Polycyclic aromatic hydrocarbons in human milk of nonsmoking U.S. women

The determination of polycyclic aromatic hydrocarbons (PAHs) in human milk is relevant in assessing the dose to mothers and infants. Since measurements for the United States are lacking, we analyzed 13 PAHs in human milk from 12 nonsmoking American mothers. Analytes were extracted from milk by using a combination of centrifugation, liquid-liquid, and solid-phase extraction techniques. Following method validation with spiked bovine milk, PAH concentrations in human milk were determined by isotope dilution gas chromatography/mass spectrometry. Phenanthrene, fluorene, fluoranthene, and pyrene were found at median concentrations of 11.8, 3.0, 1.6, and 1.1 ng/g milk fat, respectively, whereas 9 additional analytes including benzo[a]pyrene were not detected (<0.04 ng/g). Calculated average daily doses from ingestion (high: 75.8, phenanthrene; low: 6.1 ng/kg/day pyrene) were compared to doses from air pollution inhalation. Milk ingestion accounted for 47.9-81.4% of the total dose of individual PAHs to the infant. However, calculated total average daily doses of phenanthrene and pyrene from ingestion and inhalation combined (12.2 to 110.3 ng/kg-day, respectively) were 3 orders of magnitude lower than documented doses known to cause health effects. These first measurements of PAH levels in U.S. human milk are comparable to concentrations reported previously for European and Asian women.     

Evaluation of standard and new chromogenic selective plating media for isolation and identification of Bacillus cereus

In this study, the performance of two new chromogenic plating media (CBC and BCM) was compared with two standard selective plating media (PEMBA and MYP) recommended by food authorities for isolation, identification and enumeration of Bacillus cereus. The four media types were challenged with a strain set comprising 100 B. cereus isolates from different origins and with different toxigenic potentials (40 food isolates, 40 isolates from food borne outbreaks and 20 clinical isolates). Additionally, the performance of the plating media for analysis of complex samples was assessed using naturally contaminated foods. Our survey showed that the new chromogenic media represent a good alternative to the conventional standard media. Especially, if laboratory staff are not highly trained in identification of B. cereus, the conventional media could lead to substantial misidentification and underestimation of food borne illness caused by B. cereus. However, there are some B. cereus strains that could not even be detected with this new type of chromogenic media. After the fatal misidentification of a highly toxic strain, other methods for a conclusive identification of B. cereus are needed. Sequence analysis of the plcR gene, a pleiotropic regulator of various virulence factors and B. cereus specific enzymes, revealed a significant correlation between atypical colony appearance and specific variances within the plcR gene sequences of those strains. The current concept of selective plating media, utilising PlcR regulated enzyme activities for differentiation purposes, should therefore be reconsidered and research should be geared towards culture independent methods.     

Use of nisin-coated plastic films to control Listeria monocytogenes on vacuum-packaged cold-smoked salmon

Cold-smoked (Salmo salar) salmon samples were surface-inoculated with a cocktail of three nisin-resistant strains of L. monocytogenes (PSU1, PSU2 and PSU21) to a level of approximately 5 x 10(2) or 5 x 10(5) CFU/cm2 of salmon surface. The inoculated smoked salmon samples were vacuum-packaged with control film (no nisin) or nisin-coated plastic films and stored at either 4 or 10 degrees C. When the inoculated smoked salmon samples were packaged with film coated with 2000 IU/cm2 of nisin, a reduction of 3.9 log CFU/cm2 (compared with control) was achieved at either temperature for samples inoculated with 5 x 10(2) CFU/cm(2 of L. monocytogenes after 56 (4 degrees C) and 49 (10 degrees C) days of storage while reductions of 2.4 and 0.7 log CFU/cm2 were achieved for samples inoculated with a high level of L. monocytogenes (5 x 10(5) CFU/cm2) after 58 (4 degrees C) and 43 (10 degrees C) days, respectively. For samples packaged in film coated with 500 IU/cm2 of nisin, reductions of 0.5 and 1.7 log CFU/cm2 were achieved for samples inoculated with a low level of L. monocytogenes (5 x 10(2) CFU/cm2) after 56 (4 degrees C) and 49 (10 degrees C) days of storage while reductions of 1.8 and 0.8 log CFU/cm2 were achieved for samples inoculated with high level of L. monocytogenes after 58(4 degrees C) and 43 (10 degrees C) days, respectively. In addition, nisin inhibited the proliferation of background microbiota on smoked salmon in a concentration-dependent manner at both storage temperatures although the bacteriostatic effect was more pronounced at refrigeration temperature. This work highlights the potential for incorporating nisin into plastic films for enhancing the microbial safety of smoked salmon as well as controlling its microbial spoilage.     

Survey of retail alfalfa sprouts and mushrooms for the presence of Escherichia coil O157:H7, Salmonella,and Listeria with BAX,and evaluation of this polymerase chain reaction-based system with experimentally contaminated samples

BAX, a polymerase chain reaction (PCR)-based pathogen detection system, was used to survey retail sprouts and mushrooms for contamination with Escherichia coli O157:H7, Salmonella, Listeria spp., and Listeria monocytogenes. No Salmonella or E. coli O157:H7 was detected in the 202 mushroom and 206 alfalfa sprout samples screened. L. monocytogenes was detected in one sprout sample, and seven additional sprout samples tested positive for the genus Listeria. BAX also detected Listeria species in 17 of the mushroom samples. Only 6 of 850 PCR assays (0.7%) failed to amplify control DNA, and therefore reagent failures and the inhibition of PCR by plant compounds were rare. The sensitivity of the detection system was evaluated by assaying samples inoculated with 10 CFU of each of the pathogens. One hundred seventy-two alfalfa sprout samples were inoculated with E. coli O157:H7, and two sets of 130 samples were experimentally contaminated with Salmonella Enteritidis and L. monocytogenes. The frequency of detection depended on the protocols used for inoculation and culturing. Inoculation of samples with approximately 10 CFU from frozen stocks yielded detection rates of 87.5 and 94.5% for L. monocylogenes and Salmonella Enteritidis, respectively, in mushrooms. The corresponding rates for alfalfa sprouts were 94.5 and 76.3%. The E. coli O157:H7 detection rate was 100% for mushrooms but only 48.6% for sprouts when standard BAX culture protocols were used. The substitution of an overnight incubation in modified E. coli medium for the 3-h brain heart infusion incubation increased the rate of E. coli O157:H7 detection to 75% for experimentally contaminated sprouts. The detection rate was 100% when E. coli O157:H7 cells from a fresh overnight culture were used for the inoculation. Test sensitivity is therefore influenced by the type of produce involved and is probably related to the growth of pathogens in the resuscitation and enrichment media.     

An interactive data management system for river water quality data

Water quality data banks are needed to document the status and trends of water pollution in a country. Examples of such systems are the STORET system in the U.S., the NAQUADAT system in Canada and the EIS system in Scandinavia. All of these systems require trained personnel to help in the formulation of the inquiry and the actual querying of the system.By contrast, what is described in this paper is an on-line, interactive data management and analysis system which allows the user the direct search, update, retrieval and analysis of the data from a computer terminal. The user addresses the system in a high level language closely resembling English and has complete control over building, updating and querying the individual data banks. Almost all statistical operations can be performed on the data starting from histograms, distributions, correlations to regression, discriminant, component and spectral analysis. Commands for producing camera-ready graphs on graphic terminals are available.The system is implemented on The University of Michigan Computer System and can be accessed through local telephone numbers in more than 100 cities in the U.S. and Canada and from the major European capitals via the TELENET system. The operation of the system is illustrated on a small sample data base on the Ohio river provided by the Ohio River Sanitation Commission (ORSANCO).     

Development and application of DNA analytical methods for the detection of GMOs in food

The principle of direct detection of recombinant DNA in food by the polymerase chain reaction (PCR) is discussed following the three main steps: DNA-extraction, amplification by PCR and verification of PCR products.

Suitable methods for genomic DNA isolation from homogenous, heterogeneous, low DNA containing matrices (e.g. lecithin), gelatinising material (e.g. starch), derivatives and finished products based on classical protocols and/or a combination with commercially available extraction kits are discussed. Various factors contribute to the degradation of DNA such as hydrolysis due to prolonged heat-treatment, nuclease activity and increased depurination and hydrolysis at low pH. The term “DNA quality” is defined as the degree of degradation of DNA (fragment size less than 400 bp in highly processed food) and by the presence or absence of potent inhibitors of the PCR and is, therefore, a key criterion. In general, no DNA is detectable in highly heat-treated food products, hydrolysed plant proteins (e.g. soya sauce), purified lecithin, starch derivatives (e.g. maltodextrins, glucose syrup) and defined chemical substances such as refined soya oil.

If the nucleotide sequence of a target gene or stretch of transgenic DNA is already known specific primers can be synthesised and the segment of rDNA amplified. Detection limits are in the range 20 pg–10 ng target DNA and 0.0001–1% mass fraction of GMO. Amplification products are then separated by agarose gel electrophoresis and the expected fragment size estimated by comparison with a DNA molecular weight marker.

Several methods are used to verify PCR results and they vary in reliability, precision and cost. They include specific cleavage of the amplification products by restriction endonucleases or the more time-consuming, but also more specific, transfer of separated PCR-products onto membranes (Southern Blot) followed by hybridisation with a DNA probe specific for the target sequence. Alternatively, PCR products may be verified by direct sequencing. Nested-PCR assays combines high specificity and sensitivity.

Methods for the screening of 35S-promoter, NOS-terminator and other marker genes used in a wide range of GMOs, the specific detection of approved products such as FlavrSavr™ tomatoes, Roundup Ready™ Soya, Bt-maize 176 and official validated methods for potatoes and genetically modified micro-organisms, that have a model character, are available. Methods to analyse new GMO products are being validated by interlaboratory tests and new techniques are in development (e.g. EC project: DMIF-GEN). However, these efforts may be hampered by the lack of availability of GMO reference material as well as specific sequence information which so far can only be obtained from the suppliers.     

Disclosure: Chaotic implementation thwarts intention of directive

Jan Willem Langeraar of Ecofys and Rolf de Vos of Greenprices argue that, as well as the Guarantee of Origin (see Refocus, Jul/Aug 2003, p. 62), European harmonisation is also needed for the mandatory disclosure of the fuel mix (legislation to be implemented before July 1st 2004). This they claim is needed foremost in the monitoring of the environmental quality of power generation. A harmonised way of collecting information will guarantee transparency and - in the end - establish the consumer's trust. There are several initiatives currently on the table. The implementation of the disclosure in Austria shows what the harmonisation could look like.     

A hardware accelerated implementation of the IEEE 802.15.3 MAC protocol

We present a hardware/software implementation of the IEEE 802.15.3 MAC protocol. Processing-intensive and time-critical protocol tasks are handled by a protocol accelerator that is integrated on-chip with a 32-bit general-purpose processor in order to achieve a moderate (20–40 MHz) system clock frequency. This enables low-power wireless devices compliant with this standard, providing high data rate, multimedia communication. One of the main tasks of the protocol accelerator is to analyze received or transmitted beacons. Based on the channel time allocations broadcast in the beacon and frame information stored in a hardware transmission queue, frames are transmitted without immediate control of the processor. Other features of the protocol accelerator include CRC generation, handling of immediate acknowledgment frames, and direct memory access.     

Affect-focused body psychotherapy in patients with generalized anxiety disorder: Evaluation of an integrative method.

The aim of this study was to explore the long-term effects of affect-focused body psychotherapy (ABP) for patients with generalized anxiety disorder (GAD). A group of 61 consecutive patients, 21–55 years old, were randomized to ABP and psychiatric treatment as usual (TAU). The patients were assessed before treatment and followed up 1 and 2 years after inclusion. The ABP patients received one session of treatment per week during 1 year. Three self-report questionnaires were administered; Symptom Checklist—90, Beck Anxiety Inventory, and the WHO (Ten) Well-Being Index. In both groups, there was a significant improvement. On termination, the ABP group had improved significantly more on the SCL-90 Global Symptom Index than the TAU group, whereas the differences were short of significance on the other two scales. The integration of bodily techniques with a focus on affects in a psychodynamically informed treatment seems to be a viable treatment alternative for patients with GAD. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Influence of sputter deposition parameters on the properties of tunable barium strontium titanate thin films for microwave applications

Barium strontium titanate (BST) thin films are studied with respect to their application as tunable dielectric at microwave frequencies. BST thin films are deposited by means of radio-frequency magnetron sputtering on platinized Si substrates. The substrate to target distance during sputter deposition is varied and the effect on structure, topology, composition and electronic properties is monitored using X-ray diffraction, atomic force microscopy, Rutherford backscattering spectrometry and X-ray photoelectron spectroscopy. These findings are related to the dielectric measurements, which are carried out at 1 MHz and in the microwave range up to 8 GHz using metal-insulator-metal structures with Pt electrodes. For further device evaluation, leakage current measurements are carried out. Changing the process parameter strongly affects the composition of the films. The results emphasize the possibility for enhancing the microwave properties by fine-tuning of the chosen process parameter.