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Transferases are ubiquitous across all known life. While much work has been done to understand and describe these essential enzymes, there have been minimal efforts to exert tight and reversible control over their activity for various biotechnological applications. Here, we apply a rational, computation-guided methodology to design and test a transferase-class enzyme allosterically regulated by light-oxygen-voltage 2 sensing domain. We utilize computational techniques to determine the intrinsic allosteric networks within N-acyltransferase (Orf11/∗Dbv8) and identify potential allosteric sites on the protein’s surface. We insert light-oxygen-voltage 2 sensing domain at the predicted allosteric site, exerting reversible control over enzymatic activity. We demonstrate blue-light regulation of N-acyltransferase (Orf11/∗Dbv8) function. Our study for the first time demonstrates optogenetic regulation of a transferase-class enzyme as a proof-of-concept for controllable transferase design. This successful design opens the door for many future applications in metabolic engineering and cellular programming.

Summary Six broodmares that were genetically tested to be heterozygous (H/N) and six broodmares that were tested to be homozygous negative (N/N) for Equine Hyperkalemic Periodic Paralysis (HYPP), that were descendents of the same stallion, were used in a replicated 3×3 Latin square experiment to determine the genetic-diet relationships in the HYPP syndrome.1 The mares were fed rations consisting of 65% pelleted concentrate and 35% Coastal Bermuda grass hay that provided 1.1 (diet A), 1.9 (diet B) and 2.9% (diet C) potassium. The experimental periods were 14 days long, resulting in 28 meals per period. At meals 1 and 27, blood samples were taken every 30 minutes for 12 hours and every 10 minutes from 2 to 5 hours post-feeding. Total urine and fecal collections were made on the last 4 days of each period. Water, feed and fecal samples were analyzed for sodium and potassium content. Blood was analyzed for packed cell volume (PCV) and plasma potassium (K+) and sodium (Na+) concentrations. Apparent absorption of potassium was 99.8% across all diets, and potassium was excreted principally in the urine. More potassium was retained when the horses were fed diet B than diets A or C. Apparent absorption of sodium was 99.6% with no difference by diet. Neither potassium nor sodium balances were affected by HYPP status. There was no post-prandial pattern of plasma K+ concentration seen when the horses were fed the low potassium diet. When fed the higher potassium diets, they had greater plasma K+ values and exhibited a post-prandial peak at 2 to 5 hours after feeding. This peak occurred in all of the horses, meaning that plasma K+ cannot be used as a diagnostic aid for the presence of the HYPP mutation. By meal 27, some adaptation had occurred resulting in lower plasma K+ values when the horses were fed the higher potassium diets. The horses had increased plasma K+ concentrations throughout the post-prandial period while fed diet B compared to diet A, agreeing with the positive potassium balance seen in this experiment. The HYPP H/N horses had lower plasma Na+ concentrations in the early post-prandial hours and continuously greater PCV values than the HYPP N/N horses.

Summary During an experiment to determine the genetic-diet interactions in genetically predisposed Quarter Horses, biphasic symptoms of Hyperkalemic Periodic Paralysis (HYPP) were observed when the HYPP H/N broodmares were fed the higher potassium rations. 1 The cause of those symptoms was assumed to be: 1) the difference in absorption times of the highly absorbable potassium citrate in the concentrates and the potassium contained in the cells of the grains and hays in the rest of the rations, 2) partial absorption of potassium in the small intestine followed by no absorption from the cecum and further absorption in the large colon or 3) a combination of both of the above explanations. Since previously cannulated ponies were available in this laboratory, an experiment was designed to determine which of those possibilities was the most likely. Three ponies, weighing an average of 132 kg, with posterior-ileal cannulas were used in a 3×3 Latin square experiment to determine the amounts of potassium and sodium that would be absorbed pre-cecally and post-ilealy. The ponies were fed the same rations as the mares in the previously described experiment (see part I), which provided approximately 1.1, 1.9 and 2.9% potassium by weight as fed. Samples of water, feces, ileal contents and blood were collected. Water, feed, ileal and fecal samples were analyzed for sodium and potassium concentration. Apparent absorption of potassium prior to the cecum was 97% when the ponies were fed the low potassium diet and 98% when they were fed the higher potassium diets. Three percent and 2% of the potassium was absorbed from the hindgut when the ponies were fed the low potassium diet and the higher potassium diets, respectively. Nine percent and 16% of sodium was absorbed pre-cecally when the ponies were fed the low potassium diet and the higher potassium diets, respectively. Ninety-one and 84% of the sodium was absorbed from the hindgut, when the ponies were fed the low potassium diet and the higher potassium diets, respectively for 99–100% total absorption of both sodium and potassium from all diets. These results indicate that the potassium from the three diets was absorbed pre-cecally, and that the process took up to nine hours, resulting in bi-phasic HYPP symptoms in the HYPP H/N mares in the companion experiment. This information can be used to better understand the relationship between dietary potassium, plasma K + concentration and HYPP symptoms.

Summary Six broodmares that were DNA tested to be heterozygous (H/N) and six broodmares that were tested to be homozygous negative (N/N) for Equine Hyperkalemic periodic Paralysis (HYPP), that were descendents of the same stallion, were used in a replicated 3x3 Latin square experiment to determine the genetic-diet relationships in the HYPP syndrome. 1 The mares were fed rations consisting of 65% pelleted concentrate and 35% Coastal Bermuda grass hay that provided 1.1 (diet A), 1.9 (diet B) and 2.9% (diet C) potassium in two evenly spaced meals per day. The experimental periods were 14 days long, resulting in 28 meals per period. The HYPP Symptoms Index was greater at meal 1 than meal 27 and greater when the horses were fed diet C than diet B. Plasma K + concentrations at the onset of symptoms were not different by diet or meal and averaged 3.78 mmol/L. The average time of onset of symptoms was two hours and 33 minutes after the beginning of a meal, which was not different by diet or meal. A regression analysis of 204 paired non-zero HYPP Symptoms Index values and their corresponding plasma K + concentrations at the same times, from the HYPP H/N horses, resulted in an R 2 of .449. While fed diet A, the horses' mean plasma K + values remained below 3.5 mmol/L, and no symptoms were observed. Generally, an increase in Symptoms Index followed increased plasma K + concentration when the horses were fed diets B and C at meal 1. At meal 27, the mean HYPP Symptoms Index and average plasma K + values were lower than at meal 1. However, there were increased plasma K + concentrations observed in the later post-prandial hours after meal 27, when the horses were fed diet B, that did not correspond to HYPP symptoms. In a pilot study, one horse collapsed into sternal recumbency after meal 1 when her HYPP Symptoms Index was 10 and her plasma K + concentration was 7.29 mmol/L. This horse spontaneously recovered from the episode. By meal 27, both the peak plasma K + concentration (4.6 mmol/L) and the peak HYPP Symptoms Index (1) were smaller in this horse, but they still occurred simultaneously.

Summary Six broodmares that were genetically tested to be heterozygous (H/N) and six broodmares that were tested to be homozygous negative (N/N) for Equine Hyperkalemic Periodic Paralysis (HYPP), that were descendents of the same stallion, were used in a replicated 3x3 Latin square experiment to determine the genetic-diet relationships in the HYPP syndrome. The mares were fed rations consisting of 65% pelleted concentrate and 35% Coastal Bermudagrass hay that provided 1.1 (diet A), 1.9 (diet B) and 2.9% (diet C) potassium in the total rations. Symptoms of HYPP, such as repeated yawning, muscle twitching, generalized muscle fasciculations, myotonia, irregular movement, prolapse of the membrana nicitans and involuntary recumbency, were recorded during each 14-day experimental period. There were no HYPP symptoms in the HYPP N/N horses fed any diet, or in the HYPP H/N horses fed the low potassium diet. Symptoms were observed after 52% of the horse-meal combinations when the HYPP H/N horses were fed the medium potassium diet and after 67% of the horse-meal combinations when they were fed the high potassium diet. The onset and longest duration of symptoms were on the head or shoulders 66% of the time. Bi-phasic symptoms were rare and were more frequent after diet C than diet B. Symptoms were twice as common in the day as the night. These results indicate that a genetic-diet interaction exists in the HYPP syndrome and that dietary control of HYPP symptoms is possible in genetically susceptible horses.

Summary Fifty-three Quarter Horses were put into race-training at 18 mo of age. Changes in the third metacarpal were monitored by radiographic densitometry initially at 83 days prior to the commencement of training and at days 0, 62, 104 and 244 of training. A normal increase in density of the third metacarpal due to growth and mineralization was seen from the first set of radiographs until the horses began training at day 0 (P

Summary Beginning at approximately 18 mo of age, fifty-three Quarter Horses were trained to ride, conditioned and run in nine match races with three each at 274 m, 320 m and 366 m. As part of a companion study, the horses were fed varying amounts of Sodium Zeolite A. During the last long slow distance (LSD) workout, the last LSD with sprints workout prior to racing, and each race, blood samples were collected and heart rates (HR), respiration rates (RR) and rectal temperatures (RT) were measured at rest, after the warm-up, and at 1, 2, 5, 10, 20, 30, 60 and 120 min of recovery. Blood samples were analyzed for packed cell volume (PCV) and plasma ionized calcium (Ca++), sodium (Na+), potassium (K+), lactate (LA − ) and glucose (GLU) concentrations. Data were analyzed by ANOVA using the general linear model of SAS appropriate for repeated measures and Scheffe's multiple comparison test to compare diet, race and sample (time) effects. Results were considered significant at P The horses were able to run increasingly longer races without corresponding changes in HR, PCV or plasma Na+, K+, Ca++, LA - or GLU concentrations. The warm-up elicited an average HR of 163 beats/min, yet HR during and immediately after the races did not go as high as commonly observed in Thoroughbred racehorses running longer distances. Respiration rate and RT were higher during recovery from the longer races in response to increased metabolically generated heat.

Summary Sixty weanlings were maintained in dry lots without forced exercise and individually fed a diet consisting of 25% Bermudagrass hay and 75% balanced concentrate twice dally. Radiographs of the carpus, metacarpus and first phalanx, and other radiographs of the tarsal region, were taken initially at approximately 180 d of age and at approximately 236, 292 and 348 d of age. The radiographs were evaluated for the presence or absence of subchondral and/or cystic lesions in the carpal and tarsal regions and for closure of physes of the distal third metacarpus and proximal first phalanx. Physeal scores were: 1 = open; 2 -- partially closed; 3 = closed. Only 1.7% of the horses had subehondral lesions in the carpal region at 180 d, and none had those lesions at 236, 292 or 348 d. There was a 3.3% incidence of subchondral lesions of the tarsus at 180, 236 and 292 d, but only 1.7% at 348 d. Cystic lesions of the carpus were evident in 13.3% of the horses at 180 d, 16.7% at 236 d and 18.3% at 292 and 348 d. None of the hrses had cystic lesions oftbe tarsus at 180, 236 or 348 d, but 1.7% had them at 292 d. Physeal closure scores for the distal metacarpus averaged 1.5 and 2.6 at 180 and 236 d, respectively. At 292 and 348 d, all of the distal metacarpi were closed. Physeal closure scores for the proximal first phalanges were 1.5, 2.6 and 2.9 at 180

Procedure Number HNF-PRO-408, revision 0, paragraph 1.0, ''Purpose,'' and paragraph 2.0, ''Requirements for Facility Management of Asbestos,'' relate building inspection and requirements for documentation of existing asbestos-containing building material (ACBM) per each building assessment. This documentation shall be available to all personnel (including contractor personnel) entering the facility at their request. Corrective action was required by 400 Area Integrated Annual Appraisal/Audit for Fiscal Year 1992 (IAA-92-0007) to provide this notification documentation. No formal method had been developed to communicate the location and nature of ACBM to maintenance personnel in the Fast Flux Test Facility (FFTF) 400 Area. The scope of this Data Package Document is to locate and evaluate any ACBM found at FFTF which constitutes a baseline. This includes all buildings within the protected area. These findings are compiled from earlier reports, numerous work packages and engineering evaluations of employee findings.

University, Ohio University and University of New Hampshire (Level 3). In2011, a total of 11 studentsbecamecertified RegisteredLevel Therapeutic Riding Instructors through this program. In 2012, the number of university students certified through this new program was 14.Higher Education Membership growth is occurring; however, with the addition of 3 new universities joining for 2013. It is important to point out that the Instructor certification within Higher Education remain small to date (2.3 and 3.7% of the total PATH Intl. Instructor certifications in 2011 and 2012, respectively), with the majority of certifications associated with private training centers and workshop/ certifications at private centers. However, with the growing implementation of equine-assisted therapy curriculum in over 30 colleges, junior colleges and universities in the United States, the future of the equine assisted therapy industry may see a shift in training of young college-aged individuals from private centers and training programs to those within Higher Education. Further, although all certifications to date through Higher Education members are at the Registered level, it is also foreseen that in the near future, more students will need to be certified at the Advanced Level to meet the growing need of the Industry for Advanced Instructors in position so Program Directors, Head Instructors and Executive Directors of Centers in the Field of Equine Assisted Therapy.

Osteoarthritis (OA) and Osteochondritis Dissecans (OCD) are osteoarticular disorders that cause leg weakness, lameness, pain and suffering in companion animals, some farm animals and humans. OA is one of the most common age-related osteoarticular disorders in humans and dogs. In pigs, both OA and OCD are thought to arise from changes in the articular cartilage and growth plates within the synovial joints causing structural damage to joint tissues. Since these changes are not observed in the slow maturing wild boar, they are suggested to be a result of the modern intensive pig production industry which has very successfully selected pigs for rapid growth rates, large muscle mass and efficient feed conversion placing increased weight and mechanical stress on growth plates. The aim of this study was to establish canine and porcine articular cartilage explant models which are essentially tissue culture techniques for isolating and maintaining cartilage tissue ex vivo for subsequent assessment of potentially beneficial effects of specific phytonutrients. Bacterial lipopolysaccharide (LPS) was used as a catabolic mediator to create a culture model of joint inflammation mimicking the events that occur in late stages of OA and OCD. We then performed assays to determine if the dietary phytochemical ‘curcumin’ (derived from Curcuma longa) and the polyphenolic phytoalexin stilbene ‘resveratrol’ (found in red grapes, red wine, peanuts and some berries) are able to counteract the catabolic effects of LPS by inhibiting LPS stimulated release of cartilage matrix glycosaminoglycans (GAGs).

The importance is shown of energy saving as a factor determining the future of the cast-metals industry, which is currently in decline in most of the advanced industrialised countries of the world, except perhaps in Japan. The paper highlights the advantages and disadvantages of the casting process in an energy-saving context, and comes to some conclusions about future prospects.

Rift Valley fever virus (RVFV), a member of the family Bunyaviridae, extended its range from sub-Saharan Africa into Egypt in 1977. Its clinical spectrum is recognized to include severe manifestations such as hemorrhagic fever and encephalitis. For these reasons, as well as the limited knowledge of specific therapy for Bunyaviridae infections, we investigated several prophylactic regimens for RVF in a mouse model. Rimantadine, thiosemicarbazone, and inosiplex were ineffective. Pretreatment with glucan was of some use, but the most encouraging results were obtained with the antiviral drug ribavirin, passive antibody, or an interferon inducer polyriboinosinic-polyribocytidylic acid complexed with poly-L-lysine and carboxymethylcellulose (poly[ICLC]). Ribavirin and poly(ICLC) were also shown to be efficacious in preventing disease in hamsters. Ribavirin (loading dose of 50 mg/kg followed by 10 mg/kg at 8-h intervals for 9 days) suppressed viremia in RVF-infected rhesus monkeys. Ribavirin also reduced virus yield in infected cell cultures; sensitivity varied markedly with cell type but not with virus strain. Immune mouse ascitic fluid, with a plaque reduction neutralization titer of 1:1024, was effective in a dose of 4 ml/kg, a volume approximately equivalent to administration of a unit of convalescent plasma to a human. Poly(ICLC) may well have functioned through interferon induction, since RVFV was shown to be sensitive to interferon in cell culture, and since another macrophage activator (glucan) was only marginally effective. These studies suggest that ribavirin, poly(ICLC), and convalescent plasma may have a role in prevention or therapy of human RVF.

A great variety of surface coatings and treatments is used to combat wear and corrosion in the many types of rotary and reciprocating engines now in service. This paper is primarily concerned with the tribological behaviour of engine components, but corrosion and other problems are referred to where the different elements of functional performance are inseparable. The paper provides a brief review of current surface treatment and indicates the processes chosen for different engine components, indicating the reasons for the choice. Recently developed surface treatments are surveyed and their potential, for future engineering application, is assessed.

This paper sets out to give a balanced and comprehensive account of the progress made in the ‘casting industry’. Casting is fundamental to the metals industry and to the national economy as a whole. UK business in the field probably exceeds £700m p.a. and provides work for over 150 000 employees, as well as underpinning much of the engineering industry.At present different branches of the industry are facing an increasing number of common problems, and finding much common ground in their response to them.