Replace Toxic Products in your home

Everyday Roots

This book includes home remedies, natural beauty recipes and Diy household product tutorials. Discover over 215 suprising natural home remedies using common ingredients like onion, lemons and apple cider vinegar. EveryDay Roots will help you to make healthy changes in your life. Learn how to treat coughs, headaches and other health conditions with common ingredients like honey and watermelon. When you buy the book you get a 328 page Pdf with a clickable table of contents. More here...

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Author: Claire Goodall
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Detection Of Tissue Transglutaminase Activation By Cell Resistance To Detergents

This protocol is a simple and rapid approach to identify apoptotic cells with activated TGase 2. The method is based on the propensity of crosslinked protein to withstand treatment with detergents. The authors have noticed that when live, nonapoptotic cells are subjected to treatment with solutions of nonionic detergents, lysis of their plasma membrane and release of the content of cytoplasm is complete, resulting in preparation of isolated nuclei. In contrast, apoptotic cells resist the detergent treatment their cyto-plasmic protein remains insoluble, attached to the nucleus in the form of a shell-like cover (Grabarek et al., 2002). It is possible, therefore, by flow or laser scanning cytometry to distinguish apoptotic cells from the nuclei isolated from nonapoptotic cells, by means of the abundance of protein in the former. In addition, bivariate gating analysis of cellular DNA and protein content makes it possible to reveal the cell cycle distribution separately for the population...

Household Chemicals as Urine Adulterants

Simple household chemicals are found to be effective adulterants of urine drug tests. These include table salt, vinegar, liquid laundry bleach, concentrated lemon juice, and Visine eye drops (8,9). The effectiveness of these chemicals on specific drug tests is summarized below. In addition, Uebel and Wium (12) also studied the effect of common household chemicals such as Jik (a South African brand of bleach sodium hypochlorite), Dettol (Reckitt Benckiser UK chloroxylenol), G-cide Plus (JAST International glutaraldehyde), Pearle Hand Soap, ethanol, isopropanol, and peroxide in causing false-negative results when used as adulterants in urine specimens. Most of these chemicals interfered with toxicological screening results using EMIT DOA urine-test reagents. Glutaraldehyde and Pearle Hand Soap had the greatest effect (false negative) on a methaqualone test. Chlorox-ylenol and Pearl Hand Soap also demonstrated maximum effect in causing a false negative in cannabis tests.

Laundry Detergents

Another important class of compounds produced by biotechnology is enzymes. These protein catalysts are used widely in both medical and industrial research. Proteases, enzymes that break down proteins, are particularly important in detergents, in tanning hides, in food processing, and in the chemical industry. One of the most significant commercial enzymes of this type is subtilisin, which is produced by a bacterium. Because many stains contain proteins, the manufacturers of laundry detergents include subtilisin in their product. Subtilisin is 274 amino acids long, and one of these, the methionine at position 222, lies right beside the active site of the enzyme. This is the site on the enzyme's surface where the substrate is bound, and where the reaction that is catalyzed by the enzyme takes place. In this instance the substrate is a protein in a stain, and the reaction results in the breaking of a peptide bond in the backbone of the protein. Unfortunately, methionine is an amino acid...

Virion and Genome Structure and Composition

Based on gel analyses, ascovirus virions contain at least 12 structural polypeptides, ranging in size from 12 to 200 kDa. In addition to proteins and the DNA genome, the presence of an envelope, as detected by electron microscopy as well as experiments with detergents and organic solvents, indicates that virions contain a substantial lipid component. And, as in other enveloped viruses of eucaryotes, it is likely that the virion also contains carbohydrate in the form of glycoproteins, though none have yet been identified.

Recent Studies on the Possible Identity of Microsomal CATa

Although CATA can be solubilized from rat liver microsomes using mild detergents such as deoxycholate5 and reconstituted into liposomes of known phospholipid compo-sition10 without loss of activity, further purification of the enzyme away from other membrane components requires harsher conditions, which inactivate the enzyme. For example, substantial further purification can be achieved by extraction into 8mM CHAPS with 4 M urea followed by anion-exchange chromatography and gel filtration (N. M. Broadway & E. D. Saggerson, unpublished work) but the protein then has to be detected by indirect means. Because of these difficulties, purification of microsomal CATa to homogeneity has not yet been achieved. It was during attempts to purify CATa, prior to sequencing, that we arrived at the notion that microsomal CATa and mitochondrial CFT, may be very similar proteins.

Solubilization and Purification of CFTR Protein Using Pentadecafluorooctanoic Acid PFO

Purification of membrane proteins requires their dissociation from interacting cellular proteins and membrane phospholipids using compatible detergents. The membrane protein of interest can then be purified from the mixed protein-detergent micelles by virtue of distinctive physicochemical properties or specific affinity with an immobilized ligand. CFTR is a very hydrophobic membrane protein and is poorly soluble in most detergents except the strong ionic detergent sodium dodecyl sulfate (SDS) and the salts of PFO (1,20). In the original method we published with J. R. Riordan, CFTR solubilized in SDS was purified primarily by affinity to hydroxyapa-tite. Unfortunately, this purification procedure is quite complex and time-consuming (1). Several membrane proteins have been purified by an easier method, using nickel-affinity chromatography. This method relies on the affinity of a polyhistidine tail (His), engineered onto the amino or carboxyl terminus of the protein, to nickle resin...

Virion Structure and Composition

As indicated by the size and complexity of the virions, the genome of ascoviruses is large, and consists of a single molecule ofdouble-stranded circular DNA. The genomes of three species have been sequenced, the type species SfAV, TnAV, and HvAV. The SfAV genome is 157 kbp and codes for at least 120 proteins (Figure 3), whereas the TnAV 2a genome is slightly larger, 174 kbp, and codes for at least 134 proteins. The genome of HvAV is 186 kbp and codes for approximately 180 potential proteins. Based on gel analyses, ascovirus virions contain at least 12 structural polypeptides ranging in size from 12 to 200 kDa. In addition to proteins and the DNA genome, the presence of an envelope as detected by electron microscopy, as well as experiments with detergents and organic solvents, indicate that virions contain a substantial lipid component. And, as in other enveloped viruses of eukaryotes, it is likely that the virion also contains carbohydrate in the form of glycoproteins, though none...

Epidemiology And Prevention

Patients contribute to disseminate C. difficile toxigenic strains in hospital settings where endemic and epidemic situations may occur. To prevent cross-contamination of patients with C. difficile, contact precautions must be implemented until diarrhea stops. However, C. difficile spores can survive for months in hospital environments, and patients can be contaminated via contact with care personnel or contaminated objects. Moreover, spores can resist several commercial detergents commonly used to clean surfaces in healthcare institutions. Chlorine-based disinfectants appear to be a better choice and may contribute to reduce the incidence of C. difficile infections. 15 However, to limit C. difficile-associated diarrhea in hospitals, measures must include a combination of rational use of antibiotics and better infection control. 16

Bcl2 Family Of Proteins

Bax and Bak exist as monomers in aqueous solution but can form homo-oligomers in the presence of detergents. These large homo-oligomers are thought to form channels with a pore size large enough to allow passage of proteins such as cytochrome c, though direct biochemical and structural evidence

Direct Nonhealth Care Costs

Fecal incontinence compared with other disorders does not appear to have a heavy overall impact on the NHS. Indeed, as already pointed out, the costs related to the disorder are almost entirely borne by the patients themselves or their families. In addition to the cost of pads, other costs have to be borne entirely by the patient. The above-mentioned Dutch study 5 estimated expenditure for antidiarrheal drugs (used by 26 of the patients interviewed), skin care products (11 ), special articles of clothing (10 ), cleaning products (9 ), and special foods (6 ).

Lysis Buffers And Other Solutions

Ature. 44,55,56 The various formulations for ''lysis buffers'' generally consist of a Tris buffer at a pH between 7.5 and 8.0, EDTA (chelator), and sodium dodecyl sulfate or n-lauroylsarcosine (detergents to lysis cells). These buffers have demonstrated to be effective at room temperature in the preservation of DNA in blood samples for 6 months 44 and tissue samples for 2 years or more. 39 The use of a dimethyl sulfoxyde (DMSO) salt solution for the preservation of DNA in various tissues has been demonstrated to be effective for storage from 6 months to over 2 years. 39,44,48 This solution of 20 DMSO, 0.25 M EDTA, and NaCl 5 to saturation, pH 7.5 44,48 was found to be the most effective method of noncryogenic storage for the prevention of DNA degradation. 39

Patents and the Rise of Biotechnology Companies

Biotechnology has also been sucessful in development of other useful products. Today many laundry detergents contain proteases, enzymes that remove stains by digesting the protein components of the stain. However, such enzymes are inactivated by bleach. In 1988 the biotechnology company Genecor received approval for a bleach-resistant protease. This had been accomplished by isolating the gene for protease and then, using site-directed mutagenesis, changing the gene such that the corresponding protein was no longer sensitive to inactivation by bleach.

Structure And Channel Forming Activity Of The Bcl2 Proteins

Channels formed by oligomeric Bax have multiconductance levels, ranging from a few pS up to 4-5 nS, are pH-sensitive, slightly cation-selective, and Ca2+-insensitive (19,20). Monomeric Bax does not possess channel-forming activity. Oligomerization of Bax can be induced artificially by several detergents (22). Recombinant Bax or cytosolic Bax exposed to Triton X-100 or octyl gluco-side form oligomers with a molecular weight of 80 kDa (23). These oligomers are able to form at least dimers resulting in oligomers of 160 kDa. This would correspond to Bax tetramers and octamers. Oligomerization only appears to take place when the detergent is present over the critical micelle concentration. This is supported by the nuclear magnetic resonance (NMR) structure study where a

Physical Properties

Like other enveloped viruses, hantaviruses are sensitive to detergents and organic solvents. Other physical properties of virions include sedimentation densities in sucrose of 1.16-1.17 g ml-1 and in cesium chloride of 1.20-1.21 g ml-1. Disruption of virion particles with nonionic detergent releases the nucleocapsids, which sediment at densities of approximately 1.18 g ml-1 in sucrose and 1.25 g ml-1 in cesium chloride. HTNV infectivity is generally stable for up to 12 h at 0 4 C. Nonphysiological pH extremes (< 7.0 or

Environmental Toxicity

Environmental safety of spent microbicides is an issue that is not widely acknowledged because the use of such chemicals is deemed to benefit humans. However, now that microbicides are incorporated into many consumer products, both human and environmental exposures to these chemicals have already increased considerably. Most microbicides also contain detergents and inert ingredients that may potentiate their action and could also exacerbate their toxicity. There is little direct evidence of this because it has not been widely or systematically examined, but increasingly we are beginning to understand that complex chemical mixtures pose particularly significant environmental risks even when individual chemicals may be below their acknowledged toxicity threshold 4 . Unfortunately, toxicity of the active ingredients of microbicides and their by-products is poorly investigated under relevant conditions.

Materials Compatibility

A desirable attribute of microbicides for use in healthcare is materials compatibility. For medical instruments and many pipes and surfaces that are disinfected regularly, the chemical concentrations needed to ensure effective microbicidal action can also cause corrosion or damage. Microbi-cides are therefore often formulated to enhance their materials compatibility. For example, acidic detergents used as sanitizers usually contain phosphoric acid, which is less corrosive than, say, hydrochloric acid, even though a sanitizer containing hydrochloric acid may be a more effective microbicide 52 . A second example is rubber that is continuously or intermittently exposed to reactive chemicals the rubber components used in milking equipment are regularly washed with chlorine solutions, which tend to ''crack'' or craze the surfaces 53 . Such damage increases with exposure and if these items are not replaced regularly, the milk can contain small rubber carbon particulates, causing it to be...

Innovative Products and Technologies

In general, the trend in microbicide innovation is to steer away from products that have significant recognized toxic residuals. This trend is less prominent in North America than in Europe where imminent regulation 67 will limit the types of chemicals used and discharged in many areas. Even in North America there is increasing recognition that it may be possible to use environmentally friendly microbicides. When this is combined with the well-recognized mantra that cleaning is an absolute prerequisite for effective disinfection of objects and surfaces, the trend toward oxidative products that are themselves relatively good cleaners is fairly obvious. There is a real upward, or one could say backward, swing to peroxygen compounds. Hydrogen peroxide is both a naturally occurring chemical and among the earliest recognized disinfectants. While peracids were not recognized early on, they are undoubtedly naturally generated in certain situations. Nowadays, peracids, especially peracetic...

Main Destructive Influences On

Under the influence of UV light (including sunlight) and acids, DNA contained in biological stains as well as extracted DNA breaks into pieces (degrades). Depending on the intensity of fragmentation, PCR might still be possible. Humidity does not directly affect DNA but will allow mold and bacteria to destroy the sample including the DNA within days. Frequent freezing and unfreezing of stains or extracted DNA will also lead to degradation. Household use of detergents and cleaners does not necessarily destroy DNA. 19 Sperm heads on fabric can survive machine washing at 30-40 C if no bleach was used.

External Disinfection of Dialysis Monitors

Materials of which the body of the machine and relative accessories are composed to avoid irreparable damage which could range from the simple opacity of the transparent panels to the actual 'melting' of some of the plastic components. On the whole, products containing benzene, acetone, toluene, xylene or similar solvents should be avoided. Finally external detergents and disinfectants should be used with disposable paper wipes (to be changed after use on each machine) avoiding every type of cross-contamination. Other critical components, possible causes of transmission of bacteria and or viral cross-infections from patient to patient, are constituted by the connection of the dialytic solution in the direction of and from the filter. Both the external and internal surfaces can be easily contaminated especially in the dismantling phase of the dialyzers, when the maneuver is carried out by operators wearing soiled gloves. Moreover, at the beginning of the dialytic session, before...

Properties of the Virion

HIV particles have a diameter of 100-120 nm and a spherical morphology classical of several other retroviruses. They consist of a dense truncated cone-shaped core surrounded by a membrane bilayer (envelope) decorated with spikes corresponding to the surface glycoprotein (SU). HIV particles are assembled beneath the cell surface. The internal structure of the virus particle rearranges after budding, a result of the proteolytic cleavage of the major structural components by a viral-encoded protease. This process is called maturation. Virions are sensitive to radiation, acidic pH, diluted ethanol, detergents, chorine and temperature (60 C). The

Selected Examples Of Amino Acids

For these considerations and their possible consequences in understanding the physiological and biochemical background of glutamate excretion, a critical evaluation of the methods used for inducing glutamate production by C. glutamicum is necessary (79). Several different kinds of treatments lead to glutamate excretion. Based on the original observations, glutamate was found to be excreted under conditions of biotin limitation (2). Corynebacterium glutamicum requires biotin for growth. Biotin is a cofactor of several enzymes, most importantly of pyruvate carboxylase and acetyl-CoA carboxylase, which is essential for fatty acid synthesis. Glutamate overproduction can also be induced by addition of some kind of detergents, e.g., polyoxyethylene sorbitane monopalmitate (Tween 40) or polyoxyethylene sorbitane monostearate (Tween 60). The reason for induction of glutamate excretion is definitely not simply an increase in membrane permeability, because closely related detergents like the...

Sample Preparation and Matrix Effects

Although solvent extraction methods might be expected to produce a sample more amenable to immunoassay than the digestion method, these methods can present serious challenges. A solvent extract of a hair sample will not contain keratin, for example, but it will contain a significant and likely variable amount of lipid. When the solvent is evaporated, which it must be because only a small amount is tolerated in any immunoassay, the lipid is left to be partially solubilized or suspended in an aqueous medium added to the dried extracts. Detergents can be added to the extract to aid in the solubilization of the lipid, but this must be carefully monitored and controlled to avoid damaging the antibodies or enzymes in the subsequent immunoassay. Too much detergent can affect primary or secondary antibody binding, or can cause detachment of antibody bound to solid phase such as in microtiter wells. Variations in amounts of lipid among different hair samples, and in micelle formation when...

Physical Properties of the Virion

JE virus infectivity is readily lost by heat, acid, lipid solvents and detergents. Lipid solvents (ether and chloroform), or ionic detergents (sodium deoxy-cholate and sodium dodecyl sulfate) inactivate both infectivity and hemagglutinating activity (HA). Although nonionic detergents (Triton X100, or Nonidet P40) destroy virion structure and infectivity, HA is retained.

In Situ Generation of Active Species

The laundry detergents industry has long relied on the in situ generation of peracetic acid, but applications have been largely focused on bleaching. The main peracetic acid precursors in use today are tetraacetyl glycol urea, glucose pentaacetate, diacetyldioxohydrotriazine, sodium nonanoyloxyben-zenesulfonate, and TAED. TAED was first used by Lever in France in 1978

Midgut Secretory Mechanisms

Like all animal proteins, digestive enzymes are synthesized in the rough endoplasmic reticulum, processed in the Golgi complex, and packed into secretory vesicles (Fig. 4). There are several mechanisms by which the contents of the secretory vesicles are freed in the midgut lumen. During exocytic secretion, secretory vesicles fuse with the midgut cell apical membrane, emptying their contents without any loss of cytoplasm (Fig. 4A). In contrast, apocrine secretion involves the loss of at least 10 of the apical cytoplasm following the release of secretory vesicles (Fig. 4B). These have previously undergone fusions originating larger vesicles that after release eventually free their contents by solubilization (Fig. 4B). When the loss of cytoplasm is very small, the secretory mechanism is called microapocrine. Microaprocrine secretion consists of releasing budding double-membrane vesicles (Fig. 4C) or, at least in insect midguts, pinched-off vesicles that may contain a single or several...

Architecture of the NS2 protease domain

NS2 is a hydrophobic protein with several putative transmembrane segments. Removal of the N-terminal region containing these segments greatly increased the expression and yield of recombinant NS2 without affecting auto-proteolysis.31'42 The C-terminal domain of NS2 residues 94 to 217 (NS2pro) was expressed in bacteria and purified to homogeneity in the presence of detergents. The crystal structure revealed that NS2pro consists of two subdomains connected by an extended linker (Fig. 5A). The N-terminal subdomain begins with two antiparallel a helices (H1 and H2) connected by a short loop. Following the second helix, the subdomain has a random-coil conformation that contacts both H1 and H2. The protein then continues into a long, extended coil before entering the antiparallel p sheet in the second subdomain. The last p strand (b5) continues to the C terminus of NS2.

Combination Dnarna Fish and Immunophenotyping

In the last few years, several protocols have been developed allowing simultaneous detection of DNA or RNA sequences together with proteins. Before applying one of these protocols it is important to consider that DNA and RNA molecules differ in chemical nature and in localization within a cell. The consequence of this is that DNA and RNA FISH have different requirements concerning pretreatment of cells and hybridization conditions. Hybridization to DNA sequences requires denaturation of the target DNA, whereas RNA sequences are single-stranded. However, owing to the high structural complexity of many mRNAs, a controlled denaturation of target sequences often improves RNA hybridization signals (Dirks et al., 1993). The genomic DNA is tightly packed within the cell nucleus, so detection of a specific DNA sequence present somewhere within this genome requires measures to obtain full accessibility of the nucleus and removal of cellular components that may cause background signals....

Discovery and Classification

Investigation of bacteriophages of phytopathogenic pseudomonads led to the discovery of a virus (cj> 6) which was extremely sensitive to organic solvents and detergents. The first reports in 1973 described two unique features this spherical virus had a lipid envelope and the genome was composed of double-stranded RNA (dsRNA). Phage < j)6 was, until very recently, the only member of the family Cystoviridae, genus Cystovirus. It appears now that 6-like viruses can be isolated from sources such as degrading plant material. The host range of this virulent phage is restricted to pseudomonads. The largest number of sensitive strains is found among glysinea, phaseoli-cola, syringae, tabaci and viridiflava species.

The Postsynaptic Density

If synaptosomes are exposed to strong detergents such as Triton X the pre- and postsynaptic membranes are released from the rest of the complex (Figure 17.2). This, the so-called synaptic plasma membrane (SPM), may then be isolated from the other elements of the synaptosome by density gradient centrifugation. The fact that the pre- and postsynaptic membranes stay together when the rest of the synaptosome has been liquidated suggests that they are held together by some electron-translucent matrix. The analysis of this region can be carried a step further by subjecting the SPMs to 3.9 w v sodium lauryl

Biophysical Properties

The surface of cricket paralysis virus (CrPV) does not show the characteristic deep canyon around the fivefold axes - which in the case of the former is where the receptor-binding site is known to be. The mature virions have a buoyant density in neutral CsCl ofbetween 1.34 and 1.39 g cm-3 and sedimentation coefficients that range between 153S and 167S. For those viruses where physico-chemical stability has been assessed, for example, CrPV, the virions are stable at pH 3.0 and are resistant to treatment with detergents and organic solvents such as ether and chloroform.

Overview Of The Enterohepatic Circulation Of Bile Acids

Bile acids are amphipathic physiological detergents that play essential roles in promoting absorption, excretion, and transport of cholesterol, lipids, lipophilic nutrients, and other hydrophobic compounds in the liver and the intestine.1 The two primary bile acids in humans are cholic acid (CA) and chenodeoxycholic acid (CDCA). In the intestinal bacterial flora these can be converted to secondary bile acids, deoxycholic acid (DCA) and lithocholic acid (LCA), respectively.

Control Of Bile Acid Transport And Metabolism

In addition to their role as physiological detergents, bile acids possess crucial regulatory properties which allow them to control their own transport and metabolism within the enterohepatic circulation through multiple feedforward and feedback mechanisms. Hepatocytes and enterocytes possess numerous signaling pathways that are activated or modulated by bile acids, and ultimately serve to maintain intracellular concentrations of potentially toxic bile acids at a constant level.

A Poly Acrylamide Gel Electrophoresis PAGE

While IEF is a powerful tool used primarily for confirming the identity of a protein biopharmaceutical, upon suitable validation it can also be used as a stability-indicating method to monitor changes to the protein over time 32, 33 , Isoelectric focusing is normally run in a native gel using wide pore polyacrylamide or agarose but the addition of nonionic detergents, nonionic chaotropic agents or reducing agents during sample preparation may serve to dissociate molecular complexes and aggregates resulting in enhanced resolution.

Signaling Events Measured by Protein Phosphorylation

Signaling events in single cells may also be monitored by flow cytometry by using intracellular phospho-protein staining techniques. Measurement of protein phosphorylation with phospho-specific antibodies has given insight into kinase signaling cascades within cells after stimulation. Several groups have demonstrated staining of phospho-epitopes for flow cytometric analysis (11). Among the molecules previously examined are Statl (12,13), Stat4 (14), Akt (15,16), ERK and MEK (17), cJun and p38 (18), and various others (19). The methods used to prepare cells for staining with phospho-specific antibodies differed in each case, but they generally employed a fixation step with formaldehyde, followed by permeabilization with alcohols, detergents, or saponin. Because many of the epitopes to be recognized are novel and might be sequestered in protected locales within cells, it remains unclear whether there exists a general method (or set of methods) by which most phospho-epitopes can be...

Polymerase Activity

A viral RNA-dependent RNA polymerase is activated after treatment of LNYV and BNYV cytorhabdovirus virions with mild nonionic detergents. This activity cosediments with the 40 45S loosely coiled nucleocapsid filaments that are released from virions by detergent treatment. The transcribed products are complementary to the genome, as expected of mRNAs. Thus, the described polymerases of these plant rhabdoviruses appear to be similar to the extensively studied polymerase of the animal rhabdo-virus prototype, VSV.

Sample Preparation for PFGE

Analysis of fragile, high-molecular weight DNA molecules that easily break into small pieces. To prevent DNA damage, intact cells mixed with warmed, liquid agarose are pipetted into plug molds, 10 x 5 x 1.5 mm in size. Cells embedded in the agarose plugs are lysed in situ by detergents and enzymes. The agarose matrix keeps the large DNA molecules intact, while permitting diffusion of lysed cellular components from the plug. Infrequent-cutting restriction endonucleases are used to digest the DNA. The choice of restriction enzyme is critical because it should generate between 10 and 30 numbers of DNA fragments necessary for strain discrimination. A list of endonucleases for typing specific microorganisms have been published. 9 After restriction enzyme digestion, the plugs are cut into appropriate sizes based on the DNA concentration and loaded into the wells of an agarose gel. The gel is then placed in the electrophoresis chamber, and pulsed field conditions are chosen based on the...

Structure and Replication

The virions contain a lipid envelope, have a buoyant density of 1.19-1.3 gml-1, and are sensitive to inactivation by heat, acid pH, organic solvents, and detergents. Currently the virion structure is best understood for dengue viruses and TBEVs. Dengue viruses, as reconstructed from cryoelectron microscopy (cryo-EM) images at 24 A resolution, exist as a smooth particle comprised of dimeric units of the envelope (E) protein arranged in a head-to-tail fashion on its surface with icosahedral symmetry, encasing a nucleocapsid composed of capsid protein and genomic RNA (Figure 1). These genomes are on the order of 11 kbp (42 S sedimentation) and differ from other flavivirus genera in having a 5' cap. The 5' untranslated region (UTR) encodes a unique secondary RNA structure. The 3' UTR contains conserved sequences and secondary structures, including a unique terminal stem-loop structure required for viral RNA replication (Figure 2). Such structures are found among both mosquito and...

Severe Acute Respiratory Syndrome

Cleaning and disinfection of environmental surfaces are important components of preventing nosocomial infections in healthcare settings. Although little is known about the extent of environmental contamination in SARS patients' rooms, epidemiologic and laboratory evidence suggests that the environment could play a role in transmission 113 . Therefore, cleaning and disinfection are critical to the control of SARS-CoV transmission too. Environmental cleaning and disinfection for SARS-CoV follow the same principles generally used in healthcare settings. CDC recommends using EPA-registered hospital detergents or disinfectants and following manufacturer's recommendations for use-dilution (concentration), contact time, and care in handling. Special attention should be given to frequently touched surfaces and horizontal surfaces around the patient after aerosolgenerating procedures 116 .

Capsid Organization and Stabilizing Interactions

The sobemovirus capsid is constructed from a protein of about 30kDa. A proportion of coat protein subunits in SBMV and TRoSV virions exist as stable dimers that are preferentially linked to RNA. The basic SBMV capsid structure is T 3 quasi-symmetry of 180 subunits. High resolution x-ray diffraction has revealed three types of quasi-equivalent subunits, namely A, B and C, which are chemically identical but have different conformations. The A subunits cluster at the fivefold axes, whereas sets of B and C aggregate at the quasi-sixfold vertices. Each subunit consists of a random domain, i.e. the N-terminal 'arm' located towards the virion interior, and the surface domain, which is organized into an eight-stranded antiparallel ft barrel and five a helices. Considerable similarity exists between the organization of SBMV capsid with those of tomato bushy stunt, tobacco necrosis, poliovirus type 1 and human rhinoviruses. Sobemovirus virions are extremely stable in vitro in their native...

Endogenous and Exogenous Contamination and Other Factors

Effect on PCR when irradiated with UV light (75). The inhibition is dependent on the UV dose. Unirradiated mineral oil has been reported as a facilitator in oil-free reactions containing high concentrations of nonionic detergents (76). This author suggested that components of detergent preparations (monomers, micelles, or impurities) could be responsible for adverse effects on the specificity of annealing. It is likely that detergents allow the greater solubilization of inhibitors that might otherwise aggregate and precipitate during preparation or in the reaction tube. Oil overlays may facilitate amplification by segregating inhibitors at the oil-water interface and remain an option in thermal cyclers designed for oil-free reactions.

Products derived from genetically engineered microorganisms

Appropriate cloning vector, and inserting it into a host cell such as E. coli or Saccharomyces cerevisiae. The initial application of this technology was in the micro-bial production of medically important proteins such as insulin and epidermal growth factor (Table 17.6), however other proteins may also be produced by these means. These include enzymes used in diagnostic and analytical applications, where a higher purity of preparation is required than, for example, the enzymes used in detergents. These are often derived originally from other microorganisms for example the thermostable DNA polymerase from Thermus aquaticus used in PCR is now commonly made by recombinant E. coli cells that have been transformed with the T. aquaticus gene. Many of the more recent recombinant human proteins to be developed for therapeutic use have been too complex for expression in a microbial system (e.g. Factor VIII), so it has been necessary to employ cultured mammalian cells.

Preliminary Operations

The room and the largest containers are cleaned with detergents or acids, the manipulations being done with non-powdered plastic gloves only. The containers are rinsed with distilled water, then with ultrapure water (10 successive baths) and dried. The flasks and tubes which will be directly in contact with the samples are washed in an ultrasonic bath with diluted detergents, then rinsed in 10 successive baths of tap water and 10 successive baths of ultrapure water, dried in an oven and stored in aluminum foil or in closed containers. All the instruments are decontaminated in the ultrasonic bath, using the same protocol.

Viral Membrane Proteins

The proteins of viral membranes, like those of other membranes, may be classified as either integral or peripheral. Integral proteins are those that span the membrane one or more times, and thus cannot be solubilized without disrupting the bilayer, e.g. with detergents. Peripheral proteins do not cross the membrane, and can be removed from it and solubilized by treatment with aqueous salts or chaotropic agents, which do not destroy the bilayer. Integral membrane proteins of enveloped viruses generally span the membrane only once an exception is the El protein of coronavirus, which spans the membrane three times. Each membrane-spanning, or transmembrane, or anchoring, domain is a sequence of 18-27 predominantly hydrophobic amino acid residues. Transmembrane sequences are inherently insoluble in water, so that integral membrane proteins require the presence of detergents to be soluble. In the absence of detergents or lipids, membrane proteins tend to aggregate as rosettes, with the...

Substrate Specificity And Transport Mechanism Of The

Mammalian CRTs from mouse, rat, rabbit, cow and man contain 635 amino acids and are 96 identical at the amino acid level. Each is predicted to contain 12 transmembrane spanning domains (TMs) with the amino- and carboxy-terminal regions facing the cytoplasmic side of the membrane. Two potential sites for N-linked glyco-sylation are found in the extracellular loop between TM3 and TM4. There is another predicted glycosylation site located in the extracellular loop between TMs 11 and 12 (Snow and Murphy, 2001 Sora et al., 1994). However, this is a very short loop and it may not be accessible for glycosylation. The lectin, wheat germ agglutinin (specificity for N-acetylglucosamine), was used in the purification of bovine CRT from membranes of HEK293 cells expressing high levels of the transporter (West et al., 2005). N-glycosidase F digestion reduced the mass for the purified CRT (70-80 kDa) to 50kDa. The mass for deglycosylated CRT is less than predicted from the amino acid sequence...

Optimized Fixation and Immunofluorescence Staining Methods for Dictyostelium Cells

Draw The Setup For Fixation

Recent years have seen a powerful revival of fluorescence microscopy techniques, both to observe live cells and fixed objects. The limits of sensitivity, simultaneous detection of multiple chromophores, and spatial resolution have all been pushed to the extreme. Therefore, it is essential to improve in parallel the quality of the structural and antigenic preservation during fixation and immunostaining. Chemical fixations are broadly used but often lead to antigenicity loss and severe membrane damages, such as organelle vesiculation. They also must be followed by membrane permeabilization by detergents or solvents, which can lead to extensive extraction and cytosol leakage. Fixation with solvents bypasses the need for permeabilization, but when carried out at high temperatures, leads to severe extraction of soluble proteins and lipids and cytosol wash-out, and has therefore been used routinely to visualize the cytoskeleton. Here, we describe a few modifications to the common aldehyde...

Bioreactor Configurations 341 Submerged Fermentor Systems

Airlift Submerged Bioreactor

The advantages of using a hollow fiber reactor for microbial systems include high density of cell growth, using a perfusion system for simultaneous separation of product and biomass, and biocatalyst regeneration. However, a major disadvantage is the difficulty in monitoring and controlling the growth and metabolism of the culture. Other process constraints associated with microbial hollow fiber reactors are low oxygen transfer rates at high cell density and blockage, and rupture of the membranes due to excessive growth. The accumulation of toxic products in the hollow fiber might also inhibit the metabolic activity of the cell system. Further, the effect of microbial containment on physiology, long term viability, and productivity remains unclear. The technique has been used in the production of lactic acid (35), conversion of l-histidine and biosynthesis of -galactosidase.

Reconstitution of CFTR Protein into Liposomes from PFO

To study the structure and function of a purified membrane protein, it is essential that the detergent in which the protein is purified be exchanged for phospholipids. This exchange is a time- and concentration-dependent process. Typically, phospholipids are added to detergent-solubilized proteins and then the detergent is removed by dialysis or detergent adsorption resins (24,25). Only when the detergent concentration decreases below a critical value (critical micelle concentration or CMC) can detergent monomers dissociate from the micelles and proteoliposomes be formed (26). Detergents with high CMC values, such as PFO, are easily removed by the dialysis method (20,27).

Water And Wastewater Disinfection Treatment

A wide range of nonoxidizing organic and inorganic chemicals are used for, or are able to provide, disinfecting effects, including aldehydes (formaldehyde and glutaraldehyde), phenolics, alchohols (ethanol and isoproponal), cationic detergents, nitrites, and heavy metals (e.g., mercury, silver nitrate, tin, arsenic, copper). Although most of these chemicals have little relevance for the disinfection of waters, wastewaters, or sludges, there are two noteworthy exceptions. Specifically, silver-impregnated filters are sometimes marketed for point-of-use water conditioning devices, such as those that are sometimes screwed onto the outlets of sink faucets. In this instance, the silver is intended to be slowly leached from the filter medium (typically, activated carbon) at a rate that, hopefully, will retard the opportunistic formation of microbial biofilms intent on using sorbed organics as their energy source. A second nonoxidizing chemical disinfectant option is that of using cationic...

Substrate Specificity Of Pglycoprotein And Nature Of The Drugbinding Site

Pgp displays a remarkable ability to interact with, and transport, a large variety of compounds, ranging from chemotherapeutic drugs to peptides. Most preferred substrates are amphipathic and relatively hydrophobic, although some are not (colchicine, for example, is quite water soluble). Pgp substrates range in size from large complex molecules such as paclitaxel and vinblastine to smaller drugs such as daunorubicin and doxorubicin. Pgp also interacts with linear and cyclic peptides and ionophores, including gramicidin D, valinomycin, N-acetyl-leucyl-leucyl-norleucinal (ALLN), leupeptin, pepstatin A, and several bioactive peptides.75'108 Small tripeptides such as NAc-LLY-amide are excellent transport substrates.26 Even nonionic detergents such as Triton X-100 and nonylphenol ethoxylates interact with pgp.91'109'110 Many substrates, but not all, contain planar aromatic rings and positively charged tertiary N atoms. No highly conserved recognition elements have been found in Pgp...

Metabolism and Excretion of Toxicants

Leupeptin Plant

One of the unique facets of toxicant metabolism is that even though structures of these potentially toxic products, be they natural or synthetic, are so tremendously varied, the body seems to have evolved detoxifying processes that can cope with almost any of the many different compounds. Animals possess enzymes that can metabolize drugs, pesticides, secondary plant metabolites, and synthetic compounds as defense mechanisms, which are likely because of evolution in response to selective pressures for protection against many naturally occurring toxic products. There are two categories of animal enzyme systems (1) those for the transformation of normal endogenous chemicals in tissue, such as nutrients and metabolic by-products of nutrients and (2) those that alter structure of many foreign compounds and essentially have no established normal endogenous substrates. The first category of enzyme systems has been studied in detail for their general biochemistry. These are enzymes of...

Nonylphenol Ethoxylates PEGn Nonyl Phenyl Ether Polyoxyethylene n Nonyl Phenyl Ether

Their general formula is C9H19C6H4(OCH2CH2)nOH. Each non-oxynol is characterized by the number (n) of ethylene oxide units repeated in the chain for example, nonoxynol-9, nonoxynol-14. They are present in detergents, liquid soaps, emulsifiers for creams, fabric softeners, photographic paper additives, hair dyes, lubricating oils, spermicides, and anti-infective agents. They are irritants and sensitizers. Nonoxynol-6 was reported as a sensitizing agent in an industrial hand cleanser and in a crack-indicating fluid in the metal industry. Nonoxynol-9 is the most common

The Alternative Pathway of Complement a Pattern Recognition System

Higher vertebrates have established effective and highly sophisticated immune response in order to combat foreign microbes and to maintain tissue integrity. The immune system aims in the elimination of a foreign invader once activated the reaction cascades generate highly toxic products that damage any kind of cell. Based on the toxic potential and activity the discrimination between host cells (self) and microbes (foreign) is important. Thus the immune system serves four central functions (i) recognition of self or of foreign cells, (ii) discrimination between self and foreign, (iii) protection of self cells, and (iv) killing or elimination of foreign cells and microbes.

DNA Content Measurement for DNA Ploidy unit 75 and Cell Cycle Analysis

The second set of methods presented (see Basic Protocol 2 and Alternate Protocol 2) utilize detergents and or hypotonic solutions to permeabilize cells these methods generally provide more accurate estimates of DNA content compared to measurement of fixed cells. The approach presented in Basic Protocol 2 combines detergent treatment with use of proteolytic enzymes it is widely used for clinical material, especially for DNA analysis in samples of solid tumors. Alternate Protocol 2 is a simpler method designed for uniform populations (e.g., tissue culture cells).

Pyridinium Chlorochromate As a Urine Adulterant

Besides simple household chemicals, more sophisticated substances are advertised commercially as adulterants for urine drug tests. Wu et al. (13) reported that the active ingredient of Urine Luck is 200 mmol L of pyridinium chlorochromate (PCC). The authors reported that Urine Luck caused a decrease in response rate for all EMIT II drug screens and for the Abuscreen morphine and THC assays. In contrast, Abuscreen amphetamine assay produced a higher response rate, and no effect was observed on the results of BE and PCP. This adulteration of urine did not alter GC-MS confirmation of methamphetamine, BE, and PCP. However, apparent concentrations of opiates and THC were reduced.

Overview of Nucleic Acid Analysis

This chapter covers the methods most commonly used in analysis of nucleic acids. In unit 7.5, DNA analytical methods are presented that are adapted not only to DNA staining in fixed cells and in cells permeabilized by detergents, but also to supravital cell staining, staining of samples of cells isolated from archival material embedded in paraffin blocks, and detection of apoptotic cells characterized by fractional DNA content. A support protocol in unit 7.5 describes agarose gel electrophoresis of the degraded DNA, which is selectively extracted from the same apoptotic cells that can be identified by flow cytometry (Gong et al., 1994).

Pasteless Biopotential Electrodes

Hard anodization Super has been authorized as a coating for aluminum kitchen utensils, and it proves to be very stable even under high temperatures and the presence of corrosive substances used while cooking. The coating does not wear off with the use of abrasive scrubbing pads and detergents. These properties indicate that no toxic substances are released in the presence of heat, alkaline or acid solutions, and organic solvents. This makes its use safe as a material in direct contact with skin, and resistant to sweat, body oils, and erosion due to skin friction.

History of food irradiation

In 1958, when the Food, Drug, and Cosmetic Act was passed by the U.S. Congress, there were many unanswered questions. Would irradiated food be made radioactive What would be the effect of this additional radioactivity above that of the background on human health Would irradiation of food produce new toxic products such as carcinogens Would the process produce products with excessive Nutritional assessments showed that the irradiation process was no more destructive to nutrients than other processes then being used commercially. It was also demonstrated that there were no toxic products formed in quantities that would be hazardous to the health and well-being of consumers.

Sample Adulteration in Urine DOA Testing

The instant DOA testing procedures are instituted, opposing forces are at work to develop methods to avoid detection of drug use. Initially, common household chemicals such as laundry bleach, table salt, toilet-bowl cleaner, hand soap, and vinegar were used. More recently, a variety of products became commercially available, which can be ordered through Internet sites and tollfree telephone numbers. Commercially available adulteration products can be classified into two broad categories. The first category consists of specific fluids or tablets, which when taken along with plenty of water, serve to flush out drugs and metabolites, resulting in diluted urine and reduced concentrations of drugs or metabolites. Examples of products in this category include Absolute Detox XXL drink, Absolute Carbo Drinks, Ready Clean Drug Detox Drink, Fast Flush Capsules, and Ready Clean Gel Capsules. All products are available from Internet sites. Root Clean is a hair-cleansing system targeting drug...

Bile Secretion And Gall Bladder Function

The liver degrades many drugs and toxic products as well as a wide variety of hormones. Some are excreted in the bile. Others are made water soluble by conjugation to substances like glucuronic acid or by chemical transformation, and are then readily excreted by the kidneys.

Oxidative Stress Related Disorders

Oxidative stress is implicated in the inflammatory demy-elination that characterizes multiple sclerosis suggesting GST polymorphisms may be associated with disability. In 177 patients with disease duration over 10 years, GSTM3 AA (OR 2.4) and homozygosity for both GSTM1*0 and GSTP1*Ile105-encoding allele (OR 5.0) were linked with severe disability suggesting that long-term prognosis in MS is influenced by GST-mediated ability to remove toxic products of oxidative stress. 1 Exposure to ultraviolet radiation also results in local oxidative stress in skin. Response to such exposure, examined as minimal erythema dose, has been shown to be mediated by GSTM1 and GSTT1 genotype in a gene dosage-dependent manner. 19 Furthermore, nonmela-noma skin cancer has also been linked to these poly-

Method Specificity Selectivity

The FDA considers the removal of detergent residues an important aspect of the overall cleaning process 2 . If detergents are used to clean the manufacturing equipment, it is expected that they will be completely removed by rinsing, and that the removal be validated by testing for absence of residues. Unlike the situation with product residues it is not always as straightforward in selecting which component of the detergent to target for residue analysis. The pharmaceutical manufacturer may not have access to the specific composition of the detergent, although suppliers are usually willing to provide information on the chemical class of active agent(s). Moreover, there is commonly more than one component that is responsible for the cleaning action of the detergent product. It then becomes a question of whether to develop a test for a specific target component or for the whole detergent product. If choosing a single component, one should focus on testing for the particular component...

The Function Of Cxcr4 Receptor Depends On Lipid Raft Formation

Lipid rafts are membrane domains rich in sphingolipids and cholesterol, which form a lateral assembly in a saturated glycerophospholipid environment. The raft domains are known to serve as moving platforms on the cell surface, and are more ordered and resistant to nonionic detergents than other areas of a membrane40. These domains also act as good sites for crosstalk between various cellular proteins39,41. Lipid rafts have been shown to be important for T-cell

Effects on Particular Organs or Organ Systems

The eye is vulnerable to irritants such as smog, solvents, detergents, and corrosive substances. Other pollutants act systemically and can damage the optic nerve. For example, methanol and carbon disulfide damage the central vision in this way, and pentavalent arsenic and carbon monoxide affect peripheral vision.


Economically, the most significant single use of proteases is their incorporation into detergents 7 . They facilitate the removal of biological dirt, which is mainly protein-based. Obviously, the characteristics of the proteases must be compatible with standard washing conditions. Accordingly, the most successful detergent proteases are stable at alkaline pH values at relatively high temperatures and in the presence of bleach, surfactants, and sequestering agents. Screening of proteases from a wide range of sources has identified members of the bacterial subtilisin subfamily as the most appropriate for detergent application 15,16 . Typically, they are optimally active at temperatures between 45 to 65oC and in the pH range of 9 to 12. Many such subtilisins have been modified by genetic engineering with the general aims of increasing their thermal stability as well as improving their resistance to oxidation.

Enzyme Engineering

The past two decades have witnessed intensive industrial and academic research attempts to develop techniques to engineer enzymes to meet industrial demands, to replace traditional chemical catalysts used widely throughout industry, and to broaden the scope of enzyme applications 9,14,15 . Perhaps subtilisin, a bacterial serine protease, best illustrates the multiple goals of enzyme engineering. Subtilisin has been used for many years as a key component of many detergents, in addition to being used in many other applications in industry. Thus far, mutations in over 50 of the 275 amino acid (aa) residues of subtilisin have been reported in the literature, resulting in improvements in subtilisin's overall catalytic activity, mechanism of action, stability (thermal and pH), substrate specificity, surface activity, folding mechanisms, and in the evolution of new activities for subtilisin (see 24 and references therein for a comprehensive review).


Antimicrobials are chemicals that kill or inhibit the growth of microorganisms, depending on the use concentration and conditions. They include chemical preservatives and antiseptics, as well as drugs used in the treatment of infectious diseases of plants and animals. Two major types of antimicrobial agents are antiseptics and disinfectants. Antiseptics are designed to be applied on the skin, but should not be taken internally. Common examples are iodine solution, alcohols, detergents, and silver nitrate. Disinfectants are agents that kill microorganisms, but not necessarily their spores on inanimate surfaces. They are not generally safe for application to living tissues, but they may be applied on objects such as tables, floors, utensils, etc. Some examples are chlorine and its compounds, hypochlorites, copper sulfate, phenols, and quaternary ammonium compounds. In general, antimicrobial or anti-infective materials encompass a wide variety of substances including heavy metals...

The Virus

The family Astroviridae encompasses small nonenveloped viruses that infect a variety of animal species. They were originally classified among the small round structured viruses (SRSV) causing diarrhea in humans. They are round with icosahedral symmetry and 27-30 nm in diameter. They were first described by Madeley and Cosgrove 1 as a potential cause of diarrheal disease in human infants and named for the characteristic five- or six-pointed star (astron is Greek for a star), visible on the capsid surface by negative stain electron microscopy (Fig. 1). There are small surface projections consisting of 30 dimeric spikes protruding some 50 A from the virus surface. In 1981, serial passage of human astrovirus was achieved in primary human embryo kidney (HEK) cells by Lee and Kurtz, 2 but astrovirus is more conveniently cultured in CaCo-2 cells. However, trypsin must be included in the culture medium. 3 Astrovirus particles are stable at pH 3, but disassemble at pH 10.5 and are resistant to...


Compared with aqueous-based immunoassays, lateral-flow tests require that the capture reagents remain biologically active after being desiccated on the nitrocellulose for months to years. Many capture reagents meet this criterion, but not without consideration of the chemistry of the buffers used to apply them (10). In general, the reagent buffers should be kept as simple as possible (1) buffer molarity < 10 mM, (2) sodium chloride and other salts eliminated, (3) surfactants and detergents eliminated, and (4) other additives eliminated. Evaporation of the water used in the carrier buffer concentrates other solutes around and onto the capture reagent. The concentration of the buffer and other salts increases as a larger proportion of the water is evaporated, potentially denaturing proteins. The buffer and salts eventually crystallize and may mask the capture reagent. Larger crystals can also clog the pores of the membrane, retarding sample flow. In either case, the net effect is...


While a detailed treatment of cleaning supplies and methods is beyond the scope of this section, some discussion is warranted. Consult your microscope's manufacturer or local dealer for any booklets or written materials that might be available. Many service dealers can provide expert advice and tips, as well as proper cleaning supplies, so don't hesitate to consult them.

Endocrine disrupters

Returning to the problem of elevated levels of active hormones in the waterways, another aspect is that steroids do not occur in bacteria, although they are present in fungi, and so bacteria lack the necessary pathways to allow complete degradation of these hormones at a rate compatible with the dwell time in sewage treatment plants. The consequence has been raised levels of reactivated oestrogen and 17a-ethinyloestradiol in the waterways leading to disturbances of the endocrine, or hormonal, system in fauna downstream from sewage treatment plants. Such disturbances have been monitored by measuring the presence of the protein vitellogenin (Sole et al. 2001) which is a precursor to egg yolk protein, the results of which have indicated feminisation of male fish in many species including minnows, trout and flounders. The source of environmental oestrogens is not confined to outfall from sewage treatment plants, however, the fate of endocrine disrupters, examples of which are given in...


Colony PCR is a whole-cell-based assay, and thus will save a considerable amount of time, labor, and chemical reagents. The possibility of experimental errors or contamination is also reduced because DNA purification is not required. 3 A comparison study with two DNA extraction methods using mycobacterial isolates indicated that the heating method is more sensitive and no less specific than a conventional chemical method (phenolchloroform). The loss of DNA during purification and the presence of inhibitory substances are the most likely explanations for the lowered PCR sensitivity results obtained via the chemical method. 8,9 PCR is a very sensitive and rapid method of detecting specific DNA sequences in a wide variety of samples. The most common problem is contamination of reagents with target DNA. However, colony PCR will be performed on bacterial colonies directly obtained from broth culture or plates, which will lower the opportunity of contamination resulting from DNA...

Properties of Virion

The virions are 150-350 nm in diameter, pleomorphic, but usually spherical in shape. They consist of a nucleo-capsid surrounded by a lipid envelope. Virion Mr is around 500 x 106 Virion buoyant density in sucrose is 1.18-1.20gcm- . Some viruses (particularly of Pneumovirinae) are also produced in long filamentous form. Virions are highly sensitive to dehydration, heat, detergents, lipid

Spectrum of Activity

Acidified anionic detergents Materials compatibility constrains formulations to prevent corrosion, intended only for industrial sanitizing, not suitable for general hospital hygiene Volatile organic, flammability, potential for abuse, activity declines on dilution, does not clean and can fix material to surface, does not kill spores, may not be suitable for continuous exposure of some elastomers Potential carcinogens, skin and respiratory sensitizers, toxic residues, fixative especially problematic on prolonged exposure, some stain proteins, sporicidal only on prolonged exposure, noxious gas produced when mixed with chlorine Readily neutralized by organic materials, pungent irritating smell, toxic and mutagenic by-products, corrosivity and materials damage, bleaching inappropriately (chlorine), staining (iodine), allergic reactions (iodophors), efficacy and stability pH dependent Corrosivity and materials damage, irritant to skin and mucous membranes at high concentrations, can be...


Early research identified glucosylceramide as the compound that accumulated in Gaucher's cells but it was not until 1965 that the pioneering efforts of Brady, et al. 10,11 in the United States and Patrick 12 in the United Kingdom, revealed that a deficiency of the lysosomal enzyme P-glucocerebrosidase was the defect leading to the accumulation of glucosylceramide in Gaucher Disease. Following the identification of the defective enzyme, it was suggested, as early as 1966 13 that the disease may be treated by ERT. Several groups put a great deal of effort over the following years into purifying and characterizing the enzyme. Since we now know that P-glucocerebrosidase is hydrophobic in nature, unstable at neutral pH, and requires the presence of detergents for solubility and full activity 14-16 , it is not surprising that these early attempts to produce a pure preparation of the enzyme met with varying degrees of success 17-23 . In vitro studies have demonstrated that...


The metabolism of foreign compounds and many endogenous compounds depends on sulfate conjugation. Endogenous compounds include the biosynthesis of thyroid and steroid hormones, certain proteins, and peptides. Primary, secondary, and tertiary alcohols, phenols, and arylamines can form sulfate esters via sulfation. Sulfate esters are completely ionized and water soluble and quickly excreted by the organism. Although most compounds undergoing sulfation are converted to less toxic products, a few form reactive intermediates that have been implicated in carcinogenesis.


Although SPME has been widely used for highly efficient extraction of food components, little is known about the applicability of this technique for monitoring fragrance materials in household cleaning products, soaps, detergents, cosmetics, toiletries, etc. (61-63). Until recently, costly and laborious sample preparation techniques were required to isolate, separate, and identify fragrance components in perfumery matrices.

Molecular Biology

In vitro transcription catalyzed by permeabilized AMEV virions requires different conditions from those needed for transcription from VV virions. Unlike VV reactions, which use low levels of detergent and reducing agent, AMEV transcription requires higher levels of reducing agent and the requirement for detergent is less stringent, as transcription can occur in the absence of detergent but not in the absence of reducing agent. AMEV reactions are optimal when an ATP-generating system is present. Such a system is not required for VV virion-mediated transcription. This indicates that the structure of the AMEV virion is different in its response to both reducing agents and detergents. Our attempts to isolate AMEV cores with methodology optimized for the preparation of VV cores has also indicated that key AMEV core proteins partition differently from their VV counterparts.

Periapical Disease

Triggered by bacteria or their toxic products that diffuse through the apical foramen, the periapical tissues respond by chronic inflammation and bone resorption. In this way, a cavity is formed lying adjacent (Figs. 7.6a, b) to the root tip and filled with granulation tissue, the so-called apical granuloma. Also, cell rests of Malassez lying in or in the vicinity of the periapical area may show reactive proliferation leading to the formation of a cystic cavity, the radicular cyst. When the noxious stimulus is removed by cleaning and filling the pulpal space, the inflammation may subside and complete healing can occur. Sometimes however, the granulation tissue is not replaced by new periapical bone but by fibrosis in such instances a periapical scar is formed.


Aflatoxins are heat stable and easily transformed to toxic products. Treatment with ammonia reduces and inactivates aflatoxins. Lactic fermentation at pH < 4.0 results in the conversion of AFB1 to AFB2a, which is less toxic. Other environmental conditions, such as the presence of organic acid, also irreversibly convert AFB1 to aflatoxicol B, which is 18 times less toxic than AFB1. Detoxification results in the opening of the lactone ring (see Figure 13.2) and can be monitored by reduced fluorescence.

Plant Selection

It should be obvious that the major criteria for plant selection are the particular requirements for the method to be employed and the nature of the contaminants involved. For example, in the case of organic phytotransformation this means species of vegetation which are hardy and fast growing, easy to maintain, have a high transpiration pull and transform the pollutants present to nontoxic or less toxic products. In addition, for many such applications, deep rooting plants are particularly valuable.


Lister was the first to use carbolic acid in antiseptic methods (for instruments, ligatures, surgeon's hands, etc.) 29 . Today, ortho-phenylphenol and ortho-benzyl-para-chlorophenol are derivates of phenol that are used as constituents of hospital disinfectants 5,11 . Different studies have shown different results about the antimicrobial efficacy of these disinfectants. Studies by manufacturers demonstrated that commercial phenolics were bactericidal, fungicidal, virucidal, and tuberculocidal, but were not sporici-dal 5,11 . On the other side, some studies from independent laboratories demonstrated their antimicrobial ineffectiveness 30,31 . Anyway, they may be used for low- and intermediate-level of disinfection. They are commercially available with added detergents to provide one-step cleaning and disinfection.

Iodine and lodophors

Elemental iodine is normally used in an aqueous or alcoholic solution, but it often causes irritation and stains skin. Because of better acceptability, it has been replaced by iodophors, iodine complex with carriers, polyvinylpyrro-lidone (i.e., povidone) and ethoxylated nonionic detergents (i.e., poloxa-mers). The amount of molecular, free iodine present determines the level of antimicrobial activity of iodophors. Typical 10 povidone-iodine formulations contain 1 available iodine and yield free iodine concentrations of 1 ppm 61 . The concentration of iodine in antiseptic agents is much lower than in disinfectants.


Human PON1 is rather unstable, and tends to aggregate in the absence of detergents.52 Nor is it amenable to functional expression in bacteria or yeast, and thus to mutagenesis, library selection, and protein engineering. These factors led us to directly evolve PONs for bacterial expression and increased solubility.53 Family shuffling of four PON1 genes (human, rabbit, mouse and rat) (Fig. 6), followed by screening for esterolytic activity, led to recombinant PON1 variants (rePON1s) that are expressed well in E. coli. These variants diverged from the wild-type (wt) rabbit PON1 by 14-32 amino acids contributed by the other three PON1 genes. The rePON1 variants exhibited enzymatic properties essentially identical to those of the


One of the best and most widely used cationic antiseptics is chlorhexidine. It is used in the form of acetate, gluconate, or hydrochloride, either alone or in combination with other antiseptic agents such as cetrimide, alcohol, etc. Chlorhexidine gluconate was introduced as an antiseptic in the United States during 1970s, several decades after approval for use in Europe and Canada 51 . Preparations of chlorhexidine are available as aqueous and alcoholic solutions, detergents, or powders 49 .


Triclosan is a chlorinated bisphenol, which dissolves well in some detergents such as anionic soaps, and alcohol. It is only sparingly soluble in water. Concentration of 0.2 -2 are effective against gram-positive bacteria and gram-negative bacteria but with variable and poor activity against Pseudomonas aeruginosa. This agent is also active against mycobacteria and Candida spp., but it has limited activity against filamentous fungi such as Aspergillus spp. 49,72 . Because of acceptable antimicrobial activity, use of triclosan has risen dramatically in the past few years. Today it is used as an additive to hand soaps, dish-washing products, cosmetics, and toothpaste. It is also used as an additive to plastics, polymers, and textile and implant devices to confer these materials with antibacterial properties.


Surface active agents or surfactants, such as soaps and detergents, have the ability to orientate themselves between two interfaces to bring them into closer contact (Figure 13.6). The value of soap has less to do with its disinfectant properties than with ability to facilitate the mechanical removal of dirt and microorganisms. It does this by emulsifying oil secretions, allowing the debris to be rinsed away. Detergents may be anionic (negatively charged), cationic (positively charged) or non-ionic. Cationic detergents such as quaternary ammonium compounds (ammonium chloride with each hydrogen replaced by an organic group, Figure 13.7) act by combining with phospholipids to disrupt cell membranes and affect cellular permeability.

Skin Tests

The semi-open test as described by Dooms-Goossens 6 is particularly helpful if strong irritancy under occlusion is suspected, e.g., in the case of shampoos, liquid soaps, nail varnish, and also industrial products such as glues, paints, inks, varnishes, etc. The golden rule is that when a subject comes into direct skin contact with such a product (either on purpose, e.g., cleaning products, or accidentally, e.g., soluble oils, paints), then the product may be tested in this way. Corrosive or other toxic materials (pH < 3 or > 10) that are normally used in closed systems only or with protection from appropriate clothing are excluded from testing. The material is applied to the skin with a cotton swab (about 15 l) on a small area (2 x 2 cm), left to dry (possibly dabbing with another Q-tip or tissue), and is then covered with acrylic tape (e.g., Micropore, 3M) (Fig. 2).

False Negatives

False negatives resulting from reaction inhibition have been widely reported and reviewed (30-32). Inhibition may be total or partial and can become apparent as complete reaction failure or as reduced sensitivity of detection. In some cases, inhibition may be the cause of false-negative reactions because workers have not incorporated internal controls in each reaction tube. Early evidence of exquisite sensitivity using mammalian cells (33) involving detection of a single molecule of DNA from a hair was not followed by similar sensitivity when PCR was applied to many microbial (and some mammalian) situations where poor sensitivity, specificity, and reproducibility have been reported (21,34-38). As discussed in detail in the following sections, there may also be potentially important effects in PCR typing reactions (39), and difficulties can occur in post-PCR manipulation (40). Although systematic study of inhibition has seldom been the focus of published investigations, many workers...


Thus, hyperthermophilic extremozymes have potential applications in many industries, offering amylases for confectionary or alcohol production, proteases for amino acid production, baking, brewing and detergents, xylanases for paper Other extremophiles could also have roles to play. Psychrophiles may yield enzymes which will function at the low refrigerator temperatures typically required to avoid spoilage in food processing, for enhanced cold-wash 'biological' washing powders and in perfume manufacture, reducing evaporative fragrance losses. A use has been suggested for halophile enzymes in increasing the amount of crude oil extracted from wells, though whether this will ever be a commercial reality remains to be seen and, moreover, leaves aside any consideration of the 'environmental' aspects of increased fossil fuel extraction. Acidophilic extremozymes may one day form catalysts in chemical syntheses in acid solution, and alkaliphile derived proteases and lipases may replace...

Chemical bonds

Parts are excluded by the water and group together as described above, leaving the polar groups pointing outwards into the water, where they are attracted by hydrophilic forces. Detergents exert their action by trapping insoluble grease inside the centre of a micelle, while interaction with water allows them to be rinsed away.

DNA Purification

DNA can be bound to glass, silica particles, or other polymer surfaces in the presence of alcohol, high salt, or chaotropic agents and subsequently is released in low-salt buffers (26). Other approaches using detergents (27) or polyethylene glycol (28) to bind DNA onto polymer surfaces also have been developed. The solid-phase principle has been applied in several formats, such as cartridges, filters, and paramagnetic beads. Paramagnetic beads have the advantage over other solid phases that they can easily be manipulated by a magnet and thus eliminate the need for centrifugation steps and speeding up washing steps.

Amitava Dasgupta

Persons abusing drugs attempt to adulterate urine specimens in order to beat drug testing. Dilution of urine in vivo by consuming excess fluid and various detoxifying agents available through the Internet is a common practice. Household chemicals such as bleach, acid, table salt, laundry detergent, toilet-bowl cleaner, vinegar, lemon juice, and Visine (Pfizer) eye drops are also used for adulterating urine specimens. Most of these adulterants except Visine eye drops can be detected by routine specimen integrity tests (creatinine, pH, temperature, and specific gravity). However, certain adulterants, such as Klear , Whizzies, Urine Luck , and Stealth , cannot be detected by using routine specimen integrity testing. These adulterants can successfully mask drug testing if the concentrations of certain abused drugs are moderate. Several spot tests have been described in the literature to detect the presence of such adulterants in urine. More recently, urine dipsticks are commercially...

Healthy Chemistry For Optimal Health

Healthy Chemistry For Optimal Health

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