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Alternate splicing in exon 47 of the Purkinje cell calcium channel generates a splice variant with a five base pair insert (ggcag) before the stop codon in rat. This five base pair change the open reading frame of the exon 47 for resulting in an extended C-Terminal. Novel protein interaction at this region was hypothesised. Yeast Two Hybrid System was employed to screen against cDNA library to check for any protein interaction with 5 base pair insert region of exon 47. This project aimed to test the toxicity/ autoactivation of the baits in the yeast and to find the minimum concentration of 3-AT (3-amino-s-triole) at which it inhibits the HIS3 gene. The experimental result shows that there was no leaky expression of the HIS3 gene. The autoactivation/toxicity test results showed that the baits are less toxic than the control bait. The growth of non-interacting colonies in the Triple Drop Out media revealed that a more defined media should be used, demanding the repetition of experiment to obtain more convincing results.
1.1. Nervous System
The human nervous system consists of the Peripheral Nervous System (PNS) and the Central Nervous System (CNS). The PNS is formed of the cranial nerves and the spinal nerves. The central nervous system consists of the brain and the spinal cord. The brain can be divided into three major parts cerebrum, cerebellum and the brain stem. The cerebrum is divided into frontal lobe, parietal lobe, occipital lobe and the temporal lobe. The main function of cerebrum includes controlling of sensory organ, motor function, consciousness and imagining. The cerebellum is a uniform structure and its function is essential in movement and co- ordination of organs. The brain stem is made up of the mid brain, the pons and the medulla. The main functions of brain stem are transmission of information to and from the brain (Bear et al, 2001; Purves et al, 2004 and Thompson,1993).
1.2. Cells of CNS
The brain consist mainly two types of cells nerve cells or neuron cells and the glial cells. The neuron are involved in the transport of electrical signals from the brain whereas the glial cells are thought to be the supporting cells of neurons by the uptake excess of neurotransmitter that are essential for signalling between neurons (Henn et al, 1971 and Purves et al, 2004) and plays a role in synaptogenesis of the neuron (Bacci et al, 1999). The glial cells are of three types: astrocytes, oligodentrocytes and the microglial cells.
1.2.1. Glial Cells
Astrocytes are star shaped cells. The spatial arrangement of these cells between the capillaries and the neurons enables it in the modification of cellular responses, synaptic plasticity and survival of neurons (Abe et al, 2006 and Chen et al, 2003). Astrocytes important in glutamate transport, removal of free radical, controlling of haemostasis of brain and in maintaining a preferable environment for the active functioning of neurons by buffering K+ ions in their extracellular space (Chen et al, 2003; Gee et al, 2004 and Longuemare et al, 1999).
Oligodentrocytes are type of glial cells that insulate the neuron with myelin sheath (Bear et al, 2001 and Lubetzki et al, 1993. The myelin sheath is a membrane which is made up of lipid and two proteins the proteolipoprotein (PLP) and the myelin basic protein (MBP). (Colman et al, 1982 and Boison et al, 1995). At regular intervals myelin sheath becomes thinner and is known as Nodes of Ranvier (Peter et al, 1966). These regions are the site for voltage gated sodium channels and a number of proteins. Microglial cells are the macrophages of the brain, which are formed in the bone marrow and are then transported to the brain by specialized protein called chemokines (Khoury et al, 2008) The study of chemokine receptors is one of the important research areas in the pathogenesis of Human Immuno Deficiency Virus. HIV can target microglial cells for their replication (Albright et al, 1999; Ghorpade et al, 1997 and Meer et al, 2000). Microglial cells are also studied for their inflammatory responses in the brain. The identification of role and mechanism by which microglial cells cause inflammation has paved path for finding targets and therapeutics for many diseases.(Bhatia, 2008; Huang et al 2008; Hwang et al, 2008 and Kim et al, 2008).
Neuron or the nerve cells are units of the nervous system involved in transfer of electrical signal between each other and to the effector cells. There are many types of nerve cells. Purkinje cells are one among them (Brown, 1991). The study of calcium ion channel of Purkinje cell is the subject of this project.
The basic parts of neuron consist of a soma or cell body, axon, dendrites and neurites.
All neurons are covered by the neuronal membrane. The soma or the cell body is similar to any other type of cell in the body. The axon is fibre that transport signal from the cell body to other neuron or to the target cell. The axons are covered by myelin sheath of the glial cells. The axon may be branched or unbranched. The main function of axon is to transfer the electrical signal from the axon hillock of soma throughout the axon known as the action potential and to transfer the signals to other cell in the form of chemical signal, the neurotransmitter (Purves et al, 2004 and Bear et al, 2001). The region of contact with other cells where release of neurotransmitter takes place is known as the synapse. The release of neurotransmitter is facilitated by synaptic vesicles of the presynaptic terminal (one which release chemical signal). The neurotransmitters are released by the synaptic vesicle in the space between pre synaptic and post synaptic terminal known as the synaptic cleft (Purves et al, 2004 and Brown et al, 1991). The neurotransmitters are then received by specific receptors of the post synaptic terminal which would generate an action potential in the cell. Apart from these receptors the ion channels of the cell membrane of the synaptic terminal also respond in the transfer of signal. Dendrites are branched fibres that arise from the cell. Their surface is lined with number of receptor to receive signals for the neuron (Brown,1991., Purves et al, 2004., Thompson,1993 and Bear et al, 2001).
Purkinje cells are one of the largest types of neurons on the brain. They are found in the cerebellar region of the brain. The study of calcium ion channel of Purkinje cell is the subject of this project. Purkinje cells have a number of branches dendrites that receive synaptic inputs. As the dendrites receive signals it initiates a Ca2+ signal, which are important secondary messenger in the cells. The dendrites are the region for a calcium ion entry through the calcium ion channel. Similarly the soma contains K+ and Na+ channels(Schutter et al, 1994). These ions are of particular importance as their charge variation inside and outside the membrane trigger signalling in the cell. The transport of these ions is highly selective and they are maintained by the ion channel proteins of the Purkinje cell membrane and other neuronal membrane. These proteins form a pore for the transport of ions. Techniques such as the Patch clamp method have made the study of these ion channels easier (Bear et al, 2001).
1.3. Ion channel
Ion channels are glycoprotein complex that allow specific ions through them. The proteins of ion channel are coded by different gene. More than 100 genes are known to code ion channels. The transportation of ion is important in generating action potential in the cell and is also important as the ions are second messengers in signalling. Diseases associated with the ion channel are known as channelopathies. Ion channels can be three major types voltage gated ion channel. Ligand gated ion channel and the stretch and heat activated ion channel (Purves et al.,2004).
Voltage gated ion channels open and close on response to electrical potential. The voltage gated channels are made up of different protein sub unit. The subunits can move to open or close the channel (Horn, 2002). Depending on the type of ions they conduct they are further divided into Calcium channel, sodium channel and potassium channel. Ligand gated channels are those that respond to chemical signals. The ligand gated receptors are of five types nicotinic acetylcholine receptor (AChR), glutamate receptor, γ-aminobutyric acid (GABA), glycine-activated Channels and the ryanodine receptor(Stroud et al, 1990). Each of these receptors bind to specific ion and are found in different organs. The stretch and heat activated ion channel respond to heat or structural deformation of membrane (Purves et al, 2004).
1.4. Voltage Gated Calcium Channel (VGCC)
Ca2+ ions are important secondary messenger in cells and play important role in biochemical pathways of cell. The level and entry of these Ca2+ ions in the cell is highly regulated. The regulations of these ions are controlled by the Voltage Gated Calcium Channel (Gribkoff et al, 2006). These VGCC are mainly found in excitatory cells such as the muscle cells and neurons. They exert their function by controlling muscle contraction, neurotransmitter release, neuronal plasticity, synapses, and neuronal excitability (Pietrobon, 2005 and Yang et al, 2005) . VGCC respond to membrane depolarization facilitating Ca2+ entry into the cell and thereby activating the signalling cascade of the cell (Yang et al, 2005).
The normal functioning of the calcium channel protein is very important in a cell. Mutation in the gene coding channel protein, have been known to cause a number of diseases which include Timothy syndrome, Familial hemiplegic migraine type 2, episodic ataxia type 2, spinocerebellar ataxia type 6 and autism spectrum disorder which are grouped under “calcium channelopathies” (Bidaud et al, 2006 and Jen et al, 1999). Calcium channels also play a key role to mediate neuronal pain pathways (Gribkoff et al, 2006). A number of drugs have been known to block calcium channel and they are categorised as Calcium Channel Blockers. Verapamil was the first drug found to block Calcium Channel and later dihydropyridines (DHPs) class of drug was discovered to act as calcium channel blocker (Dolphin, 2006). DHPs are of much importance in studying the channel properties of the Dihydropyridine sensitive calcium channel. These DHP sensitive channels have dihydropyridine receptor for their binding (Campbell et al, 1988). Calcium Channel Blockers are now being found effective in the treatment of pain and hypertension (Atanassoff et al, 2000; Kize et al, 2001, and Thompson et al, 2001) but the question of safety in Coronary Heart Disease and the increased risk of cancer in patients remains unanswered (Eisenberg et al, 2004 and Fitzpatrick et al, 1997).
1.5. Calcium channel structure
A calcium channel consists of five important subunits α1. α2, β, δ and γ. The α1 subunit is known as the pore forming complex (Yang et al, 2006).
The α1 subunit is a single polypeptide and its functions mainly include voltage sensing, gating and selective permeation (Horn et al, 2000). The structure of α1 subunits consist of 24 segments (S1-S6) which constitute 4 domains, a C- terminal, N-terminal and Interlinkers. The linkers connecting domains are known as Loops and they are referred as loop I-II, loop II-III and loop III-IV depending on the domains they link (Dolphin, 2006).
The intracellular loop of the α1 subunit has interaction site for the binding β subunit. The interaction can modulate the G- protein, an important second messenger in the cell (Dolphin, 1998). The specific binding of β subunit to the tryptophan residue is important for controlling the gating of α1 subunit of certain type of channels (Berrou, 2002). S4 is another important segment of the calcium channel. It is the voltage sensitive region of the calcium channel. S4 segment moves outward causing the channel to open by getting depolarised. S4 segment is positively charged due to the presence of arginine aminoacid making it voltage sensitive by translocation of the charges across the membrane (Sigworthl, 2003 and Horn et al, 2000). The S5, S6 and the linker connecting the S5 and S6 segment forms the boundaries ion conducting pore of the α1 subunit. The ion conductance partly depends on the rotational movement of the S4 segment which either cause the S6 segment to open or close the pore (Horn et al, 2000).
The β subunits of the calcium channel are thought to be tissue specific and organ specific. Primarily they are of 4 different types, β1, β2, β3 and β4. Different isoforms of the β subunits also do exist which include (CaB2a, CaB2b and CaB3) (Hullin et al, 1992 and Petegem et al, 2006). Their association with α subunit is essential for modulation of VDI, CDI and CDF (Petegem et al, 2006). The α2 subunit is also known as the α2/δ subunit as both the subunits are product of a single gene (Petegem et al, 2006). The α2 and δ subunits are linked together by disulphide bonds. Like other subunits α2/δ also exists as isoforms (Wang et al, 1999). They are known to play an important role in plasticity of neuron after a nerve injury and neuropathic pain processing (Luo et al, 2001). Gabapentin is a drug known to act on α2/δ subunit, but their binding affinity varies with different isoforms of the δ subunit (Luo et al, 2001 and Luo et al, 2002).The γ subunit is found only in skeletal muscles. Their functional roles are yet to be discovered (Petegem et al, 2006).
The C-terminus of calcium channel is a site for a number of protein- protein interactions in some channels. The expansion of the polyglutamine tract of the calcium channel is a major reason for the pathogenesis of the disease, Spino Cerebellar Ataxia 6 (SCA6). The cell death in SCA6 is thought to be caused by the poisoning of the nucleus by the localisation of C-terminal fragments (Kordasiewicz, 2006).
1.6. Calcium Channel Types
Calcium channels account for the major amount of the calcium entry into the cell. The channel properties are tightly regulated to maintain Ca2+ concentration of the cell. The regulation was done through three well known processes.
Voltage Dependent Inactivation (VDI) – responsible for preventing entry of calcium into the cell. Calcium Dependent Inactivation (CD1) – responsible for preventing entry of calcium into the cell whereas Calcium Dependent Facilitation (CDF) – allows for the entry of calcium for signalling (Petegem et al, 2006).
Based on the amount of current required to activate the channel the VDCC were termed either LVA channel (Low Voltage Activated) or HVA channel (High Voltage Activated). Later on due to the discovery of different current types, location of channel and sensitiveness to different types VDCC were broadly classified. Thus now 6 different types VDCC are known, in T type the current is transient, located in T-tubules and sensitive to dihydropyridine (DHP) (Dolphin, 2006). In L-Type the current is long lasting, found in neuron, heart and skeletal muscles and are sensitive to DHP. The N-Type stands for Non L Type or Neuronal and they are sensitive to ω-conotoxin GVIA (Petegem et al, 2006). The current found in Purkinje cells of the cerebral cortex were P-Type, they were sensitive to ω -agatoxin IVA. The Q-Type current are found in granular cells, however scientist consider P-Type and Q-Type to be same and are now term as P/Q- Type. The difference between the P Type and Q-Type is thought to depend on the β subunit to which it is associated(Dolphin., 2006). Another type of Residual current was also discovered which to date is not sensitive to any of the known toxin, this current is known as R-Type (Dolphin, 2006 and Petegem et al, 2006).
1.7. Calcium Channel Gene
The alpha sub unit of the calcium channel are coded by 10 genes, therefore 10 different α1 sub units are known. Of the ten types Cav 1.1 – 1.4 which is found in L-type, Cav 2.1 or the Cavα1A is found in P/Q type channel, Cav2.2 is found in N type and Cav2.3 in R type channel. The Cav 3.1- 3.3 is found in T type channel. All these alpha subunit have one or more isoforms that would contribute to their functional diversity (Dolphin, 2006).
The gene coding for the Cav 2.1, CACNA1A is found on the chromosome 19p13. This gene belongs to CACN family of gene that code for calcium channel. The gene characterised by the extension of CAG trinucleotide repeats. In humans the extension of the may vary from 4 to 18. Mutation of this gene cause diseases cause three major diseases FHM1 (Familial Hemiplegic Migraine 1), EA2 (Episodic Ataxia 2) and SCA6 (Spino Cerebellar Ataxia 6).
Familial Hemiplegic Migraine is an autosomal dominant type of migraine caused by the missense mutation in CACNA1A. Three different mutations of CACNA1A cause FHM1 (Ducros et al, 1999). FHM1 affects the channel inactivation and the kinetics of the calcium channel (Kraus et al, 1997). The replacement of threonine with methionine is the mutation associated with FHM1. This mutation changes the channel structure causing more flow of calcium into cell. This ultimately results in the release of excess neurotransmitter (Ophoff et al, 1998). Episodic Ataxia 2 (EA2) is neurological disorder affecting the cerebellum and causing ataxia. The drug acetozolamide is known to be effective on EA2 (Ophoff et al, 1998). This disease has been found to have small but stable trinucleotide expansion but the role of the expansion is unknown for this disease (Jodice et al, 1997). The mutation in EA2 causes truncation of α1A subunit which might cause a complete loss of the function of the channel (Wappl et al, 2002).
1.8. Spino Cerebellar Ataxia 6
Spino Cerebellar Ataxia 6 is also a neurodegenerative disease caused by the increase in number of CAG repeats in the CACNA1A gene (Tanaka et al, 2000). The number of trinucleotide repeat is between 22 and 28 in SCA6 (Riess, 1997). But it is not only the CAG repeats that are causing the disease. The α1A have 6 isoforms and not all the isoforms are with the polyglutamine repeat. Therefore whether SCA6 is a channelopathy or Polyglutamine Disease remains a question among scientist (Frontali, 2006). The isoforms responsible for SCA6 is mainly limited to the C-Terminal. As the C-terminal is site for protein- protein interaction, changes in strength of interaction or changes in interacting partners tremendously affect the channel kinetics and other functional modification. As polyglutamine disease it cause toxic effect considered through aggregate formation (Pril et al, 2004). Comparison of number of repeats with other polyglutamine diseases where the repeat number is much high, the aggregate formation alone cannot account for pathogenesis (Matsuyama et al, 1999). As a channelopathy the degeneration of Purkinje cell is caused by the poisoning of nucleus with the localised fragments of C-Terminal. The cleaved C – terminal product is considered to have involved in signalling mechanism of the cell (Kordasiewcz et al, 2006). The isoforms of the C-Terminal of calcium channel are of considerable importance as the variation are found to be species specific (Kanumilli et al, 2005) and a few of them do not code for polyglutamine repeats. This invokes an interest in the C-terminal of the α1A subunit of the calcium channel. The isoforms are formed by a process known as the pre-mRNA alternate splicing.
Transcription of messenger RNA (mRNA) from DNA and translation of proteins from mRNA forms the central dogma of molecular biology (Crick, 1970). These processes involves a series of important events, one among them is pre mRNA splicing. Before translation of protein, the mRNA needs to be processed by removing of non-coding introns. A human gene on an average consists of 8 introns. Splicing can lead to more than one type of mRNA from a single gene and consequently different protein isoforms (Faustino et al, 2003). Many different proteins are involved in splicing most importantly the spliceosome, a complex formed of small nuclear RNA (snRNA) and small nuclear ribonucleoproteins (Hagiwara et al, 2005 and Jurica et al, 2003). Small nuclear RNA can be of 5 important types U1, U2, U4, U5 and U6. All these in different combination target specific pre mRNA. The targeting is based on a number of factors which include phosphorylation of snRNAs, catalytic metal ions, enhancers, transcriptional coregulators and serine/arginine rich SR protein (Shi et al, 2006; Saba et al 2005; Auboeuf et al, 2007; Jurica et al, 2003; Hicks et al, 2005 and Manley et al, 2006).
In general a spliceable introns has three regions splice donor, splice acceptor and a branch site. Most of the splice donor regions consist of AU nucleotide and the splice acceptor region consist of AG (Kenneth., 2005). Spliceosomes attach to these ends and by transesterification remove the introns, followed by the ligation of the exon (Rio,1993). Several mRNA have inherent splicing mechanism that does not require any spliceosome as they can splice themselves known as self splicing (Herrin et al, 1990 and Landthaler et al, 1999). Though most of the splicing is limited within the same mRNA, splicing also occurs between two different mRNAs by trans-splicing mechanism. The two mRNA exons called the mini exons were transcribed in different gene and were then combined to translate for a single protein (Bonen, 1993 and Bonen, 2008).
Alternative splicing is a mechanism by which a few genes produce innumerable proteins that are diversified in structure and function. Nearly 75% of the human genes are involved in alternate splicing to give different protein isoforms (Hagiwara et al, 2005 and Stamm et al, 2004). The needs to understand alternate splicing have arised in almost all fields of biology. In evolutionary terms alternate splicing has a major role in the functional development of species right from the times of “RNA world”. The importance of isoforms has been understood through a number of studies. The Active and inactive forms of Sex lethal protein isoform are the determinants of sex of Drosophila (Herbert et al, 1999; Irimia et al, 2007 and Poole et al, 1998). Many different isoforms of normal proteins are discovered in cancer cells. These studies of these isoforms and their role have revealed some important diagnostic approach and cancer cell biomarkers (Brinkman, 2004; Skotheim et al, 2007 and Pampalakis et al, 2008). In the drug discovery process it is necessary to consider the mechanism of protein isoforms and pre mRNA splicing pathways and signalling molecules to identify new targets for drugs (Levanon et al, 2003 and Hagiwara et al, 2005).
Alternative splicing in ion channels alter the conductance and functional properties of the channel. Splicing has been known in voltage gated sodium channel, voltage gated calcium channel, ligand gated ion channel and in calcium gated potassium channel. Although the ion channels differ in their properties, all share some basic function. These ion channels have multiple splicing site through which their channelling properties are regulated based on the organs where these channels are located (Copley, 2004; Raymond et al, 2004; Sarao et al, 1991 and Schaller et al, 1992).
1.10. CaV2.1 splice variants
Variants in calcium channel protein, in particular the 47 exon of the c-terminal is the basis of this study. Splicing in calcium channel occurs at distinct region such as the loops between the II-III domains which is the major interacting site for ryodine receptor. Two isoforms BI and rbA are found in loop II-III of rat and rabbit. They differ in their interacting ability towards syntaxin and synaptotagmin proteins. These proteins can modulate the Ca+ influx of the neuron (Charvin et al, 1997 and Rettig et al, 1996). Site specific variations are found in exons 9, 31, 44, 46 and the extreme C terminus e47 (shown in Fig 3)(Kanumilli et al, 2005).
The C- termini of calcium channels are involved in the modulation of G-proteins, molecular switching of calmodulin and are the site for protein-protein interaction. So a single amino acid change can potentially change the gating property and other function of the channel and its interacting partners (Chaudhuri et al, 2004; Gray et al, 2007; Krovetz et al, 2000 and Ligon et al, 1997) splice variants were known to occur in the C- terminal the calcium channel. A 5 base pair insertion (ggcag) was reported in pancreatic islets of rats a variant already known in human (Ligon et al, 1997). This 5 base pair insertion is expected to alter the length on the c terminal and hence channel property as it found before the stop codon, which means a change in the reading frame. The existence of variants with and without the 5 base pair (ggcag) insert before the stop codon of rat Purkinje cell is confirmed by Kanumilli et al (2005).
Another independent study with mouse by Tsunemi et al (2001) also confirmed the 5 base pair insert. In addition, variants without the stop codon and a ggcag insert, 150 nucleotide deletions in the 5′- end of the C- terminal is reported in mouse (Kanumilli et al, 2005). The absence of stop codon was also observed in the study by Tsunemi et al (2001) in mouse. Richards et al (2007) obtained similar results with rat Purkinje cell, the sequence of exon 47 were same as the rat pancreatic cells except for variations in other exons. However variation in the number of amino acid (156 residues, 153 residues and 115 residues) coded by exon 47 were observed in different clones. The 156 amino acid length was also reported by Ligon et al (1998).
These finding and most other results describe the calcium channel properties in terms of activation or inactivation kinetics. However no protein- protein interaction study is available till date for the exon 47 with five base pair (ggcag) inclusion before the stop codon. The need for studies at the protein-protein interaction level is necessary which is evident from the studies of Dolphin(2006), Richards et al (2007), Sandoz et al (2001) and Kanumilli et al (2005). This study was aimed at studying possible protein-protein interaction for exon 47 of rat Purkinje cell. Then linking the interacting the protein to already known biochemical pathway is expected to give more insight the channel and possibly a new perspective in the treatment of SCA6.
1.11. Protein – protein interaction studies
Protein-Protein interaction is an important part in all biological process. A protein- protein interaction can altogether change the binding characteristics, kinetic property and their catalytic ability (Eisenberg et al, 2000). A number of methods have been developed and used to study protein-protein interaction. These methods can be the detection and analysis of interaction or can be screening against a family of proteins. Detection methods are mostly used to confirm and study known interaction. These methods include Protein Affinity chromatography, Affinity Blotting, Coimmunoprecipitation and Cross- linking. The screening methods include protein probing, phage display and the Yeast Two Hybrid System (Y2H) (Phizicky et al, 1995). Bioinformatics tools such as protein docking are also important in predicting the protein interactions (Smith et al, 2002).
1.12. Yeast Two Hybrid System (Y2H)
Yeast two hybrid system is the most widely used protein screening methods. The requirement of an interaction between two domains DNA Binding Domain (DNA-BD) and Activation Domain (AD) for the expression of a reporter gene (lac-z) in yeast is being exploited in Y2H. The lac-z gene expression gives our β-galactosidase enzyme which can be observed by colour change confirming interaction (as shown in Fig 4) (Criekinge et al, 1999).
The protein of interest (bait) is usually fused with the BD and the interacting protein or the library protein is fused with activation domain. The protein of interest is normally termed as bait and the interacting protein is called a prey. Bacterial plasmid can be easily constructed to express fusion protein of interest. The bacterial shuttle vector can be isolated and transfected into the yeast for their expression. On expression the DNA-BD fusion protein will bind to the upstream activation sequence of the reporter gene. Two types of Y2H are known one is the GAL4 based system and the other is the Lex A based system. In Lex yeast two hybrid system the prey is fused with the Lex A binding domain. The specifically interacts with the Lex A operator upstream sequence which is the part of the promoter for reporter gene. The prey will be fused with the GAL 4 protein. In the GAL 4 system instead of Lex A the GAL 4 promoter will be used. Both the systems have their advantages and their disadvantages (Criekinge et al, 1999 and Luban et al, 1995)
The yeast strain L40 is compatible with LexA operator and the GAL 4 promoter system. Most Y2H methods are done more than one reporter gene for more selectivity. HIS3 gene is one such reporter that is used for the nutritional selection of the cells. HIS3 reporter expression needs the interaction of proteins. So cells would not grow in a media lacking histidine if no interactions take place. Similar nutritional selections are also used in cell containing only the baits or only the prey. The nutritional selection for bait is tryptophan and for the prey is leucine. It is therefore important to use a defined media. A positive interaction between bait and the prey will allow growth in the Triple Drop Out media (TDO/ -His/-Leu/-Trp) (Criekinge et al, 1999 and Luban et al, 1995)
The use of histidine reporter gene can sometimes account for leaky expression. In which case 3-AT (3-amino-s-triole) a competitive inhibitor of histidine can be tried in various concentration to find a minimum concentration at which cells grow and the enzyme is inhibited. Cells growing concentration of 100mM concentration cannot be used as baits (Criekinge et al, 1999).
Toxicity caused by bait can inhibit the growth of yeast (Zhong et al, 2003). Toxicity tests have to be carried out to after the baits are designed. Autoactivation of the baits should be checked before proceeding to the, library screening as nearly 5% of the protein can initiate transcription without an interactor (Criekinge et al, 1999). After the library screening the plasmids can be isolated and used to transform bacterial cells. The interaction also has to be confirmed and isolated by techniques such as coimmunoprecipitation.
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Amazon Prime members are getting another perk, and it's something mobile gamers would appreciate. The e-commerce giant is making free mobile game loot a part of its subscriber offerings, starting with exclusive in-game items for Tencent's popular battle royale game PUBG Mobile. It has already dropped the Infiltrator Mask, which will be available until October 3rd, and will release more items one by one in the coming weeks. The other items in the Prime exclusive set are the Infiltrator Jacket, Shoes and Pants, as well as the Epic Level Gun and Magma Parachute.
Amazon told TechCrunch that it will roll out more mobile gaming content for different titles in an ongoing basis. It already has a list of future partners, including EA, Moonton, Netmarble and Wargaming Mobile, so subscribers can expect to get goodies for those companies' mobile titles in the future.
Prime members already have access to gaming-related perks, such as free games and a year of Switch Online, via Twitch Prime. It focuses more on PC and console gaming, though, which is why Ethan Evans, the VP for Twitch Prime, said in a statement: "Now, no matter what platform you play on -- whether console, PC, or mobile -- there are Prime game benefits for you." He added: "We're starting with exclusive content for PUBG Mobile, one of the biggest mobile games in the world, and in the coming months, we'll roll out benefits for some of the most popular mobile games across many favorite genres."
PUBG Mobile players with Prime subscriptions can claim their free items by going to Amazon's official page for the campaign using an Android or an iOS device.


Here’s a warning to the Senate minority leader, Chuck Schumer, and other environmentally inclined Senate Democrats, like Sheldon Whitehouse and Ed Markey: A legislative minefield lies dead ahead, pocked with destructive amendments of Republican origin hostile to clean air, clean water, endangered species and fragile landscapes. And here’s a plea: Stop these measures from becoming law.
Following its approval of the big budget deal on Feb. 9, Congress began writing the dozen appropriations bills that direct federal dollars to specific agencies. These bills are likely to be incorporated in one giant omnibus spending measure to be negotiated over the next few weeks by House and Senate leaders in advance of the March 23 expiration of the continuing budget resolution that has kept the government going.
Given its urgency, the bill is fertile ground for the kind of mischief the Republicans in particular have been notorious for over the years—loading up must-pass bills like this one with provisions, known as riders, that in most cases could not survive on their own and thus need protective cover. In years past, such riders were usually inserted at the last minute on the House or Senate floor. Here they are in plain sight, having been approved in earlier votes or endorsed by powerful committee chairmen or chairwomen who will do their level best to make sure they are included in the final bill. Mr. Schumer can prevent that from happening. The Democrats are effectively 49 in number, the Republicans 51. By holding his party together, he can deny the Republicans the 60 votes they need to overcome a filibuster—ensuring a clean bill, and a cleaner environment.
Public interest groups have counted nearly 90 of these riders, but here are several of the worst:
Clean water: In 2015, the Obama administration adopted a landmark rule intended to clarify and broaden protections for smaller streams and wetlands vital to the country’s drinking water and wildlife. Developers and big farmers complained to Scott Pruitt, the industry-friendly head of the Environmental Protection Agency, who has begun the lengthy process of replacing the rule with something more favorable to commercial interests. That’s not fast enough for the leaders of three separate appropriations panels pushing nearly identical riders that would kill the rule right away, without consulting the public or conducting the scientific analysis required by law.
Methane emissions: As part of its larger strategy to combat climate change, the Obama administration approved two rules to minimize emissions of methane, a potent greenhouse gas. The EPA would regulate emissions from new oil and gas wells; the Interior Department would require oil and gas companies to control venting and flaring from existing wells on public lands. Efforts to delay (and ultimately rewrite) both rules have been thwarted by the courts. Here again, Congress comes to the rescue with two riders (both approved in earlier House floor votes) that would kill both rules.
Alaska wild lands: Lisa Murkowski, Republican of Alaska and chairwoman of the Senate interior and environment appropriations subcommittee, managed to sneak a hugely controversial amendment into last year’s big tax bill opening up the Arctic National Wildlife Refuge for oil drilling. Her amendment would never have passed as a stand-alone measure. Now she wants more. One amendment she seeks would weaken protections against the clear-cutting of old growth trees in the Tongass National Forest. Another would exempt forests throughout Alaska from one of the most significant forest conservation measures of the last century, the Clinton-era “roadless rule” forbidding road building and, by extension, logging, mining and other commercial activity on roughly 50 million acres of wild national forests.
—The New York Times


FORT JACKSON, S.C. – In 2008, Charleston District reassumed responsibility for military construction projects at Fort Jackson. Since then, the district has executed over $1.4 billion in projects including new builds, operations and maintenance, and facility investments services.
Many of those projects were aimed at improving the basic training environment. Fort Jackson is the largest of the four basic combat training installations in the Army and roughly 60% of all new Soldiers are trained there.
Command Sgt. Maj. Chad Blansett, the U.S. Army Corps of Engineers South Atlantic Division command sergeant major, recently toured the various work sites. Blansett was joined by Command Sgt. Maj. Philson Tavernier, command sergeant major for Fort Jackson, Command Sgt. Maj. Roberto Guadarrama, command sergeant major for the 165th Infantry Brigade, and Command Sgt. Maj. Travis Wirth, the command sergeant major for the 193rd Infantry Brigade.
“One of the most important parts of my job is to make sure the senior enlisted advisors in our Army have a better opportunity to understand what USACE does for them, especially in our region with all the military construction projects,” said Blansett. “As I travel around the southeast, I’m able to tie in the things we are doing for our Army to those who are training America’s best.”
The 165th and 193rd are the two brigades that conduct basic combat training at Fort Jackson.
“I appreciate the opportunity to have had the occasion with the project walk-through,” said Guadarrama. “It was extremely educational. There has been an extraordinary amount of effort from team members and team leaders from different organizations in making these projects come to life.”
One of the projects currently underway is Basic Training Complex Four, Phase Two, which was started this past June and is scheduled to be completed by the end of this year. Formally known as a Basic Combat Trainee Complex, it has all the required facilities to house and fully train the Army’s new recruits, while also being close to field training areas and ranges.
“Fort Jackson is the first point of contact for a large percentage of the Soldiers who enlist in the U.S. Army and it’s very important we provide the optimal onboarding to the newest members of our team,” said Tavernier. “These new facilities will allow us to consolidate battalions into one organic location with the ability to train at scale without any impeding to our mission while enhancing capabilities.”
Fort Jackson graduates an average of 1,000 basic combat training Soldiers each week and on any given day, roughly 10,000 people are training on Fort Jackson.
Following the walk-through, Blansett took some time to meet with staff at the Corps’ Fort Jackson Resident Office.
“Words cannot express how much I appreciate the team and the work they do,” said Blansett. “A lot of great work goes on behind the scenes and it doesn’t go unnoticed. I certainly appreciate what everyone does to make USACE as great as it is.”


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Distinguished Department of Forest Resources professor Peter Reich was quoted in Great Lakes Echo on findings from the recent study Even modest climate change may lead to major transitions in boreal forests, which he was lead author of.
The purpose of this project was to inform unit leadership and visitor use management at Pictured Rocks National Lakeshore (PIRO) regarding boat behaviors and density at areas between Miners and Chapel Beaches.
This study compared the relative importance of the transport of raw forest products by trucks to the top five non-timber commodities and the fatality rates of log trucks to other heavy trucks in the lower 48 states, compared available national road damage cost estimates for interstate and non-interstate roads, and assessed the impact of relaxing interstate weight limits on hauling distance, travel time, safety, pavement damage, and CO2 emissions for hauling timber along three travel corridors in Wisconsin and Minnesota.
From Minnesota to the Northwest Territories, researchers are studying dramatic changes in the vast northern forests: thawing permafrost, drowned trees, methane releases, increased wildfires, and the slow transformation of these forests from carbon sinks to carbon emitters. Forest Resources Professor Lee Frelich, PhD, talks more about this important work.
The Fall 2022 edition of the Forest Scene newsletter is out, featuring stories of the department's students, alumni, and faculty.
From the Fall 2022 edition of the Forest Scene newsletter, an update on the Plant for the Future project, began two years ago at Crosby Farm Regional Park, St. Paul.
Theoren Keenan, a senior majoring in Forest and Natural Resource Management (B.S.), talks about their path to studying forestry here at the University of Minnesota.
From the Fall 2022 edition of the Forest Scene newsletter, Dr. Diana L. Karwan discusses her research studying water movement and watershed function.
From the Fall 2022 edition of the Forest Scene newsletter: Alumni and donor Dr. William A. Patterson III shares his deep commitment to teaching about fire ecology.
From the Fall 2022 edition of the Forest Scene newsletter: A letter from Dr. Kristen Nelson, Professor and Interim Head of the Department of Forest Resources, to our community.

domain: geography


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- 1 can (32 ounce-48 ounce) canned crushed tomatoes
- 1 jar (16 ounce) favorite tomato sauce
- 1 tbl. sugar
- 1 tsp dried basil
- 1 tsp kosher salt
- pinch red pepper flakes
- 1 tsp Mrs. Dash - original
Heat all the ingredients and season to taste with additional salt or pepper.
(may dilute with water if it simmers too long)
Add: 2 cups heavy cream
whisk until well blended
Top with shredded parmesan cheese and freshly chopped parsley.
May be served with garlic croutons.

domain: food


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DEREK J. KING, M.B.; JOHN G. KELTON, M.D.
Heparin-associated thrombocytopenia is a relatively common complication of heparin therapy occurring in approximately 5% of the patients who receive this drug. The incidence is higher with bovine heparin than with porcine heparin. Onset of heparin-associated thrombocytopenia usually occurs 6 to 12 days after initiation of treatment and by itself has a low morbidity. Heparin-associated thrombocytopenia plus arterial thrombosis can cause major complications including stroke, heart attack, and death. The incidence of heparin-associated thrombocytopenia plus arterial thrombosis is lower than that for heparin-associated thrombocytopenia alone. The diagnosis of heparin-associated thrombocytopenia remains one of exclusion, but testing for the presence of a heparin-dependent platelet-aggregating factor may prove to be useful. Analysis of the time of onset suggests a strategy for prevention. Oral anticoagulants could be started concomitantly with the heparin so that it could be discontinued in several days. This approach may prevent most episodes of heparin-associated thrombocytopenia.
KING DJ, KELTON JG. Heparin-Associated Thrombocytopenia. Ann Intern Med. 1984;100:535–540. doi: https://doi.org/10.7326/0003-4819-100-4-535
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Published: Ann Intern Med. 1984;100(4):535-540.
Coagulopathies, Hematology/Oncology, Platelet Disorders.
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Print ISSN: 0003-4819 | Online ISSN: 1539-3704
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domain: literature


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U.S. Treasury yields fell to their lowest levels since November on Monday after a report showed that U.S. manufacturing grew at a substantially slower pace in January.
New order growth plunged by the most in 33 years, driving overall factory activity to an eight-month low. The report sent Treasury prices back into positive territory, after the bonds had earlier weakened on solid European growth and some calming in emerging market assets.
"Overall this is a weak number and it does suggest some dramatic slowing in economic activity," said Millan Mulraine, deputy head of U.S. research and strategy at TD Securities in New York.
"One can put most of this down to the unseasonably cold weather that we've had over the past two months, we've seen that in other economic reports. The big question is whether that proves to be temporary and we think that it will," he said.
Treasury yields had earlier edged up after data showed that manufacturers around the world enjoyed a solid start to the year as order books swelled, though a struggle for growth in China and a downturn in France took the shine off the overall picture.

domain: news


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Australia’s Oceanlinx, whose home-grown, commercial-scale wave energy converter technology was unveiled with some fanfare last October, has been placed in receivership after the Sydney-based company hit troubled waters in February.
Rahul Goyal, one of two receivers appointed to the case from KordaMentha, said the company had “suffered financially” after an incident at sea several weeks ago delayed the final installation of its 1MW GreenWave wave energy converter – billed, at the time, as the world’s first such machine to be deployed.
The commercial-scale unit was damaged en route to its destination of Port MacDonnell, in the south-east of South Australia. This caused delays in funding, said Goyal, which was dependent on meeting installation deadlines.
Oceanlinx’s plan had been to install the 24m by 21m, 3,000 tonne unit 3km offshore and transfer the electricity it generated to the grid via a subsea cable. Once operational, the 1MW turbine was expected to produce enough electricity to power 1000 homes.
Instead, the commercial-scale unit, which sits on a base of prefabricated reinforced concrete, was towed into shallow waters at Carrickalinga, where it remains.
Formed more than 15 years ago, Oceanlinx was a promising player in Australia’s ocean energy sector, having a number of wave power prototypes, including three units off the NSW coast, and had plans to expand to North America, Asia and Europe.
The company’s award-winning “greenwave” technology – which works by using waves to produce high pressure air, which is converted into electricity by a turbine – was named in a 2012 CSIRO study as one of Australia’s most promising and advanced home-grown wave energy technologies.
The product of an $8 million project, it received $4.4 million in funding from the Australian Renewable Energy Agency under the Emerging Renewables Program.
In a media statement issued by KordaMentha Restructuring on Wednesday, Goyal said it was too early to outline the receivership strategy for Oceanlinx, but that the immediate priority was to work with state governments to alleviate any safety concerns. The company’s six employees would remain at work for now, he said.


Strain Gauges Hooke’s Law Example: Bone has a Young’s modulus E of about 100,000 psi. 10 psi of stress will stretch a 20 inch femur Compare to F = K*Δx …
stress-strain curve:stiffness, yield strength, fracture www.tpub.com/doematerialsci/materialscience22.htm
Transverse Strain and Poisson’s ratio Consider incompressible material (constant volume…)
What to expect on the strain gauge quiz: Given stress, Young’s modulus, gain, Vex, Poisson’s ratio, calculate Vout… www.solware.co.uk/blog
Another deformation: torsional twist http://home.gna.org/getfem/shots.html
What about the piezoresistivity term?It is normally not zero. In fact for some semiconductors it dominates the expression on the right of the equation. For metals, with a Poisson ratio around 0.3, nonzero piezoresistivity of 0.4*ε basically results in approx: In the Strain Gage Quiz the gauge factor will be 2.
from www.sensorland.com from www.natmus.dk
Unknown material properties vs strain gauge material properties: Gauge factor: of the strain gauge Poisson’s ratio: of the unknownstrain: of the unknownYoung’s modulus = Ym… Young’s modulus: of the unknown > Ym of SG Next consider a Wheatstone bridgemade with 4 strain gauges… Two voltage dividers to consider:
That is the term to finish the expression for Vout = G*Vex*ΔR/4R for one longitudinal strain gauge in a "positive" location in the bridge.
Sign of influence on Vout given SG placement: Table reads: if R1 is the LG strain gauge then how does Vout change? *dR/4R …
SG web links: • http://me.queensu.ca/courses/MECH215/documents/MECH215-Week06Lecture1-StrainMeasurement-Introduction.pdf • http://www.eidactics.com/Downloads/Refs-Methods/NI_Strain_Gauge_tutorial.pdf • http://www.thorlabs.com/images/TabImages/Strain%20Tutorial.pdf
FINAL REPORT TO THE NATIONAL OPERATING COMMITTEE ON STANDARDS FOR ATHLETIC EQUIPMENT (NOCSAE) 1999 “THE USE OF SHOE ORTHOTICS TO REDUCE BONE STRAIN RATE” http://www.nocsae.org/research/rep/Burr_final.htm


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Since the ratification of the Convention Сoncerning the Protection of the World Cultural and Natural Heritage by Verkhovna Rada of Ukraine in 1988, Ukraine has been working closely with the UNESCO World Heritage Centre. The World Heritage List includes seven Ukrainian cultural and natural properties:
"Kyiv: Saint-Sophia Cathedral and Related Monastic Buildings, Kyiv-Pechersk Lavra" (inscribed in 1990);
"L'viv – the Ensemble of the Historic Centre" (1998);
The transboundary site of scientific and cultural heritage "Struve Geodetic Arc" (geodetic points Baranovka, Katerinovka, Staronekrasovka, Felshtin, 2007) covers 10 countries: Belarus, Estonia, Finland, Latvia, Lithuania, Norway, Republic of Moldova, Russian Federation, Sweden and Ukraine;
"Residence of Bukovinian and Dalmatian Metropolitans" (now the building of the Yuri Fedkovich Chernovetsky National University, 2011);
"Wooden Tserkvas of the Carpathian Region in Poland and Ukraine" (Ukraine-Poland nomination, 2013);
"Ancient City of Tauric Chersonese and its Chora" (2013);
"Ancient and Primeval Beech Forests of the Carpathians and Other Regions of Europe" (2007).
The natural site "Primeval Beech Forests of the Carpathians"(Ukraine and Slovakia) was inscribed on the UNESCO World Heritage List as a transnational serial natural property of ten separate components and as an outstanding example of undisturbed, complex temperate forests exhibiting the most complete ecological patterns and processes of pure stands of European beech. In 2011, during the 35th session of the World Heritage Committee, the property was extended through addition of the Ancient Beech Forests of Germany. The tri-national property was renamed as the "Primeval Beech Forests of the Carpathians and the Ancient Beech Forests of Germany" (Slovakia, Ukraine, Germany). In July 2017, the transboundary extension of the World Heritage site of the "Primeval Beech Forests of the Carpathians and the Ancient Beech Forests of Germany" stretched over 12 countries (Albania, Austria, Belgium, Bulgaria, Croatia, Germany, Italy, Romania, Slovakia, Slovenia, Spain, Ukraine). It is now known as "Ancient and Primeval Beech Forests of the Carpathians and Other Regions of Europe". In 2020, Bosnia and Herzegovina, Czech Republic, France, Italy, Montenegro, North Macedonia, Serbia, Slovakia and Switzerland submitted a dossier for extension of this site to the UNESCO World Heritage Centre. The issue will be considered at the 44th session of the UNESCO World Heritage Committee in 2021.
A Tentative List is an inventory of those properties which each State Party intends to consider for nomination.
States Parties are encouraged to submit their Tentative Lists, properties which they consider to be cultural and/or natural heritage of outstanding universal value and therefore suitable for inscription on the World Heritage List. Nominations to the World Heritage List will not be considered unless the nominated property has already been included on the State Party's Tentative List and are removed from the Tentative List when they are inscribed on the World Heritage List of UNESCO.
As of 2021, the Ukrainian government has proposed 17 properties for inclusion on the UNESCO World Heritage List.
Cultural heritage sites:
Historic Centre of Tchernigov, 9th -13th centuries (1989)
Cultural Landscape of Canyon in Kamenets-Podilsk (1989)
Bagçesaray Palace of the Crimean Khans (2003)
Archaeological Site "Stone Tomb" (2006)
Mykolayiv Astronomical Observatory (2007)
Complex of the Sudak Fortress Monuments of the 6th - 16th centuries (2007)
Astronomical Observatories of Ukraine (2008)
Historic Center of the Port City of Odessa (2009)
Kyiv: Saint Sophia Cathedral with Related Monastic Buildings, St. Cyril's and St. Andrew's Churches, Kyiv-Pechersk Lavra (extension of Kyiv: Saint-Sophia Cathedral and Related Monastic Buildings, Kyiv-Pechersk Lavra) (2009)
Trading Posts and Fortifications on Genoese Trade Routes. From the Mediterranean to the Black Sea (2010)
Cultural Landscape of “Cave Towns” of the Crimean Gothia (2012)
The historical surroundings of Crimean Khans’ capital in Bakhchysarai (2012)
Tyras - Bilhorod (Akkerman), on the way from the Black Sea to the Baltic Sea (2019)
Mixed cultural, natural, and industrial heritage sites:
Tarass Shevtchenko Tomb and State Historical and Natural Museum - Reserve (1989)
Dendrological Park "Sofijivka" (2000)
Derzhprom (the State Industry Building) (2017)
Natural heritage Site:
National Steppe Biosphere Reserve "Askaniya Nowa" (1989)
Find out more information on the UNESCO web site: https://whc.unesco.org/en/statesparties/ua.

domain: history


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What Does a Psychiatrist do?
A psychiatrist is a mental health professional who can diagnose and treat mental health conditions. These professionals are medical doctors (M.D.) who specialize in understanding psychiatric conditions including substance use disorders, though there are many reasons why someone might see a psychiatrist, including conditions like depression, anxiety, bipolar disorder, personality disorders, and others.
All psychiatrists have an undergraduate degree, usually in psychology or pre-med topics. From there, they attend medical school for four years until they receive their medical degree. After that, like all medical doctors, psychiatrists must complete a residency program where they work closely under the supervision of more experienced doctors. Psychiatrists may also choose subspecialty practice areas, like drug treatment, school psychiatry, or veteran’s care. They must also hold all the proper licenses required by the state and will need to renew this license regularly and earn continuing education credits to keep their credentials up to date.
Psychiatrists can choose to work for a hospital or clinic or open their own private practice. Some psychiatrists even work in settings like prisons, military facilities, schools, rehabilitation centers, or other healthcare facilities. Psychiatrists are unique in the mental health field because they are able to diagnose mental health conditions and provide prescription treatment like antidepressants or anti-anxiety medication. Because psychiatrists are fully trained in medicine, visiting with a psychiatrist can also help you rule out any other medical issues that may be causing your mental health problem.
Psychiatrist Near Me
Seeking help for any mental health condition can be scary, especially if you’re facing it alone and don’t know where to start. You may fear that the people you love and care about may judge you negatively if they find out you’re seeking psychiatric care. Often, these patients want their treatment to be discreet, and seeing a psychiatrist nearby can help. If you’re wondering “”Where can I find a psychiatrist near me?”” our directory can help you find the treatment that’s right for you.
We have listings from hundreds of psychiatrists across the country to make it easier to search for one who is local to your area. Using our directory, you can find the location of psychiatrists and more information about their practice. When you seek treatment from a psychiatrist locally, you won’t have to travel as far to get the help and support you need. This means you can spend less time traveling to get treatment and more time with friends and loved ones while you stay focused on getting better.
If you are experiencing issues with drug or alcohol use or addiction, a psychiatrist can make sure you receive the proper care. Here are some of the greatest benefits of seeing a psychiatrist when you’re struggling with addiction:
● They can get you a diagnosis – oftentimes, substance abuse and mental illness co-occur. Those with mental illness may use substances to cope with their symptoms and find relief. A psychiatrist can help to diagnose underlying mental health conditions that may be at the root of your substance abuse.
● They can help you detox – detoxing is the initial stage in your recovery from addiction, and is often the most difficult part to get through. Drugs affect your brain chemistry, and a psychiatrist is an expert in just that. They can help you get through this tough stage without risking relapse.
● They can prescribe medications – sometimes, depression, anxiety, bipolar disorder, PTSD, or other mental illnesses are at the root of your drug abuse issues. A psychiatrist can help you find a medication that can help relieve some of your symptoms so you don’t have to turn to drugs or alcohol for relief.
When you see a psychiatrist for drug abuse treatment, they can help you understand exactly why you started using drugs and can help you find healthier ways to cope with stressors in your life.
Difference Between Psychologist and Psychiatrist
When looking for mental health professionals to help treat your substance abuse, you will likely have the option of visiting a psychologist or a psychiatrist. It’s important to understand the difference between these two professionals. As discussed above, a psychiatrist has received their medical degree and has completed an undergraduate program, a medical school program, and a residency.
A psychiatrist, on the other hand, has not attended medical school and has not gone through a residency program. They usually receive undergraduate degrees in psychology, and then a master’s degree or Ph.D. in psychology. Like psychiatrists, psychologists must be properly licensed by the state to practice legally, but unlike psychiatrists, they cannot prescribe medication.
Psychiatrists specialize in diagnosing mental illnesses and helping their patients manage treatment. On the other hand, psychologists tend to focus on providing their patients with specialized counseling or talk therapy, which provides their patients with a safe space to talk about their issues. Psychologists can also help their patients learn healthy ways to cope with problems in their lives and change potentially harmful patterns in their thinking and behavior. Sometimes, patients see both psychiatrists and psychologists in conjunction, so they can receive prescription medicine and therapy as part of a comprehensive treatment program.
Psychiatrist vs Therapist
In addition to psychologists and psychiatrists, you may also have the option to see a therapist. Therapists are mental health professionals who can help their clients overcome difficulties they may be facing in their lives. A therapist may specialize in family therapy, marriage counseling, addiction counseling, or other specialties. Sometimes, a therapist may have a master’s degree or doctorate in social work, counseling, or other fields.
In general, a therapist is more of an umbrella term that covers different kinds of professional counseling. Therapists can use techniques like cognitive behavioral therapy (CBT), dialectical behavioral therapy (DBT), or talk therapy to help their clients find relief from their problems. Finding a mental health professional who specializes in helping their patients overcome drug abuse and addiction can be the first step on your path to recovery. These professionals have helped many people like you overcome their drug issues and live a healthy and happy life.

domain: medical


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- BiographyOver his 1 year pornographic career, this latin male model has starred in 1 video, debuting in Black On White 4 around 2013. Content starring this star was requested 8,662 times in total, with more than 0.1 Gigabytes of porno having been downloaded during the previous year. Some other actors co-starring together with Joey include, most notably, Sophie Dee, Japan, Fernanda Alencar, and Brooke . His most recent scene is included in Sex Driven Girls. His favorite actions include Blowjobs, Handjobs, Anal Sex, Fingering, and Hardcore.
- Favorted0 times

domain: entertainment


text:
A weekend getaway takes an unexpected turn when Ashton discovers one of her besties has gotten himself into trouble with the not so human kind of blood-sucker.
Instead of letting the good times roll, she now finds herself heading into the heart of New Orleans.
And when preternatural havoc breaks loose, she will have to use her hidden powers and reveal her dark secret about what she truly is in order to survive.
Blood Meridian is an exciting, action packed, melanin-magic infused, urban fantasy, paranormal thriller.
COMING LATER IN 2021 TO RETAIL PLATFORMS WHERE BOOKS ARE SOLD
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October 13 2016Bob Dylan Wins Nobel Prize in Literature Bob Dylan, the songwriter known for a wide-ranging career and complex, symbolic, and often esoteric lyrics delivered in a raspy, mercurial lilt, has won the 109th Nobel Prize in Literature. The award was announced on October 13 by Sara Danius, permanent secretary of the Swedish Academy.
September 12 2016Pew Report: Americans Still Love Libraries, Especially the Books More than half of all Americans 16 and over used a public library in the past year, either in person or via the Web, according to a survey report on library use released this week by the Pew Research Center. The survey also found that Americans continue to view public libraries as vital to their communities: some 77% say that public libraries provide them with the resources they need, and 66% say the closing of their local public library would have a major impact on their community.
August 17 2016How Canada Publishes So Many Diverse Children’s BooksLibrarians wandering the publisher displays at the big U.S. conferences often seem surprised to discover that so many of their favorites are Canadian. But word has gotten out. Groundwood Books, which released Sidewalk Flowers, was named the best North American children’s publisher of 2016 at the Bologna Book Fair, the largest international trade show dedicated to children’s books. The award is for distinguished creative and publishing excellence. In fact, three of the five publishers nominated were Canadian. Quietly but surely, publishers in Canada are building a worldwide reputation for originality, quality, and diversity in children’s literature.
August 03 2016Oprah's Book Club Pick: The Underground RailroadOprah Winfrey has chosen Colson Whitehead's The Underground Railroad as the latest Oprah's Book Club Pick. She told the Associated Press "that she knew from the first sentence that she would want to share her passion with her audience, an impulse she has relied on with uncommon success for 20 years."
July 27 2016The Couple Next Door by Shari LapenaWatch for this suspense debut by Shari Lapena from Penguin Random House. Coming in August and already is getting great marketing buzz. Also a Dewey Diva pick.
June 13 2016Review: Annie Proulx's Barkskins“Annie Proulx’s Barkskins is remarkable not just for its length, but for its scope and ambition. It’s a monumental achievement, one that will perhaps be remembered as her finest work."
Create a new kind of library experience for your patrons, with tools from technology to decor.
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Encephalitis mosquito (Japanese, aestivo-autumnal) — the acute neuro and viral illness with a transmissible way of transfer proceeding as a pan-encephalomeningitis.
Etiology. The activator — the RNA-genomic virus relating to an arbovirus is a little steady in external environment, is inactivated by disinfecting solutions, termolabilen; it is well reproduced on the chicken embryos intertwined in cultures of cells of the person and animals. White mice, hamsters, monkeys and other animals are sensitive to infection.
Epidemiology. The illness belongs to natural and focal (endemic) infections. The reservoir — wildings, birds. A carrier — a mosquito, transovarial transfer is possible. Seasonality — aestivo-autumnal. The person and domestic animals can be an infection source. In Russia is registered in areas of the southern Primorye.
Pathogeny. Infection occurs at a sting of a mosquito. The virus extends on all organism gematogenno, possesses a neyrotropnost. The virus is reproduced in neurons with parenchymatous degeneration, is more often in the field of subcrustal nodes, a mesencephalon, a hypothalamus. Brain covers are involved in process.
Clinic. Incubation interval of 5 — 14 days. The beginning is acute, temperature is up to 40 °C within 7 — 10 days. The first days — a fever, a headache, an ache, nausea, vomiting, a muscular hypertension. The next 5 — 10 days — the expressed meningeal syndrome, motive concern, symptoms of oral automatism, orientation disturbance, aggression, nonsense. In cerebrospinal fluid lymphocytes 0,2 • 109/l. In hard cases develop heavy diffusion encephalitis (encephalomyelitis), up to a cerebral coma, trunk frustration, the central paresis and paralyzes in different combinations, alalias, defeats of kernels of cranial nerves. In blood — a leukocytosis with a neutrocytosis. The lethality is noted in 70% of cases.
Reconvalescence is long. From the residual phenomena in the forefront mental disorders, motive disturbances are expressed more weakly.
Differential diagnosis. Mosquito encephalitis should be differentiated with primary and secondary encephalitis, its initial stage (1 — 3 day) — with flu at which are observed pain in the field of superciliary arches, the catarral phenomena, a tracheobronchitis, in blood — a leukopenia.
Sluggish paralyzes of a shoulder girdle are characteristic of a tick-borne encephalitis at which mental disorders are much less often observed, seldom there are a hypertension of muscles, spastic paralyzes. Besides, various seasonality, carriers, endemicity is considered epidanamnez. Differentiation with other arbovirusny encephalitis (horse, San Louis, Scottish, etc.) is carried out on the basis of epidemiological Data (different areas of distribution), results of virologic and serological researches.
Hemorrhagic fevers sometimes proceed with the encephalitis phenomena. Damage of kidneys (nephrosonephritis), rash, a heavy hemorrhagic syndrome allow to exclude mosquito encephalitis.
Epidemic encephalitis (Ekonomo) differs from mosquito in the general constraint, drowsiness, existence of okulo-lethargic and vestibular syndromes. At it the loss of consciousness, a meningeal syndrome are seldom observed. The residual phenomena — parkinsonism, at mosquito encephalitis — psychoses, decrease in intelligence, the general adynamy.
Secondary encephalitis owing to adeno-and an enteroviral infection, flu, parotitis, chicken pox, measles and other viral infections differs in dominance of the all-brain phenomena (a gipertenzionny syndrome, wet brain), focal defeats are, as a rule, poorly expressed and are not followed by persistent widespread paresis. Besides, or objectively find the symptoms characteristic of the corresponding infection in the anamnesis of an illness. At secondary viral encephalitis the leukopenia with a relative lymphocytosis is observed, at mosquito — a leukocytosis with a neutrocytosis.
The two-wave current, the catarral phenomena or diarrhea during the doparalitichesky period, development of sluggish paralyzes are characteristic of poliomyelitis, is more often than the lower extremities. Children of younger age are ill mainly.
The diagnosis podtverzhatsya by allocation of a virus from cerebrospinal fluid, blood of patients, a brain of the dead, intratserebralny or intraperitoneal infection of white mice. The virus is identified in a neutralization test and RTGA. Serological diagnosis — 4-fold increase in pair serums of antibodies in RTGA, RSK or a neutralization test.

domain: health


And it's not just the Web. Graphical word processors and quick graphics programs have made it easy for everyone to create amazing visual effects in seconds.
- "Pretty presentation is more important than a good message."
This isn't really something people say out loud very often. But it dominates a lot of decision making in business. People re-design their web sites again and again. They choose "ooooo" and "aaaaaah" over content and usability.
A few posts back I talked about business cards. Flashy and useless wins the day over practical. But this issue goes way beyond business cards. Awhile back I was listening to a presentation by Michelle Ponto, a local TV news producer in Sacramento. She was talking about how to get the attention of the local media.
One of her key points is that press releases should be simple and well written. Avoid graphics. Avoid fancy fonts. Avoid anything that can't be skimmed FAST for the news team. In other words: If you make their job easier, your chances of getting coverage go up.
Content is king.
Another example is Google. The Google home page is approximately the most elegant and useful page on the Internet. And for all the billions of sites that compete for first place, the simple Google home page is number one.
There's always a fight of form versus function. But when you live in a technical world, remember that function needs a little extra attention. Web pages, press releases, resumes, and job proposals are all read by computers first and people second. In many cases, these communications are read by thousands of computers and very few people!
You don't have to eliminate the look and feel of your communications. But give the computers their due.
This blog has been criticized at times because it doesn't follow any of the norms for success. My posts are long, they're almost all text with few pretty pictures. Theoretically, no one should be reading this. But you are. Thank you.
There are no current plans to add a video a day, a bunch of moving graphics, slide show galleries, or a spoken advertisement when you hit the page.
Pretty is good. But a good message is better.
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Animal welfare charity Battersea has praised the Scottish Government and MSPs from across Parliament for delivering on their promise to increase maximum sentences for the worst animal cruelty offences.
The Animals and Wildlife (Penalties, Protections and Powers) (Scotland) Bill first introduced in October last year, passed its third and final reading on June 17.
For Battersea, this saw the culmination of many years of campaigning to raise Scotland’s maximum sentences to five years, in line with many other countries in Europe. The charity first launched its campaign to increase maximum sentencing in Scotland in 2017 and have been encouraged by the Scottish Government’s commitment to this change in law.
This Bill will raise the maximum sentence for animal cruelty in Scotland from 12 months to five years, and will become the law after it receives Royal Assent.
Research published by Battersea in 2017 showed that the maximum sentence of 12 months for animal cruelty was among the lowest across the whole of Europe – with only five other countries having lower sentences (including England and Wales). Both fly tipping and theft carried higher sentences.
Battersea has been calling on the Government in England and Wales to follow Scotland’s example. The Bill has been published twice before but fell during the prorogation of Parliament in October 2019, and a second time when a general election was called in December.
A Private Member’s Bill brought in by Chris Loder MP is currently seeking to introduce these measures, but has now been delayed until October 23, five months after its original date for discussion in Westminster.


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…we have been vindicated-Transparency International
By Ayo Fadimu
There is palpable panic across the country as the Federal government announced plans to investigate and prosecute the saboteurs of its Social Investment Programmes (SIPs).
One particular programme of SIPs, Tradermoni had generated several complains of fraud and underhand antics where beneficiaries were shortchanged to as much as 20 per cent of the loan given by the Federal Government
TraderMoni is a collateral-free loan targeted at petty traders and artisans, is part of the National Social Investment Programme, under the Government Enterprise and Empowerment Programme, through which the Federal Government says it intends to bridge the credit gap and empower Nigerians at the grassroots.
Beneficiaries of TraderMoni, which is coordinated by the Bank of Industry and the Government Enterprise and Empowerment Programme, a unit in the Office of the Vice President, have six months to repay the loan with flexible options of N85 daily or N430 weekly.
The beneficiaries, who receive an initial loan of N10,000, will qualify to obtain further loans of N15,000, N20,000 and N50,000, if they don’t default in repaying the facility.
However, in a statement by Communications Manager, National Social Investment Office (NSIO), Justice Bibiye,
on Sunday in Abuja, the Special Adviser to the President on Social Investments, Mrs. Maryam Uwais said the Federal Government has concluded plans to prosecute saboteurs of the program
She said the Federal Government would continue to mainstream the principles of transparency, accountability and probity in the implementation of the SIPs of the President Muhammadu Buhari administration. She said efforts were being intensified to track down those bent on sabotaging the SIPs by exploiting beneficiaries for personal gains, or sensationalise untruths to discredit the efforts at uplifting poorer citizens of the country from poverty.
Uwais said NSIO was working with the relevant security agencies to ensure that those found wanting were arrested and prosecuted. She responded against the backdrop of media reports which quoted her to have accused monarchs and other traditional rulers in the country of collecting bribes from beneficiaries of the SIPs.
In the meantime, Uwais has aide urged members of the public to dismiss such reports as sensationalism and a total misrepresentation of facts. Uwais delivered a goodwill message as Special Guest at the 13th Anti-corruption Agenda for the 9th National Assembly, held recently in Abuja. She said she never mentioned at that event that monarchs or traditional rulers as among those involved in demanding kickbacks from SIPs beneficiaries in rural areas.
According to Uwais, her comments at the function, organised by the Human and Environmental Development Agenda, a Non-Governmental Organisation, were based on the outcome of investigations triggered by an individual in the audience. She said that the concerns raised at the event related to complaints by some beneficiaries who alleged being short-changed by some community leaders in Osun State.
“At a similar event held sometimes ago, a young man had publicly reported that some beneficiaries were being shortchanged in his community, so I promised to have it investigated. “This investigation was carried out by ANEEJ, the principal monitor of disbursements of the Abacha restituted funds. “The report of the investigation revealed that the disclosures were essentially factual; some community leaders and LGA officials were in the habit of demanding levies after payment is concluded in the communities.’’ Uwais said that NSIO resolved to undertake measures to counter those adverse practices in the field. According her, the office is planning an event in the communities to engage the beneficiaries with a view to building their confidence, so they can refuse to accede to such demands. “Those who understand the leadership structure in our communities would know that monarchs are not the only set of persons classified as community leaders. “Our royal fathers are usually supported by others in the hierarchy to superintend over community affairs. “It then becomes very disturbing and somewhat mischievous for a reporter to single out monarchs from a comment that focused generally on community leaders.
‘We urge the media to avoid sensationalism and keep striving towards upholding the lofty principles of truth, fairness, objectivity and accuracy in their reportage of events of National concern. “In addition to field reports we get from our independent monitors and volunteers, we have continued to encourage and welcome feed-backs from members of the public, on acts of infraction and other challenges that can assist government to achieve better results in the implementation of the Social Safety Net Programmes,’’ She said
There have been reported cases of fraud in several states including Lagos, Osun, Kogi, Abia and Kaduna States among others. A claim supported by Transparency International
In an exclusive chat with Nigerian NewsDirect, the Head of Transparency International Nigeria, Auwal Musa Rafsanjani said the position of the government had vindicated the international body has it had raised alarm over the fraud being perpetrated in the scheme but the government was upset.
“Now that the government had found that there are fraudstersand saboteurs that are exploiting the beneficiaries, they should prosecute the offenders as announced” Rafsanjani told Nigerian NewsDirect.

domain: law


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You know what they say – everyone has a twin that looks just like them. While it’s rare for everyday folks like us to find our doubles, in Hollywood, celebrity lookalikes are not hard to come by. And it’s not because they have the same plastic surgeon in common.
They genuinely look alike. So much so, that you almost think these celebrities are related. But they’re totally not. And that’s what makes their striking resemblance so bizarre.
So, without further ado, we present to you 20 pairs of celebrities that look alike and could pass for twins. Side note: prepare to be mind-blown!
1. Emma Stone and Lindsay Lohan
Emma Stone and Lindsay Lohan truly are the mirror image of each other. It’s not just their ginger locks they have in common. Even as blondes, these actresses are absolute lookalikes.
From their identical eyes to their noses and lips, Emma and Lindsay are basically twins. Even Lindsay seems to agree! In 2017, the star tweeted Emma asking if she wanted to film Mean Girls 2 together (even though that terrible sequel already exists).
2. Tinashe and Jade Thirlwall
We’re convinced that Jade Thirlwall and Tinashe are the same person. The Little Mix singer and 2 On star look so much alike, it’s difficult distinguishing between the two.
They’re so similar, we can’t look at Jade without seeing Tinashe. Could it be that Jade has a secret solo career she’s told no one about? One time, she even sang along to Tinashe’s song and gave her a shout on Twitter by calling her “Sis”. What more proof do we need, ladies and gents?
3. Paul Wesley and Justin Bieber
Despite the 12-year age gap, it’s easy to believe that Paul Wesley and Justin Bieber must have been separated at birth. With similar eyebrows, eyes and noses, these two really are celebrity doppelgangers (Vampire Diaries fans – see what we did there?).
The Stefan Salvatore actor once joked that he was Bieber’s dad – and it’s totally believable. After E! posted a funny side-by-side photo of Paul and Bieber where they look alike, Paul wrote in the comments, “He’s my son.” #KnewIt
4. Jennifer Lawrence and Bella Hadid
We can’t help but see two lookalikes when Jennifer Lawrence and Bella Hadid are side by side. Whether it’s the shape of the eyes or cute lips, there’s just something about these celebs that is strikingly similar.
If they aren’t the same person, then Jennifer and Bella could totally pass for twins. Who knows? Maybe they are sisters who were separated at birth – and were both destined to hit Hollywood…
5. Kendall Jenner and Anya Taylor-Joy
We don’t know if you see it too, but Anya Taylor-Joy is literally Kendall Jenner‘s clone. Put dark hair on The Queen’s Gambit actress and we guarantee it will be a mirror image of the the Kardashian-Jenner model.
Whether it’s their rounded hazel eyes or their petite face shape, these two celebs must be sisters in an alternate universe!
6. Mark Ruffalo and Noah Centineo
These two actors may look like polar opposites considering their age difference, but look closely (especially at a younger Mark Ruffalo) and you’ll notice that they’re actually very similar.
Noah Centineo, the prince of modern romance movies, looks ever so similar to Hulk actor, Mark Ruffalo, from his face expressions to even his voice! In fact, Mark joked about the comparisons on The Tonight Show Starring Jimmy Fallon: “Wow. That’s amazing. He’s way better looking than I was…He might be able to play my dad one day.”
7. Margot Robbie and Emma Mackey
Emma Mackey’s mom clearly said “copy, paste” when she was born, because she is the doppelganger of actress Margot Robbie! These celebrity doppelgangers are so similar, it kinda freaks us out.
Unlike other celebs on this list, however, the Sex Education star doesn’t like being compared to Margot. “I genuinely just don’t see it at all,” she said. “Like, it’s lovely to be compared to Margot Robbie, but mostly I’d rather people focus on the jobs that we’re both doing rather than what we look like.”
8. Tobey Maguire and Elijah Wood
These blue-eyed actors are a reflection of each other in many ways from their wee lips to their pointy chins. Tobey Maguire and Elijah Wood are both renowned actors in their own right, but you have to admit it’s easy to confuse the two when you see them in their major blockbusters.
The Spider-Man actor and Lord of the Rings frontman are so alike, that Elijah admits to being confused for Tobey by a fan: “I’ve been mistaken for Tobey Maguire”, he revealed on Chatty Man. Funnily enough, it was such an awkward run-in that he never corrected the fan by telling her who he really was.
9. Katy Perry and Zooey Deschanel
This identical celebrity pair is pretty much common knowledge in Hollywood – everyone agrees that they’re practically one and the same. Katy Perry and Zooey Deschanel have such an eerie resemblance from their distinct blue eyes down to their gorgeous poses.
In fact, both these stars agree that they might be long-lost sisters, as Katy admitted to posing as the actress to get into clubs before she became a best-selling female artist. Zooey also starred in Katy’s Not the End of the World video, whereby she gets abducted by aliens who mistake her for the popstar. Even the aliens agree!
10. Robert Downey Jr. and Jeffrey Dean Morgan
Robert Downey Jr. and Jeffrey Dean Morgan are strikingly similar. Both of a similar age, these Hollywood hunks tend to confuse movie-goers around the world with their matching features.
Something else they have in common? They both have a thing for the fantasy genre – one’s a Marvel hero and the other is a Walking Dead character. Are we sure these lookalike stars aren’t secretly related?
11. Mila Kunis and Sarah Hyland
Mila Kunis and Sarah Hyland have such an uncanny resemblance, it’s difficult telling these actresses apart. Mila has the iconic big eyes – Sarah has the iconic big eyes. Even their lips and eyebrows are the same!
The TV stars themselves even admit their similarities. Sarah answered a tweet that pointed out their resemblance saying, “We know ;)”. And even Mila revealed that she often gets confused for the Modern Family actress. “You pretend to be me. Can I pretend to be you?’” Hyland replied to her.
12. Will Ferrell and Chad Smith
These celebrity lookalikes might just be the most shocking and accurate on our list. Comedy movie genius Will Ferrell and Red Hot Chili Peppers star Chad Smith are the absolute spitting image of each other.
These very obvious long-lost twin brothers often acknowledge that they are each other’s doppelgangers. They even appeared on The Tonight Show Starring Jimmy Fallon in matching outfits for an epic drum-off. How can two famous people look so much alike?!
13. Amy Adams and Isla Fisher
There’s no denying that Amy Adams and Isla Fisher are two of a kind. These Hollywood actresses look so alike, it’s easy to mix them up when they’re on screen.
Even Lady Gaga has awkwardly confused the two. Isla Fisher admitted to The Hollywood Reporter that the singer said: ‘Thank you. Your performance in American Hustle, Amy…” As a big fan of Gaga, she didn’t want to tell her that she wasn’t Amy Adams: “So I just gracefully thanked her and bowed my head.”
14. Harry Styles and Mick Jagger
From their similar faces to their unique rocker style, Harry Styles and Mick Jagger are clearly two peas in a pod. I mean, it’s not just the boyband status that they have in common, is it? These artists look incredibly alike!
Of course, Mick is a lot older now, and we’re comparing the young version of him to Harry. At least we know what the One Direction lad will look like when he hits his 70s though, right?
15. Zoe Saldana and Thandie Newton
Are we sure Thandie Newton isn’t Zoe Saldana’s older (and British) sister? I mean, she’s a dead ringer for the Guardians of the Galaxy star! Their signature cheekbones and petite frames certainly make it hard to tell them apart.
Fun fact? Zoe revealed that her mother regularly confuses her for Thandie. “My mom still thinks that I’m in Westworld,” she said on The Late Late Show.
16. Hilary Duff and Victoria Pedretti
When she starred in You, we always thought Victoria Pedretti reminded us of someone. Then she went blonde in The Haunting of Bly Manor and we realized who it was – none other than Hilary Duff.
While we all think these young actresses are the carbon copy of each other, Pedretti has admitted that she doesn’t agree: “I don’t really look like any of these women”, she said, when she was compared to Hilary Duff, Kat Dennings, Jennifer Garner, Hilary Swank, and others.
17. Brittany Murphy and Lili Reinhart
If you’re a Riverdale fan, there’s no doubt that you also done a double take when looking at Lili Reinhart. She’s the mini replica of the late actress Brittany Murphy!
“They think I’m her reincarnated”, Lili revealed. Well, we’ll be damned if you weren’t, because the similarities are insane!
18. Charlie Hunnam and Travis Fimmel
Both belong to two of the biggest TV series around and now we’re wondering if they belong to the same parents, too! Charlie Hunnam (Sons of Anarchy) and Travis Fimmel (Vikings) make us do a double take every time they’re on screen thanks to their twinning looks.
Let’s be honest. Has anyone actually seen Charlie and Travis in the same room at the same time? No? That settles the case then. They must be the same person.
22 Celebrities That Committed Crimes
19. Nicki Minaj and Cardi B
There will be a lot of fans who will fight us on this one. Nicki Minaj and Cardi B are often pitted against each other as female rap stars, but it’s clearly not just the music to blame – they also look so alike!
This isn’t something either rapstress would agree on though, as these celebrities have feuded in the past. That doesn’t take away the fact that they still look exactly the same. We said what we said.
20. Jessica Chastain and Bryce Dallas Howard
We won’t be convinced that Jessica and Bryce are two different people until we see them standing next to each other in front of our very own eyes. These actresses are such celebrity lookalikes to the point where it confuses us.
I mean, we’re not the only ones who think it. Bryce’s father, Ron Howard even mistook Jessica for his own daughter. Now, if that doesn’t convince you of their resemblance, we don’t know what will.
So, there you have it. 20 pairs of celebrities that look like each other. It’s safe to say the Hollywood world works in mysterious ways. It won’t be long before another celeb makes an appearance looking just like another famous face.
What did you think of these famous lookalikes? Let us know if you have any others in mind in the comments below…

domain: celebrity


This project is a collection of static corpora (plural of "corpus") that are potentially useful in the creation of weird internet stuff. I've found that, as a creator, sometimes I am making something that needs access to a lot of adjectives, but not necessarily every adjective in the English language. So for the last year I've been copy/pasting an adjs.json file from project to project. This is kind of awful, so I'm hoping that this project will at least help me keep everything in one place. I would like this to help with rapid prototyping of projects. For example: you might use nouns.json to start with, just to see if an idea you had was any good. Once you've built the project quickly around the nouns collection, you can then rip it out and replace it with a more complex or exhaustive data source.


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Aamir Liaquat accuses Imran Khan of trying to break his marriage
Stay tuned with 24 News HD Android App
Dr Aamir Liaquat Hussain—the controversial TV host —known for his double entendre statements, has been under the scrutiny of the netizens due to rumors of third marriage failure story.
Who would have thought that the newlywed couple would come up with one of the biggest conspiracies being told on internet that former prime minister Imran khan was allegedly behind the rumors of their divorce.
Aamir got divorced by his second wife Syeda Tuba Anwar and tied the knot with Dania Aamir.
Dania Amir posted video to shoot down the rumours and said herself and Amir Liqauat are together and there is no separation. She appealed people to stop such rumours.
پی تی آئی والوں تم میرے گھر میں گھسے ہو مجبور مت کرو کہ بنی گالہ میں تیسری بیوی کے قصے سناؤں۔۔ میں ایف آئی آے اورُمتعلقہ اداروں سے مطالبہُ کرتا ہوں ان نام نہاد صحافیوں اور سوشل میڈیا کے اکاؤنتس چلانے والے جھوٹوں کو گرفتار کریں جو ہماری خدانخواستہ علیحدگی کی خبریں پھیلا رہے ہیں pic.twitter.com/efShsLItXS— Aamir Liaquat Husain (@AamirLiaquat) April 12, 2022
Amir Liauqat warned people of PTI that they have allegedly tried to mud slugging news about his house.
He said that PTI workers should not force him to tell the story of third wife in the Bani gala.
Amir Liqauat said urged the FIA and its affiliates to arrest the so-called journalists and liars running social media accounts who are spreading the news of separation with his new wife.
The controversial king of the television said, “I vehemently deny all the rumors that PTI and its mercenaries are spreading that I have parted ways with Dania. We love each other immensely. I warned PTI and their rented goons., Don't enter in my home, otherwise nothing will be left.”
I vehemently deny all the rumors that PTI and its mercenaries are spreading that I have parted ways with Dania. We love each other immensely. I warned PTI and their rented goons., Don't enter in my home, otherwise nothing will be left@ImranKhanPTI pic.twitter.com/xXBmpVa4wb— Aamir Liaquat Husain (@AamirLiaquat) April 12, 2022
In another tweet, Amir Liaquat urged PTI chairman Imran Khan not to bring rumours about his separation otherwise he would make statements too that would become unbearable for him.
واللہ عمران خان میرے گھر تک مت آؤ میرا اللہ کے سواُکوئی مدد گار نہیں اگر تمہارے حقائق پر ٹوٹ پڑا تو بات بہت آگے نکل جائے گی !! @ImranKhanPTI pic.twitter.com/hYTZKDWFio— Aamir Liaquat Husain (@AamirLiaquat) April 12, 2022
You can read below the public comments on the Dania Amir clarification video.


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This morning I started working on the paintings I was supposed to be done last week, but i was scraping paint off, feeling like I was going backwards, worried I'm not going to have enough ready for The Artist Project, but it had to be done.
Here's one of them, Scraping done, adjustments made, now I'm back to where I wanted to be. Just the foreground trees to go:
- Posted using BlogPress from my iPhone
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domain: artistic


Unfortunately, due to circumstances beyond his control, Matt Cranitch has had to postpone his advertised talk in the Handed Down series in Scartaglin and it has been rescheduled into next season’s run.
The Handed Down event on Saturday, January 27th has been given the working title: Creating a Sliabh Luachra Archive – Preserving Our Identity and it will be presented by Maggie Prendiville-Keane.
Kerry County Council Grant
“We were recently awarded a grant from Kerry County Council to help set up an archive of Sliabh Luachra material,” said PJ Teahan by way of an explanation of the title.
“We have been collecting for years and at present have a large collection of rare audio, video and photographic material which we want to digitize and make available to the public.
History of the Music
“Through the lecture series, Handed Down we have examined many topics relating to our area and in doing so have introduced the history of the music to new generations.
“We’re aware of our responsibilities in using public funding. At Handed Down on Saturday, January 27th., local historian Maggie Prendiville-Keane will reveal some of these rare pieces to show the community what exists and what we envisage the Handed Down / Sliabh Luachra Archive will become,” said PJ.
Items already in the collection includes unreleased recordings from a variety of sources.
From 1973 comes a recording of Paddy Cronin playing with Paddy Glackin on the street in Listowel just after the latter had won the senior fiddle All Ireland title there.
An audio recording from Tom and Mary McCarthy’s Bar, Castleisland in 1962 recorded for Radio Éireann by Andreas Ó Gallachóir.
Dan O’Connell memorial Award
Also on Saurday, January 27th. the Handed Down team will be presented with the Dan O’Connell Memorial Award.
To mark the occasion Jackie Daly has been invited to attend and play for a few parts of a polka set.
Groups or individuals wishing to dance are welcome and among the former dancers invited are: Timmy the Brit, Ann Keane and Eily Buckley, Aileen Cronin and the O’Connell family from Dan O’Connell’s Bar.
Guests on the night will be: Jackie Daly, Dave Buckley, and musicians: Mike Rice; Gearoid Keating and Mairead Corridan, Margaret B. Looney, Billy Joy and friends.
Filling the young musicians slot on the night will be: Meabh Ní Chonaill, Lauren O’Connor and Caoimhe Murphy.
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