Nanodisc-associated acetylcholinesterase as a novel model system of physiological relevant membrane-bound cholinesterases. Inhibition by phenolic compounds.

Acetylcholinesterase (AChE) plays a pivotal role in the cholinergic system, and its inhibition is sought after in a wide range of applications, from insect control to Alzheimer's disease treatment. While the primary physiological isoforms of AChE are membrane-bound proteins, most assays for discovering new, safer, and potent inhibitors are conducted using commercially available soluble isoforms, such as the electric eel AChE (eeAChE). In this study, we conducted a comparative analysis of the activity and selectivity to phenolic inhibitors of recombinant human AChE, eeAChE and a mutant variant of human AChE known as dAChE4.

Mobile barrier mechanisms for Na-coupled symport in an MFS sugar transporter.

While many 3D structures of cation-coupled transporters have been determined, the mechanistic details governing the obligatory coupling and functional regulations still remain elusive. The bacterial melibiose transporter (MelB) is a prototype of major facilitator superfamily transporters. With a conformation-selective nanobody, we determined a low-sugar affinity inward-facing Na-bound cryoEM structure.

Mobile barrier mechanisms for Na-coupled symport in an MFS sugar transporter.

While many 3D structures of cation-coupled transporters have been determined, the mechanistic details governing the obligatory coupling and functional regulations still remain elusive. The bacterial melibiose transporter (MelB) is a prototype of the Na-coupled major facilitator superfamily transporters. With a conformational nanobody (Nb), we determined a low-sugar affinity inward-facing Na-bound cryoEM structure.

Novel cricothyrotomy assessment tool for attending physicians: A multicenter study of an error avoidance checklist.

Background: This study used existing literature and expert feedback to develop and pilot a novel error-avoidance checklist tool for cricothyrotomy in attending physicians. Prior literature has not focused on expert cricothyrotomy performance. While published checklists teach a specific procedural method, ideal for novice learners, this may hinder expert learners.

Human cystic fibrosis monocyte derived macrophages display no defect in acidification of phagolysosomes when measured by optical nanosensors.

Background: Defective macrophage phagolysosomal acidification is implicated in numerous lung diseases including Cystic Fibrosis (CF) and may contribute to defective pathogen killing. Conflicting reports relating to phagolysosomal pH in CF macrophages have been published, in part related to the use of pH-sensitive fluorescent probes where potential inadequacies in experimental design can be a contributing factor (e.g.

Confocal Raman spectroscopy of whole hairs.

This paper describes the application of Raman spectroscopy to whole hair fibers. Previously this has proved difficult because the hairs are relatively opaque, and spatial resolution diminishes with depth because of the change in refractive index. A solution is to couple confocal Raman with multivariate curve resolution (MCR) data analysis, which separates spectral differences with depth despite this reduction in resolution.

BARREN STALK FASTIGIATE1 is an AT-hook protein required for the formation of maize ears.

Ears are the seed-bearing inflorescences of maize (Zea mays) plants and represent a crucial component of maize yield. The first step in the formation of ears is the initiation of axillary meristems in the axils of developing leaves. In the classic maize mutant barren stalk fastigiate1 (baf1), first discovered in the 1950s, ears either do not form or, if they do, are partially fused to the main stalk.

The control of axillary meristem fate in the maize ramosa pathway.

Plant axillary meristems are composed of highly organized, self-renewing stem cells that produce indeterminate branches or terminate in differentiated structures, such as the flowers. These opposite fates, dictated by both genetic and environmental factors, determine interspecific differences in the architecture of plants. The Cys(2)-His(2) zinc-finger transcription factor RAMOSA1 (RA1) regulates the fate of most axillary meristems during the early development of maize inflorescences, the tassel and the ear, and has been implicated in the evolution of grass architecture.

Delayed presentation of traumatic diaphragmatic rupture.

We present the case of a 57-year-old woman who suffered a delayed presentation of a traumatic diaphragmatic rupture (TDR). Because TDR is frequently missed on initial presentation, a delay between trauma and diagnosis can present a unique diagnostic dilemma for the emergency physician. Important historical clues and physical examination findings are the keys to making this diagnosis.

Comparative proteomic analysis to study molecular events during gonad development in mice.

Sex determination represents a critical bifurcation in the road of embryonic development. It is based on a finely regulated network of gene activity, as well as protein-protein interactions and activation or silencing of signaling pathways. Despite the identification of a number of critical genes, many aspects of the molecular cascade that drives the differentiation of the embryonic gonad into either a testis or an ovary remain poorly understood.

Neuroleptic malignant syndrome associated with olanzapine therapy: a case report.

We present the case of a 42-year-old male with a history of schizophrenia who developed signs and symptoms consistent with Neuroleptic Malignant Syndrome (NMS) after 3 weeks of treatment with Olanzapine. The patient presented with hyperpyrexia, tremors, labile blood pressure, and mental status changes that had progressed over the preceding 24 h. Laboratory data revealed a metabolic acidosis and an escalating creatinine phosphokinase.

Suppression of the ndv mutant phenotype of Rhizobium meliloti by cloned exo genes.

The ndvA and ndvB genes of Rhizobium meliloti are involved in the export and synthesis, respectively, of the small cyclic polysaccharide beta(1,2)glucan. We have previously shown that spontaneous symbiotic pseudorevertants of ndv mutants do not produce periplasmic beta(1,2)glucan. Here we show that the pseudorevertants also do not produce extracellular beta(1,2)glucan, but do show alterations in the amount of the major acidic exopolysaccharide produced.

The ndvB locus of Rhizobium meliloti encodes a 319-kDa protein involved in the production of beta-(1----2)-glucan.

The ndvB locus of Rhizobium meliloti was sequenced and found to encode a 319-kDa protein involved in the production of beta-(1----2)-glucan. Transposon Tn5 mutagenesis revealed that a large portion of the downstream half of this gene is not essential for symbiosis but is required for optimal production of beta-(1----2)-glucan. A high molecular weight inner membrane protein, believed to be the ndvB gene product, was absent from two different upstream ndvB::Tn5 mutants.

Construction of a gene bank of Rhizobium leguminosarum Rld 164.

The total genomic DNA of R. leguminosarum Rld164 (trp, sms, azi) was cloned in the EcoR1 site of the wide host and conjugally transferable cosmid vector pLAFR1. The average insert size in the gene clones of the bank was found to be 21.

The ndvA gene product of Rhizobium meliloti is required for beta-(1----2)glucan production and has homology to the ATP-binding export protein HlyB.

The ndvA locus of Rhizobium meliloti is homologous to and can substitute for the chvA locus of Agrobacterium tumefaciens. We have previously shown that an ndvA mutant exhibited reduced motility and formed small, white, empty nodules on alfalfa roots. Here we show that this ndvA mutant is defective in the production of the cyclic extracellular polysaccharide beta-(1----2)glucan, even though a 235,000-dalton protein intermediate, known to be involved in the synthesis of this molecule, is present and active in vitro.

Common regulatory elements control symbiotic and microaerobic induction of nifA in Rhizobium meliloti.

We have previously demonstrated that the nifA promoter (nifAp) of Rhizobium meliloti is inducible under microaerobic conditions in the absence of alfalfa. Here we show that microaerobic activation of nifAp involves both cis- and trans-acting regulatory controls identical to those used symbiotically. The start site for nifA mRNA synthesis was found to be the same during symbiosis and microaerobiosis, and a deletion analysis of nifAp demonstrated that DNA between positions -62 and -45 is essential for induction.

Rhizobium meliloti genes required for nodule development are related to chromosomal virulence genes in Agrobacterium tumefaciens.

Symbiotically essential genes have been identified in Rhizobium meliloti that are structurally and functionally related to chromosomal virulence (chv) genes of Agrobacterium tumefaciens. Homologous sequences also exist in the genomes of other fast-growing rhizobia including Rhizobium trifolii, Rhizobium leguminosarum, and Rhizobium phaseoli. In Agrobacterium, the chvA and chvB loci are known to be essential for oncogenic transformation of dicotyledonous plants and for attachment to plant cells [Douglas, C.

Multiple mechanisms generate extrachromosomal circular DNA in Chinese hamster ovary cells.

Seven cloned small circular DNA molecules from CHO cells were sequenced and examined for the presence of homologies to each other and to a number of other functional sequences present in transposable elements, retroviruses, mammalian repeat sequences, and introns. The sequences of the CHO cell circular DNA molecules did not reveal common structural features that could explain their presence in the circular DNA population. A gene bank was constructed for CHO chromosomal DNA and sequences homologous to two of the seven small circular DNA molecules were isolated and sequenced.

Cloning and characterization of small circular DNA from Chinese hamster ovary cells.

Small polydisperse circular (spc) DNA was isolated and cloned, using BglII from Chinese hamster ovary (CHO) cells. The properties of 47 clones containing at least 43 different BglII fragments are reported. The majority of the clones probably contain entire sequences from individual spcDNA molecules.

Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti.

A broad host range cloning vehicle that can be mobilized at high frequency into Gram-negative bacteria has been constructed from the naturally occurring antibiotic resistance plasmid RK2. The vehicle is 20 kilobase pairs in size, encodes tetracycline resistance, and contains two single restriction enzyme sites suitable for cloning. Mobilization is effected by a helper plasmid consisting of the RK2 transfer genes linked to a ColE1 replicon.

Small circular deoxyribonucleic acid of Drosophila melanogaster: homologous transcripts in the nucleus and cytoplasm.

We have recently characterized small circular DNA of Drosophila cultured cells in terms of its average size, sequence complexity, and homology to intermediate repetitive DNA. We show here that transcripts homologous to small circular DNA are present in various RNA fractions. Nuclear poly(A+), nuclear poly(A-), and polysomal poly(A+) RNA drive 10, 7, and 20%, respectively, of in vitro labeled small circular DNA tracer into hybrid.

Small circular DNA of Drosophila melanogaster: chromosomal homology and kinetic complexity.

Nucleic acid reassociation techniques were used to determine the kinetic complexity of small circular DNA in cultured cells of Drosophila melanogaster. Two kinetic components are present. One of these constitutes 82% of the mass of the circular DNA and has a complexity of 1.