Thermally triggered hydrogel injection into bovine intervertebral disc tissue explants induces differentiation of mesenchymal stem cells and restores mechanical function.

We previously reported a synthetic Laponite® crosslinked pNIPAM-co-DMAc (L-pNIPAM-co-DMAc) hydrogel which promotes differentiation of mesenchymal stem cells (MSCs) to nucleus pulposus (NP) cells without additional growth factors. The clinical success of this hydrogel is dependent on: integration with surrounding tissue; the capacity to restore mechanical function; as well as supporting the viability and differentiation of delivered MSCs. Bovine NP tissue explants were injected with media (control), human MSCs (hMSCs) alone, acellular L-pNIPAM-co-DMAc hydrogel or hMSCs incorporated within the L-pNIPAM-co-DMAc hydrogel and maintained at 5% O2 for 6weeks. Viability of native NP cells and delivered MSCs was maintained. Furthermore hMSCs delivered via the L-pNIPAM-co-DMAc hydrogel differentiated and produced NP matrix components: aggrecan, collagen type II and chondroitin sulphate, with integration of the hydrogel with native NP tissue. In addition L-pNIPAM-co-DMAc hydrogel injected into collagenase digested bovine discs filled micro and macro fissures, were maintained within the disc during loading and restored IVD stiffness. The mechanical support of the L-pNIPAM-co-DMAc hydrogel, to restore disc height, could provide immediate symptomatic pain relief, whilst the delivery of MSCs over time regenerates the NP extracellular matrix; thus the L-pNIPAM-co-DMAc hydrogel could provide a combined cellular and mechanical repair approach. STATEMENT OF SIGNIFICANCE: Low back pain (LBP) is associated with degeneration of the intervertebral disc (IVD). We have previously described development of a jelly delivery system (hydrogel). This has the potential to deliver adult stem cells to the centre of the IVD, known as the nucleus pulposus (NP). Here, we have demonstrated that adult stem cells can be safely injected into the NP using small bore needles, reducing damage to the disc. Following injection the hydrogel integrates with surrounding NP tissue, promotes differentiation of stem cells towards disc cells and restores IVD mechanical function. The hydrogel could be used to restore mechanical function to the IVD and deliver cells to promote regeneration of the disc as a minimally invasive treatment for LBP.

Short communication: Rumination and feeding behavior before and after calving in dairy cows.

The objectives of the current study were to describe changes in rumination and feeding behavior around calving. Rumination time, feeding time, and dry matter intake were monitored in 11 freestall-housed cows from 96 h before to 48 h after calving. Data were summarized in 2-h and 24-h periods, adjusting for calving time. Differences between baseline (96 to 24h before calving) and subsequent 24-h periods were evaluated. Compared with baseline, cows spent, on average, 63 ± 30 min/24h less time ruminating and 66 ± 16 min/24h less time feeding in the 24-h period before calving. These behaviors continued to decline during the 24-h period after calving when, compared with baseline, time spent ruminating decreased on average by 133 ± 35 min/24h and time spent feeding decreased by 82 ± 18 min/24h. Dry matter intake tended to decrease by 3.8 ± 1.9 kg in the 24-h period before calving but returned to baseline values in the 24-h following calving. Rumination time and time spent feeding started to decline approximately 4 and 8h before calving, respectively, and increased in the 4 to 6h following calving. Rumination time and time spent feeding show promise as tools to identify cows close to calving.

Feeding a higher forage diet prepartum decreases incidences of subclinical ketosis in transition dairy cows.

A common feeding practice during the dry period is to switch dairy cows to an energy-dense diet 3 wk prepartum, but this practice may lead to the overconsumption of energy and increase the risk of metabolic disease postpartum. The aim of this trial was to compare the metabolic status of transition Holstein dairy cows fed a 77% forage diet (77F; NEl = 1.46 Mcal/kg; NDF = 41%) vs. those fed an 87% forage diet (87F; NEl = 1.41 Mcal/kg; 48% NDF). Approximately 60 d before calving, cows were dried off, housed in a free stall barn, and fed the 87F diet. Three weeks before expected calving, cows were randomly assigned to either the 77F treatment and switched to this diet (n = 45) or assigned to the 87F treatment and stayed on the dry cow ration until parturition (n = 42). After parturition, all cows were fed a common lactation diet (NEl = 1.59 Mcal/kg; 36% NDF). Dry matter intake was measured daily from 2 wk before to 2 wk after calving. Blood was sampled daily for 10 d postpartum. Subclinical ketosis was diagnosed using a threshold of ß-hydroxybutyrate (BHBA) ≥ 1.0 mmol/L after calving. The percentage of cows pregnant and odds of being pregnant for each treatment group were determined at 60, 90, and 120 d in milk through ultrasound by the herd veterinarian. Cows on the 87F diet consumed less DM prepartum than those on the 77F diet (12.7 ± 0.3 kg/d vs. 15.4 ± 0.3 kg/d, P < 0.001), but no difference in DMI was detected after calving (19.7 ± 5.5 kg/d; P = 0.87). Although the calculated prepartum required energy intake was the same for the 2 treatments (15.3 ± 1.2 Mcal/d; P = 0.16), cows on the 77F diet consumed 4.5 Mcal/d more than those on the 87F diet (22.5 ± 0.5 Mcal/d vs. 18.0 ± 0.5 Mcal/d; P < 0.001). Postpartum concentration of BHBA was less for cows fed the 87F diet prepartum (0.49 ± 0.02 mmol/L vs. 0.59 ± 0.02 mmol/L; P = 0.02), and fewer animals on this diet were diagnosed subclinical ketosis (SCK; 49% vs. 17%; P = 0.001). Milk production tended to be less for cows fed the 87F diet prepartum (47.3 ± 0.4 kg/d vs. 48.8 ± 0.4 kg/d; P = 0.10) for the first 22 wk of lactation, which was significant for d 7 to 28 of lactation (44.6 ± 1.1 kg/d vs. 47.6 ± 1.0 kg/d; P = 0.05). Although sample size was small to draw strong conclusions on reproductive performance, at 120 d in milk, cows on the 87F diet were 0.3 times more likely to be pregnant (P = 0.03). These results indicate that feeding an 87F diet before calving can reduce rates of SCK in transition dairy cows.

Technical note: Comparison of rectal and vaginal temperatures in lactating dairy cows.

A method commonly used to identify illness in dairy cows is measuring body temperatures with a rectal thermometer, but vaginal measures are becoming common in research. The primary objective of this study was to validate vaginal measures of body temperature by comparing them with rectal temperatures. Data loggers used to collect vaginal temperatures can be programmed to collect many readings per day, providing an opportunity to interpret effects of health in relation to diurnal differences in temperatures. Thus, a secondary objective was to compare the diurnal pattern in body temperatures for cows with and without retained placenta (RP). Body temperature was monitored for 8 d in 29 cows that had recently calved (enrolled 2 d after calving; 7 of these cows were diagnosed with RP) and in 13 cows in peak lactation (98±8 d in milk). Rectal temperatures were taken at 0630, 0930, 1230, 1530, 1830, and 2130h (±30 min) with a digital thermometer for 8 d consecutively. During the same period, vaginal temperatures were measured every 10 min with a microprocessor-controlled data logger attached to a modified vaginal controlled internal drug release insert. Values from the vaginal loggers were averaged over 1h and paired with the corresponding rectal temperature. There was a relationship between rectal and vaginal temperatures for fresh cows (n=1,393; r=0.81) and for peak-lactation cows (n=556; r=0.46). Cows with RP had higher body temperatures (39.2±0.01) compared with healthy cows (39.1±0.01). Body temperature was higher at night, and lower between 0800 to 1000 h for healthy cows (39.0±0.02) and between 1100 to 1300 h for RP cows (39.1±0.02). In summary, vaginal temperatures were associated with rectal measures, and provided the advantage of capturing dirurnal changes in body temperature.

Increased titres of anti-human heat shock protein 60 predict an adverse one year prognosis in patients with acute cardiac chest pain.

OBJECTIVE: To assess whether antibodies to human heat shock protein 60 (anti-huhsp60) or to mycobacterial heat shock protein 65 (anti-mhsp65) predict an adverse one year prognosis in patients admitted with acute cardiac chest pain. DESIGN: Prospective observational study. SETTING: Teaching hospital. PATIENTS: 588 consecutive emergency admissions of patients with acute chest pain of suspected cardiac origin. MAIN OUTCOME MEASURES: Anti-huhsp60 and anti-mhsp65 titres were assayed on samples drawn on the morning after admission. The end points after discharge were coronary heart disease death, non-fatal myocardial infarction, coronary artery bypass grafting, percutaneous transluminal coronary angioplasty, angiogram, or readmission with further cardiac ischaemic chest pain. RESULTS: During follow up after discharge (mean of 304 days, range 1-788 days), 277 patients had at least one of the study outcomes. Patients with increased titres of anti-huhsp60 had an adverse prognosis (hazard ratio 1.56 (95% confidence interval 1.09 to 2.23) comparing highest versus lowest quartiles, p = 0.015). Anti-mhsp65 titres were not predictive. CONCLUSIONS: Patients admitted with acute cardiac chest pain and increased titres of anti-huhsp60 had an adverse one year prognosis.

How evidence based are recruitment strategies to randomized controlled trials in primary care? Experience from seven studies.

BACKGROUND: Failure to recruit adequate numbers of participants represents a major barrier to the completion of randomized controlled trials in primary care and is associated with substantial opportunity costs. However, uncertainty exists regarding the relative effectiveness of different methods to promote recruitment. OBJECTIVES: The purpose of this study was to estimate the proportion of strategies used to promote patient recruitment to randomized controlled trials in primary care that are evidence based. METHODS: Investigators from seven primary care-based clinical trials of dyspepsia management aiming to recruit a total of 6070 patients participated. Following a survey of trial organization, a Delphi technique was used to reach consensus on levels of evidence on the effectiveness of interventions or organizational characteristics in influencing recruitment. The main outcome measures were the proportions of interventions or organizational characteristics for influencing patient recruitment that are based upon randomized controlled trials, on convincing non-experimental evidence or meeting neither of these criteria. RESULTS: Out of a total of 56 interventions used across the trials, 35 (63%) were judged as evidence based. Out of a total of 29 organizational characteristics possessed by the trials, five (17%) were judged as evidence based. Across the seven dyspepsia trials, the presence of 'favourable' organizational characteristics appeared to be important contributors towards successful recruitment. CONCLUSIONS: A wide range of interventions and organizational characteristics with the potential to promote recruitment were used or possessed by seven primary care trials. Many were not evidence based. Our experience suggests that organizational characteristics could be more influential in trial recruitment than the use of specific interventions. Given the costs of primary care-based trials, researchers need more rigorous evidence to inform recruitment strategies.

The quality evaluation program for plutonium pits at the U.S. DOE Pantex plant.

The United States Department of Energy Pantex Plant quality evaluation program for plutonium pits is an extensive program that includes 1) weigh and leak check system; 2) radiography; and 3) dye penetrant testing. Successful completion of these diagnostics qualifies a pit to remain in the active status stockpile program. The use of lead aprons and a robot when handling the plutonium pits minimizes personnel exposures to ionizing radiation. All personnel exposures to ionizing radiation at Pantex Plant are As Low As Reasonably Achievable.

A randomized trial of dehumidification in the control of house dust mite.

BACKGROUND: House dust mites (HDM) are sensitive to humidity. Few studies have adequately examined the potential of dehumidification in reducing HDM numbers. OBJECTIVE: The study examined the effect of portable domestic dehumidifiers, and a behavioural programme to reduce humidity, on HDM counts and HDM allergen levels. METHODS: A randomized controlled trial was undertaken. A total of 76 homes were allocated to three groups that received a portable domestic dehumidifier, a behavioural programme, or no intervention. Humidity, temperature, HDM counts (trap and vacuum samples), HDM allergen, and other details of the home environment were measured on four occasions over a period of 1 year. Interventions and measurements were concerned predominantly with one bedroom. RESULTS: There was a reduction in relative humidity in the dehumidifier group, but not the behavioural group. A decline in HDM trap counts was observed for all three groups. Change scores did not indicate that the dehumidifier group had a greater decline than the other groups. A secondary analysis examining presence or absence of HDM showed a shift from households having HDM at baseline to households not having HDM in the final round for some trap measures. Change score analysis indicated that this shift was greater in the dehumidifier group compared with other groups. The dehumidifier group did not show a greater decline in HDM allergen than that seen in the other two groups. CONCLUSION: Neither the dehumidifier nor the behavioural intervention had a major effect on HDM counts or allergen levels. However, the study did have a number of limitations relating to size, timing of intervention, and running of the dehumidifiers. The secondary data analysis may indicate some effect of dehumidification, but clearly this effect was small. There is a need for more information on the effects of reducing ambient humidity on the distribution of HDM within their habitats.

The dissolution vessel for plutonium pits at the U.S. DOE Pantex Plant.

The U.S. DOE Pantex Plant has been given the mission to recertify and requalify plutonium pits for reuse in existing War Reserve nuclear weapons. The first process common to both recertification and requalification is cleaning the plutonium pit. The pit will be cleaned in a dissolution vessel using N-methyl pyrrolidone (NMP) solvent. The recertification and requalification programs are both in the design concept phase at Pantex Plant The U.S. DOE Pantex Plant secures the national security of the United States by using safe vessels for cleaning plutonium pits in a manner that protects the health and safety of employees, the public and the environment.

The use of patient registers to estimate migration.

This article presents information discussed at the recent regional seminars on population statistics. It describes a new data source for estimating internal migration between local and health authorities in England and Wales, discusses the way that the data have been collected and processed, and how limitations in the raw data have been addressed. It then goes on to explain the way that the new internal migration estimates have been used in the calculation of population estimates for local and health authorities. This is a change to the method of estimating migration in the calculation of population estimates.

Expression of the Lewis group carbohydrate antigens during Xenopus development.

We have examined the pattern of expression of the Lewis group carbohydrate antigens during the development of African toad Xenopus laevis. One of these antigens, Lewis x (Le(x), also known as SSEA-1), was previously shown to be involved in cell-cell adhesion in early mouse embryos and teratocarcinoma stem cells. Recently another member of these antigens, sialyl-Le(x), was found to be one of the major ligands for the selectin family of cell-cell adhesion molecules. In order to study the role of carbohydrate-mediated cell adhesion during Xenopus development, we first studied the expression pattern of the Le(x). We found that Le(x)was not expressed in early embryos, started to be expressed at the tail bud stage in anterior regions of the body such as the cement gland or head skin, and was gradually showed more posterial expression at later stages. At tadpole stage, it was also expressed on specific cell bodies in brain, and in axon region in brain and neural retina. Antibodies against Le(x)blocked neurite outgrowth in the explant culture of tadpole brain. One of the candidates for Le(x)carrier protein in the tadpole brain is a 200 kDa glycoprotein detected by Western blotting. In adult tissues, it was expressed in brain, testis, and gut, but not in kidney, lung, spleen, ovary, or muscle. We also examined the expression patterns of other Lewis group antigens. Among them, sialyl-Le(x)was expressed on endothelial cells and on leukocytes, suggesting the possibility that it functions as a ligand for selectin in Xenopus.

The AL-R8 SI: the next generation staging container for plutonium pits at the USDOE Pantex Plant.

The AL-R8 SI (sealed insert) is the next generation staging container for plutonium pits at the U.S. DOE Pantex Plant. The sealed insert is a stainless steel container that will be placed inside a modified AL-R8 container to stagepits. A pit is a hollow sphere of plutonium metal which is the primary fissionable material in nuclear weapons (warheads and bombs). It is hermetically sealed by a cladding material, which is usually stainless steel. Personnel exposures to ionizing radiation from the pits in storage are expected to decrease due to the attenuation provided by the new SI. All personnel exposures to ionizing radiation at Pantex Plant are As Low As Reasonably Achievable (ALARA). Pantex Plant secures the common defense and national security of the United States by safely staging plutonium pits in a manner that protects the health and safety of employees, the public, and the environment.

The new subnational population projections model: methodology and projection scenarios.

This article describes the most recent process of projecting population at the subnational level in England. It briefly explains the reasons why projecting population at the subnational level is important, describes the model and how it was used to produce the latest set of long-term subnational population projections in England published by the Office for National Statistics (ONS) in 1998. The article then discusses how the model may be applied to answer various 'what-if' questions about future population.

Trends in migration in the UK.

This article presents the latest annual statistics for international migration and for internal population movements in the United Kingdom, and compares them with recent trends. It also aims to give some information on the characteristics of the migrants from the data sources that are available. Measuring migration is not straightforward as there is no compulsory system within the United Kingdom to record movements of the population, either into the country from abroad or within the country. Nevertheless, we can estimate these movements from available data sources and so monitor migration trends and patterns.

An interdisciplinary home healthcare program for patients with Parkinson's disease.

The long-term treatment of Parkinson's disease presents a special challenge in the home healthcare setting. This article describes a successful interdisciplinary home healthcare program that addresses the special needs of patients with Parkinson's disease and associated disorders. The program, which combines skilled assessment, professional healthcare services, patient and family education, and case management to optimize patient outcomes, has become a national model for agencies hoping to start or expand Parkinson's disease programs.

Modified mRNA rescue of maternal CK1/8 mRNA depletion in Xenopus oocytes.

This work addresses two issues, the use of antisense oligodeoxynucleotides to deplete specific mRNAs in Xenopus oocytes to analyze their functions during development and the role of cytokeratin filaments in cells of the early Xenopus embryo. We have shown previously that depletion of cytokeratin CK1/8 mRNA causes defects in the early embryo. In this study, we show that the oligos, modified with phosphoramidate linkages to improve stability, are capable of degrading exogenous mRNA up to 27 hours after injection in the oocyte. For this reason, the phenotype could not be rescued by injection of a synthetic CK1/8 mRNA. However, modification of the synthetic CK1/8 mRNA, which prevents annealing of the antisense oligonucleotide used for depleting the endogenous CK1/8 mRNA, did result in the rescue of the CK1/8 depletion phenotype. These results demonstrate that the phenotype observed after depletion of the CK1/8 mRNA is truly caused by the lack of CK1/8 protein. Injection of the closely related type II cytokeratin (CK55) did not result in the same level of rescue of the CK1/8 depletion phenotype, suggesting that structurally similar members of the cytokeratin family, expressed at different stages of development, cannot substitute for each other in the early embryo.

Impact of a collaborative immunisation programme in an inner city practice.

OBJECTIVES: To describe the impact of a collaborative immunisation programme, between an inner city practice and the Eastern Health Board (EHB). DESIGN: An observational study using a computer database formed from practice and EHB records. SETTING: One Dublin inner city practice with three partners located in an area with a deprived socio-economic profile. SUBJECTS: All patients in the practice aged more than six months and less than five years identified both from practice registers and opportunistically during study period. RESULTS: 342 children, older than six months and less than five years were identified at start and 464 (a 36% increase) by end of the programme. Uptake changed for DPT from 30% before, to 57% after the programme (p < 0.0005), for DT from 15% to 13%, for Hib from 7% to 50% (p < 0.0005) and for MMR (over 15 months) from 53% to 75% (p < 0.0005). Uptake of the DPT, Hib and MMR was 35% among GMS eligible, 51% among GMS ineligible (p < 0.005). CONCLUSION: A collaborative immunisation programme significantly improved practice uptake rates. These improved rates still do not attain declared national targets. To achieve these targets, radical overhaul of the immunisation service is required.

The gender-specific chest wall thickness prediction equations for routine measurements of 239Pu and 241Am within the lungs using HPGe detectors.

The current chest wall thickness prediction equation is not applicable to use in routine lung counting measurements for detection of low energy photons (17--60 keV) within the lungs of male and female subjects. The current chest wall thickness prediction equation was derived for the NaI-CsI "phoswich" detection system, which is not the routine detection system in use; the subject position was supine, which is not the routine position; the equation did not account for the intercostal tissue thicknesses of muscle and adipose which significantly attenuate low energy photons (17--60keV); it was derived from male subjects only and is used to predict the chest wall thickness of female subjects for whom it is not applicable. The current chest wall thickness prediction equation yields unacceptable percent errors in the HPGe detection efficiency calibration for 239Pu and 241Am (17- and 59.5-keV photons, respectively) relative to the gender-specific HPGe chest wall thickness prediction equations of this paper (+284% to --73% for 239Pu; (+)42% to --39% for 241Am). As a result, use of the current chest wall thickness prediction equation yields unacceptable percent errors (proportional in magnitude to the percent errors in the detection efficiency calibration) in the calculation of the minimum detectable activity (Bq) or in an initial assessment of a radioactive contamination exposure detected by a routine lung count measurement.

Involvement of a neutral glycolipid in differential cell adhesion in the Xenopus blastula.

Many different molecular species mediate cell adhesion during embryonic development. These can have either protein or carbohydrate functional groups, which can act in either a homophilic or a heterophilic manner, and often in concert. We report here that a monoclonal antibody, M4B, raised against Xenopus blastomere membranes, inhibits the calcium-dependent adhesion of dissociated blastomeres. M4B maintains its inhibitory effect on adhesion when converted into univalent fragments, and specifically affects calcium-dependent adhesion. The antigen is regulated in both space and time during early development. It is found on cell surfaces throughout the egg to blastula stages, but is more concentrated on cells in the animal and marginal zones of the blastula. It is dramatically downregulated during gastrulation, and becomes largely restricted to gut epithelium by the larval stages. We show also that M4B function is spatially differentiated at the blastula stage, since it inhibits the aggregation of dissociated animal cells to a greater extent than vegetal cells. This membrane antigen may therefore play a role in the differential adhesion observed between different regions of the blastula, and which we presume to underlie the segregation of the primary germ layers during gastrulation. M4B recognizes a complex of plasma membrane glycolipids. Periodate treatment destroys the ability of these glycolipids to react with the antibody, indicating that the epitope resides in the carbohydrate moiety of the glycolipids. Chemical characterization shows that it is a neutral glycolipid, and that the major component is of the glycoglycerolipid, rather than the more common glycosphingolipid class. Blocking experiments with oligosaccharides of defined structure, and antibody crossreactivity show that the M4B antibody does not recognize several known embryonic carbohydrate antigens. These results demonstrate that M4B antibody recognizes a novel group of developmentally regulated glycolipids which function in calcium-dependent cell--cell adhesion in the Xenopus blastula.