The Relationship between Poverty and Healthcare Seeking among Patients Hospitalized with Acute Febrile Illnesses in Chittagong,Bangladesh

Delays in seeking appropriate healthcare can increase the case fatality of acute febrile illnesses, and circuitous routes of care-seeking can have a catastrophic financial impact upon patients in low-income settings. To investigate the relationship between poverty and pre-hospital delays for patients with acute febrile illnesses, we recruited a cross-sectional, convenience sample of 527 acutely ill adults and children aged over 6 months, with a documented fever ≥38.0°C and symptoms of up to 14 days’ duration, presenting to a tertiary referral hospital in Chittagong, Bangladesh, over the course of one year from September 2011 to September 2012. Participants were classified according to the socioeconomic status of their households, defined by the Oxford Poverty and Human Development Initiative’s multidimensional poverty index (MPI). 51% of participants were classified as multidimensionally poor (MPI>0.33). Median time from onset of any symptoms to arrival at hospital was 22 hours longer for MPI poor adults compared to non-poor adults (123 vs. 101 hours) rising to a difference of 26 hours with adjustment in a multivariate regression model (95% confidence interval 7 to 46 hours; P = 0.009). There was no difference in delays for children from poor and non-poor households (97 vs. 119 hours; P = 0.394). Case fatality was 5.9% vs. 0.8% in poor and non-poor individuals respectively (P = 0.001)—5.1% vs. 0.0% for poor and non-poor adults (P = 0.010) and 6.4% vs. 1.8% for poor and non-poor children (P = 0.083). Deaths were attributed to central nervous system infection (11), malaria (3), urinary tract infection (2), gastrointestinal infection (1) and undifferentiated sepsis (1). Both poor and non-poor households relied predominantly upon the (often informal) private sector for medical advice before reaching the referral hospital, but MPI poor participants were less likely to have consulted a qualified doctor. Poor participants were more likely to attribute delays in decision-making and travel to a lack of money (P<0.001), and more likely to face catastrophic expenditure of more than 25% of monthly household income (P<0.001). We conclude that multidimensional poverty is associated with greater pre-hospital delays and expenditure in this setting. Closer links between health and development agendas could address these consequences of poverty and streamline access to adequate healthcare.     

Cytogenetic Characterization and AFLP-Based Genetic Linkage Mapping for the Butterfly Bicyclus anynana,Covering All 28 Karyotyped Chromosomes

Background

The chromosome characteristics of the butterfly Bicyclus anynana, have received little attention, despite the scientific importance of this species. This study presents the characterization of chromosomes in this species by means of cytogenetic analysis and linkage mapping.

Methodology/Principal Findings

Physical genomic features in the butterfly B. anynana were examined by karyotype analysis and construction of a linkage map. Lepidoptera possess a female heterogametic W-Z sex chromosome system. The WZ-bivalent in pachytene oocytes of B. anynana consists of an abnormally small, heterochromatic W-chromosome with the Z-chromosome wrapped around it. Accordingly, the W-body in interphase nuclei is much smaller than usual in Lepidoptera. This suggests an intermediate stage in the process of secondary loss of the W-chromosome to a ZZ/Z sex determination system. Two nucleoli are present in the pachytene stage associated with an autosome and the WZ-bivalent respectively. Chromosome counts confirmed a haploid number of n = 28. Linkage mapping had to take account of absence of crossing-over in females, and of our use of a full-sib crossing design. We developed a new method to determine and exclude the non-recombinant uninformative female inherited component in offspring. The linkage map was constructed using a novel approach that uses exclusively JOINMAP-software for Lepidoptera linkage mapping. This approach simplifies the mapping procedure, avoids over-estimation of mapping distance and increases the reliability of relative marker positions. A total of 347 AFLP markers, 9 microsatellites and one single-copy nuclear gene covered all 28 chromosomes, with a mapping distance of 1354 cM. Conserved synteny of Tpi on the Z-chromosome in Lepidoptera was confirmed for B. anynana. The results are discussed in relation to other mapping studies in Lepidoptera.

Conclusions/Significance

This study adds to the knowledge of chromosome structure and evolution of an intensively studied organism. On a broader scale it provides an insight in Lepidoptera sex chromosome evolution and it proposes a simpler and more reliable method of linkage mapping than used for Lepidoptera to date.     

The direct peroxisome proliferator-activated receptor target fasting-induced adipose factor (FIAF/PGAR/ANGPTL4) is present in blood plasma as a truncated protein that is increased by fenofibrate treatment

The fasting-induced adipose factor (FIAF, ANGPTL4, PGAR, HFARP) was previously identified as a novel adipocytokine that was up-regulated by fasting, by peroxisome proliferator-activated receptor agonists, and by hypoxia. To further characterize FIAF, we studied regulation of FIAF mRNA and protein in liver and adipose cell lines as well as in human and mouse plasma. Expression of FIAF mRNA was up-regulated by peroxisome proliferator-activated receptor alpha (PPARalpha) and PPARbeta/delta agonists in rat and human hepatoma cell lines and by PPARgamma and PPARbeta/delta agonists in mouse and human adipocytes. Transactivation, chromatin immunoprecipitation, and gel shift experiments identified a functional PPAR response element within intron 3 of the FIAF gene. At the protein level, in human and mouse blood plasma, FIAF was found to be present both as the native protein and in a truncated form. Differentiation of mouse 3T3-L1 adipocytes was associated with the production of truncated FIAF, whereas in human white adipose tissue and SGBS adipocytes, only native FIAF could be detected. Interestingly, truncated FIAF was produced by human liver. Treatment with fenofibrate, a potent PPARalpha agonist, markedly increased plasma levels of truncated FIAF, but not native FIAF, in humans. Levels of both truncated and native FIAF showed marked interindividual variation but were not associated with body mass index and were not influenced by prolonged semistarvation. Together, these data suggest that FIAF, similar to other adipocytokines such as adiponectin, may partially exert its function via a truncated form.     

Functional Analysis of the Gene Cluster Involved in Production of the Bacteriocin Circularin A by Clostridium beijerinckii ATCC 25752

A region of 12 kb flanking the structural gene of the cyclic antibacterial peptide circularin A of Clostridium beijerinckii ATCC 25752 was sequenced, and the putative proteins involved in the production and secretion of circularin A were identified. The genes are tightly organized in overlapping open reading frames. Heterologous expression of circularin A in Enterococcus faecalis was achieved, and five genes were identified as minimally required for bacteriocin production and secretion. Two of the putative proteins, CirB and CirC, are predicted to contain membrane-spanning domains, while CirD contains a highly conserved ATP-binding domain. Together with CirB and CirC, this ATP-binding protein is involved in the production of circularin A. The fifth gene, cirE, confers immunity towards circularin A when expressed in either Lactococcus lactis or E. faecalis and is needed in order to allow the bacteria to produce bacteriocin. Additional resistance against circularin A is conferred by the activity of the putative transporter consisting of CirB and CirD.     

Species-specific plant–soil feedbacks alter herbivore-induced gene expression and defense chemistry in Plantago lanceolata

Oecologia - Plants actively interact with antagonists and beneficial organisms occurring in the above- and belowground domains of terrestrial ecosystems. In the past decade, studies have focused on...     

Alterations in the Cerebellar (Phospho)Proteome of a Cyclic Guanosine Monophosphate (cGMP)-dependent Protein Kinase Knockout Mouse

The cyclic nucleotide cyclic guanosine monophosphate (cGMP) plays an important role in learning and memory, but its signaling mechanisms in the mammalian brain are not fully understood. Using mass-spectrometry-based proteomics, we evaluated how the cerebellum adapts its (phospho)proteome in a knockout mouse model of cGMP-dependent protein kinase type I (cGKI). Our data reveal that a small subset of proteins in the cerebellum (∼3% of the quantified proteins) became substantially differentially expressed in the absence of cGKI. More changes were observed at the phosphoproteome level, with hundreds of sites being differentially phosphorylated between wild-type and knockout cerebellum. Most of these phosphorylated sites do not represent known cGKI substrates. An integrative computational network analysis of the data indicated that the differentially expressed proteins and proteins harboring differentially phosphorylated sites largely belong to a tight network in the Purkinje cells of the cerebellum involving important cGMP/cAMP signaling nodes (e.g. PDE5 and PKARIIβ) and Ca²⁺ signaling (e.g. SERCA3). In this way, removal of cGKI could be linked to impaired cerebellar long-term depression at Purkinje cell synapses. In addition, we were able to identify a set of novel putative (phospho)proteins to be considered in this network. Overall, our data improve our understanding of cerebellar cGKI signaling and suggest novel players in cGKI-regulated synaptic plasticity.Knockout (KO)¹ mouse models represent powerful methods for studying the physiological relevance of a protein. However, to elucidate the effects of KO-induced perturbations on the entire system, systems-wide molecular characterization is needed, as, for instance, provided by (phospho)proteomics. Recent technological and methodological advancements now allow the mapping of protein expression, at least in cell cultures, close to completion (1–3). More challenging, proteomics is also increasingly used to attempt systems-wide proteome characterizations in tissue. This has led to semi-quantitative (4–6) and quantitative (7) reasonably comprehensive proteome data on selected tissues, in both humans and animal models. More recently, proteomics has also been applied for the in-depth profiling of perturbations in the proteome occurring in KO models. For instance, de Graaf et al. (8) used an in-depth proteomic approach to identify the proteins changed by DNA-damage-induced premature aging, using a KO mouse model lacking the excision repair cross-complementing group 1 gene. Another recent study used a mouse model lacking apolipoprotein E in order to identify biomarker candidates for coronary artery disease (9).Adaptation and/or perturbations in the proteome caused by a KO can lead to changes in protein expression, but, at least equally likely, also to rewiring of signaling networks, through changes in post-translational modifications, such as protein phosphorylation. The application of (phospho)proteomics technology on KO or knock-in models is therefore also extremely relevant, albeit even more challenging. Hilger et al. (10) combined proteomics and phosphoproteomics on a cell line in which a phosphatase had been knocked out. To perform such experiments in a more (disease) relevant context, we should invest in functional, tissue-based phosphoproteomics approaches. A few examples of such approaches have very recently been reported. Lundby et al. (11) globally assessed phosphorylation events downstream of systemic adrenergic stimulation in mouse cardiac tissue. We recently reported on the use of a cardiac delimited CaMKII inhibited knock-in mouse to probe for substrates using a focused kinase-inhibition directed approach (12). Moreover, a mouse model lacking nitric oxide synthase (13), as a system of interest for Alzheimer disease, was recently studied via (phospho)proteomics.Here we explored how mature state-of-the-art (phospho)proteomics technology could be used to monitor the adaptation at the proteome level of the mouse cerebellum in a mouse line deficient for cGMP-dependent protein kinase type I (cGKI, also known as PKG-I), a kinase that plays an important role in synaptic plasticity, motor learning, and other brain functions (14). The cGMP-dependent protein kinases are serine/threonine kinases that act as key mediators of nitric oxide (NO) signaling, as well as of the natriuretic peptide pathway (15). In mammals, cGKs are encoded by two different genes: prkg1 coding for cGKI, and prkg2 coding for cGKII (16). The prkg1 gene encodes two cGKI isoforms, cGKIα and cGKIβ (17), which differ in their N-terminal leucine zipper and auto-inhibitory domains. cGKI regulates cardiovascular functions such as smooth muscle and cardiac contractility (16); in the nervous system it modulates synaptic plasticity in the hippocampus (18) and cerebellum (19).In the mammalian brain, more than 250 protein kinases are expressed, but only a few of these kinases are currently known to contribute to learning and memory. In particular, cGKIα is highly expressed in cerebellar Purkinje cells (PCs) (20, 21). Long-term depression (LTD) is an activity-dependent reduction in the efficacy of synaptic transmission and occurs at the PC synapses. Both a pharmacological approach using enzyme inhibitors (22) and a conditional PC-specific cGKI-KO (23) showed that cGKI plays a role in cerebellar LTD. Several proteins have been identified in past years as cGKI substrates in vitro or in cultured cells (15), but only a small portion of these have been confirmed as cGKI substrates in vivo. Therefore, the understanding of cGKI signaling and function depends strongly on the identification of novel in vivo substrates and signaling partners. In this perspective, the currently described approach allows us to discover potentially novel cGKI signaling routes and substrates directly in relevant cerebellar tissue. Our study revealed that cGKI-KO led to differential expression in the cerebellum of a specific group of proteins, of which many were closely connected to cGMP-cGKI signaling. More changes were observed at the phosphoproteome level, with the regulation of phosphorylation of a few hundred proteins. In particular, we hypothesize that some of the down-regulated phosphoproteins, but certainly not all, may be putative substrates of cGKI.