, patients with MS exhibited similar than healthy controls levels of subjectively perceived CF, evolution of performance during the task, and brain activity patterns. Linear mixed models indicate a negative association between oxygenation level changes in the dorsolateral prefrontal cortex (DLPFC) and the triggering of subjective CF in patients with MS only. Longer total sleep time was also associated with higher CF in MS patients. These results suggest that controlling for cognitive load between individuals with and without MS results in a similar task-related development of subjective CF. Besides comparable performance and cortical brain activity between groups, mixed model analyses suggest a possible association between CF, DLPFC activity and sleep duration in MS disease

Transcriptional and phenotypic characterization of novel Spx-regulated genes in Streptococcus mutans

CAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL E NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOIn oral biofilms, two of the major environmental challenges encountered by the dental pathogen Streptococcus mutans are acid and oxidative stresses. Previously, we showed that the S. mutans transcriptional regulators SpxA1 and SpxA2 (formerly SpxA and SpxB, respectively) are involved in stress survival by activating the expression of classic oxidative stress genes such as dpr, nox, sodA and tpx. We reasoned that some of the uncharacterized genes under SpxA1/A2 control are potentially involved in oxidative stress management. Therefore, the goal of this study was to use Spx-regulated genes as a tool to identify novel oxidative stress genes in S. mutans. Quantitative real-time PCR was used to evaluate the responses of ten Spx-regulated genes during H2O2 stress in the parent and Delta spx strains. Transcription activation of the H2O2-induced genes (8 out of 10) was strongly dependent on SpxA1 and, to a lesser extent, SpxA2. In vitro transcription assays revealed that one or both Spx proteins directly regulate three of these genes. The gene encoding the FeoB ferrous permease was slightly repressed by H2O2 but constitutively induced in strains lacking SpxA1. Nine genes were selected for downstream mutational analysis but inactivation of smu127, encoding a subunit of the acetoin dehydrogenase was apparently lethal. In vitro and in vivo characterization of the viable mutants indicated that, in addition to the transcriptional activation of reducing and antioxidant pathways, Spx performs an important role in iron homeostasis by regulating the intracellular availability of free iron. In particular, inactivation of the genes encoding the Fe-S biogenesis SUF system and the previously characterized iron-binding protein Dpr resulted in impaired growth under different oxidative stress conditions, increased sensitivity to iron and lower infectivity in rats. These results serve as an entryway into the characterization of novel genes and pathways that allow S. mutans to cope with oxidative stress.In oral biofilms, two of the major environmental challenges encountered by the dental pathogen Streptococcus mutans are acid and oxidative stresses. Previously, we showed that the S. mutans transcriptional regulators SpxA1 and SpxA2 (formerly SpxA and SpxB, respectively) are involved in stress survival by activating the expression of classic oxidative stress genes such as dpr, nox, sodA and tpx. We reasoned that some of the uncharacterized genes under SpxA1/A2 control are potentially involved in oxidative stress management. Therefore, the goal of this study was to use Spx-regulated genes as a tool to identify novel oxidative stress genes in S. mutans. Quantitative real-time PCR was used to evaluate the responses of ten Spx-regulated genes during H2O2 stress in the parent and Delta spx strains. Transcription activation of the H2O2-induced genes (8 out of 10) was strongly dependent on SpxA1 and, to a lesser extent, SpxA2. In vitro transcription assays revealed that one or both Spx proteins directly regulate three of these genes. The gene encoding the FeoB ferrous permease was slightly repressed by H2O2 but constitutively induced in strains lacking SpxA1. Nine genes were selected for downstream mutational analysis but inactivation of smu127, encoding a subunit of the acetoin dehydrogenase was apparently lethal. In vitro and in vivo characterization of the viable mutants indicated that, in addition to the transcriptional activation of reducing and antioxidant pathways, Spx performs an important role in iron homeostasis by regulating the intracellular availability of free iron. In particular, inactivation of the genes encoding the Fe-S biogenesis SUF system and the previously characterized iron-binding protein Dpr resulted in impaired growth under different oxidative stress conditions, increased sensitivity to iron and lower infectivity in rats. These results serve as an entryway into the characterization of novel genes and pathways that allow S. mutans to cope with oxidative stress104CAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL E NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL E NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCAPES [6849-12-1]FAPESP [2012/032278-3, 2014/03816-4]6849-12-12012/032278-3; 2014/03816-

Provision of the progestogen-only pill by community pharmacies as bridging contraception for women receiving emergency contraception:the Bridge-it RCT

Funding This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 25, No. 27. See the NIHR Journals Library website for further project information.Peer reviewedPublisher PD