Abstract: The present invention relates to a 3-dimensional cell to tissue and maintenance process, more particularly to methods of culturing cells in a culture environment, either in space or in a gravity field, with minimum fluid shear stress, freedom for 3-dimensional spatial orientation of the suspended particles and localization of particles with differing or similar sedimentation properties in a similar spatial region.

Abstract: The process of the present invention relates to a three dimensional co-culture process. Wherein two distinct types of mammalian cells are co-cultured in a rotating bioreactor which is completely filled with culture media and cell attachment substrates. As the size of the tissue assemblies formed on the attachment substrates changes the rotation of the bioreactor is adjusted accordingly.

Abstract: An apparatus is disclosed for the controlled production of microorganisms by photosynthesis in a closed photobioreactor. The closed photobioreactor contain a photosynthetic culture in a substantially sealed environment and provides a system for recirculating the reactant gas through the culture. This closed loop system can be operated with expensive carbon isotopes (i.e., .sup.13 CO.sub.2 or .sup.14 CO.sub.2). Also, a system is provided for removing the molecular oxygen produced in the photosynthesis reaction from the closed photobioreactor. Furthermore, a pH-regulated control valve is utilized for controlling the addition of reactant gas to the culture in response to the alkalization of the culture.

Abstract: A unique hollow fiber bioreactor system and method for the propagation of cells and the production of various cell propagation products is described. The system and method include the use of a flow block holder which allows a dissolved oxygen probe to be calibrated at the same pressure as the media in the bioreactor system.

Abstract: An apparatus for controlling the fermentation of moromi mash comprises a sampling mechanism for sampling a portion of the prescribed amount of moromi mash stored in at least one storage tank, and an automatic multiple analyzer for receiving the portion of the prescribed amount of moromi mash from the sampling mechanism and simultaneously analyzing the concentrations of at least two ingredients of the sampled portion of the moromi mash.

Abstract: A multienzyme system capable of reducing waste products and synthesizing nutrients in a cell culture system is disclosed. In one embodiment, lactic dehydrogenase and leucine dehydrogenase are employed in conjunction with amino acid precursors and a recycling co-factor to achieve the synthesis of amino acids and a reduction in the levels of ammonia and lactic acid. The multienzyme system of the invention may be added directly to the cell culture system or it may be microencapsulated. Alternatively, the invention may be utilized in conjunction with a container unit in which cell culture medium contacts the multienzyme system external to the main culture vessel compartment.

Abstract: A flow-through cell cultivation apparatus and method is described. The flow-through cell cultivation system comprises a flowcell, a chamber inside the flowcell for holding cells, and a flowcell holder. The flowcell has a lower intake port for flowing liquid nutrient through a cellbed inside the flowcell and an upper outlet port for flowing the liquid nutrient out of the flowcell. The flowcell also includes a chuck for receipt of a thermal probe. The probe is made of electromagnetically non-interactive material. The flowcell is enclosed inside the flowcell holder. The flowcell holder includes a pair of intake ports into a cavity having an open end at the flowcell for turbulently flowing a temperature controlled gas against the flowcell. The cavity has rough walls to promote the turbulent flow. The flowcell holder includes an exhaust port for flowing the gas out of the flowcell holder which also serves as a port for another thermal probe.

Abstract: A temperature monitoring method and apparatus for monitoring the temperature within a mass of organic matter moved through a composting vessel. An elongated, stationary probe extends through the vessel from one end toward another end thereof. A plurality of temperature measuring devices are mounted along the probe. The probe may extend through a compaction ram and be provided with a sleeve for accommodating movement of the ram relative to the probe. A decoupling device provides for decoupling of the probe from a mounting base and an extraction device permits the probe to be extracted from the vessel for replacement thereof. The probe provides a method of monitoring temperature in the mass and a method of composting by monitoring temperature in the mass and regulating the temperature of the mass in response thereto.

Abstract: A method of treating humans suffering from central nervous system diseases, such as Alzheimer's disease, Parkinson's disease, senile dementia. The treatment consists of inducing into the patient's blood stream at least one from the group consisting of: sex hormones and anabolic hormones. Growth hormone is also be used in those patients in advanced stages of the disease, or in those patients where it has been determined that a low level of growth hormone is present. A method of diagnosing Alzheimer's disease, senile dementia, by the determination of the levels of the hormones somatotropin (human growth hormone) and somatomedin-C (IGF-I) after the administration of the Aroonsakul-Allen provocative test is also disclosed. Blood-sera samples are taken at certain time periods after the administration of L-Dopa, and the samples are tested for the levels of these hormones. These levels are then compared against the levels for normal subjects.

Abstract: A process for producing a physiologically active substance by a combined enzymatic method is disclosed.

Abstract: The cell culturing system of the present invention includes a tubular membrane, e.g. hollow fiber, cartridge. Provision is made for the continuous circulation of a nutrient-containing medium through the tube side of the cartridge by pumping the nutrient-containing medium in an endless loop. The cells to be cultured are placed within the cartridge on the shell side where the desired extracellular products are accumulated. Nutrients are assimilated into the cell culture through the semipermeable tubular membranes and waste products are removed by passing through the semipermeable membranes into the recirculating nutrient-containing medium. Within the endless loop are located a circulating pump, an oxygenator and a pH probe. The system additionally includes a variable delivery feed pump for continuously injecting fresh nutrient-containing solution into the loop, which feed pump is operated at a rate responsive to the measured pH.

Abstract: Method and apparatus for the determination of biomass in a suspension, such as a culture, in `real` time and a fermentation process using the method. In the method the electrical capacitance of the suspension is measured and from the measurement the volume fraction of the suspension enclosed by the cytoplasmic membranes of the cells (biovolume) is determined.

Abstract: To improve the yield and/or reduce the energy cost in carrying out a microbiological or enzymatic process in a reactor and to make the reaction conditions essentially independent of the size of the reactor, it is proposed to make use, as a reactor, of an endless circulation tube in which the reaction components are circulated essentially according to a plug flow and in this process are fed through one or more in-line mixers fitted inside the tube. This method and reactor are suitable in particular for the preparation by fermentation of polysaccharides, especially xanthan, in which water, a production medium containing one or more sugars and nutrient salts and an inoculating material of a suitable aerobic bacterium are introduced into the said reactor tube and exposed to fermentation with air being supplied.

Abstract: Exoenzymes, such as proteases, xylanases and amylases, are obtained continuously by cultivation of exoenzyme-producing microorganisms in one step in a fermenter which is operated with continuous flow and in which a deficiency state corresponding to maximal enzyme productivity is effected. Optical density of the culture (as a measure of the biomass density) and exoenzyme concentration in culture can be monitored to control the timing and extent of the deficiency state. It is particularly advantageous to impose an oxygen limitation and to maintain the deficiency state continuously by exerting an effect on the oxygen input.

Abstract: Monoclonal antibodies to angiotensin II and the continuous hybrid monoclonal cell lines for their production are provided. These antibodies are useful in the diagnosis and treatment of angiotensin II-induced hypertension.

Abstract: A method of continuous product formation using at least two continuous fermentation units and a microorganism capable of being induced, in response to environmental conditions, to undergo a genetic alteration from a state favoring microorganism growth to a state favoring product production by the microorganism. The first continuous fermentation unit is maintained at environmental conditions selected to favor growth of the microorganism and to be nonpermissive for the genetic alteration. The microorganism is grown continuously in the first unit, and a portion of the growing microorganism cell mass is transferred via connecting means to the second continuous fermentation unit. Either the connecting means or the second unit is maintained at second environmental conditions selected to effect the genetic alteration. The altered microorganism is cultured in the second unit.

Abstract: A process and apparatus for measuring the degree of pollution, the degradableness, and the level of toxicity of aqueous liquids, for example, effluent. A partial stream is withdrawn from the liquid undergoing examination and diluted with biologically neutral water. The diluted partial stream is oxygenated and continuously flows through a biological bath in a reaction vessel having a constant living mass. The dilution of the partial stream is regulated in such a manner through measurement of the oxygen content upstream and downstream of the reaction vessel, that with constant volume flow through the reaction vessel, the difference between the measurement values obtained remains essentially constant at a predetermined value, so that the amount of dilution serves for the indication of the level of pollution. Additional reaction vessels connected in parallel or in series with the first reaction vessel and having associated oxygen electrodes allows measurement of the toxicity and degradableness of the pollution.

Abstract: A bioreactor and method of its operation are described comprising a vertical column with a lower reaction section filled with column packing particles classifiable by means of their differential settling rates in a liquid, and an upper disengagement section, preferably of larger diameter and of equal or greater volume than the reaction section, the two sections desirably being connected by a transition section in the shape of an inverted, truncated cone. Following initial classification of the packing in the reaction section, the packing is inoculated with microorganisms, and the bioreaction commenced by introducing nutrient feed at one end of the column, and withdrawing product at the other end. The packing is periodically cleaned, as required, by dispersing it into the disengagement section by passing liquid at an increased velocity upward through the column until contaminants have been dislodged.

Abstract: A method of controlling culture wherein cells are aerobically cultured to produce metabolite is characterized by adding substrate and/or inducer to the culture broth, judging the endpoint of the growth of the cells and/or the production of the metabolite or terminating the culture under the guidance of a change in the ratio of the first parameter based on the amount of carbon dioxide evolved by the cell culture to the second parameter based on the amount of oxygen consumed by the cell culture. The ratio is a respiratory quotient, for instance.

Abstract: A process for converting AMP into ATP which comprises (a) using an enzyme which converts AMP into ADP and has been produced from microorganisms having an optimum growth temperature of 50.degree. C. to 85.degree. C. and an enzyme which converts ADP into ATP and has been produced from microorganisms having an optimum growth temperature of 50.degree. to 85.degree. C. is disclosed. In addition, there is disclosed a process for producing a physiologically active substance by a multienzyme process which comprises forming ATP from AMP by the step (a), (b) synthesizing a physiologically active substance with the resulting ATP, coverting AMP resulting from the reaction in step (b) into ATP by the reaction in step (a), and repeatedly utilizing the converted ATP for synthesis of the physiologically active substance in step (b). By using the process it is possible to stably and efficiently carry out conversion of AMP into ATP over a long period of time.

Abstract: This invention relates to the production of the microorganisms. The operation is conducted in two stages. The first stage is conducted in the absence of ultrafiltration while the pH of the growth medium is maintained within the range of 6-7 by the addition of a neutralizing agent. Nutrient substratum and dilution water are added during the first stage to maintain the growth rate at a constant level, and the volume of growth medium is permitted to increase. When the amount of growth inhibiting agent produced in the fermentation reaches a predetermined maximum level, the second stage is initiated and the growth rate is maintained in a range of 0.10 to 0.50/hr, which is less than the growth rate in the first stage. By increasing the amount of dilution water the concentration of the growth inhibiting agent is maintained so as to achieve the desired moderate growth rate.

Abstract: A method and apparatus for the continued incubation of cells in a liquid culture medium. When the pH value of the culture medium deviates during the incubation from a pH range suited for the growth or multiplication of the cells, the culture medium is automatically replaced. Whether or not the pH value of the culture medium has deviated from the acceptable pH range is determined by measuring the intensity of light absorbed by the culture medium.

Abstract: This invention relates to a device constituted by a mixture of phyllosilicate and glucose powder, in which is embedded a filament connected to a source of alternating current. The mixture is contained in an inert envelope, the whole being contained in a container made of any material also containing the mixture and in which a test tube containing the culture medium is introduced.

Abstract: A process for the production of microbial cells by fermentation of carbonaceous material in a foam fermenter containing an oxygen-enriched nutrient medium. The process uses a source of carbon which is assimilable by the microorganism for the production of the microbial cells. The microbial cells are separated and removed from the foam fermenter for use as a food product high in protein content. The process includes the controlled release of a quantity of the constituents of a portion of the microorganism within the fermenter to increase the maintenance of the source of carbon and the nutrient medium in a foamed condition at a predetermined level in the fermenter. Also disclosed are various forms of apparatus for practicing the process of the present invention.

Abstract: A process and apparatus for measuring the degree of pollution, the degradableness, and the level of toxicity of aqueous liquids such as effluent. A partial stream is withdrawn from the liquid undergoing examination and diluted with biologically neutral water. The diluted partial stream is oxygenated and continuously flows through a biological bath in a reaction vessel having a constant living mass. The dilution of the partial stream is regulated in such a manner through measurement of the oxygen content upstream and downstream of the reaction vessel, that with constant volume flow through the reaction vessel, the difference between the measurement values obtained remains essentially constant at a predetermined value, so that the amount of dilution serves for the indication of the level of pollution. Additional reaction vessels connected in parallel or in series with the first reaction vessel and having associated oxygen electrodes allows measurement of the toxicity and degradableness of the pollution.

Abstract: Polysaccharides, such as xanthan gum, are produced by culturing microorganisms, e.g. of the Xanthomonas genus, in a two stage process. In the first stage, growth of the microorganism is favored, e.g. by using a predetermined quantity of a carbon-containing nutrient which does not support biosynthesis of the polysaccharide. In the second stage, the conditions are such that biosynthesis of the polysaccharide takes place with substantially no growth of the microorganism, e.g. by adding carbohydrate in the absence of nutrient required for polysaccharide growth.By this process, the requirement for oxygen is greatly reduced at the time when the culture medium has its highest viscosity, thereby minimizing problems of low oxygen transfer capability in viscous media.

Abstract: A method for production of HA fraction containing protective antigens of Bordetella pertussis (i.e. a fraction containing F-HA and LPF-HA) in an industrial scale, for instance, with a fermentater, which comprises inoculating a strain of B. pertussis in a liquid culture containing a cyclodextrin or its derivative, culturing it by a spinner culture under controlling the culture temperature and the amount of dissolved oxygen and under defoaming condition, optionally under controlling the pH range, and harvesting the produced HA fraction from the culture broth at a stage of from logarithmic growth phase to static grow phase, and a method for production of a pertussis vaccine from the HA fraction thus obtained by formalinizing the HA fraction in the presence of an amino acid.

Abstract: Troublesome foaming in the fermentation of yeast is suppressed by the addition of polyglycol ethers which are derived from saturated and monounsaturated C.sub.16 -C.sub.18 fatty alcohols and which contain on an average of from 1 to 4 ethylene and/or propylene glycol ether groups. Preferred defoamers contain oleyl and cetyl alcohol residues in a ratio of from 3:1 to 1:1, and on average 2 ethylene glycol ether groups. The defoamers do not inhibit yeast growth or lead to premature softening and liquefaction of the compressed yeast.

Abstract: The invention relates to a process for fermenting carbohydrate- and phosphate-containing liquid media with limitation of phosphate with bacteria capable of forming butanol, acetone and/or ethanol as fermentation products performed with a total phosphate content in the medium of 1.0 to 0.4 mmoles.

Abstract: Methods and devices for assaying the sensitivity of of biopsied cells to therapeutic agents are disclosed. Cells are cultured in artificial organs and then contacted with a fluorogenic substrate such that living cells accumulate a characteristic amount of fluorescence. The agent is then introduced into the organ and changes in the fluorescence released by the cells serve as an indicator of the sensitivity of the cells to the agent.

Abstract: A composition is used as a reagent in a method for measuring Antistreptolysin O (ASO) in a blood sample. The composition contains Streptolysin O (SO) at a pH outside the range of its hemolytic activity, e.g., outside the pH range of 5-9. At that pH the SO is reversibly inactivated while maintaining its hemolytic capacity. The composition is employed to measure ASO by mixing one or more samples of non-hemolyzed blood or serum with one or more known quantities of the composition with a known hemolytic capacity, incubating each mixture to allow reaction of any ASO in the sample with the SO, restoring the hemolytic activity of the SO by adjustment of pH to permit lysis and detecting the presence or absence of lysis in each sample.

Abstract: In a cell culture process, the addition of an additive is only carried out after it has been determined that the previous addition of additive has caused at least a minimum change in metabolic activity of the cells.

Abstract: A sterilizable tissue squeezer is disclosed, comprising a forcing frame fixed to selectively vertically displaceable relative to the lid of a sterile culture vessel. The forcing frame has at its lower end a pair of tissue interface members which may be gradually and accurately adjusted toward and away from each other to provide a direct mechanical force to living tissue growing in sterile culture for manipulating the plane of cell division therein to afford possible control of the emergence of form and promote establishment of stems and roots in culture.

Abstract: A process of calibrating a blood sugar analyzing apparatus, in which the blood sugar concentration in a blood specimen is measured with a fixed enzyme membrane sensor and corrected by calibration means, said blood sugar analyzing apparatus providing a linear relationship between measured and actual blood sugar concentrations in a range of blood sugar concentrations which are lower than a predetermined blood sugar concentration, and producing a deviation from said linear relationship in a range of blood sugar concentrations higher than said predetermined blood sugar concentration, in that said process comprises the steps of effecting a plurality of measurements of standard solutions having known low blood sugar concentrations in said blood sugar analyzing apparatus, obtaining an average value of the results of the measurements with said calibration means, determining a first correction coefficient (k.sub.

Abstract: A modular instrumentation system for monitoring and control of biochemical processes, and in particular of fermentation processes is provided. The system includes a plurality of function monitoring and control modules each including a microprocessor and associated memory devices, manual input devices and an interface for the receipt of sensor signals and the transmission of control signals. The modules for a plurality of functions have substantially common design and are adapted for relatively quick conversion to another function. The system may include an instrument console adapted to receive a plurality of the function monitoring and control modules as well as incorporating provision for sensor inputs, power inputs, one or more recorders, one or more pumps and/or an interface for an external computer.

Abstract: A method and apparatus are disclosed for determining the concentration of coliform bacteria in a sample. The sample containing the coliform bacteria is cultured in a liquid growth medium. The cultured bacteria produce hydrogen and the hydrogen is vented to a second cell containing a buffer solution in which the hydrogen dissolves. By measuring the potential change in the buffer solution caused by the hydrogen, as a function of time, the initial concentration of bacteria in the sample is determined. Alternatively, the potential change in the buffer solution can be compared with the potential change in the liquid growth medium to verify that the potential change in the liquid growth medium is produced primarily by the hydrogen gas produced by the coliform bacteria.

Abstract: A bioluminescent assay for ATP in water borne bacteria is made by adding nitric acid to a water sample with concentrated bacteria to rupture the bacterial cells. The sample is diluted with sterile, deionized water, then mixed with a luciferase-luciferin mixture and the resulting light output of the bioluminescent reaction is measured and correlated with bacteria present. A standard and a blank also are processed so that the light output can be correlated to bacteria in the sample and system "noise" can be substracted from the readings.A chemiluminescent assay for iron porphyrins in water borne bacteria is made by adding luminol reagent to a water sample with concentrated bacteria and measuring the resulting light output of the chemiluminescent reaction. The light output is correlated with bacteria present. A standard and a blank are also processed so that the light output can be correlated to bacteria in the sample and system "noise" can be subtracted from the readings.

Abstract: A process for aerobic composting or drying of organic waste materials in a composting bin and apparatus for carrying out the process. The composting bin is formed by one or more elongate composting passageway chambers which can be aerated or vented and heated. Each chamber encloses in sequence an intake zone, shaping zone, reaction zone, and discharge or transfer zone. Material is fed loosely through an opening into the intake zone in successive substantially equal portions or charges. A clearing element at the intake end of the chamber intermittently pushes each successive portion through the shaping zone, against the preceding portion, and through the reaction zone of the chamber. Residence time in the compost bin is sufficient for complete composting or other purposes before discharge or transfer. In the case of a bin with two or more chambers the chambers are arranged at different levels one below the other.

Abstract: Apparatus for preparation and distillation of low-alcohol-content, fermentation products into high-alcohol-content, fuel-grade product using solar energy to carry out the fermentation and distillation. The apparatus includes a solar collector with reflectors, fermenting tanks, a distillation column, and temperature controls. The working fluid for the solar collector is isolated from the fluid being distilled.

Abstract: Liquid (13, 13a) within a vessel (12, 12a) which receives an inflow is maintained at a predetermined level (43) by discharging liquid as necessary to compensate for the inflow. Discharge flow is drawn from below the surface (18) of the liquid to avoid clogging from scum or from other undesirable effects of surface draw off. Intake structure (34) for the discharge pump (33, 33a) has a branched flow path (47) that includes a first inlet (48) situated above the predetermined level, a second inlet (49) situated below the predetermined level and a flow junction (51) through which both inlets are communicated with the discharge pump, the junction being at the predetermined liquid level. If the liquid rises above the flow junction, the pump draws liquid through the second, subsurface inlet. If the liquid surface recedes below the flow junction, the pump aspirates air or other gas through the first inlet. Consequently liquid level is stabilized at the elevation of the flow junction.

Abstract: To establish predetermined humidity levels within a sterile treatment chamber, and maintain sterility, water, preferably deionized, is conducted to an evaporator outside of a container defining the treatment chamber, to be there heated to superheated steam, for example in the order of 300.degree. C., and conducted to a bypass duct (1) in atmospheric communication with the treatment chamber (10), for mixing with the atmosphere within the treatment chamber. Preferably, the treatment chamber is surrounded by heat exchanger elements, such as electrical heating wires and/or cooling coils, the heaters being controlled to rapidly heat the treatment chamber to a temperature in the order of 180.degree. C. during pauses of treatment for sterilizing the interior of the container, and/or maintaining temperature levels at a desired treatment level in operation.

Abstract: New and improved methodology for the diagnosis of a disease from blood sample analysis is disclosed and comprises: the preparation of a putative antigenic body substance extract, which is specific to disease-sensitized blood leukocytes, from the pooled, like body substances of a plurality of donors known to have the disease of interest; the mixture of said extract with the blood sample leukocytes; the promotion of the reaction therebetween to modify a characteristic of the disease-sensitized leukocytes, if any, of the blood sample; and the determination of the extent, if any, to which said characteristic has been modified. In multiple sclerosis diagnosis, said body substance may be blood or urine; while for malignant disease (for example breast cancer, or cancer of the head and neck) said body substance may be blood, urine, pleural fluid, ascites fluid, supernates of tumor cells or tumor cells grown in culture media.

Abstract: Apparatus for controlling the redox potential of a perfusate for use in the preservation of organs to be transplanted and the like, which apparatus includes first and second cells, a membrane with a submicron pore size disposed in openings between the cells, first and second electrodes disposed in the first and second cells respectively, a reference electrode adapted to be in electrical contact with the perfusate, and circuitry for detecting the redox potential of the perfusate as measured against the reference electrode and for maintaining the redox potential at a predetermined level by causing current to flow between the first and second electrodes through the perfusate when the measured redox potential differs from the predetermined level. The first cell has first and second ports for ingress and egress of the perfusate into and out of the first cell.

Abstract: This is an improved method for producing activated prothrombin complex concentrate which comprises controlling the degree of activation by determining the activation state of the starting material and then varying at least one of the activation conditions in accordance with analyses of the progress of activation of the starting material to arrive at a predetermined activation level.

Abstract: An automated micro-organism culture growth and detection apparatus includes a plurality of carousel trays arranged in pairs, providing receptacles for a plurality of culture bottles. The bottles each include a pair of electrodes submerged in the culture liquid and extend through an end wall of the bottle to form contact points. The receptacles include a pair of spring finger electrical contacts for each bottle to connect the electrodes to a measuring circuit. The complementary sets of the pairs of carousel trays are driven in counter-rotational directions to provide agitation for the culture during incubation and to index the individual to an access door in the incubation chamber surrounding the assembled carousel trays. A continuous flow of air at a controlled temperature is caused to flow through the incubation chamber which, together with the agitation, promotes the rapid growth of micro-organism in the culture bottles.

Abstract: The rate of enzyme reaction is determined by determining of the quantity of enzyme, and deriving the rate of reaction per unit time of a reaction solution having an absorbance which is proportional to the time. A temperature correction coefficient which is used during the determination of the enzyme quantity is derived by temperature information which is supplied by a temperature sensor disposed within the reaction solution to be determined.

Abstract: Method and automated apparatus are disclosed for determining the time of detection of metabolically produced hydrogen by coliform bacteria cultured in an electroanalytical cell from the time the cell is inoculated with the bacteria. The detection time data provides bacteria concentration values. The apparatus is sequenced and controlled by a digital computer to discharge a spent sample, clean and sterilize the culture cell, provide a bacteria nutrient into the cell, control the temperature of the nutrient, inoculate the nutrient with a bacteria sample, measures the electrical potential difference produced by the cell and measures the time of detection from inoculation.

Abstract: A fermentation vessel is used to grow a cell mass and maintain it in a maintenance energy state. In this state the mass of the cell material is maintained substantially constant, and fresh medium containing an energy source is introduced to the vessel at a rate sufficient to sustain cell viability but insufficient to support growth of the cell mass. Fluid with suspended cells is pumped from the vessel to a separator which separates a fraction of the fluid from the cells and discharges the fraction, the unfiltered portion of the fluid and substantially all of the cells being returned to the vessel, the discharged fraction containing useful secondary metabolites.