What factors influence the uptake of vaccinations amongst pregnant women following the Covid-19 pandemic: A qualitative study.

BACKGROUND: Pregnant women and their unborn babies are at increased risk from serious complications, hospitalisation and death from infectious diseases. Vaccinations for influenza (flu), pertussis (whooping cough) and Covid-19 are available for free for pregnant women in the UK, but uptake of these repeatedly remains low. This qualitative study aimed to explore how pregnant women feel about these vaccinations, and what factors influence the uptake of vaccinations amongst pregnant women since the onset of the Covid-19 pandemic. METHODS: Pregnant women were recruited via two participating hospitals in one geographic area of the UK, and via one community group offering support to pregnant women from ethnic minorities. Semi-structured interviews were conducted remotely using telephone, were anonymised and transcribed, and analysed using thematic analysis. FINDINGS: Interviews were conducted remotely with 43 pregnant women. The following themes were identified as influencing uptake of vaccinations amongst pregnant women: internal factors and beliefs, vaccination related factors, external influences and Covid-19 and changing perceptions of the pandemic. DISCUSSION: Findings of this study increase awareness of some of the factors influencing vaccination decisions of pregnant women. It informs practice regarding healthcare professionals' discussions with pregnant women about vaccinations, and future vaccination campaigns and interventions that are targeting an increase in vaccination uptake amongst this population.

Augmentation Therapy for Severe Alpha-1 Antitrypsin Deficiency Improves Survival and Is Decoupled from Spirometric Decline-A Multinational Registry Analysis.

Rationale: Intravenous plasma-purified alpha-1 antitrypsin (IV-AAT) has been used as therapy for alpha-1 antitrypsin deficiency (AATD) since 1987. Previous trials (RAPID and RAPID-OLE) demonstrated efficacy in preserving computed tomography of lung density but no effect on FEV1. This observational study evaluated 615 people with severe AATD from three countries with socialized health care (Ireland, Switzerland, and Austria), where access to standard medical care was equal but access to IV-AAT was not. Objectives: To assess the real-world longitudinal effects of IV-AAT. Methods: Pulmonary function and mortality data were utilized to perform longitudinal analyses on registry participants with severe AATD. Measurements and Main Results: IV-AAT confers a survival benefit in severe AATD (P < 0.001). We uncovered two distinct AATD phenotypes based on an initial respiratory diagnosis: lung index and non-lung index. Lung indexes demonstrated a more rapid FEV1 decline between the ages of 20 and 50 and subsequently entered a plateau phase of minimal decline from 50 onward. Consequentially, IV-AAT had no effect on FEV1 decline, except in patients with a Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage 2 lung index. Conclusions: This real-world study demonstrates a survival advantage from IV-AAT. This improved survival is largely decoupled from FEV1 decline. The observation that patients with severe AATD fall into two major phenotypes has implications for clinical trial design where FEV1 is a primary endpoint. Recruits into trials are typically older lung indexes entering the plateau phase and, therefore, unlikely to show spirometric benefits. IV-AAT attenuates spirometric decline in lung indexes in GOLD stage 2, a spirometric group commonly outside current IV-AAT commencement recommendations.

Shared learning to improve the care for young people and mental health within nurse education (SHYNE). Improving attitudes, confidence and self-efficacy.

Preparing students for the complexities of practice is an ongoing challenge of pre-registration nurse education. One such complexity is the increase in children and young people with mental health problems. Pre-registration student nurses from child and mental health specialisms from one University participated in an innovative simulation session, whereby actors from a youth theatre group simulated young people admitted to an acute non-mental health hospital setting for treatment of self-harm injuries. This study used an uncontrolled pre and post design to determine the impact of the session on student nurses' attitudes, confidence and self-efficacy when caring for young people who self-harm. Attitudes towards self-harm was measured using a 13 item self-report questionnaire. Confidence was measured through Likert scale responses. Self-efficacy for working with children and young people who have self-harmed was measured through an adapted version of the Self-Efficacy Towards Helping (SETH) scale. In total 101 student nurses took part in the study and 99% completed post simulation outcome measures. At post-session, the students reported a statistically significant improvement in attitudes, self-efficacy and confidence towards children and young people who self-harm. A lack of confidence is frequently reported in the literature when caring for this client group in practice settings. Improvements in attitudes, confidence and self-efficacy can positively impact individual nursing practice. Furthermore the simulation literature indicates that the skills consolidated using such an educational approach are taken forward into clinical practice. While broad claims of success should be avoided, it is promising to find a learning method that is effective in addressing a contemporary and complex health issue. Shared Learning to Improve the Care for Young People and Mental Health within Nurse Education (SHYNE): Improving Attitudes, Confidence and Self-Efficacy.

Statin use and risk of acute diverticulitis: A population-based case-control study.

The goal of the study was to examine the association between statin use and the development of acute diverticulitis requiring hospital admission.Acute diverticulitis is a common and costly gastrointestinal disorder. Although the incidence is increasing its pathophysiology and modifiable risk factors are incompletely understood. Statins affect the inflammatory response and represent a potential risk reducing agent.A retrospective, population-based, case-control study was carried out on a cohort of adults, resident in Canterbury, New Zealand. All identified cases were admitted to hospital and had computed tomography confirmed diverticulitis. The positive control group comprised patients on non-aspirin nonsteroidal anti-inflammatory drugs (NSAIDs), and the negative control group were patients on selective serotonin reuptake inhibitors (SSRIs). Medicine exposure was obtained from the Pharmaceutical Management Agency of New Zealand. Subgroup analysis was done by age and for complicated and recurrent diverticulitis.During the study period, there were 381,792 adults resident in Canterbury. The annual incidence of diverticulitis requiring hospital presentation was 18.6 per 100,000 per year. Complicated disease was seen in 37.4% (158) of patients, and 14.7% (62) had recurrent disease. Statins were not found to affect the risk of developing acute diverticulitis, nor the risk of complicated or recurrent diverticulitis. Subgroup analysis suggested statin use was associated with a decreased risk of acute diverticulitis in the elderly (age >64 years). NSAIDs were associated with a decreased risk of acute diverticulitis (risk ratio = 0.65, confidence interval: 0.26-0.46, P < .01), as were SSRIs (risk ratio = 0.37, confidence interval: 0.26-0.54, P < .01).This population-based study does not support the hypothesis that statins have a preventative effect on the development of diverticulitis, including complicated disease. We also found a decreased risk of diverticulitis associated with NSAID and SSRI use.

Ursodeoxycholic acid enriches intestinal bile salt hydrolase-expressing Bacteroidetes in cholestatic pregnancy.

Ursodeoxycholic acid (UDCA) treatment can reduce itch and lower endogenous serum bile acids in intrahepatic cholestasis of pregnancy (ICP). We sought to determine how it could influence the gut environment in ICP to alter enterohepatic signalling. The gut microbiota and bile acid content were determined in faeces from 35 pregnant women (14 with uncomplicated pregnancies and 21 with ICP, 17 receiving UDCA). Faecal bile salt hydrolase activity was measured using a precipitation assay. Serum fibroblast growth factor 19 (FGF19) and 7α-hydroxy-4-cholesten-3-one (C4) concentrations were measured following a standardised diet for 21 hours. Women with a high ratio of Bacteroidetes to Firmicutes were more likely to be treated with UDCA (Fisher's exact test p = 0.0178) than those with a lower ratio. Bile salt hydrolase activity was reduced in women with low Bacteroidetes:Firmicutes. Women taking UDCA had higher faecal lithocholic acid (p < 0.0001), with more unconjugated bile acids than women with untreated ICP or uncomplicated pregnancy. UDCA-treatment increased serum FGF19, and reduced C4 (reflecting lower bile acid synthesis). During ICP, UDCA treatment can be associated with enrichment of the gut microbiota with Bacteroidetes. These demonstrate high bile salt hydrolase activity, which deconjugates bile acids enabling secondary modification to FXR agonists, enhancing enterohepatic feedback via FGF19.

Clarifying the Risk of Lung Disease in SZ Alpha-1 Antitrypsin Deficiency.

Rationale: The ZZ genotype of alpha-1 antitrypsin deficiency (AATD) is associated with chronic obstructive pulmonary disease (COPD), even among never-smokers. The SZ genotype is also considered severe; yet, its effect on lung health remains unclear.Objectives: To determine the effect of SZ-AATD on spirometry compared with a normal-risk population and to determine the effect of smoking cessation in this genotype.Methods: We prospectively enrolled 166 related individuals, removing lung index cases to reduce bias, and compared spirometry between 70 SZ and 46 MM/MS individuals (control subjects). The effect of AAT concentrations on outcomes was assessed in 82 SZ individuals (including lung index cases). Subsequently, we analyzed retrospective SZ registry data to determine the effect of smoking cessation on spirometry decline (n = 60) and plasma anti-neutrophil elastase capacity (n = 20).Measurements and Main Results: No difference between SZ and control never-smokers was seen. Ever smoking was associated with a lower FEV1% predicted (-14.3%; P = 0.0092) and a lower FEV1/FVC ratio (-0.075; P = 0.0041) in SZ-AATD. No association was found between AAT concentration and outcomes for SZ-AATD. Longitudinal analysis of 60 SZ individuals demonstrated that COPD at baseline, but not former smoking or AAT concentrations, predicted greater spirometry decline. Finally, anti-neutrophil elastase capacity did not differ between former smokers and never-smokers (P = 0.67).Conclusions: SZ never-smokers demonstrated no increased risk of COPD, regardless of AAT concentration. Smoking interacts with SZ-AATD to significantly increase airflow obstruction. Former smoking alone is not associated with greater spirometry decline in SZ-AATD, suggesting that cessation attenuates the obstructive process. We found no evidence that the putative protective threshold or AAT concentrations predict risk within the SZ genotype, raising further doubts over the need for intravenous AAT augmentation in this cohort.

Enhanced Microbial Bile Acid Deconjugation and Impaired Ileal Uptake in Pregnancy Repress Intestinal Regulation of Bile Acid Synthesis.

Pregnancy is associated with progressive hypercholanemia, hypercholesterolemia, and hypertriglyceridemia, which can result in metabolic disease in susceptible women. Gut signals modify hepatic homeostatic pathways, linking intestinal content to metabolic activity. We sought to identify whether enteric endocrine signals contribute to raised serum bile acids observed in human and murine pregnancies, by measuring fibroblast growth factor (FGF) 19/15 protein and mRNA levels, and 7α-hydroxy-4-cholesten-3-one. Terminal ileal farnesoid X receptor (FXR)-mediated gene expression and apical sodium bile acid transporter (ASBT) protein concentration were measured by qPCR and western blotting. Shotgun whole-genome sequencing and ultra-performance liquid chromatography tandem mass spectrometry were used to determine the cecal microbiome and metabonome. Targeted and untargeted pathway analyses were performed to predict the systemic effects of the altered metagenome and metabolite profiles. Dietary CA supplementation was used to determine whether the observed alterations could be overcome by intestinal bile acids functioning as FXR agonists. Human and murine pregnancy were associated with reduced intestinal FXR signaling, with lower FGF19/15 and resultant increased hepatic bile acid synthesis. Terminal ileal ASBT protein was reduced in murine pregnancy. Cecal bile acid conjugation was reduced in pregnancy because of elevated bile salt hydrolase-producing Bacteroidetes. CA supplementation induced intestinal FXR signaling, which was not abrogated by pregnancy, with strikingly similar changes to the microbiota and metabonome as identified in pregnancy. Conclusion: The altered intestinal microbiota of pregnancy enhance bile acid deconjugation, reducing ileal bile acid uptake and lowering FXR induction in enterocytes. This exacerbates the effects mediated by reduced bile acid uptake transporters in pregnancy. Thus, in pregnant women and mice, there is reduced FGF19/15-mediated hepatic repression of hepatic bile acid synthesis, resulting in hypercholanemia.

Time to colonoscopy for patients accessing the direct access colonoscopy service compared to the normal service in Newcastle, Australia.

BACKGROUND: The 2017 National Bowel Cancer Screening Program report records a median time from positive faecal occult blood test to colonoscopy of 53 days. There is some intrinsic delay in accessing specialist medical opinion prior to colonoscopy. AIM: To examine the effect of the introduction of a Direct Access Colonoscopy Service (DACS). METHODS: Using prospectively maintained databases, patients undergoing normal service (NS) colonoscopy and those referred to DACS were compared. The primary outcome measure was the time from general practitioner (GP) referral to colonoscopy. Secondary outcome measures included the proportion of patients who met the current recommended 30 days from GP referral to colonoscopy, and the proportion of patients who waited longer than 90 days. RESULTS: There were 289 patients in the NS group, and 601 patients who progressed on the DACS pathway. The demographics of both groups were comparable. DACS patients had a median waiting time of 49 days, significantly shorter than NS patients whose median wait was 79 days (P < 0.0001). Approximately 15.1% patients in the DACS group had their colonoscopy within 30 days from GP referral, significantly better than in the NS group (4.5%, P < 0.001). In the NS group, 41.2% patients waited longer than 90 days from GP referral to colonoscopy, compared with 16.3% in the DACS group (P < 0.001). CONCLUSION: DACS reduces waiting times to colonoscopy and is associated with an increased proportion of patients undergoing colonoscopy in a timely manner.

Direct access colonoscopy service for bowel cancer screening produces a positive financial benefit for patients and local health districts.

BACKGROUND: A direct access colonoscopy service (DACS) for the National Bowel Cancer Screening Program has become standard of care in Newcastle public hospitals because of the effect it has on time to colonoscopy. Cost-effectiveness has not been studied to date. AIM: The aim of this retrospective study was to analyse the cost-effectiveness of a DACS. METHODS: Data were collected for patients referred to DACS between January 2014 and June 2016, and patients who were treated on the normal service pathway in 2013 prior to the introduction of the process. A cost-benefit analysis from the patient's and local health district's perspective was undertaken. RESULTS: Introduction of the DACS produces a direct financial gain to patients in the form of reduced direct costs. It produces an indirect financial gain in terms of increased productivity if the patient is in work, and of increased leisure time if not in work. The DACS is modest income generating for the local health district, an evaluation which is sensitive to internal policies for distribution of government funding within a district. The DACS increases the availability of outpatient consultations to other patients, which is not a quantifiable economic benefit, but is likely to be an overall health benefit. CONCLUSION: The introduction of DACS in the public system in Australia is of financial benefit to patients and to the local health service provider. It is likely to produce health benefits to non-screening patients, by means of freeing consultations to be used for other indications.

The variation in hectolitre weight of wheat grain fed with or without enzyme supplementation influences nutrient digestibility and subsequently affects performance in pigs.

A 2 × 2 factorial experiment was conducted to investigate the effect of feeding a wheat-based diet of two different hectolitre weights (66 vs. 74 kg/hl), achieved through different agronomical conditions, with or without the supplementation of a ß-glucanase and ß-xylanase enzyme mix on young pigs. The parameter categories which were assessed included growth performance, coefficient of apparent total tract digestibility (CATTD), faecal consistency, faecal microbial populations and faecal volatile fatty acid (VFA) concentrations. Sixty-four pigs (11.6 kg SD 0.97) were assigned to one of four dietary treatments: (T1) low hectolitre weight wheat diet, (T2) low hectolitre weight wheat diet containing 0.1 g/kg ß-glucanase and ß-xylanase enzyme supplement, (T3) high hectolitre weight wheat diet and (T4) high hectolitre weight wheat diet containing 0.1 g/kg ß-glucanase and ß-xylanase enzyme supplement. The inclusion of wheat was 500 g/kg in the diet. The low hectolitre weight grain had a higher level of zearalenone, aflatoxin and ochratoxin contamination compared to the high hectolitre weight grain. The high hectolitre weight wheat had a higher gross energy (GE), crude protein (CP) and lysine contents compared to the low hectolitre weight wheat. Pigs offered the low hectolitre weight diet had a lower average daily gain (ADG) (p < 0.001), a lower gain to feed (G:F) ratio (p < 0.001) and a higher faecal score (more diarrhoea) (p < 0.001) compared to pigs offered the high hectolitre weight. The low hectolitre weight diet had a reduced CATTD (p < 0.05) of nitrogen (N) and gross energy (GE) compared with pigs offered the high hectolitre weight diet. In conclusion, the higher level of mycotoxins and lower content of GE, CP and lysine in the low-quality wheat reduced ADG and the CATTD of nutrients in pigs offered this diet. The inclusion of a ß-glucanase and ß-xylanase enzyme mix had no effect on growth performance or nutrient digestibility.

A Review of the Role of Carcinoembryonic Antigen in Clinical Practice.

Carcinoembryonic antigen (CEA) is not normally produced in significant quantities after birth but is elevated in colorectal cancer. The aim of this review was to define the current role of CEA and how best to investigate patients with elevated CEA levels. A systematic review of CEA was performed, in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Studies were identified from PubMed, Cochrane library, and controlled trials registers. We identified 2,712 papers of which 34 were relevant. Analysis of these papers found higher preoperative CEA levels were associated with advanced or metastatic disease and thus poorer prognosis. Postoperatively, failure of CEA to return to normal was found to be indicative of residual or recurrent disease. However, measurement of CEA levels alone was not sufficient to improve survival rates. Two algorithms are proposed to guide investigation of patients with elevated CEA: one for patients with elevated CEA after CRC resection, and another for patients with de novo elevated CEA. CEA measurement has an important role in the investigation, management and follow-up of patients with colorectal cancer.

Therapeutic plasma exchange as a novel treatment for severe intrahepatic cholestasis of pregnancy: Case series and mechanism of action.

INTRODUCTION: Intrahepatic cholestasis of pregnancy is characterised by pruritus and elevated serum bile acids. The pruritus can be severe, and pharmacological options achieve inconsistent symptomatic improvement. Raised bile acids are linearly associated with adverse fetal outcomes, with existing management of limited benefit. We hypothesised that therapeutic plasma exchange removes pruritogens and lowers total bile acid concentrations, and improves symptoms and biochemical abnormalities in severe cases that have not responded to other treatments. METHODS: Four women with severe pruritus and hypercholanemia were managed with therapeutic plasma exchange. Serial blood biochemistry and visual analogue scores of itch severity were obtained. Blood and waste plasma samples were collected before and after exchange; individual bile acids and sulfated progesterone metabolites were measured with HPLC-MS, autotaxin activity and cytokine profiles with enzymatic methods. Results were analysed using segmental linear regression to describe longitudinal trends, and ratio t tests. RESULTS: Total bile acids and visual analogue itch scores demonstrated trends to transiently improve following plasma exchange, with temporary symptomatic benefit reported. Individual bile acids (excluding the drug ursodeoxycholic acid), and the sulfated metabolites of progesterone reduced following exchange (P = .03 and P = .04, respectively), whilst analysis of waste plasma demonstrated removal of autotaxin and cytokines. CONCLUSIONS: Therapeutic plasma exchange can lower potentially harmful bile acids and improve itch, likely secondary to the demonstrated removal of pruritogens. However, the limited current experience and potential complications, along with minimal sustained symptomatic benefit, restrict its current use to women with the most severe disease for whom other treatment options have been exhausted.

The effect of 25-hydroxyvitamin D3 and phytase inclusion on pig performance, bone parameters and pork quality in finisher pigs.

The objective of this study was to investigate the effects of supplementing both phytase and 25-hydroxyvitamin D3 (25-OH-D3) on pig performance, nutrient digestibility, carcass characteristics, bone parameters and pork quality in finisher pigs. The experimental design was a 2 × 2 factorial comprising of four dietary treatments. One hundred and twenty pigs (60 male, 60 female) were blocked according to live weight and sex and allocated to the following dietary treatments: low P (4.81 g/kg) diet (basal) (T1); low P diet + phytase (T2); low P diet + 25-OH-D3 (T3) and low P diet + phytase + 25-OH-D3 (T4). Pigs supplemented with phytase had a lower average daily feed intake (ADFI) (2.45 kg vs. 2.59 kg; p < 0.05) and lower feed conversion ratio (FCR) (2.74 kg/kg vs. 2.85 kg/kg; p < 0.05) compared to pigs offered the nonphytase diets. Pigs offered phytase diets had a higher (p < 0.05) coefficient of apparent total tract digestibility (CATTD) of ash, phosphorous (P) and calcium (Ca) compared with pigs offered the nonphytase supplemented diets. Pigs offered the 25-OH-D3 diets had a higher CATTD of N and ash. Pigs offered the phytase diets had increased (p < 0.05) bone DM, ash, Ca, P and density compared to the nonphytase diets. There was a significant interaction (p < 0.05) between phytase and 25-OH-D3 on cook loss. Pigs offered 25-OH-D3 had increased cook loss over the basal diet; however, there was no effect on cook loss when phytase and 25-OH-D3 were offered in combination compared to the phytase only diet. Pigs offered 25-OH-D3 exhibited higher (p < 0.05) Warner Bratzler shear force values and lower (p < 0.05) pork lightness (L*) surface colorimeter values. In conclusion, there was no benefit to offering a combination of phytase and 25-OH-D3 on pig performance, bone parameters or pork quality.

The use of synthetic and natural vitamin D sources in pig diets to improve meat quality and vitamin D content.

This study investigated the effects of synthetic and natural sources of vitamin D biofortification in pig diets on pork vitamin D activity and pork quality. One hundred and twenty pigs (60 male, 60 female) were assigned to one of four dietary treatments for a 55 d feeding period. The dietary treatments were (1)50 µg vitamin D3/kg of feed; (2)50 µg of 25-hydroxvitamin D3/kg of feed (25-OH-D3); (3)50 µg vitamin D2/kg of feed; (4)50 µg vitamin D2-enriched mushrooms/kg of feed (Mushroom D2). The pigs offered the 25-OH-D3 diet exhibited the highest (P < 0.001) serum total 25-hydroxyvitamin D concentration and subsequently exhibited the highest (P < 0.05) Longissimus thoracis (LT) total vitamin D activity. Mushroom D2 and 25-OH-D3 supplementation increased pork antioxidant status. The vitamin D2-enriched mushrooms improved (P < 0.05) pig performance, carcass weight and LT colour. In conclusion, 25-OH-D3 is the most successful source for increasing pork vitamin D activity, while Mushroom D2 may be a new avenue to improve animal performance and pork quality.

Vitamin D-biofortified beef: A comparison of cholecalciferol with synthetic versus UVB-mushroom-derived ergosterol as feed source.

This study investigates dietary fortification of heifer feeds with cholecalciferol and ergocalciferol sources and effects on beef total vitamin D activity, vitamer, respective 25-hydroxymetabolite contents, and meat quality. Thirty heifers were allocated to one of three dietary treatments [(1) basal diet + 4000 IU of vitamin D3 (Vit D3); (2) basal diet + 4000 IU of vitamin D2 (Vit D2); and (3) basal diet + 4000 IU of vitamin D2-enriched mushrooms (Mushroom D2)] for a 30 day pre-slaughter period. Supplementation of heifer diets with Vit D3 yielded higher (p < 0.001) Longissimus thoracis (LT) total vitamin D activity (by 38-56%; p < 0.05) and serum 25-OH-D concentration (by 20-36%; p < 0.05), compared to that from Vit D2 and Mushroom D2 supplemented animals. Irrespective of vitamin D source, carcass characteristics, sensory and meat quality parameter were unaffected (p > 0.05) by the dietary treatments. In conclusion, vitamin D3 biofortification of cattle diets is the most efficacious way to enhance total beef vitamin D activity.

Cholecalciferol supplementation of heifer diets increases beef vitamin D concentration and improves beef tenderness.

This study investigated the effects of cholecalciferol (vitamin D3) supplementation on beef vitamin D activity, beef tenderness and sensory attributes. Thirty heifers were randomly allocated to one of three finishing dietary treatments [(T1) basal diet+0IU vitamin D3; (T2) basal diet+2000IU vitamin D3; and (T3) basal diet+4000IU vitamin D3] for a 30day period pre-slaughter. Vitamin D3 supplementation linearly increased serum 25-hydroxyvitamin D3 (25-OH-D3) concentrations (R2=0.48), Longissimus thoracis (LT) total vitamin D activity (R2=0.78) as well as individually vitamin D3 (R2=0.84) and 25-OH-D3 (R2=0.75). The highest vitamin D3 inclusion diet (T3) had a 42% increase (P<0.001) in LT vitamin D activity compared to the intermediate diet (T2) and a 145% increase over the lowest level diet (T1). Vitamin D3 supplementation decreased LT shear (P<0.05) force values after 14days chilling. Sensory parameters were not affected (P>0.05). In conclusion, through short-term vitamin D3 supplementation of cattle diets, beef vitamin D activity can successfully be enhanced.

Harnessing the Potential of Mesenchymal Stem Cells for IVD Regeneration.

Intervertebral disc (IVD) degeneration is one of the leading causes of low back pain, which affects a large proportion of the global population at a huge socioeconomic burden. Current treatments focus primarily on symptomatic pain relief or surgery, but offer relatively poor long-term efficacy as they fail to address the pathogenesis of the underlying IVD degeneration. In order to offer improved clinical outcomes, a number of biological and regenerative therapies are currently being developed which target the disease at a molecular and cellular level and aim to restore IVD function. This review focusses on the considerations for development of cell-based therapies for IVD regeneration. In particular it focusses on the identification of novel progenitor cell populations within the IVD and the application of mesenchymal stem cells (MSCs) in IVD tissue engineering, which are being increasingly studied as they offer huge potential for tissue regeneration. Additionally it highlights how the growing understanding of the molecular phenotype of IVD cells is allowing tailored differentiation strategies to be developed and how MSC source and choice of growth factor influences cell phenotype and appropriate tissue formation. Finally, it reviews the range of functional biomaterials being developed to aid MSC delivery and differentiation, and discusses the potential impact the degenerate IVD microenvironmental niche may have on MSC behaviour following implantation.

Growth differentiation factor 6 and transforming growth factor-beta differentially mediate mesenchymal stem cell differentiation, composition, and micromechanical properties of nucleus pulposus constructs.

INTRODUCTION: Currently, there is huge research focus on the development of novel cell-based regeneration and tissue-engineering therapies for the treatment of intervertebral disc degeneration and the associated back pain. Both bone marrow-derived (BM) mesenchymal stem cells (MSCs) and adipose-derived MSCs (AD-MSCs) are proposed as suitable cells for such therapies. However, currently no consensus exists as to the optimum growth factor needed to drive differentiation to a nucleus pulposus (NP)-like phenotype. The aim of this study was to investigate the effect of growth differentiation factor-6 (GDF6), compared with other transforming growth factor (TGF) superfamily members, on discogenic differentiation of MSCs, the matrix composition, and micromechanics of engineered NP tissue constructs. METHODS: Patient-matched human AD-MSCs and BM-MSCs were seeded into type I collagen hydrogels and cultured in differentiating media supplemented with TGF-ß3, GDF5, or GDF6. After 14 days, quantitative polymerase chain reaction analysis of chondrogenic and novel NP marker genes and sulfated glycosaminoglycan (sGAG) content of the construct and media components were measured. Additionally, construct micromechanics were analyzed by using scanning acoustic microscopy (SAM). RESULTS: GDF6 stimulation of BM-MSCs and AD-MSCs resulted in a significant increase in expression of novel NP marker genes, a higher aggrecan-to-type II collagen gene expression ratio, and higher sGAG production compared with TGF-ß or GDF5 stimulation. These effects were greater in AD-MSCs than in BM-MSCs. Furthermore, the acoustic-wave speed measured by using SAM, and therefore tissue stiffness, was lowest in GDF6-stiumlated AD-MSC constructs. CONCLUSIONS: The data suggest that GDF6 stimulation of AD-MSCs induces differentiation to an NP-like phenotype and results in a more proteoglycan-rich matrix. Micromechanical analysis shows that the GDF6-treated AD-MSCs have a less-stiff matrix composition, suggesting that the growth factor is inducing a matrix that is more akin to the native NP-like tissue. Thus, this cell and growth-factor combination may be the ideal choice for cell-based intervertebral disc (IVD)-regeneration therapies.

A very early-branching Staphylococcus aureus lineage lacking the carotenoid pigment staphyloxanthin.

Here we discuss the evolution of the northern Australian Staphylococcus aureus isolate MSHR1132 genome. MSHR1132 belongs to the divergent clonal complex 75 lineage. The average nucleotide divergence between orthologous genes in MSHR1132 and typical S. aureus is approximately sevenfold greater than the maximum divergence observed in this species to date. MSHR1132 has a small accessory genome, which includes the well-characterized genomic islands, νSAα and νSaß, suggesting that these elements were acquired well before the expansion of the typical S. aureus population. Other mobile elements show mosaic structure (the prophage ϕSa3) or evidence of recent acquisition from a typical S. aureus lineage (SCCmec, ICE6013 and plasmid pMSHR1132). There are two differences in gene repertoire compared with typical S. aureus that may be significant clues as to the genetic basis underlying the successful emergence of S. aureus as a pathogen. First, MSHR1132 lacks the genes for production of staphyloxanthin, the carotenoid pigment that confers upon S. aureus its characteristic golden color and protects against oxidative stress. The lack of pigment was demonstrated in 126 of 126 CC75 isolates. Second, a mobile clustered regularly interspaced short palindromic repeat (CRISPR) element is inserted into orfX of MSHR1132. Although common in other staphylococcal species, these elements are very rare within S. aureus and may impact accessory genome acquisition. The CRISPR spacer sequences reveal a history of attempted invasion by known S. aureus mobile elements. There is a case for the creation of a new taxon to accommodate this and related isolates.

Evidence for a lineage of virulent bacteriophages that target Campylobacter.

BACKGROUND: Our understanding of the dynamics of genome stability versus gene flux within bacteriophage lineages is limited. Recently, there has been a renewed interest in the use of bacteriophages as 'therapeutic' agents; a prerequisite for their use in such therapies is a thorough understanding of their genetic complement, genome stability and their ecology to avoid the dissemination or mobilisation of phage or bacterial virulence and toxin genes. Campylobacter, a food-borne pathogen, is one of the organisms for which the use of bacteriophage is being considered to reduce human exposure to this organism. RESULTS: Sequencing and genome analysis was performed for two Campylobacter bacteriophages. The genomes were extremely similar at the nucleotide level (> or = 96%) with most differences accounted for by novel insertion sequences, DNA methylases and an approximately 10 kb contiguous region of metabolic genes that were dissimilar at the sequence level but similar in gene function between the two phages. Both bacteriophages contained a large number of radical S-adenosylmethionine (SAM) genes, presumably involved in boosting host metabolism during infection, as well as evidence that many genes had been acquired from a wide range of bacterial species. Further bacteriophages, from the UK Campylobacter typing set, were screened for the presence of bacteriophage structural genes, DNA methylases, mobile genetic elements and regulatory genes identified from the genome sequences. The results indicate that many of these bacteriophages are related, with 10 out of 15 showing some relationship to the sequenced genomes. CONCLUSIONS: Two large virulent Campylobacter bacteriophages were found to show very high levels of sequence conservation despite separation in time and place of isolation. The bacteriophages show adaptations to their host and possess genes that may enhance Campylobacter metabolism, potentially advantaging both the bacteriophage and its host. Genetic conservation has been shown to extend to other Campylobacter bacteriophages, forming a highly conserved lineage of bacteriophages that predate upon campylobacters and indicating that highly adapted bacteriophage genomes can be stable over prolonged periods of time.