Serum Organophosphorus compound levels as a surrogate marker for severity assessment and management in Acute Organophosphorus Poisoning - A novel approach.

Organophosphorus (OP) compounds are the most extensively used pesticides' class worldwide; cause most self­poisoning deaths especially in India. Thus, it is utmost important for early identification and aggressive management of OP poisoning from the clinical perspective to prevent serious complications by using sophisticated LC-MS/MS approach. This was a prospective study involving 103 patients of OP cases admitted to Karnataka Institute of Medical Sciences from June 2022 to May 2023, based on the inclusion and exclusion criteria patients were subjected to study. On admission, venous blood was collected from patient with Malathion and Profenofos OP poisoning history and subjected to serum biomarker and to LC-MS/MS analysis. Out of the 103 patients, 68 patients consumed Profenofos (66%) and 35 patients consumed Malathion (34%). Pseudocholinesterase levels among the of OP cases revealed that the 33 patients had mild toxicity, 40 patients had moderate toxicity and 30 patients had severe toxicity of OP poisoning. Subsequently LC-MS/MS analysis showed that the results obtained are not in correlation with indirect serum marker pseudocholinesterase levels. On the other side, LC-MS/MS results are in correlation with the clinical outcome of the patients with respect to morbidity and mortality. Thus, LC-MS/MS approach to assess the OP levels in patients could be used as potential diagnostic and prognostic marker for the absolute quantification of OP compounds compared to indirect OP levels estimation.

Expression of Hypersensitive Troponin I and Soluble ST2 in Acute Organophosphorus Pesticide Poisoning.

The role of soluble growth stimulating gene 2 protein and highly sensitive cardiac troponin in the diagnosis of early myocardial injury caused by acute organophosphorus pesticide poisoning was studied. 171 inpatients with AOPP were divided into three experimental groups according to their mild, moderate, and severe conditions. 20 healthy people were selected as the control group. The levels of cTnI, HS-CTNI, NT proBNP, and ST2 were measured at the 4th and 12th hours after the experiment. The measured data were expressed by mean standard deviation. The independent sample t-test was used for the detection between the two groups, and one-way ANOVA was used for the analysis and comparison between multiple groups. The relevant data were analyzed by Spearman correlation test (P < 0.05). The levels of cTnI and HS cTnI in the experimental group increased with the extension of time and the deepening of poisoning degree; four hours after admission, ST2 and NT proBNP water in the control group and the experimental group increased significantly on average. According to the analysis of the data, there was a positive correlation between HS TnI and ST2 in patients with AOPP (r = 0.938, P < 0.001, r = 0.827, P < 0.001). The more serious the disease, the higher the concentrations of HS TnI and ST2, and the more serious the myocardial injury.

Application of Q-TOF-MS based metabonomics techniques to analyze the plasma metabolic profile changes on rats following death due to acute intoxication of phorate.

Organophosphorus pesticides (OPS) are widely used in the world, and many poisoning cases were caused by them. Phorate intoxication is especially common in China. However, there are currently few methods for discriminating phorate poisoning death from phorate exposure after death and interpretation of false-positive results due to the lack of effective biomarkers. In this study, we investigated the metabonomics of rat plasma at different dose levels of acute phorate intoxication using ultra-performance liquid chromatography quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS) analysis. A total of 11 endogenous metabolites were significantly changed in the groups exposed to phorate at LD50 level and three times of LD50 (3LD50) level compared with the control group, which could be potential biomarkers of acute phorate intoxication. Plasma metabonomics analysis showed that diethylthiophosphate (DETP) could be a useful biomarker of acute phorate intoxication. The levels of uric acid, acylcarnitine, succinate, gluconic acid, and phosphatidylcholine (PC) (36:2) were increased, while pyruvate level was decreased in all groups exposed to phorate. The levels of ceramides (Cer) (d 18:0/16:0), palmitic acid, and lysophosphatidylcholine (lysoPC) (18:1) were only changed after 3LD50 dosage. The results of this study indicate that the dose-dependent relationship exists between metabolomic profile change and toxicities associated with apoptosis, fatty acid metabolism disorder, energy metabolism disorder especially tricarboxylic acid (TCA) cycle, as well as liver, kidney, and nervous system functions after acute exposure of phorate. This study shows that metabonomics is a useful tool in identifying biomarkers for the forensic toxicology study of phorate poisoning.

The clinical role of serum albumin in Organophospate poisoning.

This retrospective study investigated whether the serum albumin (SA) concentration at presentation is associated with mortality and the mechanism underlying the association. This study enrolled 217 patients poisoned with organophosphate (OP). Hypoalbuminemia (albumin <3.5 g/dL) at presentation was identified in 18.4% of the patients poisoned with OP. The hypoalbuminemia group experienced a more complicated clinical course and had a higher mortality rate than the normoalbuminemia and hyperalbuminemia groups. The SA concentration correlated with the CRP level at presentation but not with the body mass index in patients with OP poisoning. Furthermore, the change in the SA concentration during the first 24 hours also correlated with the change in BuChE activity in patients with fenitrothion poisoning. The SA concentration at presentation was independently associated with mortality after adjusting for inflammation and nutritional status. This study showed that the SA concentration at presentation is associated with the mortality risk of patients poisoned with OP. This association is independent of inflammation and nutritional status in OP poisoning, and in particular, the protective effect of SA might contribute to this association in fenitrothion poisoning. These results should be validated.

A case report of cholinesterase inhibitor poisoning: cholinesterase activities and analytical methods for diagnosis and clinical decision making.

Suicidal ingestion of organophosphorus (OP) or carbamate (CM) compounds challenges health care systems worldwide, particularly in Southeast Asia. The diagnosis and treatment of OP or CM poisoning is traditionally based on the clinical appearance of the typical cholinergic toxidrome, e.g. miosis, salivation and bradycardia. Yet, clinical signs might be inconclusive or even misleading. A current case report highlights the importance of enzymatic assays to provide rapid information and support clinicians in diagnosis and rational clinical decision making. Furthermore, the differentiation between OP and CM poisoning seems important, as an oxime therapy will most probably not provide benefit in CM poisoning, but-as every pharmaceutical product-it might result in adverse effects. The early identification of the causing agent and the amount taken up in the body are helpful in planning of the therapeutic regimen including experimental strategies, e.g. the use of human blood products to facilitate scavenging of the toxic agent. Furthermore, the analysis of biotransformation products and antidote levels provides additional insights into the pathophysiology of OP or CM poisoning. In conclusion, cholinesterase activities and modern analytical methods help to provide a more effective treatment and a thorough understanding of individual cases of OP or CM poisoning.

Toxicity Evaluation of the Subacute Diazinon in Aged Male Rats: Hematological Aspects.

BACKGROUND & OBJECTIVE: Age-dependent Organophosphates (OPs) toxicity is a controversial topic. The present study was designed to investigate the effect of the sub-acute exposure to diazinon (DZN), one of the main OPs insecticides, on the hematological alterations in adult and aged male rats. METHODS: For the aim of this approach, the adult and aged rats were administered with DZN (15 mg/kg, orally) for 4 weeks. Then, the blood samples were collected from the retro-orbital sinus for measuring red blood cell (RBC), hemoglobin (Hb), hematocrit (Hct), platelets (PLT), MCV (mean corpuscular volume), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin (MCHC). RESULTS: The obtained results indicated that DZN significantly decreased RBCs (4.93 ± 0.41), Htc (28.12 ± 1.21), Hb (10.31 ± 0.36), MCHC (30.51 ± 2.04), MCV (62.86 ± 2.58), and PLT (265.6 ± 34.81) values in the adult and aged rats versus the age-matched control rats. Moreover, RBC, Hb, and Htc levels decreased significantly in the aged rats versus adult rats. However, no significant differences were observed between MCHC, MCV, and PLT levels in adult and aged rats. Moreover, the MCH concentration did not change in any group. Additionally, DZN did not deteriorate the hematological alterations in the aged rats versus adult rats. CONCLUSION: the present study showed that the toxicity of DZN is not associated with age. However, more studies should be conducted to confirm this finding.

Serum ubiquitin C-terminal hydrolase L1 predicts cognitive impairment in patients with acute organophosphorus pesticide poisoning.

BACKGROUND: To assess the usefulness of serum C-terminal hydrolase L1 (UCH-L1) level as a biomarker for predicting cognitive impairment in patients with acute organophosphorus pesticide poisoning (AOPP). METHODS: Two hundred and seven adult patients with AOPP were included in this study. Serum UCH-L1 levels were assessed on admission (Day 1 postpoisoning) and on Days 3 and 7 postpoisoning. The associations between serum UCH-L1 levels, other clinical predictors, and cognitive function evaluated on Day 30 postpoisoning were investigated. RESULTS: On multivariate analysis, serum UCH-L1 levels on admission (odds ratio [OR] 1.889, 95% confidence interval [CI] 1.609-3.082, P = 0.002) and 24-hour APACHE II score (OR 1.736, 95% CI 1.264-3.272, P = 0.012) were independent predictors of cognitive impairment on Day 30 postpoisoning. Based on the receiver operating characteristic curve, serum UCH-L1 levels >5.9 ng/mL on admission predicted cognitive impairment on Day 30 postpoisoning with 86.1% sensitivity and 72.5% specificity (area under the curve, 0.869; 95% CI 0.815-0.923). On admission [8.51 (6.53-10.22) ng/mL vs 4.25 (2.57-6.31) ng/mL, P < 0.001] and Day 3 [9.31 (7.92-10.98) ng/mL vs 3.32 (2.25-5.13) ng/mL, P < 0.001] and Day 7 [4.96 (3.28-7.26) ng/mL vs 2.27 (1.55-3.24) ng/mL, P < 0.001] postpoisoning, serum UCH-L1 concentration was significantly higher in patients that developed cognitive impairment compared to those that did not. CONCLUSION: This study demonstrates that serum UCH-L1 level has potential as a novel biomarker for predicting cognitive impairment 30 days after AOPP.

Platelet-lymphocyte ratio is not a prognostic predictor for acute paraquat-intoxicated patients: A retrospective analysis.

This study aimed to investigate the prognostic predictive value of the platelet-lymphocyte ratio (PLR) in patients with acute paraquat (PQ) intoxication.A total of 107 patients with acute PQ intoxication via oral ingestion were admitted in Cangzhou Central Hospital from May 2012 to September 2018. Valuable detection indices were screened out by using Cox proportional hazard regression and receiver operating characteristic (ROC) curve analyses, and their diagnostic efficiency was evaluated by using Kaplan-Meier curve.The 90-day mortality was 58.9% (63/107). The Kaplan-Meier curve showed that PLR was not associated with 90-day survival (log-rank test; P = .661). In Cox proportional hazard regression analyses, PLR was not an independent risk factor. Meanwhile, the ROC curves showed that PLR had an AUC value of 0.569 (95% confidence interval: 0.459-0.679, P = .227) in predicting 90-day survival.PLR is not a prognostic predictor for patients with acute PQ intoxication.

Manifestations of and risk factors for acute myocardial injury after acute organophosphorus pesticide poisoning.

This study aimed to explore the risk factors for acute myocardial injury (AMI) caused by acute organophosphorus pesticide poisoning (AOPP).The clinical data of 98 patients, who were treated in our hospital due to oral AOPP from April 2013 to April 2017, were retrospectively analyzed. These patients were divided into two groups: AMI group and control group. The incidence of AMI was analyzed. Furthermore, the dosage forms and dose of the pesticide, and the interval between pesticide taking and doctor visit were compared between these two groups. Moreover, their clinical symptoms were observed; the serum cholinesterase levels, myocardial injury, and heart failure markers were detected, and the occurrence of arrhythmia and the structure and function of the heart were investigated through continuous electrocardiographic monitoring and transthoracic echocardiography.Among these 98 AOPP patients, 51 patients were complicated with AMI, and the incidence was 52.0%. The main manifestations of these 51 patients with AMI were as follows: the serum levels of myocardial injury markers (creatine kinase-Mb [CK-Mb] and cardiac troponin I [cTnI]) and heart failure markers (N-terminal pro B-type natriuretic peptide [NT-pro BNP]) were significantly higher, when compared with the control group (P < .001), and the incidence of arrhythmia (FVPB, P = .02; RAA, P = .03; RVA, P = .02; ST-T changes, P = .01) and heart failure (P = .04) was also significantly higher when compared with the control group. With regard to dosage forms of the pesticides, the number of patients taking the pesticides with solvents containing aromatic hydrocarbons was significantly higher in the AMI group than in the control group (P = .001). And the number of patients taking over 100 mL of pesticides was also significantly higher in the AMI group than in the control group (P < .001). Significantly more patients in the AMI group had an interval of over 1 h between pesticide taking and doctor visit than in the control group (P < .001).Risk factors for AMI after AOPP may include the dose and dosage form of the pesticide, and the interval between pesticide taking and doctor visit.

Evaluation of the accuracy of "ChE check mobile" in measurement of acetylcholinesterase in pesticide poisoning.

BACKGROUND: Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are used in clinical management to confirm the diagnosis and indicate the severity of organophosphorus and carbamate poisoning. ChE check mobile is a new portable cholinesterase testing system developed in Germany. The study aims to evaluate the accuracy of ChE check mobile compared to the standard reference method and Test-mate ChE system. METHODS: Patients with organophosphorus and carbamate poisoning were recruited from two general hospitals in Sri Lanka between September 2013 and November 2014. The AChE was measured using the three methods. RESULTS: Blood samples were collected from 185 self-poisoned patients (170 organophosphorus and 15 carbamate) and 20 normal individuals. ChE check mobile correlated well with spectrophotometer readings (Pearson's correlation coefficient 0.87) but gave higher values (Mean bias for AChE: +6.55 (95% CI: -11 to 24) U/g Hb). A similar positive bias from Test-mate results was also observed. Applying a correction factor derived from the volunteer samples (dividing by 1.353) greatly improved agreement in pesticide poisoned patients. CONCLUSIONS: ChE check mobile system allowed for rapid determination of AChE activity but gave somewhat higher AChE compared to other methods. Applying a correction factor of 1.353 provide a good agreement to both reference and Test-mate ChE machine in this setting.

An experimental study on investigating the postmortem interval in dichlorvos poisoned rats by GC/MS-based metabolomics.

The estimation of the postmortem interval (PMI) is always a key issue in forensic science. Although many attempts based on metabolomics approaches have been proven to be feasible and accurate for PMI estimation, there have been no reports regarding the determination of the PMI in acute dichlorvos (DDVP) poisoning. In this study, all rats were killed by acute DDVP poisoning at a dose three fold the oral LD50 (240 mg/kg). Gas chromatography-mass spectrometry (GC/MS) was applied to investigate the metabolic profiling of blood samples at various times after death up to 72 h. A total of 39 metabolites were found to be associated with PMI, and the combinations of various numbers of metabolites were used to establish support vector regression (SVR) models to investigate the PMI. The SVR model constructed by 23 metabolites had a minimum mean squared error (MSE) of 5.49 h for the training set. Then, the SVR model was validated by prediction set with an MSE of 10.33 h, suggesting good predictive ability of the model for investigating the PMI. The findings demonstrated the great potential of GC/MS-based metabolomics combined with the SVR model in determining the PMI of DDVP poisoned rats and provided an experimental basis for the application of this approach in investigating the PMI of other toxicants.

Lipid emulsion for acute organophosphate insecticide poisoning - a pilot observational safety study.

INTRODUCTION: Organophosphate (OP) poisoning is an important public health concern in South-Asia especially in the farming population. Treatment of OP poisoning has remained unchanged since decades and case fatality is 10-40% despite best supportive care, anticholinergic agents and oximes. A new antidote is the need of the hour. Lipid emulsion being inexpensive, easily available and effective in management of other lipid soluble toxins may be a novel option. However, safety has not been established and efficacy is limited to animal studies and case reports in humans with OP poisoning. METHODS: An open-label pilot study was undertaken to establish the safety of lipid emulsion in OP poisoning. Patients with symptomatic OP poisoning, meeting the inclusion and exclusion criteria were treated with 100 mL of 20% intravenous lipid emulsion (after consent) in addition to standard of care. They were monitored for change in hemodynamic parameters, change in hematology and biochemical parameters at various intervals of time after initiation of therapy. Morbidity, mortality, and occurrence of adverse effects were compared with historical control group. RESULTS: Forty patients with symptomatic OP poisoning were enrolled in the study group. No significant change in hemodynamic parameters (pulse rate, systolic, diastolic blood pressure, and mean arterial pressure) or in hematology and biochemical parameters were seen. No adverse effects were noted. Compared to historical controls, no change in mortality was noted, although there was reduced duration of mechanical ventilation, hospital stay, and early resolution of hypernatremia. CONCLUSIONS: This study evaluates the safety of lipid emulsion in OP poisoning. Absence of change in hemodynamic parameters and adverse effects suggests lipid emulsion may be safe for this indication. Large randomized controlled trials are now required to assess clinical efficacy. ClinicalTrials.gov number: NCT03564574.

Análise do impacto do uso de organofosforados e carbamatos em trabalhadores rurais de um município da região noroeste do estado do Rio Grande do Sul / Impact analysis of the use of organophosphates and carbamates rural workers of the municipality in a northwest region of Rio Grande do Sul

Anualmente milhões de agricultores são intoxicados no mundo, e destes, mais de 20 mil morrem em consequência da exposição a agrotóxicos. Intoxicações por organofosforados (OF) e carbamatos (CAR) representam as maiores ameaças à saúde dos trabalhadores rurais. Os OF e CAR atuam na inibição da enzima colinesterase, sendo assim a inibição desta mostra-se um excelente indicador da severidade da intoxicação. O objetivo deste estudo foi analisar o impacto do uso de OF e CAR em trabalhadores rurais na cidade de Mato Queimado/RS. Foi realizado um estudo transversal, prospectivo e experimental. Investigaramse 27 trabalhadores rurais expostos. Foram realizadas coletas sanguíneas e dados epidemiográficos nos meses de fevereiro e março de 2014. A atividade da colinesterase foi determinada através do método bioquímico cinético colorimétrico. A faixa etária média dos participantes foi 34,6 anos (± 8,5). A forma de contato mais prevalente foi a aplicação do produto (88,9%). O tempo médio de exposição foi de 10,7 anos. 70,4% relataram usar equipamentos de proteção individual (EPI), sendo mais frequente o uso de máscara (55,5%). A média dos valores de colinesterase para foi de 3244,45 U/I (± 345,8), níveis estes abaixo dos de referência. Através dos resultados obtidos nesta pesquisa torna-se imprescindível a utilização de meios de monitoramento biológico dos trabalhadores rurais na finalidade de prevenção e promoção da saúde.

Annually millions of rural workers are intoxicated in the world, and of these, more than 20,000 die as a result of exposure to pesticides. Intoxication by insecticides organophosphate (OF) and carbamates (CAR) represent the greatest threats to the health of rural workers. OF CAR and act on the inhibition of cholinesterase enzyme, thus inhibition of this proves to be an excellent indicator of the severity of the intoxication. The objective of this study was to analyze the impact of using OF CAR and in rural workers in the city of Mato Queimado/RS. A cross-sectional, prospective and experimental study was conducted. Twenty-three rural workers exposed were investigated. Sample collection and data demographic were conducted in February and March 2014. The cholinesterase activity was determined by biochemical kinetic colorimetric method. The average age of participants was 34.6 years (± 8.5). The most prevalent form of contact is via the application of the product (88.9%). The mean duration of exposure was 10.7 years. Still, 70.4% reported using personal protective equipment (PPE), more frequent use of mask (55.5%). The average values for cholinesterase was 3244.45 U/l (± 345.8) levels below those of the reference. The results obtained in this study are essential to use biological monitoring means of rural workers in purpose of prevention and health promotion.

Serum ß-glucuronidase activity as a biomarker for acute cholinesterase inhibitor pesticide poisoning.

ß-glucuronidase (BG) activity is a promising biomarker for diagnosis and prognosis after exposure to organophosphorous (OP) pesticides. The aim of this study was to evaluate the changes in serum BG activity in patients with acute OP poisoning and to determine whether these changes correlate with the severity of poisoning. Thirty patients with anticholinesterase pesticide poisoning were included, besides 10 healthy volunteers as a control group. Serum activities of butyrylcholinesterase (BuChE) and BG were measured for each subject on admission, then after 12 and 24 h. Serum levels of BuChE and BG in poisoned patients were significantly different from the control subjects; these differences persisted in repeated measurements. Moreover, the serum levels showed significant differences within each group of the three time points. A significant negative correlation was found between the serum activities of BuChE and BG in all groups at the three time points. In conclusion, serum BG activity seems a reliable marker for OP poisoning even when measured at 24 h after poisoning.

Verification of organophosphorus pesticide poisoning: Detection of phosphorylated tyrosines and a cysteine-proline disulfide-adduct from human serum albumin after intoxication with dimethoate/omethoate.

A method is described allowing forensic analysis of plasma samples to prove human poisoning with the organophosphorus pesticides omethoate (OM) and dimethoate (DIM). Upon incubation of human serum albumin (HSA) with both pesticides tyrosine residues were phosphorylated. In addition, a novel disulfide-adduct between the identical thiol-containing leaving group of OM and DIM (2-mercapto-N-methylacetamide, MNMA) and the only free cysteine residue in HSA (Cys34) was formed. Following pronase-catalyzed proteolysis either O,O-dimethyl phosphotyrosine (Tyr-dmp) or O,O-dimethyl thiophosphotyrosine (Tyr-dmsp) as well as the cysteine-proline dipeptide disulfide-adduct (MNMA-CysPro) were produced. All biomarkers were simultaneously detected using modern microbore liquid chromatography-electrospray ionization high-resolution tandem-mass spectrometry (µLC-ESI MS/HR MS). Corresponding limits of identification (LOI) for tyrosine-adducts (LOIOM: 30 µM, LOIDIM: 120 µM) and disulfide-adducts (LOIOM: 1.2 µM, LOIDIM: 30 µM) demonstrated that MNMA-CysPro allowed a considerably more sensitive detection. Finally, this novel method was applied to a plasma sample of an 87-year-old man, who had unintentionally ingested the pesticide Roxion® containing DIM as active ingredient. Unambiguous proof of poisoning demonstrated suitability of the novel biomarkers for sensitive verification analysis.

Prolonged paralysis in a child with organophosphate pesticide poisoning.

A 17-month-old boy presented to a local community health centre in Cape Town, South Africa, with severe organophosphate pesticide poisoning (OPP), necessitating the use of intravenous atropine to control cholinergic symptoms, as well as emergency intubation for ongoing respiratory distress. He required prolonged ventilatory support in the intensive care unit at his referral hospital and had subsequent delayed neurological recovery, spending 8 days in hospital.We present this case to emphasise the importance of adequate atropinisation in the management of severe OPP and to highlight the dangers of inappropriate use of suxamethonium for intubation in patients with OPP.

Novel cysteine- and albumin-adduct biomarkers to prove human poisoning with the pesticide oxydemeton-S-methyl.

We herein report on the forensic analysis of plasma samples to prove human poisoning with oxydemeton-S-methyl (ODM), S-(2-(ethylsulfinyl)ethyl)-O,O-dimethyl phosphorothioate. This organophosphorus pesticide is the active ingredient of Metasystox®, that was swallowed by a 77-year-old woman to commit suicide. ODM belongs to the class of dimethyl phosphoryl (DMP) pesticides, contains a 2-(ethylsulfinyl)ethanethiol (ESOET) leaving group and undergoes adduct formation with endogenous molecules as elaborated herein with human serum exposed to pesticides in vitro. A novel bioanalytical micro liquid-chromatography-electrospray ionization tandem high-resolution mass spectrometry method (µLC-ESI MS/HR MS) was developed to target multiple biomarkers of exposure. Following pronase-catalyzed proteolysis of patient plasma and subsequent ultrafiltration, the filtrate was analyzed. Diverse reaction products of ODM as well as of its oxidized biotransformation product demeton-S-methyl sulfone (DSMS), that possesses a 2-(ethylsulfonyl)ethanethiol (ESO2ET) leaving group, were simultaneously detected. Phosphorylated tyrosine residues (Tyr-DMP) derived from human serum albumin (HSA) as well as novel dipeptide-adducts containing the Cys34 residue of HSA coupled to ESOET and ESO2ET via a disulfide bond (ESOET-CysPro and ESO2ET-CysPro) were found. In addition, a related disulfide-product was detected comprising the single amino acid cysteine and ESOET (ESOET-Cys). Whereas Tyr-DMP only proved the intake of any DMP pesticide in general, its simultaneous detection with ESOET-CysPro, ESO2ET-CysPro and ESOET-Cys allowed unambiguous identification of the ingested pesticide. Therefore, the novel biomarkers and the method developed expand the possibilities of forensic investigations of ODM poisoning.

A toolbox for microbore liquid chromatography tandem-high-resolution mass spectrometry analysis of albumin-adducts as novel biomarkers of organophosphorus pesticide poisoning.

Exposure to toxic organophosphorus pesticides (OPP) represents a serious problem in the public healthcare sector and might be forced in terroristic attacks. Therefore, reliable verification procedures for OPP-intoxications are required for forensic, toxicological and clinical reasons. We developed and optimized a toolbox of methods to detect adducts of human serum albumin (HSA) with OPP considered as long-term biomarkers. Human serum was incubated with diethyl-oxono and diethyl-thiono pesticides for adduct formation used as reference. Afterwards serum was subjected to proteolysis using three proteases separately thus yielding phosphorylated tyrosine residues (Y*) detected as single amino acid (pronase), as hexadecapeptide LVRY*411TKKVPQVSTPTL (pepsin) and as the tripeptide Y*411TK (trypsin), respectively. Adducts were analyzed via microbore liquid chromatography coupled to electrospray ionization (µLC-ESI) and tandem-high-resolution mass spectrometry (MS/HR MS). Using paraoxon-ethyl as model OPP for adduct formation, methods were optimized with respect to MS/HR MS-parameters, protease concentrations and incubation time for proteolysis. HSA-adducts were found to be stable in serum in vitro at +37 °C and -30 °C for at least 27 days and resulting biomarkers were stable in the autosampler at 15 °C for at least 24 h. Limits of identification of adducts varied between 0.25 µM and 4.0 µM with respect to the corresponding pesticide concentrations in serum. Applicability of the methods was proven by successful detection of the adducts in samples of OPP-poisoned patients thus demonstrating the methods as a reliable toolbox for forensic and toxicological analysis.

Diagnostic value of complete blood count in paraquat and organophosphorus poisoning patients.

Complete blood count (CBC) is one of the most extensively used tests in clinical practice. In order to determine the diagnostic value of the CBC in paraquat (PQ) and organophosphorus (OPPs) poisoning, the CBC indices of PQ- and OPPs-poisoned patients were investigated in this study. A total of 96 PQ poisoning patients, 90 OPPs poisoning patients, and 188 healthy subjects were included in this study. The PQ- and OPPs-poisoned patients were divided into different groups according to their clinical symptoms. All CBC indices were analyzed by Fisher discriminant, partial least-squares discriminant analysis (PLS-DA), variance analysis, and receiver operating characteristic (ROC). The discriminant results showed that 87.7% of original grouped cases correctly classified between PQ-poisoned patients, OPPs-poisoned patients, and healthy subjects. The PLS-DA results showed that the important variable order was different in PQ- and OPPs-poisoned patients. Both white blood cell (WBC) and neutrophil (NE) counts were the most important indexes in PQ- and OPPs-poisoned patients. In OPPs poisoning patients, WBC and NE showed statistical differences between the severe poisoning group and the moderate poisoning group. Their areas under the ROC curve (AUC) were 0.673 (WBC) and 0.669 (NE), which were higher than cholinesterase (CHE; AUC 0.326). In conclusion, the CBC indices had a diagnostic value in PQ and OPPs poisoning; WBC and NE were the first responses and had clinical significance in PQ and OPPs poisoning; moreover, they are better than CHE in diagnosing OPPs poisoning.

Evaluation of chromosomal DNA damage, cytotoxicity, cytostasis, oxidative DNA damage and their relationship with endocrine hormones in patients with acute organophosphate poisoning.

Pesticides are commonly used compounds in agriculture. Especially, organophosphates (OPs) are among the extensively used pesticides. Therefore, OPs poisoning is common, especially in underdeveloped and developing countries. Primary aim of this study was to research the effects of acute OPs poisoning on genome instability in the individuals' lymphocytes with acute OPs poisoning both by using the cytokinesis-block micronucleus cytome (CBMN-cyt) assay to examine chromosome/genome damage, cell proliferation index and cell death rate and by using the plasma 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels to determine oxidative DNA damage. Secondary aim of this study was also to assess whether a relation exists between endocrine hormones and the genome damage in acute OPs poisoning. In the study, blood samples were analysed of 13 patients before and after treatment admitted to the Department of Intensive Care Unit with acute OPs poisoning and of 13 healthy subjects of similar age and sex. The present study demonstrates that genome damage (micronucleus; MN and nucleoplasmic bridges; NPBs frequencies), apoptotic and necrotic cell frequencies increased in lymphocytes of patients with acute OPs poisoning before treatment and decreased after treatment. The present study also show that CBMN cyt assay parameters and 8-OHdG levels could be affected by some endocrine hormones such as E2, fT3, fT4, GH, IGF-1, FSH, LH, TSH, PRL, but not be related to ACTH and tT levels in acute OPs poisoning. In conclusion, it is believed that this is the first study to evaluate the chromosomal/oxidative DNA damage, cell proliferation, cell death and their associations with endocrine hormones in acute OPs poisoning. These preliminary findings need to be supported by further studies with larger sample sizes.