Quantitative trait loci mapping of polyphenol metabolites from a 'Latham' x 'Glen Moy' red raspberry (Rubus idaeus L) cross.

OBJECTIVE: The objective of this study was to investigate the genetic control of polyphenol accumulation in red raspberry (Rubus idaeus L). METHODS: The levels of total anthocyanins and 37 individual polyphenol metabolites were measured over three years in a raspberry biparental mapping population. Quantitative trait loci (QTLs) for these traits were mapped onto a high-density SNP linkage map. RESULTS: At least one QTL was detected for each trait, with good consistency among the years. On four linkage groups (LG), there were major QTLs affecting several metabolites. On LG1, a QTL had large effects on anthocyanins and flavonols containing a rutinoside or rhamnose group. On LG4, a QTL had large effects on several flavonols and on LG5 and LG6 QTLs had large effects on ellagic acid derivatives. Smaller QTLs were found on LG2 and LG3. CONCLUSION: The identification of robust QTLs for key polyphenols in raspberry provides great potential for marker-assisted breeding for improved levels of potentially health beneficial components.

Persistence of HIV reservoir following successful haematopoietic stem cell transplant for juvenile myelomonocytic leukaemia in a child with perinatally acquired HIV.

This report describes a case of juvenile myelomonocytic leukaemia (JMML) on a background of both perinatally acquired HIV infection and congenital cytomegalovirus, and management of antiretroviral therapy during haematopoietic stem cell transplant. Peripheral blood HIV viral load remained below the lower limit of detection throughout and following transplant and is currently <20 RNA copies/mL. The child is currently in remission from JMML, but HIV DNA remains detectable despite myeloablative conditioning and sustained plasma HIV viral suppression.

De novo intracerebral arteriovenous malformations and a review of the theories of their formation.

PURPOSE: Arteriovenous malformations (AVM) are still frequently described as congenital lesions in medical texts despite little evidence existing for their congenital nature. Increasing numbers of case reports of de novo AVMs add weight to the notion that they are dynamic lesions and that they can form postnatally. A thorough review of all reported cases of de novo AVM formation and a review of articles relating to AVM pathogenesis was planned to summarise current research on AVM pathogenesis and provide insight into the future implications for AVM research and treatment. METHODS AND RESULTS: MEDLINE was searched to find 29 cases of de novo AVM formation with prior MRI imaging, nine of which also had prior digital subtraction angiography. A discussion of AVM pathogenesis is undertaken through a review of articles relating to AVM embryology, postnatal angiogenesis, syndromic forms of AVMs and studies of AVM molecular biology and genetics in human and animal models. CONCLUSIONS: There is little evidence for an embryological origin through dysregulated vasculogenesis, whereas there is a raft of evidence to support dysregulated angiogenesis in childhood or even adulthood. Translational implications include risk stratification by biomarkers for predicting haemorrhage and novel therapeutic approaches to suppress AVM proliferation and initiate reversal.

Defining the osteoarthritis patient: back to the future.

The history of osteoarthritis (OA) is important because it can help broaden our perspective on past and present controversies. The naming of OA, beginning with Heberden's nodes, is itself a fascinating story. According to Albert Hoffa, R. Llewellyn Jones and Archibald Edward Garrod, the name OA was introduced in the mid-nineteenth century by surgeon Richard von Volkmann who distinguished it from rheumatoid arthritis and gout. Others preferred the terms 'chronical rheumatism', 'senile arthritis', 'hypertrophic arthritis' or 'arthritis deformans'. A similar narrative applies to the concept of OA affecting the whole joint vs the 'wear-and-tear' hypothesis, inflammation and the role of the central nervous system (CNS). In the late nineteenth and early twentieth centuries, the Garrods (father and son) and Hermann Senator argued that OA was a whole joint disease, and that inflammation played a major role in its progression. Garrod Jnr and John Spender also linked OA to a neurogenic lesion 'outside the joint'. The remaining twentieth century was no less dynamic, with major advances in basic science, diagnostics, treatments, surgical interventions and technologies. Today, OA is characterized as a multi-disease with inflammation, immune and CNS dysfunction playing central roles in whole joint damage, injury progression, pain and disability. In the current 'omics' era (genomics, proteomics and metabolomics), we owe a great debt to past physicians and surgeons who dared to think 'outside-the-box' to explain and treat OA. Over 130 years later, despite these developments, we still don't fully understand the underlying complexities of OA, and we still don't have a cure.

Epidural abscess with associated spondylodiscitis following prostatic biopsy.

Spondylodiscitis is often iatrogenic in nature. We report the case of a 69-year-old man presenting with spondylodiscitis and associated epidural abscess following transrectal ultrasonography guided prostate biopsy despite ciprofloxacin cover. To our knowledge, this is the first case of spondylodiscitis secondary to fluoroquinolone resistant Escherichia coli.

Pulmonary function after hemorrhagic shock and resuscitation in a porcine model.

BACKGROUND: Hemorrhagic shock may trigger an inflammatory response and acute lung injury. The combination adenosine, lidocaine (AL) plus Mg(2+) (ALM) has organ-protective and anti-inflammatory properties with potential benefits in resuscitation.The aims of this study were to investigate: (1) pulmonary function and inflammation after hemorrhagic shock; (2) the effects of ALM/AL on pulmonary function and inflammation. METHODS: Pigs (38 kg) were randomized to: sham + saline (n = 5); sham + ALM/AL (n = 5); hemorrhage control (n = 11); and hemorrhage + ALM/AL (n = 9). Hemorrhage animals bled to a mean arterial pressure (MAP) of 35 mmHg for 90 min, received resuscitation with Ringer's acetate and 20 ml of 7.5% NaCl with ALM to a minimum MAP of 50 mmHg, after 30 min shed blood and 0.9% NaCl with AL were infused. Hemorrhage controls did not receive ALM/AL. Primary endpoints were pulmonary wet/dry ratio, PaO2 /FiO2 ratio (partial pressure of arterial oxygen to the fraction of inspired oxygen), cytokine and protein measurements in bronchoalveolar lavage fluid (BALF) and lung tissue, neutrophil invasion and blood flow in lung tissue. RESULTS: In the hemorrhage groups, wet/dry ratio increased significantly compared with the sham groups. PaO2 /FiO2 ratio decreased during shock but normalized after resuscitation. BALF did not indicate significant pulmonary inflammation, oxidative stress or increased permeability. Intervention with ALM caused a temporary increase in pulmonary vascular resistance and reduced urea diffusion across the alveolar epithelia, but had no effect on wet/dry ratio. CONCLUSION: Hemorrhagic shock and resuscitation did not cause acute lung injury or pulmonary inflammation. The question whether ALM/AL has the potential to attenuate acute lung injury is unanswered.

Physiological and metabolic effects of prophylactic treatment with the osmolytes glycerol and betaine on Bos indicus steers during long duration transportation.

The physiological and metabolic effects of prophylactic treatment with osmolytes were investigated using twenty-four 2.5-yr-old Bos indicus steers. Animals were allocated to 1 of 4 treatment groups: 1) control, feed and water deprived for 48 h (n = 6); 2) transported, transported for 48 h (n = 6); 3) glycerol, dosed with glycerol (2 g/kg of BW) and then transported for 48 h (n = 6); and 4) betaine, dosed with betaine (0.25 g/kg of BW) then transported for 48 h (n = 6). Body water, electrolytes, blood pH and gases, plasma lactate, glucose, albumin, total protein, anion gap, strong ion difference, total weak acids, and BW were determined at the conclusion of 24 and 48 h of transportation. The glycerol group had greater body water volumes than the control (P = 0.05) and transported (P = 0.02) groups. The glycerol, transported, and betaine groups had lower (P = 0.02) plasma Mg concentrations than the control group at 24 h, whereas the glycerol group maintained lower (P = 0.04) plasma concentrations of Ca than the control group. The betaine group had lower (P = 0.04) hematocrit than the control group at 24 and 48 h. Plasma bicarbonate and pCO2 were 13 and 17% greater (P = 0.01 and 0.04, respectively) in the glycerol group at 24 h compared with control and transported groups. However, the ratio of [HCO3]/[CO2] in the glycerol group did not differ from the other groups and thereby maintained pH. The glycerol group maintained a 30% greater (P < 0.001) plasma concentration of glucose than the control group, and 14% greater (P = 0.05) than the transported and betaine groups. In contrast, betaine had little effect on increasing blood glucose compared with glycerol. Glycerol-linked hyperhydration at 24 h may not only help to conserve water loss during long distance transportation, but the increased blood glucose may have an important protein-sparing effect due, in part, to greater insulin concentrations inhibiting the breakdown of muscle proteins, thus, countering the amino-acid mobilizing effect of cortisol after 24 h. Therefore, the osmolyte glycerol shows promise as a prophylactic treatment for attenuating the effects of long distance transportation by maintaining body water, decreasing the energy deficit, and preserving health and muscle quality.

Endografting of the descending thoracic aorta increases ascending aortic input impedance and attenuates pressure transmission in dogs.

OBJECTIVES: Endografting is being used to manage aneurysms, dissections and acute traumatic disruptions of the thoracic aorta. The acute effects of such interventions on ventricular afterload and on pressure wave transmission characteristics are not well known. METHODS: In five dogs, a 55 mm endograft was introduced into the descending aorta, just distal to the left subclavian artery, with oversizing of 20%. Following formaldehyde induced complete heart block, the hearts were paced (30-120bpm). The ascending aortic pressures and flows were recorded using Millar micro-tip manometers and ultrasonic flowmeters, respectively. Arterial pressures proximal and distal to the stent site were also recorded. For each heart rate, parameters of a modified Windkessel (SVR: systemic vascular resistance, Z0: characteristic impedance, C: total arterial compliance) were estimated. The pulse wave velocity (PWV) and reflection coefficient (Gamma) were calculated from the pressure wave transfer functions. RESULTS: The Z0 (0.25+/-0.05 vs 0.41+/-0.06 mmHg/ml s(-1), P<.05) was increased and C was decreased (0.45+/-0.07 vs 0.28+/-0.04 ml/mmHg, P<0.001) following endograft placement. SVR tended to increase (P=.06) and ascending aortic Gamma was unchanged. The PWV increased (418+/-67 vs 755+/-135 cm/s, P<.05) and the distal Gamma decreased (0.09+/-0.10 vs -0.49+/-0.07, P<.05). CONCLUSIONS: Endografting in the proximal descending aorta cause unfavorable changes in the ascending aortic input impedance and an increase in the PWV through the grafted segment, consistent with an increase in the modulus of elasticity. The grafts produce a negative Gamma at the distal end, an uncommon occurrence in the systemic circulation. Whether this change is of sufficient magnitude to result in post-graft dilation is unknown.

Daphnia magna can tolerate short-term starvation without major changes in lipid metabolism.

Daphnia magna is a common crustacean that is adapted to brief spells of fasting. Lipids are naturally a major component of their diet and are stored as energy reserves. However, there has been some controversy in the literature on the extent to which dietary lipids are used directly for complex lipid formation in Daphnia. We examined lipid metabolism in D. magna by labeling the animals using [1-14C]acetate and then followed the turnover of radiolabeled lipids during a pulse chase. Daphnia were either fed or maintained without food during the chase period. The decrease in radioactivity during the chase was relatively unaffected by feeding, although there were some differences in the distribution of radioactivity between lipid classes or individual FA. The polar lipids, which were four times better labeled than nonpolar lipids, contained the most radioactivity in the zwitterionic phosphoglycerides, PE and PC. Under the experimental conditions, the turnover of the polar membrane lipids was unaffected by feeding. Within nonpolar lipids, TAG accounted for up to about 80% of the label, followed by DAG. Overall, our data show that D. magna is capable of high rates of lipid radiolabeling de novo and, in addition, is able to use--and indeed may be dependent on--some dietary components such as the PUFA linoleate and alpha-linolenate. The results also clearly show that Daphnia is able to tolerate brief spells of fasting (24 h) with very little change to its lipid metabolism.

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).

Investigation into factors affecting precision in ion trap mass spectrometry using different scan directions and axial modulation potential amplitudes.

Electrospray ionization mass spectra obtained from different scan directions are observed to be dependent on the axial modulation potential amplitudes used for resonant ejection and on the positive deviation caused by higher even-multipole fields present in most commercial ion traps. The axial modulation voltage influences the dissociation of ions during resonant ejection and the observed mass shifts. The higher even-multipole fields in commercial ion traps are known to influence resonant ejection from the ion trap and can cause a loss in mass resolution for peaks in reverse scan mass spectra compared with that obtained by the forward scan. However, along with the dissociation of ions during resonant ejection causing a loss in resolution, the possibility of resolving an isotopic distribution is also shown to be influenced by the mass shifts caused by the space charge. These mass shifts differ depending on the scan direction employed. A significant loss in resolution can also result from resonant ejection using non-optimal axial modulation voltages. We also present results showing the ejection of ions at betaz = 1/2 using the reverse scan mode without the axial modulation voltage. Ion ejection at betaz = 1/2 is uncommon in commercial (stretched ion traps) with the conventional analytical scan without the use of a frequency of the axial modulation voltage corresponding to this non-linear resonance.

Investigations into the use of a reverse scan in a quadrupole ion trap mass spectrometer.

The forward scan (i.e. an increasing RF voltage ramp for the mass-selective instability scan) is commonly used as an analytical scan for ion detection with quadrupole ion trap instruments. A number of phenomena have been observed while using this scan technique. These include space charge effects resulting in the delayed ejection of ions from the ion trap, and the fragmentation of fragile ions producing very broad peaks. Here the use of a reverse scan (i.e. a decreasing RF voltage ramp) is examined to determine the effect of the above phenomena in this acquisition method. With regard to space charge effects, the apparent reduction of the carbon isotope spacing below one Thomson (for singly charged ions) that is observed with the forward scan is now replaced by an apparent increase in this spacing. The reverse scan, which optimizes at lower axial modulation ejection voltages than the forward scan, allows for the intact ejection of fragile ions under its typical operating conditions whereas the forward scan results in fragmentation. Reducing the axial modulation voltage for the ejection of ions in the forward scan results in less dissociation of the fragile ions during ion ejection, but with the observation of ghost peaks due to incomplete ejection of all of the ions at the resonance ejection condition. While performing the reverse scan experiment, the formation of product ions from dissociation of the MH(+) ion has also been observed.

Angioedema of the tongue due to acquired C1 esterase inhibitor deficiency.

We describe the management of an 83-year-old woman who presented with upper airway obstruction due to angioedema of the tongue. Following definitive airway management, investigation showed a diagnosis of acquired C1 esterase inhibitor deficiency (acquired angioedema) that was considered to be subsequent to haematological malignancy. Resolution of the macroglossia followed treatment with C1 esterase inhibitor concentrate, but the patient failed to wean from ventilatory support and died in the Intensive Care Unit. This case report highlights the potential for acquired angioedema to cause upper airway obstruction. The various treatment modalities for acquired C1 esterase inhibitor deficiency are summarized.

Genotypic variation in fatty acid content of blackcurrant seeds.

The fatty acid composition and total fatty acid content of seeds from 36 blackcurrant genotypes developed at the Scottish Crop Research Institute were examined. A rapid small-scale procedure, involving homogenization of seeds in toluene followed by sodium methoxide transesterification and gas chromatography, was used. There was considerable variation between genotypes. The gamma-linolenic acid content generally varied from 11 to 19% of the total fatty acids, but three genotypes had higher values of 22-24%, levels previously not reported for blackcurrant seed and similar to those for borage seed. Other nutritionally important fatty acids, stearidonic acid and alpha-linolenic acid, varied from 2 to 4% and 10-19%, respectively. The mean total fatty acid contents ranged from 14 to 23% of the seed, but repeatability was poor. The results are discussed. Blackcurrant seeds are mainly byproducts from juice production, and the study shows the potential for developing blackcurrant genotypes with optimal added value.

Analysis of phospholipid molecular species by liquid chromatography--atmospheric pressure chemical ionisation mass spectrometry of diacylglycerol nicotinates.

A method using liquid chromatography - atmospheric pressure chemical ionisation mass spectrometry was evaluated for determining the molecular species composition of phospholipids (phosphatidylcholines from soybean, egg yolk and bovine liver) after conversion to diacylglycerol nicotinate derivatives. The structures could be deduced from pseudo-molecular ions ([MH-123](+)) and three pairs of monoacyl containing fragment ions. All molecular species in mixed peaks were readily identified and many minor components, earlier not encountered in the samples under investigation, were identified. Acyl chain regioisomers were readily distinguished by the ratio of the [MH-RCHCO](+) ions. Molecular species differing only in the position of the double bonds in one polyunsaturated acyl chain were separated on the basis of retention times. A half quantitative estimation of the molecular species composition of complex samples was achieved by a combination of UV detection and, for mixed peaks, the areas of [MH-123](+) ions.

31P NMR quantitation of phosphorus metabolites in rat heart and skeletal muscle in vivo.

The aim of this study was to examine two methods of 31P NMR quantitation of phosphocreatine (PCr), ATP, and P(i) in rat heart and skeletal muscle in vivo. The first method employed an external standard of phenylphosphonic acid (PPA; 10 mM), and the second method used an enzymatic measurement of tissue ATP equated to the area under the betaATP peak. With the use of the external standard, the concentrations of ATP, PCr, and P(i) in the rat heart were 4.48 +/- 0.33, 9.21 +/- 0.65, and 2.25 +/- 0.16 micromol/g wet wt, respectively. With the use of the internal ATP standard, measured on the same tissue, the contents (means +/- SE) were 4.78 +/- 0.19, 9.83 +/- 0.18, and 2.51 +/- 0.33 micromol/g wet wt, respectively (n = 7). In skeletal muscle, ATP, PCr, and P(i) were 6.09 +/- 0.19, 23.44 +/- 0.88, and 1.81 +/- 0.18 micromol/g wet wt using the PPA standard and 6.03 +/- 0.19, 23.30 +/- 1.30, and 1.82 +/- 0.19 micromol/g wet wt using the internal ATP standard (n = 6). There was no significant difference for each metabolite as measured by the two methods of quantification in heart or skeletal muscle. The results validate the use of an external reference positioned symmetrically above the coil and imply that each has similar NMR sensitivities (similar signal amplitude per mole of 31P between PPA and tissue phosphorus compounds). We conclude that PCr, ATP, and P(i) are nearly 100% visible in the normoxic heart and nonworking skeletal muscle given the errors of measurement.

Negative wave reflections in pulmonary arteries.

The pulmonary arterial branching pattern suggests that the early systolic forward-going compression wave (FCW) might be reflected as a backward-going expansion wave (BEW). Accordingly, in 11 open-chest anesthetized dogs we measured proximal pulmonary arterial pressure and flow (velocity) and evaluated wave reflection using wave-intensity analysis under low-volume, high-volume, high-volume + 20 cmH2O positive end-expiratory pressure (PEEP), and hypoxic conditions. We defined the reflection coefficient R as the ratio of the energy of the reflected wave (BEW [-]; backward-going compression wave, BCW [+]) to that of the incident wave (FCW [+]). We found that R = -0.07 +/- 0.02 under low-volume conditions, which increased in absolute magnitude to -0.20 +/- 0.04 (P < 0.01) under high-volume conditions. The addition of PEEP increased R further to -0.26 +/- 0.02 (P < 0.01). All of these BEWs were reflected from a site ~3 cm downstream. During hypoxia, the BEW was maintained and a BCW appeared (R = +0.09 +/- 0.03) from a closed-end site ~9 cm downstream. The normal pulmonary arterial circulation in the open-chest dog is characterized by negative wave reflection tending to facilitate right ventricular ejection; this reflection increases with increasing blood volume and PEEP.

Force reduction uncoupled from pH and H(2)PO(-)(4) in rat gastrocnemius in vivo with continuous 2-Hz stimulation.

The aim of this study was to examine the effect of the products of ATP hydrolysis on the fatigue process in rat gastrocnemius in vivo. Adult male Sprague-Dawley rats (300-400 g) were anesthetized and ventilated in a custom-built cradle fitted with a force transducer that could be placed into a 7-T NMR magnet. The muscle was stimulated continuously at 2 Hz for 20 min (n = 7). Isometric twitch force increased in the first 4 min of stimulation accompanied by changes in twitch duration (20% increase in relaxation time). Prolonged relaxation was associated with changes in cytosolic pH (6.91 to 6.58), lactate (1.8 to 12.6 micromol/g wet wt), and H(2)PO (7.57 to 13.99 mM). After 4 min, relaxation time, pH, lactate, and H(2)PO returned toward control values as twitch force progressively decreased. No correlation was found between force decline (or twitch broadening) and total phosphate (3 to 23 mM), free [ADP] (18 to 95 microM), free [Mg(2+)] (0.58 to 0.96 mM), or free energy of ATP hydrolysis (-65 to -55 kJ/mol). We conclude that force decline is not due to increased pH and/or H(2)PO but to fatigue of the fast-twitch fibers, possibly linked to glycogen depletion and/or failure of nerve impulse transmission in these fibers.