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Meet Smudge. Our Founder.
At Catipilla, we believe that all cats should be able to climb to fulfil an innate instinct. Catipilla promotes a healthy lifestyle by replicating a cat's natural environment and creating safe spaces for them to climb and roam freely.
If you want to be one of the first people in the world to own a Catipilla, you can now order and reserve a unit from our first production run. These are being offered at a reduced rate and on a first come, first served basis.
Our Product
Catipilla is a new alternative to the traditional climbing frame. It's modular design is unique and can be configured to suit the needs of any cat. Our modules can be fixed onto any wall, making great use of vertical space.


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Kinabatangan Orang utan Conservation Project
Kinabatangan Orang utan Conservation Project
Meijaard E.,People and Nature Consulting International |
Meijaard E.,Australian National University |
Wich S.,Sumatran Orangutan Conservation Program PanEco YEL |
Wich S.,University of Zürich |
And 2 more authors.
Annals of the New York Academy of Sciences | Year: 2012
Orangutan survival is threatened by habitat loss and illegal killing. Most wild populations will disappear over the next few decades unless threats are abated. Saving orangutans is ultimately in the hands of the governments and people of Indonesia and Malaysia, which need to ensure that habitats of viable orangutan populations are protected from deforestation and well managed to ensure no hunting takes place. Companies working in orangutan habitat also have to play a much bigger role in habitat management. Although the major problems and the direct actions required to solve them-reducing forest loss and hunting-have been known for decades, orangutan populations continue to decline. Orangutan populations in Sumatra and Borneo have declined by between 2,280 and 5,250 orangutans annually over the past 25 years. As the total current population for the two species is some 60,000 animals in an area of about 90,000 km 2, there is not much time left to make conservation efforts truly effective. Our review discusses what has and has not worked in conservation to guide future conservation efforts. © 2012 New York Academy of Sciences.
Meijaard E.,People and Nature Consulting International |
Meijaard E.,Australian National University |
Mengersen K.,Queensland University of Technology |
Buchori D.,The Nature Conservancy Indonesia Program |
And 13 more authors.
PLoS ONE | Year: 2011
Species conservation is difficult. Threats to species are typically high and immediate. Effective solutions for counteracting these threats, however, require synthesis of high quality evidence, appropriately targeted activities, typically costly implementation, and rapid re-evaluation and adaptation. Conservation management can be ineffective if there is insufficient understanding of the complex ecological, political, socio-cultural, and economic factors that underlie conservation threats. When information about these factors is incomplete, conservation managers may be unaware of the most urgent threats or unable to envision all consequences of potential management strategies. Conservation research aims to address the gap between what is known and what knowledge is needed for effective conservation. Such research, however, generally addresses a subset of the factors that underlie conservation threats, producing a limited, simplistic, and often biased view of complex, real world situations. A combination of approaches is required to provide the complete picture necessary to engage in effective conservation. Orangutan conservation (Pongo spp.) offers an example: standard conservation assessments employ survey methods that focus on ecological variables, but do not usually address the socio-cultural factors that underlie threats. Here, we evaluate a complementary survey method based on interviews of nearly 7,000 people in 687 villages in Kalimantan, Indonesia. We address areas of potential methodological weakness in such surveys, including sampling and questionnaire design, respondent biases, statistical analyses, and sensitivity of resultant inferences. We show that interview-based surveys can provide cost-effective and statistically robust methods to better understand poorly known populations of species that are relatively easily identified by local people. Such surveys provide reasonably reliable estimates of relative presence and relative encounter rates of such species, as well as quantifying the main factors that threaten them. We recommend more extensive use of carefully designed and implemented interview surveys, in conjunction with more traditional field methods. © 2011 Meijaard et al.
Bruford M.W.,University of Cardiff |
Ancrenaz M.,Kinabatangan Orang utan Conservation Project |
Chikhi L.,CNRS Biological Evolution and Diversity Laboratory |
Lackman-Ancrenaz I.,Kinabatangan Orang utan Conservation Project |
Goossens B.,University of Cardiff
Endangered Species Research | Year: 2010
Genetic management of fragmented populations poses logistical and theoretical challenges to conservation managers. Simulating changes in genetic diversity and differentiation within and among fragmented population units under different management scenarios has until now rarely used molecular marker data collected from present-day populations. Here we examine the genetic implications of management options for the highly fragmented yet globally significant orang-utan population in the Lower Kinabatangan Wildlife Sanctuary, Sabah, Malaysia. We simulated the effects of non-intervention, translocation, corridor establishment and a mixture of the latter 2 approaches on future genetic diversity in this population using the stochastic simulation software VORTEX and a well-described molecular dataset for 200 individuals from within the Sanctuary. We found that nonintervention resulted in high extinction risks for a number of subpopulations over short demographic timescales (<5 generations). Furthermore, the exclusive use of either translocation or corridor establishment as a management tool was insufficient to prevent substantial levels of inbreeding using demographically and logistically feasible translocation rates and was insufficient to prevent inbreeding and extinction in the most isolated subpopulations using conservative corridor establishment rates. Instead, a combination of modest translocation rates (1 ind. every 20 yr) and corridor establishment enabled even the most isolated subpopulations to retain demographic stability and constrain localised inbreeding to levels below a threshold of 0.1. Our simulations suggest that this mixed management approach is both a pragmatic and potentially successful course of action and that this combination may be useful in other species and fragmented populations in the future. The use of present-day molecular data in stochastic simulations requires further development, but here we show that it can aid predictive modelling. © Inter-Research 2010.
Matsuda I.,Kyoto University |
Ancrenaz M.,Kinabatangan Orang utan Conservation Project |
Akiyama Y.,Japan National Institute for Land and Infrastructure Management |
Majalap N.,Sabah Forest Research Center |
Bernard H.,Universiti Malaysia Sabah
Ecological Research | Year: 2014
Although the importance of natural licks for terrestrial mammals is widely acknowledged, we report here for the first time its importance for large terrestrial mammals in a degraded riverine forest in Borneo. Our results clearly demonstrated that various mammals, including bearded pig, sambar deer, and endangered orang-utans, were using the natural lick, though large arboreal/avian herbivore/omnivore animals were not attracted to the natural lick. In addition, the diversity of mammal species recorded in this study was lower than those recorded in the dry lowland forest. Possible reasons for this difference between the different forest types are discussed. © 2014, The Ecological Society of Japan.
Meijaard E.,People and Nature Consulting International |
Meijaard E.,University of Queensland |
Meijaard E.,Australian National University |
Buchori D.,The Nature Conservancy Indonesia Forest Program |
And 33 more authors.
PLoS ONE | Year: 2011
Human-orangutan conflict and hunting are thought to pose a serious threat to orangutan existence in Kalimantan, the Indonesian part of Borneo. No data existed prior to the present study to substantiate these threats. We investigated the rates, spatial distribution and causes of conflict and hunting through an interview-based survey in the orangutan's range in Kalimantan, Indonesia. Between April 2008 and September 2009, we interviewed 6983 respondents in 687 villages to obtain socio-economic information, assess knowledge of local wildlife in general and orangutan encounters specifically, and to query respondents about their knowledge on orangutan conflicts and killing, and relevant laws. This survey revealed estimated killing rates of between 750 and 1800 animals killed in the last year, and between 1950 and 3100 animals killed per year on average within the lifetime of the survey respondents. These killing rates are higher than previously thought and are high enough to pose a serious threat to the continued existence of orangutans in Kalimantan. Importantly, the study contributes to our understanding of the spatial variation in threats, and the underlying causes of those threats, which can be used to facilitate the development of targeted conservation management. © 2011 Meijaard et al.

subset: commoncrawl


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David Bowie's current Reality tour has provided the perfect platform for BSS Audio to launch its new SB2 software, which has been successfully controlling ten FDS-366T Omnidrive Compact Plus devices over a wireless LAN.
Liverpool-based Adlib Audio, which recently provided stacks, racks and control to Bowie for the inaugural three-month UK/European leg of his world tour, was quick to upgrade its existing stock of 366s to incorporate the new WHISEWORKS-NTM(tm) filtering method. The deci
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Not much new to show in #100DragonDays, which is good because I broke my poor little PowerShot. I like my phone a lot, but its camera is really terrible.
I think I've put about as much filler in as I'm going to.
Time to get messy!
(Prediction: There will be a bit of stalling in #The100DayProject before I actually mix up a batch of paper mache and commit.)


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SET 1: Suzy Greenberg[1] > Buried Alive, Punch You In the Eye, Uncle Pen, Horn > Chalk Dust Torture, Esther[2] > Dinner and a Movie > Bouncing Around the Room > Run Like an Antelope[3]
SET 2: Axilla, The Curtain > Stash -> Manteca -> Stash -> The Lizards, Bathtub Gin -> Hold Your Head Up > Cracklin' Rosie > Hold Your Head Up, The Squirming Coil, Big Black Furry Creature from Mars
ENCORE: Sweet Adeline, Good Times Bad Times[4] -> Paul and Silas[5] -> Pig in a Pen[6]
Add a Review
Trey Anastasio Band: September 17, 2017
5 days ago
Waterfront Park
Set 1: Mozambique, Cayman Review, Curlew's Call, Sand, Magilla, Everything's Right, Valentine, Money, Love and Change, Shine, Burlap Sack and Pumps, Dark and Down, Clint Eastwood, First Tube[1]
Encore: Water in the Sky[2], Push On 'Til the Day[3]
[1] Page on keys.
[2] Trey, Mike on bass, and Grace Potter on vocals.
[3] Dave Grippo on saxophone.
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This property has been removed by the agent. It may be sold or temporarily removed from the market
5 bedroom detached house for saleBroad Lane, Hale
Guide Price £1,500,000
Property Description
Full description
A stunning, unique property set in 0.36 acres of mature landscaped gardens on one of the most sought after roads in Hale. The accommodation comprises, Covered porch, entrance hall, cloakroom and wc, living room, dining area, sitting room, dining/kitchen, utility room, master bedroom suite, four further bedrooms (one en suite), family bathroom. Double garage. EPC -E. Council Tax Band G.
Entrance Hall - A welcoming entrance hall with large windows to the front elevation, bare brick walls and tiled flooring. Access to a cloakroom and wc and the rest of the ground floor. A curved staircase leads to the first floor.
Drawing Room And Dining Room - This room provides the perfect reception space for entertaining. With large floor to ceiling windows and sliding patio doors creating a bright and spacious room. Tiled flooring throughout and a spiral staircase leading to the master suite.
Sitting Room - An ideal second sitting room with large windows to the side and rear elevations and a door leading to the rear patio area.
Breakfast Kitchen - A stunning breakfast kitchen, well appointed with a range of base and eye level units with corian work tops. A large centre island provides more storage with a stainless steel worktop, inset sink with mixer tap and electric hob. There is a range of appliances which may be available to any incoming purchaser.
There is a dining are within the kitchen with a large glass wall and roof providing an abundance of light and a door leading to the rear patio
Double doors lead to the utility/second kitchen with a range of units, a stainless steel sink with mixer tap, oven and four ring gas hob and housing for washing machine and tumble dryer. Door leading to the double garage.
Garage - A double garage with electric up and over door.
First Floor -
Master Suite - A fantastic master suite with a good sized bedroom area with built in wardrobes to one end of the room, a large window overlooks the front of the property with a glazed door leading to the balcony. There is an excellent en suite with low level wc, panelled bath with mixer tap and pedestal wash basin.
The master suite also has access to the drawing room via a spiral staircase.
Bedroom Two - A good sized double bedroom with a large window to the rear elevation and a range of fitted wardrobes.
Bedroom Three - An excellent double bedroom with large window to the rear elevation and a range of fitted wardrobes.
Bedroom Four - A large double bedroom with fitted wardrobes and a window to the rear elevation. This bedroom also benefits from an en suite comprising a wall hung wc, wash basin with storage unit beneath, tiled bath with mixer tap and shower head over. Floor to ceiling tiled walls and an opaque window to the front elevation.
Bedroom Five - A great sized double bedroom with a range of fitted furniture and window to the front elevation.
Family Bathroom - The family bathroom comprises a wall hung wc, wash basin with storage unit beneath, tiled bath with mixer tap and shower head over. Floor to ceiling tiled walls.
Externally - The property is accessed through wooden electric gates and a long tarmacked driveway leading to the front door and double garage.
The gardens are beautifully landscaped with mature shrubs and trees to the boarders providing privacy. There are flagged patio areas to the rear and side of the property as well as an extensive lawned garden area to the front.
This property is truly unique and needs to be seen to be fully appreciated. A MUST SEE!!!
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Disclaimer - Property reference 26508321. The information displayed about this property comprises a property advertisement. Rightmove.co.uk makes no warranty as to the accuracy or completeness of the advertisement or any linked or associated information, and Rightmove has no control over the content. This property advertisement does not constitute property particulars. The information is provided and maintained by Hibbert Homes, Hale. Please contact the selling agent or developer directly to obtain any information which may be available under the terms of The Energy Performance of Buildings (Certificates and Inspections) (England and Wales) Regulations 2007 or the Home Report if in relation to a residential property in Scotland.
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microRNAs (miRNAs) comprise an extensive class of post-transcriptional regulatory molecules in higher eukaryotes. Intensive research in Drosophila has revealed that miRNAs control myriad developmental and physiological processes. Interestingly, several of the best-studied miRNAs impact multiple biological processes, often by regulating distinct key target genes in each setting. Here we discuss the roles of some of these pleiotropic miRNAs, and their implications for studying and interpreting the roles of miRNAs in gene regulatory networks.
microRNAs (miRNAs) are an ancient class of genes that are abundant in higher eukaryotic genomes. Most animal miRNAs derive from primary transcripts bearing hairpin structures that are sequentially cleaved by the Drosha and Dicer RNase III enzymes to produce mature miRNAs (Ghildiyal and Zamore, 2009; Kim et al., 2009). In addition, a subclass of miRNAs known as mirtrons exploit the splicing machinery to generate their precursor hairpins prior to Dicer cleavage (Okamura et al., 2007; Ruby et al., 2007a). In either case, mature miRNAs are loaded into Argonaute complexes and guide them to target transcripts, generally leading to mRNA destabilisation, translational inhibition, or both (Liu, 2008).
Most characterized miRNA-target interactions include Watson-Crick pairing between the miRNA "seed" (positions 2–8 relative to its 5' end) and 3' untranslated regions (3' UTRs) of target mRNAs, although alternative pairing configurations and site locations have been documented (Bartel, 2009; Brennecke et al., 2005; Lai, 2002). On the basis of conserved seed matches in multigenome alignments, individual miRNAs are frequently predicted to target hundreds of transcripts (Krek et al., 2005; Lewis et al., 2005; Ruby et al., 2007b; Xie et al., 2005) (Figure 1A). Microarray and proteomic studies confirm the repression of large target cohorts by individual miRNAs (Baek et al., 2008; Lim et al., 2005; Selbach et al., 2008), although in most cases the consequence of miRNA-mediated regulation is quantitatively subtle.
Amongst the small population of transcripts bearing few predicted miRNA binding sites, gene ontology analysis reveals enrichment in basic cell metabolic processes such as DNA, RNA and protein synthesis (Farh et al., 2005; Stark et al., 2005). It may be generally undesirable for the activity of such "antitargets" to be hindered by miRNAs (Bartel and Chen, 2004), and these transcript categories are marked by shorter 3' UTRs and lowered density of predicted miRNA binding sites (Stark et al., 2005). On the other hand, as most transcripts are proposed to be miRNA targets (Friedman et al., 2009), most biological processes are plausibly under miRNA control in some form or fashion. By extension, one may infer that miRNA dysfunction has substantial possibilities to underlie human disease and cancer, and there is indeed a rapidly growing literature to support this notion (Croce, 2009; O'Connell et al., 2010; Williams et al., 2009).
Nevertheless, the all-encompassing nature of the proposed miRNA regulatory network has presented clear challenges for understanding the endogenous influence and importance of miRNAs. One rationalization of the large numbers of targets has been that miRNAs act as robustness factors for gene networks and gene expression patterns. For example, several cell- or tissue-specific miRNAs exhibit strong statistical enrichment for targets that are present at low levels, or are undetectable, within their expression domains (Farh et al., 2005; Sood et al., 2006; Stark et al., 2005). Largely mutually exclusive patterns of miRNA and targets led to the notion that miRNAs refine and/or reinforce gene expression patterns imposed by transcriptional regulation, perhaps to prevent spurious gene activity or to clear away earlier programs activated in progenitor cells. In this scenario, the role of miRNAs is secondary to transcriptional programs, and in select cases, comparison with transcriptional reporters suggested that miRNA activity was not the cause per se of non-overlapping miRNA and target accumulation (Stark et al., 2005).
On the other hand, genetic studies have also shown that certain miRNAs play critical instructive roles (Lee et al., 1993; Reinhart et al., 2000; Wightman et al., 1993), indicating that miRNAs are not relegated to cleanup or fine-tuning roles. It is also relevant to note that while heterozygosity or duplication of most genes is well-tolerated (Lindsley et al., 1972), minor changes in specific genes can be of substantial consequence to organismal phenotype. Taken together, these facts suggest that the deregulation of certain targets, amongst large cohorts of target genes, might be primarily responsible for observed phenotypes. Indeed, while genome-wide computational and experimental methodologies clearly reveal vast networks of miRNA:target interactions, there are now many documented cases for which the loss of individual miRNA-targeting events can largely account for phenotypes of miRNA gene mutants (Bushati and Cohen, 2007; Flynt and Lai, 2008) (Figure 1B).
Resolving these genomic and genetic viewpoints to more fully understand the biological functions of miRNAs requires additional research, particularly from in vivo contexts. Drosophila melanogaster is one of the most intensively studied model organisms, and has ~150 miRNA genes that are predicted to target >30% of its mRNAs via conserved binding sites (Ruby et al., 2007b). Previous reviews have covered many aspects of miRNA biology in this organism (Bushati and Cohen, 2007; Flynt and Lai, 2008; Smibert and Lai, 2008), and we do not intend this discussion to be inclusive of all the Drosophila literature. Instead, we focus on lessons and principles that emerge from in-depth analyses of four miRNAs: miR-7, miR-8, miR-9a and bantam. In particular, this work has led to a growing appreciation that (1) certain miRNA targets are of particular importance for organismal phenotype (Figure 1B), and that (2) individual miRNAs can have pleiotropic functions involving the regulation of different key targets in different developmental or physiological contexts (Figure 1C).
The mature miRNA produced by the mir-7 gene is completely conserved from annelids to mammals, indicating that it is an ancient miRNA (Lagos-Quintana et al., 2001; Prochnik et al., 2007). In fact, conserved putative regulatory sites corresponding to miR-7 binding sites were identified over a dozen years ago amongst a cohort of Drosophila genes that control neurogenesis (Lai and Posakony, 1997, 1998; Leviten et al., 1997). These include two large arrays of Notch target genes located in the Bearded-Complex (Brd-C) and the Enhancer of split-Complex [E(spl)-C], encoding multiple Bearded-family and basic helix-loop-helix (bHLH) repressor proteins. Indeed, ectopic activation of miR-7 in the animal yields specific phenotypes that phenocopy a reduction of Notch signaling during wing margin specification and peripheral nervous system development (Lai et al., 2005; Stark et al., 2003), consistent with the notion that these represent substantial in vivo targets of miR-7.
Initial studies of a Drosophila mir-7 deletion allele (that also removes portions of host and neighboring genes) revealed a reciprocal negative feedback loop between miR-7 and the transcription factor Yan during eye development (Li and Carthew, 2005) (Figure 2A). Yan is a neural repressor that is predominantly expressed in the undifferentiated portion of the retina, and it inhibits the transcription of mir-7 directly. Conversely, miR-7 is activated in specified photoreceptors, and directly represses yan via its 3' UTR (Li and Carthew, 2005). These regulatory interactions help to establish their mutually exclusive domains of activity.
Following this work, the Carthew group expanded these regulatory principles by showing that Yan and miR-7 both participate in coherent feedforward loops (FFLs) (Li et al., 2009). Coherent FFLs direct their desired output by both direct and indirect means, thereby ensuring robustness of outcome. In this setting, the Pnt-P1 transcription factor directly represses yan transcription and activates mir-7 transcription, the latter of which represses yan post-transcriptionally. Reciprocally, Yan represses the transcription of both mir-7 and phyllopod, the latter of which encodes an E3 ubiquitin ligase that degrades TTK69, also a repressor of mir-7 transcription. In short, Yan represses mir-7 transcription through both direct and indirect mechanisms. The interlocking of these coherent FFLs generates the reciprocal negative feedback loop between miR-7 and Yan, which in turn helps to coordinate the orderly specification of photoreceptors.
miR-7 also regulates the development of proprioceptive (chordotonal) organs (Li et al., 2009) (Figure 2B), and E(spl)-C targets are relevant in this context (Lai et al., 2005; Stark et al., 2003). In contrast to retinal development, however, miR-7 participates with E(spl)-C and the proneural transcription factor Atonal in an incoherent FFL in this context (Li et al., 2009). In this schema, an upstream factor activates two targets with opposing functions. In particular, Atonal directly activates the transcription of many E(spl)-C genes as well as of mir-7, which as mentioned represses multiple E(spl)-C genes. These genes are further connected as E(spl)-bHLH repressor proteins inhibit atonal transcription. In such an incoherent FFL configuration, fluctuating Ato levels may result in back and forth repression between Atonal and E(spl)-C genes, but robust Atonal expression will result in sustained repression of E(spl)-C genes via miR-7, further strengthening Atonal expression. The objective and outcome of these complex gene regulatory networks is to permit neural specification only in response to strong Atonal activity.
It is relevant to bear in mind that miR-7, in stark contrast to Yan, Ato, and the E(spl)-C, is mostly dispensable for eye and chordotonal organ development under standard laboratory conditions (Li and Carthew, 2005). Instead, it was proposed that many regulatory connections of miR-7 in feed-forward and feedback loops provide robustness to the precise nature of nervous system specification. To test this theory, mir-7 mutants were subjected to environmental perturbation using temperature cycling. Indeed, the combination of mir-7 deletion and fluctuating temperature resulted in reduction of Ato, increased and irregular accumulation of Yan, and defects in eye and sensory organ development (Li et al., 2009).
A distinct role for miR-7 in the differentiation of germline stem cells (GSCs) was uncovered through analysis of mutants in maelstrom (mael), which encodes an HMG-box transcription factor (Findley et al., 2003; Pek et al., 2009). In wild-type testes, GSCs maintain contact with the somatic hub and divide asymmetrically to produce two cells; a differentiating gonialblast (GB), which loses contact with the hub, and the stem cell, which retains contact with the hub. In male flies lacking mael, differentiation fails in the cells that should have become the GBs, leading to strong accumulation of GSC-like cells in the testes and a decline in fertility with age.
This phenotype is reminiscent of loss of function mutations in bag of marbles (bam), a novel gene that regulates GSC progression, and Bam is in fact reduced at the post-transcriptional level in mael testes (Pek et al., 2009). The bam 3' UTR contains binding sites for several miRNAs, including miR-7, and a bam 3' UTR sensor was sensitive to miR-7 dosage in vivo. A regulatory cascade was extended by the observation that miR-7 levels are increased in mael testes, concomitant with reduction of a repressive histone mark at the mir-7 locus. Since Mael binds to consensus HMG binding sites upstream of the mir-7 promoter, Mael may directly repress mir-7 during GB differentiation. The significance of miR-7 upregulation in mael mutants was shown by the finding that heterozygosity for mir-7 partially suppressed the differentiation defect in mael testes (Figure 2C). Recently, miR-7 was also implicated in the development of ovarian GSCs through the regulation of the cyclin-dependent kinase inhibitor dacapo (Figure 2D), providing evidence for roles of miR-7 in both the male and female germline (Yu et al., 2009).
Altogether, these studies of miR-7 introduce the concept that a single miRNA can have different key targets in different developmental contexts. However, while miR-7 is identical between flies and humans, these data are not necessarily informative of key miR-7 targets in vertebrates. Indeed, while many miRNA targets can be defined on the basis of their evolutionary conservation amongst flies, amongst nematodes, or amongst vertebrates, there turn out to be very few targets shared by miRNAs that are conserved in all three of these species clades (Chen and Rajewsky, 2006). This has led to the notion that miRNA targets are acquired and lost quite frequently across large evolutionary distances. Indeed, vertebrate orthologues of yan and E(spl)bHLH genes do not contain miR-7 sites that are conserved amongst vertebrates, and miR-7 is not apparently expressed in vertebrate chordotonal organs (Li et al., 2009). In the case of the GSC requirement for miR-7, despite the evidence of a functional role for miR-7 in repressing Bam expression in GBs, the binding site analyzed is not even present in the closest sister species to D. melanogaster, namely D. simulans and D. sechellia (Pek et al., 2009). These results suggest that mRNAs can readily evolve new regulatory relationships with conserved miRNAs.
Another miRNA well conserved through animal evolution is miR-8. There is a single copy of mir-8 in flies, but mammalian genomes have 5 paralogues in two distinct genomic clusters that are known as the miR-200 family. Interestingly, in contrast to the studies to date with miR-7, several important miR-8 targets and biological activities appear to be conserved between invertebrates and vertebrates.
The first study of miR-8 loss of function reported flies with reduced survival, malformed legs and progressive neurodegeneration (Karres et al., 2007) (Figure 3A). As with most other miRNAs, there are many (in this case perhaps >300) targets of miR-8 that are predicted to be broadly conserved amongst the Drosophilids (e.g., http://www.targetscan.org/). If the derepression of all of these contributed equivalently to the mir-8 mutant phenotype, then it would be unlikely that any particular gene would emerge as a key target responsible for phenotype. Amongst genes upregulated in mir-8 mutants, the transcriptional corepressor encoded by atrophin has several miR-8 binding sites in its 3' UTR and was responsive to miR-8 in sensor tests. Remarkably, ectopic expression of atrophin in miR-8-expressing cells phenocopied mir-8 mutants, while heterozygosity for atrophin could partially rescue the survival and leg defects of mir-8 mutants. These data therefore support a particularly substantial role for miR-8-mediated repression of atrophin, out of the hundreds of genes that bear conserved miR-8 binding sites. Notably, the human ortholog of atrophin, RERE, has three potential binding sites for mammalian miR-8 members, and is responsive to their activity in cultured cell assays (Karres et al., 2007). Therefore, this regulatory linkage may be deeply conserved across animals.
An overexpression screen for negative regulators of the Wingless (Wg)/Wnt pathway in Drosophila also identified miR-8 (Kennell et al., 2008). In this role, miR-8 directly represses wntless (wls), a gene required for export of the Wg morphogen (Figure 3B). Overexpression of miR-8 in vivo decreased levels of Wls cell-autonomously, with a concomitant decrease of extracellular Wg. A direct regulatory relationship with miR-8 was also reported for the positive regulator of the Wg pathway CG32767 (DasGupta et al., 2005; Kennell et al., 2008), suggesting that miR-8 negatively regulates the Wg pathway at multiple levels. Using a mouse cell line in which inhibition of Wnt signalling leads to adipogenesis, ectopic expression of mammalian miR-200 family members also induced adipogenesis marker genes and increased lipid accumulation. Although the endogenous influence of these miRNAs on signaling remains to be demonstrated, these studies suggest that a role for miR-8 in repressing Wg/Wnt signaling may be conserved in mammals.
Identification of miR-200 family miRNAs as growth promoting factors when ectopically expressed in human cell culture led to re-analysis of the original mir-8 mutant flies (Hyun et al., 2009; Park et al., 2009). This uncovered a significant growth defect at larval stages, slightly delayed eclosion and reduced size in adults for flies lacking miR-8 (Figure 3C). This appeared to be due to reduced cell number, with no obvious difference in cell size. In addition to previously reported expression patterns (Karres et al., 2007), mature miR-8 was strongly expressed in the fat body, and fat body-specific expression of miR-8 was sufficient to rescue the size defect of mir-8 mutants (Hyun et al., 2009). Notably, neuronal expression of miR-8 did not rescue the growth defect, demonstrating that the growth and neuronal phenotypes of mir-8 mutants are distinct.
To identify the relevant target(s) of miR-8 in the fat body, predicted targets of Drosophila miR-8 and the human miR-200 family were pooled from multiple computational schema, and mined for shared targeting of orthologous transcripts. Out of 15 orthologous pairs, seven were found responsive to their cognate miRNAs in their respective cell assays. Fat body-specific knockdown of these candidates using inverted repeat transgenes was used to assess whether reduction of any of them could rescue the small body phenotype of mir-8 mutants. Interestingly, depletion of the zinc finger protein encoded by u-shaped (ush) rescued mir-8 flies to wild-type weight. Notably, ush knockdown did not increase the weight of wild-type flies, indicating that the rescue is not simply an additive effect but is specific to the mir-8 mutant condition. Further analysis showed that Ush is elevated in mir-8 larvae, and that Ush and its human orthologue FOG2 regulate the PI3 kinase pathway by directly interacting with the regulatory subunit of PI3 kinase (Hyun et al., 2009).
Amongst the varied phenotypes associated with manipulation of miR-8, the most recent finding is its control over neuromuscular junction (NMJ) formation (Loya et al., 2009). In flies null for mir-8, NMJs are smaller, less branched, and have fewer boutons than in wild-type (Figure 3D). This phenotype was recapitulated by ubiquitous expression of a miR-8 transgenic sponge, a decoy target that acts as a sequence-specific competitive inhibitor of miRNA function (Ebert et al., 2007). Transgenic knockdown of miRNA activity permitted a test of whether miR-8 was required pre- or post-synaptically. In fact, the mir-8 mutant phenotype was obtained when the sponge was expressed post-synaptically in the target muscles, but not when expressed in the neurons. These phenotypes resembled manipulation of Enabled, an EVH1/PH domain regulator of synapse development. Indeed, enabled is a direct miR-8 target whose protein output is substantially increased in mir-8 mutants. More to the point, reduction of enabled dosage, either systemically or specifically in the muscle, detectably alleviated mir-8 mutant and miR-8-sponge phenotypes. These data provide strong support that deregulation of Enabled in muscles is causative for the NMJ phenotype of mir-8 mutants.
The collected analyses of miR-8 provide a curious contrast to those with miR-7. Both miRNAs are conserved between invertebrates and vertebrates, but the major targets of Drosophila miR-7 to date (yan, multiple E(spl)-C and Brd-C genes, bam) are not similarly conserved. In contrast, the orthologs of two different major confirmed targets of Drosophila miR-8 (atrophin and ush) were reported as targets of the orthologous miR-200 family in vertebrates, and there is some evidence suggesting that miR-8 represses Wg signalling in both flies and mammals. The meta-analysis of combined outputs from many prediction algorithms yielded only a handful of targets for miR-7 (Li et al., 2009) and miR-8/miR-200 (Hyun et al., 2009) that are even conceivably shared between mammals and flies (9 for miR-7, 15 for miR-8/miR-200), in agreement with previous conclusions on a paucity of deeply conserved miRNA:target reglationships (Chen and Rajewsky, 2006). The analysis of such deeply conserved targets of miR-7 may conceivably reveal important biological interactions, as should investigation of additional clade-specific targets of miR-8 (such as enabled).
Drosophila miR-9a was originally described as a gene that exhibited strong target avoidance in its expression domain (Stark et al., 2005). In the early embryo, mir-9a is specifically expressed in the presumptive ectoderm and neuroectoderm, but is later maintained only in the ectoderm and is excluded from neural tissue. mRNAs with the ontological classification "neurogenesis" are enriched for the shared target sites of the related miR-9a/b/c miRNAs, suggesting that expression of miR-9 family miRNAs is mutually exclusive to many of its targets. It was inferred that a role for miR-9a is to suppress the neural potential of ectodermal derivatives (Stark et al., 2005).
Flies lacking miR-9a exhibit a mild ectopic sensory organ phenotype in the larva, on the anterior wing margin, and on the thorax, consistent with a failure to correctly repress neural identity (Li et al., 2006) (Figure 4A). These flies also exhibited a completely penetrant loss of posterior wing margin. Here, the zinc finger proneural gene senseless (sens) was reported as a critical target of miR-9a. In particular, heterozygosity for sens restored the correct number of sensory organs on the anterior wing margin and could partially rescue the loss of posterior wing margin in mir-9a mutants (Li et al., 2006). Based on the ectopic but variable increase of sensory organs in mir-9a mutant, it was proposed that miR-9a helps prevent in appropriate SOP specification by keeping sens levels below a threshold needed to lock in the SOP fate robustly (Cohen et al., 2006; Li et al., 2006). Therefore, sens appears to be one of the key neurogenesis targets of miR-9a, amongst a cohort of neural targets of this miRNA.
Although the strong wing defect of the mir-9a mutant was reported earlier (Li et al., 2006), its developmental basis was not examined. Recent studies showed that mir-9a mutant wing pouches, assayed during larval development, showed defects in wing margin gene expression, broad ectopic apoptosis, and upregulation of the transcriptional regulator Drosophila LIM-only (dLMO) (Bejarano et al., 2009; Biryukova et al., 2009) (Figure 4B). Indeed, dLMO proved to the critical target of miR-9a in this setting. In contrast to heterozygosity for sens, removal of one copy of dLMO strongly reduced ectopic apoptosis during wing development and completely rescued the loss of posterior margin seen in mir-9a mutant flies (Bejarano et al., 2009; Biryukova et al., 2009). A further twist is that dLMO has an additional pre-pattern function to specify certain proneural territories (Asmar et al., 2008); in this context, it acts developmentally upstream of Sens. Heterozygosity for dLMO can partially suppress the ectopic sensory bristles present on the notum of mir-9a mutants, although not to the same extent as the reduction of sens (Bejarano et al., 2009). Therefore targeting of both Sens and dLMO by miR-9a are relevant to both PNS and wing development, but Sens may be more critical in the PNS and dLMO more critical in the wing.
In retrospect, the realization of a profound requirement for regulation of dLMO by miR-9a was the culmination of nearly 100 years of Drosophila genetics. dLMO was hit genetically in the 1920s by dominant Beadex mutations that induced wing loss (Morgan, 1925) similar to mir-9a mutants, and these proved to be transposon insertions that disrupted the 3' UTR of dLMO (Milán et al., 1998; Shoresh et al., 1998; Zeng et al., 1998). In fact, dLMO levels must be so carefully controlled that even 2x dosage resulting from duplication of the dLMO locus can induce the Beadex mutant phenotype, i.e. wing margin loss (Green, 1952; Lifschytz and Green, 1979). This history serves to remind us that while the general appreciation of the importance of miRNA regulation has come quite recently, the genetic evidence for important miRNA targets has been with us for much, much longer.
Interestingly, miR-9a appears to have quite different relationships with its two best-characterised targets. With sens, the thresholding model suggests that before cell fate decisions have been made, miR-9a and sens are co-expressed in the same cells, where miR-9a acts to keep sens levels below a threshold that would trigger SOP formation without other appropriate signalling events. Once SOPs are defined, the expression patterns become mutually exclusive. The relationship with dLMO appears quite different. miR-9a is detected by in-situ hybridisation throughout the entire wing imaginal disc (Biryukova et al., 2009; Li et al., 2006), and dLMO protein is similarly quite broadly expressed (Milán et al., 1998). In addition, clonal loss of mir-9a leads to upregulation of both dLMO and a GFP sensor for miR-9a activity throughout the whole wing disc (Bejarano et al., 2009), indicating that miR-9a is expressed and acts in the same tissues as one of its primary targets. This is more indicative of a tuning relationship than a mutually exclusive relationship. Although the thresholding model also requires co-expression of miRNA and target, once the threshold is exceeded, the expression patterns are expected to become mutually exclusive. Therefore, the same miRNA may act in different manners with different targets in different tissues.
The three previous examples concerned miRNAs that are deeply conserved between invertebrates and vertebrates. Even so, some of the phenotypes that have been characterized might arguably be considered subtle, and dependent on the expertise of particular laboratories for their elucidation. For example, the notum peripheral nervous system defect of mir-9a mutants comprises an extra bristle or two in only a subset of animals, whereas the eye phenotype of mir-7 mutants is only revealed by sensitized genetic backgrounds or environmental perturbations. Since miRNAs that are more deeply conserved appear to generally have more targets than those that are more recently evolved, one might suspect that miRNAs with lesser evolutionary range might correspondingly be of more subtle biological importance. However, this is clearly not the case. For instance, miR-279 is insect-restricted but is mostly lethal prior to adult hatching, and causes a strong defect in olfactory nervous system specification (Cayirlioglu et al., 2008). miR-iab-8 is similarly insect-restricted, but its loss results in complete sterility in both sexes (Bender, 2008).
Perhaps the most compelling example of a Drosophila miRNA with dramatic biological functions, but lacking vertebrate homologs, is that of bantam. Amongst the first miRNAs to be characterized genetically, bantam was initially defined as an enigmatic locus that regulated growth (Hipfner et al., 2002; Raisin et al., 2003) and only later appreciated to encode a miRNA (Brennecke et al., 2003). Ectopic bantam promotes overgrowth of various tissues such as eyes and wings, while flies carrying hypomorphic alleles of bantam are small. The complete loss of bantam is lethal at larval and pupal stages, and clonal analysis of tissues homozygous for a null allele revealed their inability to proliferate substantially (Figure 5A). The pro-apoptotic gene head involution defective (hid) was a particularly compelling target of bantam, with 5 conserved target sites in its 3' UTR (Brennecke et al., 2003). Interestingly, suppression of apoptosis does not induce overt tissue overgrowth, as seen with bantam overexpression, suggesting that additional growth targets of bantam exist. These inferred targets remain elusive to date.
The first evidence for regulation of bantam by pathways that promote cell growth was the ability of bantam to modify the small, blistered eye phenotype of ectopic expression of the tumour suppressor expanded (Hipfner et al., 2002). Co-expression of bantam suppressed the phenotype resulting in almost wild-type morphology, whereas loss of one copy of bantam in this background resulted in a smaller eye with more pronounced blistering (Hipfner et al., 2002). Subsequently, Expanded was shown to be a key apical component of the Salvador/Warts/Hippo (SWH) growth control pathway (Cho et al., 2006; Hamaratoglu et al., 2006; Silva et al., 2006), for which loss-of-function of various cytoplasmic and transmembrane components results in massive tissue overgrowth and resistance to apoptosis.
The phenotypic similarity of bantam gain-of-function and loss-of-function of various SWH pathway components raised the question of whether bantam was a transcriptional target of the SWH pathway. Indeed, bantam proved to be a target of the Yorkie transcriptional co-activator (Nolo et al., 2006; Thompson and Cohen, 2006), whose nuclear function is suppressed by activity of the SWH pathway (Huang et al., 2005). Accordingly, overexpression of Yorkie in clones mutant for bantam failed to yield overgrowth, suggesting that bantam is required for Yorkie-induced overgrowth. Conversely, forced activation of bantam in cells lacking yorkie resulted in overgrowth, suggesting that ban is a key growth-promoting transcriptional target of Yorkie (Nolo et al., 2006; Thompson and Cohen, 2006). More recently, bantam was shown to be a crucial downstream target of the SWH pathway in the proliferating cells of the anterior eye imaginal disc, where Yorkie complexes with different transcription factors compared to the wing (Peng et al., 2009). Therefore, bantam is a crucial target of growth regulatory pathways in a variety of cellular and developmental contexts.
The most well characterised bantam:target interaction is with the pro-apoptotic gene hid (Brennecke et al., 2003; Nahvi et al., 2009), but only recently has this been analysed in the context of endogenous levels of the two genes (Figure 5B). In addition to their growth defects (Brennecke et al., 2003), bantam mutants exhibit ectopic apoptosis, and animals homozygous for hypomorphic alleles of bantam also show significantly increased levels of apoptosis in response to ionizing radiation (IR) (Jaklevic et al., 2008). Bantam also prevents rbf mutant interommatidial cells from undergoing cell death in the developing eye disc (Tanaka-Matakatsu et al., 2009), demonstrating that cell autonomous anti-apoptotic activity mediated by bantam is also present in the eye disc.
Consistent with a role in limiting excess apoptosis in response to IR, bantam activity is strongly increased soon after IR, as measured by decreased levels of a bantam-responsive sensor, however, this does not appear to correspond to significant changes in levels of mature bantam (Jaklevic et al., 2008). Suppression of apoptosis in irradiated cells by expression of the baculovirus inhibitor of apoptosis protein p35 prevents the downregulation of the bantam sensor, suggesting that the upregulation of bantam activity is due to a signal from apoptotic cells and is not a direct response to DNA damage. p53, which is a crucial gene in DNA damage response, is required for the increase in bantam activity induced by IR (Jaklevic et al., 2008). On the other hand, IR exposure is also expected to result in p53-mediated activation of the SWH pathway activation, which is expected to decrease bantam transcription leading to apoptosis (Colombani et al., 2006). These apparently contradictory data may be reconciled by the non-autonomous role of apoptosis in inducing bantam activity. Perhaps p53 is inducing SWH pathway activation in cells that have sustained DNA damage beyond a certain threshold, leading to apoptosis and a growth-promoting signal to neighbouring cells, which then increase their bantam activity. This may explain the absence of an obvious increase in bantam levels in these irradiated discs.
Animals that are hypomorphic for bantam exhibited defects in GSC maintenance, with most animals having no remaining GSCs after 14 days (Yang et al., 2009) (Figure 5C). Furthermore, clones of larval primordial germ cells (PGCs) that are null for bantam showed branched fusome structures, which are markers of PGC differentiation, suggesting that bantam limits PGC differentiation. Interestingly, in this study, GSC clones of bantam mutants did not exhibit any significant defects in maintenance, suggesting that the requirement for bantam in GSC maintenance is non-autonomous and may potentially reside in the GSC niche (Yang et al., 2009). Curiously, a separate study described a clear phenotype of defective maintenance for bantam mutant GSC clones (Shcherbata et al., 2006). Both studies utilized similar techniques involving null deletion alleles for bantam, so the reason for this discrepancy is unclear. A separate study demonstrated that the bantam sensor is strongly downregulated in GSCs compared to 16 cell cysts, suggesting that bantam is active in GSCs and inactive in the differentiating cysts (Neumuller et al., 2008).
Very recent studies of bantam revealed its unexpected influence on the differentiation of post-mitotic neurons. In the early Drosophila larva, sensory neurons tile across the body wall to create a sensory field. As the larva grows, a fixed number of sensory neurons must expand their fields of coverage by extending dendrites in proportion to the growth of the organism as a whole, a process referred to as scaling growth. Amongst a panel of mutants that exhibit defective larval growth, animals null for bantam established normal coverage of the receptive field in early larvae, but failed to attenuate scaling growth after cessation of larval growth (Parrish et al., 2009) (Figure 5D). Strikingly, this phenotype was rescued by expression of bantam in the overlying epithelial cells, and not in the sensory neurons, demonstrating that bantam is required in these epithelial cells to provide a signal that attenuates scaling growth of sensory neurons (Parrish et al, 2009). Therefore, as hinted at by studies in GSCs, bantam also has important non-autonomous roles during dendrite tiling.
A distinct role for bantam in neural function was revealed during studies of the circadian clock. The eponymous clock gene encodes a bHLH-PAS transcription factor that is a central generator of circadian rhythms, and is also a substantial target of bantam (Kadener et al., 2009). Overexpression of bantam in timeless-expressing cells, which comprise the central pacemaker, resulted in lengthening of the circadian period by three hours (Figure 5E). This phenotype is similar to the periodicity phenotype caused by loss of clock function, and the annotated clock 3' UTR is indeed predicted to have a single bantam binding site. However, further studies revealed that the predominantly expressed clock 3' UTR is substantially longer and bears three functional bantam binding sites, which are responsive to bantam repression in cultured cell assays. This provides a motivation to be cautious in assessing computational target predictions, which do not necessarily account for all in vivo transcript isoforms.
Unlike other characterized settings of bantam function, it is technically challenging to remove bantam specifically from pacemaker cells to check for a circadian phenotype. Still, an informative test can be done in cis to assess the functional impact of bantam sites in the clock transcript. Compared to a wild-type clock transgene, a clock transgene bearing specific mutations in its bantam binding sites was unable to provide full genetic rescue of the clock null mutant background (Kadener et al., 2009). This constituted compelling evidence for the endogenous regulation of clock by bantam to generate normal circadian rhythm.
The wide range of severe phenotypes resulting from loss of bantam are notable in light of the widely-held view that miRNAs are primarily fine tuning factors. Through regulation of its so far described targets hid and clock, bantam regulates processes as diverse as apoptosis and circadian rhythm. Since bantam has additional significant roles in promoting growth, maintaining GSCs and non-autonomously attenuating dendrite scaling, it will be interesting to see what other key targets it has, and whether bantam functions via "a few" or "many" targets in these various settings.
Through the analysis of miRNA mutants and the regulatory relationships they reveal with target genes, we are beginning to understand more about the complex and varied roles of miRNAs in animal development. The collected studies in the Drosophila model affirm the conclusion that there is no single rule or reason for miRNA regulation of a given process.
miRNAs undoubtedly have many, perhaps hundreds of direct targets, and bioinformatic studies reveal underlying logic to these networks. One conclusion is that miRNAs may suppress broad transcript cohorts that are leftover from progenitor states, or that may arise spuriously because of alternative developmental potentials of a given celltype. In this view, miRNA regulation is secondary to transcriptional regulation and serves as a general backup to reinforce a particular cell state. If the individual regulation of many targets is not critical for gross phenotypes, this might permit a great deal of evolutionary fluidity in the target networks for otherwise highly conserved miRNAs. On the other hand, there is also abundant evidence that miRNAs can have key target genes that explain a great deal about the genetic function of the miRNA gene. And at least some of these cases require the function of the miRNA in target-expressing cells, so that miRNAs play instructive roles to deliver just the right amount of target activity.
Although some of key miRNA targets were revealed as "top" predictions that contain multiple binding sites for the miRNA (e.g. hid with 5 highly conserved binding sites for bantam), other key target genes have only single target sites (e.g. dLMO with only one clear site for miR-9a) or have target sites that did not exist in annotated 3' UTRs (e.g. clock with seemingly downstream bantam sites). For many functional analyses of miRNAs, then, genetic phenotypes were critical for understanding the role of the miRNA and providing a rationale to pursue individual targets in depth.
Early evidence for the breadth of the miRNA target network came from focusing on conserved targets, and certainly conservation implies a likelihood of beneficial function. However, highly conserved features do not necessarily imply that they mediate processes that yield obvious phenotypes in the mutant situation. Certainly there are many highly conserved putative transcriptional regulatory regions for which no activity could be defined in vivo, and there are many highly conserved protein-coding genes whose deletion similarly causes no obvious defect. Nevertheless, small quantitative deviations in gene activity can result in dramatic costs to fitness, especially when measured not in a few generations in optimal laboratory conditions, but instead across evolutionary scales of time and on the population level. Therefore, the consequences of deleting some conserved miRNA genes, and potentially of removing many conserved miRNA binding sites, may need to be studied under challenging environmental conditions, under sensitized genetic backgrounds, or using highly quantitative strategies.
On the other hand, the simple lack of deep conservation may not be taken as strict evidence for lack of useful functions. We have discussed that the knockouts of several miRNAs that are conserved between invertebrates and vertebrates have relatively modest effects in Drosophila, whereas the invertebrate-specific miRNA bantam plays crucial roles in a dazzling variety of developmental and physiological settings. bantam is essential in both proliferating and post-mitotic tissues, and regulates both cell-autonomous and non-cell autonomous processes. Other insect-specific miRNAs seem to have crucial functions even as measured by such crude measures as viability and fertility. Going to the more "micro" level, there are now cases of newly-evolved miRNA:target relationships that appear to play important roles in specific Drosophilid species. Perhaps changing miRNA:target relationships have played active roles in creating species-specific characteristics.
Finally, we are fascinated by the fact that the continued study of individual Drosophila miRNA mutants continues to reveal additional phenotypes caused by the deregulation of newly defined target transcripts. Pleiotropic functions of miRNAs may provide a contributing rationale for the existence of extensive target networks for individual miRNAs. As is the case for many genes encoding transcription factors and other regulatory factors, we presume that there will continue to be more to miRNA activities than is revealed at first glance.
We thank Zhigang Jin for critical reading. Work in E.C.L.'s group was supported by the Bressler Scholars Fund, the Burroughs Wellcome Fund, and the NIH (R01-GM083300 and U01-HG004261).
Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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Biology is one of the original natural sciences, even reaching back as far as the study of anatomy in ancient Egypt, where evidence has been found of surgery, forensics, and anatomical knowledge of both organ structure and function. Biology in general is the study of life and living organisms, with specific attention to function, growth, evolution, identification and taxonomy. Modern biology is becoming an interdisciplinary field, overlapping with biomedical engineering, molecular chemistry, and genetics. The biological sciences recognize the cell as the basic unit of life, and DNA as the basic unit of heredity. Moreover, all organisms survive by consuming and transforming energy via respiration or photosynthesis, and by regulating their internal environment via homeostasis.
The life sciences are defined by the scale at which organisms are studied, in that biochemistry focuses on chemical processes of life while molecular biology involves molecules and protein synthesis. Cellular biology may be broken down further into a structural analysis of cell organelles and lipid membrane boundries, such as in skin tissue or alveolar sacs in the lungs. In the field of genetic engineering, DNA (deoxyribonucleic acid) may be spliced together from different organisms, in an attempt to transfer hereditary traits that may increase crop yields, create a supply of stem-cells, or even clone sheep. Beyond major requirements within the human anatomy department, you may consider Moraine Valley College clinical pharmacology as a complementary elective to your primary course of study.
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ATLANTA – Georgia has been named the nation's best state for business for the fourth year in a row by Site Selection magazine.
The announcement came during a press conference held Wednesday morning by Gov. Nathan Deal and other state officials.
Site Selection magazine is an economic development trade publication.
Each November, Site Selection releases rankings for the top states in which to do business. Georgia has been ranked among the top 10 states in Site Selection's business climate rankings throughout the last decade and has held the top spot for the last three years.
"In the last four years, many small businesses have expanded in Georgia and numerous international companies have established operations here, reflecting the foundation offered to job creators," Deal said. "Georgia leads the way in providing companies with a low tax burden and a business-friendly climate. In return, there have been more than 575,000 private sector jobs created since 2011."
Deal has made economic development a focus of his tenure as governor, vetoing several measures passed by recent General Assembly sessions that he deemed as damaging to the state's business-friendly image. Earlier this year, the legislature passed a so-called religious liberty bill, which drew the ire of many major Hollywood, entertainment and sports-related businesses because of its perceived anti-gay bias.
Deal's veto raised the ire of conservative, religious leaders in the state who have vowed to revisit the issue when the legislature convenes in January 2017.
"Georgia's fourth consecutive top state business climate ranking is based on where corporate expansion projects are occurring and where investors say they want to commit their capital for the long term," said Mark Arend, editor-in-chief of Site Selection. "Our readers say the state's fiscal soundness, predictable economic and regulatory climate, workforce and transportation infrastructure are their main reasons for investing in Georgia."
Georgia top business state for 4th straight year
Georgia has been named the nation's best state for business for the fourth year in a row by Site Selection magazine.
ATLANTA – Georgia has been named the nation's best state for business for the fourth year in a row by Site Selection magazine.
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We've had four consecutive nights of very hard frost this week. I love to see the winter veg toughing it out in these conditions, there's something very comforting about their defiant attitude. Winter weather – who cares?
In the low light of a cold sunrise, the garden, orchard, field and wood have been a spectacular winter wonderland.
We've been toughing it out ourselves, too, as our boiler decided to break down – how typical is that in the coldest spell so far this year? Thank goodness for The Beast and our mighty log pile. We've been putting all that heat to good use, roasting trays of pumpkin to freeze for soup and, on a quick trip to the shops, I just couldn't resist adding yet another seed variety to my ever-growing squash pile for next year…
I can't believe how quickly we're motoring through our stored garlic, there's only one plait left in the shed. It was definitely the right decision to plant twice as much this year, pink and white varieties are both looking very happy.
I'm happy, too: the boiler is now fixed and the weather has turned very mild. In fact, we're forecast temperatures up in the teens over the next few days so time to get back to some digging. I've almost finished turning over the patch of ground down the side of the tunnel. This will be where the spuds go next year with plenty of room for earlies and maincrop varieties this time.
I've moved my trusty silage bags to cover a new area in front of the leeks, to kill the grass and be dug in spring. With some of the patch now planted with asparagus and globe artichokes for several years, I need to extend the planting area (well, that's my excuse, anyway – I'm not sure Roger is convinced) and this seems the logical place to start. All those squashes have got to go somewhere, after all.
I've also started a major revamp of the flower garden, poor neglected area that it is. Lots of digging needed here to extend it and create a large area of heat-loving, drought-resistant plants which can cope with the summer temperatures. I'm planning to use it as a separate extension of the veg patch, I think globe artichokes, climbing borlotti beans and rainbow chard will all add their own charm, maybe some tomatoes, too, to avoid the blight problem… in fact, will there actually be any room for flowers, I wonder?
The herbs I've grown from seed for this border and the veg garden are all tucked up in the tunnel in readiness for their new homes next spring.
I'm also planning to incorporate some plants for dyeing wool, starting with coreopsis, chamomile, weld and woad. The latter is a member of the cabbage family so I need to keep it well away from the veg patch – just hope it doesn't have all the problems my poor brassicas have had this year (are you listening, pigeons?). I know I'm unlikely to achieve the strong colours possible with synthetic dyes – this is a picture of the first skein of commercially-dyed Shetland wool inspired by autumn colours in my latest spinning project-
but natural methods have far more appeal to me. So what if the colours are more subtle, it makes the whole process more interesting. I should be able to use some vegetables, too – onions being top of the list.
Prize for the most confused plant in the garden this week goes to this little beauty!
On a more seasonal note, the frosts have really done for the leaves this week and the apple orchard suddenly looks bare. Now we can spot the mistletoe lurking in the branches, and despite the fact that we cleared lots out earlier in the year for the sake of the trees, there won't be any shortage when it comes to gathering some Yuletide greenery next week … and plenty of berries should mean lots of Christmas kisses! 🙂


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Companies – My Most Valuable Advice
Three Options for Business Phone Systems Business maintains telecommunication networks for the phone communication. There are different types of phone systems that can be used by companies. Both the business structures and features of the phone systems must be factored in when making decisions on selection of phone systems. The company has to decide whether it needs to have a dedicated phone system or use the mobile phones. In case the business requires a dedicated phone system, it will have to make a choice among three alternatives. The choice will depend on multiple factors including the number of employees who need to connect to the phone system. Virtual phone systems are among the choices and allow employees based in another area other than the company office to connect to the company office. When there is a call made to the main office telephone number; it is auto-redirected to the employee mobile of the home phone. It helps the organization maintain a professional outlook at a lower cost. It offers automatic call forwarding, faxing, voice mail and call screening.
Resources: 10 Mistakes that Most People Make
The landline phone system is another option. Copper wires are utilized in making the connections. The services are offered by landline telephone companies. An office based PBX hardware needs to be installed. It may require large upfront investments making it expensive for small business. Business with poor internet connections can find it very attractive. This option is fine for firms with large capital base since it may prove cheaper. The presence of an in-house support staff in called for maintenance.
Lessons Learned About Services
The VoIP phone system is the most recent phone system. Instead of using copper wires to make the connections, it uses the existing Internet connections the business has. Its applications are best for firms with high speed internet connections. VoIP phone systems are bale to connect to remotely located employees on their mobile phones. When a firm decided to use a VoIP system; it can apply self hosting or hosted services. Business buy and own the assets that are sued to run the self hosted VoIP system. Notable among these is the PBX hardware that maintains the circuit connections.Though expensive upfront, the system is cost effective in the long run. It requires that the firm has an internal staff who can maintain the system. It is upon the business to ensure that the system is running. Otherwise, they would have to call for external assistance which might b costly. A business which wants to be in total control of its phone communications should go for this option. Cloud technology is used for the hosted VoIP systems. Other than the phones, the business does not buy other assets. The telecommunication company takes care of all the facilities. The payment is made for each month based on the number of users.


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ZANESVILLE, Ohio-Â The Muskingum County Commissioners met Monday afternoon to talk about saving money.
The commissioners enlisted SpyGlass Group, LLC, to perform a telecommunications audit to ensure they are running at proper efficiency.
Jim Long, Director of Implementation and Project Management at SpyGlass Group, said the main goal was to see if they could help them save money.
"We ran a telecommunications audit for the county, where we gathered the invoices and put together inventories for them," said Long. "[It's] to make sure they're using what they are paying for and paying the lowest amount for what they are."
Long said that typically its protocol for the providers to increase the rates after a contract comes out of expiration.
"All of the funding for cities and counties is tax dollars," said Long. "Saving that money to get that back into the budget is obviously what the tax payers really want to see happen so they know its not just going to the big corporations of AT&T, Verizon [and] what have you."
SpyGlass Group also worked with other cities and counties in the area.


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Supplier Homepage Product Filteration Equipment Hot Selling Rotary Vacuum Drum Filter Press
Hot Selling Rotary Vacuum Drum Filter Press
|FOB Price:
|US $1-100,000 / Piece
|Min. Order:
|1 Piece
|Min. Order
|FOB Price
|1 Piece
|US $1-100,000/ Piece
|Qingdao, China
|Production Capacity:
|Payment Terms:
|L/C, T/T, Western Union, Paypal, Money Gram
Basic Info
- Model NO.: BAY175/1250-35U
- Filter Material: PE
- Type: Plate Filter Press
- Filter Area(M2): 50
- Filter Plate Number(PCS): 50
- Filter Plate Size(mm): 800*800*65
- Filter Cloth Material: Resistance to Acid and Base
- Support Beams: Q235 Steel Welding Parts (Thicker Rectangular Tube
- Transport Package: Wooden Case Package
- Origin: China
- Structure: Belt Type
- Pressure: Mechanical Pressure
- Namme: Filter Press
- The Thickness of The Filter Cake(mm): 30
- The Filter Room Number(PCS): 50
- Filter Plate Material: Reinforced PP with 1.0-2.5MPa Filtration Pressure
- Number of Filter Cloth(PCS): 51
- Trademark: LEABON
- Specification: 50M2
- HS Code: 8421291000
Product Description
Hot Selling ROTARY VACUUM DRUM FILTER PRESS
Detailed Product Description
Auto press and open by hydraulic
Auto feeding
Auto plate shifting
Auto bomb door and drip tray open and close
Automatic program-controlled chamber filter press has such features as follows
Technical Parameter
Application:As a intermittent pressure filtration equipment, it's used to separate the solid and liquid of suspension. Since the excellent effect of separation and convenient to use, it's widely used in petroleum, chemical, dyestuff, metallurgy, pharmacy, food, paper making, coal washing and sewage disposal and various industry which separation is needed.
PP filter press consists of high pressure plate, and filter chamber made of ordered filter plates. The slurry materials are sent to filter chamber by the pressure of feeding pump and then go into filtrating media (filter cloth) to separate solid and liquid in slurry material. When solid in slurry material becomes cake shape, The filter plates are molded by reinforced and thicken polypropylene, and they can bear on 1.0-2.0MPa filtration pressure.
Structure Features
Structure Features
Detailed Product Description
Auto press and open by hydraulic
Auto feeding
Auto plate shifting
Auto bomb door and drip tray open and close
Automatic program-controlled chamber filter press has such features as follows
Technical Parameter
|High pressure automatic hydraulic filter press
|Filter area(M2)
|The thickness of the filter cake(mm)
|filter chamber volume(L)
|Filter plate number(pcs)
|Reinforced pp
|The filter room number(pcs)
|The filter material and characteristics
|Plates Material: Reinforced and high pressure pp
Features: Acid, alkali, salt erosion, non-toxic, odorless, light weight, high strength, and effortless operation. Salt solution and polar solvents, good toughness, chemical stability, Plates using cone-shaped point Boss design, molding, with filtering speed, low dehydration cake, washed thoroughly uniform, fine processing of the sealing surface sealing performance intact.
|Filter plate size(mm)
|Filter plate material
|Reinforced pp with 1.0-2.5MPa filtration pressure
|Resistance to acid and alkali
|Filter cloth material
|Resistance to acid and base
|Number of filter cloth(pcs)
|Including sealing sheet
|Support beams
|Q235 steel welding parts (thicker rectangular tube)
|Keep plate
|Q235 solid steel plate welded pieces
|The thrust plate
|Q235 solid steel plate welded pieces
|Pressure head
|Q235 solid steel plate welded pieces
|Oil cylinder
|20 steel welding
|Hydraulic station
|Q235B steel welded
|Filter control cabinet
|Carbon paint
|Electrical control cabinet
|Intelligent control panel Relay control
|Filtration temperature
|Filter pressure(Mpa)
|Feeding way
|The thrust plate feeding in the middle
|Discharging belt conveyor
|Feeding pump
|Ceramic high-pressure piston pump
|Power: 11kw motor.
|Flow pressure: 1.0-2.5MPa.
|Drainage way
|Open flow
|According to customer's requirement
|Hydraulic station valve group
|Hydraulic valve group
|The oil pump of the hydraulic pressure station
|Plunger pump
|Oil pump seat
|Power station(kw)
|Oil cylinder pressure(Mpa)
|Oil cylinder maximum effective schedule(mm)
|The cylinder diameter(mm)
|Compact form
|Hydraulic automatic packing, manual unloading
|The filter cake was dropped by its own weight
|The whole machine size(mm)
|The paint coating standard
|All steel pretreated to remove the oxide layer rolling, rust and foreign matter, the inner surface coated with epoxy paint, do strength corrosion, the outer surface again primer, epoxy anti-corrosion coating twice.
|Manufacturing standards
|The performance parameters
|1. Function:carrying the filter plate, providing top tight pressure, under a certain pressure to ensure the seal and achieve filtration purposes
2, consists of: Pressing plate, thrust plate machine feet, and the feet of the cylinder block, cylinder system, the main beam (thick rectangular tube)
3. Rack material: Welded steel, 304 stainless steel rails
Filter plate
|1, function: Install the filter media to ensure normal filtration under pressure, can be fully realized backwash
2. Filter plate material: Reinforced Polypropylene
3. Handle material: Reinforced polypropylene and wear-resistant plastic
The hydraulic system
|1, function: to provide clamping force, clamping cylinder return and ensure the normal operation of the robot
2, consists of: tank, pump systems, manifold, hydraulic valves, tubing, pressure display and transmission systems
3. Maximum hydraulic pressure: 30MPa; rated pressure: 20MPa.
|Manufacturing Standards: JB / T4333-2005 "Van filter and frame filter technology"
Quality assurance: product safety in line with European CE certification and the German DIN standard.
Quality assurance system in line with ISO9001: 2008 standard
Environmental System certified to meet: ISO14001: 2004 standard


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Celebrating the spirit of burgeoning tech businesses, Butler Community College's Entrepreneurial Institute is hosting a networking breakfast from 7:30 to 9 a.m. on Thursday in Andover.Technology entrepreneur Ben Tyson, president and CEO of Time Trails, Inc. will share his story on how he built his future by focusing on the past. Tyson, who co-founded the Wichita-based web technology company with his father in the spring of 2009, created a new, state-of-the-art visualization method of imagining historical events based on time and geographical location around a virtual globe. The Entrepreneurial Institute at Butler is committed to the ideals and spirit of entrepreneurship. The Institute is a unique partnership bringing together the resources needed to assist entrepreneurs at any stage of their venture. It delivers innovative and relevant programming that supports the economic health of our region – where our students graduate and our stakeholders work.
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9/15/2017Less than 1 Month Until NACCE 2017! It's not too late to register...
9/4/2017Important Annual Conference Reminders
10/8/2017 » 10/11/2017NACCE2017 - Register Today!
10/25/2017October Member Webinar - Catalyzing Entrepreneurial Ecosystems in our Relationship Economy
10/27/2017HP LIFE Informational Webinar - October 2017
10/30/2017 » 12/22/2017October 2017- NACCE's Entrepreneurship Specialist Certificate Online Training
11/2/2017Fostering an Innovator Mindset in Students & Entrepreneurs Webinar {Presented by Innovator Mindset}
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