Understanding cell culture dynamics: a tool for defining protocol parameters for improved processes and efficient manufacturing using human embryonic stem cells.

Standardization is crucial when culturing cells including human embryonic stem cells (hESCs) which are valuable for therapy development and disease modeling. Inherent issues regarding reproducibility of protocols are problematic as they hinder translation to good manufacturing practice (GMP), thus reducing clinical efficacy and uptake. Pluripotent cultures require standardization to ensure that input material is consistent prior to differentiation, as inconsistency of input cells creates end-product variation. To improve protocols, developers first must understand the cells they are working with and their related culture dynamics. This innovative work highlights key conditions required for optimized and cost-effective bioprocesses compared to generic protocols typically implemented. This entailed investigating conditions affecting growth, metabolism, and phenotype dynamics to ensure cell quality is appropriate for use. Results revealed critical process parameters (CPPs) including feeding regime and seeding density impact critical quality attributes (CQAs) including specific metabolic rate (SMR) and specific growth rate (SGR). This implied that process understanding, and control is essential to maintain key cell characteristics, reduce process variation and retain CQAs. Examination of cell dynamics and CPPs permitted the formation of a defined protocol for culturing H9 hESCs. The authors recommend that H9 seeding densities of 20,000 cells/cm2, four-day cultures or three-day cultures following a recovery passage from cryopreservation and 100% medium exchange after 48 hours are optimal. These parameters gave ~SGR of 0.018 hour-1 ± 1.5x10-3 over three days and cell viabilities ≥95%±0.4, while producing cells which highly expressed pluripotent and proliferation markers, Oct3/4 (>99% positive) and Ki-67 (>99% positive).

Oil sands tailings ponds harbour a small core prokaryotic microbiome and diverse accessory communities.

Oil sands tailings ponds store the waste slurry generated by extracting bitumen from surface-mined oil (tar) sands ores. The ponds support diverse microbial communities involved in element cycling, greenhouse gas production, and hydrocarbon biodegradation that influence pond management and their environmental footprint. Since previous reports indicate that there are similar microbial metabolic functions amongst ponds, analogous microbiomes may be expected but ponds actually harbour distinct communities. Partial 16S rRNA gene pyrotag sequences from 95 samples were obtained from six ponds managed by three operators. From these we discerned a core prokaryotic microbiome, a subset of microbes shared amongst different samples, defined as operational taxonomic units (OTUs) at the lowest taxonomic level identifiable in individual ponds and pooled pond datatsets. Of the ∼1500-2700 OTUs detected per pond, 4-10 OTUs were shared among ≥75% of the samples per pond, but these few OTUs represented 39-54% of the ponds' sequence reads. Only 2-5 OTUs were shared by the majority of samples from all ponds. Thus the prokaryotic communities within these ponds consist of a few core taxa and numerous accessory members that likely afford resiliency and functional redundancy including roles in iron-, nitrogen- and sulfur-cycling, syntrophy, fermentation, and methanogenesis.

Detecting somatic mosaicism: considerations and clinical implications.

Human disease is rarely a matter of all or nothing; variable expressivity is generally observed. Part of this variability is explained by somatic mosaicism, which can arise by a myriad of genetic alterations. These can take place at any stage of development, possibly leading to unusual features visible at birth, but can also occur later in life, conceivably leading to cancer. Previously, detection of somatic mosaicism was extremely challenging, as many gold standard tests lacked the necessary resolution. However, with the advances in high-throughput sequencing, mosaicism is being detected more frequently and at lower levels. This raises the issue of normal variation within each individual vs mosaicism leading to disease, and how to distinguish between the two. In this article, we will define somatic mosaicism with a brief overview of its main mechanisms in concrete clinical examples, discuss the impact of next-generation sequencing technologies in its detection, and expand on the clinical implications associated with a discovery of somatic mosaicism in the clinic.

Effects of HIV status and antiretroviral therapy on blood pressure.

OBJECTIVE: High blood pressure is a major risk factor for cardiovascular disease and concerns have been raised over its possible association with antiretroviral drugs. The objective of this study was to explore the associations among blood pressure, HIV status and two predefined highly active antiretroviral therapy (HAART) regimens: treatment with and without nonnucleoside reverse transcriptase inhibitors (NNRTIs) (NNRTI- and non-NNRTI-based HAART). METHOD: A cross-sectional survey was conducted among 612 adults attending the Sexual Health Outpatient Department at St Mary's NHS Hospital Trust, London. RESULTS: HIV-infected patients treated with NNRTIs had a blood pressure that was 4.6/4.2 mmHg higher than those who were HIV positive but treatment naïve. The diastolic difference remained statistically significant after adjusting for potential confounders of this association (2.4 mmHg; P=0.03). There was no difference in blood pressure between those treated with non-NNRTI-based regimens and those who were HIV positive but treatment naïve. CONCLUSION: NNRTIs may be associated with an increase in blood pressure. Pending further more robust evidence from randomized clinical trials it would be prudent for clinicians to monitor blood pressure in all HIV-infected patients, particularly after initiating treatment with NNRTIs, and to commence antihypertensive therapy whenever appropriate.

Psychotherapy with brain injury survivors: an investigation of the challenges encountered by clinicians and their modifications to therapeutic practice.

OBJECTIVE: The aim of this study is to elicit practitioners' views and experiences of the challenges to forming a therapeutic alliance with brain injury survivors, with a view to informing current psychotherapeutic practice. METHODS: The present research utilised the data-display method, a qualitative technique, to examine the questionnaire responses of 21 psychologists who provide forms of psychotherapy for brain injury survivors at rehabilitation units in the UK. An anonymous postal return questionnaire was used for data collection. RESULTS: The main challenges to forming a working alliance comprised a range of cognitive, behavioural and emotional sequelae. A combination of educational, psychosocial and cognitive strategies were identified as being most effective in addressing the challenges encountered. CONCLUSIONS: A qualitative research approach has proved useful in identifying challenges to the formation of a working alliance and also the modifications to psychotherapeutic practice these challenges have engendered.

In vitro and In vivo inhibition of LPS-stimulated tumor necrosis factor-alpha secretion by the gallotannin beta-D-pentagalloylglucose.

The naturally occurring gallotannin beta-D-pentagalloylglucose (beta-PGG) decreases tumor necrosis factor-alpha (TNF-alpha) output from human peripheral blood mononucleocytes exposed to lipopolysaccharide (LPS) by as much as 90% (vs control) at approximately 5 microM concentration. A qualitatively similar but less pronounced effect ( approximately 50% decrease) was observed in the serum of rats dosed with both LPS and beta-PGG. These results may have relevance to therapies that target disease states characterized by an overproduction of TNF-alpha.

Differences in attitudes regarding teaching and research orientations between allied health faculty and administrators in academic health centers.

The congruence between faculty's and administrators' attitudes toward teaching, research, and the rewards associated with them has implications for the practices the groups follow and for the decisions they make related to promotion, tenure, and merit. This article reports the findings from a national study of allied health faculty and administrators at public academic health centers regarding their attitudes toward teaching, research, and the reward systems related to both. The administrators and faculty at the participating academic health centers did not differ in their views regarding research orientation and rewards related to research. The faculty, including tenured and tenure-track, and department chairs indicated a higher teaching orientation than did the deans, while tenure-track faculty expressed less belief that rewards influence teaching than did tenured faculty, department chairs, and deans.

Misconceptions about brain injury among the general public and non-expert health professionals: an exploratory study.

The purpose of this pilot study was to investigate the lack of knowledge and misconceptions concerning brain injury, as perceived by those with experience of the condition. Using a qualitative research method, 19 semi-structured interviews were conducted with brain-injured individuals, caregivers and professionals who provide social rehabilitation after brain injury. Interviews were analysed using Interpretative Phenomenological Analysis. According to participants, inaccurate and inadequate knowledge about brain injury is common among the general public and among health professionals without expertise in the field of brain injury. The major themes that emerged from the analysis were: inaccurate beliefs about recovery time and possible extent of recovery from brain injury; lack of awareness of the diversity ofproblems it can cause, particularly the existence of behavioural and cognitive sequelae; misconceptions about the capabilities of brain-injured people depending on the visibility or invisibility of their disability: and misidentification of brain-injured individuals as mentally ill or learning disabled. Results are discussed in terms of a theory of illness cognition. Posibilities for further research are discussed, and it is concluded that the results of this study could help guide future information provision to all who may come into contact with brain injury.

Attachment disorders: review and current status.

Developmental research on attachment has flourished in the past 15 years (C. H. Zeanah, 1996). However, there has been relatively scant empirical investigation of disorders in attachment. In this article, the pertinent developmental research on the attachment cycle is delineated and the current status of disordered attachment is examined. A particular focus is given to the conceptualization of the most severe form of disordered attachment, Reactive Attachment Disorder.

Cardiovascular risk: its assessment in clinical practice.

Cardiovascular disease is one of the major causes of mortality and morbidity in Western industrialised countries. Many factors impact on the likelihood of an individual suffering a stroke or heart attack. One of the greatest challenges facing clinicians today is how best to identify those individuals at increased risk, to prevent them becoming tomorrow's patients. Risk assessment tools can provide useful frameworks to support the identification of individuals who may benefit from therapeutic intervention; however, such tools should be used with care, as they do not include all the factors that contribute to future disease risk, and they are subject to an age bias. Nonetheless, they provide a systematic, evidence-based approach to the delivery of preventative healthcare services.

Hypertension awareness, detection and treatment in a university community: results of a worksite screening.

This study aimed to assess knowledge levels, explore the extent of undiagnosed hypertension, record previous screening activity and monitor the outcomes of previous hypertension treatment. To this end, information was collected from volunteer members of a university community, by means of a self-report questionnaire, about their personal characteristics, knowledge and experience of hypertension and stroke, and previous blood pressure measurements. In addition, their current blood pressure was recorded. A total of 653 staff and students completed the questionnaire and had their blood pressure measured. Analysis revealed that 82% were normotensive (diastolic blood pressure <90 mmHg). Hypertension was significantly associated with age, self-reported excess weight P<0.001) and marginally with self-reported non-healthy eating (P=0.06). Of the volunteers, 57% could not provide an accurate definition of a stroke. Knowledge levels were significantly and positively related to experience of stroke, healthy eating, not smoking and a recent blood pressure check. Of the respondents, 30% stated that they had not had their blood pressure measured in the previous two years. 51% of known hypertensives were not controlled. 68% of volunteers with diastolic blood pressure >89 mmHg were previously unaware of a potential hypertension problem. Hypertension rates amongst the university volunteers are higher than those recently recorded from a population sample. Scope exists for increasing knowledge and awareness, and for raising both screening rates and treatment outcomes. Improvements in these areas are required if current public health targets for heart disease and stroke are to be achieved. Worksite screening programmes can contribute to this endeavour.

Representation: a call to action for allied health professionals.

The Coalition of Allied Health Leadership (CAHL) Representation Project committee examined the representation of allied health professionals in political and other policy-making groups and found it both fragmented and lacking. The benefits to individuals participating in such groups, as well as to the allied health profession as a whole and to the groups themselves, are described. Individuals are urged to participate, and the means to do so are presented.

Early indication of emergence from vegetative state derived from assessments with the SMART--a preliminary report.

An explanatory analysis of data from serial assessments of 30 patients with a diagnosis of vegetative state (persistent vegetative state) was carried out. The data were gathered using the Sensory Modality Assessment and Rehabilitation Technique (SMART) and the Western Neuro Sensory Stimulation Profile (WNSSP) on the same day in the week post-admission and, thereafter, at 2 monthly intervals. Seven patients emerged from vegetative state during the study, a further six emerged later (by time of the analysis) and the rest remained in vegetative state. Recovery scores were calculated for each measure by subtracting the total score at any one assessment from the total score for the subsequent assessment; the largest recovery scores, shown by each patient, were noted. It was found that those who emerged later could be differentiated mathematically from those who did not emerge, using largest recovery score data from the SMART. This research supports previous

Chromobacteriosis in a Chinese red panda (Ailurus fulgens styani).

An adult Chinese red panda (Ailurus fulgens styani) transported by airplane from Florida to a North Dakota zoo died 1 week after arrival. Grossly, an interscapular abscess, subcutaneous inflammation, lymphadenitis, and pulmonary abscesses were observed. Microscopic findings included necrotizing inflammation in liver, lung, lymph node, and spleen. Chromobacterium violaceum was cultured from the interscapular abscess, liver, lung, and spleen and was injected into Swiss Webster mice. These mice died 18 hours postinoculation, and C. violaceum was cultured from liver, lung, and spleen. Chromobacterium violaceum is a sporadically reported but highly virulent pathogenic bacterium of both animals and humans typically found as a soil and water inhabitant of tropical and subtropical regions.

Scholarship and research of allied health deans and directors.

In 1997, a questionnaire was sent to 141 allied health deans or directors to: 1) determine current research and scholarship, 2) ascertain institutional expectations regarding scholarly activities, 3) characterize the institution's research environment, 4) quantitate scholarship productivity by point assessment, and 5) compare scholarship of administrators from research universities with that of those at non-research four-year colleges and universities. Seventy-three (52%) responded. Measures of research productivity included time spent in research, numbers of publications and presentations, and grantsmanship. While spending two thirds of their time in administrative duties, many deans and directors were actively involved in scholarship. Since 1990, they had, as a group, produced 855 publications, made 1,348 presentations, and been awarded $38.3 million in total funding. They did not differ significantly by employing institution (a research institution or a non-research institution).

Listeria monocytogenes infection and activation of human brain microvascular endothelial cells.

Listeria monocytogenes invasion of human brain microvascular endothelial cells (BMEC) and its role as a stimulus for endothelial cell activation were studied. Binding and invasion of intact BMEC monolayers were independent of the L. monocytogenes inlAB invasion locus. Cytochalasin D abrogated invasion of BMEC, whereas genistein effected only a 53% decrease in invasion, indicating a requirement for rearrangement of actin microfilaments but less dependence on tyrosine kinase activity. L. monocytogenes stimulated surface expression of E-selectin, ICAM-1, and to a lesser extent, VCAM-1, whereas L. monocytogenes prfA- and Deltahly mutants were severely compromised in this respect. Other experiments showed that BMEC infection stimulated monocyte and neutrophil adhesion and that CD18-mediated binding was the predominant mechanism for neutrophil adhesion to infected BMEC under static conditions. These data suggest that invasion of BMEC is a mechanism for triggering inflammation and leukocyte recruitment into the central nervous system during bacterial meningitis.

Cardiac pacing in Canada in 1998: working towards optimal pacing therapy. Canadian Working Group on Cardiac Pacing.

The Canadian Working Group on Cardiac Pacing (CWGCP) was formed in 1996 with the primary goal of promoting optimal pacing therapy in Canada. In 1997, the CWGCP conducted a survey of pacing practices across Canada. Ninety-two of 125 implanting programs (74%) responded. Implant rates vary by province--from 39 per 100,00 population in Ontario to 63 per 100,000 population in Nova Scotia and Prince Edward Island. Variations in regional implant rates persist even after correcting for the age of the population. Physiological pacing was used for 35% of all implants in Canada in 1996/97. There were marked differences across Canada in the mode of pacing selected. In western Canada, 39.5% of pacing systems implanted were physiological compared with 18.2% in Atlantic Canada and 29% in Quebec. There were also differences in follow-up practices. Approximately 40% of centres follow patients with single chamber pacemakers annually, whereas most other centres still follow these patients every six months. Economic constraints, the size of pacing programs and the involvement of committed pacing physicians are factors that may influence the regional differences in cardiac pacing across Canada.