Effects of dietary vitamin E concentration and source on sow, milk, and pig concentrations of α-tocopherol.

A total of 126 gilts and sows (PIC 1050) and their litters were used to determine the effects of dietary vitamin E concentration and source on sow plasma, milk, and pig concentrations of α-tocopherol. Additionally, we estimated the bioavailability of D-α-tocopheryl acetate (D-α-TAc) relative to DL-α-tocopheryl acetate (DL-α-TAc) when fed in diets containing dried distillers grains with solubles (DDGS). The 6 dietary treatments included DL-α-TAc at 44 and 66 mg/kg and D-α-TAc at 11, 22, 33, and 44 mg/kg. From breeding to d 69 of gestation, sows were fed 2.0 kg/d of a diet containing 40% DDGS, 0.30 mg/kg added Se, and no added vitamin E. Vitamin E treatments were fed from d 70 of gestation through weaning. Plasma was collected from sows on d 69 and 100 of gestation, at farrowing, and at weaning. Colostrum and milk samples were also collected. Plasma from 3 pigs per litter and heart and liver samples from 1 pig per litter were collected at weaning. Plasma, milk, and tissues from 6 litters per treatment were analyzed for α-tocopherol. Although tissue, plasma, and milk concentrations of α-tocopherol were the primary response criteria of interest, sow and litter performance were measured. As expected, treatment effects were not observed for lactation feed intake, sow BW, or backfat measurements. A trend (P = 0.085) for a treatment effect on average pig BW at weaning was detected, with pigs nursing sows fed 44 mg/kg DL-α-TAc weighing less because of a younger weaning age. No other differences in litter performance were observed. As D-α-TAc increased in the diet, sow plasma, colostrum, and milk, pig plasma, and pig heart concentrations of α-tocopherol increased (linear, P < 0.03). Sows fed diets with 44 mg/kg D-α-TAc had increased (P < 0.03) plasma and colostrum and pig plasma concentrations of α-tocopherol compared with sows fed 44 mg/kg of DL-α-TAc. Sows fed 66 mg/kg DL-α-TAc also had greater (P = 0.022) plasma α-tocopherol at weaning than sows fed 44 mg/kg DL-α-TAc. Bioavailability coefficients for D-α-TAc relative to DL-α-TAc ranged from 1.9 to 4.2 for sow and pig plasma α-tocopherol, 2.9 to 3.6 for colostrum α-tocopherol, 1.6 for milk α-tocopherol, and 1.7 to 2.0 for pig heart and liver α-tocopherol. Overall, this study indicates the bioavailability for D-α-TAc relative to DL-α-TAc varies depending on the response criteria but is greater than the standard potency value of 1.36.

Exome sequencing of pleuropulmonary blastoma reveals frequent biallelic loss of TP53 and two hits in DICER1 resulting in retention of 5p-derived miRNA hairpin loop sequences.

Pleuropulmonary blastoma is a rare childhood malignancy of lung mesenchymal cells that can remain dormant as epithelial cysts or progress to high-grade sarcoma. Predisposing germline loss-of-function DICER1 variants have been described. We sought to uncover additional contributors through whole exome sequencing of 15 tumor/normal pairs, followed by targeted resequencing, miRNA analysis and immunohistochemical analysis of additional tumors. In addition to frequent biallelic loss  of TP53 and mutations of NRAS or BRAF in some cases, each case had compound disruption of DICER1: a germline (12 cases) or somatic (3 cases) loss-of-function variant plus a somatic missense mutation in the RNase IIIb domain. 5p-Derived microRNA (miRNA) transcripts retained abnormal precursor miRNA loop sequences normally removed by DICER1. This work both defines a genetic interaction landscape with DICER1 mutation and provides evidence for alteration in miRNA transcripts as a consequence of DICER1 disruption in cancer.

Omalizumab therapy is associated with reduced circulating basophil populations in asthmatic children.

Basophils have been implicated in promoting the early development of TH 2 cell responses in some murine models of TH 2 cytokine-associated inflammation. However, the specific role of basophils in allergic asthma remains an active area of research. Recent studies in animal models and human subjects suggest that IgE may regulate the homeostasis of human basophil populations. Here, we examine basophil populations in children with severe asthma before and during therapy with the IgE-directed monoclonal antibody omalizumab. Omalizumab therapy was associated with a significant reduction in circulating basophil numbers, a finding that was concurrent with improved clinical outcomes. The observation that circulating basophils are reduced following omalizumab therapy supports a mechanistic link between IgE levels and circulating basophil populations, and may provide new insights into one mechanism by which omalizumab improves asthma symptoms.

Minimum volume of local anaesthetic required to surround each of the constituent nerves of the axillary brachial plexus, using ultrasound guidance: a pilot study.

BACKGROUND: The minimum effective volume of local anaesthetic needed to provide effective analgesia of the four main branches of the axillary brachial plexus is unknown. This study was performed to determine the minimum volume of local anaesthetic required to surround the nerves of the axillary brachial plexus and document onset and duration of sensory and motor effects. METHODS: We enrolled 19 ASA I-II patients undergoing hand or forearm surgery. The four nerves of the axillary plexus were identified with ultrasound guidance. Lidocaine 1.5% with epinephrine 1:200 000 was loaded into a syringe driver. A 22 G needle was inserted in the long axis to each nerve and injection commenced using the bolus function (600 ml h(-1)). The needle was repositioned until the nerve was completely surrounded. The bolus dose in millilitres displayed on the syringe driver was recorded. This was repeated for each nerve. The degree of sensory and motor block was recorded as secondary outcomes. RESULTS: The mean (95% CI) volume to surround each nerve was: radial 3.42 (2.84-3.99) ml, median 2.75 (2.31-3.19) ml, ulnar 2.58 (2.14-3.03) ml, and musculocutaneous 2.30 (1.96-2.64) ml. The mean (95% CI) onset time for complete sensory block was: radial 22.5 (13.5-31.5) min, median 26.8 (18.5-35.0) min, ulnar 26.6 (17.8-35.4) min, and musculocutaneous 15.8 (7.45-24.2) min. The mean (95% CI) last recorded time with complete block was: radial 137.1 (105.6-168.7) min, median 144.7 (123.4-166.0) min, ulnar 183.2 (158.1-208.2) min, and musculocutaneous 158.3 (131.8-184.9) min. Seven patients required additional local anaesthetic infiltration and two required i.v. analgesia. No patient required conversion to general anaesthesia for surgery. CONCLUSIONS: We found that it is possible to surround each nerve of the axillary brachial plexus with 2-4 ml of local anaesthetic. We speculate that increasing this volume would produce blocks of quicker onset and longer duration while still using smaller volumes than previously thought.

Metagenomic analyses reveal antibiotic-induced temporal and spatial changes in intestinal microbiota with associated alterations in immune cell homeostasis.

Despite widespread use of antibiotics, few studies have measured their effects on the burden or diversity of bacteria in the mammalian intestine. We developed an oral antibiotic treatment protocol and characterized its effects on murine intestinal bacterial communities and immune cell homeostasis. Antibiotic administration resulted in a 10-fold reduction in the amount of intestinal bacteria present and sequencing of 16S rDNA segments revealed significant temporal and spatial effects on luminal and mucosal-associated communities including reductions in luminal Firmicutes and mucosal-associated Lactobacillus species, and persistence of bacteria belonging to the Bacteroidetes and Proteobacteria phyla. Concurrently, antibiotic administration resulted in reduced RELM beta production, and reduced production of interferon-gamma and interleukin-17A by mucosal CD4(+) T lymphocytes. This comprehensive temporal and spatial metagenomic analyses will provide a resource and framework to test the influence of bacterial communities in murine models of human disease.

Effect of vitamin E source, natural versus synthetic, and quantity on serum and tissue alpha-tocopherol concentrations in finishing swine.

Relative vitamin E status of pigs fed natural or synthetic vitamin E was evaluated based on serum and tissue alpha-tocopherol concentrations. Individually fed finishing gilts at a BW of 70.5 kg (n = 24) were allotted to dietary treatments based on initial BW. The 5 dietary treatments consisted of a positive control diet using synthetic vitamin E acetate (Syn E Ac) supplemented at 22 mg/kg, and 4 dietary levels of natural vitamin E acetate (Nat E Ac) supplemented at 6.71, 8.33, 11.00, and 16.18 mg/kg of diet. Before initiation of the 32-d experiment, pigs were fed a non-vitamin E-fortified diet for 30 d. Diets were formulated to contain true ileal digestible lysine of 0.9 and 0.8% for the pretest and test diets. Serum samples were collected on d 15 and 32, whereas tissue samples were collected on d 32 for alpha-tocopherol analysis. Serum alpha-tocopherol concentrations on d 15 and 32 were greater (P < 0.05) in pigs fed 8.33, 11.00, or 16.18 mg/kg of Nat E Ac than in pigs fed 22 mg/kg of Syn E Ac. When compared with pigs fed 22 mg/kg of Syn E Ac, alpha-tocopherol concentrations were greater (P < 0.05) in 6 tissues (heart, kidney, spleen, liver, lung, and adipose) in pigs fed 16.18 mg/kg of Nat E Ac; greater (P < 0.05) in heart, kidney, spleen, liver, and adipose tissue in pigs fed 11.00 mg/kg of Nat E Ac; and greater (P < 0.05) in spleen, loin, and adipose tissue in pigs fed 8.33 mg/kg of Nat E Ac. As dietary Nat E Ac increased from 6.71 to 16.18 mg/kg, serum alpha-tocopherol increased linearly (P < 0.01) on d 15 and 32 of the experiment. Increasing dietary Nat E Ac linearly increased (P < 0.05) alpha-tocopherol concentrations for lung, heart, kidney, spleen, and liver. These results indicate that Nat E Ac was an effective vitamin E source and its relative bioavailability was substantially greater than 1.36 for finishing swine when compared with Syn E Ac.

An evaluation of natural (RRR-alpha-tocopheryl acetate) and synthetic (all-rac-alpha-tocopheryl acetate) vitamin E fortification in the diet or drinking water of weanling pigs.

Three experiments conducted with weanling pigs evaluated the effects of vitamin E added to the drinking water or diet on plasma and tissue alpha-tocopherol concentrations. When natural or synthetic vitamin E was used, it was added at an IU-equivalent basis, but natural vitamin E was 73.5% (mg basis) of the synthetic vitamin E. Experiment 1 used 18-d-old weanling pigs (n = 120) in a 3 x 2 factorial arrangement of treatments in a randomized complete block design with 4 replicates. The first factor evaluated the dietary levels of natural vitamin E (RRR-alpha-tocopheryl acetate) added at 0, 50, or 300 IU/kg, whereas the second factor was the natural vitamin E added to the drinking water at 0 or 100 IU/L. Pigs were bled at periodic intervals, and 1 pig per pen was killed at the end of the 21-d trial and tissues (liver, heart, lung, and loin) were collected for alpha-tocopherol analysis. When vitamin E was not added to the diet or water, plasma alpha-tocopherol declined over the 21-d period. Although there were some interactions (P < 0.01), tissue and plasma alpha-tocopherol concentrations increased linearly when vitamin E was added to the diet or water. Experiment 2 was a 3 x 2 factorial in a randomized complete block design with 4 replicates. A total of 96 pigs weaned at 18 d of age, with an initial BW of 6.2 kg, were fed a nonvitamin E fortified diet, but natural or synthetic (all-rac-alpha-tocopheryl acetate) vitamin E was added to their drinking water at 50, 100, or 150 IU/L. Pigs were bled at 0, 3, 7, 10, 14, and 21 d postweaning, with tissues (liver, lung, heart, and loin) collected for alpha-tocopherol analysis at d 21. The results indicated that plasma alpha-tocopherol concentrations increased (P < 0.01) as vitamin E increased, with greater tissue alpha-tocopherol concentrations (P < 0.01) when natural vitamin E was provided. Experiment 3 was conducted in 2 replicates, but pigs (n = 60) were not provided vitamin E in the diet or water for 7 d postweaning, and then natural or synthetic vitamin E was added to the drinking water as in Exp. 2 (50, 100, or 150 IU/L). Pigs were bled at 0, 2, 4, 6, 8, 10, and 24 h after being provided vitamin E to evaluate the absorption from each vitamin E source and level. Plasma alpha-tocopherol increased quadratically (P < 0.01) and plateaued at 8 to 10 h for each treatment group. These results indicate that adding vitamin E to the pig's water supply at weaning was more effective in increasing plasma alpha-tocopherol than when it was added to the diet during the initial 14 d postweaning, and that natural vitamin E was a superior source compared with synthetic vitamin E.

Sciatica: a review of history, epidemiology, pathogenesis, and the role of epidural steroid injection in management.

Radicular pain in the distribution of the sciatic nerve, resulting from herniation of one or more lumbar intervertebral discs, is a frequent and often debilitating event. The lifetime incidence of this condition is estimated to be between 13% and 40%. Fortunately, the majority of cases resolve spontaneously with simple analgesia and physiotherapy. However, the condition has the potential to become chronic and intractable, with major socio-economic implications. This review discusses the history, epidemiology, pathophysiology, and natural history of sciatica. A Medline search was performed to obtain the published literature on the sciatica, between 1966 and 2006. Hand searches of relevant journals were also performed. Epidemiological factors found to influence incidence of sciatica included increasing height, age, genetic predisposition, walking, jogging (if a previous history of sciatica), and particular physical occupations, including driving. The influence of herniated nucleus pulposus and the probable cytokine-mediated inflammatory response in lumbar and sacral nerve roots is discussed. An abnormal immune response and possible mechanical factors are also proposed as factors that may mediate pain. The ongoing issue of the role of epidural steroid injection in the treatment of this condition is also discussed, as well as potential hazards of this procedure and the direction that future research should take.

Internalization of near-infrared fluorescent dyes within isolated macrophage populations.

The development and application of microsensor technology has enhanced the ability of scientists to further understand various biological activities, such as changes in the intracellular environment after injury or toxic exposure. NIR microsensor technology may be useful in detecting the cellular injuries or adverse changes during the early onset period, allowing for the administration of therapies to initiate recovery. The development and use of Infrared (IR) and near infrared (NIR) dyes as biological micro-sensors due to their advanced spectral characteristics may be helpful. Three of the more useful NIR dye characteristics include the ability to minimize background interference by extraneous biological matrices, the ability to exhibit optimal molar absorptivity and quantum yields, and the ability to maintain normal cellular activity. Thus, the current study was designed to investigate the ability of selected NIR micro-sensor dyes to undergo cellular internalization, demonstrate intracellular NIR fluorescent signaling, and maintain normal cellular activity. The results demonstrate that the selected NIR micro-sensor dyes undergo cellular internalization. The presence of the dyes within the cells did not affect cell viability. In addition, these dyes demonstrate changes in absorbance and fluorescence after the immune cells were challenged with a stimulant. Moreover, critical cellular functions, such as tumor necrosis factor release and superoxide production were not compromised by the internalization of the fluorescent dyes. These data suggest that selected NIR micro-sensor dyes can undergo intracellular internalization within isolated macrophages without adversely affecting various parameters of normal cellular activity.

Patient controlled analgesia for labour: a comparison of remifentanil with pethidine.

We compared the analgesic efficacy and safety of remifentanil and pethidine via patient controlled analgesia for women in established uncomplicated labour. Women received either remifentanil 40 microg with a 2-min lockout (n = 20) or pethidine 15 mg with a 10-min lockout (n = 19). Visual analogue scores for pain during the study and for overall pain were similar for both groups (mean (SD) 6.4 (1.5) cm for remifentanil and 6.9 (1.7) cm for pethidine). The area under the curve for visual analogue scores of satisfaction with analgesia was higher for remifentanil than for pethidine (p = 0.001). Maternal arterial oxygen saturation was similar in both groups. Neurologic and Adaptive Capacity Scores at 30 min were higher for remifentanil than for pethidine (median (interquartile range [range]) 36 (34.5-37 [32-39]) vs 34 (33-35 [30-35]), respectively; p = 0.003).

Assessment of tracheal intubation in children after induction with propofol and different doses of remifentanil.

Tracheal intubating conditions were assessed in 112 children after induction of anaesthesia with propofol and remifentanil 1.0, 2.0 or 3.0 micro g.kg-1. Subjects in a control group were given propofol and mivacurium 0.2 mg.kg-1. Haemodynamic and respiratory parameters were recorded. Plasma catecholamine levels were measured in a subgroup of 40 children. Intubating conditions were acceptable in 14/28 (50%), 18/26 (69%) and 22/27 (82%) in those subjects given remifentanil 1.0, 2.0 or 3.0 micro g.kg-1, respectively, and in 27/28 (96%) of the control group. Intubating conditions in subjects given remifentanil 3.0 micro g.kg-1 were better than in those given remifentanil 1.0 micro g.kg-1 (p < 0.05). There were no significant differences in intubating conditions between those given remifentanil 3.0 micro g.kg-1 and the control group. Systolic blood pressure and heart rate increased in response to tracheal intubation in subjects given remifentanil 1.0 micro g.kg-1 and in the control group (p < 0.05). Time to resumption of spontaneous respiration was prolonged in subjects given remifentanil 3.0 micro g.kg-1 (p < 0.001). In conclusion, remifentanil 2 micro g.kg-1 provides acceptable intubating conditions and haemodynamic stability without prolonging the return of spontaneous respiration.

Neonatal alveolar rhabdomyosarcoma with skin and brain metastases.

BACKGROUND: Approximately 5-10% of patients with rhabdomyosarcomas (RMS) are diagnosed during the first year of life, and their clinical characteristics have been well documented. However, because RMS rarely occurs during the neonatal period, little is known about neonatal RMS. METHODS: Four patients with neonatal RMS were treated at St. Jude Children's Research Hospital between 1962 and 1999. The authors report the results of a review of these patients and of cases described in the literature. Clinical, radiologic, and pathologic features of these patients and their outcomes were evaluated. RESULTS: One patient with embryonal RMS was treated successfully with a combination of systemic chemotherapy and local control measures. The other three patients had alveolar RMS. Two of them had multiple skin and subcutaneous metastatic nodules at the time of diagnosis and developed brain metastases early in their course. In one of these patients, the PAX3-FKHR fusion transcript was detected. Three other similar cases of neonatal alveolar RMS with metastases to the skin and brain have been reported in the literature. CONCLUSIONS: A distinct syndrome of neonatal RMS is described. This syndrome is characterized by alveolar histology, multiple skin and subcutaneous metastases, and fatal outcome as the result of early brain metastasis.

Anaplastic lymphoma kinase (ALK) expression in the inflammatory myofibroblastic tumor: a comparative immunohistochemical study.

Inflammatory myofibroblastic tumor (IMT) is an uncommon mesenchymal neoplasm with a variable histologic appearance that may mimic other spindle cell processes, particularly nodular fasciitis, desmoid tumor, and in intra-abdominal locations, gastrointestinal stromal tumor. Recently, gene fusions involving ALK at chromosome 2p23 have been described in IMTs. The resultant ALK protein overexpression in the myofibroblastic component of these tumors is detectable by immunohistochemistry. We examined 73 IMTs, 20 cases of nodular fasciitis, 15 desmoid fibromatoses, and 15 gastrointestinal stromal tumors by immunohistochemistry using ALK-11, a rabbit polyclonal antibody that recognizes the C-terminus of the protein. ALK positivity was detected in 44 of 73 (60%) IMTs. All cases of nodular fasciitis, desmoid fibromatosis, and gastrointestinal stromal tumors were ALK negative (p < 0.001). These findings demonstrate that ALK positivity is common in IMTs, and immunohistochemistry using anti-ALK antibodies can be helpful in the differential diagnosis of these neoplasms. In addition, anti-ALK staining seems to correlate with those IMTs that have the typical tri-patterned histologic appearance and clinical presentation, providing additional support to the premise that IMT is a distinctive clinicopathologic entity within the broad category of inflammatory pseudotumors.

Molecular genetic aspects of solid tumors in childhood.

Advances in molecular genetic research in the past 2 decades have led to an increased understanding of the genetic events in the pathogenesis and progression of human malignancies, including those of childhood. A number of pediatric malignancies have served as models for the molecular genetic approach to patients with cancer. These have highlighted the utility of molecular analysis for a variety of purposes including diagnosis, risk stratification and treatment planning, understanding of syndromes associated with cancer and genetic screening, genetic counseling and prophylactic treatment including surgery. It is likely that there soon will be individualized treatment regimens based on the molecular biologic profile of a patient's tumor. In addition, molecular profiling will lead to new drug development designed to induce differentiation of tumor cells, block dysregulated growth pathways, or reactivate silenced apoptotic pathways. This review discusses the molecular genetic aspects of some of the more common pediatric tumors as well as tumors whose pathogenetic mechanisms are particularly instructive.

Fusion of the ALK gene to the clathrin heavy chain gene, CLTC, in inflammatory myofibroblastic tumor.

Inflammatory myofibroblastic tumor (IMT) is a rare, but distinctive mesenchymal neoplasm composed of fascicles of bland myofibroblasts admixed with a prominent inflammatory component. Genetic studies of IMTs have demonstrated chromosomal abnormalities of 2p23 and rearrangement of the anaplastic lymphoma kinase (ALK) gene locus. In a subset of IMTs, the ALK C-terminal kinase domain is fused with a tropomyosin N-terminal coiled-coil domain. In the current study, fusion of ALK with the clathrin heavy chain (CTLC) gene localized to 17q23 was detected in two cases of IMT. One of these cases exhibited a 2;17 translocation in addition to other karyotypic anomalies [46,XX,t(2;17)(p23;q23),add(16)(q24)].

Patient-controlled analgesia for labour using remifentanil: a feasibility study.

We have investigated the efficacy and safety of remifentanil in a patient-controlled analgesia device for labour in 21 women. Remifentanil was available in increasing doses (bolus doses 0.25-1.0 microg x kg(-1)) with and without a background infusion (0.025-0.05 microg x kg(-1) x min(-1)). A lockout time of 2 min was used. Thirteen out of 21 (62%) women chose to continue using remifentanil up to and during delivery. Nineteen out of 21 (90%) achieved a reduction in pain score from baseline. Using a VAS of 0-10 cm the median maximum reduction in pain score was 3 cm (range 0-8 cm). There was a significant reduction (P<0.05) from baseline pain scores (median= 8 cm) to scores at bolus doses in the range 0.25-0.5 microg x kg(-1) (median=5 cm). There were no significant reductions in the fetal heart rate. Apgar scores and cord blood gas analyses remained within normal limits. We conclude that a remifentanil patient-controlled analgesia system (bolus doses 0.25-0.5 microg x kg(-1), without a background infusion) may safely provide worthwhile, although incomplete, analgesia for labour.

Influence of linker histone H1 on chromatin remodeling.

Chromatin-remodeling complexes have been a central area of focus for research dealing with accessing cellular DNA sequestered in chromatin. Although the linker histone H1 plays a major role in promoting and maintaining higher-order chromatin structure, it has been noticeably absent from assays utilizing chromatin-remodeling enzymes. This review focuses on two ATP-dependent chromatin-remodeling complexes, Drosophila ISWI and mammalian SWI/SNF, that have been assayed using chromatin templates containing histone H1.

Secondary ovarian neoplasms in children: imaging features with histopathologic correlation.

BACKGROUND: Although the pathologic features and imaging appearance of childhood primary ovarian neoplasms have been well described, little information is available about the malignancies that may secondarily involve the ovary. OBJECTIVE: To determine the relationship between the imaging features and the histopathology of secondary ovarian neoplasms in children treated at our institution. MATERIALS AND METHODS: We searched our institutional database for codes indicating metastatic ovarian disease. Of the 35 patients with such codes, 18 had pathologically proven secondary ovarian disease. From their medical records we recorded demographic data, presenting symptoms, and evidence of endocrine dysfunction. We reviewed the pre-oophorectomy imaging and the subsequent pathologic specimens. RESULTS: One-third of the patients had bilateral pelvic masses; another third had large masses indistinguishable from the ovaries. Twelve (67%) had either ascites, peritoneal implants, matted bowel, adenopathy, pleural effusions, or some combination of these. Five (28%) had other metastatic disease. Primary tumors included colon adenocarcinoma (9), Burkitt's lymphoma (3), alveolar rhabdomyosarcoma (3), Wilms' tumor (1), neuroblastoma (1), and retinoblastoma (1). CONCLUSION: Although rare, secondary ovarian tumors should be considered in the differential diagnosis of children with ovarian masses. Bilateral ovarian masses or large masses indistinguishable from the ovaries, particularly in the presence of other metastatic foci, may help distinguish primary from secondary ovarian malignancies.