A three-dimensional model of primary bovine endometrium using an electrospun scaffold.

Endometrial stromal and epithelial cell function is typically studied in vitro using standard two-dimensional monocultures, but these cultures fail to reflect the complex three-dimensional (3D) architecture of tissue. A 3D model of bovine endometrium that reflects the architectural arrangement of in vivo tissue would beneficially assist the study of tissue function. An electrospun polyglycolide (PGA) scaffold was selected to grow a 3D model of primary bovine endometrial epithelial and stromal cells, that reflects the architecture of the endometrium for the study of pathophysiology. Electrospun scaffolds were seeded with stromal and epithelial cells, and growth was assessed using histological techniques. Prostaglandin E2 and prostaglandin F2α responsiveness of endometrial scaffold constructs was tested using oxytocin plus arachidonic acid (OT + AA) or lipopolysaccharide (LPS). Stromal and epithelial cells growing on the electrospun scaffold had an architectural arrangement that mimicked whole tissue, deposited fibronectin, had appropriate expression of vimentin and cytokeratin and were responsive to OT + AA and LPS, as measured by prostaglandin accumulation. In conclusion, a functional 3D model of stromal and epithelial cells was developed using a PGA electrospun scaffold which may be used to study endometrial pathophysiology.

The experience of acupuncture care from the perspective of people with chronic low back pain: a grounded theory study.

OBJECTIVES: To explore the experience of acupuncture care from the perspective of people with chronic low back pain. METHODS: A grounded theory, qualitative study using indepth interviews was undertaken with 11 people who had received acupuncture care for chronic low back pain. Interviews were audio-recorded and transcribed verbatim. Initial, focused and theoretical coding were used to identify a core category and several other categories. RESULTS: The core category, Reclaiming Control, was related to the sense of well-being experienced by most of the participants as a result of undergoing acupuncture care. The other categories were Gaining Sanctuary, Gaining Trust and Working Together. These categories reflected the processes of participants entering an aesthetically appealing, calm and relaxing space; developing confidence in the acupuncturists' ability to care for them; and negotiating strategies and sharing decision-making with acupuncturists about their care. CONCLUSIONS: Clinicians using acupuncture as a management strategy for low back pain may elicit an immediate sense of calmness in patients with subsequent well-being benefits. A sense of calmness may also be enhanced through providing a relaxing physical environment. The transactional and interpersonal processes of establishing trust and rapport, and shared decision-making are important for clients. Shared decision-making can be improved by clinicians carefully considering the explanation of issues and plans to a particular patient, especially by developing an understanding of each patient's worldview and using language that the suits each individual's healthcare orientation. Our findings should be interpreted cautiously as the results may be generalisable only to private practice in affluent countries, and whether the findings are representative beyond this setting remains unclear.

Elucidating the relative roles of ammonia oxidizing and heterotrophic bacteria during the biotransformation of 17α-Ethinylestradiol and Trimethoprim.

The biological fate of 17α-ethinylestradiol (EE2; 500 ng/L to 1 mg/L) and trimethoprim (TMP; 1 µg/L to 1 mg/L) was evaluated with flow through reactors containing an ammonia oxidizing bacterial (AOB) culture, two enriched heterotrophic cultures devoid of nitrifier activity, and nitrifying activated sludge (NAS) cultures. AOBs biotransformed EE2 but not TMP, whereas heterotrophs mineralized EE2, biotransformed TMP, and mineralized EE2-derived metabolites generated by AOBs. Kinetic bioassays showed that AOBs biotransformed EE2 five times faster than heterotrophs. The basal expression of heterotrophic dioxygenase enzymes was sufficient to achieve the high degree of transformation observed at EE2 and TMP concentrations ≤ 1 mg/L, and enhanced enzyme expression was not necessary. The importance of AOBs in removing EE2 and TMP was evaluated further by performing NAS experiments at lower feed concentrations (500-1000 ng/L). EE2 removal slowed markedly after AOBs were inhibited, while TMP removal was not affected by AOB inhibition. Two key EE2 metabolites formed by AOB and heterotrophic laboratory-scale chemostats were also found in independent laboratory-scale mixed culture bioreactors; one of these, sulfo-EE2, was largely resistant to further biodegradation. AOBs and heterotrophs may cooperatively enhance the reliability of treatment systems where efficient removal of EE2 is desired.

Isokinetic strength training of lower limb muscles following acquired brain injury.

PRIMARY OBJECTIVE: To investigate the effectiveness of isokinetic strength training of ankle and knee muscles in adults with chronic acquired brain injury (ABI). RESEARCH DESIGN: Series of single case studies. METHODS: Twelve people with ABI participated in a 2.5-week baseline, 12-week intervention and a 4-week follow-up phase. INTERVENTION: Concentric isokinetic exercise, twice a week, for plantarflexors (PFs), dorsiflexors (DFs), knee flexors (KFs) and knee extensors (KEs). OUTCOMES: Peak torque and power at 60 and 90° s⁻¹, PFs and KFs tone at 60° s⁻¹, gait speed and timed chair rises. RESULTS: For single case analyses strength improvements were noted in 11/12 participants' PFs, 5/12 participants' DFs and 7/12 participants' KEs and KFs. Gait speed improved in 8/12 participants and chair rise time improved in 7/12 participants. PFs tone increased in three participants, KFs tone increased in six participants and three participants reported knee pain. For group analyses, peak torque of PFs and KEs, fast gait speed and timed chair rises demonstrated improvement (p < 0.05). CONCLUSIONS: Isokinetic strength training may be effective to improve lower limb muscle strength; however, care needs to be taken in selecting suitable candidates as some individuals reported knee pain with this intensive programme.

An isokinetic muscle strengthening program for adults with an acquired brain injury leads to meaningful improvements in physical function.

PRIMARY OBJECTIVE: To investigate if an isokinetic strength training programme for leg muscles lead to personally meaningful changes in adults with an acquired brain injury (ABI). RESEARCH DESIGN: A qualitative exploratory design. METHODS: Twelve people with ABI participated in pre- and post-intervention face-to-face interviews with open ended questions. Data were initially analysed using a case study research approach exploring individuals experiences and then cross case analysis to determine common themes for the group. INTERVENTION: Twelve-week isokinetic strength training programme for ankle and knee muscles. OUTCOMES: Participants perceived changes. RESULTS: Thematic analysis determined four main themes arising from the interviews; occupation, vitality, sense of self and personal interactions. Participants reported reductions in impairments as a response to the exercise programme and these changes led to increased function and participation in activities they valued. Also marked improvements in vitality were reported as well as increases in self-esteem and general well-being for many participants. CONCLUSIONS: An isokinetic strength training programme resulted in improvements in motor skills and functional abilities that were meaningful for the participants.

Acupuncturists' perspectives on outcome measures to evaluate acupuncture care for chronic low back pain.

OBJECTIVES: To examine Australian acupuncturists' perspectives of: chronic low back pain outcome domains; their use of outcome measures to assess chronic low back pain; and their attitudes and perceptions of barriers to using the existing measures to evaluate acupuncture care for chronic low back pain. DESIGN: A postal questionnaire was used in a cross-sectional survey. PARTICIPANTS: 359 randomly selected Australian acupuncturists. RESULTS: 139 questionnaires were returned (response rate: 38.7%). The respondents' demographic characteristics were similar to Australian Bureau of Statistics acupuncturist demographic data. Overall, acupuncturists endorsed a broad approach to assessing acupuncture care for chronic low back pain. However, they had typically used pain severity measures but generally did not use other types of measures. Acupuncturists tended to hold positive attitudes towards using outcome measures. The principal barriers acupuncturists perceived to using outcome measures concerned doubts about whether the concepts which underpin acupuncture practice had been explicitly articulated and whether the available measures capture the specific context and particular outcomes of acupuncture care for chronic low back pain. IMPLICATIONS: The findings of this study suggest that a broad range of outcome measures are required to adequately assess chronic low back pain acupuncture care outcomes. While numerous outcome measures have been developed that are relevant to chronic low back pain care, whether these measures are appropriate for use by acupuncturists is unclear. Further studies are warranted to explore if established outcome measures are useful to evaluate chronic low back acupuncture care.

Falls and injury prevention should be part of every stroke rehabilitation plan.

OBJECTIVE: To evaluate falls incidence, circumstances and consequences in people who return home after stroke rehabilitation, so that appropriate falls and injury prevention strategies can be developed. DESIGN: Prospective cohort study. SETTING: Community. SUBJECTS: Fifty-six subjects with stroke who were participating in a rehabilitation programme and returning to live in a community setting completed the study. MAIN MEASURES: Subjects completed a prospective falls diary for six months after discharge from rehabilitation, and were interviewed after falls. Physical function was measured by the Berg Balance Scale (BBS) and the Functional Independence Measure (FIM). RESULTS: Forty-six per cent of people (26/56) fell, with most falls (63/103 falls) occurring in the two months after discharge from rehabilitation. One subject had 37 similar falls and these falls were excluded from further analysis. Falls occurred more often indoors (50/66), during the day (46/66) and towards the paretic side (25/66). People required assistance to get up after 25 falls (38%) and 36 falls (55%) resulted in an injury. People sought professional health care after only 16 falls, and activity was restricted after 29 falls (44%). The Berg Balance Scale and Functional Independence Measure scores were lower in people who had longer lies after a fall, and who restricted their activity after a fall (p < 0.05). Lower physical function scores were also associated with falling in the morning, wearing multifocal glasses at the time of a fall, and injurious falls (p < 0.05). CONCLUSION: Falls are common when people return home after stroke. Of concern are the small number seeking health professionals' assistance after a fall, the high proportion restricting their activity as a result of a fall and the number of falls occurring towards the paretic side.

A single amino acid is critical for the expression of B-cell epitopes on the helicase domain of the pestivirus NS3 protein.

Truncated NS3 proteins, expressed by recombinant baculoviruses, were used to investigate the location of conserved B-cell epitopes on this non-structural bovine viral diarrhoea virus (BVDV) protein. A goat anti-pestivirus antiserum, and a panel of anti-NS3 monoclonal antibodies, including the BVDV-1 specific antibody P1D8, were used to verify the presence or absence of the epitopes. Interestingly, the monoclonal antibodies reacted only with the truncated protein encompassing the helicase domain of NS3. Expression of the B-cell epitopes was dependent on, but not within, a 57 amino acid sequence at the carboxy-terminal end of this protein, supporting observations that these conserved epitopes are conformational in nature. A comparison of deduced amino acid sequences of the helicase domain from BVDV-1, BVDV-2, BDV and CSFV isolates highlighted a single amino acid that appeared to be unique to P1D8-reactive BVDV-1 isolates. Site-directed mutagenesis studies confirmed that this amino acid is critical for the expression of the BVDV-1 specific NS3 epitope recognised by the P1D8 monoclonal antibody. Surprisingly, the amino acid was also important for an epitope recognised by two group-specific monoclonal antibodies, P1H11 and P4A11. Protein modelling studies, based on the structure of the hepatitis C NS3 helicase domain, indicated that this amino acid occupies a prominent position on the surface of the protein.

A stable cell line with a proportion of cells persistently infected with bovine viral diarrhoea virus.

Bovine turbinate (BTu) and lamb testis (LT) cell lines persistently infected with bovine viral diarrhoea virus (BVDV) arose as a result of a single change of medium containing commercial foetal calf serum. Infected cells comprise 30% and 50% respectively, of the total cell population, determined by immunohistochemical staining. The ratio of positive cells has remained unchanged during successive passages. Characterization of the persistently-infected BTu cells (BTuI) showed that only full length viral RNA was detected by northern blot hybridisation, indicating that DI particles were not involved. Secreted and intracellular virus from these cells was fully infectious for fresh BTu and LT cells. The BTuI cell line was fully permissive for a cytopathic BVDV isolate and a bovine herpesvirus, but non-permissive for two non-cytopathic BVDV isolates. Attempts to induce the permissive state in the BVDV-negative cells of the BTuI culture by treatment with actinomycin D and 5'-aza-cytidine failed. These cells provide a convenient model to study aspects of BVDV pathogenesis and replication.

Mutational analysis of the NH2-terminal arms of the trp repressor indicates a multifunctional domain.

The NH2-terminal arms of the Escherichia coli trp repressor have been implicated in three functions: formation of repressor-operator complexes via association with non-operator DNA; stabilization of repressor oligomers bound to DNA; and oligomerization of the aporepressor in the absence of DNA. To begin to examine the structural aspects of the arms that are responsible for these varied activities, we generated an extensive set of deletion and substitution mutants and measured the activities of these mutants in vivo using reporter gene fusions. Deletion of any part of the arms resulted in a significant decrease in repressor activity at both the trp and the trpR operons. Positions 4, 5 and 6 were the most sensitive to missense changes. Most substitutions at these positions resulted in repressors with less than 5% of the activity of the wild-type trp repressor. A large percentage of the missense mutants were more active than the wild-type repressor in medium containing tryptophan and less active in medium without tryptophan. This phenotype can be explained in terms of altered oligomerization of both the repressor and the aporepressor. Also, nine super-repressor mutants, resulting from substitutions clustered at both ends of the arms, were found. Our results support the hypothesis that the NH2-terminal arm of the trp repressor is a multifunctional domain and reveal structural components likely to be involved in the various functions.

Technology in education: what is appropriate for rural and remote allied health professionals?

Recent technological advances provide exciting opportunities for the delivery of education to rural and remote allied health professionals. Distance education modes can overcome barriers of distance and cost. However, the use of technologies may allow distance education to be more interactive for rural and remote professionals. Technologies that can be used are audioconferencing, videoconferencing, electronic mail (email), CD-ROM and the Internet. Combinations of any of these technologies and traditional forms of educational delivery are possible. The important objective is to create a learning environment that maximises interactivity and develops information literacy.

Insemination of cattle with semen from a bull transiently infected with pestivirus.

When 73 heifers (60 of which were seronegative to pestivirus) were inseminated with pestivirus-contaminated semen from a transiently infected bull, the conception rate to a single insemination was found to be normal (65 per cent). Only three animals became systemically infected, as determined by viraemia and seroconversion. Pestivirus was isolated from the reproductive tracts of two of these heifers when they were slaughtered 42 or 43 days after insemination. Although the initial incidence of infection was low, a cycle of secondary transmission occurred approximately 29 days after insemination, with a further eight heifers (all seronegative) becoming infected from one group of 11 seronegative and four seropositive animals.

The outcome of widespread use of semen from a bull persistently infected with pestivirus.

During the certification of the bulls at an artificial breeding centre for freedom from pestivirus infection, a single viraemic bull was identified, and further testing confirmed that it was persistently infected. The two-year-old bull was healthy and of similar bodyweight to its peers. Its semen was of normal quality on the basis of density, motility and morphological criteria. Approximately 600 doses of semen had been distributed for sire evaluation purposes to 97 dairy farms. An examination of the breeding records indicated a first service conception rate of 38 per cent. All but one of the 162 cows inseminated with the bull's semen were seropositive compared with 95 of 143 cows (66.4 per cent) inseminated with semen from other bulls. Virological studies of the 61 calves sired by the persistently infected bull revealed that two were persistently infected, but that the others were healthy and uninfected. It was concluded that the semen from this bull was a potential source of pestivirus infection for 'clean' herds.

Detection of hog cholera virus antigens in experimentally-infected pigs using an antigen-capture ELISA.

An antigen-capture ELISA was used to detect hog cholera virus (HCV) antigens in blood and tissues taken from pigs infected with 2 different strains of virus. Specific antigens were demonstrated in peripheral blood leucocytes (PBLs) and a wide range of tissue samples 4-6 days after infection of pigs with a moderate-high virulent HCV strain (Weybridge virus). Strong signal to noise (S/N) ratios were obtained in the ELISA for PBLs and lymphoid tissues such as spleen, tonsil and mesenteric lymph nodes at 5-7 days after infection with the Weybridge virus, S/N ratios varying between 8.1-19.7 for blood samples and 4.3-19.1 for spleen samples. High positive ELISA results were also obtained for duodenum and ileum samples (S/N ratios 10.3-18.6) taken from these pigs, reflecting severe pathological changes observed in the gut at post mortem. In contrast, the antigen-capture ELISA gave strong positive results for PBLs and spleen samples only at 7-9 days after infection of pigs with a low virulent strain of HCV (New South Wales virus). The ELISA S/N ratios averaged 9.5 for PBLs and 8.9 for spleen samples in these animals. Although virus isolation detected infection earlier in the infected pigs, the ELISA returned positive results on PBLs and spleen samples around the time all of the animals first showed typical signs of classical swine fever. The technique does not require tissue culture and takes less than 36 h to return a definitive result.