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Microbial content of nonsterile therapeutic agents containing natural of seminatural active ingredients.

The relationship was investigated between various chemical or pharmaceutical production processes and the extent of microbial contamination, of natural origin, of the resulting products. The products contained active ingredients of vegetable, enzymatic, or animal origin. It was concluded that (i) vegetable products practically free from microbes can be produced if the proper manufacturing steps are taken; (ii) sterilization of the media used to manufacture antibiotics, etc., produces products with little contamination; and (iii) products containing extracts of animal organs require careful refrigeration and addition of preservatives to produce acceptable levels of microbial contamination.

Bacteria↗

[Optimization of isolation of inulin from Cichorium intybus L. and some of its uses in social practice].

The study is concerned with the technological processing of chicory Cichorium intybus in order to isolate the fructan polysaccharide inulin (C6H10O5) and the optimization of the individual production steps of extraction (the method of purification of the raw material prior to the extraction itself and purification of the final product. temperature of the extraction mixture, exposure period of the extraction agent, effect of pH, optimization of the module for the determination of the most suitable volume of the extraction agent, determination of the most suitable ratio of the volume of ethanol in the extraction of inulin from the reaction medium). The obtained experimental results are compared and evaluated with regard to quality (whiteness) and the yield of the obtained final product. The paper is also concerned with several possible uses of inulin in pharmaceutical, medical, cosmetological, food industrial, and fodder production fields.

Chemistry, Pharmaceutical↗

Nisin and pediocin production on mussel-processing waste supplemented with glucose and five nitrogen sources.

AIMS: Optimization of bacteriocin production by L. lactis subsp. lactis CECT 539 and Ped. acidilactici NRRL B-5627 on mussel-processing wastes. METHODS AND RESULTS: The concentrations of glucose and five nitrogen sources that optimize nisin and pediocin production were determined using a second order orthogonal factorial design. NH4Cl, glycine and glutamic acid were poor nitrogen sources. Enhanced pediocin and nisin productions were achieved in a medium supplemented with yeast extract or Bacto casitone. CONCLUSIONS: Mussel-processing wastes could be successfully used as a culture medium for bacteriocin production at low production cost. SIGNIFICANCE AND IMPACT OF THE STUDY: Average optimization led to a threefold increase in pediocin production (from 322 to 934 BU ml(-1)) and in nisin production (from 32 to 100 BU ml(-1)) when compared with the unsupplemented medium. For this reason, mussel-processing waste warrants further investigation due to its potential use as a cheap culture medium for upscaling bacteriocin production.

Animals↗

The effect of home processing on postharvest fungicide residues in citrus fruit: residues of imazalil, 2-phenylphenol and thiabendazole in 'home-made' marmalade, prepared from late Valencia oranges.

The effects of domestic marmalade-making procedures on residues of imazalil, 2-phenylphenol and thiabendazole in 'sweet' oranges were investigated by comparing residue levels in raw fruit and in marmalades, prepared by heating the oranges with water and sugar for about 4 h in a preserving pan on a gas ring or for about 1 h in a microwave oven. Determination of residues in raw oranges and in marmalades was carried out by extraction with dichloromethane, followed by clean-up and reversed phase HPLC with UV detection for imazalil and fluorescence detection for 2-phenylphenol and thiabendazole. Residue analysis showed that the extent of carry-through of imazalil, 2-phenylphenol and thiabendazole residues into the marmalades depended on the method of marmalade-making and the nature of the fungicide. Persistence of fungicides was higher in the marmalade prepared in the microwave oven than in that prepared in the preserving pan. An increase (about 50%) in levels of imazalil in the fruit portions of marmalades compared with raw fruit residue levels indicated that imazalil was more readily extracted from oranges after processing. Comparison of 2-phenylphenol levels in raw fruit and fruit portions of marmalades showed that the longer exposure to heat, needed for marmalade-making in a preserving pan, reduced residue levels more severely (48%) than cooking in the microwave oven (13%). Thiabendazole levels in the fruit portion of marmalade prepared in the microwave oven stayed the same as in the raw fruit, but cooking in the preserving pan decreased residue levels by about 22%.

Biphenyl Compounds↗

Using electronic odor sensors to discriminate among oak barrel toasting levels.

Toasting changes both the quantity and the quality of the extractable substances in oak wood of barrels used for the aging of fine wines and spirits. Mastery and repeatability of toasting are vital in the production of quality barrels to be used for aging wines and spirits. Toasted wood components, which can be extracted by the wines or spirits during the aging process, are normally analyzed by maceration in standard alcohol solutions at concentrations adapted to the various products and can be used to control the intensity of the wood toasting. These kinds of analyses are accurate but time-consuming and need specialized laboratories. In this work, the feasibility of monitoring barrel toasting levels using an electronic nose with a metal oxide odor sensor array (MOS) was studied. The results of oak toasting level differentiation obtained via the MOS network were identical to those obtained by analyzing extractable compounds in liquid or gas phase as described in a previous paper. The results presented in this work at the laboratory scale could be used to implement a nondestructive monitoring system based on the analysis of headspace of barrels under industrial conditions.

Chemistry Techniques, Analytical↗

Characterization of surface runoff in urban areas.

Water quality measurements of surface runoff have been carried out in selected residential and industrial zones in urban areas, in which yearly mean precipitation is 1,225 mm. The concentrations of constituents in the surface runoff were measured at sampling sites categorized by land use type in the residential zone, and by industry type in the industrial zone. The water quality constituents of BOD5, COD, SS, NO3-N, TKN, PO4-P, TP, n-Hexane extracts, Cr, Cu, Pb and Fe were analyzed. The event mean concentrations (EMCs) of COD, SS, TKN and TP in the residential zone were 313 mg/L, 279 mg/L, 8.45 mg/L, 1.98 mg/L, and those in the industrial zone were 80 mg/L, 106 mg/L, 5.07 mg/L, and 1.93 mg/L, respectively. Cumulative load curves were created to analyze the first-flushing effect of each pollutant related to the pollutant, the rainfall event, and the land use type. No general relationship between the cumulative load and runoff has been established. The degree of first-flushing effect by constituents was in the following order; TKN>COD>SS>HEM>TP>PO4-P. The correlations between SS and other constituents were analyzed to evaluate the efficiency of the physical treatment process to control the surface runoff in urban areas. Based on the correlation of constituents with SS, high treatment efficiency of SS, heavy metals, organic matter, and TP was expected. The unit pollutant loading rates of COD, SS, TKN, TP, Cr and Pb in the residential zone were 2,392, 2,130, 64.6, 15.1, 0.31, and 1.83 kg/ha/yr, and those in the industrial zone were 612, 812, 38.7, 14.8, 0.51 and 0.82 kg/ha/yr, respectively.

Cities↗

Polymerase chain reaction-restriction fragment length polymorphism analysis of mitochondrial cytb gene from species of dairy interest.

Species identification plays an important role in food allergy prevention and food substitution detection that can reduce the commercial value of a product. For these reasons, many molecular methods have been developed to determine species origin; among them, polymerase chain reaction (PCR)-based methods were successfully applied to processed or unprocessed foodstuffs. An updated PCR-RFLP (restriction fragment length polymorphism) method of the cytb gene was developed for the identification of the 4 species of main interest in the dairy industry (Bos, Ovis, Capra, Bubalus). The comparative analysis of the 92 cytb sequences available in the database belonging to the 4 species allowed identification of 2 highly conserved regions, which were used to design 2 oligonucleotides for the PCR amplification of a 275 base-pair (bp) cytb fragment. The in silico analysis allowed identification of a set of species-specific restriction endonucleases (HaeIII, TaqI, and MwoI), which generated easily analyzable species-specific restriction profiles of the 275 bp cytb DNA fragment. The system was developed for both purified DNA and DNA extracted from meat or dairy products and finally tested on mixed samples, indicating its applicability to foodstuffs.

Animals↗

Mining of biological data I: identifying discriminating features via mean hypothesis testing.

Large volumes of data are routinely collected during bioprocess operations and, more recently, in basic biological research using genomics-based technologies. While these data often lack sufficient detail to be used for mechanism identification, it is possible that the underlying mechanisms affecting cell phenotype or process outcome are reflected as specific patterns in the overall or temporal sensor logs. This raises the possibility of identifying outcome-specific fingerprints that can be used for process or phenotype classification and the identification of discriminating characteristics, such as specific genes or process variables. The aim of this work is to provide a systematic approach to identifying and modeling patterns in historical records and using this information for process classification. This approach differs from others in that emphasis is placed on analyzing the data structure first and thereby extracting potentially relevant features prior to model creation. The initial step in this overall approach is to first identify the discriminating features of the relevant measurements and time windows, which can then be subsequently used to discriminate among different classes of process behavior. This is achieved via a mean hypothesis testing algorithm. Next, the homogeneity of the multivariate data in each class is explored via a novel cluster analysis technique called PC1 Time Series Clustering to ensure that the data subsets used accurately reflect the variability displayed in the historical records. This will be the topic of the second paper in this series. We present here the method for identifying discriminating features in data via mean hypothesis testing along with results from the analysis of case studies from industrial fermentations

Algorithms↗

A molecular beacon assay for measuring base excision repair activities.

The base excision repair (BER) pathway plays a key role in protecting the genome from endogenous DNA damage. Current methods to measure BER activities are indirect and cumbersome. Here, we introduce a direct method to assay DNA excision repair that is suitable for automation and industrial use, based on the fluorescence quenching mechanism of molecular beacons. We designed a single-stranded DNA oligonucleotide labelled with a 5'-fluorescein (F) and a 3'-Dabcyl (D) in which the fluorophore, F, is held in close proximity to the quencher, D, by the stem-loop structure design of the oligonucleotide. Following removal of the modified base or incision of the oligonucleotide, the fluorophore is separated from the quencher and fluorescence can be detected as a function of time. Several modified beacons have been used to validate the assay on both cell-free extracts and purified proteins. We have further developed the method to analyze BER in cultured cells. As described, the molecular beacon-based assay can be applied to all DNA modifications processed by DNA excision/incision repair pathways. Possible applications of the assay are discussed, including high-throughput real-time DNA repair measurements both in vitro and in living cells.

Animals↗

Protein purification using magnetic adsorbent particles.

The application of functionalised magnetic adsorbent particles in combination with magnetic separation techniques has received considerable attention in recent years. The magnetically responsive nature of such adsorbent particles permits their selective manipulation and separation in the presence of other suspended solids. Thus, it becomes possible to magnetically separate selected target species directly out of crude biological process liquors (e.g. fermentation broths, cell disruptates, plasma, milk, whey and plant extracts) simply by binding them on magnetic adsorbents before application of a magnetic field. By using magnetic separation in this way, the several stages of sample pretreatment (especially centrifugation, filtration and membrane separation) that are normally necessary to condition an extract before its application on packed bed chromatography columns, may be eliminated. Magnetic separations are fast, gentle, scaleable, easily automated, can achieve separations that would be impossible or impractical to achieve by other techniques, and have demonstrated credibility in a wide range of disciplines, including minerals processing, wastewater treatment, molecular biology, cell sorting and clinical diagnostics. However, despite the highly attractive qualities of magnetic methods on a process scale, with the exception of wastewater treatment, few attempts to scale up magnetic operations in biotechnology have been reported thus far. The purpose of this review is to summarise the current state of development of protein separation using magnetic adsorbent particles and identify the obstacles that must be overcome if protein purification with magnetic adsorbent particles is to find its way into industrial practice.

Adsorption↗

The promise of a virtual lab in drug discovery.

To date, the life sciences 'omics' revolution has not lived up to the expectation of boosting the drug discovery process. The major obstacle is dealing with the volume and diversity of data generated. An enhanced-science (e-science) approach based on remote collaboration, reuse of data and methods, and supported by a virtual laboratory (VL) environment promises to get the drug discovery process afloat. The creation, use and preservation of information in formalized knowledge spaces is essential to the e-science approach. VLs include Grid computation and data communication as well as generic and domain-specific tools and methods for information management, knowledge extraction and data analysis. Problem-solving environments (PSEs) are the domain-specific experimental environments of VLs. Thus, VL-PSEs can support virtual organizations, based on the changing partnerships characteristic of successful drug discovery enterprises.

Computer Simulation↗

Toxicity and hazardous properties of solvent base adhesive wastes.

In this work, the hazardous properties of solvent base adhesive wastes generated in the footwear manufacturing process have been studied. The characterisation procedures and criteria used are those contained in the legal documents European Union Council Decision 94/904/CE and October 13th Spanish Ministerial Order. The properties studied were the following: flash point, reactivity (gas generation), ecotoxicity, main contaminants extracted by the leaching process and main harmful substances contained in wastes. An additional study of the relationship between flash point and solvent concentration in waste was carried out for polyurethane-acetone and neoprene-toluene systems. The wastes considered were metal containers with remains of dry or semi-dry adhesive. The results obtained show that the presence of solvent in wastes confers on them hazardous characteristics (flash point and harmful composition) depending on the solvent type and its concentration.

Adhesives↗

Airborne endotoxin and its relationship to pulmonary function among workers in an Indian jute mill.

Air samples from various processing areas of an Indian jute mill were examined for endotoxin. The authors assessed work-related respiratory symptoms and pulmonary function changes in the dust-exposed workers from the different processing areas using a standard questionnaire and spirometry. Endotoxin was estimated in water extract of jute dust from 3 milling areas, and in outside air, by the Limulus amebocyte lysate gel clot technique. The batching, spinning, and weaving areas of the jute mill showed endotoxin levels of 0.22-4.42 microg/m3, 0.04-1.47 microg/m3, and 0.01-0.07 microg/m3, respectively, values similar to those found in Indian cotton mills. Respiratory morbidities among the workers included typical byssinotic symptoms, along with acute changes in postshift forced expiratory volume in 1 s (FEV1.0) (31.8%). Results of this study demonstrated that increased exposure to bacterial endotoxin in airborne dust is related to byssinotic symptoms among Indian jute mill workers. Findings were similar to those reported previously for workers in the cotton, flax, and hemp industries.

Air Pollution, Indoor↗

Evaluation of column flotation in the downstream processing of fermentation products: recovery of a genetically engineered alpha-amylase.

Flotation is a simple, inexpensive, and versatile unit operation with a largely unexplored potential in biotechnology. There is a general lack of research concerning biotechnological applications in this area, especially in the recovery of fermentation products. Moreover, the few reports in the literature do not consider the modern concept of column flotation as practiced in the mineral industry. We report herein the application of column flotation for the recovery of a Bacillus stearothermophilus alpha-amylase expressed in Escherichia coli by the use of a food-grade polymer, (hydroxypropyl)methylcellulose (HPMC), and ammonium sulfate. First, the enzyme was removed from the liquid phase by partition to a salted-out HPMC phase. The enzyme-containing polymer flocs were then floated from the liquid. Recovery of active enzyme was as high as 90%, with throughput as high as 94 m3/(h.m2). The floatability of the enzyme from a periplasmic extract was higher than extracellular enzyme in the broth due to the presence of depressors of molecular weight lower than 10,000 in the broth.

Ammonium Sulfate↗

Removal of non-ionic organic pollutants from water via liquid-liquid extraction.

The removal of model pollutants bromocresol green (BG) and phenol from water is demonstrated via two liquid-liquid extraction methods. Both methods exploit selective interactions established by the pollutant molecule with a surfactant, oil, or alcohol, and are variants of the more general Winsor systems where the phases are in contact along an extremely large interfacial area. In the first method the surfactant and the co-surfactant move from a predominantly oil-in-water microemulsion (Winsor I), to a middle phase microemulsion (Winsor III), and finally to a water-in-oil microemulsion (Winsor II), as the physicochemical conditions of salinity, temperature or hydrophilic-lipophilic balance of the surfactant system are varied. This method achieves better than 99% removal of the pollutant BG from water. It is argued that the removal is produced upon increasing the salinity of the system because the interaction of BG with a medium chain-length alcohol drives it to move along with the alcohol to another phase. The second method, which is scalable to industrial levels, uses a spontaneously produced water-in-oil microemulsion with large interfacial area that appears after bringing in contact water and a pre-formed Winsor II or Winsor III microemulsion system containing different surfactants and oils. The method is applied to the removal of phenol from water, and it is found that systems with polar oils such as ethyl butyrate or with cationic surfactants such as stearyl trimethylammonium chloride are more efficient in removing phenol than systems with normal alkanes or anionic surfactants. It is also shown that a microemulsion formed using a polar oil performs better than using only the polar oil as the extraction solvent. Finally, the efficiency of the second liquid-liquid extraction method can be increased from 69% in a single-stage process to 83% in a two-stage process, using the same total amount of extraction solvent.

Alkanes↗

Speciation and recovery of chromium from chromite ore processing residues.

The processing of chromite ore is associated with the generation of large quantities of solid wastes containing chromium, which have been disposed of as landfill for many years. The mobilization and operational speciation of chromium contained in soils contaminated with metal salts are important in terms of the environment. Several methods have been employed for the extraction and recovery of solid wastes. Chromium contained in contaminated soils and solid wastes can be categorized as exchangeable, oxidizable, carbonate-bound, reducible and residual. The results from this study indicate a need for efficient leaching methodologies in chromite ore processing plants to decrease the non-detrital fractions of chromium in the residue. Aggressive methodologies are required to recover chromium from the detrital fractions. The potential benefits of employing sodium peroxide for the complete recovery of chromium from chromite residue have been demonstrated, and the need to ensure the safety of the process has been emphasized.

Chromium↗

Rapid detection of Listeria monocytogenes in ham samples using immunomagnetic separation followed by polymerase chain reaction.

AIMS: To develop a 24-h system for the detection of Listeria monocytogenes in ham. METHODS AND RESULTS: An immunomagnetic separation (IMS) of bacteria directly from ham followed by extraction of DNA and detection using a new multiplex polymerase chain reaction (PCR) was used. The PCR method used one primer pair targeted at the listeriolysin O gene of L. monocytogenes and the other pair for a region of the 23S rRNA genes of Listeria, giving products of 706 and 239 bp, respectively. The combined IMS/PCR was calculated to be capable of detecting as few as 1.1 L. monocytogenes cells g-1 in a 25-g ham sample. CONCLUSION: The process produced acceptable results, but the IMS step is the main barrier to further improvement of sensitivity. The DNA isolation was the most time-consuming step in the process. SIGNIFICANCE AND IMPACT OF THE STUDY: A 24-h test for the presence of L. monocytogenes will be useful to the food industry and significantly assist in the timely investigation of outbreaks.

DNA, Bacterial↗

[Simultaneous isolation of proteins and fats from oil seeds. I. Principle of procedure].

The principle of the procedure for the simultaneous isolation of proteins and fats from oil seeds is illustrated by the isolation of these constituents from sunflower seeds. This procedure is based on the displacement of oil and the extraction of protein by an aqueous electrolyte solution after comminution of the pretreated seeds, removal of the insoluble cell components and separation of the extracts into a fat-containing and a protein-containing phase. The globulins are precipitated isoelectrically; the albumins, by using complexing agents or thermal coagulation. The liberation of the oil from the concentrated fat emulsion is achieved immediately in a mechanical way or after addition of destabilizers. The results are discussed with regard to the yields obtained and the composition of the final products.

Albumins↗