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Biomedical subjects

R James

Publications and source records attributed to R James.

At least 109 records · Page 6Linked to original sources

Farm and tractor-related fatalities in children in South Australia.

OBJECTIVE: To analyse the circumstances surrounding farm and tractor-related childhood fatalities in South Australia, to identify potentially dangerous situations and to delineate prevention strategies. METHODS: A retrospective search of files was conducted at the Women's and Children's Hospital Department of Histopathology from 1981 to 1996, and the State Coroner's Office from 1988 to 1996. RESULTS: Fifteen fatalities were identified comprising 11 boys and four girls (age range, 2 years 11 months to 13 years; average, 6 years). Activities prior to death included riding on a tractor/trailer (n=8); playing near a field grain storage bin (n=2); playing near machinery (n=2); sleeping in a car near a burn off (n=1); walking around a dam (n=1) and riding as a passenger on a motorbike (n=1). Causes of death included multiple skull fractures and cerebral damage (n=11), asphyxia (n=2), drowning (n=1) and incineration (n=1). CONCLUSIONS: The most dangerous activity involved children riding unrestrained on tractors; falls resulted in extensive injuries from tractor rear wheels or towed machinery. Allowing children on farms to ride on tractors or machinery only if there is a safe seat with a restraining harness would substantially reduce the number of fatal farm-related childhood injuries in South Australia.

Accidents↗

Widespread axonal injury in gunshot wounds to the head using amyloid precursor protein as a marker.

In order to determine whether axonal injury (AI) is a factor in cases of penetrating head injury, the brains of 14 patients who died shortly after sustaining a fatal gunshot wound (GSW) to the head were examined, and the presence of AI determined using immunohistochemical staining for amyloid precursor protein (APP). The distribution of AI was mapped throughout the cerebral hemispheres and brain stem. AI was present in all cases in a diffuse distribution distant to the missle track with severe involvement of the brain stem in all cases. There was no axonal APP immunoreactivity in the direct region of the missle track at the point of primary axotomy. The APP-positive AI in these cases is likely to be a mixture of primary and secondary AI as APP immunostaining is unable to distinguish primary AI due to mechanical deformation from AI secondary to hypoxic-ischemic damage.

Adolescent↗

Interpretations of healthy and unhealthy eating, and implications for dietary change.

Central to many considerations of health-related dietary change is the interpretation given to 'healthy' and 'unhealthy' eating. Results are reported from two studies which investigated people's understanding of these concepts. The first study involved structured interviews with 48 respondents: a wide range of interpretations of healthy and unhealthy eating was found using content analysis techniques. The second study (n = 242) used questionnaires to assess the extent to which different interpretations were endorsed. The extent to which different eating behaviours were perceived as healthy or unhealthy varied greatly. A principal components analysis of perceptions of healthy and unhealthy eating produced five separate factors. Some differences in perceptions were found for people of different age, gender and educational level. Objectively assessed dietary intake and perceived intake were found to be two distinct quantities. 'Healthy' eaters were found to perceive 'healthy eating' slightly differently to 'unhealthy' eaters. The results suggest that lay understanding of healthy eating does generally conform with dietary guidelines, and therefore health promotion priorities should focus on physical and psychological constraints to healthy eating, rather than attempting to increase the public's knowledge as a whole.

Adolescent↗

Amphetamine derivative fatalities in South Australia--is "Ecstasy" the culprit?

OBJECTIVE: To analyze features of a series of fatalities caused by amphetamine-derivative designer drugs marketed as "Ecstasy" in South Australia, and to identify reasons for the recent marked increase in number of these deaths. MATERIALS AND METHODS: Following the death of a 26-year-old woman after alleged ingestion of Ecstasy tablets, a retrospective search of files at State Forensic Science, Adelaide and the South Australian State Coroner's Department was undertaken from February 1992 to January 1997 to identify similar cases. RESULTS: Six fatalities were found, all of which have occurred since September 1995 (M:F ratio, 1:1; age range, 22 to 36 years; average age, 27.7 years). All individuals had histories of recent ingestion of illegal drugs thought to be Ecstasy (methylenedioxymethamphetamine, MDMA) at the time of purchase. Delay occurred in seeking medical attention, despite severe symptoms. Causes of death involved documented hyperthermia in 3 cases (temperatures of 41.5-46.1 degrees C), with features of hyperthermia in one other case, and intracranial hemorrhage in another. Drugs in toxic/lethal amounts identified at postmortem included paramethoxyamphetamine (PMA) in all cases, amphetamine/methamphetamine in 4 cases, and methylenedioxymethamphetamine (MDMA or Ecstasy) in only 2 cases. Interaction with a prescription medication (fluoxetine) may have occurred in 1 case. CONCLUSIONS: The number of deaths due to amphetamine derivatives apparently due to substitution of PMA for MDMA (Ecstasy) have recently increased markedly in Adelaide. Potential users should be warned that PMA has been associated with a much higher rate of lethal complications than other designer drugs, and that no guarantee can be made that tablets sold as Ecstasy are not PMA.

Adult↗

Do shorter-acting neuromuscular blocking drugs or opioids associate with reduced intensive care unit or hospital lengths of stay after coronary artery bypass grafting? CABG Clinical Benchmarking Data Base Participants.

BACKGROUND: The authors hypothesized that shorter-acting opioid and neuromuscular blocking drugs would be associated with reductions in duration of intubation, length of stay (LOS) in the intensive care unit (ICU) after tracheal extubation, or postoperative (exclusive of ICU) LOS, and that shorter durations of intubation would be associated with reduced ICU LOS after extubation and postoperative (exclusive of ICU) LOS. METHODS: One-thousand ninety-four patients undergoing primary coronary artery bypass graft surgery at 40 academic health centers were studied. Multiple patient-related factors were included in multivariate models for hypothesis testing. RESULTS: The duration of tracheal intubation, ICU LOS after extubation, and postoperative (exclusive of ICU) LOS all varied significantly by site. There was no difference between vecuronium and pancuronium in duration of intubation, ICU LOS after extubation, or postoperative (exclusive of ICU) LOS. Use of sufentanil rather than fentanyl was associated with a significant (P=0.045) reduction of 1.9 h (95% CI, 0.04 to 4.1 h) in duration of tracheal intubation but had no significant effect on ICU LOS after extubation, total ICU LOS, postoperative (exclusive of ICU) LOS, or total postoperative LOS. The authors' best model predicts a complex association between increasing duration of intubation and both ICU LOS after tracheal extubation and postoperative (exclusive of ICU) LOS, which was associated with an increase in those measures when duration of intubation exceeded 7.3 or 3 h, respectively. CONCLUSIONS: The LOS measures varied considerably among the institutions. Use of shorter-acting opioid and neuromuscular blocking drugs had no association with ICU LOS after tracheal extubation or with postoperative (exclusive of ICU) LOS. Only when the duration of intubation exceeded threshold values was it associated with increased LOS measures.

Aged↗

Three patients with a 45,X/46,X,psu dic(Xp) karyotype.

Few cases of isochromosomes for the short arm of the X have been reported and all are dicentric with variable portions of the long arms interposed between the two centromeres. This paper reports three cases of complete short arm duplication of one X chromosome in unrelated female patients. All patients also have a 45,X cell line and present with some characteristic features of Turner syndrome. We used conventional cytogenetics, in situ hybridisation, and molecular genetics to describe all three structurally abnormal chromosomes and the parental origin of two of them. We briefly discuss the "inactivation enhancement" theory; however, any genotype-phenotype correlation is complicated by the presence of the 45,X cell line.

Adolescent↗

Phase II study of docetaxel and cisplatin in advanced non-small-cell lung cancer.

PURPOSE: Docetaxel (Taxotere, Rhone-Poulenc Rorer, Antony, France) and cisplatin are two of the most active single agents used in the treatment of non-small-cell lung cancer (NSCLC). A recently reported phase I study of the combination of docetaxel and cisplatin recommended a dose of 75 mg/m2 of both drugs every 3 weeks for subsequent phase II study. PATIENTS AND METHODS: Eligible patients were aged 18 to 75 years with a World Health Organization (WHO) performance status < or = 2 and life expectancy > or = 12 weeks, with metastatic and/or locally advanced NSCLC proven histologically or cytologically. Patients were not permitted to have received prior chemotherapy, extensive radiotherapy, or any radiotherapy to the target lesion and must have had measurable disease. Concurrent treatment with colony-stimulating factors (CSFs) or prophylactic antibiotics was not permitted. Docetaxel (75 mg/m2) in 250 mL 5% dextrose was given intravenously (i.v.) over 1 hour immediately before cisplatin (75 mg/m2) in 500 mL normal saline given i.v. over 1 hour in 3-week cycles. Premedication included ondansetron, dexamethasone, promethazine, and standard hyperhydration with magnesium supplementation. RESULTS: A total of 47 patients, two thirds of whom had metastatic disease, were entered onto this phase II study. The majority of patients were male (72%) and of good (WHO 0 to 1) performance status (85%). All 47 patients were assessable for toxicity and 36 were for response. Three patients were ineligible and eight (17%) discontinued treatment because of significant toxicity. In assessable patients, the overall objective response rate was 38.9% (95% confidence limits [CL], 23.1% to 56.5%), 36.1% had stable disease, and 25% progressive disease. On an intention-to-treat analysis, the objective response rate was 29.8%. Median survival was 9.6 months and estimated 1-year survival was 33%. Significant (grade 3/4) toxicities included nausea (26%), hypotension (15%), diarrhea (13%), and dyspnea mainly related to chest infection (13%). One patient experienced National Cancer Institute (NCI) grade 3 neurosensory toxicity after eight cycles. Grade 3/4 neutropenia was common and occurred in 87% of patients, but thrombocytopenia > or = grade 3 was rare (one patient). Significant (grade 3/4) abnormalities of magnesium levels were common (24%). Febrile neutropenia occurred in 13% of patients and neutropenic infection in 11%, contributing to two treatment-related deaths. No neutropenic enterocolitis or severe fluid retention was reported. CONCLUSION: Compared with other active regimens used in this setting, the combination of docetaxel and cisplatin in advanced NSCLC is an active regimen with a similar toxicity profile to other combination regimens.

Adult↗

Scaling of intrinsic contractile properties and myofibrillar protein composition of fast muscle in the fish myoxocephalus scorpius L

Contractile properties were determined for fast muscle fibres from short-horn sculpin (Myoxocephalus scorpius L.) ranging from 5 to 35 cm total body length (L) and from 2.0 to 758 g body mass. Maximum unloaded shortening velocity (V0) decreased with increasing body size, scaling as 19.5L-0.34, and isometric twitch activation and tetanus relaxation times became longer, scaling as 12.0L0.31 and 19. 5L0.42 respectively. Myofibrillar Mg2+/Ca2+-ATPase activity scaled as 2.51L-0.28. In order to investigate the mechanisms underlying the scaling of contractile properties, myofibrillar protein composition and I filament lengths were determined. One-dimensional SDS-PAGE and two-dimensional isoelectric focusing/non-equilibrium isoelectric focusing-PAGE revealed no differences in the myofibrillar protein isoforms of myosin light chains, actin, tropomyosin, troponin-T and troponin-C in fish of differing body size. Peptide maps of purified myosin heavy chains digested with eight different proteolytic enzymes were also similar in all fish examined. Three isoforms of troponin-I were present in fish less than 20 cm in total length with relative molecular masses of 17 (TnIf3), 22 (TnIf2) and 23 (TnIf1). The ratio of TnI isoforms varied with body length, and only TnIf3 was present in fish greater than 28 cm total length. The length of I filaments was independent of body length. Thus, although the expression of TnI isoforms changes during growth, the underlying mechanism responsible for increased shortening velocity with decreased body size remains unknown.

Journal Article↗

Scaling of muscle performance during escape responses in the fish myoxocephalus scorpius L

Fast-starts associated with escape responses were studied in short-horn sculpin (Myoxocephalus scorpius L.), ranging from 5.5 to 32 cm in total length (L). Electromyography and sonomicrometry were used simultaneously to measure muscle activation and length changes, respectively, in the superficial layers of fast muscle in rostral myotomes. Escape responses consisted of a half tailbeat to bend the body into a C-shape (C-bend), another half tailbeat (contralateral contraction), followed by one or two more tailbeats and/or a gliding phase. The scaling relationships for both muscle strain and shortening duration differed between the C-bend and the contralateral contraction. As a result, relative muscle shortening velocity (V/V0) scaled as -1.18L1.06 for the C-bend and as 1.23L-0. 66 for the contralateral contraction. Therefore, the scaling relationships for muscle shortening velocity varied throughout the time course of the escape response. Muscle power output was determined by using the work-loop technique to subject isolated muscle fibres to in vivo strain and stimulation patterns. Plots of the instantaneous muscle forces and velocities achieved during the contralateral contraction were found to deviate from the steady-state force-velocity relationship. Maximum instantaneous muscle power output was independent of body size, with mean maximum values of 307 and 222 W kg-1 wet muscle mass for the C-bend and the contralateral contraction, respectively.

Journal Article↗

Induction of caspase-3 protease activity and apoptosis by butyrate and trichostatin A (inhibitors of histone deacetylase): dependence on protein synthesis and synergy with a mitochondrial/cytochrome c-dependent pathway.

The induction of apoptosis of tumor cells by the colonic fermentation product butyrate is thought to be an important mechanism in protection against colorectal cancer. Because a major action of butyrate is to inhibit histone deacetylase (leading to chromatin relaxation and altered gene expression), butyrate may induce apoptosis by derepression of specific cell death genes. Here we show that butyrate and trichostatin A (a more selective inhibitor of histone deacetylase) induce the same program of apoptosis in Jurkat lymphoid and LIM 1215 colorectal cancer cell lines that is strictly dependent on new protein synthesis (within 10 h) and that leads to the conversion of the proenzyme form of caspase-3 to the catalytically active effector protease (within 16 h) and apoptotic death (within 24 h). Cells primed with a low concentration of butyrate that itself did not induce activation of caspase-3 or apoptosis were, nevertheless, rendered highly susceptible to induction of apoptosis by staurosporine (an agent that has recently been shown to act by causing mitochondrial release of cytochrome c). Synergy between butyrate and staurosporine was due to the presence of a factor in the cytosol of butyrate-primed cells which enhanced over 7-fold the activation of caspase-3 induced by the addition of cytochrome c and dATP to isolated cytosol. We propose that changes at the level of chromatin structure, induced by a physiological substance butyrate, lead to the expression of a protein that facilitates the pathway by which mitochondria activate caspase-3 and trigger apoptotic death of lymphoid and colorectal cancer cells.

Apoptosis↗

Protein-protein interaction specificity of Im9 for the endonuclease toxin colicin E9 defined by homologue-scanning mutagenesis.

The colicin DNase-specific immunity proteins interact with the endonuclease domain of the bacterial toxin colicin E9 with dissociation constants that span the millimolar to femtomolar affinity range. Among the non-cognate interactions Im2 shows the strongest binding toward the E9 DNase domain with a Kd of 10(-8) M, 6 orders of magnitude weaker than that of the cognate immunity protein Im9. Based on a NMR structure of Im9 that shows it to be a 4-helix protein, we have conducted a mutagenic scan in which elements of Im9 secondary structure were substituted into Im2 to precisely delineate regions that define specificity. Eleven chimeras were constructed, and their biological cross-reactivity toward colicins E2 and E9 was evaluated. From this set of mutants seven proteins were purified, and the Kd for their interaction with the E9 DNase domain was measured by a combination of stopped-flow fluorescence and subunit exchange kinetics. Our results show that immunity specificity is dominated by residues on helix II, accounting for 5 orders of magnitude binding specificity relative to Im2, and that packing interactions of helix II with its neighbor helix I and the loop connecting helix III with helix IV play minor roles. The conformational stability of these chimeric proteins was also determined. Proteins displaying an Im9 phenotype were all more stable than the parent Im2 protein, and surprisingly some chimeras were significantly more stable than either Im2 or Im9.

Amino Acid Sequence↗

Safety of surfaces and equipment for children in playgrounds.

BACKGROUND: The safety of playgrounds is important to protect children from injury, but studies are mostly done mainly under laboratory conditions without epidemiological data. We investigated the safety of different playground surfaces, and types and heights of equipment in public playgrounds in the City of Cardiff, UK. METHODS: We did a correlational study of 330 children aged between 0 and 14 years. All children were hurt when playing in playgrounds in Cardiff and presented to the Accident and Emergency Department in Cardiff Royal Infirmary during summer (April to September) 1992 and 1993, and the whole of 1994. We studied the children's hospital records to establish the type of injury and interviewed their parents to find out the playground and type of equipment involved. The main outcome measures were the number of children injured whilst playing, and injury rates per observed number of children on different surfaces, types, and heights of equipment. FINDINGS: Children sustained significantly more injuries in playgrounds with concrete surfaces than in those with bark or rubberised surfaces (p < 0.001). Playgrounds with rubber surfaces had the lowest rate of injury, with a risk half that of bark and a fifth of that of concrete. Bark surfaces were not significantly more protective against arm fractures than concrete. Most injuries were equipment related. Injury risk due to falls from monkey bars (suspended parallel bars or rings between which children swing) was twice that for climbing-frames and seven times that for swings or slides. The height of the equipment correlated significantly with the number of fractures (p = 0.005) from falls. INTERPRETATION: Rubber or bark surfacing is associated with a low rate of injuries and we support their use in all public playgrounds. Bark alone is insufficient, however, to prevent all injuries, particularly arm fractures. Rubberised impact-absorbing surfaces are safer than bark. We believe that playing on monkeys bars increases the risk of injury in playgrounds and that they should generally not be installed. Safety standards should be based on physical and epidemiological data. Our data suggest that the proposed raising of the maximum fall height from 2.5 m to 3.0 m in Europe is worrying.

Accidental Falls↗

Identification of critical residues in the colicin E9 DNase binding region of the Im9 protein.

1H-15N NMR studies, in conjunction with mutagenesis experiments, have been used to delineate the DNase-binding surface of the colicin E9 inhibitor protein Im9 (where Im stands for immunity protein). Complexes were formed between the 15 kDa unlabelled E9 DNase domain and the 9.5 kDa Im9 protein uniformly labelled with 15N. Approx. 90% of the amide resonances of the bound Im9 were assigned and spectral parameters obtained from 1H-15N heteronuclear single quantum coherence (HSQC) spectra were compared with those for the free Im9 assigned previously. Many of the amide resonances were shifted on complex formation, some by more than 2 p.p.m. in the 15N dimension and more than 0.5 p.p.m. in the 1H dimension. Most of the strongly shifted amides are located on the surfaces of two of the four helices, helix II and helix III. Whereas helix II had already been identified through genetic and biochemical investigations as an important determinant of biological specificity, helix III had not previously been implicated in binding to the DNase. To test the robustness of the NMR-delineated DNase-binding site, a selection of Im9 alanine mutants were constructed and their dissociation rate constants from E9 DNase-immunity protein complexes quantified by radioactive subunit exchange kinetics. Their off-rates correlated well with the NMR perturbation analysis; for example, residues that were highly perturbed in HSQC experiments, such as residues 34 (helix II) and 54 (helix III), had a marked effect on the DNase-immunity protein dissociation rate when replaced by alanine. The NMR and mutagenesis data are consistent with a DNase-binding region on Im9 composed of invariant residues in helix III and variable residues in helix II. The relationship of this binding site model to the wide range of affinities (Kd values in the range 10(-4) to 10(-16)M) that have been measured for cognate and non-cognate colicin DNase-immunity protein interactions is discussed.

Bacterial Proteins↗

[3Fe-4S] <--> [4Fe-4S] cluster interconversion in Desulfovibrio africanus ferredoxin III: properties of an Asp14 --> Cys mutant.

The 8Fe ferredoxin III from Desulfovibrio africanus is a monomeric protein which contains two [4Fe-4S]2+/1+ clusters, one of which is labile and can readily and reversibly lose one Fe under oxidative conditions to yield a [3Fe-4S]1+/0 cluster. This 4Fe cluster has an S = 3/2 ground sping state insteaed of S = 1/2 in the reduced +1 state [George, Armstrong, Hatchikian and Thomson (1989) Biochem. J. 264, 275-284]. The co-ordination to this cluster is unusual in that an aspartate (Asp14, D14, is found where a cysteine residue normally occurs. Using a mutant protein obtained from the overexpression in Escherichia coli of a synthetic gene in which Asp14, the putative ligand to the removable Fe, has been changed to Cys, we have studied the cluster interconversion properties of the labile cluster. Analysis by EPR and magnetic-circular-dichroism spectroscopies showed that the Asp14 --> Cys (D14C) mutant contains two [4Fe-4S]2+/1+ clusters, both with S = 1/2 in the reduced state. Also, unlike in native 8Fe D. africanus ferredoxin III, the 4Fe <--> 3Fe cluster interconversion reaction was found to be sluggish and did not go to completion. It is inferred that the reversibility of the reaction in the native protein is due to the presence of the aspartate residue at position 14 and that this residue might protect the [3Fe-4S] cluster from further degradation.

Amino Acid Sequence↗

New non-peptide endothelin-A receptor antagonists: synthesis, biological properties, and structure-activity relationships of 5-(dimethylamino)-N-pyridyl-,-N-pyrimidinyl-,-N-pyridazinyl-, and -N-pyrazinyl-1-naphthalenesulfonamides.

Use of automated synthesis led to the discovery of several 6-membered nitrogen heterocycles as replacements for the N-isoxazolyl substituent present in the 1-naphthalenesulfonamides endothelin-A (ETA) antagonist 5-(dimethylamino)-N-(3,4-dimethyl-5-isoxazolyl)-1-naphthalenesu lfo namides (BMS 182874). In each of these heterocycles, a small substituent such as halogen para to the position of attachment to the sulfonamide nitrogen atom was found to be advantageous for ETA receptor affinity. Of these heterocycles, 2-pyrazines offered the greatest scope for improving receptor affinity. Optimization of the substituents at the 3- and 5-positions in the pyrazine ring led to potent, ETA-selective compounds such as 5-(dimethylamino)-N-(5-chloro-3-methoxy-2-pyrazinyl)-1- naphthalenesulfonamides (7m, ETA pIC50 8.1). When dosed orally at 10 mg/kg to conscious, normotensive rats infused with big ET-1, compounds such as 7m showed significant inhibition of the pressor response with a duration of effect lasting for the 5-h course of the experiment.

Animals↗

Homeosis and intestinal tumours in Cdx2 mutant mice.

In Drosophila, disturbing the expression of the homeobox gene caudal causes a severe disruption in body segmentation and global body patterning. There are three mouse homologues of Drosophila caudal: Cdx1 (ref. 2), Cdx2 (ref. 3) and Cdx4 (ref. 4). We have generated a null mutation of murine Cdx2 by homologous recombination. Cdx2 homozygote null mutants die between 3.5 and 5.5 days post coitum (d.p.c.). Cdx2 heterozygote mutants exhibit a variable phenotype, with many showing tail abnormalities or stunted growth. Skeletal analysis demonstrates a homeotic shift of vertebrae and compatible malformations of the ribs. Within the first three months of life, 90% of Cdx2 heterozygotes develop multiple intestinal adenomatous polyps, particularly in the proximal colon. These polyps occasionally contain areas of true metaplasia. In contrast to the surrounding intestinal epithelium, the neoplastic cells do not express Cdx2 from the remaining allele. These results suggest that Cdx2 mutation is the primary event in the genesis of some intestinal tumours.

Adenomatous Polyposis Coli↗

Determination of the pesticide fenbutatin oxide in tomatoes, cucumbers and bananas by high performance liquid chromatographic/atmospheric pressure chemical ionization-mass spectrometry.

A rapid method, using high-performance liquid chromatography (HPLC)/ atmospheric pressure chemical ionization-mass spectrometry, has been developed for the determination of fenbutatin oxide in tomatoes, cucumbers and bananas. Samples were homogenized with sodium carbonate and ethyl acetate, filtered through sodium sulphate, concentrated and solvent exchanged into acetonitrile prior to analysis. HPLC was performed on a Hypercarb column with 10:90 acetic acid (5% v/v glacial acetic acid in water)/acetonitrile at a flow rate of 1 mL/min. Positive ionization selected-ion monitoring was performed on the 7 isotopic cluster ions from the tris(2-methyl-2-phenylpropyl) tin fragment. A comparison of solvent-based and extract-based standards showed that tomato and cucumber matrices had a slight enhancement effect on the signal intensity, whereas the banana matrix exerted a signal suppression effect. Calibration was linear over the range 0.25-5.0 ng/microL. The mean spike recoveries (extracts spiked at 0.5 mg/kg) were 88% for tomatoes and 80% for both cucumbers and bananas with relative standard deviations of 6%, 7% and 8% respectively. Limits of detection were commodity dependent and ranged from 0.06-0.12 ng/microL (equivalent to 0.01-0.02 mg/kg in the crop). Ionization was stable for long analytical time periods.

Chromatography, High Pressure Liquid↗