A Rapid Mercury Droplet Electrode Polarography Method for Determination of Blood Lead Level in Lead Poisoned People.

BACKGROUND: Lead is a hazardous heavy metal, which causes many problems in the human body. Unfortunately, recent reports showed that smugglers and opium sellers add lead to drugs during the production procedure in order to increase its weight and cost. PURPOSE: The aim of this study was development of a rapid and accurate method for measurement of blood lead levels (BLL) in the oral and inhaled opiate abuser people. METHODS: BLL in samples obtained from the oral and inhaled opium addicted patients referring to Sina Hospital in Tabriz, Iran, during 2017 was compared with healthy control group (N=15). The wet digestion method was used to prepare whole blood and Mercury Droplet Electrode Polarography (MDEP) method was utilized for measurement of the lead content of digested samples. RESULTS: Results showed that there were significant differences between the BLL of samples obtained from oral (17.12±74.61 µg/dL, p<0.0003) and inhaled (19.33±2.257 µg/dL, p<0.0001) opium addicted groups in comparison with healthy control group (4.669±0.3367 µg/dL). CONCLUSION: Based on the results of this study it was observed that BLL in opium addicted people needs to be measured as soon as possible. Furthermore, screening of blood lead concentrations in opium-addicted people with a rapid and accurate MDEP method is very necessary and important.

High-Resolution FluoRespirometry and OXPHOS Protocols for Human Cells, Permeabilized Fibers from Small Biopsies of Muscle, and Isolated Mitochondria.

Protocols for High-Resolution FluoRespirometry of intact cells, permeabilized cells, permeabilized muscle fibers, isolated mitochondria, and tissue homogenates offer sensitive diagnostic tests of integrated mitochondrial function using standard cell culture techniques, small needle biopsies of muscle, and mitochondrial preparation methods. Multiple substrate-uncoupler-inhibitor titration (SUIT) protocols for analysis of oxidative phosphorylation (OXPHOS) improve our understanding of mitochondrial respiratory control and the pathophysiology of mitochondrial diseases. Respiratory states are defined in functional terms to account for the network of metabolic interactions in complex SUIT protocols with stepwise modulation of coupling control and electron transfer pathway states. A regulated degree of intrinsic uncoupling is a hallmark of oxidative phosphorylation, whereas pathological and toxicological dyscoupling is evaluated as a mitochondrial defect. The noncoupled state of maximum respiration is experimentally induced by titration of established uncouplers (CCCP, FCCP, DNP) to collapse the protonmotive force across the mitochondrial inner membrane and measure the electron transfer (ET) capacity (open-circuit operation of respiration). Intrinsic uncoupling and dyscoupling are evaluated as the flux control ratio between non-phosphorylating LEAK respiration (electron flow coupled to proton pumping to compensate for proton leaks) and ET capacity. If OXPHOS capacity (maximally ADP-stimulated O2 flux) is less than ET capacity, the phosphorylation pathway contributes to flux control. Physiological substrate combinations supporting the NADH and succinate pathway are required to reconstitute tricarboxylic acid cycle function. This supports maximum ET and OXPHOS capacities, due to the additive effect of multiple electron supply pathways converging at the Q-junction. ET pathways with electron entry separately through NADH (pyruvate and malate or glutamate and malate) or succinate (succinate and rotenone) restrict ET capacity and artificially enhance flux control upstream of the Q-cycle, providing diagnostic information on specific ET-pathway branches. O2 concentration is maintained above air saturation in protocols with permeabilized muscle fibers to avoid experimental O2 limitation of respiration. Standardized two-point calibration of the polarographic oxygen sensor (static sensor calibration), calibration of the sensor response time (dynamic sensor calibration), and evaluation of instrumental background O2 flux (systemic flux compensation) provide the unique experimental basis for high accuracy of quantitative results and quality control in High-Resolution FluoRespirometry.

Monitoring in microvascular tissue transfer by measurement of oxygen partial pressure: four years experience with 125 microsurgical transplants.

In a prospective study, the characteristics and benefit of an invasive measurement of oxygen partial pressure (pO(2)) with the aid of a polarographic sensor were investigated in 125 microsurgical reconstructions of the head and neck area over a period of 45 months. Measurements were performed over 96 h in eight different types of microsurgically revascularized flaps for extra- and intraoral reconstructions and were evaluated separately for each flap type. Of 125 reconstructions the system indicated malperfusion in 18 cases. Salvage surgery was performed in 17 cases due to venous thrombosis (6 cases), arterial thrombosis (3 cases), a combination of arterial and venous thrombosis (2 cases), rheological problems (3 cases), venous insufficiency by hematoma (2 cases) and kinking of vessels (1 case). In 10 cases salvage surgery was successful, 7 flaps were lost despite salvage surgery. In all these cases, the polarographic probe indicated the necessity of salvage surgery correctly. After 96 h no malperfusion was seen. Postoperatively, a common and characteristic development of the oxygen partial pressure in different types of flaps was seen. Initially, a clear increase of pO(2) could be measured. During 96 h, a slow decrease of pO(2) was observed. In conclusion polarographic measurement of pO(2) can be an excellent apparative supplement for the postoperative clinical control of microsurgically revascularized transplants. In buried flaps, this technique represents the only reliable method for transplant monitoring.

Brain tissue oxygen pressure monitoring in awake patients during functional neurosurgery: the assessment of normal values.

Local brain tissue oxygen (ptiO2) monitoring is frequently applied in patients at risk for cerebral ischemia. To identify ischemic thresholds, the normal range of local brain tissue oxygen pressure (ptiO2) values needs to be established. Ideally, such normal values are determined in healthy and awake subjects, so as to eliminate the possible influences of anesthetics on cerebral physiology or ptiO2. Thus far, however, such measurements have not been conducted, and to fill this void, we determined the ptiO2 values in normal white matter of awake patients undergoing functional stereotactic brain surgery. In 25 otherwise healthy patients, who underwent functional neurosurgery for treatment of a refractory movement disorder under local anesthesia, the ptiO2 of white matter was recorded continuously using a polarographic Clark type electrode monitoring system. Preoperative screening ruled out cognitive dysfunction or structural cerebral lesions. Reliable intraoperative ptiO2 values were obtained in 22 patients. After an adaptation period of 118+/-35 min (range, 47-171 min), we found an average normal ptiO2 of 22.6+/-7.2 mm Hg in the frontal white matter. In 11 patients, ptiO2 measurements were continued postoperatively for 24 h. During this period, a similar normal ptiO2 value of 23.1+/-6.6 mm Hg was found. No iatrogenic complications occurred. In conclusion, the normal ptiO2 of cerebral white matter is most likely lower than previously assumed. Further, the long adaptation time renders this widely applied monitoring instrument unreliable in detecting ischemia early after insertion and limits its usefulness for intraoperative monitoring.

Prediction of avascular necrosis of the femoral head by measuring intramedullary oxygen tension after femoral neck fracture.

OBJECTIVE: To measure the intramedullary oxygen tension of the proximal femur after a femoral neck fracture and to evaluate the usefulness of that monitoring for prediction of subsequent avascular necrosis. DESIGN: Prospective case series with comparison group. SETTING: University hospital. INTERVENTION: Measurement of intramedullary oxygen tension of the femoral head and neck during internal fixation using the Hansson hook-pin system. MAIN OUTCOME MEASUREMENTS: Intramedullary oxygen tension was measured directly during surgery in 17 patients with 18 femoral neck fractures treated by internal fixation between October 2000 and February 2002. The intramedullary oxygen tension was measured by using polarographic oxygen electrodes and an oxygen monitor at four points: (A) 1 cm distal from the joint surface; (B) 1 cm proximal from the fracture site; (C) 1 cm distal from the fracture site; and (D) 1 cm proximal from the lateral wall. The presence or absence of avascular necrosis was evaluated by magnetic resonance imaging (MRI) at 2, 6, and 12 months after surgery. RESULTS: MRI evaluation showed 11 fractures healed without complications, and 7 fractures developed avascular necrosis. We found significant differences in the distribution of intramedullary oxygen tension of the femoral head between points A (1 cm from the joint surface) and B (1 cm proximal from the fracture site) in those patients who developed avascular necrosis (P = 0.039); that is the oxygen tension was lower at point A than at point B. In contrast, in those patients who did not develop avascular necrosis there was no significant differences between point A and B were found (P = 0.059). The sensitivity and specificity for prediction of avascular necrosis were 1.0 and 0.82 (Fischer exact probability test, P = 0.002), respectively, when the cut-off level of oxygen tension differences between points A and B was set at 3.1 mm Hg. CONCLUSION: We believe that this method of measuring intramedullary oxygen tension is simpler and less invasive than other currently used methods and has the possibility for intraoperatively identifying a risk group that can develop a late segmental collapse of the femoral head secondary to avascular necrosis.

Reactions of sulfenic acid with 2-mercaptoethanol: a mechanism for the inhibition of gastric (H+-K+)-adenosine triphosphate by omeprazole.

The reactions of omeprazole, a potent proton pump inhibitor (PPI) were investigated in the presence of 2-mercapotoethanol. Reactions were monitored in solutions buffered to pH values ranging 2.0-8.0 using differential pulse polarography (DPP) at the static mercury drop electrode (SMDE). The fast, sensitive and selective electrochemical technique facilitated successive recordings of voltammograms (peak current (nA) vs. peak potential (volts vs. Ag/AgCl)) for all analytes in situe, including the 2-mercaptoethanol. In acidic solutions and in the presence of 2-mercaptoethanol, omeprazole undergoes degradation into three compounds, the first is a cyclic sulfenamide (D+), previously believed to be the active inhibitor of the H+, K+-ATPase, the second is the omeprazole dimer, and the third is the disulfide believed to be the product of reaction between 2-mercaptoethanol and D+. The cyclic sulfenamide (D+) solution was found to be stable in solutions containing 2-mercaptoethanol having pH values: 2.0, 4.0, and 6.0. This finding proved conclusively that the cyclic sulfenamide is not reactive toward the 2-mercaptoethanol. In contrast to previous reports, the conversion of the sulfenic acid intermediate into D+ was found to be irreversible. Due to this irreversibility, D+ and sulfenic acid were not rapidly interconvertable. The present work suggests that the active inhibitor is the sulfenic acid.

Myocardial oxygen tension during surgical revascularization. A clinical comparison between blood cardioplegia and crystalloid cardioplegia.

OBJECTIVE: The aim of this study was to assess the effect of cardioplegic solutions on myocardial oxygenation during surgical revascularization. METHODS: In 30 patients, randomized to receive crystalloid (CC) or blood (BC) cardioplegia, myocardial oxygen tension was measured continuously by polarography. RESULTS: The two groups were comparable in terms of patients' age, sex, pre-operative ejection fraction, coronary disease, perfusion time, and aorta cross-clamping time. However, the BC group required 22% more of cardioplegic solution to stop electrical activity of the heart. Throughout the pre- and post-cardiac arrest periods, oxygen tension between the two groups was similar. At the end of the observation (4th day), myocardial oxygenation increased over 200% in relation to the values before revascularization. During the first infusion of cardioplegia, oxygen tension in the CC group was lower compared to the BC group (0.1 mmHg vs 1.3 mmHg; P<0.05) being the only significant difference between the two groups during cardiac arrest. Throughout the cardiac arrest, myocardial oxygen tension was close to zero regardless of the type of cardioplegia used. Post-operatively, addition of oxygen to the respiratory air increased myocardial oxygenation by over 17% resulting in a positive correlation (r=0.94; P<0.05) between myocardial oxygen tension and peripheral saturation. CONCLUSIONS: In conclusion, the differences in myocardial oxygen tension between the CC and BC groups are trivial. Thus, any potential beneficial effect of blood cardioplegia compared to crystalloid cardioplegia must be due to other circumstances than its oxygen carrying capacity. An important observation is a significant increase in myocardial oxygenation during oxygen supplement to the respiratory air.

Measurement of oxygen partial pressure in the mandibular bone using a polarographic fine needle probe.

In this study, the oxygen partial pressure in the cancellous bone substance of the mandible was measured for the first time with a polarographic fine needle probe. This has so far only been established in soft tissue. The aim was to prove and to test the feasibility in principle of this method of measurement in order to ascertain the normal values for the O2 partial pressure in healthy bone. These values were afterwards compared with the results of measurements in areas of different pathological bone conditions in order to assess the clinical suitability of the method for "mapping" during mandibular resection. Measurements of oxygen partial pressure were made in a total of 42 patients (16 women, 26 men). Of these, 12 patients with clinically normal bones served as a control group. Seventeen patients had osteoradionecrosis following radiation treatment, and 13 patients presented with chronic osteomyelitis of the mandible. All measurements were carried out with a polarographic fine needle probe applied to the cancellous bone substance. The statistical analysis included a comparison of the mean values of the oxygen partial pressures measured. No statistical correlation between oxygen partial pressure and pH and hemoglobin values could be detected. The average oxygen partial pressure in the healthy mandibular bone was 71.4 mmHg. In non-healthy bone, the value fell to an average of 30.6 mmHg (osteoradionecrosis 32.3 mmHg, chronic osteomyelitis of the mandibular bone 28.4 mmHg). Statistically, the differences in the group values ascertained were highly significant (P < 0.005). The data found show that this method can be successfully used to detect poorly perfused bone. The values are reproducible and reflect the clinical situation. In the long term, the method appears to be a suitable diagnostic tool for assessing the oxygen supply in bone in studying various clinical problems related to bone surgery.

Intramyocardial oxygen monitoring in coronary artery bypass surgery.

OBJECTIVE: In coronary artery bypass surgery various parameters have been used to monitor patients clinical status. Direct monitoring of myocardial oxygenation can be performed by measuring intramyocardial partial oxygen tension pressure (p ti O2). This study was performed to determine the perioperative time course of this parameter in correlation to standard monitoring parameters. METHODS: Twenty-three patients underwent standard coronary artery bypass grafting (CABG). A special polarographic microprobes was inserted into the myocardium in the distribution zone of the left anterior descending artery which was one of the target vessels of myocardial revascularization. Intramyocardial p ti O2 was monitored intra- and up to 12 hours postoperatively. Values were correlated to hemodynamic, oxygenation and procedure associated parameters. RESULTS: Myocardial oxygenation during CABG is characterized by a significant decrease of p ti O2 during cross-clamping and a significant increase after removal of the cross-clamp. The postoperative time course of p ti O2 shows a steady increase of p ti O2 in the first 12 postoperative hours investigated. Preoperative ejection fraction as well as cardio-pulmonary bypass time does not seem to have an influence on the postoperative p ti O2 in these patients. Various standard monitoring parameters show complex influence on intramyocardial p ti O2- CONCLUSIONS: Determination of intramyocardial partial oxygen pressure in patients undergoing bypass surgery shows characteristic changes. Changes in p ti O2 as a direct online parameter of myocardial oxygenation occur immediately after procedures that influence myocardial perfusion and therefore, may help to detect potential complications earlier than standard monitoring parameters in cardiac surgery.

Near infrared spectroscopy and imaging to probe differences in water content in normal and cancer human prostate tissues.

The content of water in cancerous and normal human prostate in vitro tissues was shown to be different using near infrared (NIR) spectroscopy. The water absorption peaks at 1444 nm and 1944 nm are observed in both types of prostate tissues. The measurements show that less water is contained in cancerous tissues than in normal tissues. The OH stretching vibrational overtone mode at 1444 nm and other water overtone modes provide key spectroscopic fingerprints to detect cancer in prostate tissue. Transmission and backscattered spectral imaging were measured in cancer and normal prostate tissues. The degree of polarization for 700 nm, 800 nm, 1200 nm, and 1450 nm is larger for normal than for cancer tissues. The knowledge about water content offers a potential as a diagnostic tool to better determine and image cancer in prostate and in other tissues types such as breast and cervix using the absorption from vibrational overtones of H(2)O molecules in the NIR.

Estimating hypoxic status in human tumors: a simulation using Eppendorf oxygen probe data in cervical cancer patients.

PURPOSE: To define the minimal number of pO(2) measurements, with 90% sensitivity and 90% specificity, needed to categorize cervical tumors as either hypoxic or oxic. METHODS AND MATERIALS: Using Eppendorf oxygen probe data from our ongoing prospective trial, we simulated the measurement of tumor oxygenation with a smaller number of data points in 135 patients with cervical cancer. The hypoxic proportion, defined as the percentage of pO(2) values <5 mm Hg (HP5), was calculated for each tumor. Hypoxic tumors were defined as those with a median HP5 >50%, and tumors with normal oxygen levels as those with a median HP5 < or =50%. A small number of pO(2) measurements were randomly selected from the Eppendorf measurements in each tumor, or per Eppendorf track, and used to define the tumor as hypoxic or oxic. The sensitivity and specificity were calculated, considering the classification as given by the complete set of Eppendorf measurements as the reference standard. RESULTS: The probability of falsely classifying the tumor decreased as the selected number of pO(2) measurements per tumor increased, and at 16 measurements was approximately 10%. Adding additional measurements per tumor beyond 24 improved the ability to classify the tumor accurately only slightly. The probability of falsely classifying the tumor decreased as the pO(2) measurements per track increased. At five measurements per track, the probability of falsely classifying the tumor was approximately 9%. CONCLUSION: Approximately 20 measurements per tumor, or five measurements per track, using the Eppendorf pO(2) histograph, are sufficient to categorize cervical tumors as hypoxic or oxic. The results of this study will serve as a guide for research clinicians in the use of this and other systems in the assessment of tumor oxygenation in humans.

Determination of cadmium, lead, copper and zinc in Yemeni khat by anodic stripping voltammetry.

Trace element concentrations in khat were investigated as they can disturb trace element levels in the body. Cadmium (Cd), lead (Pb), copper (Cu) and zinc (Zn) levels in khat and 6 leafy vegetables commonly consumed in the Republic of Yemen were determined by differential pulse anodic stripping voltammetry after wet digestion of the organic matter. Khat had significantly higher concentrations of Cu and Zn than did the leafy vegetables, but similar amounts of Cd and Pb. The average daily intake of khat consumers of Cd, Pb, Cu and Zn from khat only was estimated to be 2.0-10.2 microg/day, 23.6-118.0 microg/day, 530-2654 microg/day and 662-3311 microg/day respectively. Although high, these values were within Food and Agriculture Organization/World Health Organization tolerance limits.

Comparison of tumor and normal tissue oxygen tension measurements using OxyLite or microelectrodes in rodents.

In this study we compare oxygen tension (PO2) histograms measured with O2 microelectrodes and a new optical PO2 measurement device, the OxyLite, in normal tissues (mouse spleen and thymus) and in tumors (R3230Ac in rats) (n = 5-6). The transient response to glucose infusion or 100% O2 breathing (hyperoxia) was also measured in tumors. PO2 histograms of spleen and thymus with the two devices were not different. The OxyLite tumor PO2 histogram, however, was left-shifted compared with the microelectrode (median PO2 1.0 vs. 4.0 mmHg, P = 0.016). Both probes responded to acute hyperglycemia with a mean increase of 3-6 mmHg, but the microelectrode change was not significant. The OxyLite consistently recorded large PO2 increases (approximately 28 mmHg) with hyperoxia, whereas the microelectrode response was variable. The OxyLite averages PO2 over an area that contains interstitial and vascular components, whereas the microelectrode measures a more local PO2. This study demonstrates the importance of considering the features of the measurement device when studying tissues with heterogeneous PO2 distributions (e.g., tumors).

Head and neck cancer: the importance of oxygen.

OBJECTIVES: To use recently introduced polarographic technology to characterize the distribution of oxygenation in solid tumors, explore the differences between severe hypoxia and true necrosis, and evaluate the ability to predict treatment outcomes based on tumor oxygenation. STUDY DESIGN: Prospective, nonrandomized trial of patients with advanced head and neck cancer, conducted at an academic institution. METHODS: A total of 63 patients underwent polarographic oxygen measurements of their tumors. Experiment 1 was designed to determine whether a gradient of oxygenation exists within tumors by examining several series of measurements in each tumor. Experiment 2 was an analysis of the difference in data variance incurred when comparing oxygen measurements using oxygen electrodes of two different sizes. Experiment 3 compared the proportion of tumor necrosis to the proportion of very low (< or =2.5 mm Hg) polarographic oxygen measurements. Experiment 4 was designed to explore the correlation between oxygenation and treatment outcomes after nonsurgical management. RESULTS: No gradient of oxygenation was found within cervical lymph node metastases from head and neck squamous cell carcinomas (P > .9). Tumor measurements achieved with larger (17 microm) electrodes displayed smaller variances than those obtained with smaller (12 microm) electrodes, although this difference failed to reach statistical significance (P = .60). There was no correlation between tumor necrosis and the proportion of very low (< or =2.5 mm Hg) oxygen measurements. There was a nonsignificant trend toward poorer locoregional control and overall survival in hypoxic tumors. CONCLUSIONS: Hypoxia exists within cervical lymph node metastases from head and neck squamous carcinomas, but the hypoxic regions are distributed essentially randomly. As expected, measurements of oxygen achieved with larger electrodes results in lowered variance, but with no change in overall tumor mean oxygen levels. Polarographic oxygen measurements are independent of tumor necrosis. Finally, oxygenation as an independent variable is incapable of predicting prognosis, probably reflecting the multifactorial nature of the biological behavior of head and neck cancers.

Effects of experimental cochlear thrombosis on oxygenation and auditory function of the inner ear.

To elucidate the etiology and pathogenesis of sudden hearing loss, the effect of experimental cochlear thrombosis on oxygenation and the auditory function of the inner ear was investigated in anesthetized guinea pigs. Impairment of cochlear blood flow (CBF) was induced by ferromagnetic obstruction of cochlear blood vessels at lowered body temperature. Perilymphatic oxygen partial pressure (PO2) in the basal scala tympani (about 200 microm below the round window membrane) was measured polarographically using micro-coaxial needle electrodes. Auditory function was examined by recording cochlear microphonic (CM) frequency responses, compound action potentials (CAP) and auditory evoked brainstem responses (ABR). Findings demonstrated a considerable decrease in the mean perilymphatic PO2 of 40%, 2 h after the start of the experiment. Mean CM and N1 CAP amplitudes were reduced by about 25% each and ABR by 18%. No significant changes were observed in the latencies of either CAP or ABR. Mean basal CBF was found to decrease by 35%, as measured by laser Doppler flowmetry in a parallel study. The present findings demonstrate that vascular impairment in the inner ear results in a considerable drop in intracochlear oxygenation, causing a significant loss in the auditory response.

Adverse effects of hyperoxemia during cardiopulmonary bypass.

OBJECTIVE: Aiming at elucidating the effects on capillary blood flow and tissue oxygenation of hyperoxemia during cardiopulmonary bypass, we studied skeletal muscle surface oxygen tensions in 10 patients undergoing elective cardiac operations. METHODS: In a prospective investigation each patient was exposed to normoxemia (arterial oxygen tension 75 to 115 mm Hg) and hyperoxemia (arterial oxygen tension > 185 mm Hg, inspired oxygen fraction = 1.00) during normal anesthetized conditions before and after cardiopulmonary bypass, as well as during normothermic and hypothermic continuous-flow bypass. In each state hemodynamic variables and arterial and mixed venous blood gas and acid base values were measured. From these data oxygen transport variables were calculated. Tissue oxygenation was studied with the use of a multiple-point polarographic oxygen microelectrode, known to provide measures of oxygen tensions at the capillary level. The oxygen distribution profile of such a sample is also indicative of capillary blood flow distribution changes. RESULTS: In all patients and at each occasion of the investigation markedly low mean surface oxygen tensions in skeletal muscle were registered. When hyperoxemia was instituted, a significant decrease in these surface oxygen tensions together with an increase in distribution heterogeneity was seen during all stages. Contrary to prebypass, postbypass, and hypothermic bypass, where vascular resistance, oxygen delivery, and oxygen consumption remained similar during hyperoxemia and normoxemia, a significant (p < 0.05) increase in vascular resistance together with a decline in oxygen consumption was seen during hyperoxemic normothermic (35 degrees to 36 degrees C) cardiopulmonary bypass. CONCLUSION: These findings show that the microcirculatory response to hyperoxemia, seen under other circumstances, persists during continuous-flow cardiopulmonary bypass, normothermic as well as hypothermic. If these adverse effects on tissue oxygenation by hyperoxemia can be further verified and shown to be valid for other organs than skeletal muscle, we would suggest that hyperoxemia should be avoided, especially during normothermic cardiopulmonary bypass.

The effect of the perflubron emulsion Oxygent on the calibration characteristics of polarographic oxygen electrodes.

The perfluorocarbon emulsion Oxygent improves normal tissue oxygenation. A clinical trial is planned to evaluate this effect in tumors. Polarographic oxygen electrodes were calibrated in solutions of saline +/- perflubron to assess whether Oxygent influenced probe behavior. Calibrations were linear under both conditions. There were no major differences in estimated oxygen tensions.