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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. Read more...

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Manufacturing Cleaning And Chemical Products

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Isolation of Detergents and H202 Resistant Alkaline Proteases

Alkaline proteases are used extensively in detergents, the food industry, and leather tanning. Enzymes produced commercially are derived only from microorganisms, and the microorganisms must be able to produce a high enzyme yield from low-cost substrates. The success of alkaline proteases in detergents is depend on whether the enzymes have the following properties 1) a wide pH activity range, 2) stability under high alkaline conditions, 3) high activity and stability in the presence of surfactants, 4) high stability in the presence of builders such as chelating reagents and bleaching agents, 5) high activity over a wide temperature range, 6) long shelf-life, and 7) low production cost. Although many enzymes have been reported, the alkaline proteases described above, however, have several weak points in their enzymatic properties, e.g., they are sensitive in the presence of oxidants and chelating agents. These disadvantages have been overcome by the isolation of new Bacillus strains by...

ATPS for Product Recovery in Biotechnology

Isolation of membrane proteins, normally a rather difficult and time-consuming task, has been successfully achieved by the convenient two-phase extraction procedure. Use of nonionic detergents, e.g., Triton X-114, which separate at rather mild temperatures like 20 C into a detergent-rich and detergent-poor phase, provide novel means for obtaining enriched preparations of integral membrane proteins from complex biological systems (20,21). 4. Two-phase partitioning also holds promise for rapid isolation of DNA. By using novel phase systems made of PEG-salt containing chaotropic agents and detergents, it has been shown that nucleic acids partition with high yields to the salt-rich phase, whereas proteins and other cellular material are concentrated in the other phase or precipitates at the interface (22).

Biochemistry of asynuclein aggregation

A-Synuclein is a small 140 amino acid, natively unfolded protein with little secondary or tertiary structure whose aberrant aggregation has been linked to the etiology of PD. a-Synuclein can assume either an a-helical conformation upon binding to lipids and membranes or a b-sheet conformation upon aggregation at high concentrations, elevated temperature, agitation, or in the presence of metals, simple alcohols, or detergents 67,68 . Increases in b-sheet content are associated with formation of small soluble oligomers, larger protofibrils, and the macro-molecular fibrils. Dimers are the smallest oligomeric aggregates with limited b-sheet structure and have been proposed to act as seeds in the nucleation-dependent aggregation of a-synuclein 69,70 . Both oligomers and protofibrils have increased amounts of b-sheet structure compared with monomeric or lipid bound a-synuclein. The predominantly observed structure of the protofibrils is globular, although they are also able to form rod-like...

Principles of Selected Molecular Biology Techniques

Classical techniques are based on lysing cells with lysozyme, alkali, or detergents. The removal of protein and other contaminants is effected by incubation with protease or by extraction with phenol or chloroform. The extract is concentrated by precipitation with ethanol in presence of sodium or ammonium acetate. If necessary, RNA can be removed, using DNase-free RNase. The advantage of using proteinase K is that, in addition to releasing DNA from chromatin, it also destroys nucleases, which otherwise would reduce the average molecular weight of DNA (N3). However, after the lysis of cells has been accomplished, proteinase K has to be removed before the isolated DNA can be subjected to restriction enzyme cleavage. Removal of proteinase K is effected by phenol chloroform extractions, which leave DNA in the aqueous phase while trapping proteins and lipids in the organic phase. Proteinase K removal can be performed on DNA isolated on a 96-well tissue culture plate (Rl). All steps from...

B2 Commercial Protein Estimation Kits

One of the most used kits is the DC protein estimation kits from Bio-Rad. The assay is sensitive and is also reliable in the presence of reducing agents and detergents. The assay is a modified Lowry method. This method is rapid and the reading can be done in 15 min with the color change not varying with time, making the reading stable and reliable. The reaction involves protein with the alkaline copper tartrate and the Folin reagent. The protein residues, especially, tyrosine and tryptophan, and to a lesser extent, cysteine cysteine and histidine, reduces the Folins reagent producing a blue color.

Bacterial Cell Lysis and Release of Intact Chromosomal DNA

A pure culture of a bacterial isolate of known identity is incubated overnight in a nutrient broth, such as trypticase soy broth (BD Biosciences, Sparks, MD, USA) in order to achieve 109cells mL, which is the concentration needed to obtain a visible band pattern. Standard DNA extraction procedures are inappropriate for the analysis of large chromosomal DNA molecules because of DNA shearing caused by the application of mechanical force in the protocol. In order to prevent DNA damage, it is important to mix intact bacterial cells with warmed, liquid-phase agarose, and this mixture can then be pipetted into plastic molds to form 10 x 5 x 1.5 mm agarose plugs. The whole cells embedded in plugs are lysed and deproteinized by detergents and enzymes (e.g., lysostaphin, lysozyme, proteinase K, mutanolysin or lyticase) in situ (Table 9.1). Following cell lysis, the plugs are washed 4-5 times with wash buffer containing 20 mM Tris and 50 mM EDTA (pH 8.0) to remove cell debris and proteinase....


Blepharitis is inflammation of the eyelid margins that is most often misdiagnosed as an ocular allergy because it commonly causes conjunctivitis as well. Infection or seborrhea are common causes. As in patients with atopic dermatitis, the most important organism isolated from the lid margin is Staphylococcus aureus. Antigenic products and not the colonization itself are thought to play the primary role in the induction of chronic eczema of the eyelid margins. The symptoms include persistent burning, itching, tearing, and a feeling of dryness. Patients commonly complain of more symptoms in the morning than in the evening. This is in contradistinction to patients with dry eye syndromes, who complain of more symptoms in the evening than in the morning because of drying out of the tear film during the day. The crusted exudate that develops in these patients may cause the eye to be glued shut when the patient awakens in the morning. The signs of staphylococcal blepharitis include dilated...

In Search of the Mechanism of Persister Formation

Identification of the mechanism of persister formation presents a formidable challenge due to an apparent redundancy of persister genes. Thus, attempts to identify persister genes by screening transposon insertion libraries for either increased or decreased survival to antibiotics were not successful (Hu and Coates 2005 Spoering et al. 2006). In a recent report, phoU was identified as a putative persister gene using a similar approach. However, the phoU mutant had a decreased MIC to a number of antibiotics. This suggests that phoU Tn is a pleiotropic mutation. It is interesting to compare this experience with identifying genes controlling another function that produces dormant cells sporulation. It is easy to obtain specific spo mutants specifically lacking the ability to make spores from a knockout library (*), and this is indeed how most spo genes were identified. Genes controlling tolerance resemble in this regard those coding for multidrug resistance pumps (MDRs). In P....

Mechanisms Of Resistance To Specific Drugs

To multiple antimicrobial drug classes is multifactorial. Most small hydrophilic molecules, including antibiotics, enter the cell via nonspecific porin channels present in the outer membrane. The outer membrane permeability of P. aeruginosa has been shown to be 10- to 100-fold less efficient than, e.g., in Escherichia coli (28). This high intrinsic antibiotic resistance that typifies P. aeruginosa is recognized to be the result of synergy between this highly impermeable outer membrane and the expression of broad-spectrum multidrug efflux systems (Table 2). These efflux systems serve the physiological function of preventing toxic compounds from entering the cell in significant concentrations. They actively remove biocides, dyes, detergents, organic solvents, and metabolites as well as antibiotics from the cytoplasm or periplasmic space (29). The most important and well-studied family of these is the resistance-nodulation-cell division (RND)-type family of pumps. There are, in addition,...

Hans Erik Akerlund 1 Introduction

To understand the function of a biological membrane like that of chloroplast thylakoids, it is important to understand the arrangement of its different protein and lipid components. Preparations that have proven to be particularly suited for such studies are those consisting of membrane vesicles that are turned inside-out. Inside-out vesicles from the thylakoid membrane were first obtained from spinach chloroplasts by a combination of mechanical fragmentation and separation by aqueous two-phase partition (1,2). By the same or very similar procedures, inside-out thylakoid vesicles have now also been obtained from other plant sources such as pea (3), barley (4), mangrove (Avicennia marina) (5), lettuce (6), Euglena gracilis (7), cyanobacteria (8,9) and the photosynthetic bacteria Rhodopseudomonas viridis (10). Because the isolation procedure does not involve the use of detergents, the inside-out thyla-koids have a preserved membrane structure and are ideally suited for...

Role of Environmental Factors

Certain laundry detergents, bleaches, soaps and household cleaning chemicals act as irritants for patients with AD. Mild laundry detergents without bleach are generally better tolerated. Washing clothing through a rinse cycle twice usually ensures removal of the detergent and may be beneficial in some sensitive patients. Mild skin soaps should also be used for bathing by AD patients. They are generally less drying, less irritating and less likely to induce pruritus. Skin should be protected from household cleaners by wearing protective gloves or clothing. The skin barrier is frequently altered in AD and will not withstand the general intrusions that normal skin can endure.

Two Dimension Sdspage Analysis

When preparing protein extracts for isoelectric focusing, it is best to avoid solutions with high ionic strength and ionic detergents such as SDS. High salt and detergent content interfere with the initial phase of 2D electrophoresis and proteins do not separate or focus properly. Also, during sample preparation, the removal of nucleic acids and or cellular debris improves protein separation and decrease background interference for visualization. Protocol after Kashem et al. (2007) for 2D SDS-PAGE

Environmental Control

Clothing should be loose and free of wool. Cotton fabrics are generally the best tolerated. Coarse fabrics in clothing and bedding should be avoided. Complete rinsing of detergents, soaps and bleach from clothing and bedding will also minimize their irritant potential. Occupational aggravating agents such as chemicals, irritants and solvents should be avoided by older patients with AD. Minimization of emotional stress will also lessen the potential for pruritus.

Isolation of Alkaline Proteases for Laundry Detergent Additives

Kobayashi et al. (1995) isolated M-protease that was suitable for use in detergents from alkaliphilic Bacillus sp. KSM-K16. The M-protease was purified to homogeneity from the culture broth by column chromatographies. The N-terminal amino acid sequence was The molecular mass of the protease was 28 kDa, and its isoelectric point was close to pH 10.6. Maximum activity toward casein was observed at 55 C, as shown in Fig. 7.4. The activity was inhibited by phenylmethylsulfonyl fluoride and chymo-statin. The enzyme was very stable in long-term incubation with liquid detergents at 40 C. This protease exhibited good properties as a laundry detergent additive except for oxidant resistance. The gene responsible for the M protein was cloned and sequenced. The accession number is Q99405.

Environmental Cleaning Interventions

This means cleaning horizontal (high-touch) surfaces that commonly harbor C. difficile spores (e.g., bedrails, call buttons, telephones, floors) more often than upon terminal cleaning of the room. In Canada, a best practices document actually recommends twice daily cleanings in healthcare facilities. Due to staffing inadequacies faced by hospitals today, environmental services departments may have altered important cleaning practices unbeknownst to the infectious diseases experts within the facility. Failure to clean high-touch surfaces with the appropriate agent may lead to increased transmission within an institution. When faced with an outbreak (or even as a preventative measure), multidisciplinary discussions that include environmental services leadership should take place to determine what cleaning products are being used, what surfaces are being cleaned, and how often. Bundled approaches where resources are dedicated toward environmental...

El Enzyme of Bacillus cohnii D6

In 1972, Yoshida and Horikoshi discovered a very- stable alkaline protease in the presence of detergents containing high concentration of perborate in the absence of calcium ion, (Japanese patent JP 740710). Bacillus sp. No. D-6 (FERM No. 1592, later designated Bacillus cohnii D-6, FERM P-1592) produced an alkaline protease, E-l, that was more stable in the presence of detergent additives at 60 C than Bacillus clausii 221 protease. Several properties are presented in Table 7.1. If the enzyme productivity could be increased, the E-l enzyme would be the best enzyme for detergent additives. After our discovery, many researchers tried to industrialize it, but without success. Almost thirty years later, Saeki and our colleagues (2000) dramatically increased the productivity to more than 10 grams liter using gene technology.

Antiintracellular Adhesion Molecule1 Enlimomab

Returning blood flow carries white blood cells (leukocytes) to small vessels. In this process, the leukocytes can occlude the vessels and cause additional ischemia. The leukocytes also release toxic products that directly injure endothelial cells and cause the formation of free radicals and cytokines. An agent, such as a monoclonal antibody (mab) to intracellular adhesion molecule-1 (ICAM-1), which inhibits leukocyte adhesion to vessel walls, prevents leukocyte adhesion and infiltration and, in animal models, limits the extent of reperfusion injury (4).

Size fractionation of viral structural proteins

Once pure virus is obtained, gentle disruption of the virions with mild detergents or appropriate salt treatments can lead to disruption of the particle and solubilization of the components. Proteins and nucleic acids can be separated from each other by a variety of extraction or differential degradation regimens. For example, small amounts of nuclease could be used to digest nucleic acid into nucleotides, or proteases could be used to digest proteins. These macromo-lecular components then can be separated according to size or charge, or a combination of both.

DNA Preparation and Hybridization

DNA quality has a great influence on the outcome of an array CGH experiment. In general, oligonucleotide platforms, especially when used for one-color experiments, are more sensitive to compromised DNA quality than are BAC arrays. One source of trouble can come from the DNA isolation procedure, for example, detergents that could hamper the subsequent labeling process and introduce noise in the data. Although such problems can be avoided quite easily by improving or changing the isolation protocol, the situation is much more difficult when it comes to retrospective studies using formalin-fixed, paraffin-embedded (FFPE) material. This kind of fixation very frequently results in damage of DNA.19 To cope with such damage, several adaptations of the DNA isolation and labeling protocols are required. Modifications can include the prolongation of tissue digestion time, with adding fresh proteinase each day, or the switch from enzymatic to chemical labeling systems. For other applications,...

Reverse Transcription

Reverse transcription normally begins soon after entry of the virion core into the cytoplasm of the infected cell. The reaction takes place in a large complex, roughly resembling the virion core, and containing Gag proteins including NC, RT, IN, and the viral RNA (23). The signal that triggers the onset of DNA synthesis is not known, though it may be as simple as the exposure of the viral core to the relatively high levels of deoxyribonucleotides present in the cytoplasm. This notion is consistent with the observation that simply stripping or permeabilizing the virion membrane with detergents in the presence of deoxyribonucleotides is sufScient to induce DNA synthesis. This may also be at least part of the explanation for the difSculty HIV has in completing reverse transcription and infection in quiescent cells (24). In some cells, notably cells arrested by starvation, triphosphate levels may be low and limiting for reverse transcriptase, so that addition of exogenous nucleosides can...

Pullulandegrading Enzymes

The demand for liquefying a-amvlase for use in laundry and automatic dishwashing detergents has been growing for several years. Detergent enzymes must be able to function in washing machines or dishwashers under conditions that are very unfavorable for the stability of the enzyme. The pH is high alkaline in washing conditions. The high temperature (55-60 C) in a dishwasher requires thermostable enzymes. Enzymes are preferentially resistant to various detergent ingredients, such as sufactants, chelators and oxidants. However, most bacterial liquefying a-amylases, such as Bacillus amy-loliquefaciens (BAA) and Geobacillus stearothermophilus (BSA), including Bacillus licheniformis (BLA), have optimal pH of between 5 and 7.5. Therefore, they are not practically suitable for use in laundry and dishwashing detergents with high alkalinity.

Pullulanase of Alkaliphiles Isolated by Kao Corporations Group

In screening alkaline cellulases for detergent additives, Ara et al. (1992) and Igarashi et al. (1992) isolated a novel alkaline pullulanase from alkaliphilic Bacillus sp. KSM-1876, which was identified as a relative of Bacillus circulans. The enzyme had an optimum pH for enzyme action of around 10.0-10.5, the highest pH for optimum pullulanase activity. This enzyme is a good candidate for use as detergent additives in dish washers, especially to remove starch from dishes in the presence of detergents. The gene (Accession Number AB049812) was isolated from KSM-1876. However, the plasmid-borne enzyme was not thermostable and not alkaline pullulanase, having an optimum pH of 7.5. The enzyme hydrolyzed pullulan 3.0-fold faster than soluble starch, but hydrolyzed amylose and amylopectin less efficiently. Therefore, the gene cloned was not the major alkaline pullulanase gene, but a minor one.

Fluorescence In Situ Hybridization FISH

The goal of this step is to prepare the cells in the specimen so that the probe can efficiently hybridize to its cellular DNA target but do so without significantly disrupting the morphology of the cells. Detailed protocols for prehybridization and hybridization can be found elsewhere.10,11 The pretreatment required for different specimen types (e.g., fresh cells vs. paraffin-embedded tissue) is similar but frequently has some important differences. For example, certain specimens such as paraffin-embedded tissue generally need to be treated with a protease such as pepsin to increase the accessibility of the probe DNA to its nuclear DNA target. It is important to not over- or underdigest the tissue with pepsin. Overdigestion can lead to a decrease in signal intensity and destroy nuclear morphology. Underdigestion can lead to auto-fluorescence, and an underestimation of signal copy number may result. Other methods that have been used to facilitate entry of the probe into the cell...

An Additive or Synergistic Interaction

With regard to so-called synergy, the only clinically well-proven example is for enterococcal endocarditis (an anti-enterococcal cell wall-active drug plus an aminoglycoside) (86). Additionally, synergy, although not well-proven clinically, has been employed in patients with pulmonary exacerbations of cystic fibrosis (CF) for Gram-negative-directed therapy (87). In this example, synergy studies are sent out and performed on isolates collected from the CF patient to determine the best combination of drugs to use. This is a relatively unique therapeutic situation, as the organisms are typically pan-resistant and the premise for using synergistic combinations is to lower the overall minimum inhibitory concentration (MIC). Since the goal of this treatment is to reduce the burden of the infecting organisms because eradication of the organisms is typically not possible and the host is different, this type of synergy cannot be extrapolated to non-CF patients who develop pneumonia. For MDR...

Keith Poole and Ramakrishnan Srikumar 1 Introduction

The broad substrate specificity of the multidrug resistance (MDR) efflux systems, which also accommodate a range of dyes, detergents, and organic solvents (9,10), in addition to antibiotics, means that it is also possible to measure the efflux activity of such systems directly using fluorescent dyes as indicator substrates. Coupled with a fluorescence spectrophotometer, this permits the continuous monitoring of drug efflux, and doesn't require the sampling of cells at different time points to assess drug retention expulsion over time, the latter being necessary when using radiolabeled antibiotics. The use of dyes to asses efflux is, obviously, limited to MDR systems which accommodate them, and cannot be used to monitor export by agent-specific efflux systems. Still, the ease of the assay, which avoids the need to separate cells from the radiolabeled drug in the growth medium (by filtration or cen-trifugation through silicone oil) and the attendant problems of background and low...

Discussion Of Assay Limitations

Other methods have been employed to monitor Bcl-2 family protein activities or interactions. Physical measurements can be used to monitor the effects of the Bcl-2 family members on lipid micelles. Bcl-2 (41,42), Bcl-xL (43), Bax (42,44-46), and Bid (47) form pores in lipid membranes micelles that are quantifiable by electrophysical methods or by monitoring release of small molecules (e.g., fluorescein). In many instances the formation of pores is greatest under rather acidic conditions, making the biological significance of these measurements uncertain. Monitoring dimerization of members of the Bcl-2 family has been proposed as a screening tool for examining small molecules that interfere with dimerization. Dimerization between Bcl-2 family members has been examined by surface plasmon resonance (48) and in ELISA formats that detect both partners of the dimer (46,49). However, great care must be exercised in interpreting the results of dimerization assays. Detergents have been found to...

Short Format for Cytochrome C Release Measurements

The cytochrome c ELISA kit 96-well plate was incubated three times with PBS for 10 min per incubation. To each well was added 32 L of buffer F containing 20 g of BSA. HRP-conjugated detection antibody in conjugate diluent free of detergents was added to obtain the desired concentration. Then Bcl-2 family proteins diluted in buffer E were added. The volume of each well was adjusted to 42 L with buffer E. Mitochondria were diluted into buffer C, and 8 L (approx 1.8 g of protein) was added to each well. The plate was floated in a 30oC water bath for the desired time. Then 200 L of buffer E were added and the wells were washed 5 times with 300 L per wash of buffer E and 100 L of color development reagents was added. After 3 min, stop solution was added and absorbance was measured at 450 nm using 540 nm as a reference wavelength. To determine total detectable cytochrome c, Triton X-100 was added to mitochondria to a final concentration of 0.1 .

Other Animal Exposures

Given our present understanding, it would be beneficial for patients with mouse and rat sensitization to minimize their exposure. Our knowledge regarding methods to decrease these allergens is currently lacking, but they are now being studied. A recent study demonstrated that certain measures are effective in lowering Mus m 1 in mouse-infested, inner-city homes. These interventions included filling holes and cracks with copper mesh and caulk sealant, vacuuming with HEPA filters, and cleaning surfaces with mild detergents. Traps and low-toxicity pesticides from professional exterminators were also used. Studies with large numbers of patients will be required to help determine the duration and degree of the clinical effect of these methods.

Morphological Changes Upon Virion Maturation

Mutant viruses lacking the protease show little change in morphology. Thus, cleavage of Gag and Gag-Pol is required for the restructuring of the virion into the mature form (395). The changes in morphology visible in electron micrographs are probably associated with major rearrangements of the Gag proteins. Indeed, the physical properties of the virus change dramatically upon maturation. Whereas the immature core is quite stable to non-ionic detergents and harsh conditions, the mature virion core is relatively labile. This change may reBect the inability of the immature virion, and the acquired ability of the mature virion, to uncoat upon infection of new cells and initiate reverse transcription.

The Properties Of Lipid Rafts

In terms of their role in immune cell signaling, the central feature of lipid rafts is that they provide a mechanism for the lateral segregation of proteins within the plasma membrane (3). This ability to segregate proteins provides a means of concentrating and compartmentalizing certain components of signaling pathway and excluding others. At present, the rules that govern the inclusion and exclusion of proteins from lipid rafts have not been fully delineated, however, analyses of the composition of rafts allows some generalizations to be made. For their characterization, lipid rafts have been separated from the glycerolphospholipid membranes in cells based on their differential solubility in nonionic detergents (3,7). Thus, for example, lipid rafts are isolated from B lymphocytes by sucrose density gradient sedimentation of cell lysates prepared in 1 Triton x 100 at 4 C, conditions under which lipid rafts are insoluble. There are a number of potential pitfalls associated with the...

Nonspecific methods of introducing viral genomes into cells

A similarly inefficient and nonspecific process called transfection is often used to introduce viral genomes (especially DNA genomes) into cells. Isolated genomes can be aggregated into the proper-sized particles by precipitation into aggregates using calcium phosphate (Ca3(PO4)2), and cells can be treated to readily incorporate the aggregates. Alternatively, viral genomes can be concentrated inside lipid vesicles called liposomes in solution and these can be readily assimilated by cells that have been specifically treated with mild detergents so that their plasma membrane can fuse with the liposome. Several other methods of introducing DNA, RNA, or even proteins into cells have also been effectively exploited. Transfection of plants can be efficiently carried out by literally shooting microscopic pellets of plastic coated with the appropriate macromolecular concoction using an air blast. Macromolecules can also be introduced into cells using electric fields. In all these cases the...

Torsten Minuth 1 Introduction

Cloud Point Extraction

Extraction systems based on nonionic surfactants have been described as an alternative to the standard polymer polymer or polymer salt systems. Phase-forming surfactants are, for example, the nonionic polyoxyethylene-type detergents. This kind of aqueous two-phase system (ATPS) is simply induced by a switch in the temperature on the basis of the temperature-dependent reversible hydration dehydration of the polar ethylene oxide headgroups. A single isotropic micellar phase separates into two isotropic phases one of the resulting two aqueous phases, the so-called coazervate phase, is enriched in detergent, whereas the other is depleted (1). The detergent forms micelles in the detergent-depleted phase and is believed to exist in the form of lamellar stacks in the coazervate phase (2). Both phases have a high water content. The temperature at which the phase separation occurs is referred to as the cloud-point. The clouding temperature depends on the structure of the polyoxyethylated...

Cleaning or disinfection of environment

Routine cleaning of environmental surfaces with detergents and elimination of heavy dust is sufficient in most circumstances (WIP, 2002 Module 6.4). Detergents, disinfectants, and cleaning equipment itself may become a source of contamination. Werry et al. (1988) reported contamination of detergent solutions used for cleaning of surfaces. The contaminants, mainly Gram-negative non-fermentative bacilli, including Acinetobacter spp. and Pseudomonas aeruginosa. Since contamination may occur during the preparation of fresh solutions, cleaning solutions must be prepared daily or as needed, and frequently be replaced with fresh solution according to facility policies. The mop head should be changed at the beginning of each day and or after cleaning up large spills of blood or other body substances. The use of disposable materials is preferred for all methods of cleaning. When using non-disposable materials, they have to be sent to the laundry service immediately after completing a cleaning...

Active Immunization

Vaccines are preparations, administered either orally or parenterally, which stimulate a protective specific immune response in the recipient without themselves causing disease. With some vaccines (e.g. hepatitis B) it is necessary to add an adjuvant which non-specifically potentiates the immune response. Adjuvants, usually aluminium salts, can also delay the release of vaccine material from the injection site. In principle there are two main types of viral vaccines available, namely the attenuated live virus vaccine and the non-replicating ('killed') variety. In the first instance virus is manipulated in vitro to be of low virulence but able to replicate in the vaccinee and stimulate the desired protective immune response without causing disease. In the case of non-replicating vaccines the virus may be replicated in an appropriate laboratory system, e.g. cell culture or embryonated eggs, purified and finally inactivated ('killed') by chemical means (usually formalin). A further...


The importance of GSH for neutralization of toxic products of lipid peroxidation is confirmed by the experiments with SDI, which effectively (by 90.8 ) inhibited increased flux through sorbitol dehydrogenase in the 3-week streptozotocin-diabetic rat model. GSH depletion in 3-week streptozo-tocin-diabetic rats was further exacerbated by the SDI treatment, which is consistent with a further accumulation of HNE over the level in the untreated diabetic group. A more advanced GSH depletion in the SDI-treated diabetic rats compared with the corresponding untreated group is consistent with the findings of Geisen et al. (32) for diabetic lens. These findings implicate sorbitol accumulation-linked osmotic stress in the mechanisms underlying increased vulnerability of diabetic peripheral nerve to oxidative injury and are in accordance with other reports demonstrating the inconsistency between a substantial ( 40 ) depletion of GSH versus a very minor (2) or an absent (12) depletion of GSSG,...

Skin Diseases

Contact dermatitis, both irritant and allergic, can be seen in infants and young children. The skin eruption of irritant dermatitis varies with etiological agent, but is commonly seen on the cheeks, chin, extensor surfaces of the extremities, and the diaper area. Irritant dermatitis is typically less pruritic than AD and improves with removal of the irritant (i.e., soaps, detergents, abrasive bedding). Allergic contact dermatitis is characterized by a pruritic, erythematous papulovescicular eruption that involves exposed areas of contact. This dermatitis is uncommon during the first few months of life and can frequently be delineated by a careful history.


Irritant reactions tend to occur often within minutes, burn rather than itch, and heal rapidly on avoidance. Irritant patch test responses typically are evident within a few minutes, although they may be cumulative or even delayed. They are often somewhat dose-related, disappearing on dilution. They are often sharply marginated, and they occur on first exposure so that prior sensitization is unnecessary. Irritant reactions from soap, detergents, and solvents are often shiny, dry, and fissured.


Cleaning is the removal of all visible and invisible organic material (e.g., soil) from objects to prevent microorganisms from thriving, multiplying, and spreading. It is accomplished using water with detergents or enzymatic products. In sanitary facilities (e.g., washbasins, toilets), separate buckets and cloths have to be used and alkaline detergent is recommended for cleaning.

Hair shafts

Because the hair shaft contains very low levels of DNA it is prone to contamination but unlike many other types of biological evidence with low levels of DNA it is possible to clean the hair shaft prior to DNA extraction. Several methods have been used to clean hair including washing in mild detergents, water and ethanol and also using a mild lysis step in the same way as is used in the differential extraction of semen 21 .

Lower Extremities

On the legs and thighs, contact dermatitis may be from nickel or phosphorus sesquisulfide in matches, rubber or dyes in stockings, detergents, fabric finishes, and clothing (Fig. 13) even reactions from epoxy in knee pads have been reported. The differential diagnosis includes nummular, atopic, and stasis eczema (Fig. 14), poison ivy dermatitis (Figs. 3-5), contact from medications (Figs. 8 and 9), other eczemas, and many other dermatological diseases.

Penile Disorders

Several types of inflammation problems may involve the penis and the urethra. Balanitis occurs when the glans, or head, of the penis becomes red and sore. Usually the cause is unknown, but it is sometimes caused by urinary tract infection or allergic reactions to clothing or detergents. In uncircumcised men, the irritation may result when the foreskin is narrow or difficult to retract, and secretions become trapped beneath the foreskin.

The Technology

The probes for cDNA arrays are normally polymerase chain reaction (PCR) products of 0.6-2.4 kb in length 25 , generated using vector- or gene-specific primers from cDNA libraries or clone collections. Once prepared and appropriately cleaned up, the cDNA probes are printed onto the substrate at defined locations. cDNA probes generated by PCR need partial purification to remove unwanted contaminants such as salts, detergents, PCR primers, and proteins which are present in the PCR reaction mixture. The cDNA spots typically contain a few nanoliters of a product at 100- to 500-ng ml spaced 100- to 300-mm apart and equidistantly. Using this technology, arrays can be constructed with more than 30,000 cDNAs on a single microscope slide 30 . The spotting is, wherever possible, carried out by a robot and the technology applied relies on a nib, piezo-, or ink-jet spotter. The cDNA-spotted arrays suffer slightly from at least two drawbacks first, the relatively large size of the probe sequence...

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