Functional interaction between neurons and glia is an exciting field that has expanded tremendously during the past decade. Such partnership has multiple impacts on neuronal activity and survival. Indeed, numerous findings indicate that glial cells interact tightly with neurons in physiological as well as pathological situations. One typical feature of glial cells is their high expression level of gap junction protein subunits, named connexins (Cxs), thus the membrane channels they form may contribute to neuroglial interaction that impacts neuronal activity and survival. While the participation of gap junction channels in neuroglial interactions has been regularly reviewed in the past, the other channel function of Cxs, i.e., hemichannels located at the cell surface, has only recently received attention. Gap junction channels provide the basis for a unique direct cell-to-cell communication, whereas Cx hemichannels allow the exchange of ions and signaling molecules between the cytoplasm and the extracellular medium, thus supporting autocrine and paracrine communication through a process referred to as “gliotransmission,” as well as uptake and release of metabolites. More recently, another family of proteins, termed pannexins (Panxs), has been identified. These proteins share similar membrane topology but no sequence homology with Cxs. They form multimeric membrane channels with pharmacology somewhat overlapping with that of Cx hemichannels. Such duality has led to several controversies in the literature concerning the identification of the molecular channel constituents (Cxs versus Panxs) in glia. In the present review, we update and discuss the knowledge of Cx hemichannels and Panx channels in glia, their properties and pharmacology, as well as the understanding of their contribution to neuroglial interactions in brain health and disease. Xenopus oocytes (2+) concentration in horizontal cells (2+ (2+ concentrations (1–2 mM) induced cell permeabilization, due to Cx43 hemichannel opening, before loss of membrane integrity  proteins can only function as gap junction channels. Indeed, before the aggregation of Cxs at the junctional plaque and subsequent formation of gap junctions, hexameric rings of Cxs, termed connexons, were initially assumed to remain closed. An obvious reason for this occlusion was that, as gap junction channels are “poorly” selective for ions and permeable to low molecular weight molecules (<1 to 1.2 KDa), if once at the membrane connexons could open, the cell would lose its integrity or at least would have to spend substantial energy to maintain this energetically unfavorable condition. Such statements began to be challenged when evidence for “functional hemichannels,” a term proposed to substitute for plasma membrane connexons, were reported in the early 1990s. In these pioneering studies, Cx hemichannels were opened either by large depolarization in  oocytes  or by loal cells . Later oells , was identified  and demoFigure 1). Cx hemichannels and Panx1 channels mediate the release of ATP that activates P2 receptors, promoting rises in intracellular Ca2+ concentrations; gap junction channels mediate the intercellular transfer of second messenger propagated calcium signal  that limits this feature -gated channel closure. Moreover, it was shown that the extracellular entrance does not undergo evident large conformational changes with either voltage-gating mechanisms  channels and calcium homeostasis modulator 1 (CALHM1) ion channel] and their possible involvement should be proved or disproved when testing the permeability of Cx hemichannels or Panx channels in a particular cell type.In addition to selective ion channels or transporters, other channels can alter the permeability of the cell membrane under physiological and pathological conditions . The ori+, ATP, adenosine, inositol trisphosphate (IP3), and glutamate; 2+; Table 1 for Cx43 hemichannels and Panx1 channels). In addition, it has been demonstrated that Cx hemichannels are permeable to synthetic molecules [e.g., Lucifer yellow, ethidium, calcein, propidium, 5(6)-carboxyfluorescein, and 2-(4-amidinophenyl)-1H-indole-6-carboxamide (DAPI); for review, see 2+ and cyclic nucleotides, respectively  allow the release of precursor or signaling molecules [e.g., including NADectively , indicatectively .via Cx hemichannels are in part explained by variations in the number of hemichannels available at the cell surface. With regard to this, an increase in the number of surface Cx43 or Cx45 hemichannels explains the increase in membrane permeability induced by fibroblast growth factor-1 (FGF-1) in cells that express these proteins -mediated phosphorylation of the six subunits of Cx43 hemichannels reconstituted in lipid membranes reduces the pore permeability allowing permeation of the small hydrophilic solute ethylene glycol (Mr 62) but not sucrose  depending on the permeant cationic species  it is important to have a library of permeant molecules that allow real time determination of their disappearance or appearance. Transport via Cx43 hemichannels has been shown to be cooperative and saturable with parameters  keep non-junctional Cx hemichannels closed in the plasma membrane, until they become incorporated into gap junction channels where they open by a process called “loop gating” as a consequence of extracellular loop interactions. In general, the half-maximal effective concentration for Ca2+-related opening of Cx43 hemichannels is in the order of 100 μM extracellular Ca2+. The proposed mechanism of millimolar Ca2+ closure of Cx hemichannels involves an interaction of extracellular Ca2+ with a Ca2+ binding site that consists of a ring of aspartate residues at the extracellular vestibule of the hemichannel pore thereby closing the channel  added to the interstitial fluid of a hippocampal acute slice preparation (discussed further below under Section “Impact of Hemichannel-Mediated Gliotransmission on Synaptic Activity and Behavior”).cent dye . Similarcent dye . Lowerin channel . Such a 2+Role of intracellular Ca. Besides extracellular Ca2+, intracellular cytoplasmic Ca2+ also influences hemichannel function. In fact, it was found that Cx hemichannel opening triggered by a decrease of extracellular Ca2+ could be inhibited by ester-loading the cells with the Ca2+ buffer 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetra-acetic acid (BAPTA), added to dampen intracellular Ca2+ changes  and negative feedback at higher concentrations. Interestingly, IP3 receptor channels present in the endoplasmic reticulum (ER) are Ca2+ release channels that have a similar bell-shaped Ca2+ dependency  and lanthanum (La3+) block hemichannels without inhibiting gap junction channels or Panx1 channels, but these ions also inhibit other channels including maxi-anion channels . The first Cx mimetic peptides that were introduced in the 1990s were identical to specific domains on the extracellular loops (first or second extracellular loop) of the Cx protein. The sequences mimicked were located in domains thought to be involved in the docking of two hemichannels during formation of a full gap junction channel , these peptides offer promising possibilities to explore the role of astroglial Cx43 hemichannels in the complex environment of intact neural tissues, ex vivo or in vivo. Recent work has, for example, demonstrated that injection of L2 peptide in the rat amygdala strongly suppresses fear memory  have been demonstrated to form functional hemichannels , and at  healthy . However healthy , 2011, bed above . This glbove amino)-2-deoxyglucose (2-NBDG), a glucose fluorescent analog, demonstrating that reactive or inflamed astrocytes can take up more glucose via Cx43 hemichannels when their metabolic coupling through gap junction channels is reduced  or the protocols used to induce neuronal death, (ii) the mode  of Cx channel inhibition selected, (iii) the type of neuronal death or pathology investigated, and (iv) the time points considered. However, retrospectively the opposite roles attributed to Cx channels could be simply due to the fact that for a long time this question was solely taking into account the gap junctional function of Cxs. Indeed, in most cases, interpretation of the data was not considering the hemichannel and channel functions of Cxs and Panxs, respectively. Nevertheless, the development of compounds and transgenic animals that discriminate between Cx gap junction channels, Cx hemichannels and Panx channels (see above) has allowed identification of their respective contribution in various in vitro and ex vivo models of brain pathologies -induced neuronal excitotoxicity is reinforced by TNF-α + IL-1β treatment, while such a potentiating effect is not observed in cultures of neurons alone treated with the two cytokines (Modifications in the pattern of Cx and Panx expression in glia are associated with phenotypic changes observed during neuroinflammation and the “reactive gliosis” that is a common feature of most brain diseases see . Indeed,out mice . Furtherytokines . This obytokines  is impaiytokines . This reytokines , preventytokines .In fine these gliotransmitters originating from glial cells trigger neuronal damage and induce their death. Thus, such a cascade of Cx glial hemichannel and Panx channel activation could partially account for the progression of the neurodegenerative disease. These in vitro and ex vivo observations could be related to the changes in Cx43 expression observed in reactive astrocytes that contact amyloid deposit in a murine model of Alzheimer’s disease (Connexin hemichannels might also play a role in the pathological context of neurodegenerative diseases. Indeed, it is becoming increasingly clear that glial cells are critical players in the progressive neurodegeneration of Alzheimer’s disease . Among o disease  and in b disease .During the last decade our view and understanding of the structure , mutatioversus hemichannels, the other is to discriminate between Cx hemichannels and Panx channels. To achieve these goals it is essential to have available pharmacological and genetic tools with glial cell type, Cx type, and Panx type specificities (see In the present review, we have focused on a Cx/Panx-based channel function which 10 years ago was only sparsely considered in the central nervous system and in neuroglial interactions. Since then, the identification of external and intracellular signals that trigger the activation of hemichannels, the understanding of their biophysics and their regulation have provided a better understanding of the roles that glial Cxs and Panxs play in healthy and diseased brains. Indeed, as reported above, there is now strong evidence to state that Cx and Panx channels in glia have an impact on neuronal activity and survival. However, there are at least two aims that remain to be achieved in order to determine whether and how the contribution of glial Cxs and/or Panxs is causal or secondary in normal and pathological situations. One is to distinguish what is due to gap junction channels ties see . Throughties see  or antibties see  could beties see . But herties see  gives weties see . Also, tties see , giving The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The acquisition of complex transcriptional regulatory abilities and epigenetic machinery facilitated the transition of the ancestor of apicomplexans from a free-living organism to an obligate parasite. The ability to control sophisticated gene expression patterns enabled these ancient organisms to evolve several differentiated forms, invade multiple hosts and evade host immunity. How these abilities were acquired remains an outstanding question in protistan biology.Perkinsus marinus, and in ciliates, the nearest free-living organisms with complete genome sequences available.In this work, we study SET domain bearing genes that are implicated in mediating immune evasion, invasion and cytoadhesion pathways of modern apicomplexans, including malaria parasites. We provide the first conclusive evidence of a horizontal gene transfer of a Histone H4 Lysine 20 (H4K20) modifier, Set8, from an animal host to the ancestor of apicomplexans. Set8 is known to contribute to the coordinated expression of genes involved in immune evasion in modern apicomplexans. We also show the likely transfer of a H3K36 methyltransferase (Ashr3 from plants), possibly derived from algal endosymbionts. These transfers appear to date to the transition from free-living organisms to parasitism and coincide with the proposed horizontal acquisition of cytoadhesion domains, the O-glycosyltransferase that modifies these domains, and the primary family of transcription factors found in apicomplexan parasites. Notably, phylogenetic support for these conclusions is robust and the genes clearly are dissimilar to SET sequences found in the closely related parasite Animal and plant sources of epigenetic machinery provide new insights into the evolution of parasitism in apicomplexans. Along with the horizontal transfer of cytoadhesive domains, O-linked glycosylation and key transcription factors, the acquisition of SET domain methyltransferases marks a key transitional event in the evolution to parasitism in this important protozoan lineage. Toxoplasma gondii, Cryptosporidium spp. and Plasmodium falciparum, the most lethal of human malaria parasites [The evolutionary processes whereby free-living or symbiotic organisms made the transition to full-fledged obligate, intracellular parasites remain unclear. This is perhaps best exemplified by the case of the Apicomplexa, a group of ~5000 species that includes several major human disease-causing agents including arasites . PresumaPlasmodium lineage, and more specifically the expansion of this domain in parasites that infect primates [The newly acquired lifestyle complexity might also require the acquisition of new mechanisms to control gene expression. For example, more sophisticated transcriptional regulation and epigenetic machinery would enable the evolution of complex life cycles involving multiple hosts and stages, and facilitate developmental changes accompanying the transition to parasitism. This could involve either de novo innovations or the horizontal acquisition of transcriptional and epigenetic machinery from other eukaryotes. Examples of both mechanisms of innovation have been reported. For instance, we previously described the unusual and rapid evolution of the C-terminal domain of RNA polymerase II within the primates . This doIn contrast, it is now well established that the primary class of transcription factors in apicomplexan parasites (the ApiAP2 family) was acquired through an ancient horizontal gene transfer (HGT) event . It is ki) transfer from an algal endosymbiont (fatty acid synthases or ApiAP2 transcription factors) [ii) transfer from an animal host (domains involved in cytoadhesion and O-linked glycosylation) [In the case of the Apicomplexa, therefore, two major potential sources for HGT have been established: factors) ,8, or iiylation) ,7. Here,P. falciparum, and are involved in imprinting genes involved in invasion and immune evasion [Histone lysine methyltransferases, characterized by a SET domain, play a fundamental role in gene activation and epigenetic regulation across all eukaryotes . These d evasion -15. Amon evasion . In receP. falciparum genome was reported by Cui et al. [An extensive examination of SET domain containing proteins (and the corresponding demethylases) found in the i et al. . One of i et al. . This prHomo sapiens Set8 (PR-7) as the query, we searched for the closest sequences to the Set8 domain across all major eukaryotic phyla. In many organisms, no conclusive Set8 ortholog could be identified, and in all taxa other than animals and apicomplexans, sequences retrieved using human Set8 as query proved to be most similar to other Set families in reciprocal blast searches of the complete NCBI protein database, suggesting that they might not be bona fide Set8 orthologs. Nonetheless, to avoid erroneously excluding highly divergent sequences, the nearest blastp matches to human Set8 were included in our analyses. Our complete dataset therefore represented potential Set8 sequences from all major eukaryotic lineages for use in phylogenetic analyses.The histone methyltransferase Set8 of apicomplexan parasites was recently shown to display strong sequence similarity to the orthologous protein in animals , suggestPerkinsus marinus and ciliates [Tetrahymena proteins are found in this clade, for instance, and Tetrahymena is known to lack Set8 [A phylogenetic tree was derived from combined maximum-likelihood (ML) and Bayesian inference using Set8 orthologs across this broad sampling of eukaryotic organisms with the results shown in Figure ciliates , apicompack Set8 ; as noteP. marinus genome provided a particularly valuable resource for investigating whether the Set8 transfer likely occurred specifically in the ancestor of extant apicomplexans. Perkinsus marinus, a parasite of oysters, is the nearest known relative of dinoflagellates and therefore serves as a representative sister group of apicomplexans for our analysis. Because dinoflagellates are predominantly free-living, Perkinsus must have evolved its parasitic lifestyle independently of the apicomplexans. The nearest SET domain bearing sequence to human Set8 from the P. marinus genome (E ≤ 10-13) did not return a match to Set8 when used as a query in a reciprocal blastp search, and nested deeply within the clade of non-Set8 sequences in phylogenetic analyses  in other organisms closely related to the apicomplexans, namely dinoflagellates, colpodellids, and Chromera velia, a photosynthetic autotroph discovered recently. The same was true for a variety of other protists including kinetoplastids, Trichomonas and Giardia, as well as fungi (including budding yeast), and also certain amoebozoans including Acanthamoeba.In an expanded survey of eukaryotes, including partial genomic resources available to date, blast queries with human Set8 failed to identify any significant hits  lack Set8, this provides evidence of a similar horizontal transfer of animal Set8 into slime molds. Considering the vast evolutionary distances between animals, the Apicomplexa and slime molds, and the absence of Set8 from all other eukaryotic lineages, it appears almost certain that the latter two groups acquired Set8 independently via HGT from animals. The highly unlikely alternative would be direct descent of apicomplexan Set8 from a common ancestor with animals, followed by gene loss repeatedly and independently from all other eukaryotes except some cellular slime molds.As shown in Figure P. falciparum, T. annulata, T.gondii, C. parvum and B. bovis) were used in phylogenetic analyses with only the animal and Dictyostelium sequences. To promote more accurate recovery of phylogenetic associations within the Set8 clade, we discarded highly divergent, non-orthologous SET sequences from other eukaryotes that could produce phylogenetic artifacts within the Set8 clade.The finding that apicomplexan Set8 is likely of animal origin raises questions about the approximate time of the transfer event, and the source of the acquired sequence. As noted above, the time of the transfer appears to be after the divergence of apicomplexans from dinoflagellates and the colopodellids  since thTrichonella spiralis. The ML bootstrap support for this relationship is weak (32%) but the Bayesian posterior probability is strong (0.99). Importantly, this tree recovers the same relationship between nematodes and Apicomplexa as did the expanded analysis, indicating this topology is stable regardless of the taxa sampled.As shown in Figure Dictyostelium and the Apicomplexa branch with nematodes, but separate from each other. If the positions of the Dictyostelium and apicomplexan sequences were long-branch attraction artifacts, they would be expected to attract each other as the two most divergent branches of the Set8 tree. Moreover, neither is attracted to the base of the Set8 clade, or even to the base of the nematode clade, but rather to individual sub-clades within the nematodes. This contrasts with the clear artefactual rooting of the overall Set8 clade, wherein long-branch outgroups are attracted to the rapidly evolving sequence from the parasitic trematode Schistosoma mansoni. Thus, although statistical support for a specific relationship between apicomplexan and nematode sequences is not strong, there appears to be no basis for concluding it is a typical phylogenetic artefact. While the precise timing of the emergence of nematodes is a subject of debate, all estimates place their origin between 600– 1200 million years ago [Furthermore, it is telling that both the slime mold ears ago . Thus, nTetrahymena and Paramecium) also possess putative Set2 homologs, suggesting that the Set2 family could have been present in the alveolate lineage before the divergence of the Apicomplexa from ciliates.Similar to Set8, Set2 is a histone methyltransferase that contributes to the regulation of higher order chromatin assembly and the epigenetic control of gene expression. Set2 and its cognate demethylase JmjC1 work by adding and removing methyl groups from H3K36, respectively . With thAravind and colleagues have argued that a major family of transcription factors (ApiAP2) in Apicomplexa were laterally transferred from the algal endosymbiont harbored intracellularly by all members of this group of parasites . This noPlasmodium proteins similar to the Set2 subfamily, as well as to Set1 and Set3  groups with Ashr3, a related H3K36 methyltransferase from green plants. An initial broader analysis performed with putative Set2 sequences from all apicomplexans recovered the same topology, but with weaker support  is available for tree reconstruction; potentially this could lead to spurious phylogenetic associations. In this case, however, there is additional corroborating evidence outside the SET domain for a close relationship between PF3D7_1322100 and the plant Ashr3 family. In both cases, the SET domain is positioned at the C-terminal end of the protein. In contrast, Set2 orthologs from animal, yeast and green plant models have a SET domain in more N-terminal locations 1 aa is a) identifPlasmodium and plant Ashr3 are unlikely to be coincidental. Rather, they represent credible evidence of an orthologous relationship. The Ashr3 Set subfamily is not found in animals or fungi [The combination of this conserved PHD in comparable synteny with the SET domain, similar overall domain architecture, and strong phylogenetic support for SET domain monophyly between or fungi  and, basor fungi , indicatCryptosporidum contains a four-PHD SET protein that groups with PF3D7_1322100 and all related apicomplexan sequences in expanded phylogenetic analyses  were extant at the time the ancestor of apicomplexans acquired the Set8 ortholog. Whereas arthropods are known definitively as hosts for many apicomplexans today, apicomplexan parasitism of nematodes has not been documented (perhaps because parasites of nematodes have been under-studied). Although What is the functional consequence of this transfer? Given the adaptations of apicomplexans toward intracellular niches, the acquisition of a major epigenetic regulator that governs and faithfully affirms chromatin-silencing marks could have been a critical step by protozoans for parasitism in diverse hosts with increasingly complex immune systems. H4K20Me1, the epigenetic mark deposited on histones by canonical Set8, is known to participate in gene silencing, heterochromatin formation and mitotic regulation as well as DNA damage checkpoint repair -29. We pP. marinus, the nearest available relative of apicomplexans, consistent with a proposed HGT coincident to the Set8 transfer. The animal sources of the cytoadhesion domain and O-linked glycosylation HGTs, however, remain unidentified [The observation that extant malaria parasites are capable of internalizing and incorporating exogenous host DNA spontaneously in animal cells is consistent with a potential mechanism of transfer ,30. Of sentified ,31. AlthC. elegans) could have transferred Set8 to the immediate ancestor of social amoebae and slime molds. Set8 orthologs are found in additional colonial slime molds  but are missing from other amoebae. Thus, the most parsimonious solution is an animal-derived HGT specific to slime molds. Caenorhabditis elegans is known to feed on Dictyostelium, ingesting and dispersing Dictyostelium spores [Dictyostelium[Similarly, the ancestor of modern soil-based nematodes  host polymerases [Cryptosporidium, whereas JmjC1 is absent, raising the intriguing possibility that they did not act together initially.Notably, while the phylogenetic analyses described here suggest horizontal acquisition of both Set8 and a Set2 functional analog at or near the origin of the apicomplexan clade, there is significant flexibility in the requirements and functions of these proteins in various extant organisms. This mirrors and extends our earlier investigations on transcriptional machinery in ymerases . FurtherPlasmodium specifically, the presence of additional RNA polymerase II CTD heptapeptides in primate parasites, a presumed requirement for recruitment of ‘Set2’, suggests that a cooperative interaction between ‘Set2’ and JmjC1 could have co-evolved with expansion of the CTD in these organisms. Consistent with this hypothesis, Plasmodium species that parasitize rodents do not have an expanded CTD and genes encoding Set2 and JmjC1 are missing from their genomes. Moreover, the deletion of both Set2 and JmjC1 in rodent plasmodia suggests tight and specific association between these chromatin modifiers in Plasmodium; this association is consistent with a recent integration into the genome, rather than with ancient, conserved functions that would have been harder to lose. The absence of epigenetic memory at rodent parasite surface-exposed variant antigens [yir family in P. yoelii) suggests a functional consequence of this deletion.In antigens  , and various metazoan taxa including sponges, nematodes, trematodes, arthropods, trochozoans, echinoderms, birds, reptiles and primates. NCBI accession numbers for both query and retrieved sequences used for this and other phylogenetic analyses are included on the respective tree figures in this paper.The queries from canonical orthologs of Set8 from Comparative evolutionary analyses were carried out on all sequences using the following approach. Sequences were aligned using the program MUSCLE  and trimPlasmodium Set2 and JmjC1 sequences. In these cases, we sampled NCBI protein databases with exemplars from both human and budding yeast, as well as sequences from P. falciparum and T. gondii. If different proteins were recovered using the alternative search strategies, both were included in initial respective phylogenetic analyses. For global phylogenetic analyses of SET domains, exemplars from model organisms  were chosen to define established SET protein subfamilies. The ciliate and Perkinsus sequences included were the closest match to both Set2 exemplars from model eukaryotes and to putative ‘Set2’ homologs from the two apicomplexans. Because results from both search strategies were comparable, sequences were assembled into a single alignment for phylogenetic reconstruction. Since different closest matches tended to emerge in blastp searches using apicomplexan versus human and yeast JmjC1 queries, separate phylogenetic analyses were carried out on the two data sets. With human and yeast queries, apicomplexan JmjC1 tended to group with various green algal sequences rather than with Perkinsus or ciliates, their nearest alveolate relatives with complete and well-annotated genomes. In contrast, querying with apicomplexan sequences resulted in trees that grouped the apicomplexans and ciliates together. The latter were considered to be the most conservative estimates of JmjC1 sequence relationships and, therefore, were included in as our primary phylogenetic analyses (see results).Comparable analyses were carried to test the phylogenetic affinity of The authors declare no competing financial, political, personal, religious, ideological, academic, intellectual, commercial or any other interests related to the work described in this publication.SPK, JWS and KWD designed the study. SPK identified and assembled the datasets for Set2, Set8 and JmjC1 sequences. JWS conducted the maximum-likelihood and Bayesian inference analyses and generated the phylogenetic trees. SPK, JWS and KWD wrote the paper. All authors read and approved the final manuscript.Expanded phylogenetic analyses of Set domain containing proteins showing that all putative Set2 (Ashr3) homologs from apicomplexans are recovered as a monophyletic group.Click here for file


Data on cause-specific mortality, skilled birth attendance, and emergency obstetric care access are essential to plan maternity services. We present the distribution of India's 2001–2003 maternal mortality by cause and uptake of emergency obstetric care, in poorer and richer states.The Registrar General of India surveyed all deaths occurring in 2001–2003 in 1.1 million nationally representative homes. Field staff interviewed household members about events that preceded the death. Two physicians independently assigned a cause of death. Narratives for all maternal deaths were coded for variables on healthcare uptake. Distribution of number of maternal deaths, cause-specific mortality and uptake of healthcare indicators were compared for poorer and richer states. There were 10 041 all-cause deaths in women age 15–49 years, of which 1096 (11.1%) were maternal deaths. Based on 2004–2006 SRS national MMR estimates of 254 deaths per 100 000 live births, we estimated rural areas of poorer states had the highest MMR  compared to the lowest MMR in urban areas of richer states . We estimated 69 400 maternal deaths in India in 2005. Three-quarters of maternal deaths were clustered in rural areas of poorer states, although these regions have only half the estimated live births in India. Most maternal deaths were attributed to direct obstetric causes (82%). There was no difference in the major causes of maternal deaths between poorer and richer states. Two-thirds of women died seeking some form of healthcare, most seeking care in a critical medical condition. Rural areas of poorer states had proportionately lower access and utilization to healthcare services than the urban areas; however this rural-urban difference was not seen in richer states.Maternal mortality and poor access to healthcare is disproportionately higher in rural populations of the poorer states of India. India contributes one-fifth of the global burden of absolute maternal deaths; however, it has experienced an estimated 4.7% annual decline in maternal mortality ratio (MMR) Within India, there is marked variation in MMR and healthcare access between regions and in socioeconomic factors In this study, we report on the maternal deaths in India's Sample Registration System (SRS). The SRS, with verbal autopsy, was used to estimate the national and regional distribution of maternal death and uptake of obstetric service indicators among women who died while pregnant or postpartum.RHIME) to monitor a nationally representative sample of 1.1 million households in the SRS. For every death occurring in these households from 2001–2003, a trained, non-medical RGI surveyor interviewed a relative or close-acquaintance of the deceased to obtain the symptoms and events around the death using structured questions and a local language narrative guided by a specific symptom list. These records were converted into electronic records and emailed to two of 130 trained physicians who, independently and anonymously, assigned an underlying cause of death , using guidelines for the major causes of obstetric and other deaths The MDS is an on-going nationally representative survey organized by the Registrar General of India (RGI). It is designed to determine the causes of death and risk factors of death. The design, methodology, and preliminary findings of the MDS have been described elsewhere Was the deceased pregnant, 42 day post-abortion, or 42 days post-partum? or that had been assigned an ICD-10 O-code (obstetric causes) were translated into English from 14 languages. All ICD-10 codes were reviewed by a central consensus panel, as described elsewhere Maternal deaths: All verbal autopsies forms with an affirmative answer to any of three questions primary care provider is the initial care provided to the woman for routine delivery or therapeutic abortion; community consultation is seeking healthcare in the community from a doctor, midwife or pharmacist in response to the onset of a complication; planned place of birth/abortion is place that care is sought at onset of labour/abortion; health-facility refers to any centre with admission capacity; emergency obstetric care refers to community consultation and/or health-facility admission; and highest healthcare uptake refers to ranking of the healthcare sought at any time the complication arose - routine admission to health-facility/hospital for abortion or labour, emergency admission for complication, community consultation for complication, and no facility-based healthcare utilization at time of complication. Emergency obstetric care is a proscribed set of indicators of staff skills, equipment, and supplies available 24 hours a day The following terms define obstetric care indicators: We used the RGI categorization of Indian poorer states, which have high-fertility, and maternal and infant mortality . Richer states are the remaining states and territories of India All proportional estimates account for weighting for sampling probability. Variance estimations were calculated using Taylor series linearization for the survey subpopulation of maternal deaths We estimated the number of maternal deaths and distribution of MMR by region and cause-specific mortality using the proportionate distribution of the survey-weighted sample, 2004–2006 SRS maternal mortality ratio estimates, and the United Nations Population Division estimates of live births and deaths in India in 2005 The proportion of missing data were imputed using multiple imputation by chained equations, and the distribution of observed versus imputed datasets were compared All analyses were conducted using Stata/SE .Registration of Births and Deaths Act, 1969. Verbal consent was obtained in the first SRS sample frame. The new SRS sample obtains written consent at the baseline. Families are free to withdraw from the study. The study poses no or minimal risks to enrolled subjects. All personal identifiers present in the raw data are anonymized before analysis. The MDS study using this SRS data has been approved by the review boards of the Postgraduate Institute of Medical Education and Research, Chandigarh, India and St. Michael's Hospital, Toronto, Canada.SRS enrolment is on a voluntary basis, and its confidentiality and consent procedures are defined as part of the Of 122 291 deaths there were 10 041 all-cause deaths in women age 15–49 years. There were 1130 pregnancy-related deaths, of which 34 were excluded as they were either accidental or incidental, resulting in 1096 maternal deaths . The national MMR reported by the SRS in 2004–2006 is 254 deaths per 100 000 live births (95%CI 239–269). We estimated that rural areas of poorer states had the highest MMR  compared to the lowest MMR in urban areas of richer states . We estimated 69 400 maternal deaths in India in 2005. Three-quarters of maternal deaths were clustered in rural areas of poorer states , whereas these regions have only half the estimated live births in India (13.3 million births). The proportion of maternal deaths to all-cause deaths in women, 15–49 years, was three times higher in rural areas of poorer states (16.3%) compared to urban areas of richer states .Half the maternal deaths were in the age range of 20–29 years, with a median age of 26 years (interquartile range 21–32 years), with no significant difference noted between poorer and richer states. Adolescent women (≤18) represented 6.0% (95%CI 4.4–7.6) of maternal deaths, and adolescent maternal deaths were not significantly over-represented in a single religious group, or rural versus urban areas (data not shown). The proportion of non-literate women was significantly higher for maternal deaths in poorer versus richer states (72.3% versus 46.1%) .There were no differences in the major causes of maternal deaths between poorer and richer states, or between rural and urban areas (data not shown). Most maternal deaths were attributed to direct obstetric causes  .One-quarter of maternal death were due to obstetric hemorrhage (n = 296), with most deaths occurring in the intrapartum period (n = 258). One-quarter of maternal deaths were due to ‘other obstetric complications’ (n = 248), which included ill-defined cause of death in labour (n = 125) and the antenatal and postpartum period (n = 81). Fifteen per cent of maternal deaths were due to indirect causes (n = 177).Maternal deaths making up the remaining sample were pregnancy-related infection, abortion, hypertension, and anesthetic complications from obstetric surgery. Pregnancy-related infection (n = 184) included puerperal sepsis (n = 130), antepartum deaths of sepsis onset following prolonged rupture of membranes (n = 30), and obstetric tetanus in the postpartum period (n = 24). Of the 108 maternal deaths due to complication in early pregnancy, most were reported complications following spontaneous abortion (n = 64).Earlier reporting of the same dataset found somewhat similar distribution of maternal deaths For most maternal deaths, the complication leading to death arose at term (≥7 months gestation), and the timing of complication onset were evenly distributed between the periods of pregnancy, intrapartum, and postpartum (≥24 hours–6 weeks post-delivery). One-quarter of the postpartum deaths occurred within the first three days and half died within 7 days of delivery .The proportion of maternal deaths who had ≥1 antenatal visit was significantly lower in the poorer versus richer states (58.3% versus 73.1%), and 11.5% delivered by caesarean, with no significant difference found between poorer and richer states.Maternal deaths in poorer states were more likely to seek consultation in the community; whereas, maternal deaths in richer states were more likely to transport directly to a health-facility or be there already whilst receiving routine care . Health-About half of the maternal deaths occurred at home (49.7%) and 13.8% occurred in transit . A significantly higher proportion of maternal death occurred at home in poorer versus richer states (53.8% versus 37.2). One-quarter of women in the sample received no healthcare contact from a midwife or physician (25.8%), either in the home, community, or health-facility.We looked at distribution of highest healthcare uptake sought by women with direct obstetric deaths at the time that the obstetric complication arose . HighestOur a priori subpopulation analysis compared rural to urban areas of two routine obstetric service indicators in both poorer and richer states  was a significant point of access to care for most women in a critical medical state in both poorer and richer states.One-third of complications arose in pregnancy prior to the onset of labour. This was higher than we expected. Narrative review permitted us to identify the timing of the complication relative to the onset of labour and we were able to identify the antepartum precedents of intrapartum mortality, unlike previous studies We were also able to differentiate between planned and actual place of birth, in order to differentiate between those seeking care for routine services and those seeking care in a critical medical condition. Routine care plays an important role in prevention and early identification of complications leading to maternal death Our study is the first nationally representative study of maternal mortality in India. The strength of this study is its size, as maternal deaths are relatively rare, permitting comparison across regions of India. Previous studies have been limited by small sample size, non-representative sampling, and measurement bias that does not differentiate between primary care provider versus emergency consultation care, and planned versus actual place of birth . In the Nonetheless, there are certain limitations to this study. These data are 10 years old however they do provide a national baseline of the characteristics of maternal deaths and healthcare use by these women, prior to the introduction of two major national health policy changes: the National Rural Health Mission, and the conditional cash-transfer scheme called Janani Suraksha Yojana. Secondly, a recent review of verbal autopsy coding found that physician coding performed as well or better than various automated coding methods, using hospital-based deaths as a reference Misclassification of cause-specific mortality may have occurred in differentiating between the types of maternal deaths, given the absence of medical care received in this population, leading to an underestimation of certain causes that often rely on objective measures (e.g. blood pressure for hypertensive disorders). In a systematic review of vital statistics and surveys, Khan et al. estimated the proportion of South Asian maternal deaths due to hypertensive disorders to be 9% (2–35%), and our estimate of 7% is within this range Misclassification of healthcare services may have occurred if the respondent was unaware of the skilled birth attendant's credentials or the health-facilities capacity to provide obstetric care, thus access to care may be over estimated. However, we do not have reason to believe that this misclassification occurred differentially across India.Incomplete data occurred as the primary study was not designed specifically to capture the reported variables of interest. Our missing data analysis  presentsThe maternal mortality ratio and the proportion of maternal deaths to all-cause death was higher in the poorer versus the richer states. We hypothesize that this difference is not due to any single specific cause of maternal death; however, perhaps our study did not have the power to demonstrate a difference in cause-specific mortality distribution between poorer and richer states. Further areas of research should look at how cause-specific mortality profile changes with various levels of MMR - as increasing healthcare uptake leads to a decline in preventable maternal deaths  Janani Suraksha Yojana, India's 2005 conditional cash transfer program to promote routine safe deliveries (skilled birth attendance and facility-based deliveries), future research should evaluate the effectiveness of this program on reducing the number of maternal deaths Reduction in India's maternal mortality rate would make an important contribution to the worldwide reduction of maternal mortality. Our analysis presents the important local variations from global characteristics of maternal mortality as well as the substantial internal variations within India. For policy-makers, faced with constrained budgets but committed to India's goal of effectively addressing a relatively rare yet highly important health priority, these variations may provide some targets for intensifying or initiating maternal health interventions. The majority of maternal deaths took place after 7 months gestational age and in the immediate post-partum period, and women are often presenting at obstetric facilities only in very serious condition. Thus, one priority is to provide health education on the early recognition of potentially hazardous conditions as part of an enhanced antenatal care program. As well, reduction in avoidable maternal deaths in India will require skilled healthcare providers with the capacity to deliver service for not only routine delivery but emergency obstetric care including community consultation and emergency admission to a health-facility. Secondly, obstetric services themselves could be a target area for intervention, particularly for investments in infrastructure, staffing and training in the rural areas of poorer states. Finally, we note that there was no difference in many outcomes between the rural and urban areas of the richer states, suggesting that national health and development programs focusing on poorer states must be maintained and even intensified. From 1999–2010, the proportion of safe deliveries has increased annually at twice the rate in the rural areas of poorer compared to richer states (8% versus 4%) Data sharing statement: The data used in this study are the property of the Registrar General of India and the overall mortality results have been published in 2006. Application for data access can be made to the Office of the Registrar General of India.Web Appendix S1(PDF)Click here for additional data file.


IQGAP1, an effector of CDC42p GTPase, is a widely conserved, multifunctional protein that bundles F-actin through its N-terminus and binds microtubules through its C-terminus to modulate the cell architecture. It has emerged as a potential oncogene associated with diverse human cancers. Therefore, IQGAP1 has been heavily investigated; regardless, its precise cellular function remains unclear. Work from yeast suggests that IQGAP1 plays an important role in directed cell growth, which is a conserved feature crucial to morphogenesis, division axis, and body plan determination. New evidence suggests a conserved role for IQGAP1 in protein synthesis and membrane traffic, which may help to explain the diversity of its cellular functions. Membrane traffic mediates infections by intracellular pathogens and a range of degenerative human diseases arise from dysfunctions in intracellular traffic; thus, elucidating the mechanisms of cellular traffic will be important in order to understand the basis of a wide range of inherited and acquired human diseases. Recent evidence suggests that IQGAP1 plays its role in cell growth through regulating the conserved mTOR pathway. The mTOR signaling cascade has been implicated in membrane traffic and is activated in nearly all human cancers, but clinical response to the mTOR-specific inhibitor rapamycin has been disappointing. Thus, understanding the regulators of this pathway will be crucial in order to identify predictors of rapamycin sensitivity. In this review, I discuss emerging evidence that supports a potential role of IQGAP1 in regulating membrane traffic via regulating the mTOR pathway.


Seasonally breeding birds detect environmental signals, such as light, temperature, food availability, and presence of mates to time reproduction. Hypothalamic neurons integrate external and internal signals, and regulate reproduction by releasing neurohormones to the pituitary gland. The pituitary gland synthesizes and releases gonadotropins which in turn act on the gonads to stimulate gametogenesis and sex steroid secretion. Accordingly, how gonadotropin secretion is controlled by the hypothalamus is key to our understanding of the mechanisms of seasonal reproduction. A hypothalamic neuropeptide, gonadotropin-releasing hormone (GnRH), activates reproduction by stimulating gonadotropin synthesis and release. Another hypothalamic neuropeptide, gonadotropin-inhibitory hormone (GnIH), inhibits gonadotropin synthesis and release directly by acting on the pituitary gland or indirectly by decreasing the activity of GnRH neurons. Therefore, the next step to understand seasonal reproduction is to investigate how the activities of GnRH and GnIH neurons in the hypothalamus and their receptors in the pituitary gland are regulated by external and internal signals. It is possible that locally-produced triiodothyronine resulting from the action of type 2 iodothyronine deiodinase on thyroxine stimulates the release of gonadotropins, perhaps by action on GnRH neurons. The function of GnRH neurons is also regulated by transcription of the GnRH gene. Melatonin, a nocturnal hormone, stimulates the synthesis and release of GnIH and GnIH may therefore regulate a daily rhythm of gonadotropin secretion. GnIH may also temporally suppress gonadotropin secretion when environmental conditions are unfavorable. Environmental and social milieus fluctuate seasonally in the wild. Accordingly, complex interactions of various neuronal and hormonal systems need to be considered if we are to understand the mechanisms underlying seasonal reproduction. Seasonal changes in reproductive physiology and behavior of birds are under hypothalamic control of pituitary gonadotropin  secretion. Gonadotropins secreted from the anterior pituitary gland stimulate gametogenesis  and sex steroid  synthesis in the gonad. Gonadal steroids induce the development of secondary sexual characteristics and facilitate reproductive behavior. A hypothalamic neuropeptide, gonadotropin-releasing hormone (GnRH), activates reproduction by stimulating gonadotropin synthesis and release. Another hypothalamic neuropeptide, gonadotropin-inhibitory hormone (GnIH), inhibits gonadotropin synthesis and release directly by acting on the pituitary gland or indirectly by decreasing the activity of GnRH neurons. This review attempts to understand how GnRH and GnIH neuronal systems can integrate external and internal environmental information and regulate gonadotropin secretion to time seasonal reproduction in birds Figure .Birds time their breeding so that young are hatched when there is enough food for them to be raised Lack, . SeasonaIn birds living in mid and high latitudes the annual change in day length controls the timing of breeding; photoperiod is thus the predominant proximate factor for birds living in such regions Follett, . Other eThe majority of temperate-zone passeriformes undergo dynamic seasonal changes in their reproductive activities. Gonadal development occurs in spring in response to increasing day length (“photostimulation”). However, the gonads are maintained in a functional condition only for a short period, and they spontaneously regress after extended exposure to long day lengths (“absolute photorefractoriness”). After becoming photorefractory, exposure to short day lengths is required for many passeriforme birds to regain “photosensitivity” and thus allow for photostimulation again :16 h dark (16D)] were placed in continuous darkness. When birds were exposed to a single 8 h photophase (light period), an increase in LH occurred only if the photophase coincided with a time period 12–20 h after the subjective dawn  of the hypothalamus, and eyes. Each of these sites may contain a circadian clock  were cloned in the Japanese quail. qPer2 and qPer3 showed robust circadian oscillations in the eye and in the pineal gland, although qClock mRNA was expressed throughout the day. qPer2 mRNA was induced by light, whereas qClock or qPer3 were not . Plasma melatonin concentration is always higher at night in both diurnal and nocturnal animals. Melatonin is synthesized from serotonin by the action of N-acetyltransferase to N-acetylserotonin, and by hydroxyindole-O-methyltransferase (HIOMT) to melatonin. Rhythms of melatonin synthesis and release from the pineal gland have been shown in pigeons, house sparrows, quail, and chicken. Circadian rhythms in melatonin secretion from the cultured pineal gland can persist in constant condition and they can be entrained by 24 h light-dark cycles  . Captive populations of mountain white-crowned sparrows showed robust gonadal development following transfer to longs days (15L:9D), but low temperature (5°C) slowed down photoperiodically induced gonadal growth and subsequent regression, in both males and females. High temperature of 30°C tended to accelerate gonadal development and regression whereas gonadal development was intermediate in a group exposed to 20°C. Prior exposure to these temperature regimes while on short days (9L:15D) had no effect. However, there was no effect of temperature on photoperiodically induced rises in LH in either sex despite marked effects on gonadal growth  differs by one amino acid from mGnRH-I, in that glutamine is substituted for arginine at position 8. Specific genes encoding the same cGnRH-I peptide were identified by cDNA cloning in Galliformes   differs by three amino acids from mGnRH-I or cGnRH-I at positions 5, 7, and 8. Most of the phylogenetic analyses support the existence of three GnRH clades in gnathostomes, named GnRH1, GnRH2, and GnRH3  were made, and the histological localization of GnRHs was studied in chicken and quail  have been identified with distinct distributions and functions in vertebrates  using GnRH antisera have demonstrated cyclic changes in GnRH in songbirds in response to changing photoperiod. RIA revealed that hypothalamic GnRH content did not increase significantly during the first 6 weeks of photostimulation in female starlings in a laboratory environment. However, by 12 weeks after the onset of photostimulation, as birds became photorefractory, GnRH had decreased to levels significantly lower than those before photostimulation , can mimic the effects of long photoperiods  of action of thyroid hormones may lie within the central nervous system since in starlings and sparrows, prevention of photorefractoriness by thyroidectomy is associated with maintenance of high levels of GnRH1, typical of photosensitive birds  catalyzes the conversion of T4 and T3 to their inactive forms. Interestingly, the expression of DIO3 was down-regulated under long day conditions when DIO2 was up-regulated  was studied in adult male Japanese quail , was a cardioexcitatory molecule isolated from the ganglia of the venus clam Macrocallista nimbosa  activation in a dose-dependent manner, and that the ratio of GnRH/GnIH receptors was a significant factor. From these results they proposed that in avian species, sexual maturation is characterized by a change in GnIH/GnRH receptor ratio, resulting in a switch in pituitary sensitivity from inhibitory, involving GnIH, to stimulatory, involving GnRH. In turn, decreasing GnIH-R signaling, combined with increasing GnRH-R-III signaling, results in significant increases in CRE activation, possibly initiating gonadotropin synthesis  of the hypothalamus, GnIH-ir neuronal fibers were widely distributed in the diencephalic and mesencephalic regions in birds  in this review to understand a central mechanism regulating seasonal reproduction Figure . There aThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


It has been recognized that the sympathetic nervous system is abnormally activated in chronic heart failure, and leads to further worsening chronic heart failure. In the treatment of chronic heart failure many clinical studies have already suggested that the inhibition of the abnormal sympathetic hyperactivity by beta blockers is beneficial. It has been classically considered that abnormal sympathetic hyperactivity in chronic heart failure is caused by the enhancement of excitatory inputs including changes in peripheral baroreceptor and chemoreceptor reflexes and chemical mediators that control sympathetic outflow. Recently, the abnormalities in the central regulation of sympathetic nerve activity mediated by brain renin angiotensin system-oxidative stress axis and/or proinflammatory cytokines have been focused. Central renin angiotensin system, proinflammatory cytokines, and the interaction between them have been determined as the target of the sympathoinhibitory treatment in experimental animal models with chronic heart failure. In conclusion, we must recognize that chronic heart failure is a syndrome with an abnormal sympathoexcitation, which is caused by the abnormalities in the central regulation of sympathetic nerve activity. Sympathetic nervous system has a wide variety of cardiovascular actions, including heart rate acceleration, increase in cardiac contractility, reduction of venous capacitance, and constriction of resistance vessels , 2. It hActivation of sympathetic nervous system, reduction of the vagal activity, and the secretion of renin angiotensin-aldosterone axis are occurred in chronic heart failure with left ventricular systolic dysfunction , 2, 5 an There are several animal models with chronic heart failure, and those animal models may mimic the human condition with chronic heart failure closely . In spit1) receptors are found in the central nervous system and are expressed to a high degree in areas of the hypothalamus and medulla, which regulate sympathetic outflow [1 receptor in the paraventricular nucleus (PVN) of the hypothalamus in chronic heart failure with postmyocardial infarction [In the brain, renin angiotensin system is considered to be a main system of regulating sympathetic nervous system . In the  outflow , 19. Aldfarction . These p1 receptor-induced oxidative stress in the RVLM causes sympathoexcitation in the animal models with chronic heart failure [1 receptor blocker into the RVLM causes sympathoinhibition in experimental chronic heart failure [1 receptor protein, AT1 receptor mRNA, and angiotensin II levels are increased in the RVLM and nucleus tractus solitarii (NTS) in rabbits and rats with chronic heart failure [1 receptor and oxidative stress plays a critical role in the abnormal sympathoexcitation in chronic heart failure. Furthermore, the balance between angiotensin-converting enzyme (ACE) and its homolog ACE2 or between AT1 and angiotensin II type 2 receptor in the brain may be an important determinant of sympathoexcitation in chronic heart failure [1 receptor-induced oxidative stress in the brain, especially in the RVLM, might be a novel target of the therapy for chronic heart failure through the sympathoinhibition. As the mechanisms in which brain renin angiotensin system causes sympathoexcitation, brain oxidative stress has been focused. Brain renin angiotensin system is involved in the production of oxidative stress in the brain , 21–23.  failure , 28. Mic failure –18. AT1  failure . These p failure , 29, 30.κB) [κB-mediates cross talk between proinflammatory cytokines and brain renin angiotensin system in rats with chronic heart failure [Brain inflammatory mediators and the brain renin angiotensin system are both implicated in sympathoexcitation in experimental chronic heart failure , 32. RecκB) , or micrκB)  in the bκB) . There mκB) . In addiκB) . Moreove failure , 37. Int failure . CombineWe have also demonstrated other several mechanisms of abnormal sympathoexcitation in chronic heart failure. In the brain, nitric oxide (NO) causes sympathoinhibition , 48, andMany clinical studies have already and strongly suggested that chronic beta blocker therapy improves left ventricular performance and reverses left ventricular remodeling, reduces risk of hospitalization for heart failure, and improves survival of chronic heart failure –61. AmonCentral alpha2 receptor has been considered to be possible targets of treatment for chronic heart failure, because the excitation of the central alpha2 receptor causes sympathoinhibition , 65. In 1 receptors is present in brain regions outside of the blood-brain barrier where peripherally administered AT1 receptor blockers are able to access without considering the existence of the blood-brain barrier as well as inside of the blood-brain barrier [1 receptor blockers also act on the AT1 receptors within the brain, thereby reducing blood pressure in hypertensive rats [1 receptor blockers could cause beneficial sympathoinhibition via blockade of brain renin angiotensin system. Angiotensin II and aldosterone production enhances the release and inhibits the uptake of norepinephrine at nerve endings . ACE inh barrier . Recent ive rats , 73–77. 1 receptor and production of superoxide with upregulating NO synthase in the RVLM of the animal models with chronic heart failure [ Several studies in rabbits with pacing-induced heart failure have demonstrated that statins normalize abnormal sympathetic hyperactivity in experimental chronic heart failure –80. Prev failure , 80. We  failure –83. AlthAs the nonpharmacological therapy for chronic heart failure, exercise training is considered to have sympathoinhibitory benefit on chronic heart failure. Exercise intolerance is a characteristic of patients with chronic heart failure, and skeletal myopathy contributes to the limitation of functional capacity in chronic heart failure , 2, 9. A Recent novel topic in the therapy with sympathoinhibition is renal sympathetic denervation. Renal afferent nerves may also contribute to the blood pressure elevation according to the recent findings of the renal nerve ablation in patients with resistant hypertension –91. RenaIn chronic heart failure, it has been recognized that the abnormal sympathoexcitation occurs. In the treatment of chronic heart failure, the therapy with sympathoinhibition, such as beta blockers and/or exercise training, has already been considered to be important. We must recognize that chronic heart failure is a complex syndrome with sympathoexcitation and that the abnormal sympathoexcitation should be the target of the treatments for chronic heart failure. In this aspect, conservative pharmacological therapy is not sufficient, and the additive new and novel device therapy and/or nonpharmacological therapy are necessary.1 receptors, oxidative stress, and inflammatory pathway in the brain are novel sympathoinhibitory therapeutic targets for chronic heart failure ( The mechanisms in which the abnormal sympathoexcitation occurred in chronic heart failure have not been fully determined. Particularly, the central abnormalities need further examinations in clinical and basic research. It is interesting and important to consider that AT failure .


It is well established that nutritional habits are relevant in the prevention of Idiopathic Calcium Nephrolithiasis (ICN) and the Mediterranean diet is believed to be protective against nephrolithiasis and not only against cardiovascular events. A case-control study was performed to establish the nutritional habits of Italian ICN patients and the nutritional determinants of lithogenic risk in the considered population.2) and controls  were enrolled. A 3-day nutritional diary was analyzed by the software Dietosystem . The nutritional intake was also compared to the Italian nutritional guidelines. Urinary factors were analyzed from 24h urine collection and statistical analysis was performed by the SPSS software.Calcium stone formers  and a decreased excretion of K+  and citrate  in SF than CTR. Nutritional analysis found differences between SF and CTR: a higher caloric intake , a higher total protein intake  and a higher vegetable protein intake  were observed in SF than CTR. Moreover SF showed a higher intake of sodium, oxalate, complex carbohydrates, purines, arachidonic acid and a higher acid load. The comparison of SF nutritional intake to the Italian guidelines demonstrated a prevalence of hyperproteic, hyperlipidic and hypercaloric diet, a low calcium, potassium and fiber intake, and a high salt, phosphorus and oxalate intake. Water intake covered the recommended quantity (1.5-2 L/die) for healthy population, but not for SF (>2 L/die).Urinary data showed an increased excretion of CaIn conclusion, our study found different nutritional habits between SF and CTR and confirmed some dietetic errors respect to national dietetic recommendations. These errors could increase the risk to develop ICN in subjects with a lithogenic genetic background and confirm the usefulness to give nutritional advices to SF patients.


RFCA has been established as an effective and curative therapy for severely symptomatic PVC from the outflow tract in structurally normal hearts. However, it is unknown whether PVCs originating from the left ventricular septum, are effectively eliminated by RFCA. This study aimed to investigate electrophysiologic characteristics and effects of Radiofrequency catheter ablation (RFCA) for patients with symptomatic premature ventricular contraction (PVC) originating from the left ventricular septum without including fascicular PVCs.Characteristics of body surface electrocardiogram (ECG) and electrophysiologic recordings endocardiogram in a successful RFCA target were analyzed in 20 patients with symptomatic PVCs originating from the left ventricular septum. RFCA was performed using pace mapping and activation mapping.The QRS morphology of PVCs originating from the left ventricular septum is similar to that seen in fascicular tachycardia. Most of the PVCs originated from the left septum appears in the form of ventricular parasystole. The incidence of ventricular parasystole was 70%. Sustained ventricular tachycardia was not inducible by electrical stimulation and isoproterenol infusion in all 20 patients, ablation at the site recording the earliest Purkinje potential was not effective in all 20 patients, and Purkinje potentials were not identified at successful sites during point mapping. Sixteen patients were successful with RFCA using pace mapping and activation mapping, 3 failed, and 1 recurrent.Although the ECG characteristics of the PVCs arising from the left ventricular septum are similar to that seen in fascicular tachycardia, the electrophysiologic characteristics are different between the two types of PVCs. The distinguishing characteristic of the PVCs is that Purkinje potentials were not present at the site of successful ablation, suggesting a myocardial as opposed to fascicular substrate. RFCA is an effective curative therapy for symptomatic PVCs originating from the left ventricular septum (not from the left anterior and posterior fascicle). Isolated premature ventricular complexes (PVCs) are the most common arrhythmias observed in patients without structural heart disease . In receFrom September 2006 to May 2010, a total of 318 patients without structural heart disease were presented for catheter ablation for PVCs in our hospital. Only patients with idopthic PVCs from the left ventricular septum were enrolled in the present study. All patients were verified as having no structural heart disease, including coronary artery disease, valvular heart disease, congenital heart disease, left ventricle hypertrophy, and right ventricle abnormalities by routine biochemistry tests, X-ray, color echocardiography examination, exercise electrocardiogram testing, and/or cardiac catheterization with coronary angiography. Before RFCA, a 12-lead ECG was obtained, and 24 hours ambulatory ECG monitoring (Holter) was carried out at least once. The ECG was monitored for 24 hours just before catheter ablation.The selection criteria of patients were the following reasons: (1) frequent PVCs occurrence, the average PVC count ≥ 10000 times/24 h; (2) inability of the patient to tolerate PVCs or unsuccessful treatment with at least two antiarrhythmic drugs; (3) no structural heart disease; and (4) consent for the catheter ablation procedure.Twelve-lead electrocardiograms of the clinical arrhythmia were available for all patients with PVCs originating from the left ventricular septum. The analysis of ECG pattern was focusing on the following characteristics: (1) The QRS morphology of the PVC in all 12 leads, (2) the duration of QRS complex, (3) the axis deviation.Electrophysiologic study was performed after withdrawal of all anti-arrhythmic drugs for at least five half-lives. Under fluoroscopic guidance, two quadripolar electrode catheter  were advanced from the right femoral vein and positioned in the right ventricular apex and near the His bundle. One additional catheter was inserted via the internal jugular vein and positioned in the coronary sinus. A 12-lead surface electrocardiogram was monitored and recorded on a multichannel oscilloscopic recorder. Programmed electrical stimulation was performed from right ventricular apex at basic drive cycle lengths of 600, 500, and 430 msec, delivering a maximum of three extrastimuli. A 7F quadripolar deflectable catheter was advanced into the left ventricle using retrograde aortic approach via the right femoral artery for mapping and ablation . The mapping/ablation catheter has a 4 mm distal electrode, with interelectrode spacing of 2-5-2 mm. Pace mapping and endocardial activation mapping were performed. If the clinical arrhythmia did not occur spontaneously and was not induced in the baseline, intravenous isoproterenol (2-4 μg/min) was administered to induce arrhythmia.The target site of RFCA was determined by two steps:1) a careful search was firstly performed for Purkinje potentials preceding the onset of QRS during sinus rhythm along the left ventricular septum in all patients, and radiofrequency energy was delivered at sites demonstrating local endocardial recording with earliest presystolic Purkinje potentials during PVCs, or in sinus rhythm if there was insufficient ectopy to permit activation mapping; 2) next step is to obtain the complete or near complete pace mapping with the earliest local activation time if RFCA is not successful, and when the PVC occurred rarely during the electrophysiologic study, only pace mapping was performed, aiming to identify the site where pacing reproduced QRS morphology that is similar to the clinical PVC (≧ 11/12-lead concordance of major and minor deflections). After the target site was located, the energy of RFCA was delivered using maximum power of 40 W, maximum temperature of 60°C and impedance of 80-140 Ωin the temperature-controlled mode. If the PVC was terminated within 10 seconds or frequently PVCs and/or nonsustained ventricular tachycardia occurred during ablation at the target site, additional current was applied for another 60 to 180 seconds. Successful ablation was defined as complete elimination of spontaneous or inducible ventricular arrhythmias. Programmed electrical stimulation was repeated at 30 minutes after the last application of radiofrequency energy to confirm the absence of inducible ventricular arrhythmias before removing all catheters and sheaths. If PVC did not terminate within 10 s, the radiofrequency energy application was terminated and another target site was sought.Successful ablation was defined as the absence of spontaneous or induced clinical PVC and ventricular tachycardia, both with or without isoproterenol at the end of the procedure. Absence of PVC or ventricular tachycardia in ECG monitoring over 48 hours without anti-arrhythmia drugs. No recurrence of arrhythmia in the absence of anti-arrhythmic drug therapy during follow-up.After RFCA, all patients underwent 72-hour ECG monitoring. Holter was carried out one week after RFCA. Patients were not given any antiarrhythmic drugs after RFCA, and underwent color echocardiography and Holter examination three month after RFCA. ECG echocardiography and 24-hour ECG monitoring were performed whenever the patient had symptoms suggestive of recurrence of ventricular arrhythmias..PVC was considered to originate from the left ventricular septum, not from the left anterior and posterior fascicles, based on (1) the local endocardial recordings; (2) the characteristic of left ventricular septum location and motion (when viewed in the right and left anterior oblique fluoroscopic views after successful RFCA); (3) programmed electrical stimulation could not induce sustained ventricular tachycardia; (4) unsuccessful ablation at Purkinje potential site, and (5) purkinje potentials were not identified at successful sites during point mapping.Ethical approval was obtained from the Ethics Committee of the Second Affiliated Hospital of Wenzhou Medical College, and all patients gave informed consent before participation in the study.A total of 20 patients  were enrolled in the present study. Among patients with idiopathic ventricular ectopy, the incidence of left septal ventricular arrhythmias was 6.29%  by pace mapping and activation mapping, not by searching for Purkinje potential in the left ventricular septum, and found that most cases with the PVCs arising from the left ventricular septum behaved with parasystolic activity.In this study, we found that PVCs originating from the left ventricular septum have distinctive ECG characteristics. First, the electrocardiographic patterns are different at the different sites of origin of left septal PVC. For PVCs originating from left anterosuperior septum, their QRS complex morphology were qR or qRs in leads II, III, aVF, Rs (R/s > 1) in leads V5~V6, rs(S) in leadsI, aVL and Qr in lead aVR. For PVCs originating from left posteroinferior septum, their QRS complex morphology were rS in leads II, III, aVF, R(r)S (R/S < 1) in leads V5~V6, qR(s) in leadsI,aVL and qR in lead aVR. The QRS morphology of PVCs originating from the left ventricular septum is similar to that seen in fascicular tachycardia . Why areIn this study, ventricular parasystole was often observed, sustained ventricular tachycardia was not inducible by electrical stimulation and isoproterenol infusion in all 20 patients, ablation at the site recording the earliest Purkinje potential was not effective in all 20 patients, and Purkinje potentials were not identified at successful sites during point mapping. These suggest enhanced antomaticity, but not reentry as the most likely mechanism of PVCs originating from the left ventricular septum in the study ,14. The In this study, we demonstrated PVCs arising from the left ventricular anterosuperior and posteroinferior septum can be safely and effectively eliminated with catheter ablation techniques. The immediate ablation success rate was 85%, and the chronic success rate was 80% (16 of 20 patients) during a mean follow-up period of 20.2 months. One patients experienced significant recurrence of PVCs with associated symptoms. The overall success rate for ablation of PVCs originating from the left ventricular septum in the present study nearly corresponded with the results of previous reports for ablation of PVCs originating from RVOT and/or other sites of origin ,15,16. TFirst, the mechanism of the PVCs arising from the left ventricular septum is thought to be due to enhanced antomaticity, but not reentry, but this remains speculative in the limited clinical study. Second, although the PVCs may occur from the myocardium of the septum, instead of the Purkinje network, it is unknown that how distributed were these septal sites. To increase the accuracy of our study, our results need to be confirmed in a larger prospective randomized patient population.Although the ECG characteristics of the PVCs arising from the left ventricular septum are similar to that seen in fascicular tachycardia, the electrophysiologic characteristics are different between the two types of PVCs. Most cases with the PVCs arising from the left ventricular septum behaved with parasystolic activity. The results of the present study confirm the high success rate and safety of RFCA using conventional techniques in the management of PVCs originating from the left ventricular septum.The authors declare that they have no competing interests.LJF and LYC designed the whole study, LJF, LJ, LYC, JKT, ZND, LJX, ZWW, YPL, and TJF performed the experiment, LJF and LYC wrote the paper. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2261/11/27/prepub


Menopause is characterized by depletion of ovarian follicles, a reduction of ovarian hormones to castrate levels and elevated levels of serum gonadotropins from the anterior pituitary gland. Although this process has significant repercussions throughout the body and affects a large proportion of our society, the neuroendocrine control mechanisms that accompany menopause are poorly understood. This review aims to examine rigorously the most accredited literature to provide an update about our current understanding of the role of the hypothalamic-pituitary axis in the onset of and transition into female reproductive senescence, focusing on the role of some specific neuropeptides in regulating the HPG axis and on their effects on several menopausal symptoms, especially referring to the cardiovascular risk, to open up new horizons for new therapeutic strategies. Modern medicine has significantly prolonged the lifespan of humans and most women spend one-third to half of their lifetime in post- menopause ,4. OvariThe reproductive neuroendocrine axis in postmenopausal women is intact and responds robustly (intensely) to the removal of ovarian hormones.In primates, menopause is the signal of the complete collapse of ovarian function .Decreased ovarian function is accompanied by the loss of circulating estradiol and progesterone, reduced steroid feedback and signaling to hypothalamic areas that modulate gonadotropin releasing hormone (GnRH) production and release and increased pituitary gonadotropin levels (FSH and LH) .The gonadotropin releasing hormone (GnRH) system is the primary hypothalamic regulator of reproduction and extensive studies have demonstrated the role of the GnRH system in age-related reproductive failure.This review wants to underline the role of kisspeptin, neurokinin B (NKB), calcitonin gene related peptide (CGRP) and several other neuropetides in female reproductive aging and some menopausal symptoms, mediating negative steroid feedback within the HPG axis and directly acting on different organs and tissues throughout the body.2-This review includes medical papers published in the English language since 1990 about the relationship between estrogens, some neuropeptides and HPG axis in pre, peri and postmenopausal women and their supposed role in menopausal symptoms, making particular reference to their effects on nasal mucosa and cardiovascular risk. All pertinent articles have been identified through a MEDLINE, PMC and EMBASE search and then reports have been selected through systematic review of all references.3-Despite the continued aging of the central nervous system, there is compelling evidence that many aspects of reproductive neuroendocrine function remain intact after menopause.After the menopause, there are age-related changes in the reproductive neuroendocrine axis: studies using indirect pharmacological methods provide evidence that GnRH secretion is increased in postmenopausal women compared to premenopausal women . ImportaThe hypothalamus has been shown to be the major site of steroid negative feedback in the human ,12.These studies demonstrate intact hypothalamic function in the postmenopausal period and indicate that removal of steroid negative feedback in postmenopausal women is linked to increased GnRH secretion from the hypothalamus. Moreover, GnRH mRNA is increased in the hypothalamus of postmenopausal women, as would be expected with removal of steroid negative feedback . The eleHybridization histochemistry studies have shown that the hypertrophied neurons express estrogen receptor α (ERα) and neurokinin B (NKB), kisspeptin (KiSS-1) and other neuropeptides’s mRNA and that menopause is associated with a striking increase in these neuropeptides’s gene expression , 17. RemIt has been proposed that these hypertrophied neurons participate in the hypothalamic circuitry regulating estrogen negative feedback, because the hypertrophy occurs in a subpopulation of Erα mRNA-expressing neurons in concert with estrogen withdrawal and gonadotropin hypersecretion .3.1-Studies pioneered by the endocrinologist Selmar Aschheim in 1964 showed that ovaries transplanted from young female rats with normal estrous cycles into old, non-cycling mice failed to restore estrous cyclicity, suggesting that the ovary is not the sole determinant of female reproductive senescence . In 1972More than four decades ago, Sheehan and Kovacs described pronounced differences in hypothalamic neuronal morphology between pre- and postmenopausal women . The neuin situ hybridization allowed characterization of mRNA expression in the hypertrophied neurons of postmenopausal women.The development of The hypertrophied neurons express ERα mRNA but do not express GnRH . The incHybridization of hypothalamic sections with a variety of cDNA probes revealed that the majority of hypertrophied neurons express, kisspeptin, neurokinin B (NKB), substance P (SP) and other tachykinins gene transcripts . Thus, tDynorphin mRNA is also expressed in the hypertrophied neurons, but unlike the elevation of NKB and kisspeptin gene expression in postmenopausal women, the expression of the mRNA encoding dynorphin is reduced .Similar to the human, menopause in monkeys is characterized by ovarian failure and gonadotropin hypersecretion , 30, 31.3.2-In 2007 , a key oDYN is an endogenous opioid peptide (EOP) that appears to mediate the inhibitory feedback control of progesterone on GnRH secretion , 36. In To simplify the reference, the name of this cell group has been abbreviated as the KNDy  subpopulation; each of the component neuropeptides  when examined in dual-label studies showed a very high degree of colocalization with gonadal hormone steroid receptors, specifically the α-isoform of the estrogen receptor (ERα) , 45, 46,Although each of these three peptides is found in separate sets of neurons in other brain regions, recent evidence suggests that the colocalization of KNDy peptides seen in the ARC is unique among brain regions and is conserved across multiple mammalian species that include the rat , mouse , sheep  and goatAlthough we have defined KNDy cells on the basis of the presence of three peptides, this subpopulation may well contain other neurotransmitters as well as other receptors.In the case of NKB/kisspeptin/dynorphin neurons, that neurotransmitter may be glutamate, based on the colocalization of vGLUT-2- immunoreactivity in the rat  and shee3.3-It’s necessary to show that KNDy cells directly contact GnRH neurons to understanding their functional role. Because colocalization of the three KNDy peptides in the ARC appears to be unique among brain areas examined to date, colocalization of multiple KNDy peptides in the same axon terminal can be used to determine the efferent targets of this subpopulation. Using this approach, it has been seen that NKB fibers project from the arcuate nucleus to both the internal and external zones of the median eminence, including the lateral palisade zone, a site with dense GnRH terminals: KNDy cells would provide direct inputs to GnRH neurons in the POA as well as in the mediobasal hypothalamus (MBH) at the level of their cell bodies and dendrites , 53.e.g. gonadal steroids). In either case, one would predict that appropriate postsynaptic receptors to KNDy peptides should also be expressed within KNDy cells. Actually, this appears to be true for at least two of the three KNDy peptide receptors: the high-affinity receptor for NKB (NK3R) is colocalized in KNDy cells in rats [Kiss1r) is expressed predominantly in GnRH cells, it is also found in other hypothalamic regions but, at least in mice, is not expressed in the ARC nucleus, suggesting a lack of colocalization within KNDy cells [One of the most interesting features of KNDy cells is that most appear to receive input from other KNDy cells and thus form a reciprocally interconnected network within the ARC , 47, 53: in rats , mice [3 in rats  and shee in rats  and the  in rats . AlthougDy cells .3.4 -The observation that KNDy neurons form an interconnected network presumably capable of producing a synchronous burst of firing has led to speculation that they may represent an important component of the multiunit activity known as the GnRH pulse generator , 43. It 2 and progesterone, respectively:because kisspeptin release from KNDy neurons is driving GnRH release during a pulse, E2 inhibition of kisspeptin expression would be expected to inhibit GnRH pulse amplitude. In contrast, DYN terminates and prevents GnRH secretion between pulses. Thus, a stimulation of DYN release by progesterone might be expected to prolong the interval between pulses and thus reduce GnRH pulse frequency.Each GnRH pulse is triggered by an initial increase in NKB from a few KNDy neurons, which stimulates further NKB release; the resulting positive feedback loop produces release of kisspeptin onto GnRH neurons and hence the extremely rapid increase in GnRH secretion that occurs at the onset of a pulse . NKB stiKISS1[KISS1[There is compelling evidence that changes in KNDy cells occur as a part of the process of normal aging in association with menopause. Specifically, it has long been known that in the brains of postmenopausal women, there is selective hypertrophy of neurons of the infundibular (arcuate) nucleus of the human hypothalamus . Rance aKISS1, NKB [16KISS1 as well KISS1mRNA; thuSS1[KISS1 in theseSS1[KISS1, consistSS1[KISS1. These cSS1[KISS1.4 –paradoxical nasal stuffiness”  . After this study, the effects of menopausal status on the content of vasoactive neuropeptides in the arterial wall were investigated .Cardiovascular disease (CVD), such as coronary artery disease (CAD) and hypertension (HTN), is more common in men than in premenopausal women (Pre-MW) of the same age, suggesting cardiovascular benefits of estrogen , 64. EpiThen, we analize recent studies about the effect of two neuropeptides (CGRP and orexin-A) on CVD in post-MW , 68 and,4.1 -Postmenopausal women often complain of reduced nasal function, both in terms of olfactive deficit and nasal blockage, even if no swelling or morphological alterations can be demonstrated . ElderlyThe activity of nasal mucosa in women is supposed to be related to variations in sex hormones,73. IndeNappi et al.  demonstrThis study has been the starting point to investigate the effects of menopausal status on the content of vasoactive neuropeptides in the arterial wall.4.2 -Autonomic pathways play a critical role in regulating vascular resistance by allowing selective redistribution of blood flow to those parts which have a greater demand . CathecoFrom January 2005–June 2005, Di Carlo et al. enrolled 20 premenopausal women  and 20 postmenopausal women , ≥12 months and ≤ 24 months after menopause, who were undergoing abdominal hysterectomy with bilateral oophorectomy for benign conditions to evaluate the presence of NPY, VIP, SP, E2, estrogen receptor α (ERα), and S100  in the uterine arteries of pre- and post- menopausal women of similar age . In Post4.3 –4.3a –Calcitonin gene-related peptide (CGRP), identified in 1982, belong to the calcitonin family of neuropeptides which also includes adrenomedullin (AM), amylin, intermedin, calcitonin and calcitonin receptor-stimulating peptide . CGRP isCGRP, estradiol (E(2)) and progesterone (P) are independent protective factors of CHD. There was no relationship between Hcy, CGRP and E(2), and P [In another study , it was ), and P .Current evidences of role of molecule CGRP in pathophysiology of menopausal symptoms really stimulate us further for the assessment of role CGRP receptor antagonists such as olcegepant and telcagepant in ameliorating various postmenopausal symptoms.4.3b –Orexins A and B  are recently discovered hypothalamic neuropeptides, involved in the regulation of feeding behaviour, sleep–wakefulness, and neuroendocrine homeostasis , 89. In El-Seedek et al. have reported postmenopausal women not on ERT had significantly higher plasma orexin-A levels, paralleling the significantly lower estrogen levels. Arterial blood pressure was also significantly higher in this group . It mighPostmenopausal women not receiving ERT had significantly higher plasma cholesterol and TG levels than reproductive-age women, but, more importantly, the levels were also higher than in those receiving ERT . OrexinsIt is also thought that orexins act in harmony with ghrelin, leptin and NPY, among others, to control energy balance and metabolism, and derangement in any of these can be associated with obesity .The conclusion is that plasma orexin-A levels are elevated after menopause. The association between this elevation and hypoestrogenism was suggested when normal values were found in matched postmenopausal women on ERT. A putative causal relationship is suggested between increased orexin levels and some of the manifestations of the menopausal syndrome, notably, an increased cardiovascular risk profile so, a possible inhibitory effect of estrogen on orexin might partially account for its cardioprotective effect .4.4 –Nowadays, whether menopausal hormone therapy (MHT) is beneficial in postmenopausal CVD remains controversial . DespiteBifulco et al. compared the effects of HRT and tibolone on nasal symptomatology/function and on local concentrations of estradiol and its receptor and neuropeptides  in postmenopausal women complaining of paradoxical nasal stuffiness. All treated patients reported an improvement in climacteric symptomatology; theevaluation of the immunopositivity for E2, estradiol receptor (ER), Substance P (SP), Vasoactive Intestinal Peptide (VIP) and neuropeptide Y (NPY) revealed HRT induced a significant increase in E2, ERα, VIP and SP and a decrease in NPY immunopositivity. Tibolone determined a significant increase in ERα, VIP and SP and a decrease in NPY immunopositivity without any action on E2 immunopositivity so, both treatments may modulate nasal mucosa function through an action on cholinergic, adrenergic and sensory peptides and improve nasal function and symptomatology in postmenopausal women with paradoxical nasal stuffiness.On the basis that the alterations seen in nasal mucosa of Post-MW with paradoxical nasal stuffiness may be a clinical model of the general effect of menopausal status on cardiovascular system, this study could suggest HRT could positively act on CVD risk, modulating the levels of vasoconstrictor (NPY) and vasodilator neuropeptides (VIP and SP).5 –This review demonstrates that a complex network between GnRH, FSH, LH, estrogens and several neuropeptides plays an important role in the transition into female reproductive aging and in the pathogenesis of some of the main menopausal symptoms .Understanding how the synthesis and release of individual KNDy peptide is orchestrated within KNDy neurons and their projections, and ultimately translated into control of the reproductive neuroendocrine axis, will likely have key relevance for a wide range of issues affecting reproductive health and disease.Estrogen and other estrogenic compounds have significant vasodilator and other beneficial vascular effects. Some of the effects of estrogen are mediated via vascular ER. As things stand now, medical evidence-based guidelines and the FDA recommendations are that MHT may be used for the relief of moderate to severe postmenopausal symptoms, but not for the routine prevention of chronic disease conditions and primary or secondary Coronary Heart Disease (CHD), with the exception that MHT may be used conditionally for the prevention of osteoporosis when other interventions have been considered and are deemed inappropriate.Nowadays, very little is known about the effects of estrogens on autonomic vasoconstrictor and vasodilator neuropeptides at the arterial level The report of a tight correlation between postmenopausal hypoestrogenism and the content in arterial-wall autonomic terminations of different vasoactive neuropeptides  open up new horizons for the comprehension of the biology of menopausal status and for the finding of new therapeutic strategies to treat several menopausal symptoms and reduce the menopausal CVD risk.


E. coli (EPEC and EHEC) continue to pose a threat to human health worldwide. While EPEC remains a significant cause of diarrhea in low-income countries, EHEC is more common as a food or water-borne pathogen in industrialized countries. A major difference between EPEC and EHEC is that EPEC has only a human reservoir of infection while EHEC is a zoonotic disease. Strains of EHEC are commensal in many ruminants, particularly cattle, and hence entry into the food chain through fecal contamination of food or water is a risk factor for infection. Another characteristic of EHEC but not EPEC is the production of Shiga toxins, which are associated with the development of severe complications of infection, namely hemorrhagic colitis (HC) and the hemolytic uremic syndrome (HUS). Whereas, HUS can affect patients of any age, EPEC remains a pathogen of infants less than 2 years of age  as EHEC such as O157:H7, although it role in defining EHEC is blurred somewhat by the existence of STEC that lack LEE yet cause HC and HUS and, as in the case of the recent EHEC O104:H4 outbreak, may be highly virulent (Frank et al., Despite their differing epidemiology and clinical manifestations, EPEC and EHEC are highly related and share many virulence determinants and features. Both pathogens share a distinctive mechanism of intestinal colonization known as attaching and effacing A/E lesion formation. A/E lesions are characterized by tight attachment of the bacteria to the enterocyte surface, the localized destruction of microvilli and massive ultrastructural changes underneath the adherent bacteria resulting from the accumulation of filamentous actin (Wong et al., The LEE T3SS system is similar to those found in other pathogens but has an additional filament to the T3SS needle made of polymerized EspA subunits. EspA is essential for translocation to occur and may also provide some adhesive function during bacterial colonization. The most widely accepted model is that the hydrophobic LEE-encoded proteins, EspB and EspD, form a pore in the host cell membrane that provide a conduit for effector translocation through the T3SS. Together EspA, EspB, and EspD constitute the LEE translocon. A recent proposal is that type III secretion is a two-step process whereby surface-localized effector proteins are part of an intermediate effector-translocator complex that precedes effector translocation (Pilar and Coombes, Among the many translocated effector proteins, Tir plays a critical role in intimate attachment through binding the outer membrane protein intimin. Although intimin and Tir alleles are largely interchangeable despite sequence variation among different A/E pathogens, intimin has been associated with additional Tir-independent adhesive functions. This has also been explored as a source of EPEC/EHEC tissue tropism in the past (Fitzhenry et al., While the intimin-Tir interaction has been intensely studied, not all effector proteins contribute to A/E lesion formation and cytoskeletal changes. It has recently emerged that several effector proteins play a role in dampening the inflammatory response. The effector proteins involved are all encoded outside the LEE and have novel enzymatic functions that target NF-κB and MAPK signaling. For example, NleE is a cysteine methyl transferase that methylates the zinc finger domain of TAB2/3, thereby inhibiting TAB2/3 interaction with ubiquitinated TRAF and blocking the phosphorylation of IκB by the IKK complex and hence the degradation of IκB (Newton et al., E. coli pathogens makes the study of other potential virulence factors critically important. For example, EhaJ is an autotransporter protein that is shared by strains of belonging to both EPEC and EHEC lineages. Autotransporters are highly prevalent in EPEC/EHEC genomes, and many contribute to inter-bacterial interactions and biofilm formation (Wells et al., E. coli, and this requires the function of a putative glycosyltransferase encoded by the adjacent gene, egtA. In addition, EhaJ has extracellular matrix binding properties, suggesting that the autotransporter may contribute to host infection. Other factors apart from the T3SS may also contribute to the ability of EHEC O157:H7 to persist and colonize the surface of plants, including pili and flagella (Saldana et al., Although much research is aimed at understanding the function of the LEE and the translocated effector proteins [recently reviewed in Wong et al. ], the caThe reservoir of EHEC in cattle deserves particular attention as many LEE-independent factors, such as autotransporters, may contribute to persistence in this natural host. Indeed recent work reported here on the interactions of EHEC O157:H7 with bovine rectal epithelial cells suggests that the bacteria are internalized though an intimin- and Tir-independent mechanism. The rectum is the main site of EHEC O157:H7 colonization in cattle and acts as an ongoing source of contamination through fecal shedding (Low et al., E. coli pathogens that all contribute to a common gene pool from which new pathogens, such as EHEC O104:H4, may emerge. Ongoing surveillance with high throughput genomics and molecular and infection studies are the keys to understanding the ecology and pathogenesis of EPEC and EHEC and are our only weapons against a fast evolving and dynamic group of pathogens.Because of its prominence in large outbreaks of disease, much research has focused on EHEC O157:H7 virulence. Nevertheless non-O157 EHEC, of which there are more than 100 serotypes, also have the capacity to cause HUS and we know far less about their genome structure and evolution (Coombes et al.,