Abstract: The present invention provides a lysis buffer comprising a chaotropic agent and a reducing agent suitable for liquefaction of a highly viscous liquid biological sample, such as sputum, a use of said lysis buffer for the processing of a highly viscous liquid biological sample, methods for processing or analyzing a highly viscous liquid biological sample, or a method for detecting a pathogen within a highly viscous liquid biological sample. Furthermore, the present invention relates to a lysate comprising a highly viscous liquid biological sample and the lysis buffer according to the present invention, a ready-to-use reaction tube and a kit comprising the lysis buffer according to the present invention.

Abstract: Presented are methods of isolation of pili and pilus-like structures from Gram-positive bacteria including Streptococcus pneumoniae and compositions that include such isolated pili. These compositions are useful as immunogenic compositions for the production of antibodies and immunostimulation. Also presented are methods of inhibiting Streptococcus pneumoniae, and methods of identifying inhibitors of Streptococcus pneumoniae.

Abstract: A method and apparatus for extracting oil from algae comprises a disruptor (125) having a plurality of deflectors (220 and 225) against which a mixture of algae and water (100) is forcibly impacted upon the urging of a pump (115). The impacting of the algal cells against the deflectors ruptures their cell walls and liberates the lipids (oil) and other materials contained therein. A tank (130) collects the mixture and after a settling period, the mixture forms at least three layers comprising oil (155), water (160), and algal residue (165). The oil layer is removed through one or more conduits (177, 178) into a holding tank (185) for further refining and use. The water is discarded, and the biomass residue comprising algal cell walls and other non-oil components is removed to another holding container (199) from which it can be discarded or used as an agricultural fertilizer or the like. If desired, the residue can be further treated in order to scavenge any remaining oil.

Abstract: The present invention relates to, among other things, a device for collecting airborne microorganisms, said device having: an air collecting module, comprising: i. an upper element having an air admission duct permitting entry of an air stream into said module, said duct being provided, at its base, with means for disturbance of the air stream, ii. a lower element having air evacuating means permitting the air stream created to exit and said upper and lower elements can be made integral with one another so that the air stream can be created within said air collecting module; a cartridge, of roughly cylindrical shape, having a microorganism retention zone, said retention zone having means for lysis of the microorganisms, said cartridge being positioned within said air collecting module.

Abstract: The present invention is a bacteriolytic agent containing a cationic surfactant (A) of which a counteranion is an acid having a pKa (25° C.) of 0 to 10. As the counterion, a carboxylate anion is preferable. As the cationic surfactant (A), a quaternary ammonium salt-type surfactant is preferable. Examples of the useful substance include a protein, an amino acid, a nucleic acid, an antibiotic, sugars or vitamins. The bacteriolytic agent of the present invention is excellent in a bacteriolytic power in a step of extracting a useful substance from a microorganism (useful substance producing bacterium etc.). In addition, denaturation of the useful substance during the step is small.

Abstract: The inventive self-contained apparatus for isolating nucleic acid, cell lysates and cell suspensions from unprocessed samples apparatus, to be used with an instrument, comprises at least one input, and: (i) a macrofluidic component, comprising a chamber for receiving said unprocessed sample from a collection device and at least one filled liquid purification reagent storage reservoir; and (ii) a microfluidic component in communication with said macrofluidic component via at least one microfluidic element, said microfluidic component further comprising at least one nucleic acid purification matrix; and (iii) a drive mechanism on said instrument for driving said liquid purification reagent, through said microfluidic element and said nucleic acid purification matrix, wherein the only inputs to said apparatus are via said chamber and said drive mechanism.

Abstract: The invention relates to a method and a device for the digestion of biological cells. Digestion of cells is understood as breaking up cells so that hereditary information can be removed. Using the present invention, nucleic acids are to be able to be removed from cells in particular. The nucleic acids include RNA and DNA. According to the invention, cells are digested in a vessel which can be connected to a line and can thus be part of an overall device. Liquid can drain out of the container or can be pumped into the container via the line, for example. The typical means for performing a mechanical digestion are located in the vessel, that is, in particular beads and a buffer solution. In contrast to the prior art, however, the vessel is not shaken, that is, subjected to vibrations. Instead, it is stirred with the aid of a magnetic stirrer. Strong vibrations are thus avoided. A vessel for performing digestion of cells can therefore be integrated without problems in an overall device.

Abstract: The invention generally relates to methods and kits for isolating nucleic acids from an organism. In certain embodiments, methods of the invention involve contacting a plurality of lytic enzymes to an organism, thereby lysing a cell wall of the organism to release the nucleic acid, and introducing at least one agent to separate the nucleic acid from the lysed cells, thereby isolating the nucleic acid.

Abstract: The invention relates to a lysis buffer mixture that is stable in storage for isolating nucleic acids from biological, preferably diagnostic samples. The mixture is preferably associated with an extraction control. The aim of the invention is to provide an improved nucleic acid extraction system, which is cost-effective, stable and easy to use, thus fulfilling the requirements of a modern nucleic acid extraction system and containing, among other things, extraction controls. The invention relates to a lysis buffer mixture for isolating nucleic acids, said mixture containing non chaotropic salts, a special selection of detergents, a defined quantity of at least one nucleic acid as an extraction control, optionally lytic enzymes, optionally carrier nucleic acids and optionally other additives.

Abstract: The invention provides genes that are unique either to Ehrlichia ruminantium strain Gardel or to Ehrlichia ruminantium strain Welgevonden, or allelic couples which are present in both strains but whose sequences differ between the two strains, as genetic markers to differentiate between these two strains. The invention also provides diagnostic methods using these genetic markers.

Abstract: An analyte is separated from a fluid sample by introducing the sample into a cartridge having a sample port and a first flow path extending from the sample port. The first flow path includes an extraction chamber containing a solid support for capturing the analyte from the sample. The cartridge has a second flow path for eluting the captured analyte from the extraction chamber, the second flow diverging from the first flow path after passing through the extraction chamber. The sample is forced to flow through the extraction chamber and into a waste chamber, thereby capturing the analyte with the solid support as the sample flows through the extraction chamber. The captured analyte is then eluted from the extraction chamber by forcing an elution fluid to flow through the extraction chamber and along the second flow path.

Abstract: The invention provides a composition containing an aqueous liquid phase, bacterial cells, and an ionic compound dissolved in the liquid phase. The ionic compound is selected from the group consisting of 1-butyl-3-methyl-imidazolium-thiocyanate, 1-butyl-3-methyl-imidazolium-2(2-methoxy-ethoxy)ethylsulfate, 1-methyl-1-[4-(3-methyl-3H-imidazol-1-ium)-but-1-yl]-3H-imidazolium-di(toluylsulfate), and 1-butyl-3-methyl-imidazolium-octylsulfate. The compositions of the invention are advantageously used for preparing lysates of biological cells, particularly bacterial cells.

Abstract: Provided are surprisingly beneficial osmotic stress shock methods for facilitating disruption and/or extraction of microorganisms, comprising a synergistic combination of at least one hypotonic shock and at least two hypertonic shocks, wherein the last shock is a hypertonic shock. Preferred aspects of the invention relate to facilitating disruption of a broad spectrum of microorganisms (e.g., algae, bacteria, yeast, fungus, etc.). In particular aspects the microbial cells are subjected to: a hypertonic/hypotonic/hypertonic tertiary shock; a hypotonic/hypertonic/hypertonic tertiary shock; or a hypotonic/hypertonic/hypotonic/hypertonic quaternary shock. Particular aspects further comprise disrupting and/or extracting of the shocked microbial cells, and isolating a cellular constituent or bioproduct (e.g., biofuel, biocrude, bioenergy, biogas, biodiesel, bioethanol, biogasoline, pharmaceuticals, nutraceuticals, food, vitamins, feedstock, dyes, colorants, sulfur, fertilizer, bioplastic) therefrom.

Abstract: The present invention relates to the simple, gentle, and efficient extraction of biological material from Escherichia coli (E. coli). The use of E. coli in research laboratories depends on the ability to prepare lysates to isolate the desired products under investigation. The present invention includes methods and engineered E. coli strains that are capable of rapid controlled lysis or herein “autolysis”. The XJa strains were made from JM109 and the XJb strains from BL21 by insertion of the ? R or (? SR) lytic endolysin gene to replace the tightly regulated araB gene. Thus, arabinose becomes a non-metabolizable inducer and the controlled autolysis phenotype is induced by the PBAD promoter by the presence of saturating arabinose. Upon induction of the bacteriophage ?R endolysin, the E. coli remains intact but is efficiently lysed after one freeze-thaw cycle. The present invention is usable with many different buffer systems and is flexible in this regard.

Abstract: Methods for improving binding of a proteinaceous substance to cell-wall material of a Gram-positive bacterium are disclosed. The proteinaceous substance includes an AcmA cell-wall binding domain, homolog or functional derivative thereof. The method includes treating the cell-wall material with a solution capable of removing a cell-wall component such as a protein, lipoteichoic acid or carbohydrate from the cell-wall material and contacting the proteinaceous substance with the cell-wall material.

Abstract: We have performed separation of bacterial and cancer cells from peripheral human blood in microfabricated electronic chips by dielectrophoresis. The isolated cells were examined by staining the nuclei with fluorescent dye followed by laser induced fluorescence imaging. We have also released DNA and RNA from the isolated cells electronically and detected specific marker sequences by DNA amplification followed by electronic hybridization to immobilized capture probes. Efforts towards the construction of a “laboratory-on-a-chip” system are presented which involves the selection of DNA probes, dyes, reagents and prototyping of the fully integrated portable instrument.

Abstract: Presented herein are exemplary systems and methods for extracting lipids from a wet algal biomass. An exemplary method comprises lysing a wet algal biomass with an insoluble granular lysing agent to create a lysate, creating a lipid-rich phase in the lysate, and separating the lipid-rich phase from the lysate. An exemplary system comprises a lysing station for creating a lysate from a wet algal biomass and a separation station for creating and separating a lipid-rich phase from the lysate. According to further exemplary systems and methods, ultrasound may be used in place of or in addition to a lysing agent to lyse the wet algal biomass.

Abstract: The present invention provides a method and device for collecting treating and analysis of biological or chemical material by introducing a source material into a specimen container, transferring the source material to a processing device and thermally, chemically and/or mechanically treating the source material to alter at least one constitutive characteristic of the source material and to release or create a target material from the source material.

Abstract: The invention relates to flow-through devices for mixing fluids and uses thereof for mixing and for lysing cells to release biological compounds of interest. The invention particularly encompasses flow-through methods for mixing and chemical methods for the isolation and purification of plasmid DNA from cell culture.

Abstract: It is an object of the invention to provide a process for easily producing a glucan, which assures a high degree of safety and which does not generate an oxidation off-flavor, without using a hydroxide of an alkali metal such as sodium hydroxide. In a process of the present invention, a physically pulverized yeast is autolyzed with an intracellular enzyme thereof, in an electrolyzed alkaline water which is obtained by electrolyzing water.

Abstract: Stable compositions comprising a nucleic acid binding support material comprising a substrate with functional groups attached to the substrate; a lysing enzyme; a water dispersible matrix material; and a saccharide, methods of using the compositions, and products and devices which include the compositions are disclosed.

Abstract: The invention relates to bacterial ghost preparation using betapropiolactone for final inactivation of bacteria.

Abstract: The present invention provides systems, methods, and devices for using acoustic energy.

Abstract: The present invention relates to a cartridge which can be positioned inside an air collection means and receive a means for recovering nucleic acids, said cartridge being substantially cylindrical and comprising a microorganism retaining zone, said retaining zone comprising microorganism lysis means. The invention also relates to a device for collecting microorganisms contained in the air and a device for microorganism lysis.

Abstract: A microfluidic cartridge includes a capture portion capturing target material on surfaces of nonmagnetic particles; and a separating portion separating target molecules from the target material captured on the surfaces of the particles. The microfluidic cartridge is mounted on an adaptor that is rotated by a rotating unit in order to separate a fluid including the target molecules from the particles through centrifugal force.

Abstract: The present invention relates to compositions, apparatus and methods useful for concurrently performing multiple, high throughput, biological or chemical assays, using repeated arrays of probes. A combination of the invention comprises a surface, which comprises a plurality of test regions, at least two of which, and in a preferred embodiment, at least twenty of which, are substantially identical, wherein each of the test regions comprises an array of generic anchor molecules. The anchors are associated with bifunctional linker molecules, each containing a portion which is specific for at least one of the anchors and a portion which is a probe specific for a target of interest. The resulting array of probes is used to analyze the presence or test the activity of one or more target molecules which specifically interact with the probes. In a preferred embodiment, a sample to be tested is subjected to a nuclease protection procedure before it is contacted with a combination of the invention.

Abstract: Disclosed is a method for the dissociation of cells. Cells are processed under conditions of pH, temperature, and shear to thereby yield a mixture of cell wall ghosts and cytoplasm. Preferably, the cells are jet cooked at an alkaline pH to form an intermediate mixture, and the intermediate mixture is subsequently subjected to mechanical homogenization. Generally, the cells become dissociated, whereby at least one separate cell wall component is substantially separate from the dissociated cell walls.

Abstract: The present invention relates a method for the enumeration of mammalian cell micronuclei, while distinguishing micronuclei from the chromatin of dead and dying cells. The method utilizes differential staining of chromatin from dead and dying cells, to distinguish the chromatin from micronuclei and nuclei that can be detected based upon fluorescent emission and light scatter following exposure to an excitatory light source. Counting of micronuclei events relative to the number of nuclei can be used to assess the DNA-damaging potential of a chemical agent, the DNA-damaging potential of a physical agent, the effects of an agent which can modify endogenously-induced DNA damage, and the effects of an agent which can modify exogenously-induced DNA damage. Kits for practicing the invention are also disclosed.

Abstract: The present invention provides a plasmid pSOFLysK contained in the bacterial strain Lactococcus lactis NZ9800 referred herein as Lactococcus lactis NZ9800-pSOFLysK (subsequently designated Lactococcus lactis DPC6132) encoding anti-staphylococcal activity as deposited with DSMZ under accession No. ncimb 41409 and plasmids substantially similar thereto also providing anti-staphylococcal activity. In another aspect, the present invention provides a gene encoding an anti-staphylococcal protein, Lysin (LysK) as encoded by the plasmid pSOFLysK in Lactococcus lactis/VZ9800-pSOFLysK (subsequently designated Lactococcus lactis DPC6132). The recombinant lysine also has applications in diagnostics given its lytic mechanism.

Abstract: A method of treating a liquid sample having microbiological target species therein to concentrate the species and collect lysate is disclosed. The liquid sample comprises non-target microbiological particles, inorganic particles, and microbiological target species. The liquid is passed through a prefilter medium to allow the target species to pass through as filtrate and retain non-target microbiological products and inorganic particles thereon. The filtrate is contacted with a main filtration medium adapted to retain the target species thereon as retentate. The retentate is lysed to form a lysate containing target material that was enveloped within the microbiological target species. The microbiological species may comprise cell containing or viral material. Target materials comprise intracellular nucleic acids, or in the case of viral sampling, nucleic acids encased within the protein sheath or coating of the virus.

Abstract: Apparatus and method for collection of a target material from a liquid sample comprising target species that contain the target material. The apparatus comprises a fluid flow conduit in communication with a filter medium having a pore size adapted to retain target species thereon and pass, as filtrate, lysate containing the desired target material. A lysing agent conduit communicates with the fluid flow conduit and delivers lysing agent to the filter medium to lyse the target cells thereby releasing the desired intracellular target material. The lysing agent may be recirculated through the filter medium for a sufficient time to permit sufficient quantity of lysate to circulate through the system for lysate collection and subsequent assay.

Abstract: A device for sample tissue disruption and/or cell lysis comprising: a piezoelectric material; and at least a second material in contact with the piezoelectric material; and wherein the second material has an uneven surface on an opposite side to that in contact with the piezoelectric material. The device may be made by assembling at least three layers and membranes for the valves and pumps. The piezoelectric material is actuated by an external voltage source to generate cavitation, which disrupts tissue and/or lyses cells, in particular by a modulated alternative external voltage. The invention further provides a method of disrupting tissue and/or lysing cells in a device. Also provided is a piezoelectric device comprising a piezoelectric material in contact with a second material, and wherein the second material has an uneven surface on an opposite side to that in contact with the piezoelectric material.

Abstract: Provided are a microfluidic device including an electrolysis device for cell lysis which includes an anode chamber, a cathode chamber and a separator, in which the separator is installed between the anode chamber and the cathode chamber, the anode chamber includes an inlet and an outlet for an anode chamber solution and an electrode, and the cathode chamber includes an inlet and an outlet for a cathode chamber solution and an electrode, and a method of electrochemically lysing cells using the same.

Abstract: A method for the disruption of biological cells by means of a homogenizer device which a) comprises an orifice plate having at least one inlet nozzle and an orifice plate having at least one exit nozzle, wherein, in the intermediate space between the orifice plates, a static mixer is situated and, if appropriate, mechanical energy is additionally introduced or b) comprises an orifice plate having at least one inlet nozzle and a baffle plate, wherein, in the intermediate space between the orifice plate and the baffle plate, if appropriate a static mixer is situated and/or mechanical energy is introduced.

Abstract: The invention relates to methods and agents for thermal conditioning of a biological cell, methods and uses of said agents for releasing and insulating ingredients outside the biological cell, and for quantitatively and qualitatively detecting the released ingredient and to a method for sampling biological cells which consists in carrying out in one step the sampling said biological cells in a culture medium and in releasing cellular ingredients during said sampling. Sampling devices are also disclosed.

Abstract: Methods and apparatus for the selection or processing of particles sensitive to the application of an external stimulus, in which is produced, by applying the external stimulus, the rupture/lysis of at least one selected particle or the fusion of first and second selected particles, by means of the organisation of the particles using a first field of force by selectively energising electrodes of an array of selectable electrodes having dimensions comparable to or smaller than those of the particles, applying to the electrodes a first configuration of stresses; and by applying to the electrodes a second configuration of stresses, so as to create a second field of force, located substantially close to at least one selected particle to be lysated or to a pair of first and second particles to be fused and such as to produce the application of a stimulus suited to produce their lysis or fusion.

Abstract: Devices, processes, and kits for the extraction of nucleic acids from biological samples are disclosed. The devices comprise a first port, a second port, and a binding chamber intermediate and in fluid communication with the first port and the second port. The binding chamber comprises an unmodified flat glass surface effective for binding a heterogeneous population of nucleic acids. The first port, second port, and binding chamber define a continuous fluid pathway that is essentially free of nucleic acid-specific binding sites.

Abstract: An analyte is separated from a fluid sample by introducing the sample into a cartridge having a sample port and a first flow path extending from the sample port. The first flow path includes an extraction chamber containing a solid support for capturing the analyte from the sample. The cartridge has a second flow path for eluting the captured analyte from the extraction chamber, the second flow diverging from the first flow path after passing through the extraction chamber. The sample is forced to flow through the extraction chamber and into a waste chamber, thereby capturing the analyte with the solid support as the sample flows through the extraction chamber. The captured analyte is then eluted from the extraction chamber by forcing an elution fluid to flow through the extraction chamber and along the second flow path.

Abstract: The present invention provides a method of isolating nucleic acid from a sample of cells, said method comprising: (a) binding cells in said sample in a solid support to isolate cells from the sample; (b) lysing the isolated cells; and (c) binding nucleic acid released from said lysed cells to said same solid support and a kit for carrying out such a method. The method may advantageously be used to prepare nucleic acid for use in a nucleic acid-based target cell detection method.

Abstract: A differential extraction system is directed to a microfluidic-based integrated cartridge to automate differential extraction of specific cell types within a mixed sample. The integrated cartridge includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.

Abstract: A method for the production of the N-terminal four kringle-containing fragment of hepatocyte growth factor (NK4) by expression of a nucleic acid encoding said NK4 in a microbial host cell, isolation of inclusion bodies containing said NK4 in denatured form, solubilization of the inclusion bodies and naturation of the denatured NK4, characterized in that solubilization and naturation are performed at pH 7-9 in phosphate buffered solution, provides NK4 in high purity and high yield.

Abstract: The present invention provides a method of selectively lysing non-viable cells from a cell population within a sample, the method including selectively lysing non-viable cells by mixing a cell-containing sample with a lysis buffer including a non-ionic surfactant and a divalent cation salt.

Abstract: A sonication apparatus is directed to a microfluidic-based system to automate differential extraction of specific cell types within a mixed sample. The microfluidic-based system includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.

Abstract: Methods for improving binding of a proteinaceous substance to cell-wall material of a Gram-positive bacterium are disclosed. The proteinaceous substance includes an AcmA cell-wall binding domain, homolog or functional derivative thereof. The method includes treating the cell-wall material with a solution capable of removing a cell-wall component such as a protein, lipoteichoic acid or carbohydrate from the cell-wall material and contacting the proteinaceous substance with the cell-wall material.

Abstract: The present invention provides methods for the reduction of endotoxins in a plasmid preparation using a carbohydrate non-ionic detergent with silica chromatography.

Abstract: A process for treating a microorganism-containing stream includes the steps of passing the stream through a chamber, pressurizing the stream in the chamber to a pressure greater than 14.7 p.s.i.g., introducing a feed gas into the pressurized stream such that the feed gas is soluable within the microorganisms, and depressurizing the stream so as to cause the soluablized feed gas to expand within the microorganisms so as to rupture the cell walls of the microorganisms. The feed gas can be carbon dioxide, air, nitrogen, methane, or mixtures thereof.

Abstract: This invention describes a rapid (10 to 15 minutes), simple, flexible and efficient method of nucleic acids extraction for nucleic acid testing assays. This method has the following basic steps: i) mechanical cell lysis using solid particles in the presence of a chelating agent, followed by ii) controlling the presence and/or activity of NAT assays inhibitors. This method is applicable to various biological samples and universal for microorganisms, as one can use it to extract nucleic acids from test samples containing target viruses, bacteria, bacterial spores, fungi, parasites or other eukaryotic cells, including animal and human cells.

Abstract: Provided are a microfluidic device including an electrolysis device for cell lysis which includes an anode chamber, a cathode chamber and a separator, in which the separator is installed between the anode chamber and the cathode chamber, the anode chamber includes an inlet and an outlet for an anode chamber solution and an electrode, and the cathode chamber includes an inlet and an outlet for a cathode chamber solution and an electrode, and a method of electrochemically lysing cells using the same.

Abstract: A biological growth reactor vessel for the cultivation of micro-algae, diatoms or other unicellular organisms, especially for oil production, is described which incorporates a dispensing rod to which are attached a plurality of clear paddles which increase surface area for the diffusion of light and growth enhancing admixtures such as CO2, nitrogen and cellulose premixed at micron level. The dispensing rod incorporates a plurality of holes strategically placed to disperse the micron-mixture slurry of the growth admixtures. The dispensing rod and its attached clear paddles make use of methods and technologies to concurrently micron mix admixtures such as natural gases, amendments and other biological factors such as enzymes and microbes to increase contact area between said mixtures and micro-algae, diatoms or other naturally occurring unicellular growth.

Abstract: Apparatus and methods for detecting molecules in a fluidic environment are provided, including nanodevices and methods for fabricating, functionalizing, and operating such nanodevices. At least one of the methods includes selective heating of nanodevices in an array.