Applicability and safety of discontinuous ADVanced Organ Support (ADVOS) in the treatment of patients with acute-on-chronic liver failure (ACLF) outside of intensive care.

BACKGROUND: ADVanced Organ Support (ADVOS) is a novel type of extracorporeal albumin dialysis and holds promise to sustain liver function and recovery of patients with acute-on-chronic liver failure (ACLF). Previously, ADVOS was tested as continuous treatment for intensive care patients with liver failure. Data related to the applicability and safety as discontinuous treatment outside of ICU is not available. AIM: Evaluation of ADVOS as discontinuous treatment for patients with ACLF outside intensive care unit and comparison with a matched historic cohort. METHODS AND RESULTS: In this retrospective study, 26 patients with ACLF and the indication for renal replacement therapy related to HRS-AKI were included. Majority of patients were male (65%) with alcoholic cirrhosis in 88% and infections as a trigger of ACLF in 96%. Liver function was severely compromised reflected by high median MELD and CLIF-C ACLF scores of 37 (IQR 32;40) and 56.5 (IQR 51;60), respectively. Patients were treated discontinuously with ADVOS over a median time of 12 days (IQR 8.25;17) and received 8 (IQR 4.25;9.75) treatment cycles on average. No treatment related adverse events were recorded, and safety laboratory parameters remained constant during the observation time. After 16 h cumulative dialysis therapy, ADVOS significantly reduced protein-bound bilirubin (14%), creatinine (11.8%) and blood urea nitrogen (BUN, 33%). Using a matched cohort with ACLF treated with hemodialysis, ADVOS achieved a stronger decrease in bilirubin (p = 0.01), while detoxification of water-soluble catabolites' including creatinine and BUN was comparable. The 28-days mortality in the ADVOS group was 56% (14/26) and was not inferior to predicted survival (predicted median 28-days mortality was 44%, IQR 30; 59). CONCLUSION: Discontinuous ADVOS treatment was safe and effective in patients with ACLF outside intensive care and outperformed hemodialysis in reducing protein-bound metabolites.

The use of host cell DNA methylation analysis in the detection and management of women with advanced cervical intraepithelial neoplasia: a review.

This paper briefly reviews the role of hypermethylation of host cell genes in cervical carcinogenesis and discusses potential clinical applications of methylation analysis in the management of high-risk HPV (hrHPV) -positive women. We argue that methylation assays can be used: 1. for primary triage of hrHPV-positive women to detect cervical cancer and advanced cervical intraepithelial neoplasia (CIN); 2. as secondary triage for women with minor cytological abnormalities to identify those with the highest risk of CIN3 or worse; 3. as exit test for women leaving the screening programme to identify cervical cancer and advanced CIN; and 4. to support management of CIN. TWEETABLE ABSTRACT: This paper discusses potential clinical applications of DNA methylation analysis in the management of women with a high-risk HPV infection.

Associations between farmer participation in veterinary herd health management programs and farm performance.

In the past few decades, farms have increased in size and the focus of management has changed from curative to preventive. To help farmers cope with these changes, veterinarians offer veterinary herd health management (VHHM) programs, whose major objective is to support the farmer in reaching his farm performance goals. The association between farm performance and participation in VHHM, however, remains unknown. The aim of this paper was to compare farm performance parameters between participants and nonparticipants in VHHM and to differentiate within participation to evaluate the possible added value of VHHM on the farm. Five thousand farmers received a questionnaire about the level of VHHM on their farm. Farm performance parameters of these 5,000 farms were provided. For all respondents (n=1,013), farm performance was compared between participants and nonparticipants and within level of participation, using linear mixed and linear regression models. Farmers who participated in VHHM produced 336 kg of milk/cow per year more and their average milk somatic cell count (SCC) was 8,340 cells/mL lower than farmers who did not participate in VHHM. Participating herds, however, had an older age at first calving (+12d), a lower 56-d nonreturn rate percentage (-3.34%), and a higher number of inseminations per cow (+0.09 inseminations). They also had more cows culled per year (+1.05%), and a lower age at culling (-70 d). Participants in the most-extended form of VHHM (level 3) had a lower SCC (-19,800 cells/mL), fewer cows with high SCC (-1.70%), fewer cows with new high SCC (-0.47%), a shorter calving interval (-6.01 d), and fewer inseminations per heifer (-0.07 inseminations) than participants in the least-extended form of VHHM (level 1). Level 3 participants, however, also had more cows culled per year (+1.74%) and a lower age at culling (-103 d). Discussing specific topics with the veterinarian (milk production, fertility, and udder health) had only marginal effects on improving the farm performance parameters related to those topics. Given the relevance of fertility on the farm and the focus on longevity by society, it is important to determine underlying reasons for the negative associations of these topics with participation in VHHM. A longitudinal study could provide answers to this. For now, veterinarians should be aware of the associations. The increased milk production and milk quality could help the marketing of VHHM to farmers.

Veterinary herd health management programs on dairy farms in the Netherlands: use, execution, and relations to farmer characteristics.

Veterinary herd health management (VHHM) programs are of growing importance to the dairy industry; they support farmers in the shift from curative to preventive health management, caused by increased herd sizes and quality standards in dairy farming. Farmers participating in VHHM are visited every 4 to 6 wk by their veterinarian, who checks the animals and herd management to intervene in a proactive way with problems regarding animal health and animal welfare. At present, no good overview exists of how VHHM is executed on Dutch dairy farms, and whether different farmers require different types of VHHM. Aims of this study were to (1) map out how many farmers participate in VHHM, (2) describe how VHHM is executed on the farms, and (3) see whether certain farmer characteristics are related to farmers' participation in VHHM. In 2011, a questionnaire was sent to 5,000 Dutch dairy farmers per e-mail. Part 1 of the questionnaire focused on participation in and execution of VHHM and part 2 focused on farmer characteristics regarding external information. Returned questionnaires (n=1,013) were summarized and statistically analyzed. In this study 68.6% of the responding farmers participated in any form of VHHM. The most important activities were fertility checks and advice about fertility; the least important were housing and claw health. Relationships between farmer characteristics (use of and trust in information) and participation in VHHM were found.

Short communication: influence of sampling interval on the accuracy of predicting bulk milk somatic cell count.

Although bulk milk somatic cell count (BMSCC) is, in most instances, not a good proxy for actual average herd somatic cell count (SCC), BMSCC is the only SCC parameter available to monitor trends in udder health for a large number of farms worldwide. The frequency of sampling BMSCC varies considerably between countries, and it is unknown to what extent the sampling interval of BMSCC or variation in BMSCC data itself influences the accuracy. The aim of this study was to assess the effect of sampling interval and variation of the BMSCC data on the accuracy to predict BMSCC. Because BMSCC is measured at regular time intervals, an artificial neural network (ANN) was used to determine both the effect of sampling interval and variation of the BMSCC data. The intervals examined in this study ranged from 4 to 14 d and were compared with the baseline of a standard 2-d sampling interval. The BMSCC data were collected every other day for a 24-mo period on 949 farms, and all series were created by exclusion of BMSCC data in between the original 2-d sampling interval series. The effect of variation of BMSCC was determined by comparing the error of the ANN model in 2 subsets of farms, those with the lowest SD (n=239) and those with a high SD of BMSCC data (n=236). No significant differences were found in any of the sampling intervals between the 2 cohorts of low and high SD in BMSCC. Overall, compared with the 2-d sampling interval, on average the error of the ANN model was 32,600 cells/mL for all farms included, ranging from 15,000 cells/mL (4-d interval) to 41,000 cells/mL (14-d sampling interval). Therefore, the length of the sampling interval greatly influences the usefulness of BMSCC data to monitor trends in udder health at the herd level.

Prediction of the herd somatic cell count of the following month using a linear mixed effect model.

An accurate prediction of the average somatic cell count (SCC) for the next month would be a valuable tool to support udder health management decisions. A linear mixed effect (LME) model was used to predict the average herd SCC (HSCC) for the following month. The LME model included data on SCC, herd characteristics, season, and management practices determined in a previous study that quantified the contribution of each factor for the HSCC. The LME model was tested on a new data set of 101 farms and included data from 3 consecutive years. The farms were split randomly in 2 groups of 50 and 51 farms. The first group of 50 farms was used to check for systematic errors in predicting monthly HSCC. An initial model was based on older data from a different part of the Netherlands and systematically overestimated HSCC in most months. Therefore, the model was adjusted for the difference in average HSCC between the 2 sets of farms (from the previous and current study) using the data from the first group of 50 farms. Subsequently, the data from the second group of 51 farms were used to independently assess this final model. A null model (no explanatory variables included) predicted 48 and 59% of the HSCC within the predetermined range of 20,000 and 30,000 cells/mL, respectively. The final LME model predicted 72 and 81% of the HSCC of the next month correctly within these 2 ranges. These outcomes indicate that the final LME model was a valid additional tool for farmers that could be useful in their short-term decisions regarding udder health management and could be included in dairy herd health programs.

[Evaluation of veterinary advice given to dairy farmers]. / Beoordeling van de bedrijfsadvisering door dierenartsen op rundveebedrijven.

In this pilot study, the manner and effectiveness of advice given by four cattle practitioners was investigated during health management visits to 34 dairy farms. Farmers were asked about their satisfaction with the advice given, and an independent observer evaluated the manner in which practitioners carried out the farm visit. There were no major differences between the cattle practitioners, but there were minor differences in the way practitioners gave advice, in the duration of the herd visit, and in the subjects talked about. The farmers were positive about the manner in which advice was given and its effect. The method used in this pilot study could be used to gain insight into the manner in which veterinary advice is given by veterinary practitioners and veterinary practices, and may contribute to improving the quality of veterinary services provided.

Effect of herd characteristics, management practices, and season on different categories of the herd somatic cell count.

In this study, the contribution of management practices, herd characteristics, and seasonal variables to the herd somatic cell count (SCC) was quantified in herds with low (<150,000 cells/mL), medium (150,000-200,000 cells/mL), and high (>200,000 cells/mL) herd SCC (HSCC). Selection of the variables was performed using a linear mixed effect model; HSCC was calculated as the arithmetic mean of the individual cow's SCC. The data concerning management practices were derived from 3 questionnaires on mastitis prevention and management practices on 246 Dutch dairy farms. The monthly Dairy Herd Improvement test data of these 246 farms were used to calculate the herd characteristics and seasonal effects. None of the management practices were associated with HSCC in all 3 HSCC categories. Some variables only had a significant association with HSCC in one HSCC category, such as dry premilking treatment (-9,100 cells/mL in the low HSCC category) or feeding calves with high SCC milk (11,100 cells/ mL in the medium HSCC category). Others had an opposite effect on HSCC in different HSCC categories, such as average parity (-6,400 and 11,000 cells/mL in the low and medium HSCC category, respectively) and feeding calves with fresh milk (10,300 and -9,700 cells/ mL in the low and high HSCC category, respectively). We conclude that, given the individual Dairy Herd Improvement data and information on management practices of an individual farm, it is possible to provide quantitative insight into the contribution of these different variables to the HSCC of an individual farm. Being able to provide such insight is a prerequisite for interpretation, prediction, and control of HSCC on individual dairy farms.

Short communication: Comparison of bulk milk, yield-corrected, and average somatic cell counts as parameters to summarize the subclinical mastitis situation in a dairy herd.

In this study, the correlation was determined between the prevalence of high cow-level somatic cell count (SCC >250,000 cells/mL), a summary of the subclinical mastitis situation in a dairy herd, and 3 average herd SCC parameters: bulk milk SCC (BMSCC), yield-corrected test-day SCC (CHSCC), and the arithmetic average test-day SCC (HSCC) of the lactating herd. The herd prevalence of cows with an SCC of >250,000 cells/mL was calculated by using Dairy Herd Improvement data. Herds were included if BMSCC was sampled within 2 d of the Dairy Herd Improvement test day and if the BMSCC did not exceed 400,000 cells/mL. The interval between sampling, 0, 1, or 2 d, did not significantly influence the correlation between BMSCC and the prevalence of high SCC. The correlations between the prevalence of high SCC and BMSCC, yield-corrected test-day SCC, and HSCC, examined by using a linear regression model, were 0.64, 0.78, and 0.89, respectively. Therefore, it can be concluded that, based on the highest correlation, HSCC is a more appropriate parameter than BMSCC to summarize the average herd subclinical mastitis situation in a dairy herd.

Structure and function of bacterial cold shock proteins.

Cold shock proteins (Csps) comprise a family of small proteins that are structurally highly conserved and bind to single-stranded nucleic acids via their nucleic acid binding motifs RNP1 and RNP2. Bacterial Csps are mainly induced after a rapid temperature downshift to regulate the adaptation to cold stress, but are also present under normal conditions to regulate other biological functions. The structural unit characteristic for Csps occurs also as a cold shock domain (CSD) in other proteins and can be found in wide variety of organisms from bacteria to vertebrates. Important examples are the Y-box proteins that are known to be involved in regulation of several transcription and translation processes. This review describes the role of Csps in protein expression during cold shock with special emphasis on structural aspects of Csps.

Pressure-stability of phospholipid bicelles: measurement of residual dipolar couplings under extreme conditions.

High-pressure NMR of proteins in solutions currently gains increasing interest. 3D structure determination of proteins under high pressure is, however, so far impossible due to the lack of NOE information. Residual dipolar couplings induced by the addition of magnetically orienting media are known to be capable of replacing NOE information to a very high extent. In the present contribution we study the pressure-stability of dimyristoylphosphatidylcholine (DMPC)/dihexanoylphosphatidylcholine (DHPC) bicelles and demonstrate the feasibility of measuring residual dipolar couplings in proteins under high pressure.

[Veterinary herd health consultancy on dairy farms: guidelines for starters]. / Veterinaire advisering van melkveebedrijven: een leidraad voor starters.

This paper presents a guideline which can be used when setting up a professional veterinary herd health and production management advisory programme for dairy farms. Earlier research showed that dairy farmers prefer a structured professional programme and ask their veterinary surgeons to provide an optimal veterinary advisory programme for their dairy farms with a clear structure and contents, and well-planned activities. The guideline presented here should aid in providing the farmers with that clarity, structure, and planning. This should ultimately lead to a more professional implementation of veterinary advisory programmes.

Solution NMR structure of the cold-shock protein from the hyperthermophilic bacterium Thermotoga maritima.

Cold-shock proteins (Csps) are a subgroup of the cold-induced proteins preferentially expressed in bacteria and other organisms on reduction of the growth temperature below the physiological temperature. They are related to the cold-shock domain found in eukaryotes and are some of the most conserved proteins known. Their exact function is still not known, but translational regulation, possibly via RNA chaperoning, has been discussed. Here we present the structure of a hyperthermophilic member of the Csp family. The NMR solution structure of TmCsp from Thermotoga maritima, the hyperthermophilic member of this class of proteins, was solved on the basis of 1015 conformational constraints. It contains five beta strands combined in two antiparallel beta sheets making up a beta barrel structure, in which beta strands 1-4 are arranged in a Greek-key topology. The side chain of R2, which is exclusively found in thermophilic members of the Csp family, probably participates in a peripheral ion cluster involving residues D20, R2, E47 and K63, suggesting that the thermostability of TmCsp is based on the peripheral ion cluster around the side chain of R2.

RFAC, a program for automated NMR R-factor estimation.

A computer program (RFAC) has been developed, which allows the automated estimation of residual indices (R-factors) for protein NMR structures and gives a reliable measure for the quality of the structures. The R-factor calculation is based on the comparison of experimental and simulated 1H NOESY NMR spectra. The approach comprises an automatic peak picking and a Bayesian analysis of the data, followed by an automated structure based assignment of the NOESY spectra and the calculation of the R-factor. The major difference to previously published R-factor definitions is that we take the non-assigned experimental peaks into account as well. The number and the intensities of the non-assigned signals are an important measure for the quality of an NMR structure. It turns out that for different problems optimally adapted R-factors should be used which are defined in the paper. The program allows to compute a global R-factor, different R-factors for the intra residual NOEs, the inter residual NOEs, sequential NOEs, medium range NOEs and long range NOEs. Furthermore, R-factors can be calculated for various user defined parts of the molecule or it is possible to obtain a residue-by-residue R-factor. Another possibility is to sort the R-factors according to their corresponding distances. The summary of all these different R-factors should allow the user to judge the structure in detail. The new program has been successfully tested on two medium sized proteins, the cold shock protein (TmCsp) from Termotoga maritima and the histidine containing protein (HPr) from Staphylococcus carnosus. A comparison with a previously published R-factor definition shows that our approach is more sensitive to errors in the calculated structure.

Pressure-induced local unfolding of the Ras binding domain of RalGDS.

The reliable prediction of the precise three-dimensional structure of proteins from their amino acid sequence is a major, still unresolved problem in biochemistry. Pressure is a parameter that controls folding/unfolding transitions of proteins through the volume change DeltaV of the protein-solvent system. By varying the pressure from 30 to 2,000 bar we detected using 15N/ 1H 2D NMR spectroscopy a unique equilibrium unfolding intermediate I in the Ras binding domain of the Ral guanine nucleotide dissociation stimulator (Ral GDS). It is characterized by a local melting of specific structural elements near hydrophobic cavities while the overall folded structure is maintained.

Backbone dynamics of sequence specific recognition and binding by the yeast Pho4 bHLH domain probed by NMR.

Backbone dynamics of the basic/helix-loop-helix domain of Pho4 from Saccharomyces cerevisae have been probed by NMR techniques, in the absence of DNA, nonspecifically bound to DNA and bound to cognate DNA. Alpha proton chemical shift indices and nuclear Overhauser effect patterns were used to elucidate the secondary structure in these states. These secondary structures are compared to the co-crystal complex of Pho4 bound to a cognate DNA sequence (Shimizu T. Toumoto A, Ihara K, Shimizu M, Kyogou Y, Ogawa N, Oshima Y, Hakoshima T, 1997, EMBO J 15: 4689-4697). The dynamic information provides insight into the nature of this DNA binding domain as it progresses from free in solution to a specifically bound DNA complex. Relative to the unbound form, we show that formation of either the nonspecific and cognate DNA bound complexes involves a large change in conformation and backbone dynamics of the basic region. The nonspecific and cognate complexes, however, have nearly identical secondary structure and backbone dynamics. We also present evidence for conformational flexibility at a highly conserved glutamate basic region residue. These results are discussed in relation to the mechanism of sequence specific recognition and binding.

Determination of mean and standard deviation of dihedral angles.

Backbone torsional angles are a characteristic and useful parameter for the description and characterisation of protein structures determined by x-ray crystallography or NMR spectroscopy. For the comparison of an ensemble of three-dimensional structures the calculation of the statistical parameters mean and standard deviation would be very useful. However, they are not defined unambiguously for periodic quantities such as the dihedral angles. In this paper a plausible and unique definition of these parameters is introduced and a straightforward method for their calculation is given.

An attempt to control lungworm disease and parasitic gastroenteritis on commercial dairy farms by the use of an adaptation of Michel's dose-and-move system.

On seven dairy farms an attempt was made to control lungworm disease in calves by turnout on a pasture grazed earlier by cows, followed by a move to aftermath and ivermectin treatment 2 months later. Transmission of lungworm was observed on all farms. Lungworm disease occurred on four farms at treatment. Coughing re-occurred on three of these farms in some animals 2 months later. Owing to poor performance between turnout and treatment, weight gain was below the norm on the farm with the highest infections and most severe respiratory signs. On the other farms respiratory signs did not result in poor weight gain. Gastrointestinal nematode infections remained low on all farms. The conclusion is that this dose and move scheme cannot be recommended for the control of lungworm.