Balancing Aromatherapy Candle by Neal’s Yard
Neal’s Yard have fused their expertise of natural skincare with home fragrance, to create heavenly handmade candles. Organic wax is combined with a soothing blend of bergamot and patchouli to relieve anxiety and tension. Burn after a frantic day, safe in the knowledge that no harmful toxins are being released.
Plantscription Anti-Ageing Eye Treatment by Origins
Gentle, yet potent plant extracts, Anogeissus Tree Bark and Kombucha, help to reduce signs of aging, around the delicate eye area. The cream gel formula absorbs quickly, lifting and smoothing the skin, which is up to three times thinner than facial skin. Collagen levels are also boosted to improve elasticity dramatically.
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IL18RB is an alias for the gene, Interleukin 18 receptor accessory protein, in humans. Other names include: Interleukin-18 receptor accessory protein; IL-18 receptor accessory protein; IL-18RAcP; Accessory protein-like; AcPL; CD218 antigen-like family member B; CDw218b; IL-1R accessory protein-like; IL-1RAcPL; Interleukin-1 receptor 7; IL-1R-7; IL-1R7; Interleukin-18 receptor accessory protein-like; Interleukin-18 receptor beta; IL-18R-beta; IL-18Rbeta; CD antigen CD218b; CD218b. The encoded protein has an amino acid length of 599 and a mass of 68.3 kDa. Other isoforms may exist. IL18RB is a member of the Interleukin-1 receptor family. This gene shares homology with other species, including: Mouse.


With a perfected blend of nourishing ingredients and essential oils it melts into hands leaving them silk smooth, hydrated and delicately scented. The combination of the sweet and juicy aroma of pear with the delicate, floral fragrance of freesia. This creates a superbly uplifting scent to revive your senses. Essential oils and nourishing ingredients are combined to create a collection that softens the skin and leaves it delicately perfumed.
Once you’ve tried Naturally European Hand Cream we’re sure you’ll agree it’s the best hand cream around for soothing dry hands.
Fortified with 20% shea butter, it’s thick texture melts into hands leaving them smooth and softly scented.
See our complete range of The Somerset Toiletry Company products.


EY-B002203Regular price $5.99 Sale price $1.00 Save 83%
Made of food-grade silicone, BPA-free materials ensure food safety for you.
The applicable ambient temperature for the silicone mold is 60 DEG C ~: 230 DEG C, Can be utilized in the microwave oven, grill, heating plate, Can be frozen in the refrigerator and freezer as well, With the good thermal conductivity to ensure the best effect of cooking and baking, This non-stick silicone mold features particularly easy mold unloading, which is smooth and almost perfect.
Silicone Chocolate Molds is soft and non-stick, silicone candy molds let you take out them easily and without damaging delicate food. Easy to fill and clean, dishwasher safe.
This cake mold can not only make round cakes, mousse cakes, tiramisu, but also bread, pudding, jelly, chocolate, etc. More features are waiting for you to discover.
Color: As shown in the figure
Temperature: -40F to 446F (-40°+230 )
Commercial level quality, durable, reusable, non-stick
Microwave and freezer safe
Easy to use and clean
Suitable for cake, muffin, chocolate, jelly, ice cream, soap, etc
1 * Cake Silicone mold


A high intake of milk won’t increase cholesterol levels, according to a large study of 2 million people.
In fact, those who drank milk every day had lower levels of bad (LDL) cholesterol than non-milk drinkers.
Moreover, higher intakes of milk were associated with a 14 percent reduced risk of coronary heart disease and an 11 percent reduced risk of type 2 diabetes.
Increased consumption of high-fat dairy, due to their saturated fat content, may have negative effects on cardiovascular health.
Saturated fats have been associated with high blood cholesterol and cardiovascular disease (CVDs) risk.
However, several studies have found that consumption of milk and low-fat dairy products lead to lower cholesterol levels and lower risk of CVDs.
This study examined the effect of milk intake on cardio-metabolic risk (risk factors for developing cardiovascular diseases and diabetes) using a genetic approach by looking at the lactase gene variant.
Lactase is a digestive enzyme which breaks down lactose, a sugar found in milk and milk products.
The enzyme is encoded by the lactase (LCT) gene.
The team found that people who drink higher amounts of milk have the genetic variation that digests lactose.
Professor Vimal Karani, the study’s lead author, said:
“We found that among participants with a genetic variation that we associated with higher milk intake, they had higher BMI, body fat, but importantly had lower levels of good and bad cholesterol.
We also found that those with the genetic variation had a significantly lower risk of coronary heart disease.
All of this suggests that reducing the intake of milk might not be necessary for preventing cardiovascular diseases.”
The reasons that high milk intake can lower cholesterol levels might be related to:
- Calcium and lactose in milk influencing fat metabolism.
- Compared to other dairy foods, milk is low in fat but people with lactose intolerance, to get their calcium intakes, tend to eat high fat dairy products which contain higher amounts of cholesterol.
- Calcium intake from foods such as milk enhances bile acid removal and so causes hepatic cholesterol to generate bile acids, consequently this process leads to a reduction of cholesterol levels.
- Gut microbial fermentation of indigestible carbohydrates such as lactose can affect cholesterol production causing cholesterol levels to drop.
Professor Vimal Karanis aid:
“The study certainly shows that milk consumption is not a significant issue for cardiovascular disease risk even though there was a small rise in BMI and body fat among milk drinkers.
What we do note in the study is that it remains unclear whether it is the fat content in dairy products that is contributing to the lower cholesterol levels or it is due to an unknown ‘milk factor’.”
The study was published in the International Journal of Obesity (Vimaleswaran et al., 2021).


Thanks so much for submitting a report. It has been emailed to the Rum Captain and will be actioned shortly.
Tasty, but not quite great
Smooth, sweet and soothing a bit classier than kraken, much cheaper which is a bonus. Love the cola finish and sweetness is just about right especially on a long warm summer evening.
Very sweet and heavy on the vanilla. Drinking neat has a bit of alcohol bite.
Similar to The Kraken but for me this is a bit better than that.
Creamy. Heavy on vanilla. A nice allspice burn. Finishes out with a little kola nut. Surprised me.
I actually prefer Black Magic to The Kraken--way cheaper, and doesn't feel like drinking Dr Pepper mixed with Everclear. This rum is very sweet, with lots of caramel and vanilla. The finish is also much smoother than Kraken. For two-thirds the price, it's a better choice.
I open the bottle and the nose was very over powering of sweetness.. similar to butterscotch. The taste was ok, it was smooth in mix, and did its job.. I'm particular to kraken and the taste, but this would do on a airplane ride in crunch time.
Similiar label and advertising, similiar product. Syrupy, very sweet, flavors of vanilla, root beer and chocolate. Aroma is swiss miss hot chocolate. Not great, not awful. Makes a really sweet rum and coke. Not sure what I'm going to do with this bottle.
"Packed with sweet flavours. It also has a very healthy base of sugar clearly added to it. Which is smoothing out the flavour of this rum."
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Chipotles are actually smoked dried jalapeños. We used traditional Mexican recipes to guide us in making this sauce as authentic as possible with our local ingredients.
This sauce is packed with a natural smokiness which adds to the flavor of this sauce.
No liquid smoke used, hours was spent smoking the Jalapeños on a closed braai with wine barrel oak shavings.
1 Year shelf life


The rise in antibiotic-resistant bacteria is a major concern, in particular because it includes many different species of pathogenic microbes. These “superbugs” are further characterized by high levels of virulence and disease-associated mortality. There seems to be few new antibiotics in the drug discovery pipeline; recent work has sought to define and validate new drug targets. The assembly of surface proteins and pili in the cell wall envelope of Gram-positive bacteria is catalyzed by sortase. Sortase cleaves a conserved C-terminal sequence of these polypeptides to generate an acyl-enzyme intermediate. The acyl-enzyme is next resolved by nucleophilic attack by the amino groups within cell wall cross-bridges or pilin proteins, thereby covalently attaching the polypeptides to the cell wall or the next pilin subunit. Sortase substrates function as adhesins, internalins, blood clotting and immune evasion factors, and transporters for nutrients across the microbial cell wall envelope; without them, most pathogens cannot sustain an infection. Here we review what is known about sortase catalysis and surface protein function, how surface protein anchoring can be inhibited, and what prospects such inhibition may have for anti-infective therapy.
I. Targeting Virulence Factors as a Therapy for Bacterial Infections
Infectious microbes can be viewed as an endless tide, challenging individual human lives as well as entire populations and providing strong selective forces for evolution (Krause, 1981). There are many different outcomes to the endless number of encounters that occur between an individual and its pathogens, most of which shape innate and acquired immune responses and affect future encounters with the same pathogen. Until the first half of the twentieth century, infectious diseases were a major factor in determining the average life span in human populations (Krause, 1981). Therefore, it is not surprising that even the earliest experimental work on microbial infections was concerned with prevention or therapy of these diseases (Ehrlich, 1911; Koch, 1882). Several landmark discoveries over the course of the last century identified natural compounds derived from microbes (e.g., sulfonamides, penicillin, and streptomycin) that block replication of sensitive bacteria (Amyes, 2001). Such antibiotics not only transformed the diagnostic and therapeutic practices of the medical profession, they also expanded human life expectancy. Alongside a flurry of discoveries in the field of microbial antibiotics, the pharmacological industry achieved development of synthetic compounds that selectively block bacterial replication, thereby expanding the register of chemicals that can function as magic bullets. These widely celebrated research accomplishments instilled broad confidence into regulatory agencies as well as practicing physicians that all bacterial infections can be controlled by antibiotic treatment (Stewart, 1967). Such ambition is now broadly challenged by the emergence of microbial strains that are resistant to many, if not all, antibiotics currently in clinical use. Thus, the recent pressing challenge of microbial research is the identification of novel drug targets and the development of inhibitors that can be used for therapy of human infections.
The emergence of microbial strains resistant to antibiotics is a phenomenon of selection. Massive production and use of these compounds in industry and the clinic are not regulated and represent the basis for emergence of antibiotic-resistant strains. Considering the constellation of drug discovery, production, distribution, and use, the medical profession is engaged in an arms race because new antibiotics must be discovered to avoid therapeutic failure due to selection pressure of compounds currently in use (Bonomo, 2007). However, the generation of new antibiotics must be accompanied by an identification of novel bacterial targets whose inhibition would be of therapeutic value. The corollary to this is to develop anti-infectives that inhibit bacterial virulence strategies, such as attachment to host tissues, for example. Assuming that anti-infective therapy of this nature can be implemented for bacterial infections, such compounds may not be associated with the same resistance phenomena as antibiotics with bactericidal activity, a conjecture that remains speculative because such therapy has not been developed or approved for human use. At the outset, there are limitations to targeting virulence factors as possible therapies. Even if inhibited in their virulence strategies, pathogenic microbes require opsono-phagocytic killing by the immune system for host clearance, which would limit the use of such therapies to immune-competent individuals. Furthermore, this strategy requires knowledge of the invading pathogen, and any therapy would have to be delayed until a definitive identification of the pathogen has been made. In summary, the inhibition of virulence factors as a therapy for bacterial infections still represents a theory that requires experimental testing and proof-of-principle. Here, we discuss sortase, a prominent virulence factor in Gram-positive pathogens, as a target for the development of anti-infectives and examine recent research progress toward this goal.
II. Staphylococcus aureus
A. Clinical Disease and Epidemiology
Staphylococcus aureus is a Gram-positive, catalase-positive bacterium that colonizes human skin and mucosal surfaces. The microbe is isolated from at least 40% of the human population (Lowy, 1998, 2003; Peacock et al., 2001). Skin or soft tissue infections typically result from local breaches in epithelial barrier functions and may be resolved locally or lead to rapidly spreading infections with involvement of any internal organ system (Lowy, 1998). If sensitive to antibiotics, S. aureus infections can be effectively treated with β-lactam antibiotics, macrolides, and fluoroquinolones (Lowy, 2003). Staphylococcal resistance to β-lactam antibiotics is now widespread. Within one year of the introduction of methicillin, a nonhydrolyzable synthetic β-lactam, methicillin-resistant S. aureus (MRSA1) strain carrying the mecA gene and catalyzing peptidoglycan transpeptidation, even in the presence of antibiotic, was isolated from clinical infections (Jevons et al., 1963). The incidence of MRSA infection is increasing throughout the world, accounting for significant mortality and associated health care costs (Klevens et al., 2006, 2007; Noskin et al., 2007). Vancomycin and other glycopeptide inhibitors were once considered last resort therapies for MRSA; however, strains with intermediate or full resistance to vancomycin have been described previously (Chang et al., 2003; Weigel et al., 2003). Perhaps even more alarming is the rise of community-acquired MRSA strains (Klevens et al., 2007). Community-acquired -MRSA infections occur in children with no predisposing risk factors, young adults (including athletes, homosexual men, prisoners, and military recruits), and healthy newborns (Fridkin et al., 2005). Collectively, S. aureus precipitates up to 3 million clinical infections each year in the United States (Fridkin et al., 2005). For both community- and hospital-acquired S. aureus infections, antibiotic-resistant strains represent a formidable and increasing therapeutic challenge (King et al., 2006). It is estimated that up to 100,000 Americans will succumb to MRSA infections this year (Fridkin et al., 2005; King et al., 2006). Even with expert medical care, these highly destructive and debilitating infections can be fatal unless diagnosed early and treated aggressively (Gillet et al., 2002).
B. Staphylococcal Virulence Genes
The development of animal models for infection together with molecular biology techniques permitted studies on the genetic basis for staphylococcal diseases. We now know that S. aureus requires a plethora of virulence factors to establish an infection. S. aureus strains secrete more than fifty polypeptides that enable bacterial escape from innate immune responses, promote tissue and cellular damage, or degrade connective tissues (Sibbald et al., 2006). The aggregate of all exoprotein secretion events provides for staphylococcal spread in infected tissues (Novick, 2003b). Expression of many exoprotein genes (and therefore the secretion of encoded polypeptides) is controlled by agr, the staphylococcal quorum sensor and regulator of virulence genes (Recsei et al., 1986; Ji et al., 1995). Other important regulatory and sensory molecules include two-component systems (SaeRS) and transcription factors (Sar) (Cheung and Zhang, 2002; Novick, 2003a; Steinhuber et al., 2003). It is noteworthy that deletion of a single exoprotein gene typically does not cripple the resulting mutant strain or diminish its virulence properties. An important exception to this notion is the secretion of α-hemolysin, a pore-forming toxin that is essential for the pathogenesis of experimental lung infections in mice (Wardenburg et al., 2007). However, mutations that abrogate the function of regulatory factors involved in the coordinated expression of exoproteins (agr, saeRS mgrA, or sar) cause virulence defects in almost every animal model system examined, indicating that many exoproteins fulfill redundant yet essential functions (Cheung et al., 1994). In agreement with this hypothesis, pioneering studies by Richard Novick, Tom Muir, and colleagues developed inhibitors of Agr function that display therapeutic effects when added to skin infections (Lyon et al., 2000). These compounds do not affect staphylococcal growth in laboratory media but demonstrate therapeutic efficacy and can therefore be considered as examples of compounds that target virulence strategies.
The most extensive knowledge for animal models that develop invasive staphylococcal diseases has been accumulated in mice. Intraperitoneal or blood stream infection of mice with many different human clinical S. aureus isolates, typically at a dose of 1 to 10 × 106 colony-forming units, is followed by opsono-phagocytic killing of up to 99% of invading staphylococci within 1 to 2 h (Albus et al., 1991; Mazmanian et al., 2000). Approximately 103 to 104 staphylococci escape phagocytic killing, exit the bloodstream, and adhere eventually to various organ tissues. Within 3 to 4 days, staphylococci replicate to large numbers and are sequestered by immune cell-mediated liquefaction necrosis (Bubeck Wardenburg et al., 2006). Staphylococcal abscesses harbor central lesions filled with staphylococci and cellular debris that are surrounded by large numbers of polymorphonuclear cells and macrophages (Bubeck Wardenburg et al., 2006). Peripheral fibrin deposits confine abscesses from other tissues and restrict fluid circulation into the lesion (Bubeck Wardenburg et al., 2006). The elucidation of abscesses in infected animals is often used as a marker for the severity and progression of staphylococcal disease (Albus et al., 1991; McKenney et al., 1999; Mazmanian et al., 2000).
C. The Staphylococcal Cell Wall Envelope
The cell wall of staphylococci and other Gram-positive bacteria represents a surface organelle composed of a primary polymer, peptidoglycan, and secondary polymers (protein, carbohydrates, and teichoic acids) that are immobilized in the peptidoglycan scaffold. In S. aureus, the 30 to 100-nm thick cell wall is composed of the repeating disaccharide N-acetylmuramic acid-(β1–4)-N-acetylglucosamine (MurNAc-GlcNAc) (Ghuysen and Strominger, 1963; Ghuysen et al., 1965; Dmitriev et al., 2004). MurNAc is amide-linked to alanine of the cell wall tetrapeptide [l-Ala-d-isoGln-l-Lys(NH2-Gly5)-d-Ala], which is linked to adjacent strands of tetrapeptide through a pentaglycine cross-bridge (Fig. 1) (Ghuysen et al., 1965; Tipper and Strominger, 1965; Tipper et al., 1965; Tipper, 1968; Archibald, 1972). This cross-linking gives rise to the murein sacculus, a single large macro-molecule that completely envelopes the cell (Beveridge, 1981). In addition to imparting shape and architecture, the cell wall contains covalently and noncovalently associated proteins involved in adhesion, nutrient sensing/uptake, immunomodulation, and the transfer of DNA (Moks et al., 1986; Flock et al., 1987; Chen and Cleary, 1990; Galli et al., 1990, 1992; Gaillard et al., 1991; Clewell, 1993; Burne and Penders, 1994; Patti et al., 1994; Dramsi et al., 1995; Herwald et al., 2004). Furthermore, it can be decorated with (lipo)teichoic acids, teichuronic acids, and polysaccharide (Robbins and Schneerson, 1990; Fischer, 1994; Neuhaus and Baddiley, 2003). During infection, the sum of all cell wall polymers enables many of the pathogenic strategies of S. aureus.
D. Surface Protein
Sequence analysis of some known surface proteins in Gram-positive bacteria identified a conserved LPXTG motif (Fischetti et al., 1990). The LPXTG motif is one element of cell wall-sorting signals positioned at the C-terminal end of precursor surface proteins, which contain also a hydrophobic domain and positively charged tail (Schneewind et al., 1993). Disruption of two or more of the positively charged residues leads to secretion into the media, suggesting that the polybasic region of surface proteins acts as a retention signal for cell wall anchoring (Schneewind et al., 1993). Moreover, fusion of the sorting signal to the C-terminal end of precursor proteins with N-terminal signal peptides directs their hybrid product to the cell wall envelope (Schneewind et al., 1992; Navarre and Schneewind, 1994, 1996). Chemical analysis of anchored polypeptides liberated from the cell wall envelope by enzymatic cleavage of peptidoglycan fragments revealed that the C-terminal end of surface protein, the threonine of the LPXTG motif, is amide-linked via its carboxyl group to the amino group of the pentaglycine cross-bridge in the cell wall (Schneewind et al., 1995; Ton-That et al., 1997; Navarre et al., 1998, 1999). A screen of mutant populations of S. aureus led to the identification of the sortase gene, which is responsible for the anchoring of surface proteins to the cell wall envelope (Fig. 2). Since this discovery, hundreds of homologs of the sortase gene have been identified, encompassing ∼900 potential substrates in over 50 different species of Gram-positive bacteria. Sortase-like genes have even been identified in Gram-negative organisms (Pallen et al., 2001, 2003; Comfort and Clubb, 2004). In most cases, multiple sortases are found in the same genome and can be grouped based on their homology into four or five classes (Comfort and Clubb, 2004; Dramsi et al., 2005). Sortase A is the prototypical enzyme for members of the class A family, the so-called “house-keeping” sortase. One of the shared features of sortase A members is that they are responsible for the anchoring of proteins that mediate bacterial adhesion. Members of this class of sortase seem to be ubiquitously expressed. Class B sortases are regulated by the availability of iron and anchor proteins that are involved in heme-iron acquisition. Class C sortases are responsible for the elaboration of pili on the bacterial surface. Finally, class D sortases are expressed during bacterial sporulation, and their anchored products contribute to the developmental programs that define the life cycle of bacilli or streptomyces (Fig. 3). Throughout this review, unless a specific sortase (i.e., sortases A–D) is mentioned, we use the term sortase to refer to the general properties of this class of transpeptidases using sortase A as a prototype.
A. Sortase-Catalyzed Transpeptidation
S. aureus sortase A is a 206 amino acid protein with an N-terminal signal peptide/membrane anchor. Replacement of its N-terminal 25 residues with an affinity tag provides for a single step purification strategy of a soluble hexahistidyl-tagged enzyme on nickel-nitrilotriacetic acid-Sepharose from lysates of recombinant Escherichia coli that express srtA under control of the T7 promoter. The substrate abz-LPETG-dnp harbors an N-terminal fluorophore (2-amino-benzyl) and a C-terminal quencher (2,4-dinitrophenyl); fluorescence resonance energy transfer (FRET) occurs in the substrate due to the close proximity of fluorophore and quencher. Sortase A-mediated cleavage precipitates separation of reaction products, abz-LPET and G-dnp, and fluorescence increase serves as a measure for sortase cleavage of surface protein substrates (Ton-That et al., 1999, 2000). Addition of hydroxylamine (NH2OH), a strong nucleophile, to this reaction increases the velocity of sortase cleavage both in vitro and in vivo. Sortase forms surface protein LPET-hydroxamate in vivo, indicating that NH2OH attacks the acyl-enzyme intermediate (Mazmanian et al., 1999; Ton-That et al., 2000). Similar to hydroxylamine, peptidoglycan substrates and their surrogates, for example, Gly, Gly2, Gly3, Gly4, or Gly5 (i.e., the pentaglycine cross-bridge), also stimulate sortase cleavage of abz-LPETG-dnp. The products of this reaction, abz-LPET-Gly, abz-LPET-Gly2, and so forth, document that the amino group of the cell wall cross-bridge performs the nucleophilic attack at the acyl-enzyme intermediate of sortase A both in vivo and in vitro (Mazmanian et al., 1999; Ton-That et al., 1999, 2000). When offered a peptidoglycan substrate with amino group nucleophile, sortase A invariably functions as a transpeptidase but not as a peptide hydrolase. The physiological nucleophile for the sortase A-catalyzed anchoring reaction is lipid II, the peptidoglycan biosynthesis precursor [C55-PP-MurNAc(-l-Ala-d-isoGln-l-Lys(NH2-Gly5)-d-Ala-d-Ala)-GlcNAc], because this compound can be linked to surface proteins both in vivo and in vitro (Perry et al., 2002; Ruzin et al., 2002). Vancomycin, which binds to lipid II and prevents its incorporation into the cell wall, and moenomycin, a lipid II analog, block cell wall synthesis and the sorting pathway of surface proteins (Ton-That and Schneewind, 1999).
Separation of sortase reaction products by reversed phase-high-performance liquid chromatography and mass spectrometry of eluate fractions demonstrated that LPETG is cleaved between the threonine and the glycine. These experiments also revealed the formation of the acyl-enzyme intermediate, i.e., the thioester linkage between LPET and the active site cysteine thiol of sortase A in vitro (Huang et al., 2003). The addition of [2-(trimethylammonium)ethyl]methanethiosulfonate and p-hydroxymercuribenzoic acid, two sulfhydryl-modifying agents, to staphylococcal cultures or to the in vitro sortase reaction each inhibits substrate cleavage by modifying the active site cysteine (Ton-That and Schneewind, 1999; Ton-That et al., 1999). Iodoacetamide, a compound that reacts rather slowly with thiol, cannot inactivate sortase. One explanation for this observation may be that the cysteine of sortase is ionized (Ton-That and Schneewind, 1999). In agreement with the proposed role of the sole cysteine residue in catalysis, substitution of Cys184 with alanine abolishes sortase A activity both in vivo and in vitro (Ton-That et al., 1999, 2000). Bioinformatic analysis revealed that the active site cysteine is a conserved feature of sortases, in agreement with the general hypothesis that mechanisms of surface protein anchoring to the cell wall envelope are conserved in Gram-positive bacteria (Ton-That et al., 1999).
B. Sortase Structure and Catalysis
NMR and X-ray crystallography of S. aureus sortase A revealed that the enzyme adopts a unique eight-stranded β-barrel, which contains several short helices and loops (Ilangovan et al., 2001a,b; Zong et al., 2004a). The active site is located in a hydrophobic depression formed by two β-stands, features that are preserved in sortase B from S. aureus and Bacillus anthracis. Two conserved residues, His120 and Arg197, are positioned in close proximity to the active site sulfhydryl of Cys184 (Ilangovan et al., 2001b; Zong et al., 2004a). Cocrystals of sortase and LPETG revealed that Cys184 and Arg197 reside between the side chains of the scissile peptide bond, suitably positioned to act as a catalytic cysteine-arginine dyad, where Arg197 may function as a base for thiol ionization (Zong et al., 2004a). Both His120 and Arg197 are involved in catalysis; however, their mechanistic contributions are still under investigation (Ton-That et al., 2002; Conolly et al., 2003; Dessen, 2004; Marraffini et al., 2004; Zhang et al., 2004; Zong et al., 2004b; Frankel et al., 2005). It seems unlikely that the catalytic contributions of His120 and Arg197 can be resolved until detailed structural appreciation for binding of peptidoglycan and polypeptide substrates to sortase is achieved. Using a cyanoalkene analog of LPETG and NMR analysis, three residues within the hydrophobic compression adjacent to the active site (Thr180, Ile182, and Ala118) were demonstrated to undergo rearrangement as the enzyme engages its substrate. In agreement with the hypothesis that Thr180, Ile182, and Ala118 are involved in recognition and stabilization of bound substrate, amino acid substitution of each of these residues can have profound effects on sortase activity (Liew et al., 2004). Calcium ions stimulate S. aureus sortase A activity 8-fold but have little or no impact on the activity of S. aureus sortase B or B. anthracis sortase A, sortase B, or sortase C (see below). Calcium concentrations required for stimulation of sortase A activity are found under physiological conditions in host tissues, and these ions are involved in structural rearrangements of a disordered loop covering the active site that enable substrate binding (Ilangovan et al., 2001b; Zong et al., 2004a; Naik et al., 2006). Interestingly, sortase also forms dimers in vitro, a function that may stimulate catalysis (Lu et al., 2007).
C. The Pathway of Surface Protein Anchoring
A model illustrated in Fig. 2 assimilates many of the observations summarized here for S. aureus sortase A. Surface proteins are synthesized in the bacterial cytoplasm as precursors harboring an N-terminal signal peptide and a C-terminal sorting signal (P1 precursor). After initiation of surface proteins into the Sec secretion pathway, signal peptidase cleaves the N-terminal signal peptide to generate the P2 precursor. The hydrophobic domain of the sorting signal is thought to retain the polypeptide in the membrane, thereby allowing the LPXTG motif to be recognized by sortase A. The sulfhydryl of Cys184, presumably activated for nucleophilic attack via deprotonation by Arg197, forms a covalent bond with the carbonyl carbon of threonine at the C-terminal end of the cleaved polypeptide, releasing the amino group of the cleaved C-terminal fragment. The product of the first part of the sortase-catalyzed transpeptidation reaction, the thioester linked acyl-enzyme intermediate, is subject to nucleophilic attack by the amino group of the pentaglycine cross-bridge within the peptidoglycan precursor lipid II and thereby forms the P3 intermediate [C55-PP-MurNAc(-l-Ala-d-isoGln-l-Lys(surface protein-Gly5)-d-Ala-d-Ala)-GlcNAc]. P3 intermediates are subsequently incorporated into the envelope via the transpeptidation and transglycosylation reactions of cell wall biosynthesis, ultimately generating mature product (M), i.e., surface proteins that are covalently linked at the C-terminal end to the cell wall envelope.
D. Sortases, Surface Proteins, and the Pathogenesis of Microbial Infections
Interest in sortase as a target for the development of anti-infectives primarily stems from findings that loss of the sortase gene leads to defects in the pathogenesis of many different infections caused by Gram-positive bacteria. For example, S. aureus virulence can be assessed in a mouse model of infection (Lee et al., 1987; Albus et al., 1991; McKenney et al., 1999). When injected at a sublethal dose into the blood stream of mice, most staphylococci survive innate immune response, disseminate to peripheral tissues and establish abscesses in multiple organ systems over 72 to 96 h. The products of the infection can be quantified as bacterial load in homogenized tissue (colony-forming units) or via histopathology for their anatomical substrate. In these models, srtA mutants display a 2-log reduction in bacterial growth in multiple organ systems as well as a 1.5-log increase in the lethal dose (LD50) compared to wild-type strains (Mazmanian et al., 2000). These findings prompted further investigations into the contribution of S. aureus sortase A to other types of infection, including septic arthritis and endocarditis (Bremell et al., 1991, 1992). In the septic arthritis model, staphylococcal blood stream inoculations lead to joint infections with cartilage and bone destruction, which can be assessed by histopathology. In experimental endocarditis, lesions on the heart valves serve as a site of deposition of staphylococci, causing destructive damage of endocardial tissue. In both model systems, srtA mutants displayed large reductions in pathogenesis (Jonsson et al., 2002, 2003; Weiss et al., 2004). Recently, an experimental model of S. aureus lung infection was developed, which monitors the development of acute pneumonia and disease-associated mortality following intranasal inoculation of mice with staphylococci. Animals infected with srtA mutants show less disease than their wild-type counterparts (Wardenburg et al., 2007). Finally, the ability of surface proteins to illicit protective immunity following vaccination with purified components was assessed for Group B streptococcal, pneumococcal, and staphylococcal infections. By combining sortase substrates as vaccine antigens that generated the highest protective immunity, animals could be protected against lethal challenge with the pathogen. For example, the combination of IsdA, IsdB, SdrD, and SdrE as vaccine antigens protected mice against lethal challenge with S. aureus strains that had been isolated from human clinical infections, including MRSA strains USA100, USA300, and USA400, the most common cause of infections in the United States (Stranger-Jones et al., 2006). Virulence attributes of sortase and surface protein extend well beyond staphylococci, as defects in the pathogenesis of sortase mutants have been reported for animal infections with many different species, including actinomyces, enterococci, streptococci, bacilli, or listeria (Bierne et al., 2002; Garandeau et al., 2002; Kharat and Tomasz, 2003; Lee and Boran, 2003; Chen et al., 2005; Lalioui et al., 2005; Levesque et al., 2005; Paterson and Mitchell, 2005; Sabet et al., 2005; Yamaguchi et al., 2006). Collectively, these studies illustrate the many important and diverse roles that surface proteins play during the pathogenesis of infections caused by Gram-positive pathogens.
E. Iron Acquisition
Pathogenic bacteria have an absolute requirement for iron as a nutrient during the infectious process (Crosa et al., 2004). S. aureus sortase B (SrtB) recognizes a surface protein with an NPQTN sorting signal, which plays a role in iron acquisition (Mazmanian et al., 2002). In S. aureus, srtB is located in the Isd locus; name was given because it contains genes whose products are all involved in heme-iron scavenging and transport. Specifically, the locus encodes cell wall-anchored heme-binding proteins (IsdA, IsdB, and IsdC), a membrane-based heme transport system (IsdD, IsdE, and IsdF), a heme degrading monooxygenase (IsdG), and SrtB. The locus is regulated by the ferric uptake regulator protein, a repressor of DNA transcription at the promoter of target genes during iron-replete conditions (Mazmanian et al., 2003; Skaar et al., 2004). IsdC appears to be the only surface protein substrate in S. aureus that is anchored by SrtB, which cleaves the substrate at its NPQTN motif-sorting signal and immobilizes it at cell wall cross-bridges. IsdA and IsdB, however, are anchored to the cell wall by sortase A (Mazmanian et al., 2002). In contrast to IsdA and IsdB, IsdC remains buried within the staphylococcal cell wall envelope, presumably because it is linked to mature assembled peptidoglycan but not to lipid II. As a consequence, IsdC is not incorporated into linear peptidoglycan strands that eventually reach the envelope surface, unlike all sortase A-anchored products (Mazmanian et al., 2002; Marraffini and Schneewind, 2005). The aforementioned results were also the first indication that sortase genes, which are positioned in the same transcriptional unit as putative surface proteins, generally anchor these substrates to the envelope. Loss of srtB reduces both staphylococcal growth on heme as the sole source of iron and transport of heme across the cell membrane, implicating sortase B function in the pathway whereby staphylococci acquire iron during infection. In the renal abscess model, srtB mutant staphylococci show defects in the persistence of bacterial infections and an overall decrease in virulence in a murine infectious arthritis model, corroborating the hypothesis that heme uptake is important during the infectious process (Mazmanian et al., 2002; Jonsson et al., 2003; Weiss et al., 2004). SrtB homologs are also found in other Gram-positive pathogens, including Listeria monocytogenes and B. anthracis. In listeria, SrtB surface anchors two proteins, SpvA and an IsdC homolog. B. anthracis IsdC, which is also a substrate of SrtB, binds heme and is required for growth on heme as the sole source of iron (Bierne et al., 2004; Pucciarelli et al., 2005; Maresso and Schneewind, 2006; Maresso et al., 2006; Skaar et al., 2006). As iron acquisition is essential for bacterial replication during infection, inhibition of sortase B offers an opportunity to disrupt infectious processes.
F. Sortase and Pili
Pili represent some of the most important virulence determinants for bacterial infection of a mammalian host. These macro-molecular structures often consist of repeating protein subunits, which extend from the bacterial surface into the surrounding medium. The tip or cap of these fimbriae typically display adhesive properties that promote bacterial binding to extracellular matrices or host cell receptors in the context of an infection. The first indication that sortase may anchor components of this structure was revealed in actinomyces, pathogens responsible for tooth decay (Yeung and Ragsdale, 1997; Yeung et al., 1998). Reconstitution of immunoreactivity in a heterologous host (E. coli) using anti-pilin antisera allowed for the identification of an operon, which contained a pilin-like gene and a gene later characterized as sortase. Mutagenesis of the putative sortase abolished actinomyces adherence and led to the loss of pili from the bacterial surface (Yeung and Ragsdale, 1997; Yeung et al., 1998).
Definitive evidence that some sortases polymerize pili in the cell wall envelope occurred with the discovery of six sortase-like genes in Corynebacterium diphtheriae,a human respiratory pathogen that causes diphtheria (Hadfield et al., 2000). Specific surface proteins or pilin subunits encoded by the same operon as sortases identify specific pilus types, of which C. diphtheriae expresses at least three. For example, antibodies against the SpaA protein (sortase-mediated pilin assembly A), a component of one of these loci, stained fimbrae-like structures in a uniform fashion, whereas those against SpaC, a second pilin component of the locus, stained the tip of pilus fibers. A third component, SpaB, is also distributed along the fiber shaft. Deletion of both the spaA and srtA genes abolished formation of SpaA pili, whereas deletion of spaB and spaC abrogated only binding of the corresponding pilin-specific antibodies without affecting assembly of SpaA pilin into pili by their cognate sortase. Furthermore, as assessed by immunoblotting, only srtA and spaA mutants failed to form high molecular weight species consistent with pilus assembly (Ton-That and Schneewind, 2003). Assembly was also dependent on a conserved lysine residue of SpaA, the critical residue in the “pilin motif”, which in conjunction with the sorting signal is sufficient for the formation of pili in a SrtA-dependent manner. The requirement of lysine is explained by the presumed role of its ϵ-amino group in relieving pilin acyl-enzyme intermediates with sortase. If this is the case, class C sortases may polymerize pili by forming covalent bonds between individual pilin subunits, and the bonds involve the cleavage site of pilin precursors as well as the pilin motif lysine residue. Class C sortases and their corresponding pili have been discovered in Actinomyces viscosis, Actinomyces naeslundii, Bacillus cereus, Clostridium perfringens, Streptococcus agalactiae, Enterococcus faecalis, Streptococcus pneumoniae, and Streptococcus pyogenes (Dramsi et al., 2005). Following the assembly of the pilus by SrtC, it is hypothesized that SrtA catalyzes the covalent attachment of the full-length structure onto the peptidoglycan (Swaminathan et al., 2007)
G. Sortase and Sporulation
B. anthracis harbors three sortase genes, srtA and srtB, with properties similar to those described for S. aureus, and a third gene, srtC (originally called srtC but is actually a member of the srtD class), in a locus that also encodes a two-component regulatory system (sctR and sctS) as well as basI (an LPNTA motif-containing protein) (Fig. 3) (Dramsi et al., 2005; Marraffini and Schneewind, 2006). Another LPNTA-motif containing protein, basH, is located elsewhere on the B. anthracis genome. Purified SrtC cleaves LPNTA between its threonine and alanine residue but does not cut the recognition motifs of sortase A (LPXTG) and sortase B (NPKTG), suggesting that both BasI and BasH may be in vivo substrates for sortase C. Using transcriptional fusions of srtC to gfp, srtC expression was observed only during sporulation and was dependent on the response regulator sctR. BasH-GFP hybrid expression was observed only in the developing forespore and was dependent on the presence of the transcription factor σF RNA polymerase. Analysis of cell wall fragments demonstrated that BasH and BasI are anchored to the forespore and mother cell peptidoglycan, respectively, and that both are dependent on a functional srtC gene, whose product is localized to the mother cell and the forespore envelopes. Deletion of the srtC gene caused no defect in virulence in B. anthracis strain Ames and carcasses of animals infected with the mutant teamed with vegetative forms, similar to carcasses derived from infection with the wild-type parent strain. However, vegetative forms of the srtC mutant were unable to form spores in carcass tissue, even 3 weeks after animal death, whereas wild-type bacilli formed spores within 24 h. A similar phenotype was observed in sheep's blood, as srtC mutants were unable to sporulate unless blood was rotated and exposed to oxygen. A general model was derived from these observations, whereby the sortase C-mediated anchoring of BasI and BasH to the cell wall envelope was critical for the generation of spores in host tissues (Marraffini and Schneewind, 2006). Although it is unlikely that this class of sortase would be specifically targeted for anti-infective development, given that spore formation is not usually observed during human infection, we should not discount the possibility for infection control after addition of sortase inhibitors to foodstuff contaminated with spores.
IV. The Inhibition of Sortase
A. Early Observations
The first inhibitors of the cell wall sorting reaction were discovered even before sortase was identified (Ton-That and Schneewind, 1999). At that time, it was known for S. aureus that surface proteins are linked through an amide bond to the amino group of pentaglycine cross-bridges (Ton-That et al., 1997, 1998; Navarre et al., 1998). The similarity of this mechanism to the transpeptidation reaction of cell wall biosynthesis led to the investigation of antibiotics as inhibitors of the sorting pathway. Penicillin, an inhibitor of the transpeptidation reaction of cell wall synthesis, had no effect on the generation of mature anchored surface proteins; however, vancomycin and moenomycin reduced surface protein anchoring. As vancomycin and moenomycin both target steps in the lipid II biosynthesis cycle, it was presumed that lipid II may be a substrate for the sorting reaction. Interestingly, compounds that are reactive with sulfhydryl groups, such as methanethiosulfonates, i.e., [2-(trimethylammonium)ethyl]methanethiosulfonate, were also inhibitors (>2 log units) of surface protein anchoring. p-Hydroxymercuribenzoic acid, which inhibits cysteine proteases, also inhibited anchoring, whereas iodoacetamide and DTT did not (Ton-That and Schneewind, 1999). These findings not only implicated sulfhydryl (cysteine) as a catalyst for surface protein anchoring but also demonstrated that inhibition of sortase in vivo could be measured.
B. Natural Products
After this discovery and the cloning of sortase, there have been several investigations for sortase inhibitors in compound libraries of natural and synthetic products (Table 1). Taking the approach that plants may harbor antimicrobial compounds, extracts of 80 medicinal plants from Korea were examined for inhibition of recombinant sortase (Kim et al., 2002). Cocculus trilobus, Fritillaria verticillata, Liriope platyphylla, and Rhus verniciflua extracts had the best inhibitory activity with IC50 values ranging from 1.5 to 8.4 μg/ml. Further purification of F. verticillata extracts led to the isolation of β-sitosterol-3-O-glucopyranoside as the active sortase inhibitor with an IC50 of 18.3 μg/ml, two times lower then p-hydroxymercuribenzoic acid (Kim et al., 2003). It is not known why crude extracts display a lower IC50 than the purified inhibitor. Elimination of the glucopyranoside side chain moiety led to a ∼1.5 log reduction in inhibitory activity, indicating that this moiety may be the active component of the compound. This compound also inhibited Bacillus subtilis and S. aureus growth with minimal inhibitor concentrations at 50 and 200 μg/ml, respectively. Bactericidal activity in a sortase inhibitor would indeed be a surprising feature, and it seems more likely that these compounds block other reactions that are essential for bacterial survival and growth on laboratory media. A simple test for the possibility that sortase inhibitors display antibiotic properties may be to determine the plating efficiency of wild-type and sortase mutants in the presence of compound. Assuming that sortase inhibitors perturb envelope functions in a manner that affects bacterial growth, such phenomena could not occur in strains lacking sortases. Until proven experimentally, it seems more likely that β-sitosterol-3-O-glucopyranoside exhibits cytotoxic effects on bacterial cells that are distinct from the inhibition of sortase.
Isoquinoline alkaloids (berberine chloride) from rhizomes of Coptis chinensis were also found to be inhibitors of sortase (IC50 of 8.7 μg/ml) and exert antibiotic activity (Kim et al., 2004). Further examination of marine invertebrate extracts (Topsentia genitrix), which are known to contain antimicrobial activity, lead to the identification of bis(indole) alkaloids of the topsentin class as sortase inhibitors with an IC50 of ∼15 μg/ml. One of these, bromodeoxytopsentin, was evaluated for its ability to inhibit S. aureus adherence to fibronectin, a proposed method for evaluating sortase inhibition in vivo. At approximately the IC50 in vitro, near complete inhibition of adherence to fibronectin was achieved, with 50% inhibition of adherence occurring at 2.5 μg/ml (Oh et al., 2005). Considering the IC50 in vivo is approximately six times better than the IC50 in vitro, it seems likely that there are nonspecific effects on adherence that are not related to the inhibition of sortase. Measurements of bacterial adherence to fibronectin are not a direct measurement of sortase activity, and these assays suffer from pitfalls associated with inhibition of other pathways that also effect adherence (e.g., inhibition of protein synthesis or secretion of polypeptide through the Sec secretion pathway). Curcumin and derivatives from Curcuma longa also inhibited sortase with appreciable activity (IC50 13–32 μg/ml) and potentially prevented adherence to fibronectin (Park et al., 2005). In similar fashion, inhibitors of SrtB from S. aureus were isolated from the bark of the plant R. verniciflua. These inhibitors, identified as flavonoid phenols, showed appreciable in vitro inhibition of sortase in the low micromolar range (Kang et al., 2006). It is noteworthy that three flavonoids, morin, kaempferol, and galangin, along with the already identified β-sitosterol-3-O-glucopyranoside, displayed at least a 3-fold difference in inhibition between SrtA and SrtB. These results suggest that it may be possible to identify inhibitors that discriminate between specific members of the sortase family, highlighting discrete differences in active site architecture between these enzymes. Preliminary structure-activity relationships suggest that the co-occurrence of a hydroxyl group at C-3 of ring C and meta-hydroxy groups at C-2′ and C-4′ of the B ring is required for appreciable sortase inhibition. Furthermore, many of these compounds seemed to effect the clumping of S. aureus cells to one another. Collectively, these studies represent the first steps toward the identification of inhibitors of sortase from natural sources and have generated compounds that warrant further development.
However, care must be taken in considering in vitro inhibition solely on the basis of IC50 values (Copeland, 2005). A determination of the mechanism of inhibition of sortase by these classes is required to completely understand their relative effectiveness and specificity; i.e., is inhibition reversible/irreversible or mechanism-based? What are the affinities and kinetics of inhibition? Does inhibition occur through interaction with active site residues, or is this elsewhere in the polypeptide? Is the inhibition competitive in the presence of substrate? In light of the fact that many compounds can inhibit enzyme activity based on redox action, promiscuous aggregation, denaturization, pH changes, and so forth, not to mention possible quenching of fluorescence or spurious reaction with peptide substrate when considering the sortase activity assay, it seems prudent to carefully investigate the nature of the inhibition (Seidler et al., 2003; Kruger et al., 2004b).
C. Rational Design
Several investigators have taken advantage of the active site thiol features of sortase or the conserved structure of its peptide substrate and designed inhibitors in a rational manner. The first such report investigated the use of synthetic substrate-derived inhibitor sequences with reactive electrophiles aimed at irreversibly modifying the active site thiol of sortase (Scott et al., 2002). Using the substrate recognition motif of staphylococcal SrtA (LPXTG), the authors replaced the scissile peptide bond between the Thr and Gly with diazoketone (-COCHN2) or chloromethyl ketone (-COCH2Cl). Both compounds displayed micromolar inhibitor constants (Ki ∼2.0 × 10-7), with the chloromethyl ketone demonstrating faster kinetics of inactivation. Along these lines, vinyl sulfones, electrophilic inhibitors of cysteine proteases, were examined for inhibition of sortase (Hanzlik and Thompson, 1984; Frankel et al., 2004). Collectively, these compounds displayed time-dependent irreversible inhibition of sortase. One compound, 3,3,3-trifluoro-1-(phenylsulfonyl)-1-propene, was most effective with an IC50 of 190 μM and a single-step inhibition mechanism, most likely due to its highly electrophilic character relative to the phenyl vinyl sulfone-related compound. Mass spectrometry confirmed covalent bond formation with the active site (Cys184) of SrtA for one member of this class of inhibitors, thereby confirming their reactivity to nucleophilic cysteines. In addition to substrate-derived inactivators, inhibitors aimed at imitating the transition state of the sorting reaction were evaluated for activity (Kruger et al., 2004a). A nonhydrolyzable phosphinic peptidomimetic (NH2-YALPE-Ala[PO2H-CH2]Gly-EE-NH2) showed competitive inhibition with an IC50 of 10 mM. Although poor in inhibition, the usefulness of this reagent was realized when a kinetic analysis of the inhibitory pattern suggested Ping-Pong Bi Bi hydrolytic shunt kinetic model for sortase catalysis (Kruger et al., 2004a). For a detailed description of the proposed kinetic mechanisms of sortase catalysis, the reader is referred to several thorough studies on this topic (Kruger et al., 2004b; Frankel et al., 2005, 2007). Substrate analogs can also be used to map peptide-binding regions on sortase. The partial amino acid sequence of the SrtA substrate LPAT fused to cyanoalkene and sulfhydryl moieties allowed for the identification of the hydrophobic surface between the β4 and β7 strands as substrate-anchoring regions (Liew et al., 2004). Others have used threonine analogs to covalently modify the active site of sortase (Jung et al., 2005).
D. High-Throughput Screen
With the increasing availability of small molecules, drug-like libraries, and robotic automation, the search for sortase inhibitors has now entered the era of high-throughput screening. A screen of 1000 diverse compounds for inhibition of sortase yielded a diarylacrylnitrile with an IC50 of 231 μM (Oh et al., 2004). Examination of the structure-activity relationships of this compound indicated placing the two benzene rings in the trans-orientation as a (Z)-diarylacrylonitrile lowered the IC50 to 28 μM. Further structure-activity relationship indicated that a 2,5-dimethoxy configuration was the most potent with a competitive inhibition profile. Dialysis and activity recovery experiments suggested that inhibition was reversible. Modeling studies suggested further that the phenyl rings of the inhibitor may interact with lipophilic residues of the sortase substrate binding pocket. Future work on this class of inhibitors will be needed to achieve a structural appreciation of sortase inhibition.
Another HTS for sortase inhibition used 135,000 compounds from a diverse small molecule library. Upon monitoring cleavage of the FRET substrate LPETG via an increase in fluorescence, approximately 6000 compounds displayed inhibitory activity compared to positive controls (no compounds added) (Maresso et al., 2007). Due to the high hit rate (∼4%) in this screen, a “drug-like” filter was applied to eliminate all hits that encompasses reactive, genotoxic, promiscuous (frequent hits in unrelated assays) compounds, or molecules that lacked drug-like properties. The remaining molecules were clustered via structural similarities and sampled on the basis of activity, properties, and potential to provide SAR. A final set of 407 compounds were subjected to secondary screens aimed at i) determining the inhibitory activity toward a structurally related thiol protease (papain) and ii) determining the broad-based inhibitory activity toward different sortase family members (B and C) and species (SrtA from B. anthracis). All compounds that showed appreciable inhibition of different sortases but not papain were considered to demonstrate specificity and were examined further. This selection process, i.e., imposing cheminformatic filters on HTS data sets, followed by confirmation assays and specificity constraints, allowed for the identification of some promising lead compounds (Fig. 4a).
Using this methodology, several unique classes of reversible and nonreversible sortase inhibitors were uncovered. One class, termed the aryl β-amino(ethyl) ketone (AAEK), was selected for further study due to its simple drug-like structure, marked selectivity, and inhibitor activity toward sortase (Fig. 4b). Inhibition with this class of compounds was time/dose-dependent, irreversible, and showed low micromolar IC50 and Ki values. Mass spectrometry confirmed that the inhibition was covalent; the inhibitor adduct differed from the parent compound in that the amino moiety had been lost during the reaction with sortase (Fig. 4b). This result suggested the AAEK class of inhibitors were mechanism-based inactivators, a type of covalent modifier that has clear precedence in on-market drugs (Copeland, 2005). A model was proposed and tested whereby an electrophilic olefin intermediate was generated following interaction with sortase, which reacted with the active site thiol. Cocrystallization of several AAEK family members with B. anthracis sortase B suggested that binding of inhibitors to the active site of sortase may first be mediated by an interaction of a charged ammonium species with an anionic pocket followed later by stabilization of the inhibitor in the pocket via stacking interactions between the aryl ring and an active site tyrosine (Fig. 4, c and d) (Maresso et al., 2007). Preliminary SAR-hinted anionic substituents in the para-position on the aryl ring are required for inhibition and lie in a cationic pocket above the site of reaction with the active site cysteine (unpublished observations).
V. On the Development of Sortase Inhibitors
To date, the search for inhibitors of sortases has involved natural, synthetic, and high-throughput screening methodologies. As the field moves toward the development of compounds that can be interrogated for therapeutic efficacy, several issues outlined below must be considered. Although still preliminary in nature, the chemical diversity of the already uncovered inhibitors of sortases provides important assets for future improvements of specificity and potency.
A. Assay Design
When offered a choice, drug developers prefer noncovalent inhibitors over those that modify enzymes covalently, with mechanism-based inactivation perhaps representing the exception to this rule (Copeland, 2005). Most of the inhibition studies described thus far utilize the FRET-based cleavage of LPETG as a measure of sortase activity. Although this assay is well suited for inhibitor identification via HTS (Z′ of 0.94), there are several drawbacks. For instance, the concentration of enzyme and substrate required to attain appropriate Z′ are high. Low catalytic turnover, combined with long incubation times, probably introduces bias toward selection of covalent inhibitors. This phenomenon has indeed been observed in our own studies (unpublished data) and may be circumvented by the addition of physiologic nucleophiles (i.e., pentaglycine of lipid II) to the reaction for greater activity at lower enzyme concentration. Of concern is also the bias toward thiol reactive compounds, because sortase utilizes a catalytic cysteine residue. A possible solution to such systematic error would be to perform sortase assays in the presence of reducing agent (e.g., dithiothreitol). Finally, the potential for fluorescence quenching or spurious reactions of compounds with peptide substrate and/or fluorophore must be examined with secondary assays that use different outputs for sortase activity (Kruger et al., 2004b).
B. In Vivo Evaluation of Sortase Inhibitors
It is clear that lead compounds identified by in vitro screens will have to be evaluated for in vivo inhibition of surface protein anchoring to the cell wall. One should be cautious in the evaluation of readouts that are not direct measurements of this process, such as adhesion of S. aureus to extracellular matrix proteins, for example. A plethora of mechanisms in protein secretion, folding, and anchoring to the cell wall envelope (including synthesis of peptidoglycan and other envelope components) affect the abundance of functional surface proteins in the staphylococcal envelope. A direct assessment of sortase activity is achieved by pulse-labeling polypeptides with [35S]methionine. Immunoprecipitation and separation of surface proteins on SDS-polyacrylamide gel electrophoresis followed by autoradiography can monitor the processing of pulse-labeled P1 precursor to P2 and mature species, thereby providing an in vivo measure for the sortase-catalyzed anchoring reaction. This time-consuming protocol is not useful for HTS; however, it can be used to characterize lead compounds. For HTS, we advocate the use of fluorescence-activated cell sorting (FACS) to monitor the abundance of anchored sortase substrates in the cell wall envelope. Finally, validation should include efficacy testing in animal models of infection that involve Gram-positive pathogens with sortases and surface proteins. In this respect, the commonly used renal abscess model appears suitable due to its large dynamic range (>3 log units) of virulence assessment, albeit that many other models may also be useful. However, it should not be forgotten that effective in vivo sortase inhibitors must also display favorable pharmacologic properties, including low toxicity and reactivity with mammalian proteases.
In an era of global spread of antibiotic resistance in many human pathogens, concerted efforts must be applied toward identification and validation of new targets for drug development. These include essential factors necessary for bacterial survival, even under nondisease conditions as well as virulence determinants whose actions are mostly manifested during the infectious process. Whereas antibiotics have a long-standing history of success in the treatment of bacterial infections, drugs that target virulence factors still suffer from a lack of experimental proof-of-principle. However, unless agents with novel inhibition mechanisms against essential factors are uncovered, the problem of resistance will continue. Sortase, a major virulence factor responsible for the covalent attachment of surface proteins to the cell wall, is a target candidate for the treatment of Gram-positive infections. Progress toward this goal has been made and several distinct sortase inhibitor classes have been identified. Compounds with reasonable inhibition, specificity, and mechanisms of inactivation have been uncovered. For these inhibitors to be realized as effective drugs, direct assessment of the inactivation of sortase and attenuation of virulence in animal models must be demonstrated. In a continuous struggle to stay ahead of the ever-changing drug resistance traits of pathogenic bacteria, the selective inhibition of virulence factors may prove to be an effective therapeutic strategy that may either augment or perhaps even supplant the more traditional approaches.
We apologize for the omission of any work that was not mentioned in this review due to space constraints. We acknowledge membership within and support from the Region V “Great Lakes” Regional Center of Excellence in Biodefense and Emerging Infectious Diseases Consortium (1-U54-AI-057153).
↵1 Abbreviations: MRSA, methicillin-resistant S. aureus; SrtA, B, and C, sortases A, B, and C; SpaA, sortase-mediated pilin assembly A; AAEK, aryl β-amino(ethyl) ketone; HTS, high-throughput screen; MurNAc-GlcNAc, N-acetylmuramic acid-(β1–4)-N-acetylglucosamine; FRET, fluorescence resonance energy transfer; Isd, iron-regulated surface determinant.
This work was supported by National Institutes of Health Grants AI38897, AI057153, and U54-AI-057153 (to O.S.) and National Research Service Award Grant AI069697 (to A.M.).
This article is available online at http://pharmrev.aspetjournals.org.
- The American Society for Pharmacology and Experimental Therapeutics


Types of medicinal formulations
In the pharmacy network, rhodiola rosea preparations are sold in the form tinctures, tablets or in the form of raw materials for the preparation of infusions, decoctions and teas at home.
- Infusion. This is a dosage form in liquid form, which is prepared according to a certain technology. The raw materials are poured with boiling water and allow time for the healing substances to pass into the liquid phase.
- Tincture. This is a liquid preparation of Rhodiola rosea in which alcohol is present. This form is stored for a long time without changing the quality.
- Decoction .. Golden root dosage form prepared by boiling dry roots in boiling water.
- Tea. The traditional method of preparing an aromatic invigorating drink.
The cure for hypotension - vodka tincture of Rhodiola rosea root
50 grams of the medicinal raw material in the form of well-dried roots is ground in a coffee grinder to a powder state. The resulting pollen is poured with vodka in the amount of two glasses. This mixture should be placed in a dark place for two weeks. While the tincture is preparing, it must be periodically brought into the light and gently shaken.
After 14 days, the finished remedy for hypotension is filtered and poured into a glass bottle of dark glass. To normalize the pressure at low tonometer, take the medicine two teaspoons before meals. The frequency of admission is three times a day. It is recommended to start taking the tincture of the golden root with a few drops and gradually bring it to the required volume.
The recipe for tincture of the root of Radiola pink for the treatment of nervous disorders, digestive disorders, colds
Two whetstones of dried roots are ground with a knife and brewed with a glass of boiling water. A mug with the future medicine is insisted for about four hours, then filtered through a gauze napkin. This is an effective rinse, a tool for washing wound surfaces. For the treatment of the above diseases, the infusion is taken one hundred grams three times a day.
The recipe for the golden root decoction for the treatment of diseases of the female and male genital organs
The decoction is used both externally for rubbing and lotions, and inward with exacerbation of pain, increased uterine bleeding, with impotence in men. When applied orally as an additional action, a golden root decoction reduces high blood sugar values, soothes a headache and pain in the lower abdomen. It can be used to rinse with diseases of the oral cavity - gingivitis, stomatitis and periodontosis.
To prepare the broth, boil water. It will take half a liter of boiling water. Grind four small sticks of dry roots and pour boiling water. The resulting mixture should be held in a water bath or low heat. The drug is insisted for a couple of hours. When the broth cools down, it is filtered and stored in the refrigerator. The dosage regimen is simple - 50 milliliters three times a day.
A decoction for treating sexual weakness, asthenia, neurosis and for recovering from a serious illness
This is a drug that strongly tones the central nervous system. Coffee or strong black tea does not give such a surge of vigor and strength. Such a potent drug is taken only if absolutely necessary. For cooking, take a spoonful of crushed raw material, which is poured with a liter of cool water. The mixture is brought to a boil and boiled for 10 minutes. The resulting broth is insisted for an hour, after having wrapped the dishes with the broth with a warm scarf. A tonic drink should be consumed throughout the day. To enhance the adaptogenic and stimulating effect, add a spoonful of honey or cane sugar to the broth.
The recipe for maintaining excellent immunity
This is a universal remedy for maintaining protective functions of the immune system. The mixture includes the fruits of hawthorn, rose hips, herbs that can be harvested in the summer or bought in a pharmacy. To prepare the mixture you will need such raw materials: Equal parts of 20 grams of dry rosehips, a golden root and a root of a zamani. To the collection obtained, add St. John's wort grass in the amount of 10 grams, nettle grass - 10 grams and 15 grams of red berries of hawthorn. A handful of the collection must be poured with boiling water and simmered for ten minutes over low heat. The resulting broth is insisted for several hours, then filtered. The resulting saturated liquid should be diluted with a glass of boiling water. The medicine is taken three times a day for 50-70 milliliters.


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The Secret Formula of Male enhancement pill® that Other Pills Don’t Have by ADMIN on JULY 9, 2010 Have you wondered why other male enhancement supplements fail to provide their users the expected results? Do you totally believe that overall virility can be achieved by taking pills with only a few ingredients in them? Men who want to gain the male sexual prowess the holistic way should consider being nourished by a supplement that gives all the benefits and effects naturally with combined powers of essential components. This is the main reason why other male enhancement pills fall short to their promise of providing the desired results to their users. However, with ExtenZe® Male Enhancement Supplement, a complete set of beneficial effects can be gained because of the secret formula that comes with every tablet. ExtenZe® is fortified with folic acid and zinc. How important these minerals are to the male enhancement process? Folic acid provides formation of healthy red blood cells and zinc integrates protection to their structure; thus, allowing increased resistance. Optimal blood flow is needed for penile tissue growth and development as well as to correct any vascular problem that may cause erectile dysfunction.


Having a cafe at a gas station or a c-store is vital for modern gas stations. While the competition grows tougher, fuel sales are decreasing. For this reason, gas station owners are looking for alternative sources of revenues.
This article was written specially for the leading industry publishing”Toplivnye Rynki”. Imagine a gas station of the future. How would it look like? What services should it provide? Whatever you say, the gas station of the future is impossible without retail operations and convenience services.


Bibliorganics Spicy Coconut Cider Vinegar (750ml)
- Unlike other synthetic vinegars in the market, Bibliorganics Spicy Coconut Vinegar is a rich source of vitamins, minerals and amino acids, making it the perfect culinary condiment and health tonic.
- This natural vinegar is culled from the sap of the coconut tree whose roots extend deep into the earth, reaching seawater tables, absorbing more than 65 essential minerals from seawater.
- These nutrients are carried to all parts of the coconut through the sap, which is naturally aged and fermented to reach the desired acidity.
- The special blend of chili, garlic and ginger makes it even healthier and more delectable.
- Bibliorganics Spicy Coconut Vinegar also contains FrutooligoSaccharides (FOS) which stimulate the production of beneficial bacteria that aids in digeston, nutrient absorption & prevention of major diseases It is also very low on the glycemic index, making it a suitable condiment for diabetics.
- Organic and all-natural ingredients.
- Naturally aged and fermented.
- This product is best used for cooking as opposed to Cocowonder Coconut Cider Vinegar which is recommended as a health tonic.
How do we use this product?
A great pantry staple that can be used for cooking or as a dipping sauce.
It may also be diluted in water or mixed with other ingredients, such as oil in a salad dressing or marinade.
Naturally Fermented Coconut Sap, Chili, Garlic, Ginger.
Size - 750ml
Other Search Terms - coco cider, braggs, suka, sukang puti, spicy suka, sukang paumbong.
This is our favorite vinegar at home! Goes perfect with our fried dishes! It's sour with that extra spicy kick! 🌶
Thank you very much!
Thank you very much


leather and fabric sneakers color: blue light gray leather inserts rubber striped sole composition: leather and textile fibers exterior leather and textile fiber interior other materials sole. made in romania.
gray yellow and blue suede sneakers white sole. composition: leather textile fibers and other materials interior textiles rubber sole. made in romania


Recent Examples of chamomile from the Web
Makes 1 cocktail 6 ounces hot water 1 bag chamomile tea 1-3/4 ounces honeysuckle vodka 1 rock sugar swizzle stick Steep tea bag in hot water in a rocks glass or coffee mug.
Jack Weiss, President/Owner Coco Pazzo Restaurants in Chicago Best meal: Whole chicken and chamomileRestaurant: Roister, 951 W. Fulton Market, www.
Because Wylie Dufresne opened Du’s Donuts, with flavors like pomegranate sesame and grapefruit chamomile, in W’burg.
Creamy sprouted almonds made their debut on the Mission Chinese menu as part of a dish of napa cabbage steamed in chamomile and masala milk.
If your skin is irritated, douse it in a lotion or oil that has aloe in it to soothe and moisturize the bumps, like Johnson's Baby Oil Gel With Aloe & Vitamin E or Parissa Azulene Oil, which has a calming chamomile derivative.
Intended to be rolled directly onto achy areas, the formula combines 17 essential oils, including geranium and chamomile, which are thought to stimulate circulation, and capsaicin (the active compound in the chili pepper) for an analgesic effect.
From Ever Since Contour Serum, $95 This silky anti-aging serum is infused with green tea extract and chamomile, plus a dose of cannabinoid oil, formulated to smooth itty bitty fine lines and tighten saggy, puffy skin.
Put a few drops of essential oil on your pillow or in a diffuser, or drink some herbal tea, like peppermint or chamomile, which is naturally caffeine-free.
These example sentences are selected automatically from various online news sources to reflect current usage of the word 'chamomile.' Views expressed in the examples do not represent the opinion of Merriam-Webster or its editors. Send us feedback.
Origin and Etymology of chamomile
First Known Use: 13th centurySee Words from the same year
CHAMOMILE Defined for English Language Learners
medical Definition of chamomile
Seen and Heard
What made you want to look up chamomile? Please tell us where you read or heard it (including the quote, if possible).


Fish cutlets pike recipe fishcake, burgers
How to cook schuchi (fish) cutlets, burgers. Take the fresh fish (pike), it must be clean, wash and separate the fillets. Fish fillet in minced meat to grind. Soak white bread (a loaf is better) in the milk and add the minced fish meat, grind onion, garlic can taste and add it to the stuffing. Move to the same egg, pepper, salt, and mix the stuffing. Fry the grated carrot and chopped onion in vegetable oil and also add to the stuffing. All this mix. Heat the pan with vegetable oil and fry patties until rosy state from both sides. Tip: To make it easier to form the patties in the hands, arms periodically lubricated with vegetable oil. After roasting, meat balls are added to the pot, which adds a small amount of water (steam), and there is paryatsya meatballs over low heat until cooked. At the end of cooking in a pan add black pepper peas and bay leaf. You can also cook the tomato gravy and pour it into a kettle of steaming meatballs in tomato sauce for.


11 Best Hemp Paper Benefits and Uses
When you think of all the daily uses of paper, it’s hard to imagine a world where it doesn’t exist. But with the majority of the world’s paper currently being made from trees with unsustainable practices and chemically intensive processes, shifting to tree-free paper might be just what the world needs. The verdicts already out on how hemp is one of the most sustainable fabrics, but how can hemp paper pave a path for a more sustainable future?
WHAT IS HEMP PAPER?
Most conventional paper is made out of trees, but hemp paper is made from the long bast fibers or short bast fibers (hurd) of the hemp plant.
Are you wondering how to make paper from plants? Well, for hemp paper, the process is pretty simple. It involves breaking down hemp fibers into a pulp that can then be laid out and molded into a screen paper-making frame and dried.
Other than a couple of pretty major exceptions, the basics of how to make paper from plants aren’t that different for hemp and trees. These major exceptions are that hemp paper needs far less chemical processing than tree paper products, and tree paper has a huge negative environmental impact, while hemp paper is eco-friendly.
So why isn’t hemp paper used more? Well, it used to be.
The first hemp paper was made over 2,000 years ago, and before the end of the twentieth century, 75% to 90% of worldwide paper production came from hemp! But things went downhill for hemp products with the passing of the Marihuana Tax Act of 1937, and by 1970, marijuana and hemp were heavily restricted.
But today, because of the passing of the 2018 farm bill, there’s hope for a future where less sustainable practices and products are replaced with hemp products. This is good news considering there are so many benefits and uses of hemp products like hemp paper.
The Benefits and Uses Of Hemp Paper
(table of contents)
1. HEMP PRODUCES MORE YIELD AND GROWS FASTER THAN TREES
Not only does the time-saving characteristic of hemp mean that more paper can be produced at a faster rate to meet increasing global paper demands, but the higher yield production is amazing news for our forests.
With hemp requiring significantly less land to produce high yields, fewer forests and natural lands need to be cleared for paper production.
With the pulp and paper industry accounting for 40% of global industrial wood trade, it’s essential for the health of our forests and the planet, for paper production to turn over a more sustainable leaf.
2. HEMP PAPER CAN BE RECYCLED MORE THAN TREE PAPER
When it comes to the recyclability of hemp paper vs tree paper, hemp paper takes a huge lead and can be recycled more than twice as much as tree paper. That’s a huge difference!
Hemp paper can be recycled seven times, while tree paper can only be recycled three times. Considering non-recycled tree paper comes from virgin trees and is a culprit of deforestation, this is a pretty big deal.
Plus, there are loads of other benefits of recycled paper, like less water waste, fewer greenhouse gas emissions, and lower energy usage.
But that’s not all.
A lot of awesome hemp products, like hemp underwear, rely on the use of hemp’s long bast fibers, and as a result, the lesser-used hemp short fibers or hemp hurd are often left as an easily reusable waste product. While hemp paper can also be made out of long bast fibers, hemp hurd fibers can also be turned into paper products, creating a closed-loop system that saves natural resources and reduces landfill waste.
3. HEMP PAPER REQUIRES MINIMAL TO NO CHEMICAL PROCESSING
Hemp paper can be processed and bleached with environmentally friendly and non-toxic methods like oxygen delignification and autohydrolysis. These methods use more natural compounds like sodium carbonate, and non-chemical processes like the use of high pressure, temperatures, and steam.
A large part of why chemical processing is needed in paper production is to keep the cellulose (provides structure) and remove the lignin (creates unwanted brown color) found in the plant fibers.
When you compare hemp paper vs tree paper, hemp contains 55% to 77% or more cellulose while trees contain 40% to 45% cellulose. Hemp contains from 5% to 24% lignin, while trees contain 18% to 35 % lignin.
Since trees have less cellulose and more lignin, removing the lignin requires more hazardous chemical processes that can harm the environment, workers, and people who live near paper mills.
4. HEMP PAPER LASTS LONGER THAN TREE PAPER
Because of its pure, long, and strong cellulose fibers, hemp paper is known to be chemically and mechanically stable, so it’s long-lasting.
On the other hand, tree paper ends up in poor shape more quickly because of its short fibers and acidic characteristics. Tree paper is also more susceptible to environmental factors like sunlight, heat, and dampness, which can turn the paper yellow and brittle.
But rag paper (papers made of materials like hemp, cotton, and flax fibers) from the 1500s are still strong, stable in color, and flexible.
5. HEMP PAPER HELPS STOP DEFORESTATION AND SAVE THE PLANET
When comparing hemp paper vs tree paper, hemp can significantly reduce forest clear-cutting. This is in large part because one acre of hemp produces as much paper as 4.1 acres of trees, and because unlike trees, hemp can be harvested two or three times a year.
Plus, hemp has been scientifically proven to absorb more CO2 than any forest or industrial crop. So not only does growing hemp for paper fight deforestation by minimizing the clear-cutting of forests, but it helps reduce the impact of climate change by acting as the ideal carbon sink.
Forest clear-cutting damages the soil too, and even if new seeds are planted right away, the soil disturbance will lead to slower growth rates. But, after hemp is cultivated, the soil is left in optimal growing conditions.
And while clear-cutting forests lead to the destruction of animal and indigenous habitats and is a major factor in biodiversity loss, hemp has been proven to be a biodiversity-friendly crop.
6. HEMP PAPER MAKES GREAT SUSTAINABLE HYGIENE PRODUCTS
Hemp can be used to make hemp toilet paper, paper towels, tissues, tampons, diapers, and more.
Since hemp hygiene products are made with soft bast fibers, even though they are strong and durable, they’re still soft on your skin, just like hemp underwear.
7. HEMP PAPER IS AN EXCELLENT SUBSTITUTE FOR TREE PAPER FILTERS
With its composition, porosity, and resistance to water, hemp has a high filter efficiency compared to other filter materials. These properties make it an effective, sustainable alternative for all types of filters, like oil and air filters, and coffee filters.
This is great news if you’re a big coffee drinker. While most people think of the environmental impact of constantly throwing out single-use to-go coffee cups, a lot less focus is put on coffee filters you use at home or work.
But the preparation method of coffee is accountable for 30% of overall coffee emissions. Single-use tree paper coffee filters are thrown out after every use and contribute to deforestation, and the release of toxic chemicals in landfills.
Luckily, because of hemp’s natural antibacterial properties and strength, it makes a great reusable coffee filter option, putting it even further ahead of the game when you compare hemp paper vs tree paper for filters.
8. HEMP PAPER IS A BETTER OPTION FOR TEA BAGS
Similar to #7, because of its high filtration efficiency, porous fibers, and ability to maintain its structure while wet, hemp paper is a far better option for tea bags than tree paper.
Tree paper is so ineffective as a tea bag on its own that most tea bags contain some plastic to keep them together. At high temperatures, during a typical steeping process, microplastics from tea bags can be released into your cup of tea.
That’s why it’s always important to opt for tea brands that mention they are completely plastic-free, choose loose-leaf products, or opt for a more sustainable, reusable hemp tea bag instead.
9. HEMP PAPER MAKES THE PERFECT ART CANVASES
The natural characteristics of hemp fiber, including its strength, durability, and softness, make for an ideal art canvas.
Today, most canvases are made out of a combination of cotton and PVC (a synthetic non-biodegradable plastic polymer). Since you and I know that when it comes to hemp vs cotton, hemp beats cotton in sustainability and strength, and there’s no arguing the damage from depending on non-biodegradable plastic, it’s an easy win for hemp canvasses.
10. HEMP PAPER MAKES FOR RESILIENT BANK NOTES
Banknotes (or as you and I call it, cash or money) need to be stronger and more durable than other types of paper, which is why hemp’s strong fibers make it suitable for banknotes and other security papers.
Cash is often folded, crumbled, and used over and over again, which is why long lasting, strong, and durable hemp paper is a great option. Since today’s American currency consists of 75% cotton, it makes sense to switch over to the more sustainable and stronger hemp fiber.
11. HEMP PAPER MAKES FOR HIGH-QUALITY BOOKS, ARCHIVAL PAPERS, AND RELIGIOUS/SACRED TEXTS
The chemical and mechanical stability of hemp paper I mentioned in #4 makes it a great option for print items that are meant to last a while, like books, archival papers, and religious or sacred texts.
Many old tree paper books need to be handled very gently to last decades, but since hemp contains a high cellulose content and has strong and long fibers, it produces an extremely durable paper that lasts.
Archival paper is often used for publications that have significant legal or historical value, and that will be applicable for many, many years to come. Similarly, religious and sacred texts are intended to last hundreds of years, which is why it’s beneficial to print these texts on durable paper, like hemp paper.
From buying sustainable hemp underwear to ditching single-use plastic, living sustainably involves making a lot of changes, especially when it comes to products that are so widely used, like paper.
As far as hemp paper vs tree paper goes, it’s hemp paper all the way. Switching to hemp paper means producing paper that’s more durable and longer-lasting, in less time and on less land, with far more sustainable and non-toxic practices.
What paper products do you wish were made out of hemp? Share in the comments!


The MI-230 has an ultra-narrow FOV half angle of 14 degrees and the handheld unit displays and stores the current target temperature. Applications include measuring the temperatures of road surfaces, plant canopies, and soil, snow and water surfaces. Measurement of surface temperature is a crucial component of energy transfer. Accurate measurement of the leaf-to-air temperature gradient is essential to the determination of transpiration rate and stomatal conductance in both single leaves and plant canopies.
Apogee Instruments MI series infrared meters consist of a handheld meter and a dedicated infrared radiometer, which includes a thermopile detector, germanium filter, precision thermistor (for detector reference temperature measurement), and signal processing circuitry mounted in an anodised aluminium housing.
All infrared meters also come with a radiation shield designed to minimise absorbed solar radiation, but still allowing natural ventilation. The radiation shield insulates the radiometer from rapid temperature changes and keeps the temperature of the radiometer closer to the target temperature. The separate sensors are potted solid with no internal air space and are designed for continuous temperature measurement of terrestrial surfaces in indoor and outdoor environments.
The meters provide a real time temperature reading on the LCD display and come equipped with manual and automatic data logging features for making spot check measurements or monitoring temperature over time.
|Calibration Uncertainty (-20 to 65°C):||± 0.3°C, when target and detector temperature are within 20°C|
|Calibration Uncertainty (-40 to 80°C):||± 0.5°C, when target and detector temperature are different by more than 20°C|
|Measurement Repeatability:||± 0.1°C|
|Stability (Long-term Drift):||less than 2% change in slope per year when germanium filter is maintained in a clean condition|
|Field of View:||14° half angle|
|Spectral Range:||8 to 14 µm; atmospheric window|
|Operating Environment:||0 to 50°C, < 90 % non-condensing relative humidity up to 30°C, < 70% non-condensing relative humidity from 30°C|
|Meter Dimensions:||12.6 cm length, 7.0 cm width, 2.4 cm height|
|Sensor Dimensions:||2.3 cm diameter and 6.0 cm length|
|Mass:||270g (with radiation shield)|
|Cable:||2m of four conductor, shielded, twisted-pair wire. Additional cable available. Santoprene rubber jacket (high water resistance, high UV stability, flexibility in cold conditions)|


We had our first cooking club today and it was terrific! Thank you Mrs. Wheaton and Mrs. Cistone for leading it! We got to mix together oats, peanut butter, chia seeds, honey, dark chocolate chips, and pumpkin to make a yummy concoction. Then the fun began as we got to mix it with our hands and roll it into balls. We learned about measuring 1 cup, 1/2 cup, and 1/4 cup. We learned why chia seeds are healthy for us. We learned how not to spread germs when we are cooking. And as one kiddo said...we learned patience! When we were done making the balls, we then got to make banana ghosts. Of course I wanted to eat them all! What a fun morning!


Crescent Moon + Rutilated Quartz
Sorry, currently out of stock
Crescent Moon + Rutilated Quartz wrapped in blackened copper. Including bail, measures 3.25". Finished off with a long, 28" solid copper chain. If you would like a different length, please leave me a note at checkout.
Rutilated Quartz is a transparent Quartz with Rutile inclusions that are in hairlike growths. Making each one completely unique.
▲ Metaphysical properties: Quartz has been utilized for millennia as a potent worker of magic. It is an extremely vibrational stone that promotes both balance, and healing.
▲ Copper is a great conductor of energy. It is one of the oldest metals in use and has been widely associated with ancient Alchemy and Astrology. Representing the Goddess Aphrodite and Venus, it is a metal that lends itself to beauty, balance, clarity, and creativity. It is particularly useful in strengthening bonds between people.
The blackened copper is hand oxidized. To ensure the integrity of the patina, please do not shower or submerse your piece in water.
Completely handmade by nature and me. Comes wrapped and packaged in a gift box.