Cell disruption, biological

Ultrasound in Biology and Medicine Biological cell disruption... 

Most chemists working on sonochemistry in the laboratory will either use some form of ultrasonic bath or a commercial probe system. The latter instruments are often equipped with a pulse facility which was originally designed for biological cell disruption where temperature control is important. This pulse facility enables the power ultrasound to be delivered intermittently and thereby allow periods of cool-... 

Biological Methods Enzymatic digestion of the cell wall is a good example of biological cell disruption. It is an effective method that is also very selective and gentle, but its high cost makes it impractical to be used for large-scale operations. 

Originaiiy uitrasound probes were simpiy adaptations of biological cell disrupters. 

When the problem is to disrupt Ughtly bonded clusters or agglomerates, a new aspect of fine grinding enters. This may be iUustrated by the breakdown of pigments to incorporate them in liquid vehicles in the making of paints, and the disruption of biological cells to release soluble produces. Purees, food pastes, pulps, and the like are processed by this type of mill. Dispersion is also associated with the formation of emulsions which are basically two-fluid systems. Syrups, sauces, milk, ointments, creams, lotions, and asphalt and water-paint emulsions are in this categoiy. 

Biology, Biochemistry Homogenisation and cell disruption Power ultrasound is used to rupture cell walls in order to release contents for further studies. 

Maximum disruption is obtained in a zone close to the probe tip and the biological cells must be kept here for sufficient time to allow disruption to take place. A delicate balance must therefore be struck between the power of the probe and the disruption rate since power ultrasound, with its associated cavitational collapse energy and bulk heating effect, can denature the contents of the cell once released. Indeed for this type of usage it is important to keep the cell sample cool during sonication. The method is very effective and continues to be an important tool in microbiology and biochemistry research. 

Many biological cells contain degradative enzymes (proteases) that catalyze the hydrolysis of peptide linkages. In the intact cell, functional proteins are protected from these destructive enzymes because the enzymes are stored in cell organelles (lysosomes, etc.) and released only when needed. The proteases are freed upon cell disruption and immediately begin to catalyze the degradation of protein material. This detrimental action can be slowed by the addition of specific protease inhibitors such as phenylmethyl-sulfonyl fluoride or certain bioactive peptides. These inhibitors are to be used with extreme caution because they are potentially toxic. 

When carrying out cell disruption operations it is often necessary to provide cooling of the cell concentrate due to the high pressures developed in the equipment. An additional consequence of high-pressure operation is that cell disruption equipment can generate aerosols which may be undesirable, particularly for biologically hazardous organisms. In these cases, the ability to steam sterilize the equipment is required, for decontamination, and some type of secondary containment may also be required, such as an isolator or a contained area within a facility to which access is controlled. 

Chemical pretreatment of biological sludge using acid, base, or enzyme may enhance the sonication effect. Such chemicals weaken the cell wall, effectively disrupting the biological cells with less energy input. Bases such as NaOH and KOH are widely... 

The effects of US on surviving cells may include structural changes and interactions with deoxyribonucleic acid (DNA) [83]. The biological effects observed in vitro include fragmentation of cell membranes caused by the collapse of cavitation bubbles, microstreaming near the boundary layer and formation of radicals, which promote chemical reactions leading to wall decomposition [84]. Carstensen et al. [85] found the extent of cell disruption to be inversely proportional to the cell concentration. 

A tubular sonicatlon device was recently reported by Borthwick et al. [93] (see Fig. 3.9). The device requires the addition of no chemical, enzyme or particles that might complicate the subsequent determination step. Furthermore, denaturatlon of target DMA or proteins for detection Is minimized as the device tolerates moderate temperature rises this allows the use of sensitive and specific Immunological detection methods on sonicated biological materials. Because the tubular device Is composed of a piezoelectric resonator made of several material layers, selection of an appropriate operating frequency Is essential to ensure proper performance (i.e. acceptable cell disruption efficiency). This device can be used for batchwise treatment of small sample volumes or In flow systems without the risk of hazardous aerosol formation inherent in probe sonloators. 

High power ultrasound alone is known to damage or disrupt biological cell walls for the release of contents (vide supra). This will result in the destruction of the cells, i.e. sonication has the potential for the destruction of bacteria. Unfortunately very high intensities are required if ultrasound alone is to be used for complete sterilization. It has been shown however that low-power ultrasound is capable of... 

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