Unknown Members Research Articles

Transcription factor RORα is critical for nuocyte development

Nuocytes are essential in innate type-2 immunity and contribute to the exacerbation of asthma responses. Here we show that nuocytes arise in the bone marrow and differentiate from common lymphoid progenitors, which makes them distinct new members of the lymphoid lineage. Nuocytes required interleukin 7 (IL-7), IL-33 and Notch signalling for development in vitro. Double negative 1 (DN1) and DN2 pro-T-cell progenitors maintained nuocyte potential in vitro, although the thymus was not essential for nuocyte development. Notably, the transcription factor Rorα was critical for nuocyte development and their role in parasitic worm expulsion.

Genetic Contributions to Intergroup Responses: A Cautionary Perspective

We live in an era where genomic informa-tion can be collected with the ease of a saliva sample and the cost of genotyping is plummeting (Hirschhorn and Daly, 2005; Quinque et al., 2006 ). Intergroup research-ers interested in incorporating biologi-cal approaches into their methodological toolbox are thus faced with the question of whether molecular genetics can provide novel insight into their understanding of people’s responses to members of other groups. In the current commentary, we stitch together human and animal neuro-science with insight from molecular biology to posit mechanisms through which genetic variation and life experience may give rise to responses during intergroup situations. We then discuss avenues for empirical inves-tigation and urge for responsible research practices that take into consideration the negative societal consequences that can result from overinterpreting genetic data.Intergroup phenomena, such as dis-crimination and ethnic violence, emerge at the interindividual level, and as a result, cannot be fully explained by intrapsychic processes. However, in order to understand the effects of intergroup influences on the individual, it is useful to examine the pro-cesses unfolding in the person’s mind and brain. For this reason, researchers have increasingly combined experimental social psychology and neuroimaging techniques to dissect the neural basis of affective and cognitive mechanisms that contribute to intergroup responses. This approach has revealed many interconnected, but disso-ciable, brain regions involved in perceiving, evaluating, and regulating behaviors toward other people, including the amygdala, fusi-form gyrus, anterior cingulate cortex, and various parts of the prefrontal cortex. For comprehensive reviews of the existing neu-roscience research pertaining to intergroup relations we direct readers to Amodio and Ratner (2011) , Cunningham and Van Bavel (2009), Ito and Bartholow (2009), and Kubota et al. (2012).To understand how these neuroimag-ing findings might relate to genetics, it is important to recognize that communica-tion between neurons is facilitated by neu-rochemicals, such as neurotransmitters (e.g., serotonin, dopamine), neurotrophic factors (e.g., BDNF), and hormones (e.g., cortisol, testosterone, and oxytocin). The enzymes that synthesize these molecules, the receptors to which they bind, and the reuptake mechanisms and enzymes that determine their availability are all pro-teins that are coded for by genes (Way and Gurbaxani, 2008). Thus, the cellular and molecular levels of analysis, although not currently considered by most intergroup researchers, have the potential to provide unique insight into how genetic variation might influence intergroup responses.Genes are biologically meaningful seg-ments of DNA ( Snyder and Gerstein, 2003 ). Each gene consists of a sequence of nucleo-tides. Frequent variations in the ordering, number, type, and repetition of nucleotides are called polymorphisms. A single gene can have many different types of polymor -phisms (den Dunnen and Antonarakis, 2001; Gibson and Muse, 2002). In order for polymorphisms to influence cellular functioning, their genetic code has to be transcribed into RNA and this RNA needs to be translated into amino acids. The type of amino acids that are produced and their configuration determine the form of the resultant proteins (e.g., enzymes, receptors; Crick, 1958).Gene expression occurs when bio-chemical processes within a cell stimulate transcription factors that bind to particu-lar DNA motifs (i.e., specified nucleotide sequences) in the promoter region of a gene. Extracellular events can control genomic responses through receptor-mediated chan-nels (Cole, 2009). Relevant to the present concerns, social stressors, such as interact-ing with unknown outgroup members, have been shown to elevate levels of the hormone cortisol (Page-Gould et al., 2008; Amodio, 2009). Other research indicates that cor-tisol binds to corticosteroid receptors and then the bound receptors translocate to the nucleus and act as transcription factors. One effect is that they bind to sites in the promoter of the serotonin transporter gene to trigger the synthesis of proteins that con-trol the reuptake of extracellular serotonin. The availability of serotonin has impor-tant effects on an individual’s emotional responses and also indirectly contributes to serotonin regulation by influencing hypo-thalamic–pituitary–adrenal (HPA) path-ways that release cortisol ( Glatz et al., 2003; Heisler et al., 2007; Way and Gurbaxani, 2008). Thus, at the level of gene expression there is a bidirectional relationship between the genetic code and the environment.The expression of a gene can also be influenced by epigenetic factors, such as DNA methylation and histone modification (Goldberg et al., 2007). DNA methylation occurs when a methyl group attaches to the promoter region and blocks DNA transcrip-tion. Environmental factors (e.g., stress) can increase DNA methylation, and thus have long-term effects on gene expression. Research in rodents has shown that DNA methylation can persist through the repro-ductive process, and as a result, can influence

Anti-Catholicism and Obscene Literature: The Case of Mrs. Mary Catharine Cadiere and its Context

As every historian knows, religious minorities and other ‘out-groups’ have repeatedly faced accusations of sexual misconduct and its consequences: seduction, the breaking of families, promiscuous fornication, participation in orgies, ‘unnatural vice’, incest, sadism and masochism. In the second or third century, Minucius Felix recorded such charges against the early Christians: they make ‘love almost before they are acquainted; everywhere they introduce a kind of religion of lust, a promiscuous “brotherhood” and “sisterhood” by which ordinary fornication, under cover of a hallowed name, is converted to incest’. The Cathars and other medieval heretics were accused of promiscuous, incestuous orgies. Across early modern Europe, witches at their sabbats, it was learnedly pronounced, copulated with the Devil himself and, indiscriminately, with unknown members of both sexes, even parents, brothers and sisters.

The MetJ regulon in gammaproteobacteria determined by comparative genomics methods

BackgroundWhole-genome sequencing of bacteria has proceeded at an exponential pace but annotation validation has lagged behind. For instance, the MetJ regulon, which controls methionine biosynthesis and transport, has been studied almost exclusively in E. coli and Salmonella, but homologs of MetJ exist in a variety of other species. These include some that are pathogenic (e.g. Yersinia) and some that are important for environmental remediation (e.g. Shewanella) but many of which have not been extensively characterized in the literature.ResultsWe have determined the likely composition of the MetJ regulon in all species which have MetJ homologs using bioinformatics techniques. We show that the core genes known from E. coli are consistently regulated in other species, and we identify previously unknown members of the regulon. These include the cobalamin transporter, btuB; all the genes involved in the methionine salvage pathway; as well as several enzymes and transporters of unknown specificity.ConclusionsThe MetJ regulon is present and functional in five orders of gammaproteobacteria: Enterobacteriales, Pasteurellales, Vibrionales, Aeromonadales and Alteromonadales. New regulatory activity for MetJ was identified in the genomic data and verified experimentally. This strategy should be applicable for the elucidation of regulatory pathways in other systems by using the extensive sequencing data currently being generated.

Communication of collective identity in secret social groups: Hypothesis of a holographic signature of member affiliation

This work describes the theoretical basis for a non‐phonemic method of detecting membership in secret social groups, such as cults, criminal gangs, drug cartels and terrorist cells. Beginning with the basic sociological proposition that every social group develops a distinctive socio‐cultural order, it is postulated that the primary features of group identity will be encoded, via a multi‐level socio‐psycho‐physiological process, into the field of bio‐emotional relations connecting group members. This information is communicated in all group‐related interactions by a holographic‐like process and recorded in the members’ brains as a unique signature of group affiliation. The group’s identity signature should be present in measurements of interaction between members as a latent commonality marking group affiliation. It is hypothesized that the identity signature is enfolded as a hologram in the energy spectra of vocal interactions among group members. In clandestine groups, the effort to remain hidden should enhance the signal strength of the identity signature. It is expected that detection of unknown members can be achieved using the spectral commonality identified in the analysis of recordings of vocal interactions among known members as a reference signature for comparison with those of the vocal spectra of suspected or potential members.

Monitoring associations between clade-level variation, overall community structure and ecosystem function in enhanced biological phosphorus removal (EBPR) systems using terminal-restriction fragment length polymorphism (T-RFLP)

The role of Candidatus “Accumulibacter phosphatis” (Accumulibacter) in enhanced biological phosphorus removal (EBPR) is well established but the relevance of different Accumulibacter clades to the performance of EBPR systems is unknown. We developed a terminal-restriction fragment length polymorphism (T-RFLP) technique to monitor changes in the relative abundance of key members of the bacterial community, including Accumulibacter clades, in four replicate mini-sequencing batch reactors (mSBRs) operated for EBPR over a 35-day period. The ability of the T-RFLP technique to detect trends was confirmed using fluorescence in situ hybridisation (FISH). EBPR performance varied between reactors and over time; by day 35, performance was maintained in mSBR2 whilst it had deteriorated in mSBR1. However, reproducible trends in structure–function relationships were detected in the mSBRs. EBPR performance was strongly associated with the relative abundance of total Accumulibacter. A shift in the ratio of the dominant Accumulibacter clades was also detected, with Type IA associated with good EBPR performance and Type IIC associated with poor EBPR performance. Changes in ecosystem function of the mSBRs in the early stages of the experiment were more closely associated with changes in the abundance of (unknown) members of the flanking community than of either Accumulibacter or Candidatus “Competibacter phosphatis”. This study therefore reveals a hitherto unrecorded and complex relationship between Accumulibacter clades, the flanking community and ecosystem function of laboratory-scale EBPR systems.

Detecting the Identity Signature of Secret Social Groups: Holographic Processes and the Communication of Member Affiliation

The principles of classical and quantum holography are used to develop the theoretical basis for a non-phonemic method of detecting membership in secret social groups, such as cults, criminal gangs, drug cartels, and terrorist cells. Grounded in the basic sociological premise that every group develops a distinctive sociocultural order, the theory postulates that the primary features of a group's collective identity will be encoded, via a multilevel socio-psycho-physiological process, into the field of bio-emotional relations connecting group members. The principles of classical holography (Gabor 1948) are used to describe how this information is recorded in the members' brains and communicated in interaction by a holographic-like process as a tacit signature of group affiliation—a constancy in group-related interactions. It is hypothesized that the identity signature is enfolded as a hologram in the energy spectra of vocal interactions among group members. The principles of quantum holography (Gabor 1946) are used to provide a description of the group's ongoing interactions as a dynamic process—a continuous, quantized series of snapshots (quantum holograms). Linked together, two forms of holography provide a model of how the group's collective identity evolves and is communicated over time to its members, as the group adapts to the ever-changing reality of its endogenous and exogenous circumstances. Using spectral analysis techniques, it should be possible to detect the group's identity signature in the recordings of relatively small samples of vocal interactions. In clandestine groups, the effort to remain hidden actually enhances the signal strength of the identity signature. It is predicted, therefore, that detection of unknown members can be achieved by using the spectral commonality identified in the analysis of recordings of vocal interactions among known members as a reference signature for comparison with those of the vocal spectra of suspected or potential members.

Overexpression of Foxf2 in adipose tissue is associated with lower levels of IRS1 and decreased glucose uptake in vivo

Many members of the forkhead genes family of transcription factors have been implicated as important regulators of metabolism, in particular, glucose homeostasis, e.g., Foxo1, Foxa3, and Foxc2. The purpose of this study was to exploit the possibility that yet unknown members of this gene family play a role in regulating glucose tolerance in adipocytes. We identified Foxf2 in a screen for adipose-expressed forkhead genes. In vivo overexpression of Foxf2 in an adipose tissue-restricted fashion demonstrated that such mice display a significantly induced insulin secretion in response to an intravenous glucose load compared with wild-type littermates. In response to increased Foxf2 expression, insulin receptor substrate 1 (IRS1) mRNA and protein levels are significantly downregulated in adipocytes; however, the ratio of serine vs. tyrosine phosphorylation of IRS1 seems to remain unaffected. Furthermore, adipocytes overexpressing Foxf2 have a significantly lower insulin-mediated glucose uptake compared with wild-type adipocytes. These findings argue that Foxf2 is a previously unrecognized regulator of cellular and systemic whole body glucose tolerance, at least in part, due to lower levels of IRS1. Foxf2 and its downstream target genes can provide new insights with regard to identification of novel therapeutic targets.

Accurate and Scalable Techniques for the Complex/Pathway Membership Problem in Protein Networks

A protein network shows physical interactions as well as functional associations. An important usage of such networks is to discover unknown members of partially known complexes and pathways. A number of methods exist for such analyses, and they can be divided into two main categories based on their treatment of highly connected proteins. In this paper, we show that methods that are not affected by the degree (number of linkages) of a protein give more accurate predictions for certain complexes and pathways. We propose a network flow-based technique to compute the association probability of a pair of proteins. We extend the proposed technique using hierarchical clustering in order to scale well with the size of proteome. We also show that top-k queries are not suitable for a large number of cases, and threshold queries are more meaningful in these cases. Network flow technique with clustering is able to optimize meaningful threshold queries and answer them with high efficiency compared to a similar method that uses Monte Carlo simulation.

Culture-Independent Characterization of Bacterial Communities Associated with the Cold-Water Coral Lophelia pertusa in the Northeastern Gulf of Mexico

Bacteria are recognized as an important part of the total biology of shallow-water corals. Studies of shallow-water corals suggest that associated bacteria may benefit the corals by cycling carbon, fixing nitrogen, chelating iron, and producing antibiotics that protect the coral from other microbes. Cold-water or deep-sea corals have a fundamentally different ecology due to their adaptation to cold, dark, high-pressure environments and as such have novel microbiota. The goal of this study was to characterize the microbial associates of Lophelia pertusa in the northeastern Gulf of Mexico. This is the first study to collect the coral samples in individual insulated containers and to preserve coral samples at depth in an effort to minimize thermal shock and evaluate the effects of environmental gradients on the microbial diversity of samples. Molecular analysis of bacterial diversity showed a marked difference between the two study sites, Viosca Knoll 906/862 (VK906/862) and Viosca Knoll 826 (VK826). The bacterial communities from VK826 were dominated by a variety of unknown mycoplasmal members of the Tenericutes and Bacteroidetes, whereas the libraries from VK906/862 were dominated by members of the Proteobacteria. In addition to novel sequences, the 16S rRNA gene clone libraries revealed many bacterial sequences in common between Gulf of Mexico Lophelia corals and Norwegian fjord Lophelia corals, as well as shallow-water corals. Two Lophelia-specific bacterial groups were identified: a cluster of gammaproteobacteria related to sulfide-oxidizing gill symbionts of seep clams and a group of Mycoplasma spp. The presence of these groups in both Gulf and Norwegian Lophelia corals indicates that in spite of the geographic heterogeneity observed in Lophelia-associated bacterial communities, there are Lophelia-specific microbes.

Comparison of Cyanide Degrading Enzymes Expressed from Genes of Fungal Origin

A variety of fungal species are known to degrade cyanide through the action of cyanide hydratase, a specialized nitrilases which hydrolyze cyanide to formamide. This work is a report on two unknown and un-characterized members from Neurospora crassa and Aspergillus nidulans. Recombinant forms of three cyanide hydratases (CHT) originated from N. crassa, Gibberella zeae, and A. nidulans were prepared after their genes were cloned with N-terminal hexahistidine purification tags, expressed in E. coli and purified using immobilized metal affinity chromatography. These enzymes were compared according to their pH activity profiles, and kinetic parameters. Although all three were similar, the N. crassa CHT has the widest pH range of activity above 50% and highest turnover rate (<TEX>$6.6{\times}10^8min^{-1}$</TEX>) among them. The CHT of A. nidulans has the highest Km value of the three nitrilases evaluated in here. Expression of CHT in both N. crassa and A. nidulans were induced by the presence of KCN, regardless of any presence of nitrogen sources. These data can be used to determine optimal procedures for the enzyme uses in the remediation of cyanide-containing wastes.

Aliskiren and Dual Therapy in Type 2 Diabetes Mellitus

The discovery 110 years ago that crude kidney extract elicited a pressor reaction provided the first evidence of the existence of the renin–angiotensin–aldosterone system, a major vasoactive system,1 and the complex actions and interactions of this important cascade are still unfolding. Over the past few decades, various components of the renin−angiotensin−aldosterone system have been elucidated more completely, and previously unknown members have been discovered.2 Drugs that specifically block various components of the renin−angiotensin−aldosterone system have been developed (Figure 1). Angiotensin-converting–enzyme (ACE) inhibitors block the conversion of angiotensin I to angiotensin II, and clinically available angiotensin-receptor blockers block the angiotensin II . . .

Diagnosing post-mortem treatments which inhibit DNA amplification from US MIAs buried at the Punchbowl

The US military is committed to recovering and identifying the remains of unknown military service members. Casualties of the Korean War were exhumed from the National Memorial Cemetery of the Pacific, or Punchbowl, and submitted to the Armed Forces DNA Identification Laboratory (AFDIL) for mtDNA sequencing. Contrary to AFDIL's experience on other samples from this era, most failed to yield amplifiable DNA. Suspicion fell on mortuary practices that may have been applied to the remains, evidenced by a white powder found with the bones, and general records suggesting the use of formaldehyde-based stablizing agents. To improve the chances of successful identification of the unknown individuals, we looked for the causes underlying this failure. We did this by examining the state of the collagen, the most abundant biomolecule in bone, by using differential scanning calorimetry (DSC) and transmission electron microscopy (TEM). The DSC analyses showed collagens with a range of different thermal stabilities. When these results were compared with the DNA amplification results, a clear correlation between elevated thermal stability and amplification failure was evident. TEM analysis revealed that fibril integrity was maintained after thermal and acid treatments in the samples which failed amplification. Together these two approaches implicate a stabilization agent as the cause of problems with DNA analysis, presumably due to excessive cross-linking. Following the initial study, the ability of DSC to rapidly identify problem samples was tested in a blind study of 14 samples, the method successfully identifying all the problematic samples from Punchbowl. Within this unusual context, DSC analysis is a useful method to assess the likelihood of successful DNA extraction and amplification.

Novel synthetic route to liquid crystalline 4,4′‐bis(n‐alkoxy)azoxybenzenes: spectral characterisation, mesogenic behaviour and crystal structure of two new members

A simple synthetic method has been developed for the preparation of long‐chain 4,4′‐bis(n‐alkoxy)azoxybenzenes by reductive coupling of 4‐n‐alkoxynitrobenzenes using zinc powder and ammonium chloride in aqueous ethanol medium at ambient temperature. The new method was employed to synthesise known members (n = 1–12) of the 4,4′‐bis(n‐alkoxy)azoxybenzene (C n H2n+1OPhN(O) = NPhOC n H2n+1) series and also two hitherto unknown members (n = 13–14) of the series. The new compounds were characterised using spectral (IR, UV–visible, 1H NMR and FAB‐MS) data. The mesogenic behaviour of both compounds was studied by polarising optical microscopy, differential scanning calorimetry and small‐angle X‐ray diffraction techniques. The crystal structure of 4,4′‐bis(n‐tetradecyloxy)azoxybenzene was determined using single‐crystal X‐ray diffraction data. The packing of the molecules in the crystalline state is found to be a precursor to the smectic C phase structure.

Accumulation of apocarotenoids in mycorrhizal roots of leek ( Allium porrum)

Colonization of the roots of leek ( Allium porrum L.) by the arbuscular mycorrhizal fungus Glomus intraradices induced the formation of apocarotenoids, whose accumulation has been studied over a period of 25 weeks. Whereas the increase in the levels of the dominating cyclohexenone derivatives resembles the enhancement of root length colonization, the content of mycorradicin derivatives remains relatively low throughout. Structural analysis of the cyclohexenone derivatives by mass spectrometry and NMR spectroscopy showed that they are mono- and diglycosides of 13-hydroxyblumenol C and blumenol C acylated with 3-hydroxy-3-methyl-glutaric and/or malonic acid. Along with the isolation of three known compounds five others are shown to be hitherto unknown members of the fast-growing family of mycorrhiza-induced cyclohexenone conjugates.

Identifying Genes of Gene Regulatory Networks Using Formal Concept Analysis

In order to understand the behavior of a gene regulatory network, it is essential to know the genes that belong to it. Identifying the correct members (e.g., in order to build a model) is a difficult task even for small subnetworks. Usually only few members of a network are known and one needs to guess the missing members based on experience or informed speculation. It is beneficial if one can additionally rely on experimental data to support this guess. In this work we present a new method based on formal concept analysis to detect unknown members of a gene regulatory network from gene expression time series data. We show that formal concept analysis is able to find a list of candidate genes for inclusion into a partially known basic network. This list can then be reduced by a statistical analysis so that the resulting genes interact strongly with the basic network and therefore should be included when modeling the network. The method has been applied to the DNA repair system of Mycobacterium tuberculosis. In this application, our method produces comparable results to an already existing method of component selection while it is applicable to a broader range of problems.

Cover Picture: Electrophoresis 4/2008

AbstractThis special issue on capillary electrochromatography (CEC) and electrokinetic capillary chromatography (EKC) provides the reader with the latest developments and improvements in these two closely related micro‐column separation techniques.Issue 4 also offers one Fast Track article describing particularly important investigations in electrophoresis:Identification of unknown protein complex members by radiolocalization and analysis of low‐abundance complexes resolved using native polyacrylamide gel electrophoresisMahuya Bose, Brian P. Adams, Randy M. Whittal, Himangshu S. Bose

Identification of unknown protein complex members by radiolocalization and analysis of low‐abundance complexes resolved using native polyacrylamide gel electrophoresis

Identification of unknown binding partners of a protein of interest can be a difficult process. Current strategies to determine protein binding partners result in a high amount of false-positives, requiring use of several different methods to confirm the accuracy of the apparent association. We have developed and utilized a method that is reliable and easily substantiated. Complexes are isolated from cell extract after exposure to the radiolabeled protein of interest, followed by resolution on a native polyacrylamide gel. Native conformations are preserved, allowing the complex members to maintain associations. By radiolabeling the protein of interest, the complex can be easily identified at detection levels below the threshold of Serva Blue, Coomassie, and silver stains. The visualized radioactive band is analyzed by MS to identify binding partners, which can be subsequently verified by antibody shift and immunoprecipitation of the complex. By using this method we have successfully identified binding partners of two proteins that reside in different locations of a cellular organelle.

Understanding signaling in yeast: Insights from network analysis

Reconstruction of protein interaction networks that represent groups of proteins contributing to the same cellular function is a key step towards quantitative studies of signal transduction pathways. Here we present a novel approach to reconstruct a highly correlated protein interaction network and to identify previously unknown components of a signaling pathway through integration of protein-protein interaction data, gene expression data, and Gene Ontology annotations. A novel algorithm is designed to reconstruct a highly correlated protein interaction network which is composed of the candidate proteins for signal transduction mechanisms in yeast Saccharomyces cerevisiae. The high efficiency of the reconstruction process is proved by a Receiver Operating Characteristic curve analysis. Identification and scoring of the possible linear pathways enables reconstruction of specific sub-networks for glucose-induction signaling and high osmolarity MAPK signaling in S. cerevisiae. All of the known components of these pathways are identified together with several new "candidate" proteins, indicating the successful reconstructions of two model pathways involved in S. cerevisiae. The integrated approach is hence shown useful for (i) prediction of new signaling pathways, (ii) identification of unknown members of documented pathways, and (iii) identification of network modules consisting of a group of related components that often incorporate the same functional mechanism.

Abundance and functional diversity of riboswitches in microbial communities.

BackgroundSeveral recently completed large-scale enviromental sequencing projects produced a large amount of genetic information about microbial communities ('metagenomes') which is not biased towards cultured organisms. It is a good source for estimation of the abundance of genes and regulatory structures in both known and unknown members of microbial communities. In this study we consider the distribution of RNA regulatory structures, riboswitches, in the Sargasso Sea, Minnesota Soil and Whale Falls metagenomes.ResultsOver three hundred riboswitches were found in about 2 Gbp metagenome DNA sequences. The abundabce of riboswitches in metagenomes was highest for the TPP, B12 and GCVT riboswitches; the S-box, RFN, YKKC/YXKD, YYBP/YKOY regulatory elements showed lower but significant abundance, while the LYS, G-box, GLMS and YKOK riboswitches were rare. Regions downstream of identified riboswitches were scanned for open reading frames. Comparative analysis of identified ORFs revealed new riboswitch-regulated functions for several classes of riboswitches. In particular, we have observed phosphoserine aminotransferase serC (COG1932) and malate synthase glcB (COG2225) to be regulated by the glycine (GCVT) riboswitch; fatty acid desaturase ole1 (COG1398), by the cobalamin (B12) riboswitch; 5-methylthioribose-1-phosphate isomerase ykrS (COG0182), by the SAM-riboswitch. We also identified conserved riboswitches upstream of genes of unknown function: thiamine (TPP), cobalamine (B12), and glycine (GCVT, upstream of genes from COG4198).ConclusionThis study demonstrates applicability of bioinformatics to the analysis of RNA regulatory structures in metagenomes.