Stable production of hyaluronic acid in Streptococcus zooppidemicus chemostats operated at high dilution rate

Hyaluronic acid is routinely produced through fermentation of both Group A and C streptococci. Despite significant production costs associated with short fermentations and removal of contaminating proteins released during entry into stationary phase, hyaluronic acid is typically produced in batch rather than continuous culture. The main reason is that hyaluronic acid synthesis has been found to be unstable in continuous culture except at very low dilution rates. Here, we investigated the mechanisms underlying this instability and developed a stable, high dilution rate (0.4 h(-1)) chemostat process for both chemically defined and complex media operating for more than 150 h of production. In chemically defined medium, the product yield was 25% higher in chemostat cultures than in conventional batch culture when arginine or glucose was the limiting substrate. In contrast, glutamine limitation resulted in higher ATP requirements and a yield similar to that observed in batch culture. In complex, glucose-limited medium, ATP requirements were greatly reduced but biomass synthesis was favored over hyaluronic acid and no improvement in hyaluronic acid yield was observed. The successful establishment of continuous culture at high dilution rate enables both commercial production at reduced cost and a more rational characterization and optimization of hyaluronic acid production in streptococci. (c) 2005 Wiley Periodicals, Inc.

Muscle coordination during rapid force production by young and older adults

Background. Older adults typically exhibit dramatic reductions in the rate of force development and deficits in the execution of rapid coordinated movements. The purpose of the current study was to investigate the association between the reduced rate of force development exhibited by older adults and the ability to coordinate groups of muscles. Methods. The performance of a visually guided aiming task that required the generation of isometric torque about the elbow joint was compared in 10 young adults (age range, 19 to 29 years) and 10 older adults (age range, 65 to 80 years). Participants were required to exert isometric torque in flexion, extension, pronation, or supination, or in combinations of these directions, to reach a target in minimum time. Surface electromyograms were obtained from the biceps brachii, triceps brachii, brachioradialis, and flexor carpi radialis. Results. Older participants exhibited slower target acquisition times compared with young participants (p < .05), with the extent of the differences between the groups varying markedly between target locations. Conclusions. The impairment in performance, although partially attributable to a general decline in the ability to produce force rapidly, was also affected by the requirements for muscular coordination. At the neuromuscular level, differences between the young and the elderly were expressed most prominently in the bifunctional muscle biceps brachii and in certain temporal aspects of muscular coordination.

Plant-based immunocontraceptive control of wildlife - "potentials, limitations, and possums"

Possums (Trichosurus vulpecula), originally introduced from Australia, are spread over 90% of New Zealand and cause major economic and environmental damage. Immunocontraception has been suggested as a humane means to control them. Marsupial-specific reproductive antigens expressed at high levels in edible transgenic plant tissue might provide a safe, effective, and cheap oral delivery bait for immuno-contraceptive control. As proof of concept, female possums vaccinated with immunocontraceptive antigens showed reduced fertility, and possums fed with potato-expressed heat labile toxin-B (LT-B) had mucosal and systemic immune responses to the antigen. This demonstrated that immunocontraception was effective in possums and that oral delivery in edible plant material might be possible. Nuclear transformation with reporter genes showed that transgenic carrot roots accumulate high levels of foreign protein in edible tissues, indicating their potential as a delivery vector. However, prior to attempts at large scale production, more effective immunocontraceptive antigen-adjuvant formulations are probably required before plant-based immunocontraception can become a major tool for immunocontraceptive control of overabundant vertebrate pests. (c) 2004 Elsevier Ltd. All rights reserved.

Microbial hyaluronic acid production

Hyaluronic acid (HA) is a commercially valuable medical biopolymer increasingly produced through microbial fermentation. Viscosity limits product yield and the focus of research and development has been on improving the key quality parameters, purity and molecular weight. Traditional strain and process optimisation has yielded significant improvements, but appears to have reached a limit. Metabolic engineering is providing new opportunities and HA produced in a heterologous host is about to enter the market. In order to realise the full potential of metabolic engineering, however, greater understanding of the mechanisms underlying chain termination is required.

Co-opting conservation: Migrant resource control and access to national park management in the Philippine uplands

The history of political and economic inequality in forest villages can shape how and why resource use conflicts arise during the evolution of national parks management. In the Philippine uplands, indigenous peoples and migrant settlers co-exist, compete over land and forest resources, and shape how managers preserve forests through national parks. This article examines how migrants have claimed lands and changed production and exchange relations among the indigenous Tagbanua to build on and benefit from otherwise coercive park management on Palawan Island, the Philippines. Migrant control over productive resources has influenced who, within each group, could sustain agriculture in the face of the state's dominant conservation narrative - valorizing migrant paddy rice and criminalizing Tagbanua swiddens. Upon settling, migrant farmers used new political and economic strengths to tap into provincial political networks in order to be hired at a national park. As a result, they were able to steer management to support paddy rice at the expense of swidden cultivation. While state conservation policy shapes how national parks impact upon local resource access and use, older political economic inequalities in forest villages build on such policies to influence how management affects the livelihoods of poor households.

A GH receptor antisense oligonucleotide inhibits hepatic GH receptor expression, lGF-I production and body weight gain in normal mice

Diabetic retinopathy and acromegaly are diseases associated with excess action of GH and its effector IGF-1, and there is a need for improved therapies. We have designed all optimised 2'-O-(2-methoxyethyl)-modified phosphorothioate oligodeoxynucleotide, ATL 227446, and demonstrated its ability to Suppress GH receptor mRNA in vitro. Subcutaneous injections of ATL 227446 reduced GH receptor mRNA levels, GH binding activity and serum IGF-1 levels in mice after seven days of closing. The reduction in serum IGF-1 could be sustained for over tell weeks of dosing at therapeutically relevant levels, during which there was also a significant decrease in body weight gain in antisense-treated mice relative to saline and mismatch control-treated mice. The findings indicate that administration of an antisense oligonucleotide to the GH receptor may be applicable to human diseases in which suppression of GH action provides therapeutic benefit.

Animal production potential of some new Leucaena accessions in the Markham valley, Papua New Guinea

Signal grass pastures were oversown with four Leucaena spp. planted in hedgerows and evaluated for their agronomic productivity and ability to support steer liveweight gains. Each Leucaena sp. (L. leucocephala, L. pallida, L colli. nst. i., L. trichandra) was planted as seedlings into two I ha paddocks in rows 5 m apart, with I m spacing between trees. Cattle were rotationally grazed on the 2 replicates of each species, as well as on two I ha paddocks of a signal grass on y (Brachiaria decumbens) control, over a 243-day period at a stocking rate of 3 steers/ha. Mean presentation yield and herbage allowance of the Leucaena accessions over the grazing period were highest for L pallida (1100 kg/ha and 0.8 kg DM/kg LW, respectively), followed by L. leucocephala (700 kg/ha and 0.5 kg DM/kg LW), L. collinsii (700 kg/ha and 0.4 kg DM/kg LW) and L. trichandra (300 kg/ha and 0.2 kg DM/kg LW). Despite only moderate presentation yields and herbage allowances, steers grazing L. leucocephala and L. collinsii accessions produced the highest mean liveweight gains (LWG) of 0. and 0.56 kg/hd/d, respectively. While L. pallida produced the highest DM yields, it supported the lowest LWG of 0.36 kg/hd/d. The mean LWGs of steers grazing L. trichandra and the control (grass only) treatments were similar at 0.48 kg/ hd/d. The possible reasons for the differences in steer performance on the different Leucaena accessions are discussed.

Neuromuscular-skeletal constraints on the acquisition of skill in a discrete torque production task

The organisation of the human neuromuscular-skeletal system allows an extremely wide variety of actions to be performed, often with great dexterity. Adaptations associated with skill acquisition occur at all levels of the neuromuscular-skeletal system although all neural adaptations are inevitably constrained by the organisation of the actuating apparatus (muscles and bones). We quantified the extent to which skill acquisition in an isometric task set is influenced by the mechanical properties of the muscles used to produce the required actions. Initial performance was greatly dependent upon the specific combination of torques required in each variant of the experimental task. Five consecutive days of practice improved the performance to a similar degree across eight actions despite differences in the torques required about the elbow and forearm. The proportional improvement in performance was also similar when the actions were performed at either 20 or 40% of participants' maximum voluntary torque capacity. The skill acquired during practice was successfully extrapolated to variants of the task requiring more torque than that required during practice. We conclude that while the extent to which skill can be acquired in isometric actions is independent of the specific combination of joint torques required for target acquisition, the nature of the kinetic adaptations leading to the performance improvement in isometric actions is influenced by the neural and mechanical properties of the actuating muscles.

Production of triploid Kuruma shrimp, Marsupenaeus (Penaeus) japonicus (Bate) nauplii through inhibition of polar body I, or polar body I and II extrusion using 6-dimethylaminopurine

This study investigated the chromosome ploidy level of Marsupenaeus (Penaeus) japonicus (Bate) non-viable (unhatched) embryos and nauplii after exposure to 6-dimethylaminopurine (6-DMAP), timed to stop either polar body (PB) I, or PBI and II extrusion. Embryos from eight separate families or spawnings were exposed to 150 or 200 mu M 6-DMAP from 1- to 3-min post-spawning detection (psd) for a 4- to 5-min duration (timed to stop PBI extrusion). Separate aliquots of embryos from five of the same spawnings were also exposed to 200 mu M of 6-DMAP from 1- to 3-min psd for a 16-min duration (timed to stop both PBI and II extrusion). For one spawning, a third aliquot of embryos was exposed to 400 p M of 6-DMAP from 1- to 3-min psd for a 16-min duration (timed to stop both PBI and II extrusion). At 18-h psd, non-viable embryo and nauplii samples were taken separately for fluorescent activated cell sorting (FACS). FACS revealed that there were diploids and triploids among all treated non-viable embryos and nauplii. All control non-viable embryos and nauplii were diploid. Percentages of triploid induction for the 4- to 5-min and 16-min durations were not significantly different (P > 0.05). Additionally, no difference was found in the triploidy level of nonviable embryos compared to nauplii in these treatments. The percentage of triploid embryos and nauplii when exposed to 6-DMAP for a 4- to 5-min duration ranged from 29.57% to 99.23% (average 55.28 +/- 5.45%) and from 5.60% to 98.85% (average 46.70 +/- 7.20%), respectively. The percentage of triploid embryos and nauplii when exposed to 6-DMAP for a 16-min duration ranged from 11.71% to 98.96% (average 52.49 +/- 11.00%) and from 47.5% to 99.24% (average 79.38 +/- 5.24%), respectively. To our knowledge, this is the first documentation of successful PBI or PBI and II inhibition in shrimp. This study conclusively shows that treatment of M. japonicus embryos with 6-DMAP at 1- to 3-min pscl for either a 4- to 5-min duration (timed to stop PBl extrusion) or 16-min duration (timed to stop both PBI and II extrusion) results in viable triploid nauplii. (c) 2006 Elsevier B.V. All rights reserved.

Speech production in Parkinson's disease: I. An electropalatographic investigation of tongue-palate contact patterns

Previous studies have indicated that consonant imprecision in Parkinson's disease (PD) may result from a reduction in the amplitude of lingual movements or articulatory undershoot. While this has been postulated, direct measurement of the tongue's contact with the hard palate during speech production has not been undertaken. Therefore, the present study aimed to use electropalatography (EPG) to determine the exact nature of tongue-palate contact in a group of individuals with PD and consonant imprecision (n=9). Furthermore, the current investigation also aimed to compare the results of the participants with PD to a group of aged (n=7) and young (n=8) control speakers to determine the relative contribution of ageing of the lingual musculature to any articulatory deficits noted. Participants were required to read aloud the phrase 'I saw a ___ today' with the artificial palate in-situ. Target words included the consonants /l/, /s/ and /t/ in initial position in both the /i/ and /a/ vowel environments. Phonetic transcription of phoneme productions and description of error types was completed. Furthermore, representative frames of contact were employed to describe the features of tongue-palate contact and to calculate spatial palatal indices. Results of the perceptual investigation revealed that perceived undershooting of articulatory targets distinguished the participant group with PD from the control groups. However, objective EPG assessment indicated that undershooting of the target consonant was not the cause of the perceived articulatory errors. It is, therefore, possible that reduced pressure of tongue contact with the hard palate, sub-lingual deficits or impaired articulatory timing resulted in the perceived undershooting of the target consonants.

Speech production in Parkinson's disease: II. Acoustic and electropalatographic investigation of sentence, word and segment durations

Previous investigations employing electropalatography (EPG) have identified articulatory timing deficits in individuals with acquired dysarthria. However, this technology is yet to be applied to the articulatory timing disturbance present in Parkinson's disease (PD). As a result, the current investigation aimed to use EPG to comprehensively examine the temporal aspects of articulation in a group of nine individuals with PD at sentence, word and segment level. This investigation followed on from a prior study (McAuliffe, Ward and Murdoch) and similarly, aimed to compare the results of the participants with PD to a group of aged (n=7) and young controls (n=8) to determine if ageing contributed to any articulatory timing deficits observed. Participants were required to read aloud the phrase I saw a ___ today'' with the EPG palate in-situ. Target words included the consonants /1/, /s/ and /t/ in initial position in both the /i/ and /a/ vowel environments. Perceptual investigation of speech rate was conducted in addition to objective measurement of sentence, word and segment duration. Segment durations included the total segment length and duration of the approach, closure/constriction and release phases of EPG consonant production. Results of the present study revealed impaired speech rate, perceptually, in the group with PD. However, this was not confirmed objectively. Electropalatographic investigation of segment durations indicated that, in general, the group with PD demonstrated segment durations consistent with the control groups. Only one significant difference was noted, with the group with PD exhibiting significantly increased duration of the release phase for /1a/ when compared to both the control groups. It is, therefore, possible that EPG failed to detect lingual movement impairment as it does not measure the complete tongue movement towards and away from the hard palate. Furthermore, the contribution of individual variation to the present findings should not be overlooked.

Top-down influences on lexical selection during spoken word production: A 4T fMRI investigation of refractory effects in picture naming

Spoken word production is assumed to involve stages of processing in which activation spreads through layers of units comprising lexical-conceptual knowledge and their corresponding phonological word forms. Using high-field (4T) functional magnetic resonance imagine (fMRI), we assessed whether the relationship between these stages is strictly serial or involves cascaded-interactive processing, and whether central (decision/control) processing mechanisms are involved in lexical selection. Participants performed the competitor priming paradigm in which distractor words, named from a definition and semantically related to a subsequently presented target picture, slow picture-naming latency compared to that with unrelated words. The paradigm intersperses two trials between the definition and the picture to be named, temporally separating activation in the word perception and production networks. Priming semantic competitors of target picture names significantly increased activation in the left posterior temporal cortex, and to a lesser extent the left middle temporal cortex, consistent with the predictions of cascaded-interactive models of lexical access. In addition, extensive activation was detected in the anterior cingulate and pars orbitalis of the inferior frontal gyrus. The findings indicate that lexical selection during competitor priming is biased by top-down mechanisms to reverse associations between primed distractor words and target pictures to select words that meet the current goal of speech.

The role of CD4(+) and CD8(+) T cells on antibody production by murine Peyer's patch cells following mucosal presentation of Actinomyces viscosus

The aim of this study was to determine the role of CD4 and CD8 cells on specific antibody production by murine Peyer's patch (PP) cells after oral immunization with Actinomyces viscosus in mice. Female DBA/2 mice were orally immunized with three low doses of heat-killed A. viscosus. Sham-immunized mice served as a control group. Mice were depleted of CD4 or CD8 cells by intraperitoneal injection of anti-CD4 or anti-CD8 antibodies daily for 3 days before oral immunization. One week after the last oral immunization, PPs were removed and cell suspensions were cultured with A. viscosus. Specific antibody production in the culture supernatants was assessed by enzyme-linked immunosorbent assay. The results showed that oral immunization with A. viscosus induced a predominant specific immunoglobulin A (IgA) response by PP cells and, to a lesser extent, IgM antibodies. Depletion of CD4 but not CD8 cells suppressed the production of specific antibodies. These results suggest that oral immunization with low doses of A. viscosus may induce the production of specific antibodies by murine PP cells in a CD4-cell-dependent fashion.

In vitro production of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus

Naturally occurring insect viruses are a promising means of intentionally causing disease in insects but they do not compete successfully with synthetic chemicals in the commercial marketplace. Furthermore, their use for pest control is still restricted. One factor preventing the development of baculoviruses as effective biopesticides is concern over the production issue. In vitro instability during propagation of these viruses in suspension cells is the major limitation to the in vitro production ofbaculoviruses in cell cultures. In this study, an isolated baculovirus (HaSNPV) was cultivated using serial passaging in a suspension cell culture. The results show a reduction in the occlusion body production during six passages, due to the passage effect. However the purification of an HaSNPV clone suggested better stability. A simple method used in this work for the serial passaging of this virus is discussed.

'Looking good, feeling better': Consumption, Control and the Aesthetics of Exclusivity in the Service Encounter

The majority of ‘service’ literature has focused on the production side of service work (i.e. employees and management), while treating the role of the customer and/or consumer as secondary (Korczynski and Ott, 2004). Those authors who have addressed the role consumption plays in shaping and maintaining individuals' self- identity have tended to overemphasize the dominance of consumer culture in shaping ‘our consciousness’ (Ritzer, 1999), with little in the way of empirical evidence to support these assertions. This paper develops the conceptualization of service work and consumer culture literature, by placing more emphasis on the customer in the service encounter. Using an ethnographic study of a ‘high class’ department store, this paper addresses employee and customer identity and the nature of managerial, employee and customer control within this ‘exclusive’ context. Of particular interest is how employees and customer’s ‘embody’ this control. Using Bourdieu’s (1986) conception of class and habitus, the concept of exclusivity goes beyond the management /service worker dyad by providing a means of investigating identity control by the organization over both customers and service workers. However, an organization’s exclusivity is not a closed normative pursuit of control, and shows this enterprise is part of a contested terrain, while revealing the ambiguity and ‘openness’ of control practices and pursuits. In order to uphold the ideal of exclusivity, management, service workers and customers must all engage in a precarious quest for establishing and maintaining a sense of control and/or identity. This paper demonstrates the continuing contradiction between bureaucratic practices of control and consumer culture, and highlights the need for research that investigates the context -dependent nature of control in service-related and consumer studies.