Balancing dairy cattle diets for rumen nitrogen and methionine or all essential amino acids relative to metabolizable energy.

Improving the ability of diet formulation models to more accurately predict AA supply while appropriately describing requirements for lactating dairy cattle provides an opportunity to improve animal productivity, reduce feed costs, and reduce N intake. The goal of this study was to evaluate the sensitivity of a new version of the Cornell Net Carbohydrate and Protein System (CNCPS) to formulate diets for rumen N, Met, and all essential AA (EAA). Sixty-four high-producing dairy cattle were randomly assigned to 1 of the 4 following diets in a 14-wk longitudinal study: (1) limited metabolizable protein (MP), Met, and rumen N (Base), (2) adequate Met but limited MP and rumen N (Base + M), (3) adequate Met and rumen N, but limited MP (Base + MU), and (4) adequate MP, rumen N, and balanced for all EAA (Positive). All diets were balanced to exceed requirements for ME relative to maintenance and production, assuming a nonpregnant, 650-kg animal producing 40 kg of milk at 3.05% true protein and 4.0% fat. Dietary MP was 97.2, 97.5, 102.3, and 114.1 g/kg of dry matter intake for the Base, Base + M, Base + MU, and Positive diets, respectively. Differences were observed for dry matter intake and milk yield (24.1 to 24.7 and 39.4 to 41.1 kg/d, among treatments). Energy corrected milk, fat, and true protein yield were greater (2.9, 0.13, and 0.08 kg/d, respectively) in cows fed the Positive compared with the Base diet. Using the updated CNCPS, cattle fed the Base, Base + M, and Base + MU diets were predicted to have a negative MP balance (-231, -310, and -142 g/d, respectively), whereas cattle fed the Positive diet consumed 33 g of MP/d excess to ME supply. Bacterial growth was predicted to be depressed by 16 and 17% relative to adequate N supply for the Base and Base + M diets, respectively, which corresponded with the measured lower apparent total-tract NDF degradation. The study demonstrates that improvements in lactation performances can be achieved when rumen N and Met are properly supplied and further improved when EAA supply are balanced relative to requirements. Formulation using the revised CNCPS provided predictions for these diets, which were sensitive to changes in rumen N, Met, all EAA, and by extension MP supply.

The Cornell Net Carbohydrate and Protein System: Updates to the model and evaluation of version 6.5.

New laboratory and animal sampling methods and data have been generated over the last 10 yr that had the potential to improve the predictions for energy, protein, and AA supply and requirements in the Cornell Net Carbohydrate and Protein System (CNCPS). The objectives of this study were to describe updates to the CNCPS and evaluate model performance against both literature and on-farm data. The changes to the feed library were significant and are reported in a separate manuscript. Degradation rates of protein and carbohydrate fractions were adjusted according to new fractionation schemes, and corresponding changes to equations used to calculate rumen outflows and postrumen digestion were presented. In response to the feed-library changes and an increased supply of essential AA because of updated contents of AA, a combined efficiency of use was adopted in place of separate calculations for maintenance and lactation to better represent the biology of the cow. Four different data sets were developed to evaluate Lys and Met requirements, rumen N balance, and milk yield predictions. In total 99 peer-reviewed studies with 389 treatments and 15 regional farms with 50 different diets were included. The broken-line model with plateau was used to identify the concentration of Lys and Met that maximizes milk protein yield and content. Results suggested concentrations of 7.00 and 2.60% of metabolizable protein (MP) for Lys and Met, respectively, for maximal protein yield and 6.77 and 2.85% of MP for Lys and Met, respectively, for maximal protein content. Updated AA concentrations were numerically higher for Lys and 11 to 18% higher for Met compared with CNCPS v6.0, and this is attributed to the increased content of Met and Lys in feeds that were previously incorrectly analyzed and described. The prediction of postruminal flows of N and milk yield were evaluated using the correlation coefficient from the BLUP (R(2)BLUP) procedure or model predictions (R(2)MDP) and the concordance correlation coefficient. The accuracy and precision of rumen-degradable N and undegradable N and bacterial N flows were improved with reduced bias. The CNCPS v6.5 predicted accurate and precise milk yield according to the first-limiting nutrient (MP or metabolizable energy) with a R(2)BLUP=0.97, R(2)MDP=0.78, and concordance correlation coefficient=0.83. Furthermore, MP-allowable milk was predicted with greater precision than metabolizable energy-allowable milk (R(2)MDP=0.82 and 0.76, respectively, for MP and metabolizable energy). Results suggest a significant improvement of the model, especially under conditions of MP limitation.

Updating the Cornell Net Carbohydrate and Protein System feed library and analyzing model sensitivity to feed inputs.

The Cornell Net Carbohydrate and Protein System (CNCPS) is a nutritional model that evaluates the environmental and nutritional resources available in an animal production system and enables the formulation of diets that closely match the predicted animal requirements. The model includes a library of approximately 800 different ingredients that provide the platform for describing the chemical composition of the diet to be formulated. Each feed in the feed library was evaluated against data from 2 commercial laboratories and updated when required to enable more precise predictions of dietary energy and protein supply. A multistep approach was developed to predict uncertain values using linear regression, matrix regression, and optimization. The approach provided an efficient and repeatable way of evaluating and refining the composition of a large number of different feeds against commercially generated data similar to that used by CNCPS users on a daily basis. The protein A fraction in the CNCPS, formerly classified as nonprotein nitrogen, was reclassified to ammonia for ease and availability of analysis and to provide a better prediction of the contribution of metabolizable protein from free AA and small peptides. Amino acid profiles were updated using contemporary data sets and now represent the profile of AA in the whole feed rather than the insoluble residue. Model sensitivity to variation in feed library inputs was investigated using Monte Carlo simulation. Results showed the prediction of metabolizable energy was most sensitive to variation in feed chemistry and fractionation, whereas predictions of metabolizable protein were most sensitive to variation in digestion rates. Regular laboratory analysis of samples taken on-farm remains the recommended approach to characterizing the chemical components of feeds in a ration. However, updates to the CNCPS feed library provide a database of ingredients that are consistent with current feed chemistry information and laboratory methods and can be used as a platform to formulate rations and improve the description of biology within the model.

A novel TLR2 agonist from Bordetella pertussis is a potent adjuvant that promotes protective immunity with an acellular pertussis vaccine.

Bordetella pertussis causes whooping cough, a severe and often lethal respiratory infection in infants. A recent resurgence of pertussis has been linked with waning or suboptimal immunity induced with acellular pertussis vaccines (Pa) that were introduced to most developed countries in the 1990s because of safety concerns around the use of whole-cell pertussis vaccines (Pw). Pa are composed of individual B. pertussis antigens absorbed to alum and promote strong antibody, T helper type 2 (Th2) and Th17 responses, but are less effective at inducing cellular immunity mediated by Th1 cells. In contrast, Pw, which include endogenous Toll-like receptor (TLR) agonists, induce Th1 as well as Th17 responses. Here we report the identification and characterization of novel TLR2-activating lipoproteins from B. pertussis. These proteins contain a characteristic N-terminal signal peptide that is unique to Gram-negative bacteria and we demonstrate that one of these lipoproteins, BP1569, activates murine dendritic cells and macrophages and human mononuclear cells via TLR2. Furthermore, we demonstrated that a corresponding synthetic lipopeptide LP1569 has potent immunostimulatory and adjuvant properties, capable of enhancing Th1, Th17, and IgG2a antibody responses induced in mice with an experimental Pa that conferred superior protection against B. pertussis infection than an equivalent vaccine formulated with alum.

M1 and m2 muscarinic receptor subtypes regulate antidepressant-like effects of the rapidly acting antidepressant scopolamine.

Scopolamine produces rapid and significant symptom improvement in patients with depression, and most notably in patients who do not respond to current antidepressant treatments. Scopolamine is a nonselective muscarinic acetylcholine receptor antagonist, and it is not known which one or more of the five receptor subtypes in the muscarinic family are mediating these therapeutic effects. We used the mouse forced-swim test, an antidepressant detecting assay, in wild-type and transgenic mice in which each muscarinic receptor subtype had been genetically deleted to define the relevant receptor subtypes. Only the M1 and M2 knockout (KO) mice had a blunted response to scopolamine in the forced-swim assay. In contrast, the effects of the tricyclic antidepressant imipramine were not significantly altered by gene deletion of any of the five muscarinic receptors. The muscarinic antagonists biperiden, pirenzepine, and VU0255035 (N-[3-oxo-3-[4-(4-pyridinyl)-1-piper azinyl]propyl]-2,1,3-benzothiadiazole-4-sulfonamide) with selectivity for M1 over M2 receptors also demonstrated activity in the forced-swim test, which was attenuated in M1 but not M2 receptor KO mice. An antagonist with selectivity of M2 over M1 receptors (SCH226206 [(2-amino-3-methyl-phenyl)-[4-[4-[[4-(3 chlorophenyl)sulfonylphenyl]methyl]-1-piperidyl]-1-piperidyl]methanone]) was also active in the forced-swim assay, and the effects were deleted in M2 (-/-) mice. Brain exposure and locomotor activity in the KO mice demonstrated that these behavioral effects of scopolamine are pharmacodynamic in nature. These data establish muscarinic M1 and M2 receptors as sufficient to generate behavioral effects consistent with an antidepressant phenotype and therefore as potential targets in the antidepressant effects of scopolamine.

Evaluation of various static in vitro-in vivo extrapolation models for risk assessment of the CYP3A inhibition potential of an investigational drug.

Nine static models (seven basic and two mechanistic) and their respective cutoff values used for predicting cytochrome P450 3A (CYP3A) inhibition, as recommended by the US Food and Drug Administration and the European Medicines Agency, were evaluated using data from 119 clinical studies with orally administered midazolam as a substrate. Positive predictive error (PPE) and negative predictive error (NPE) rates were used to assess model performance, based on a cutoff of 1.25-fold change in midazolam area under the curve (AUC) by inhibitor. For reversible inhibition, basic models using total or unbound systemic inhibitor concentration [I] had high NPE rates (46-47%), whereas those using intestinal luminal ([I]gut) values had no NPE but a higher PPE. All basic models for time-dependent inhibition had no NPE and reasonable PPE rates (15-18%). Mechanistic static models that incorporate all interaction mechanisms and organ specific [I] values (enterocyte and hepatic inlet) provided a higher predictive precision, a slightly increased NPE, and a reasonable PPE. Various cutoffs for predicting the likelihood of CYP3A inhibition were evaluated for mechanistic models, and a cutoff of 1.25-fold change in midazolam AUC appears appropriate.

The effect of starch-, fiber-, or sugar-based supplements on nitrogen utilization in grazing dairy cows.

Nitrogen utilization in grazing cows is often low due to high concentrations of rapidly soluble and degradable protein in the pasture-based diet. Broadly, opportunities to improve N utilization lie in either reducing the amount of N consumed by the animal, or incorporating more N into milk protein. The goal of this study was to compare the relative importance of dietary N intake and productive N output for improving N utilization in grazing cows fed either starch-, fiber-, or sugar-based supplements. Also, the Cornell Net Carbohydrate and Protein System (CNCPS; Cornell University, Ithaca, NY) was evaluated as a tool to assess cow performance and improve N utilization in pasture-based systems. Eighty-five cows were randomly assigned to 1 of 5 treatments at parturition (17 cows per treatment). Treatments consisted of a pasture-only control and pasture with a starch- (St and StN), fiber- (FbN), or a sugar-based supplement. The StN and FbN treatments contained additional dietary N. Diets were formulated using the CNCPS to supply similar levels of dietary metabolizable energy, but differing levels of dietary N and metabolizable protein. Nitrogen utilization ranged from 22 to 26% across the 5 groups. Cows fed the St diet had the lowest levels of milk urea N, blood urea N, and urinary N excretion and had the highest productive N output (149 g/d). Cows fed the FbN treatment had similar productive N output (137 g/d) and consumed approximately 100g/d more dietary N than the St treatment, resulting in greater urinary N excretion. Although milk protein yield was moderately greater in the St treatment, quantitatively the difference in N intake (100g/d) had the greatest effect on N utilization and suggests that controlling dietary N intake should be the first priority when attempting to improve N utilization in grazing cows. No effect was observed of supplementing pasture-fed cattle with sugar on production or N utilization under the conditions of this experiment. Predictions of metabolizable energy and protein availability for milk yield from the CNCPS were similar to actual milk yield for all treatments. Model-predicted N utilization and excretion reflected the trends observed in the measured data and suggests that the CNCPS can be a useful tool for formulating and evaluating diets to improve N utilization in pasture-based systems.

Immunity to the respiratory pathogen Bordetella pertussis.

Bordetella pertussis causes whooping cough, a severe respiratory tract infection in infants and children, and also infects adults. Studies in murine models have shown that innate immune mechanisms involving dendritic cells, macrophages, neutrophils, natural killer cells, and antimicrobial peptides help to control the infection, while complete bacterial clearance requires cellular immunity mediated by T-helper type 1 (Th1) and Th17 cells. Whole cell pertussis vaccines (wP) are effective, but reactogenic, and have been replaced in most developed countries by acellular pertussis vaccines (aP). However, the incidence of pertussis is still high in many vaccinated populations; this may reflect sub-optimal, waning, or escape from immunity induced by current aP. Protective immunity generated by wP appears to be mediated largely by Th1 cells, whereas less efficacious alum-adjuvanted aP induce strong antibody Th2 and Th17 responses. New generation aP that induce Th1 rather than Th2 responses are required to improve vaccine efficacy and prevent further spread of B. pertussis.

Development and evaluation of equations in the Cornell Net Carbohydrate and Protein System to predict nitrogen excretion in lactating dairy cows.

Nitrogen excretion is of particular concern on dairy farms, not only because of its effects on water quality, but also because of the subsequent release of gases such as ammonia to the atmosphere. To manage N excretion, accurate estimates of urinary N (UN) and fecal N (FN) are needed. On commercial farms, directly measuring UN and FN is impractical, meaning that quantification must be based on predictions rather than measured data. The purpose of this study was to use a statistical approach to develop equations and evaluate the Cornell Net Carbohydrate and Protein System's (CNCPS) ability to predict N excretion in lactating dairy cows, and to compare CNCPS predictions to other equations in the literature. Urinary N was over-predicted by the CNCPS due to inconsistencies in N accounting within the model that partitioned more N to feces than urine, although predicted total N excretion was reasonable. Data to refine model predictions were compiled from published studies (n=32) that reported total collection N balance results. Considerable care was taken to ensure the data included in the development data set (n=104) accounted for >90% of the N intake (NI). Unaccounted N for the compiled data set was 1.47±4.60% (mean ± SD). The results showed that FN predictions could be improved by using a modification of a previously published equation: FN (g/d) = [[NI (g/kg of organic matter) × (1 - 0.842)] + 4.3 × organic matter intake (kg/d)] × 1.20, which, when evaluated against the compiled N balance data, had a squared coefficient of determination based on a mean study effect R(MP)(2) of 0.73, concurrent correlation coefficient (CCC) of 0.83 and a root mean square error (RMSE) of 10.38 g/d. Urinary N is calculated in the CNCPS as the difference between NI and other N excretion and losses. Incorporating the more accurate FN prediction into the current CNCPS framework and correcting an internal calculation error considerably improved UN predictions (RMSE=12.73 g/d, R(MP)(2)=0.86, CCC=0.90). The changes to FN and UN translated into an improved prediction of total manure N excretion (RMSE=12.42 g/d, R(MP)(2)=0.96, CCC=0.97) and allows nutritionists and farm advisors to evaluate these factors during the ration formulation process.

Interfacial fracture toughness between bovine cortical bone and cements.

To evaluate the bonding strength of the interfaces within the cemented arthroplasty system, various mechanical tests have been used. Conventional push-out and pull-out tests cannot reveal the actual bonding property of the interface because of the significant influence of surface roughness on the measured adhesion and the failure to account for the mismatch of elastic modulus across the interface. An alternative fracture mechanics approach, which considers the mix of opening and shear modes of the crack tip loading associated with the testing system and the elastic mismatch of materials across the interface, was used to evaluate the bonding ability of various cements. The four-point bend interfacial delamination test by Charalambides et al. (J. Appl. Mech. 56 (1989) 77; Mech. Mater. 8 (1990) 269) was used to quantify the bonding ability of cements. This method is arguably more suitable since the applied loading mode is comparable to the nature of loading within the prosthetic system, which is primarily bending. The bovine bone specimens were polished to mirror finish to eliminate bonding by mechanical interlocking. The results revealed minimal bonding for the conventional bone cement (PMMA) whereas substantial bonding was evident for the glass-ionomer cements tested. However, only the conventional glass-ionomer cements showed evidence of bonding on testing, while the resin-modified glass-ionomer cement (poly-HEMA) did not. The latter appeared to debond before testing because of excessive expansion stresses associated with swelling in water.

Comparison of failure characteristics of a range of cancellous bone-bone cement composites.

Over the past decade, orthopedic surgery has embraced an increase in the depth of cement penetration into the adjacent cancellous bone structure. The resultant interdigitation transforms this zone into a thick layer of continuous interpenetrating composite material. The failure behavior of the composite formed with a number of potential bone cements with different bonding ability was investigated. The cancellous bone-cement composites exhibit considerable resistance to crack extension, and in situ optical observation indicates that the contribution of the cancellous bone is analogous to that of a typical fiber bridging process. The critical stress intensity factor and the work of fracture have been used to quantify the failure characteristics of the cancellous bone-cement composites. The nature of the crack propagation through these cement-bone composites was also captured via optical microscopy, and scanning electron microscopic images were taken of the failure surfaces. The R-curve behavior, or crack extension characteristic, of the cancellous bone-cement composite was also determined. The interesting outcome is that the cancellous bone-PMMA (poly-methylmethacrylate) composite, despite the absence of chemical bonding with bone, required the highest energy to fracture. In addition, the dimensional stability of the cement has a great effect on the interface.

Time dependence of the mechanical properties of GICs in simulated physiological conditions.

The mechanical properties of glass-ionomer cements (GICs) have been satisfactory for dental applications and have shown their potential in orthopedic surgery. Because the physiological environment in orthopedics is different from dentistry by unavoidable contamination with blood and other fluids such as normal saline used during an operation, the determination of GICs for orthopedic applications should be performed in an appropriate environment. The properties of a novel resin-modified GIC, S430, for orthopedic applications were evaluated in simulated orthopedic conditions by an early exposure to and long-term storage in normal saline. An early exposure to normal saline caused 20-60% reduction of its compressive and flexural properties, whereas long-term storage in normal saline showed slight changes of its mechanical properties. The effects were probably due to the disturbance of the cross-linking formation in the acid-base reaction and also the reduction of electrostatic interactions of the cross-linking polymeric chain of hydroxyethyl methacrylate (HEMA) in resin-modified GIC.

Effects of glass ionomer cements on bone tissue.

In vivo biocompatibility of glass ionomer cements (GICs) was evaluated for use in orthopaedic surgery using a rat model and compared with conventional bone cement, Polymethyl methacrylate, PMMA. The unset GICs and PMMA were inserted into the marrow cavities of rat femora and retained in situ for various periods of time. The PMMA bone cement showed complete biocompatibility with no interference with reparative bone. The conventional GIC with smaller glass particles and lower powder/liquid ratio showed an initial minor toxic effect on rat bone tissue with later disturbance of adjacent bone formation. The conventional GIC with larger-size glass particles and higher powder/liquid ratio and resin-modified GIC showed more severe toxic effect on rat tissue with the resin-modified GIC affecting the rat bone tissue later. The causes of toxicity associated with the conventional GIC with larger glass particles and higher powder/liquid ration and the resin-modified GIC are thought to be related with the unreacted acid component of both materials and longer ongoing metallic ion release.

Fracture toughness of bovine bone: influence of orientation and storage media.

Using the single-edge notched bending (SENB) test, two fracture toughness parameters of longitudinal and transverse bovine bone specimens were evaluated: the critical stress intensity factor, Kc, determined from the peak load to initiate fracture, and the energy or work of fracture, Wf, the energy required to extend a crack through a notched specimen. It was found that preservation of bone in alcohol resulted in a 25-45% higher Kc value compared to control specimens stored in physiological saline; whereas the work of fracture, Wf, demonstrated the opposite behaviour, with the alcohol stored specimens having a 28-56% lower value than the saline control specimens. It was established that the effect of alcohol is reversible upon the bone being restored in saline. Consistent with previous studies, it was found that cracks oriented in the longitudinal direction resulted in both a significantly lower fracture toughness and lower work of fracture than those cracks directed transversely. The results are discussed in terms of the proposed deformation and fracture mechanisms known to occur in bone.

GPs' views of discussions of prognosis in severe COPD.

BACKGROUND: Modern palliative care promotes open communication between doctor and patient, which includes access to information about prognosis. GPs play a major role in managing chronic obstructive pulmonary disease (COPD) patients in the final stages of illness. Their views of discussions of prognosis are therefore important if the principles of palliative care are to be extended to COPD. OBJECTIVE: Our aim was to investigate the role that discussions of prognosis play in GPs' management of patients with severe COPD and the factors that influence those discussions. METHODS: We conducted a questionnaire survey of all GP principals of one inner London Health Authority (n = 389) in April 1999. Questionnaire development involved a literature review to identify issues of importance to GPs in the discussion of prognosis in COPD, and in-depth interviews with five GPs. RESULTS: Of the 214 respondents (55% response), 72.5% thought that discussions of prognosis were often necessary or essential in severe COPD. The majority (82%) felt that GPs have an important role in these discussions. However, only a minority (41%) of GPs reported often or always discussing prognosis. Half the GPs were undecided as to whether most patients with COPD wanted to know about their prognosis. Among the GPs who reported rarely or never discussing prognosis (n = 33), a majority felt ill-prepared to discuss the subject (60% reported that there was insufficient information in the primary care notes to be able to discuss prognosis, and 64% found it hard to start discussions with patients). CONCLUSION: Although the majority of GPs acknowledged a need to discuss prognosis in severe COPD, this was not reflected in their reported behaviour. It appears that the palliative care approach of open communication, whilst seen to be relevant to severe COPD, is not applied routinely in managing the disease in primary care. Uncertainty among GPs as to how patients view the discussion of prognosis and inadequate preparation may pose potential barriers.

Evaluating acrylic and glass-ionomer cement strength using the biaxial flexure test.

Frequently, bone cement strengths are evaluated from uniaxial tests, such as three- or four-point flexure. Measurement of material strength in this manner may not provide an accurate characterisation of a bone cements true load-bearing capacity. In most orthopaedic applications, there exists a state of biaxial stress and so biaxial strength information is most useful. To address this issue, the biaxial flexure strength of two polymethylmethacrylate orthopaedic (PMMA) bone cements and two glass-ionomer dental cements have been determined. The biaxial strength of orthopaedic bone cements have been compared to the three-point bending strength. Furthermore, the calculated theoretical biaxial strength was compared with a value of biaxial strength utilising the finite element method. For all materials tested the calculated biaxial strength is significantly greater than the three-point bending strength. The biaxial test offers several advantages over three-point bending because it critically explores surface flaws--as it does not matter in which orientation the crack lies. However, it does minimise the volume or surface area investigated and also the edge effect. The difference in strength calculated for the two testing methods can be explained quantitatively in terms of the volume of material under stress. This work has demonstrated that the biaxial flexure test can be used for the testing of orthopaedic bone cements.

Teaching professional development in medical schools.

Doctors must increasingly be aware of what they should be, as well as what they should know. Professionalism, including a value system that supports the compassionate care of patients, is a means of encapsulating and prioritising these competing responsibilities. Accordingly, in this article, we assume that professionalism is an essential aspect of medical practice that needs to be taught to those entering medicine. We first describe critiques of professionalism and current challenges to it, in practice and in medical education. We then assess the current efforts of curriculum reform to incorporate professionalism and the methods used to teach it. Adopting and assessing such approaches to ensure that they are effective is of central importance in the education of future clinicians.

Changing face of medical curricula.

The changing role of medicine in society and the growing expectations patients have of their doctors means that the content and delivery of medical curricula also have to change. The focus of health care has shifted from episodic care of individuals in hospitals to promotion of health in the community, and from paternalism and anecdotal care to negotiated management based on evidence of effectiveness and safety. Medical training is becoming more student centred, with an emphasis on active learning rather than on the passive acquisition of knowledge, and on the assessment of clinical competence rather than on the ability to retain and recall unrelated facts. Rigid educational programmes are giving way to more adaptable and flexible ones, in which student feedback and patient participation have increasingly important roles. The implementation of sustained innovation in medical education continues to present challenges, especially in terms of providing institutional and individual incentives. However, a continuously evolving, high quality medical education system is needed to assure the continued delivery of high quality medicine.