Inhibition of Listeria monocytogenes growth in cured ready-to-eat meat products by use of sodium benzoate and sodium diacetate.

The effect of sodium benzoate (0.08 to 0.25%) in combination with different concentrations of sodium diacetate (0.05 to 0.15%) and NaClI (0.8 to 2%) and different finished product moisture (55 to 75%) on the growth of Listeria monocytogenes in ready-to-eat meat products was evaluated using a central composite design over 18 weeks of storage at 4 degrees C. The effects of these factors on time to growth were analyzed using a time-to-failure regression method. All main effects were significant except product moisture, which was significant when included in the two- and three-way interactions (P < 0.05). Sodium benzoate was more effective (lengthening time to growth) when used with increasing concentrations of sodium diacetate and salt and decreasing finished product moisture. The model indicated that low-moisture products, e.g., bologna or wieners, could have time-to-growth values longer than 18 weeks if they were formulated with 0.1% sodium benzoate and 0.1% sodium diacetate. Time to growth in high-moisture products, e.g., ham or cured turkey breast at 75% moisture, was predicted to be much shorter for the same basic formulation (0.1% sodium benzoate and 0.1% sodium diacetate). Consequently, high-moisture ready-to-eat products in which sodium benzoate is limited to 0.1% (current standard for generally recognized as safe) may need additional ingredients to effectively inhibit growth of L. monocytogenes.

Construction of a fosmid library of cucumber (Cucumis sativus) and comparative analyses of the eIF4E and eIF(iso)4E regions from cucumber and melon (Cucumis melo).

A fosmid library of cucumber was synthesized as an unrestricted resource for researchers and used for comparative sequence analyses to assess synteny between the cucumber and melon genomes, both members of the genus Cucumis and the two most economically important plants in the family Cucurbitaceae. End sequencing of random fosmids produced over 680 kilobases of cucumber genomic sequence, of which 25% was similar to ribosomal DNAs, 25% to satellite sequences, 20% to coding regions in other plants, 4% to transposable elements, 13% to mitochondrial and chloroplast sequences, and 13% showed no hits to the databases. The relatively high frequencies of ribosomal and satellite DNAs are consistent with previous analyses of cucumber DNA. Cucumber fosmids were selected and sequenced that carried eukaryotic initiation factors (eIF) 4E and iso(4E), genes associated with recessively inherited resistances to potyviruses in a number of plants. Indels near eIF4E and eIF(iso)4E mapped independently of the zym, a recessive locus conditioning resistance to Zucchini yellow mosaic virus, establishing that these candidate genes are not zym. Cucumber sequences were compared with melon BACs carrying eIF4E and eIF(iso)4E and revealed extensive sequence conservation and synteny between cucumber and melon across these two independent genomic regions. This high degree of microsynteny will aid in the cloning of orthologous genes from both species, as well as allow for genomic resources developed for one Cucumis species to be used for analyses in other species.

Quantitative trait loci for maysin synthesis in maize (Zea mays L.) lines selected for high silk maysin content.

Maysin is a naturally occurring C-glycosyl flavone found in maize (Zea mays L.) silk tissue that confers resistance to corn earworm (Helicoverpa zea, Boddie). Recently, two new maize populations were derived for high silk maysin. The two populations were named the exotic populations of maize (EPM) and the southern inbreds of maize (SIM). Quantitative trait locus (QTL) analysis was employed to determine which loci were responsible for elevated maysin levels in inbred lines derived from the EPM and SIM populations. The candidate genes consistent with QTL position included the p (pericarp color), c2 (colorless2), whp1 (white pollen1) and in1 (intensifier1) loci. The role of these loci in controlling high maysin levels in silks was tested by expression analysis and use of the loci as genetic markers onto the QTL populations. These studies support p, c2 and whp1, but not in1, as loci controlling maysin. Through this study, we determined that the p locus regulates whp1 transcription and that increased maysin in these inbred lines was primarily due to alleles at both structural and regulatory loci promoting increased flux through the flavone pathway by increasing chalcone synthase activity.

Nerve conduction and sensorineural function in dental hygienists using high frequency ultrasound handpieces.

BACKGROUND: Oscillatory vibration from industrial power tools poses a well-recognized risk of peripheral nerve injury. There have been reports of elevated vibrotactile perception thresholds (VPT) among dentists, dental technicians, and dental hygienists, using rotary devices and ultrasonics. Elevated VPTs are an indicator of small fiber nerve or mechanoreceptor injury, but the high frequencies associated with dental instruments are presumed by the ISO to exceed physiological response upper thresholds. This study examines nerve conduction and sensorineural deficits in dental hygienists. METHODS: A cross-sectional study of 94 experienced dental hygienists was conducted to assess peripheral nerve function and clinical signs and symptoms. Specialized testing included measurement of VPTs for three different categories of mechanoreceptors, sensory nerve conduction tests with fractionated digit and palmar segments, and measurement of calibrated pinch force with force sensitive resistors (FSRs) during a simulated procedure. RESULTS: Chronic hand paresthesias were described by 44.7% of experienced dental hygienists. Sensory nerve conduction velocity (SNCV) across the wrist-palm segment of the median nerve. VPTs were particularly elevated at the FAII mechanoreceptor among experienced dental hygienists. Compared to participants without carpal tunnel syndrome (CTS), as defined by study criteria, 14 experienced hygienists with diagnosed CTS had almost twice the average weekly use of vibratory instruments -8.3 hr versus 4.5 hr, and had SNCV deficits along the digit -47.11 m/sec (+8.70) versus 42.57 m/sec (+8.25), and across the wrist -44.04 m/sec (+7.15) versus 41.36 m/sec (+9.27). There was a distinct subset of dental hygienists (27%) with a combination of low calibrated pinch force in simulations, subjective loss of strength and elevated VPTs, especially in the FAII mechanoreceptor population -110.82 db (+8.57) versus 104.84 db (+6.80) in the rest of the cohort. This subset also had a higher prevalence of paresthesias (67% vs. 39%) and greater cumulative vibration exposure (OR = 1.206 [CI 1.005-1.448]), than other hygienists. CONCLUSIONS: The high levels of paresthesias observed among dental hygienists appear to be attributable to several pathophysiological mechanisms, including, sensory nerve demyelination at the carpal tunnel and intrinsic to the digits, and dysfunction of fingertip mechanoreceptors. A distinct sub-population appears to exhibit a high level of accumulated abnormality.

Occupational contact dermatitis to nickel: experience of the British dermatologists (EPIDERM) and occupational physicians (OPRA) surveillance schemes.

AIMS: To examine, from occupational surveillance reporting data, whether scheme reporters considered nickel exposure to play a role in occupational contact dermatitis (OCD) in the UK. METHODS: Data on occupational skin disease in the UK are collected by two occupational disease surveillance schemes, EPIDERM and OPRA. Cases of OCD believed to have relevant nickel exposure reported to EPIDERM or OPRA from February 1993 to January 1999 were studied. RESULTS: An estimate of 1190 cases of occupational contact dermatitis thought to have relevant nickel exposure (12% of total estimated OCD) was derived from reports by dermatologists, an average of 198 per year. The highest incidence rates were seen in hairdressers (23.9/100 000 workers/year), bar staff (4.7), chefs and cooks (4.4), retail cash and checkout operators (2.8), and catering assistants (2.5). From May 1994 to January 1999, 158 cases of nickel associated dermatitis (1.9% of total OCD cases) were estimated; the most frequently reported occupations were electronic assemblers, nurses, sales assistants, and general assemblers. From July 1997 to January 1999, 547 positive patch tests to nickel were reported; in 195 cases (36%), nickel was felt to be a relevant occupational allergen (for example, coin handling). In hairdressers, nurses, cooks, and beauticians, nickel was usually considered, if relevant at all, to be only one of several causes of dermatitis. CONCLUSIONS: Up to 12% of total estimated cases of OCD were thought to be due in part to nickel. Results suggest that nickel hypersensitivity is one of several contributors to OCD in subjects with multiple occupational exposures. Coin handling may be a source of OCD to nickel.

Inhibition of nonproteolytic, psychrotrophic clostridia and anaerobic sporeformers by sodium diacetate and sodium lactate in cook-in-bag turkey breast.

A nonproteolytic, psychrotrophic Clostridium isolate, designated strain OMFRI1, was recovered from cook-in-bag turkey breasts (CIBTB) that displayed an intense pink discoloration and an off-odor following extended refrigerated storage. The viability of strain OMFRI1 in CIBTB containing sodium diacetate (at 0, 0.25, and 0.5%) and/or sodium lactate (at 0, 1.25, and 2.5%) was subsequently evaluated. Raw CIBTB batter was inoculated with 9 to 30 spores of strain OMFRI1 per g, vacuum packaged, cooked to an instantaneous internal temperature of 71.1 degrees C, chilled, and incubated at 4 degrees C for up to 22 weeks. In the absence of food-grade antimicrobial agents, spoilage (i.e., an off-odor) occurred within 6 weeks, and anaerobic plate counts reached 6.6 log10 CFU/g. The CIBTB containing sodium diacetate (0.25%) and that containing sodium lactate (1.25%) required 12 weeks for spoilage to occur and for anaerobic plate counts to reach 7.0 and 6.0 log10 CFU/g, respectively. When sodium diacetate (0.25%) and sodium lactate (1.25%) were used in combination, no off-odor was detected and anaerobic plate counts did not exceed 2.3 log10 CFU/g over 22 weeks of storage at 4 degrees C. In related experiments, sodium diacetate (at 0, 0.25, and 0.5%), sodium lactate (at 0, 1.25, and 2.5%), and combinations of both ingredients were evaluated in uninoculated CIBTB incubated at 25 degrees C for up to 22 days. In the absence of antimicrobial agents and in CIBTB containing sodium diacetate (0.5%), spoilage occurred within 8 days and anaerobic plate counts reached 6.8 and 6.6 log10 CFU/g, respectively. Samples of CIBTB containing sodium lactate (2.5%) showed signs of spoilage within 22 days, and anaerobic plate counts for these samples ranged from < or = 1.0 to 6.3 log10 CFU/g. In CIBTB containing both sodium lactate (2.5%) and sodium diacetate (0.25%), spoilage was not evident and anaerobic plate counts were < or = 1.0 log10 CFU/g within 22 days. These data validate the efficacy of sodium lactate and sodium diacetate in extending the shelf life of CIBTB.

Characterization of the solution conformations of leuprolide acetate.

Leuprolide acetate (pGlu-His-Trp-Ser-Tyr-d-Leu-Leu-Arg-Pro-NHEt), a potent LHRH agonist in wide clinical use, was characterized conformationally by NMR and circular dichroism. It displayed quite different preferred conformations under different solution conditions: two low population beta-turns in water, a nascent helix in TFE/water at low pH, and a high population beta-turn in TFE/water at slightly acidic pH. The pH-related conformational change in TFE/water is attributed to the pK(a) of the acetate counterion, not to ionizable groups on the peptide. None of these conformations are in exact agreement with previous computational predictions.

Prognosis and work absence due to occupational contact dermatitis.

The consequences of a diagnosis of occupational contact dermatitis (OCD) were investigated using cases available from the voluntary surveillance scheme, EPIDERM. Cases of OCD reported from November 1994 to September 1995 were identified and sampled to give at least 100 cases of allergic, irritant and mixed OCD reported by consultant dermatologists (344 cases) and occupational physicians (377 cases). A questionnaire was sent to the reporting physician to elicit further information. 512 completed questionnaires were returned, of which 510 were eligible for analysis. Among cases reported by dermatologists (n = 286) and occupational physicians (n = 224), 7% (6.3%) had been unemployed and 16.8% (20.1%) had taken sick leave. 3 factors independently predicted time off work in a logistic regression analysis: age OR = 1.25 (95% CI, 1.05-1.49), allergic dermatitis OR = 1.77 (95% CI, 1.13-2.79) and medicolegal assessment OR = 4.42 (95% CI, 2.20-8.89). Overall, 15.7% did not improve clinically between the first and last visit. Those who did not improve had been exposed to the agent for longer (mean 7.6 years) than those who did (5.3 years) (p = 0.09). In patients

Surveillance for work-related hearing loss in the UK: OSSA and OPRA 1997-2000.

Surveillance data on occupational audiological disorders have been collected by the Occupational Surveillance Scheme for Audiological Physicians (OSSA) since October 1997 and by the Occupational Physicians Reporting Activity (OPRA) from January 1996. During the 3 years ending in September 2000, a total of 1620 new cases were received from consultant audiological physicians; 988 new cases were estimated from reports by occupational physicians in the period from October 1997 to September 2000. The annual incidence of occupational noise-induced hearing loss (NIHL) was 1.94 and 1.23 per 100 000 workers for the OSSA and OPRA schemes, respectively. The median age at diagnosis with NIHL was 59 years in OSSA reports and 50 years in OPRA; nearly all cases were seen in men (95.6 and 92.5% male cases for OSSA and OPRA, respectively). High incidence rates based on OSSA reports were seen in foundry labourers (64.0 per 100 000 employed), coal gas and coke oven furnace workers (54.6), workers in transport and communication (43.1), metal workers (31.3), and members of the armed forces (28.3). Data from occupational physicians point to high rates in sawyers and woodworking machinists, metal furnace workers, coach and carriage builders, maintenance fitters, and engineering labourers. Among workers aged < or =45 years, those in manufacturing and the armed forces were prominent. The long latency of occupational hearing loss makes surveillance difficult, but consistent patterns in occupational risk suggest targets for preventive efforts.

[Iliopsoas impingement after cementless total hip arthroplasty. Case reports]. / Iliopsoaskontaktphänomen nach zementfreier Hüfttotalendoprothese. Eine Fallbeschreibung.

Postoperative pain after total hip arthroplasty can be caused by several conditions. We describe two cases of postoperative persistent iliopsoas impingement after cementless implantation of the acetabular component. The groin pain was relieved after revision with a more anteverted acetabular component. We describe the diagnostic methods as well as the principles of the revision.

Modeling the growth of Listeria monocytogenes in cured ready-to-eat processed meat products by manipulation of sodium chloride, sodium diacetate, potassium lactate, and product moisture content.

A central composite second-order response surface design was employed to determine the influences of added sodium chloride (0.8 to 3.6%), sodium diacetate (0 to 0.2%), potassium lactate syrup (0.25 to 9.25%), and finished-product moisture (45.5 to 83.5%) on the predicted growth rate of Listeria monocytogenes in cured ready-to-eat (RTE) meat products. Increased amounts of both sodium diacetate (P < 0.11) and potassium lactate (P < 0.001) resulted in significant reductions in the growth rate constants of L monocytogenes. Increased finished-product moisture (P < 0.11) significantly increased growth rate constants. The nfluence of sodium chloride was not statistically significant. The second-order statistical factor for lactate was significant (P < 0.01), but all two-way interactions were not. In general, predicted growth rates exceeded actual growth rates obtained from inoculation studies of four cured RTE meat products (wieners, smoked-cooked ham, light bologna, and cotto salami). The final model will be useful to food technologists in determining formulations that will result in finished cured RTE meat products in which L. monocytogenes is not likely to grow.

The reported incidence of work-related musculoskeletal disease in the UK: MOSS 1997-2000.

Consultant rheumatologists participate in surveillance of work-related musculoskeletal conditions under the Musculoskeletal Occupational Surveillance Scheme (MOSS), which has been in operation since 1997. During the first 3 years of the scheme, an estimated total of 8070 cases and 8442 diagnoses were obtained, an average of slightly less than 2700 estimated cases each year. Disorders of the upper limb accounted for approximately 66% (5502) of the total, with hand/wrist/arm conditions (3693 cases) comprising the majority of these. Conditions of the lumbar spine and trunk (13% of cases), the cervical spine (12%) and the shoulder (12%) were also frequently reported. Pain with ill-defined pathology was reported in 35% of cases with hand and forearm disorders. Overall, 82% of cases were related to repetitive rather than single injury. The largest numbers of cases were seen in workers in craft occupations (1659) and in clerical and secretarial workers (1524). High rates of musculoskeletal conditions, particularly of upper limb disorders, are notable in mining. In most occupations, and overall, women were at greater risk than men.

Spark-source mass spectrometric assessment of silicon concentrations in silicon-doped gallium arsenide single crystals.

The spark-source mass spectrometric assessment of silicon concentrations in silicon-doped vertical-gradient-freeze gallium arsenide is presented. The silicon concentrations determined are compared with the charge-carrier densities measured by means of the Hall effect with van der Pauw symmetry along the axis of a single crystal.

SWORD '99: surveillance of work-related and occupational respiratory disease in the UK.

Systematic reports from chest and occupational physicians under the SWORD and OPRA (Occupational Physicians Reporting Activity) surveillance schemes continue to provide a picture of the incidence of occupational respiratory disease in the UK. An estimated total of 4393 incident cases (comprising 4530 diagnoses) were reported during the 1999 calendar year, an increase of 1427 cases over the previous year. Benign pleural disease was the single most frequently reported condition (28% of all diagnoses reported). Occupational asthma cases (1168; 26%) remained high, as did mesothelioma (1032; 23%). Analysis of trends over the past 8 years shows an increase in mesothelioma cases, but little change in asthma. The annual incidence per 100,000 employed people, 1996-1999, for mesothelioma, lung cancer and pneumoconiosis was high amongst construction workers (28.7), miners and quarrymen (26.5), woodworkers (18.9) and gas, coal and chemical workers (15.2). Trends in mesothelioma incidence by birth cohort continue to show an increase in construction workers and a continuing decline in shipyard and insulation workers. The relative proportion of pneumoconiosis cases attributed to coal mining has fallen steadily in workers born since approximately 1920 and most cases are now in men who have been employed in quarrying and rock drilling.

Comparative fourier transform infrared and circular dichroism spectroscopic analysis of alpha1-proteinase inhibitor and ovalbumin in aqueous solution.

Alpha1-proteinase inhibitor (alpha1Pi) and ovalbumin are both members of the serpin superfamily. They share about a 30% sequence identity and exhibit great similarity in their three-dimensional structures. However, no apparent functional relationship has been found between the two proteins. Unlike alpha1Pi, ovalbumin shows no inhibitory effect to serine proteases. To see whether or not a conformational factor(s) may contribute to the functional difference, we carried out comparative analysis of the two proteins' secondary structure, thermal stability, and H-D exchange using FT-IR and CD spectroscopy. FT-IR analysis reveals significant differences in the amide I spectral patterns of the two proteins. Upon thermal denaturation, both proteins exhibit a strong low-wavenumber beta-sheet band at 1624 cm(-1) and a weak high-wavenumber beta-sheet band at 1694 cm(-1), indicative of intermolecular aggregate formation. However, the midpoint of the thermal-induced transition of alpha1Pi (approximately 55 degrees C) is 18 degrees C lower than that of ovalbumin (approximately 73 degrees C). The thermal stability analysis provides new insight into the structural changes associated with denaturation. The result of H-D exchange explains some puzzling spectral differences between the two proteins in D2O reported previously.

Effect of zinc binding and precipitation on structures of recombinant human growth hormone and nerve growth factor.

Metal-induced precipitation of protein therapeutics is being used and further developed as a processing step in protein formulation and may have utility in protein purification and bulk storage. In such processes, it is imperative that native protein structure is maintained and the metal complexation is reversible. In the current study, we investigated the effects of zinc-induced precipitation on recombinant human growth hormone (rhGH) and recombinant human nerve growth factor (rhNGF). On the addition of ethylenediaminetetraacetic acid (EDTA), the precipitates were dissolved, yielding complete recovery of native protein in both cases. Both proteins have specific metal binding sites and require specific molar ratios of zinc to protein to initiate precipitation (zinc:rhGH > 2:1; zinc:rhNGF > 18:1). Furthermore, the secondary structures of both proteins were unperturbed in soluble zinc complexes and zinc-induced precipitates, as measured by infrared and circular dichroism spectroscopies. The soluble zinc complex of rhGH had minor tertiary structural alterations, whereas zinc binding did not alter the tertiary structure of rhNGF. These studies indicated that metal-induced precipitation provides a method to maintain proteins in their native state in precipitates, which may be useful for purification, storage, and formulation.

Occupational contact dermatitis in the UK: a surveillance report from EPIDERM and OPRA.

Since February 1993 the EPIDERM surveillance scheme has collected data on occupational skin disease from consultant dermatologists in the UK. Reporting by occupational physicians to the scheme began in May 1994 and was superseded in January 1996 by the Occupational Physicians Reporting Activity (OPRA). The schemes currently receive reports on incident cases from 244 dermatologists and 790 occupational physicians. An estimated total of 9937 cases of contact dermatitis reported by dermatologists was calculated from surveillance data; 8129 contact dermatitis cases were estimated from reports by occupational physicians. The annual incidence of occupational contact dermatitis from dermatologist reports was 6.4 cases per 100,000 workers and 6.5 per 100,000 from reports by occupational physicians, an overall rate of 12.9 cases per 100,000 workers. Manufacturing industries account for the greatest number of cases seen by both sets of reporting physicians, with health care employment second. Reports from dermatologists also indicate high rates of dermatitis in the personal service industries (mainly hairdressers and barbers) and in agriculture. With the exception of an increase in cases seen in nurses in both schemes, the numbers and proportions of cases of contact dermatitis within occupations have remained fairly constant over the 6-year reporting period. Agents accounting for the highest number of allergic contact dermatitis cases were rubber (23.4% of allergic cases reported by dermatologists), nickel (18.2), epoxies and other resins (15.6), aromatic amines (8.6), chromium and chromates (8.1), fragrances and cosmetics (8.0), and preservatives (7.3). Soaps (22.0% of cases), wet work (19.8), petroleum products (8.7), solvents (8.0), and cutting oils and coolants (7.8) were the most frequently cited agents in cases of irritant dermatitis. The national scope of the data, together with the parallel structure by which both dermatologists and occupational physicians report incident cases, is useful in determining the extent of skin hazards in UK industry and may help in better targeting efforts to reduce the burden of skin disease at work.

Surveillance of occupational skin disease: EPIDERM and OPRA.

Consultant dermatologists in the U.K. have been reporting to EPIDERM, a voluntary surveillance scheme for occupational skin disease, since February 1993; reporting by occupational physicians to the scheme began in May 1994 and was superseded in January 1996 by OPRA (Occupational Physicians Reporting Activity). Currently 244 dermatologists and 790 occupational physicians report incident cases to these schemes. During the 6 years to January 1999 a total of 12, 574 new cases of occupational skin disease was estimated from reports by consultant dermatologists and 10,136 cases estimated from occupational physicians (since May 1994). The annual incidence of occupational contact dermatitis using data from both schemes was 12. 9 per 100,000 workers. The incidence of contact dermatitis per 100, 000 workers increased with age in men from 4.9 (age 16-29 years) to 6.6 (age 45-60 years); in women a higher rate (9.5) was apparent in the younger age group, with lower rates in older female workers. High rates in young workers were associated with wet work and in older workers with exposure to oils. For men, high rates of contact dermatitis were seen in reports from both schemes for chemical operatives, machine tool setters and operatives, coach and spray painters and metal workers. For women, high rates were found for hairdressers, biological scientists and laboratory workers, nurses and those working in catering. The most frequent agents for contact dermatitis were rubber chemicals and materials (14.1% of cases reported by dermatologists), soaps and cleaners (12.7%), nickel (11. 9%), wet work (11.1%), personal protective equipment (6.2%), petroleum products (6.3%), cutting oils and coolants (5.6%), and epoxy and other resins (6.1%). In the 1608 estimated cases of skin cancer all but 4% were attributed to ultraviolet radiation. Cases of contact urticaria attributed to latex peaked in 1996, with a decline in cases since that time.

Stability of human serum albumin during bioprocessing: denaturation and aggregation during processing of albumin paste.

PURPOSE: To assess the impact of various bioprocessing steps on the stability of freshly precipitated human serum albumin (HSA) obtained from pooled human plasma. METHODS: After initial precipitation of HSA from plasma, the resultant paste is either (a) lyophilized or (b) washed with acetone and then air-dried in order to obtain a dry powder. The structure of HSA was examined using Fourier transform infrared (IR) spectroscopy. The extent of aggregation of redissolved HSA was measured using both dynamic light scattering and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: Both lyophilization and air-drying perturb the secondary structural composition of HSA, as detected by infrared (IR) spectroscopy. Upon dissolution of dried paste, most of the protein refolds to a native-like conformation. However, a small fraction of the protein molecules form soluble aggregates that can be detected by both dynamic light scattering and SDS-PAGE. The level of aggregation is so low that it could not be detected in the bulk by either circular dichroism or IR spectroscopy. The lyophilized protein, which appears to be more unfolded in the solid state than the acetone washed/air-dried material, exhibits a higher level of aggregation upon dissolution. CONCLUSIONS: There is a direct correlation between the extent of unfolding in the solid state and the amount of soluble aggregate present after dissolution. Moreover, the presence of the aggregates persists throughout the remainder of the purification process, which includes dissolution, chromatography, sterile filtration and viral inactivation steps. Analytical methods used to monitor the stability of biopharmaceuticals in the final product can be used to assess damage inflicted during processing of protein pharmaceuticals.

Long chain arginine esters: a new class of cationic detergents for preparation of hydrophobic ion-paired complexes.

The ability of stoichiometric amounts (based on charged groups) of ionic detergents to bind to oppositely charged ionic compounds has been recently reviewed. These hydrophobic ion-paired (HIP) complexes display altered solubility properties. Most of the work to date on HIP compelxes has focused on basic drugs and anionic detergents. It would be extremely useful to extend this approach to acidic compounds, including DNA and RNA. However, most cationic detergents are relatively toxic. It is hypothesized that detergents constructed from naturally occurring or well tolerated components, coupled by labile linkages, will be less toxic and still able to form strong HIP complexes. This study describes the synthesis and characterization of long chain alkyl esters of arginine. This class of cationic detergents, which have not been reported previously, are less cytotoxic than alkyltrimethylammonium detergents, possibly making them more acceptable in drug delivery applications. These arginine esters exhibit detergent-like properties. For example, the dodecyl ester of arginine has a critical micelle concentration of 0.07 mM, while being approximately 5-10 fold less toxic than tetradecyltrimethylammonium bromide. The arginine dodecyl ester forms stable HIP complexes with plasmid DNA. The complex is sufficiently stable to allow some modest level of transfection with Cos-7 cells in a time- and concentration-dependent fashion. This work demonstrates that arginine-based cationic detergents are effective ion-pairing agents, appear to be less toxic than alkyltrimethylammonium compounds, and form stable complexes with DNA.