Medical provision in forward locations in Afghanistan: the experiences of General Duties Medical Officers on Op HERRICK 15.

Op HERRICK in Afghanistan remains the current focus of the British Armed Forces. General Duties Medical Officers (GDMOs) commonly deploy to Role 1 locations in Afghanistan, which remains a continuously evolving theatre of operations. This article is based on the experiences of four GDMOs who deployed to forward Company locations on Op HERRICK 15 (September 2011-April 2012) and aims to offer an insight into the challenges of this role and identify areas in which improvements could be made to the training and preparation for future tours.

Exploring the limits of bacterial identification by intact cell-mass spectrometry.

The limits of intact cell-mass spectrometry (ICM-MS) were tested with regard to the minimum number of bacterial cells detectable and its power to discriminate mixed-bacterial cultures. The technique is a surface analysis tool, as is supported by evidence showing that mass fingerprints correspond to material desorbed directly from the cell wall. The brief exposure to solvents, which occurs during sample preparation, does not extract internal cellular material. Spectra were collected over the m/z range of 500 to 10,000. The UV absorbing matrices used were found to be highly specific to bacterial gram type: alpha-cyano-4-hydroxycinnamic acid for gram-negative bacteria and 5-chloro-2-mercaptobenzothiazole for gram-positive bacteria. This specificity allows mixed cultures of different gram types to be differentiated by ICM-MS. The minimum number of cells that could reliably give spectra of sufficient data was 10(4) cells (10(7) cells/mL).

Phospholipid analogue profiles of Peptostreptococcus, Micromonas, and Finegoldia species analysed by fast atom bombardment mass spectrometry.

Species of Peptostreptococcus cause a variety of infections, primarily abscesses of soft tissues, joints, and mucous membranes. The aim of this study was to compare the phospholipid analogue profiles of Peptostreptococcus species, represented by P. anaerobius, P. asaccharolyticus, P. indolicus, P. lacrimalis, and P. prevotii; Micromonas micros (P. micros) and Finegoldia magna (P. magnus). After anaerobic growth on blood-FAA, lipids extracted by chloroform-methanol (2:1 v/v) were purified, then analysed by fast atom bombardment mass spectrometry (FAB-MS) in negative ion mode. The major peaks with mass to charge (m/z) 719, 721, and 749, corresponded to phosphatidylglycerol analogues, namely PG (32:1), PG (32:0), and PG (34:0), which have been found previously in Lactobacillus spp., Clostridium difficile, and Staphylococcus spp. Other major peaks observed, with m/z 619, 647, 665, 675, 677, 687, 691, 693, 701, 703, 707, 733, and 746 have also been reported in one or more of these three species. However, other major peaks found here in Peptostreptococcus, Micromonas, and Finegoldia have not been described elsewhere; these are 501, 514, 515, 618, 659, 673, 676, 688, 690, 692, 694, 700, 706, 715, 718, 722, and 750. We conclude that Peptostreptococcus, Micromonas, and Finegoldia isolates are chemically unique.

Effects of ion mode and matrix additives in the identification of bacteria by intact cell mass spectrometry.

Protocols for the identification of bacterial cells by intact cell matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (ICM-TOFMS) are presented. A mass range of 500 to 10,000 m/z is used. The use of formic acid and the crown ether 1, 4, 7, 10, 13, 16-hexaoxacyclooctadecane (18-crown-6) is described. Crown ether is useful for removing metal ion adducts, which degrade spectral purity, and formic acid promotes positive ions, improves spectral signal, and, hence, increases identification certainty.

Rapid discrimination between methicillin-sensitive and methicillin-resistant Staphylococcus aureus by intact cell mass spectrometry.

Rapid, accurate discrimination between methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) strains is essential for appropriate therapeutic management and timely intervention for infection control. A rapid method involving intact cell mass spectrometry (ICMS) is presented that shows promise for identification, discrimination of MSSA from MRSA and typing. In ICMS, cells from a bacterial colony are emulsified in a chemical matrix, added to a sample slide, dried and analysed by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS). This technique examines the chemistry of the intact bacterial cell surface, yielding spectra consisting of a series of peaks from 500 to 10000, which represent the mass:charge (m:z) ratios. Each peak corresponds to a molecular fragment released from the cell surface during laser desorption. Specimens can be prepared in a few seconds from plate cultures and a spectrum can be obtained within 2 min. ICMS spectra for 20 staphylococcal isolates showed characteristic peaks, some of which were conserved at species level, some at strain level and some were characteristic of the methicillin susceptibility status of the strain. ICMS may have potential for MRSA identification and typing, and may improve infection control measures.

The rapid identification of intact microorganisms using mass spectrometry.

Antibiotic-resistant strains of bacteria continue to emerge, increasing the need for their fast and accurate identification. Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), has become a prominent technique in biological mass spectrometry. We report the application of MALDI-TOF-MS for the identification of intact Gram-negative and Gram-positive microorganisms taken directly from culture. Analysis of bacteria from a single colony is possible, allowing the screening of mixed cultures. Sample preparation is simple and the analysis automated, providing spectra within minutes. The spectra obtained allow identification of microorganisms from different genera, different species, and from different strains of the same species. The procedure provides a unique mass spectral fingerprint of the microorganism, produced from desorbed components of the cell wall. Consistent data were obtained from subcultures grown for 3-day and 6-day periods, from the same cultures 1 day later and from fresh subcultures 2 months later.

Twenty-four hour intermittent perfusion storage of the isolated rat heart: the effect of perfusion intervals on functional preservation.

We investigated the effect of intermittent perfusion intervals on heart preservation. The isolated rat heart was flushed with a cardioplegic solution, CP-11EB, and stored at 0 degrees C for 24 hours. During storage, the heart was perfused periodically at 60 mm Hg for 3 minutes with 25 degrees C oxygenated CP-11EB. The perfusion schedules were as follows: group I, every 4 hours; group II, every 6 hours; group III, every 8 hours; group IV, every 10 hours; group V, 10 and 17 hours of storage; group VI, every 11 hours. Poststorage function was assessed after 30 minutes of working reperfusion. Function of the unstored hearts including aortic flow (51.5 +/- 2.5 ml/min), coronary flow (24.5 +/- 1.3 ml/min), cardiac output (75.6 +/- 3.0 ml/min) and work (85.0 +/- 5.4 g-ml/min) served as controls. Group V hearts were best preserved with aortic flow recovered to 65%; coronary flow 44%; cardiac output 58%; and work 53% of the control. Recovery in group IV was comparable with group V. Groups I, II, III, and VI recovered significantly less than group V. Myocardial adenosine triphosphate content (micromoles per gram dry) in group V was 23.8 +/- 1.7, 20.8 +/- 0.9, 13.5 +/- 2.4, and 15.0 +/- 4.0, at 0, 10, 17, and 24 hours of storage, respectively. Intermittent perfusion at 10 and 17 hours elevated adenosine triphosphate to 99% and 88% of prestorage level. Poststorage reperfusion did not improve improve adenosine triphosphate content (16.2 +/- 2.4) over the end-storage level.(ABSTRACT TRUNCATED AT 250 WORDS)

Enhanced functional preservation of cold-stored rat heart by a nucleoside transport inhibitor.

This study investigates the hypothesis that inhibition of nucleoside transport during hypothermic storage elevates tissue adenosine (ADO) content and improves the function of the isolated rat heart. The hearts, flushed with a cardioplegic solution containing varying concentrations (0-100 nM) of a nucleoside transport inhibitor, S-(4-nitrobenzyl)-6-thioinosine (NBTI), were immersion-stored at 0 degrees C for 9 hr. Function was assessed after 30 min of working reperfusion. Function of unstored fresh hearts served as controls and poststorage recovery is reported as percentage of control function. Poststorage heart rate in all groups returned to control level after reperfusion. Recovery of other functional parameters in the no-NBTI group was as follows: aortic flow (AF), 56.2 +/- 4.6%; coronary flow (CF), 53.9 +/- 3.2%; cardiac output (CO), 55.5 +/- 4.0%; systolic pressure, 81.6 +/- 2.5%; work, 47.0 +/- 4.2%; and coronary vascular resistance (CVR), 157.1 +/- 7.8% of control. NBTI improved functional recovery in a dose-dependent fashion; the maximal improvement was seen at a dose of 5 nM, in which the recovery was: AF, 78.1 +/- 3.4%; CF, 73.5 +/- 4.4%; CO, 76.7 +/- 3.6%; work, 70.7 +/- 5.0%; and CVR, 127.5 +/- 4.5% of control (P < 0.05 vs. no-NBTI). The ADO A1-receptor antagonist, 1,3-dipropyl-8-cyclopentylxanthine (0.1 microM) blocked the effects of 5 nM NBTI; the recovery of AF, CF, CO, work, and CVR decreased to 62.8 +/- 8.0%, 58.3 +/- 5.0%, 61.5 +/- 3.9%, 54.4 +/- 4.5%, and 163.8 +/- 12.7% of control, respectively (P < 0.05 vs. 5 nM NBTI). Tissue ADO content in 5 nM NBTI hearts at the end of storage was 0.075 +/- 0.025 mumol/g dry wt, which was significantly elevated from 0.016 +/- 0.004 mumol/g dry wt in no-NBTI hearts. Purine release during initial reperfusion was delayed in 5 nM NBTI hearts, indicating the inhibition of nucleoside transport by NBTI. But NBTI treatment did not improve end-storage or end-reperfusion myocardial ATP. In conclusion, the addition of NBTI to cardioplegic solution enhanced tissue ADO and improved poststorage function of the hypothermically stored rat heart. The effect is ADO A1-receptor mediated without invoking energy conservation.

Adenosine deaminase inhibitor in cardioplegia enhanced function preservation of the hypothermically stored rat heart.

Adenosine (ADO) has been shown to be protective to the ischemic-reperfused myocardium. This study tested the hypothesis that inhibition of myocardial adenosine deaminase during cold storage will elevate tissue ADO content, improve the cardiac function, and preserve ATP. The isolated rat hearts (6-9 hearts/group) were flushed with a cardioplegic solution containing 0-75 microM erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and immersion-stored at 0 degree C for 9 hr. Function was assessed after 30 min working reperfusion. Function of the unstored hearts (n = 11, mean +/- SE) including heart rate (293 +/- 13 bpm), aortic flow (AF; 52.5 +/- 1.1 ml/min), coronary flow (CF; 23.5 +/- 1.3 ml/min), cardiac output (CO; 76.0 +/- 2.1 ml/min), systolic pressure (SP; 136 +/- 2 mmHg), diastolic pressure (DP; 63 +/- 1 mm Hg), work (90.5 +/- 3.4 g-m/min), and coronary vascular resistance (CVR; 2.77 +/- 0.14 mmHg-min/ml) served as controls. Heart rate in all stored hearts returned to normal after reperfusion. Recovery of other function in no-EHNA group was: AF, 52 +/- 7; CF, 55 +/- 5; CO, 53 +/- 6; SP, 79 +/- 4; DP, 93 +/- 3; work, 47 +/- 7; and CVR, 171 +/- 15% of control. EHNA improved functional recovery in a dose-dependent fashion. At the optimal concentration of 25 microM, the recovery was: AF, 83 +/- 6; CF, 68 +/- 4; CO, 78 +/- 5; SP, 90 +/- 3; DP, 105 +/- 5; work, 77 +/- 8; and CVR 151 +/- 9% of control. ADO A1 receptor antagonists, 8-phenyltheophylline (1 microM) and 1,3-dipropyl-8-cyclopentylxanthine (0.1 microM) blocked the effects of 25 microM EHNA; the recovery of CO was reduced to 65 +/- 3 and 50 +/- 2% of the control, respectively. Tissue ADO content in 25 microM EHNA hearts at the end of storage was 95 +/- 19 nmol/g dry wt, which was significantly elevated from 15 +/- 3 nmol/g dry wt in no-EHNA hearts. EHNA also caused a 45-fold increase in the release of ADO over no-EHNA group during the first 10 min of reperfusion. But EHNA treatment did not cause any change in either end-storage or end-reperfusion myocardial ATP levels. Thus EHNA in cardioplegic solution inhibited cardiac ADO catabolism during long-term hypothermic storage and improved function preservation partially via an ADO A1 receptor-mediated mechanism without invoking ATP conservation.