Long-acting PGE2 and Lisinopril Mitigate H-ARS.

Thrombocytopenia is a major complication in hematopoietic-acute radiation syndrome (H-ARS) that increases the risk of mortality from uncontrolled hemorrhage. There is a great demand for new therapies to improve survival and mitigate bleeding in H-ARS. Thrombopoiesis requires interactions between megakaryocytes (MKs) and endothelial cells. 16, 16-dimethyl prostaglandin E2 (dmPGE2), a longer-acting analogue of PGE2, promotes hematopoietic recovery after total-body irradiation (TBI), and various angiotensin-converting enzyme (ACE) inhibitors mitigate endothelial injury after radiation exposure. Here, we tested a combination therapy of dmPGE2 and lisinopril to mitigate thrombocytopenia in murine models of H-ARS following TBI. After 7.75 Gy TBI, dmPGE2 and lisinopril each increased survival relative to vehicle controls. Importantly, combined dmPGE2 and lisinopril therapy enhanced survival greater than either individual agent. Studies performed after 4 Gy TBI revealed reduced numbers of marrow MKs and circulating platelets. In addition, sublethal TBI induced abnormalities both in MK maturation and in in vitro and in vivo platelet function. dmPGE2, alone and in combination with lisinopril, improved recovery of marrow MKs and peripheral platelets. Finally, sublethal TBI transiently reduced the number of marrow Lin-CD45-CD31+Sca-1- sinusoidal endothelial cells, while combined dmPGE2 and lisinopril treatment, but not single-agent treatment, accelerated their recovery. Taken together, these data support the concept that combined dmPGE2 and lisinopril therapy improves thrombocytopenia and survival by promoting recovery of the MK lineage, as well as the MK niche, in the setting of H-ARS.

Impact of the calcium form of ß-hydroxy-ß-methylbutyrate upon human skeletal muscle protein metabolism.

BACKGROUND & AIMS: ß-hydroxy-ß-methylbutyrate (HMB) is purported as a key nutritional supplement for the preservation of muscle mass in health, disease and as an ergogenic aid in exercise. Of the two available forms of HMB (calcium (Ca-HMB) salt or free acid (FA-HMB)) - differences in plasma bioavailability have been reported. We previously reported that ∼3 g oral FA-HMB increased muscle protein synthesis (MPS) and reduced muscle protein breakdown (MPB). The objective of the present study was to quantify muscle protein metabolism responses to oral Ca-HMB. METHODS: Eight healthy young males received a primed constant infusion of 1,2 13C2 leucine and 2H5 phenylalanine to assess MPS (by tracer incorporation in myofibrils) and MPB (via arterio-venous (A-V) dilution) at baseline and following provision of ∼3 g of Ca-HMB; muscle anabolic (MPS) and catabolic (MPB) signalling was assessed via immunoblotting. RESULTS: Ca-HMB led a significant and rapid (<60 min) peak in plasma HMB concentrations (483.6 ± 14.2 µM, p < 0.0001). This rise in plasma HMB was accompanied by increases in MPS (PA: 0.046 ± 0.004%/h, CaHMB: 0.072 ± 0.004%/h, p < 0001) and suppressions in MPB (PA: 7.6 ± 1.2 µmol Phe per leg min-1, Ca-HMB: 5.2 ± 0.8 µmol Phe per leg min-1, p < 0.01). Increases in the phosphorylation of mTORc1 substrates i.e. p70S6K1 and RPS6 were also observed, with no changes detected in the MPB targets measured. CONCLUSIONS: These findings support the pro-anabolic properties of HMB via mTORc1, and show that despite proposed differences in bioavailability, Ca-HMB provides a comparable stimulation to MPS and suppression of MPB, to FA-HMB, further supporting its use as a pharmaconutrient in the modulation of muscle mass.

One-third of patients fail to return to work 1 year after surgery for colorectal cancer.

BACKGROUND: Achieving full recovery after colorectal cancer surgery means a return to normal physical and psychological health and to a normal social life. Recovery data focusses on time to discharge rather than longer term functionality including return to work (RTW). We aim to assess return to normal holistic function at 1 year after colorectal cancer surgery. METHOD: Questionnaires were created and dispatched to 204 patients who had undergone surgery with curative intent for colorectal cancer, in 2011-2012, in a single teaching hospital. RESULTS: Response rate was 75 % (153/204), 82 % (129/157) for open surgery (OS) and 51 % (24/47) for laparoscopic surgery (LS). Median age was 68 (48-91) years for OS and 65 (36-84) for LS. Eighty-four per cent of patients felt 'ready' and 95 % had adequate pain control upon discharge (no difference between groups). LS reported earlier 'return to full fitness' (1-3 months) than OS (>6 months; Mann-Whitney U, p < 0.05). Recovery from LS was 'better than expected' compared to OS 'worse than expected' (Mann-Whitney U test, p < 0.05). Forty-nine patients were employed preoperatively and 61 % (n = 30) returned to work. RTW was more frequent after LS (Chi-square test, p < 0.05). Length of time to RTW was significantly less after LS [44 (6-84) days] than OS [71 (14-252) days] (t test, p < 0.05). Levels of self-employment were equal between groups. CONCLUSIONS: One-third of patients failed to RTW at 1 year post-surgery. Patients having LS returned to full fitness faster, felt recovery was shorter and returned to work earlier than OS. We must invest more in managing expectations and provide better post-discharge support to improve RTW.

Effect of energy restriction and exercise on vitamin B-6 status of women during lactation.

PURPOSE: Lactation increases vitamin B-6 requirements because its concentration in breast milk is related to maternal intake and it is essential for infants. Exercise may also increase the requirement because it increases utilization and excretion of vitamin B-6. Therefore, the purpose of this study was to determine whether energy restriction and exercise affected vitamin B-6 status of lactating women. METHODS: Breastfeeding women with a body mass index > or = 25 and < or = 30 kg x m(-2) were randomly assigned at 4 wk postpartum to either restrict energy intake by 500 kcal x d(-1) and exercise for 45 min x d(-1), 4 d x wk(-1) (weight loss group, WG) or maintain usual diet and not exercise (control group, CG) for 10 wk. Women were given a supplement containing 2.0 mg of vitamin B-6. Measurements included vitamin B-6 concentrations in breast milk and plasma, plasma pyridoxal 5'-phosphate, and erythrocyte alanine aminotransferase activity. RESULTS: The WG lost more weight (-4.4 +/- 0.4 vs -0.9 +/- 0.5 kg, P < 0.01) than the CG. Cardiovascular fitness increased by 12% in the WG, compared to 3% in the CG (P = 0.09). Milk vitamin B-6 concentrations increased in both groups (161 +/- 107 and 191 +/- 85 nmol x L(-1), WG and CG, respectively, P = 0.05). There were no significant differences in other vitamin B-6 parameters. Weight and length gain (2.06 +/- 0.21 and 1.83 +/- 0.17 kg; 8.6 +/- 0.6 and 7.2 +/- 0.5 cm; WG and CG, respectively) of infants was not significantly different between groups. CONCLUSIONS: Energy restriction and exercise from 4 to 14 wk postpartum in overweight, breastfeeding women consuming adequate dietary intakes and 2.0 mg of supplemental vitamin B-6 does not adversely affect vitamin B-6 status or infant growth.

Cellular and molecular mechanisms of radiation inhibition of restenosis. Part I: role of the macrophage and platelet-derived growth factor.

PURPOSE: The major radiobiological issue in determining the rationale for the use of radiation to inhibit vascular restenosis is the identification of the target cell(s) and/or cytokine(s) responsible for neointimal hyperplasia and vascular remodeling. The central hypothesis of this report is that the macrophage/monocyte and PDGF are key elements in the process of neointimal hyperplasia seen following angioplasty, similar to their role in lesion formation and progression found in atherosclerotic thickening. Specific immunohistochemical and cytochemical stains were applied to a rat carotid model in a temporal series after balloon angioplasty to determine macrophage activity vs. smooth muscle cell proliferation, the latter being classically thought to be the cell responsible for restenosis. METHODS AND MATERIALS: Neointimal hyperplasia was created in an established rat carotid artery model by a balloon catheter technique. Immediately following injury, treatment groups received irradiation via high dose rate (HDR) brachytherapy, the 192Ir source being placed externally to the vessel. Radiation was delivered to a length of 2 cm of the injured vessel at doses of 5, 10, and 15 Gy, and the animals were sacrificed at various time points following treatment (24 h to 6 months). Serial sections of tissue were stained immunohistochemically with the primary antibodies CD11b, mac-1, anti-PDGF, and alpha-smooth muscle actin. RESULTS: Immediately (24 h) postinjury, there is an apparent migration of macrophages seen in the adventitia; after 1 week, proliferation and migration of macrophages could be seen clearly within all the vessel layers, especially in the intima; by 3 weeks, when there was evidence of neointimal hyperplasia, macrophages could still be seen, mainly in the intima scattered among the smooth muscle cells and myofibroblasts, and to a lesser degree at 6 months. There was corresponding expression of PDGF, whenever and wherever there were zones of activation/neointimal hyperplasia. Alpha-smooth muscle actin staining identified the smooth muscle cells distinct from the macrophages, and these SMCs exhibited activation in the neointimal hyperplasia zones at all later time points. Furthermore, we showed that radiation significantly reduced the macrophage population, while the onset of neointimal hyperplasia was accompanied by a return of the macrophage population. CONCLUSION: Our results suggest that the activated adventitial macrophage/monocyte are the key cells responsible for initiating the arterial neointimal hyperplasia and vascular remodeling developing postangioplasty as they are in the initiation and perpetuation of atheromatous thickening. Irradiation delivered immediately postinjury is, therefore, highly effective, because the macrophage population is exquisitely radiosensitive.

Blood glucose concentration does not affect outcome in brain trauma: A 31P MRS study.

Effects of blood glucose concentration on biochemical and neurologic outcome following lateral fluid percussion-induced traumatic injury of moderate severity (2.8 atm) in rats were studied using radioactive phosphorus (31P) magnetic resonance spectroscopy (MRS) and a battery of tests designed to evaluate posttraumatic neurologic motor function. Prior to injury, male Sprague-Dawley rats (n = 18) were randomly assigned to receive either dextrose, 2 ml 50% (wt/vol), zinc insulin (10 IU/kg) or no treatment, thus dividing the animals into hyperglycemic, hypoglycemic, and normoglycemic groups, respectively. Animals were then injured, monitored for 4 h by 31P MRS before being allowed to recover, and assessed for posttraumatic motor function. Following brain injury, there was no difference in brain intracellular pH between groups over the 4-h posttraumatic MRS monitoring period. Similarly, intracellular free magnesium, cytosolic phosphorylation potential, and neurologic outcome posttrauma were not significantly different between groups. We conclude that, unlike models of ischemia, blood glucose concentration may not be a significant factor affecting outcome in traumatic brain injury.

Bioenergetics and control of oxygen consumption in the in situ rat heart.

Control of respiration by products of ATP hydrolysis was examined in the in situ rat heart using a purpose-built nuclear magnetic resonance (NMR) coil. The in situ ratio of phosphocreatine to ATP concentrations ([PCr]/[ATP]) was 2.30 +/- 0.05, free Mg2+ concentration ([Mg2+]) was 0.57 mM, and cytosolic pH was 7.35 +/- 0.03 (n = 7). Basal inorganic phosphate concentration ([Pi]) was below NMR detection but was estimated to be 0.83 mM. The [ATP]/[ADP] [Pi] ratio, free ADP concentration ([ADP]), and free energy of ATP hydrolyses (delta GATP) were calculated to be 700,000 +/- 78,000 M-1, 18 +/- 3 microM, and -63.93 +/- 0.33 kJ/mol in situ, respectively (n = 7). In contrast, in the Langendorff perfused rat heart [ATP]/[ADP] [Pi] was only 76,140 +/- 12,830 M-1, [ADP] was 65 +/- 9 microM, and delta GATP was -59.92 +/- 0.48 kJ/mol (n = 7), all indicative of a lower energy state in vitro. Epinephrine infusion in situ (0.9 microgram.min-1.kg-1) increased the rate-pressure product 2.05-fold. During stimulation [ATP] was stable at 97 +/- 3% signal intensity, [PCr] declined by 25%, and [Pi] increased to 1.83 mM. Cytosolic pH was 7.27 +/- 0.01 and [Mg2+] was 0.64 +/- 0.05 mM. [PCr]/[ATP] declined to 1.83 +/- 0.13, and [ATP]/[ADP] [Pi] fell to 108,000 +/- 15,000 M-1. delta GATP only fell marginally to -59.56 +/- 0.49 kJ/mol. Free [ADP] increased threefold to 55 +/- 10 microM. Infusion of 2.8 +/- 0.5 microgram.min-1.kg-1 epinephrine increased the rate-pressure product 2.7-fold, further reduced [ATP]/[ADP] [Pi] (5% of basal), and elevated [ADP] more than fourfold without changing [ATP]. We conclude that the in situ heart is highly energetic compared with isolated perfused hearts and operates at a different metabolic "set-point." Because free [ADP] and [Pi] in situ approximate apparent Michaelis constants for mitochondrial respiration in vitro and increase with increased cardiac work, we conclude that each fulfills the criteria for the kinetic control of O2 consumption in the in situ rat myocardium.

Changes in the 14C-labelling of molecular species of 3-monogalactosyl-1,2-diacylglycerol in leaves of Vicia faba treated with compound San 9785.

The effects of the substituted pyridazinone herbicide. San 9785, on the biosynthesis of monogalactosyldiacylglycerol (MGDG) molecular species and on the diacylglycerol precursors were studied. Kinetic experiments with [14C]glucose-infiltrated Vicia faba leaf tissue showed that San 9785 inhibited desaturation of MGDG linoleic acid (C18:2) to form linolenic acid (C18:3) and increased the degree of unsaturation of the diacylglycerol molecular species used in MGDG biosynthesis. These results confirmed that the diacylglycerol precursor of MGDG contains highly unsaturated fatty acids, particularly C18:2 and C18:3. The results also indicated that the MGDG 3/3 molecular species (a molecular species of MGDG containing two C18:3 molecules) is derived from MGDG 2/3 (i.e. containing one C18:2 and one C18:3 fatty acid moiety) and, likely, MGDG 2/2 (i.e. containing two C18:2 fatty acid moieties), via sequential fatty acid desaturation in situ. A model for MGDG 3/3 biosynthesis in 'C18:3-plants', incorporating several sites for fatty acid desaturation (in the phospholipid and diacylglycerol precursors, which are not inhibited by San 9785, and in MGDG, which is inhibited by San 9785) is discussed.

Spontaneous thrombosis of deep cerebral veins: a complication of arteriovenous malformation.

An uncommon type of stroke in children is presented. An intracranial arteriovenous malformation in a 13-year-old boy spontaneously occluded about 22 months after surgical intervention. Precipitating factors, such as bacterial infections, could not be demonstrated in this patient, who had been attending school since the time of the craniotomy. The histological features of venous encephalomalacia in the galenic territory are contrasted with hemorrhagic encephalomalacia as seen after arterial occlusions: in the former, hemorrhages are more widespread and edema is more pronounced.