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BACKGROUND AND AIMS: HCV infection continues to be a major global health burden despite effective antiviral treatments. The urgent need for a protective vaccine is hindered by the scarcity of suitable HCV-permissive animal models tractable in vaccination and challenge studies. Currently, only antibody neutralization studies in infectious cell culture systems or studies of protection by passive immunization of human liver chimeric mice offer the possibility to evaluate the effect of vaccine-induced antibodies. However, differences between culture-permissive and in vivo-permissive viruses make it a challenge to compare analyses between platforms. To address this problem, we aimed at developing genotype-specific virus variants with genetic stability both in vitro and in vivo. APPROACH AND RESULTS: We demonstrated infection of human liver chimeric mice with cell culture-adapted HCV JFH1-based Core-NS2 recombinants of genotype 1-6, with a panel of 10 virus strains used extensively in neutralization and receptor studies. Clonal re-engineering of mouse-selected mutations resulted in virus variants with robust replication both in Huh7.5 cells and human liver chimeric mice, with genetic stability. Furthermore, we showed that, overall, these virus variants have similar in vitro neutralization profiles as their parent strains and demonstrated their use for in vivo neutralization studies. CONCLUSIONS: These mouse-selected HCV recombinants enable the triage of new vaccine-relevant antibodies in vitro and further allow characterization of protection from infection in vivo using identical viruses in human liver chimeric mice. As such, these viruses will serve as important resources in testing novel antibodies and can thus guide strategies to develop an efficient protective vaccine against HCV infection.

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OBJECTIVE: To explore the relationships between leader support, staff influence over decisions, work pressure and patient satisfaction. DESIGN: A cross-sectional study of large National Health Service (NHS) datasets in England in 2010. SETTING AND PARTICIPANTS: 158 NHS acute hospital trusts in England (n=63 156) from all staff groups. PRIMARY AND SECONDARY OUTCOME MEASURES: Survey data measuring leader support, staff influence over decision making, staff work pressure and objective outcome data measuring patient satisfaction. RESULTS: Multilevel serial mediation analysis showed a significantly positive association between leader support and staff influence over decisions (B=0.74, SE=0.07, p<0.01). Furthermore, staff influence over decisions showed a negative association with staff work pressure (B=-0.84, SE=0.41, p<0.05) which in turn was negatively linked to patient satisfaction (B=-17.50, SE=4.34, p<0.01). Serial mediation showed a positive indirect effect of leader support on patient satisfaction via staff influence over decisions and work pressure (B=10.96, SE=5.55, p<0.05). CONCLUSIONS AND IMPLICATIONS: Our results provide evidence that leader support influences patient satisfaction through shaping staff experience, particularly staff influence over decisions and work pressure. Patients' care is dependent on the health, well-being, and effectiveness of the NHS workforce. That, in turn, is determined by the extent to which leaders are supportive in ensuring that work environments are managed in a way which protects the well-being of staff.

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Combustion of coal for energy generation has been a significant contributor to increased concentrations of atmospheric carbon dioxide. It is of interest to evaluate the potential of former coalfields for mitigating these increases by carbon sequestration and to compare different options to achieving this end. Here, carbon sequestration in residual coal seams and through reclamation of spoil tips is compared, and their carbon dioxide storage potential in the South Wales Coalfield estimated. Coal seam sequestration estimates come from an established methodology and consider the total unmined coal resource below 500 m deep with potential for carbon sequestration. The most likely effective deep seam storage capacity is 104.9 Mt carbon dioxide, taking account of reservoir conditions and engineering factors. Whilst many spoil tips in South Wales have been reclaimed, the focus has not been on carbon sequestration potential. Estimates of minesoil restoration sequestration capacity were based on a survey of restored minesoil and vegetation carbon stocks, mainly on sites 20-30 years after restoration; data from this survey were then extrapolated to the coalfield as a whole. Minesoil storage is estimated at 1.5 or 2.5 Mt (+2.2 Mt in tree biomass) carbon dioxide based on average grassland or woodland measurements, respectively; modelled data predicted equilibrium values of 2.9 and 2.6 Mt carbon dioxide respectively in grassland or woodland minesoils. If all sites achieved close to the maximum capacity in their land use class, minesoil storage capacity would increase to 2.1 or 3.9 Mt carbon dioxide, respectively. Combining the best woodland minesoil and standing biomass values, sequestration capacity increases to 7.2 Mt carbon dioxide. The wider social, economic, environmental and regulatory constraints to achieving this sequestration for each approach are discussed. Coal seam sequestration has a much higher capacity but sequestration in mine sites is less costly and has fewer regulatory constraints. Findings indicate a significant combined potential for carbon sequestration in the South Wales Coalfield and highlight challenges in achieving this potential. On a global scale, ex-coalfield sequestration could contribute to broader efforts to mitigate emissions.

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BACKGROUND & AIMS: Inhibitors of the hepatitis C virus (HCV) NS5A protein are a key component of effective treatment regimens, but the genetic heterogeneity of HCV has limited the efficacy of these agents and mutations lead to resistance. We directly compared the efficacy of all clinically relevant NS5A inhibitors against HCV genotype 1-7 prototype isolates and resistant escape variants, and investigated the effects of pre-existing resistance-associated substitutions (RAS) on HCV escape from treatment. METHODS: We measured the efficacy of different concentrations of daclatasvir, ledipasvir, ombitasvir, elbasvir, ruzasvir, velpatasvir, and pibrentasvir in cultured cells infected with HCV recombinants expressing genotype 1-7 NS5A proteins with or without RAS. We engineered HCV variants that included RAS identified in escape experiments, using recombinants with or without T/Y93H and daclatasvir, or that contained RAS previously reported from patients. RESULTS: NS5A inhibitors had varying levels of efficacy against original and resistant viruses. Only velpatasvir and pibrentasvir had uniform high activity against all HCV genotypes tested. RAS hotspots in NS5A were found at amino acids 28, 30, 31, and 93. Engineered escape variants had high levels of fitness. Pibrentasvir had the highest level of efficacy against variants; viruses with RAS at amino acids 28, 30, or 31 had no apparent resistance to pibrentasvir, and HCV with RAS at amino acid 93 had a low level of resistance to this drug. However, specific combinations of RAS and deletion of amino acid 32 led to significant resistance to pibrentasvir. For the remaining NS5A inhibitors tested, RAS at amino acids 28 and 93 led to high levels of resistance. Among these inhibitors, velpatasvir was more effective against variants with RAS at amino acid 30 and some variants with RAS at amino acid 31 than the other agents. Variants with the pre-existing RAS T/Y93H acquired additional NS5A changes during escape experiments, resulting in HCV variants with specific combinations of RAS, showing high fitness and high resistance. CONCLUSIONS: We performed a comprehensive comparison of the efficacy of the 7 clinically relevant inhibitors of HCV NS5A and identified variants associated with resistance to each agent. These findings could improve treatment of patients with HCV infection.

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Hepatitis C virus (HCV) strains belong to seven genotypes with numerous subtypes that respond differently to antiviral therapies. Genotype 1, and primarily subtype 1b, is the most prevalent genotype worldwide. The development of recombinant HCV infectious cell culture systems for different variants, permitted by the high replication capacity of strain JFH1 (genotype 2a), has advanced efficacy and resistance testing of antivirals. However, efficient infectious JFH1-based cell cultures of subtype 1b are limited and comprise only the 5' untranslated region (5'UTR)-NS2, NS4A, or NS5A regions. Importantly, it has not been possible to develop efficient 1b infectious systems expressing the NS3/4A protease, an important target of direct-acting antivirals. We developed efficient infectious JFH1-based cultures with genotype 1b core-NS5A sequences of strains DH1, Con1, and J4 by using previously identified HCV cell culture adaptive substitutions A1226G, R1496L, and Q1773H. These viruses spread efficiently in Huh7.5 cells by acquiring additional adaptive substitutions, and final recombinants yielded peak supernatant infectivity titers of 4 to 5 log10 focus-forming units (FFU)/ml. We subsequently succeeded in adapting a JFH1-based 5'UTR-NS5A DH1 recombinant to efficient growth in cell culture. We evaluated the efficacy of clinically relevant NS3/4A protease and NS5A inhibitors against the novel genotype 1b viruses, as well as against previously developed 1a viruses. The inhibitors were efficient against all tested genotype 1 viruses, with NS5A inhibitors showing half-maximal effective concentrations several orders of magnitude lower than NS3/4A protease inhibitors. In summary, the developed HCV genotype 1b culture systems represent valuable tools for assessing the efficacy of various classes of antivirals and for other virological studies requiring genotype 1b infectious viruses.

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UNLABELLED: Human monoclonal antibodies (HMAbs) with neutralizing capabilities constitute potential immune-based treatments or prophylaxis against hepatitis C virus (HCV). However, lack of cell culture-derived HCV (HCVcc) harboring authentic envelope proteins (E1/E2) has hindered neutralization investigations across genotypes, subtypes, and isolates. We investigated the breadth of neutralization of 10 HMAbs with therapeutic potential against a panel of 16 JFH1-based HCVcc-expressing patient-derived Core-NS2 from genotypes 1a (strains H77, TN, and DH6), 1b (J4, DH1, and DH5), 2a (J6, JFH1, and T9), 2b (J8, DH8, and DH10), 2c (S83), and 3a (S52, DBN, and DH11). Virus stocks used for in vitro neutralization analysis contained authentic E1/E2, with the exception of full-length JFH1 that acquired the N417S substitution in E2. The 50% inhibition concentration (IC50) for each HMAb against the HCVcc panel was determined by dose-response neutralization assays in Huh7.5 cells with antibody concentrations ranging from 0.0012 to 100 µg/mL. Interestingly, IC50 values against the different HCVcc's exhibited large variations among the HMAbs, and only three HMAbs (HC-1AM, HC84.24, and AR4A) neutralized all 16 HCVcc recombinants. Furthermore, the IC50 values for a given HMAb varied greatly with the HCVcc strain, which supports the use of a diverse virus panel. In cooperation analyses, HMAbs HC84.24, AR3A, and, especially HC84.26, demonstrated synergistic effects towards the majority of the HCVcc's when combined individually with AR4A. CONCLUSION: Through a neutralization analysis of 10 clinically relevant HMAbs against 16 JFH1-based Core-NS2 recombinants from genotypes 1a, 1b, 2a, 2b, 2c, and 3a, we identified at least three HMAbs with potent and broad neutralization potential. The neutralization synergism obtained when pooling the most potent HMAbs could have significant implications for developing novel strategies to treat and control HCV.

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UNLABELLED: Immunotherapy and vaccine development for hepatitis C virus (HCV) will depend on broadly reactive neutralizing antibodies (NAbs). However, studies in infectious strain JFH1-based culture systems expressing patient-derived Core-NS2 proteins have suggested neutralization resistance for specific HCV strains, in particular, of genotype 2. To further examine this phenomenon, we developed a panel of HCV genotype 2 recombinants for testing of sensitivity to neutralization by chronic-phase patient sera and lead human monoclonal antibodies (HMAbs). The novel Core-NS2 recombinants, with patient-derived genotype 2a (strain T9), 2b (strains DH8 and DH10), and 2c (strain S83) consensus sequences, were viable in Huh7.5 hepatoma cells without requirement for adaptive mutations, reaching HCV infectivity titers of 3.9-4.5 log10 focus-forming units per milliliter. In in vitro neutralization assays, we demonstrated that the novel genotype 2 viruses as well as prototype strains J6/JFH1(2a) and J8/JFH1(2b), all with authentic envelope proteins, were resistant to neutralization by genotype 2a, 2b, 2c, 2j, 2i, and 2q patient sera. However, these patient sera had high titers of HCV-specific NAbs, because they efficiently reduced the infectivity of J6(2a) and J8(2b) with deleted hypervariable region 1. The genotype 2a, 2b, and 2c viruses, found resistant to polyclonal patient sera neutralization, were efficiently neutralized by two lead HMAbs (AR4A and HC84.26). CONCLUSION: Using novel 2a, 2b, and 2c cell-culture systems, expressing authentic envelope proteins, we demonstrated resistance of HCV to patient-derived polyclonal high-titer NAbs. However, the same genotype 2 culture viruses were all sensitive to HMAbs recognizing conformational epitopes, indicating that neutralization resistance of HCV can be overcome by applying recombinant antibodies. These findings have important implications for HCV immunotherapy and vaccine development.

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The correlation of neutralizing antibodies to treatment outcome in patients with chronic hepatitis C virus (HCV) infection has not been established. The aim of this study was to determine whether neutralizing antibodies could be used as an outcome predictor in patients with chronic HCV, genotype 1, infection treated with pegylated interferon-α and ribavirin. Thirty-nine patients with chronic hepatitis C, genotype 1a or 1b, with either sustained virologic response (n = 23) or non-sustained virologic response (n = 16) were enrolled. Samples taken prior to treatment were tested for their ability to neutralize 6 different HCV genotype 1 cell culture recombinants (1a: H77/JFH1, TN/JFH1, DH6/JFH1; 1b: J4/JFH1, DH1/JFH1, DH5/JFH1). The results were expressed as the highest dilution yielding 50% neutralization (NAb50-titer). We observed no genotype or subtype specific differences in NAb50-titers between patients with chronic HCV infection with and without sustained virologic response when tested against any of the included culture viruses. However, NAb50-titers varied significantly with a mean reciprocal NAb50-titer of 800 (range: 100-6400) against DH6/JFH1 compared to a mean NAb50-titer of 50 (range: <50-400) against all other included isolates. Subsequent studies demonstrated that the efficient neutralization of DH6/JFH1 could be linked to engineered adaptive mutations in the envelope-2 protein. In analysis of envelope 1 and 2 sequences of HCV, recovered from a subset of patients, we observed no apparent link between relatedness of patient sequences with culture viruses used and the corresponding neutralization results. In conclusion, pre-treatment levels of neutralizing antibodies against HCV genotype 1 isolates could not predict treatment outcome in patients with chronic HCV infection. High neutralization susceptibility of DH6/JFH1 could be correlated with adaptive envelope mutations previously highlighted as important for neutralization. Our study emphasizes the importance of using multiple culture viruses for neutralization studies and contributes to the current knowledge about neutralizing epitopes, important for future therapeutic- and vaccine-studies.

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The hepatitis C virus (HCV) envelope proteins E1 and E2 play a key role in host cell entry and represent important targets for vaccine and drug development. Here, we characterized HCV recombinants with chimeric E1/E2 complexes in vitro. Using genotype 1a/2a JFH1-based recombinants expressing 1a core-NS2, we exchanged E2 with functional isolate sequences of genotypes 1a (alternative isolate), 1b, and 2a. While the 1a-E2 exchange did not impact virus viability, the 2a-E2 recombinant was nonviable. After E2 exchange from three 1b isolates, long delays were observed before spread of infection. For recovered 1b-E2 recombinants, single E2 stem region amino acid changes were identified at residues 706, 707, and 710. In reverse genetic studies, these mutations increased infectivity titers by ~100-fold, apparently without influencing particle stability or cell binding although introducing slight decrease in particle density. In addition, the 1b-E2 exchange led to a decrease in secreted core protein of 25 to 50%, which was further reduced by the E2 stem region mutations. These findings indicated that compensatory mutations permitted robust infectious virus production, without increasing assembly/release. Studies of E1/E2 heterodimerization showed no differences in intracellular E1/E2 interaction for chimeric constructs with or without E2 stem region mutations. Interestingly, the E2 stem region mutations allowed efficient entry, which was verified in 1a-E1/1b-E2 HCV pseudoparticle assays. A CD81 inhibition assay indicated that the mutations influenced a late step of the HCV entry pathway. Overall, this study identified specific amino acids in the E2 stem region of importance for HCV entry and for production of infectious virus particles.

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A challenge for hepatitis C virus (HCV) vaccine development is defining conserved epitopes that induce protective antibodies against this highly diverse virus. An envelope glycoprotein (E2) segment located at amino acids (aa) 412 to 423 contains highly conserved neutralizing epitopes. While polyclonal antibodies to aa 412 to 423 from HCV-infected individuals confirmed broad neutralization, conflicting findings have been reported on polyclonal antibodies to an adjacent region, aa 434 to 446, that may or may not interfere with neutralization by antibodies to aa 412 to 423. To define the interplay between these antibodies, we isolated human monoclonal antibodies (HMAbs) to aa 412 to 423, designated HC33-related HMAbs (HC33 HMAbs), and characterized their interactions with other HMAbs to aa 434 to 446. A subset of the HC33 HMAbs neutralized genotype 1 to 6 infectious cell culture-derived HCV virions (HCVcc) with various activities. Although nonneutralizing HC33 HMAbs were isolated, they had lower binding affinities than neutralizing HC33 HMAbs. These antibodies could be converted to neutralizing antibodies by affinity maturation. Unidirectional competition for binding to E2 was observed between HC33 HMAbs and HMAbs to aa 434 to 446. When HMAbs to aa 434 to 446, which mediated neutralization, were combined with neutralizing HC33 HMAbs, biphasic patterns in neutralization were observed. A modest degree of antagonism was observed at lower concentrations, and a modest degree of synergism was observed at higher concentrations. However, the overall effect was additive neutralization. A similar pattern was observed when these antibodies were combined to block E2 binding to the HCV coreceptor, CD81. These findings demonstrate that both of these E2 regions participate in epitopes mediating virus neutralization and that the antibodies to aa 412 to 423 and aa 434 to 446 do not hinder their respective virus-neutralizing activities.

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Hepatitis C virus (HCV) is an important cause of chronic liver disease. Several highly diverse HCV genotypes exist with potential key functional differences. The HCV NS5A protein was associated with response to interferon (IFN)-α based therapy, and is a primary target of currently developed directly-acting antiviral compounds. NS5A is important for replication and virus production, but has not been studied for most HCV genotypes. We studied the function of NS5A using infectious NS5A genotype 1-7 cell culture systems, and through reverse genetics demonstrated a universal importance of the amphipathic alpha-helix, domain I and II and the low-complexity sequence (LCS) I for HCV replication; the replicon-enhancing LCSI mutation S225P attenuated all genotypes. Mutation of conserved prolines in LCSII led to minor reductions in virus production for the JFH1(genotype 2a) NS5A recombinant, but had greater effects on other isolates; replication was highly attenuated for ED43(4a) and QC69(7a) recombinants. Deletion of the conserved residues 414-428 in domain III reduced virus production for most recombinants but not JFH1(2a). Reduced virus production was linked to attenuated replication in all cases, but ED43(4a) and SA13(5a) also displayed impaired particle assembly. Compared to the original H77C(1a) NS5A recombinant, the changes in LCSII and domain III reduced the amounts of NS5A present. For H77C(1a) and TN(1a) NS5A recombinants, we observed a genetic linkage between NS5A and p7, since introduced changes in NS5A led to changes in p7 and vice versa. Finally, NS5A function depended on genotype-specific residues in domain I, as changing genotype 2a-specific residues to genotype 1a sequence and vice versa led to highly attenuated mutants. In conclusion, this study identified NS5A genetic elements essential for all major HCV genotypes in infectious cell culture systems. Genotype- or isolate- specific NS5A functional differences were identified, which will be important for understanding of HCV NS5A function and therapeutic targeting.

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Hepatitis C virus (HCV) is an important cause of chronic liver disease, and interferon-based therapy cures only 40 to 80% of patients, depending on HCV genotype. Research was accelerated by genotype 2a (strain JFH1) infectious cell culture systems. We previously developed viable JFH1-based recombinants encoding the structural proteins (core, E1, E2), p7, and NS2 of prototype isolates of the seven major HCV genotypes; most recombinants required adaptive mutations. To enable genotype-, subtype-, and isolate-specific studies, we developed efficient core-NS2 recombinants from additional genotype 1a (HC-TN and DH6), 1b (DH1 and DH5), and 3a (DBN) isolates, using previously identified adaptive mutations. Introduction of mutations from isolates of the same subtype either led to immediate efficient virus production or accelerated culture adaptation. The DH6 and DH5 recombinants without introduced mutations did not adapt to culture. Universal adaptive effects of mutations in NS3 (Q1247L, I1312V, K1398Q, R1408W, and Q1496L) and NS5A (V2418L) were investigated for JFH1-based genotype 1 to 5 core-NS2 recombinants; several mutations conferred adaptation to H77C (1a), J4 (1b), S52 (3a), and SA13 (5a) but not to ED43 (4a). The mutations permitting robust virus production in Huh7.5 cells had no apparent effect on viral replication but allowed efficient assembly of intracellular infectious HCV for adapted novel or previously developed recombinants. In conclusion, previously identified mutations permitted development of novel HCV core-NS2 genotype recombinants. Mutations adapting several recombinants to culture were identified, but no mutations were universally adaptive across genotypes. This work provides tools for analysis of HCV genotype specificity and may promote the understanding of genotype-specific patterns in HCV disease and control.

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We study the steady state of a phase-separated driven Ising lattice gas in three dimensions using computer simulations with Kawasaki dynamics. An external force field F(z) acts in the x direction parallel to the interface, creating a lateral order parameter current j^{x}(z) which varies with distance z from the interface. Above the roughening temperature, our data for "shearlike" linear variation of F(z) are in agreement with the picture wherein shear acts as effective confinement in this system, thus suppressing the interfacial capillary-wave fluctuations. We find sharper magnetization profiles and reduced interfacial width as compared to equilibrium. Pair correlations are more suppressed in the vorticity direction y than in the driving direction; the opposite holds for the structure factor. Lateral transport of capillary waves occurs for those forms of F(z) for which the current j^{x}(z) is an odd function of z , for example the shearlike drive, and a "steplike" driving field. For a V-shaped driving force no such motion occurs, but capillary waves are suppressed more strongly than for the shearlike drive. These findings are in agreement with our previous simulation studies in two dimensions. Near and below the (equilibrium) roughening temperature the effective-confinement picture ceases to work, but the lateral motion of the interface persists.

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We use a phase-separated driven two-dimensional Ising lattice gas to study fluid interfaces exposed to shear flow parallel to the interface. The interface is stabilized by two parallel walls with opposing surface fields, and a driving field parallel to the walls is applied which (i) either acts locally at the walls or (ii) varies linearly with distance across the strip. Using computer simulations with Kawasaki dynamics, we find that the system reaches a steady state in which the magnetization profile is the same as that in equilibrium, but with a rescaled length implying a reduction of the interfacial width. An analogous effect was recently observed in sheared phase-separated colloidal dispersions. Pair correlation functions along the interface decay more rapidly with distance under drive than in equilibrium and for cases of weak drive, can be rescaled to the equilibrium result.

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The effect of early nutrition on subsequent bone development was studied using gilts that had previously been fed ad libitum or 75% ad libitum intake and 100 or 150% National Research Council-recommended daily Ca and P from weaning to 100 kg. During the three-parity reproductive study, sows were fed a 14% protein diet. Metacarpals and metatarsals were taken from sows culled due to lameness or failure to breed and from sows after completing three parities. Femur and humerus articular cartilage and turbinates were described at necropsy. Metacarpals and metatarsals were heavier and tended to have thicker walls when sows were previously fed ad libitum or fed 150% Ca and P. Energy intake produced the greater response. Metacarpal breaking strength was greatest for sows previously fed ad libitum. Metatarsals were not affected by energy intake. Stiffness, Young's modulus of elasticity (YME) and flexural modulus for metacarpals and metatarsals were not affected by energy intake. Previously fed Ca and P intakes did not affect any of the mechanical bone characteristics. Metacarpals were heavier, had a greater breaking strength, were more elastic and exhibited slightly less resistance to bending than the metatarsals. The ether extract, ash, Ca and P content and the Ca:P ratio of metacarpals and metatarsals were not affected by previously fed energy or Ca and P intakes. The ether extract content tended to decrease and the ash, Ca and P content tended to increase with age. Articular cartilage and turbinate scores were not influenced by previously fed energy or Ca and P intakes. The YME and flexural modulus were the only bone characteristics that were even poorly correlated (average r = .25) with soundness scores, suggesting that high values result in poorer soundness scores. Energy and Ca and P intakes during growth and development had only minimal effects on bone characteristics and no apparent effect on structural soundness and longevity of sows kept for three parities.

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Gilts that had previously been fed ad libitum or 75% ad libitum intake and 100 or 150% National Research Council recommended daily Ca and P from weaning to 100 kg were used in a reproductive study in which a 14% protein diet was fed. Foot and leg measurements, subjective toe scores and structural soundness scores were taken at each of three parities, 21 d postweaning. Sows previously ad libitum-fed generally had larger front toes than limit-fed sows; whereas, hind toes were larger for sows previously fed 150% Ca and P levels than sows fed 100% Ca and P. Sows previously fed the ad libitum-150% Ca and P diet had the largest toes. Front inside toes were larger than hind inside toes, but the reverse was observed for front and hind outside toes, with the magnitude of the difference between the inside and outside toes greater for the hind foot. Toe size increased over parities with the greatest increase from parity 2 to 3. Incidence and severity of toe pad and horn scores were generally unaffected by previously fed energy and Ca and P levels and were not correlated to toe size. Hind feet exhibited a larger number of lesions than front feet and outside toes exhibited a larger number of lesions than inside toes, with the hind outside toe exhibiting the greatest number of lesions. In general, incidence and severity of toe lesions decreased or were unchanged from parity 1 to 3. Structural soundness scores were unaffected by previously fed energy or Ca and P levels, but were quadratically affected by parity, with a small increase (poorer) from parity 1 to 2 and a large improvement from parity 2 to 3. Soundness scores were not related to any of the feet or leg measurements and characteristics. Restricting growth rate and feeding elevated Ca and P levels during growth had no effect on incidence and severity of lesions on the toes and overall structural soundness of sows kept for three parities.

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Three groups of 96 crossbred gilts were weaned at 4 to 5 wk of age (average initial weight, 7.2 kg) and assigned to four dietary treatments in a 2 X 2 factorial arrangement of treatments (ad libitum vs 75% ad libitum and 100 vs 150% of NRC daily Ca and P). The effects of dietary treatments on the gait characteristics of gilts were analyzed by 16-mm motion picture photography. At approximately 50 and 100 kg body weight (periods 1 and 2, respectively), pigs were photographed walking on a treadmill, and gait characteristics were measured from the motion picture film. Ad libitum-fed gilts were longer, taller, wider and deeper when compared with restricted-fed gilts on an equal age basis, but not when compared on an equal weight basis. Other side view and rear view measurements were inconsistently affected by energy level (ad libitum vs restricted) at the two time periods, and all measurements were unaffected by Ca and P level. Analysis of the side and rear view characteristics over time generally revealed undesirable changes (P less than .01) from 59 to 100 kg. Both left and right hock-joint deviation increased (P less than .01) from period 1 to period 2, when expressed on an equal weight basis, suggesting the development of joint weakness. The hind pastern angle and the angle at the hock-joint also increased with time (P less than .01), suggesting development of post-leggedness as pigs increased in age and weight. Correlation coefficients among the various photographic characteristics and structural soundness scores were generally very low. Long-term effects of the dietary treatments on structural development are under study.

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Forty-four purebred Yorkshire boars, reared outdoors on concrete, were randomly assigned to one of three age groups (150 +/- 7, 200 +/- or 250 +/- 7 days of age) for the purpose of examining endogenous testosterone concentration in response to one of four exogenously administered treatments. Four boars from each age group were administered either human chorionic gonadotrophin (hCG; 1000 U.S.P. units intravenous), adrenocorticotrophin (ACTH; 100 IU intravenous), or testosterone proprionate (TP; 25 mg intramuscular). The remaining boars served as controls and were given saline (S; 5 ml IV). Blood samples were collected from each boar at -120, -90, -60, -30, and 0 min pre-treatment and at 15, 30, 45, 60, 75, 90, 105, 120, 150, 180, 210, 240, 270, 300, 330, and 360 min post-treatment via an indwelling anterior vena cava catheter. Plasma testosterone was quantified by radioimmunoassay. Within treatment, boars receiving hCG, ACTH or S responded similarly (P>.10) across the three age groups for measured testosterone. Plasma testosterone was elevated (P<.05) by 30 min (4.7 +/- .5 ng/ml; X +/- SEM) and 15 min (5.5 +/- 1.1 ng/ml) post-treatment in boars administered hCG and ACTH, respectively, when compared with the S (1.0 +/- .3 ng/ml) group. Testosterone in hCG treated boars peaked by 90 min (21.8 +/- 1.8 ng/ml) post-treatment, declined slightly until 210 min (18.8 +/- 1.8 ng/ml) post-treatment and increased thereafter. Hormone levels in ACTH treated boars plateaued by 45 min (7.5 +/- 1.4 ng/ml) post-treatment and began to decline by 90 min post-treatment. Plasma testosterone for TP treated boars differed (P<.05) over time among the three age groups. Boars 150 +/- 7 and 200 +/- 7 days of age had an increase in plasma testosterone at 150 and 240 min post-treatment with TP, respectively. Results suggest that the testosterone biosynthetic and secretory capabilities of the boar testes are fully operational by 150 days of age.