Standardized ileal digestible amino acids and digestible energy contents in two modified soy protein concentrates and soybean meal fed to growing pigs.

Six ileal-cannulated barrows (28.0 ± 1.3 kg initial BW) were used in a replicated 3 × 3 Latin square design with one additional period (n = 7 or 6) to determine standardized ileal digestible (SID) AA and digestible energy of two modified soy protein concentrates [MSPC1 and MSPC2] and soybean meal (SBM). Pigs were fed one of three cornstarch-based diets with either MSPC1 or MSPC2 or SBM as the sole source of AA at a rate of 2.8 times the estimated maintenance energy requirement. In each period, pigs were adapted to diets for 7 d followed by 2 d of fecal collection and subsequently, 2 d of continuous ileal digesta collection for 8 h. The SID of AA was calculated using basal endogenous losses from a previous study for pigs fed a nitrogen-free diet. The digestible energy of the ingredients was calculated according to the difference method using a nitrogen-free diet that contained the same cornstarch:sucrose:oil ratio as the three test diets. The total Lys content was 33% and 38% greater for MSPC1 vs. MSPC2 and SBM, respectively. The SID of crude protein was greater for MSPC1 (96.9%) than for SBM (91.3%; P < 0.05), whereas an intermediate value was observed for MSPC2 (94.3% ± 1.2%). The SID of Ile (93.8%), Leu (93.6%), Lys (93.9%), Phe (96.7%), and Val (93.2%) were not different between MSPC1 and MSPC2 but greater than for SBM (88.8% ± 1.3%, 87.8% ± 1.2%, 84.5% ± 1.7%, 92.9% ± 1.0%, 86.5% ± 1.7% for Ile, Leu, Lys, Phe, and Val, respectively; P < 0.05). The SID of His and Thr was greater for MSPC1 than MSPC2 and SBM (P < 0.05), which were not different. The SID of Met was greater for MSPC1 and SBM vs. MSPC2 (P < 0.05). The SID of Arg was greater for MSPC1 than MSPC2 and SBM (P < 0.05), and greater for MSPC2 than SBM (P < 0.05). The digestible energy was greater for MSPC1 (4,677 kcal/kg) than MSPC2 and SBM (average; 3,896 ± 239 kcal/kg; P < 0.05), which were not different. Therefore, the MSPC1 was a better source of SID Lys and digestible energy than either MSPC2 or SBM and could be used as a highly digestible protein ingredient in swine rations.

Expression of Cathepsins B, D, and G in Extracranial Arterio-Venous Malformation.

Objectives: We have previously identified a population of cells that expressed stemness-associated markers in extracranial arterio-venous malformation (AVM) and demonstrated expression of cathepsins B, D, and G on embryonic stem cell (ESC)-like populations in other vascular anomalies. This study investigated the expression of cathepsins B, D, and G, and their localization in relation to this primitive population in extracranial AVM. Methods: Immunohistochemical staining was performed on AVM tissue samples from 13 patients to demonstrate expression of cathepsins B, D, and G. Western blotting was performed on four AVM tissue samples and three AVM-derived primary cell lines to confirm protein expression of cathepsins B and D proteins. RT-qPCR was performed on three AVM-derived primary cell lines to demonstrate transcript expression of cathepsins B, D, and G. Enzymatic activity assays were performed on three AVM-derived primary cell lines to investigate if cathepsins B and D were active. Localization of the cathepsins was investigated using immunofluorescence dual-staining of the cathepsins with the ESC markers OCT4 and SOX2, and mast cells marker chymase on two of the 13 AVM tissue samples. Results: Immunohistochemical staining demonstrated expression of cathepsins B, D, and G in all 13 AVM tissue samples. Western blotting showed expression of cathepsins B and D proteins in all four AVM tissue samples and all three AVM-derived primary cell lines. RT-qPCR demonstrated transcripts of cathepsins B, D, and G in all three AVM-derived primary cell lines. Enzymatic activity assays showed that cathepsins B and D were active. Immunofluorescence staining showed expression of cathepsins B and D on the OCT4+/SOX2+ endothelium and media of the lesional vessels and cells within the stroma in AVM nidus. Cathepsin G was expressed on the chymase+ phenotypic mast cells. Conclusions: This study demonstrated the novel finding of the expression of cathepsins B, D, and G in AVM. Cathepsins B and D were expressed by the primitive population, and cathepsin G was localized to mast cells, within the AVM nidus.

The effects of precisely meeting estimated daily energy and lysine requirements for gestating sows over three consecutive pregnancies on sow reproductive and lactation performance.

The objective of the current study was to determine the effects of precisely meeting estimated daily energy and Lys requirements for gestating sows over three consecutive pregnancies on sow reproductive and lactation performance. A total of 105 sows (initial reproductive cycle 1.4 ± 0.5) were randomly assigned to a precision (PF; n = 50) or control (CON; n = 55) feeding program between days 2 and 9 of gestation and housed in group-pens equipped with electronic sow feeders capable of blending two diets. The PF sows received unique daily blends of two isocaloric diets (2518 kcal/kg NE; 0.80% and 0.20% standardized ileal digestible [SID] Lys, respectively), whereas CON sows received a static blend throughout gestation to achieve 0.56% SID Lys. After weaning, sows were re-bred and entered the same feeding program as in the previous pregnancy for two subsequent pregnancy cycles (PF: n = 36; CON: n = 37; average reproductive cycle: 2.4 ± 0.5; PF: n = 25; CON: n = 24; average reproductive cycle: 3.5 ± 0.5). Sows on the PF program received 97%, 105%, and 118% (average over three pregnancy cycles) of dietary energy and 67%, 79%, and 106% of SID Lys intakes compared to CON between days 5 and 37, 38 and 72, and 73 and 108 of gestation, respectively. Estimated N (26.1%) retention did not differ between gestation feeding programs in any pregnancy, but excess N excretion was less (1617 vs. 1750 ± 54 g/sow; P < 0.01) for PF vs. CON sows. Regardless of pregnancy cycle, sows that received the PF program had greater ADG between days 38 and 72 (614 vs. 518 ± 63 g/d; P < 0.05) and between days 73 and 108 (719 vs. 618 ± 94 g/d; P = 0.063) of gestation, and greater loin depth gain between days 63 and 110 of gestation (0.7 vs. -1.1 ± 1.6 mm; P < 0.05), but BW (235.1 kg) and backfat (17.8 mm) and loin (70.5 mm) depths on day 110 of gestation did not differ. The number of piglets born alive, stillborn, and mummified, and litter birth weight (16.5 kg) did not differ in any pregnancy cycle, nor did piglet ADG during lactation (250 g/d) and piglet BW (6.7 kg) at weaning. Sows that received the PF program during gestation had lower ADFI during lactation (5.7 vs. 6.2 ± 0.2 kg; P < 0.01). Therefore, using feeding programs that precisely match estimated daily energy and Lys requirements for gestating sows provides the opportunity to reduce N losses to the environment and reduce lactation feed usage, without negatively affecting sow reproductive and lactation performance.

Precision feeding gestating sows: effects on offspring growth performance and carcass and loin quality at slaughter.

A total of 601 pigs from 65 litters were used to determine the effects of closely meeting estimated daily Lys and energy requirements for sows during gestation for three consecutive parities on offspring postweaning growth performance and carcass and loin quality at slaughter. Sows were assigned a control (static diet composition; CON) or precision (individual daily blend of two diets to meet estimated Lys and energy requirements; PRE) feeding program between days 7 and 110 of gestation for three consecutive pregnancy cycles, starting with primiparous sows (parity 1: 12 CON and 12 PRE sows; parity 2: 8 CON and 13 PRE sows; parity 3: 8 CON and 12 PRE sows). At weaning (20 ± 2 d of age), up to 10 pigs per litter were randomly selected and placed in a pen (1 litter per pen). All pens received ad libitum access to commercial diets in six phases (four-phase nursery, grower, and finisher, respectively). Four pigs per pen were slaughtered at ~125 kg BW for evaluation of carcass characteristics and loin quality. The ADG and ADFI of offspring were not influenced by maternal feeding program in any parity during nursery phases I through III. During nursery phase IV, ADG and ADFI were greater for litters from PRE- vs. CON-fed sows (0.70 vs. 0.66 ± 0.03 and 1.15 vs. 1.08 ± 0.06 kg/d for ADG and ADFI, respectively; P < 0.05). The BW for litters from PRE- vs. CON-fed sows tended to be greater by day 66 of age (end of nursery period; 29.7 vs. 28.7 ± 1.1 kg; P = 0.076). Within the grower phase, litters from PRE-fed sows had a greater ADG in parity 2 but lower ADG in parity 3 vs. litters from CON-fed sows (0.99 vs. 0.94 and 0.93 vs. 1.01 ± 0.03 kg/d for parities 2 and 3, respectively; P < 0.05). No differences were observed for ADG or ADFI in the finisher phase or G:F in any phase for any parity. Loin eye area was smaller (52.2 vs. 55.0 ± 1.8 cm2; P < 0.05) for offspring from PRE- vs. CON-fed sows. In parity 2, carcass lean yield tended to be less for offspring from PRE- vs. CON-fed sows (58.6 vs. 59.6 ± 0.4%; P = 0.051). Minimal differences were observed for subjective and objective evaluations of loin quality. Closely meeting the estimated daily energy and Lys requirements for sows throughout gestation for three consecutive pregnancy cycles improved offspring growth performance (ADG and ADFI) in the final nursery stage, but generally did not affect growth performance in grower/finisher periods or carcass and loin quality at ~125 kg BW.

Cell Populations Expressing Stemness-Associated Markers in Vascular Anomalies.

Treatment of vascular anomalies (VAs) is mostly empirical and, in many instances unsatisfactory, as the pathogeneses of these heterogeneous conditions remain largely unknown. There is emerging evidence of the presence of cell populations expressing stemness-associated markers within many types of vascular tumors and vascular malformations. The presence of these populations in VAs is supported, in part, by the observed clinical effect of the mTOR inhibitor, sirolimus, that regulates differentiation of embryonic stem cells (ESCs). The discovery of the central role of the renin-angiotensin system (RAS) in regulating stem cells in infantile hemangioma (IH) provides a plausible explanation for its spontaneous and accelerated involution induced by ß-blockers and ACE inhibitors. Recent work on targeting IH stem cells by inhibiting the transcription factor SOX18 using the stereoisomer R(+) propranolol, independent of ß-adrenergic blockade, opens up exciting opportunities for novel treatment of IH without the ß-adrenergic blockade-related side effects. Gene mutations have been identified in several VAs, involving mainly the PI3K/AKT/mTOR and/or the Ras/RAF/MEK/ERK pathways. Existing cancer therapies that target these pathways engenders the exciting possibility of repurposing these agents for challenging VAs, with early results demonstrating clinical efficacy. However, there are several shortcomings with this approach, including the treatment cost, side effects, emergence of treatment resistance and unknown long-term effects in young patients. The presence of populations expressing stemness-associated markers, including transcription factors involved in the generation of induced pluripotent stem cells (iPSCs), in different types of VAs, suggests the possible role of stem cell pathways in their pathogenesis. Components of the RAS are expressed by cell populations expressing stemness-associated markers in different types of VAs. The gene mutations affecting the PI3K/AKT/mTOR and/or the Ras/RAF/MEK/ERK pathways interact with different components of the RAS, which may influence cell populations expressing stemness-associated markers within VAs. The potential of targeting these populations by manipulating the RAS using repurposed, low-cost and commonly available oral medications, warrants further investigation. This review presents the accumulating evidence demonstrating the presence of stemness-associated markers in VAs, their expression of the RAS, and their interaction with gene mutations affecting the PI3K/AKT/mTOR and/or the Ras/RAF/MEK/ERK pathways, in the pathogenesis of VAs.

Total Joint Arthroplasty in Patients Taking Methadone or Buprenorphine/Naloxone Preoperatively for Prior Heroin Addiction: A Prospective Matched Cohort Study.

BACKGROUND: Preoperative narcotic use has been associated with poor outcomes after total joint arthroplasty (TJA). The purpose of this study is to compare clinical outcomes of patients undergoing elective TJA while concurrently being treated with methadone or buprenorphine/naloxone for prior heroin addiction to a matched control group. METHODS: From an electronic medical record, we collected age, gender, body mass index, the presence of back pain, smoking status, history of alcohol abuse, preoperative use of a pain clinic, and use of antipsychotics, antidepressants, or systemic corticosteroids. Validated outcome measures including the 12-Item Short Form Survey, Knee Society Score (KSS), and Harris Hip Score were used to assess functional outcomes preoperatively and postoperatively. Perioperative data were retrospectively obtained from patient charts. Postoperative functional outcomes were prospectively collected at follow-up visits. Subjects were matched to 2:1 control group on the basis of procedure, sex, diagnosis, age (±5 years), and body mass index (±5 kg/m(2)). Average follow-up was 27.2 months. RESULTS: Significant preoperative differences between the groups included mean morphine-equivalent requirements (997.1 mg for study group vs 24.8 mg for controls), 12-Item Short Form Survey Mental Component Scores (MCS-12; 37.8 for study group vs 49.0 for controls), smoking history, and antipsychotic medication use. Perioperative referral to inpatient Acute Pain Service and mean in-hospital morphine-equivalent narcotic usage (793 mg/24 h for study group vs 109 mg/24 h for controls) also significantly differed between groups. Knee range of motion differed significantly between the cohorts at 1 year (77.5 for study group vs 109.4); however, no significant difference in KSS pain (87.6 vs 84.4), KSS function (61 vs 80.9), Harris Hip Score (89.2 vs 85.3), MCS-12 (47.1 vs 52.9), or complications was observed. CONCLUSION: Equivalent pain control and successful clinical outcome at 1 year can be achieved in patients who use methadone or buprenorphine/naloxone preoperatively.

Laryngeal melanosis.

OBJECTIVE: Laryngeal melanosis (LM) is an unusual condition. Little is known about its clinical significance. To learn more about this unusual disorder, we prospectively gathered data on a series of patients with LM and compared the data with the summarized findings from the cases currently available in the literature. STUDY DESIGN: Case series with planned data collection. SETTING: Two hundred fifty-five-bed urban indigent care hospital located on a university medical center campus. Subjects and Methods Demographics, history, examination findings, and concurrent head and neck disorders were prospectively recorded for all patients with LM who presented to our institution between 2005 and 2012. RESULTS: Eighteen patients with laryngeal melanosis were enrolled. All patients are African American, and all have a history of chronic tobacco use. The most common presenting symptom was dysphonia, and examination most often revealed flat, pigmented lesions of the larynx. Nine patients (50%) had concomitant squamous cell carcinoma of the upper aerodigestive tract. There are 19 previous cases of laryngeal melanosis reported by 12 different authors, with a concomitant carcinoma rate of 26%. Our patients are similar in age and are all African American, with a greater percentage of women and greater rate of concomitant carcinoma. Our study represents the single largest case series of laryngeal melanosis and is more complete given the prospective nature of the data collection. CONCLUSION: Laryngeal melanosis is an unusual disorder that is associated with chronic tobacco use. Clinicians should be aware of the association between laryngeal melanosis and squamous cell carcinoma and perform a thorough evaluation for concomitant carcinoma.

Dolphin underwater bait-balling behaviors in relation to group and prey ball sizes.

We characterized dusky dolphin (Lagenorhynchus obscurus) feeding behaviors recorded on underwater video, and related behaviors to variation in prey ball sizes, dolphin group sizes, and study site (Argentina versus New Zealand, NZ). Herding behaviors most often involved dolphins swimming around the side or under prey balls, but dolphins in Argentina more often swam under prey balls (48% of passes) than did dolphins in NZ (34% of passes). This result may have been due to differences in group sizes between sites, since groups are larger in Argentina. Additionally, in NZ, group size was positively correlated with proportion of passes that occurred under prey balls (p<0.001). Prey-capture attempts most often involved capturing fish from the side of prey balls, but dolphins in Argentina more often swam through prey balls (8% of attempts) than did dolphins in NZ (4% of attempts). This result may have been due to differences in prey ball sizes between sites, since dolphins fed on larger prey balls in Argentina (>74m(2)) than in NZ (maximum 33m(2)). Additionally, in NZ, dolphins were more likely to swim through prey balls to capture fish when they fed on larger prey balls (p=0.025).

Specification of motoneurons from human embryonic stem cells.

An understanding of how mammalian stem cells produce specific neuronal subtypes remains elusive. Here we show that human embryonic stem cells generated early neuroectodermal cells, which organized into rosettes and expressed Pax6 but not Sox1, and then late neuroectodermal cells, which formed neural tube-like structures and expressed both Pax6 and Sox1. Only the early, but not the late, neuroectodermal cells were efficiently posteriorized by retinoic acid and, in the presence of sonic hedgehog, differentiated into spinal motoneurons. The in vitro-generated motoneurons expressed HB9, HoxC8, choline acetyltransferase and vesicular acetylcholine transporter, induced clustering of acetylcholine receptors in myotubes, and were electrophysiologically active. These findings indicate that retinoic acid action is required during neuroectoderm induction for motoneuron specification and suggest that stem cells have restricted capacity to generate region-specific projection neurons even at an early developmental stage.