Analysis of the gut microbiota in the old order amish and its relation to the metabolic syndrome

Obesity has been linked to the human gut microbiota; however, the contribution of gut bacterial species to the obese phenotype remains controversial because of conflicting results from studies in different populations. To explore the possible dysbiosis of gut microbiota in obesity and its metabolic complications, we studied men and women over a range of body mass indices from the Old Order Amish sect, a culturally homogeneous Caucasian population of Central European ancestry. We characterized the gut microbiota in 310 subjects by deep pyrosequencing of bar-coded PCR amplicons from the V1-V3 region of the 16S rRNA gene. Three communities of interacting bacteria were identified in the gut microbiota, analogous to previously identified gut enterotypes. Neither BMI nor any metabolic syndrome trait was associated with a particular gut community. Network analysis identified twenty-two bacterial species and four OTUs that were either positively or inversely correlated with metabolic syndrome traits, suggesting that certain members of the gut microbiota may play a role in these metabolic derangements.     

Proteomic analysis of the venom of Heterometrus longimanus (Asian black scorpion)

Venoms have evolved over millions of years into potent cocktails of bioactive peptides and proteins. These compounds can be of great value to the pharmaceutical industry for numerous clinical applications. In this study, a novel proteomic - bioinformatic approach was utilised, where chromatography followed by gel electrophoresis was utilised to separate the venom peptides/proteins of Heterometrus longimanus (Asian black scorpion). Purified peptides were analysed by tandem mass spectrometry, de novo sequenced and then homology matched against known peptides in the Swiss-Prot protein database. Numerous potentially biologically active peptide matches were discovered, and a simple scoring system applied to putatively assign functions to the peptides. As a validation of this approach, the functional composition of the experimentally derived proteome is similar to that of other scorpions, and contains a potent mix of toxins, antimicrobials and ionic channel inhibitors.     

Novel role of the muskelin-RanBP9 complex as a nucleocytoplasmic mediator of cell morphology regulation

The evolutionarily conserved kelch-repeat protein muskelin was identified as an intracellular mediator of cell spreading. We discovered that its morphological activity is controlled by association with RanBP9/RanBPM, a protein involved in transmembrane signaling and a conserved intracellular protein complex. By subcellular fractionation, endogenous muskelin is present in both the nucleus and the cytosol. Muskelin subcellular localization is coregulated by its C terminus, which provides a cytoplasmic restraint and also controls the interaction of muskelin with RanBP9, and its atypical lissencephaly-1 homology motif, which has a nuclear localization activity which is regulated by the status of the C terminus. Transient or stable short interfering RNA–based knockdown of muskelin resulted in protrusive cell morphologies with enlarged cell perimeters. Morphology was specifically restored by complementary DNAs encoding forms of muskelin with full activity of the C terminus for cytoplasmic localization and RanBP9 binding. Knockdown of RanBP9 resulted in equivalent morphological alterations. These novel findings identify a role for muskelin–RanBP9 complex in pathways that integrate cell morphology regulation and nucleocytoplasmic communication.     

A phylogenomic tree inferred with an inexpensive PCR-generated probe kit resolves higher-level relationships among Neptis butterflies (Nymphalidae: Limenitidinae)

Recent advances in obtaining reduced representation libraries for next-generation sequencing permit phylogenomic analysis of species-rich, recently diverged taxa. In this study, we performed sequence capture with homemade PCR-generated probes to study diversification among closely related species in a large insect genus to examine the utility of this method. We reconstructed the phylogeny of Neptis Fabricius, a large and poorly studied nymphalid butterfly genus distributed throughout the Old World. We inferred relationships among 108 Neptis samples using 89 loci totaling up to 84 519 bp per specimen. Our taxon sample focused on Palearctic, Oriental and Australasian species, but included 8 African species and outgroups from 5 related genera. Maximum likelihood and Bayesian analyses yielded identical trees with full support for almost all nodes. We confirmed that Neptis is not monophyletic because Lasippa heliodore (Fabricius) and Phaedyma amphion (Linnaeus) are nested within the genus, and we redefine species groups for Neptis found outside of Africa. The statistical support of our results demonstrates that the probe set we employed is useful for inferring phylogenetic relationships among Neptis species and likely has great value for intrageneric phylogenetic reconstruction of Lepidoptera. Based on our results, we revise the following two taxa: Neptis heliodore comb. rev. and Neptis amphion comb. rev.     

Aquaporins in complex tissues. I.Developmental patterns in respiratory and glandular tissues of rat

Developmentalexpression of aquaporin water transport proteins is not well understoodin respiratory tract or secretory glands; here we define aquaporinprotein ontogeny in rat. Expression of aquaporin-3 (AQP3), AQP4, andAQP5 proteins occurs within 2 wk after birth, whereas AQP1 firstappears before birth. In most tissues, aquaporin protein expressionincreases progressively, although transient high-level expression isnoted in distal lung (AQP4 at postnatal day+2) and trachea (AQP5 at postnatalday +21 and AQP3 at postnatal day+42). In mature animals, AQP5 is abundant in distallung and salivary glands, AQP3 and AQP4 are present in trachea, andAQP1 is present in all of these tissues except salivary glands.Surprisingly, all four aquaporin proteins are highly abundant innasopharynx. Unlike AQP1, corticosteroids did not induce expression ofAQP3, AQP4, or AQP5 in lung. Our results seemingly implicate aquaporinsin proximal airway humidification, glandular secretion, and perinatalclearance of fluid from distal airways. However, the studies underscorea need for detailed immunohistochemical characterizations anddefinitive functional studies.

     

Iridoid glucosides from Pedicularis

Pedicularis palustris contains the iridoid glucosides aucubin and gardoside methyl ester, and five hydroxy derivatives of 8-epi-deoxyloganin, together with boschnaloside and three hydroxy derivatives of boschnaloside. From P. silvatica, plantarenaloside, 8-epiloganin and euphroside have been isolated. Euphroside is also present in P. lapponica, in addition to aucubin and mussaenoside. Chemical evidence for the structure of euphroside is presented. Pedicularioside is shown to be identical with penstemoside, and the former name is thus redundant.     

Aquaporins in complex tissues: distribution of aquaporins 1-5 in human and rat eye

Multiple physiological fluid movements areinvolved in vision. Here we define the cellular and subcellular sitesof aquaporin (AQP) water transport proteins in human and rat eyes byimmunoblotting, high-resolution immunocytochemistry, and immunoelectronmicroscopy. AQP3 is abundant in bulbar conjunctival epithelium andglands but is only weakly present in corneal epithelium. In contrast, AQP5 is prominent in corneal epithelium and apical membranes of lacrimal acini. AQP1 is heavily expressed in scleral fibroblasts, corneal endothelium and keratocytes, and endothelium covering thetrabecular meshwork and Schlemm's canal. Although AQP1 is plentiful inciliary nonpigmented epithelium, it is not present in ciliary pigmentedepithelium. Posterior and anterior epithelium of the iris and anteriorlens epithelium also contain significant amounts of AQP1, but AQP0(major intrinsic protein of the lens) is expressed in lens fiber cells.Retinal Müller cells and astrocytes exhibit notableconcentrations of AQP4, whereas neurons and retinal pigment epitheliumdo not display aquaporin immunolabeling. These studies demonstrateselective expression of AQP1, AQP3, AQP4, and AQP5 in distinct ocularepithelia, predicting specific roles for each in the complex networkthrough which water movements occur in the eye.

     

Expression of polypeptide GalNAc-transferases in stratified epithelia and squamous cell carcinomas: immunohistological evaluation using monoclonal antibodies to three members of the GalNAc-transferase family

Mucin-type O-glycosylation is initiated by a large family of UDP- GalNAc: polypeptide N -acetyl-galactosaminyltransferases (GalNAc- transferases). Individual GalNAc-transferases appear to have different functions and Northern analysis indicates that they are differently expressed in different organs. This suggests that O-glycosylation may vary with the repertoire of GalNAc-transferases expressed in a given cell. In order to study the repertoire of GalNAc-transferases in situ in tissues and changes in tumors, we have generated a panel of monoclonal antibodies (MAbs) with well defined specificity for human GalNAc-T1, -T2, and -T3. Application of this panel of novel antibodies revealed that GalNAc- transferases are differentially expressed in different cell lines, in spermatozoa, and in oral mucosa and carcinomas. For example, GalNAc-T1 and -T2 but not -T3 were highly expressed in WI38 cells, and GalNAc-T3 but not GalNAc-T1 or -T2 was expressed in spermatozoa. The expression patterns in normal oral mucosa were found to vary with cell differentiation, and for GalNAc-T2 and -T3 this was reflected in oral squamous cell carcinomas. The expression pattern of GalNAc-T1 was on the other hand changed in tumors to either total loss or expression in cytological poorly differentiated tumor cells, where the normal undifferentiated cells lacked expression. These results demonstrate that the repertoire of GalNAc-transferases is different in different cell types and vary with cellular differentiation, and malignant transformation. The implication of this is not yet fully understood, but it suggests that specific changes in sites of O-glycosylation of proteins may occur as a result of changes in the repertoire of GalNAc-transferases.