The effects of heavy metals on common carp white blood cells in vitro

The in vitro effects of cadmium, copper, lead and zinc, and various cadmium compounds (chloride, sulphate and nitrate) on common carp (Cyprinus carpio) lymphocyte viability and phagocyte activity, were evaluated. The percentage of dead lymphocytes was determined after Trypan blue staining, and phagocyte activity was measured by using the nitroblue tetrazolium (NBT) reduction test. Lead was the most toxic to lymphocytes--the maximum mortality exceeded 30%, and was significantly higher at 1 microM of lead, compared to the control. The maximum mortality caused by cadmium was below 10%, but was significantly elevated with 5 microM or more of cadmium. Zinc induced lymphocyte mortality from 10 microM, whilst no effect was observed with copper. The incubation of full blood with the three cadmium compounds (at 5mg/l of cadmium) showed that cadmium nitrate and cadmium sulphate were more toxic (over 35% and 25% mortality, respectively) than cadmium chloride (about 15% mortality). This was confirmed by the results of tests on isolated cells--1mg/l of cadmium as nitrate and sulphate increased lymphocyte mortality compared to the control and cadmium chloride. Phagocytic activity was less sensitive to heavy metals than was lymphocyte viability. It was significantly reduced following exposure to 50 microM and 100 microM cadmium, and 100 microM zinc, but no effects were observed with either lead or copper.     

Membrane perturbations induced by the interactions of zinc ions with band 3 in human erythrocytes

Of group 12 metals, zinc is an essential element to maintain our life, but other metals such as cadmium and mercury are toxic in cellular activities. Interactions of these metals with biomembranes are important to understand their effects on our living cells. Here, we describe the membrane perturbations induced by these metals in human erythrocytes. Of these metals, Zn²⁺ ions only induced the erythrocyte agglutination. Histidine residues in extracellular domains of band 3 participated in Zn²⁺-induced agglutination. Interestingly, it was found that band 3-cytoskeleton interactions play an important role in Zn²⁺-induced agglutination. In contrast with Hg²⁺ and Cd²⁺ ions, Zn²⁺ ions greatly suppressed pressure-induced hemolysis by cell agglutination. Such a suppression was removed upon dissociation of agglutinated erythrocytes by washing, indicating the reversible interactions of Zn²⁺ ions with erythrocyte membranes. Excimer fluorescence of pyrene indicated that spectrin is denatured by a pressure of 200 MPa irrespective of hemolysis suppression. Taken together, these results suggest that the agglutination of erythrocytes due to the interactions of Zn²⁺ ions with band 3 is stable under pressure, but spectrin, cytoskeletal protein, is denatured by pressure     

Determination of Human Exposure to Lead and Cadmium: A WHO/UNEP Pilot Study

Abstract

In Stockholm, methods for measuring exposure to lead and cadmium from air, food and beverages were studied in 1988 in a group of 15 non-smoking women, as part of the WHO/UNEP HEAL programme. Airborne particles in the breathing zone air (24-hour samples), duplicate diets (24-hour samples), and faeces (all the stools produced) were collected during 7 consecutive days. Blood was sampled before and immediately after the study period. The results confirmed the need for personal monitoring in the assessment of human exposure to lead and cadmium via air and food. There is need for suitable equipment for 24-hour personal air monitoring. On average, dietary lead (26 μg day^?1, SD 7.9) contributed more than 80% of the total lead uptake, while dietary cadmium (8.5 μg day^?1 SD 2.1) contributed about 99% of the total cadmium uptake. Occasionally consumed foodstuffs with high levels of lead or cadmium seemed to be responsible for a large part of the total weekly intake of lead and cadmium. Fecal lead and cadmium were found to be useful indicators of the total amounts of these metals ingested. Due to the large day-to-day variation observed in the dietary intake of lead and cadmium, the sampling period for duplicate diets and faeces should be at least 5–6 days.     

Dietary Exposure to Lead and Cadmium in Yugoslavia

Abstract

Lead exposure still represents a matter of health concern especially in Yugoslavia. To assess the exposure of normal urban population to lead and cadmium through food, a preliminary monitoring was performed on a small group of urban population. Lead, cadmium and some essential elements (calcium, zinc, iron, copper and manganese) were analysed in collected duplicate diet samples and compared to similar population in Sweden. We found that dietary exposure to lead and cadmium is similar to other countries although Yugoslav urban population is exposed to much higher concentrations of lead in air than in cities of developed countries, due to high lead in gasoline. However, daily intake of some essential elements was significantly lower.

Also populations living around lead smelters in various parts of Yugoslavia are still exposed to elevated environmental lead and cadmium levels. To assess the exposure of the population living in this area, a cumulative long-term exposure to lead was determined by measuring lead in deciduous teeth. Concentrations of lead and cadmium in vegetables, soil and meals from the same region were also analysed. Values obtained for lead and cadmium in food products grown in exposed and control area were found to be related to respective concentrations of these elements in soil as well as to the distance from the smelter. Meals prepared in this region show the same trend, revealing very high intake particularly of lead.

The influence of nutritional factors, i.e. dietary calcium on lead metabolism, was also studied. Blood lead concentration was determined in two groups of peasant women living in two regions with different dietary calcium intake. Lower blood lead values were found in the higher dietary calcium intake region.     

Chemical fractionation of heavy metals in soils around oil installations,Assam

Abstract

The chemical fractionation of lead, cobalt, chromium, nickel, zinc, cadmium and copper in soils around Lakwa oil field, Assam, India was studied using a sequential extraction method. It is evident from the study that the residual fraction is the most important phase for the seven heavy metals under study. Among non-residual fractions metals are mostly associated with the Fe–Mn oxides fraction. The association of heavy metals with organic matter was observed in the following order: copper> cadmium> zinc> lead. The concentration of Pb in the carbonate fraction for both the seasons is higher compared with other metals, which may pose environmental problems due to its highly toxic nature. The comparatively low concentration of metals in the exchangeable fraction indicates low bioavailability. Correlations between physicochemical parameters and metal fractions of soil do not show consistent behaviour. The local mean values of metals when compared with the accepted values of normal abundance and geochemical background, indicates two to four fold increases in this area. However, the values are within the range of normal abundance. As well as from natural soil geochemical behaviour, anthropogenic influence might have a close bearing on the association of metals with the soil system in the studied area.     

Increased frequency of delayed type hypersensitivity to metals in patients with connective tissue disease

BackgroundConnective tissue disease (CTD) is a group of inflammatory disorders of unknown aetiology. Patients with CTD often report hypersensitivity to nickel. We examined the frequency of delayed type hypersensitivity (DTH) (Type IV allergy) to metals in patients with CTD.MethodsThirty-eight patients; 9 with systemic lupus erythematosus (SLE), 16 with rheumatoid arthritis (RA), and 13 with Sjögren's syndrome (SS) and a control group of 43 healthy age- and sex-matched subjects were included in the study. A detailed metal exposure history was collected by questionnaire. Metal hypersensitivity was evaluated using the optimised lymphocyte transformation test LTT-MELISA^® (Memory Lymphocyte Immuno Stimulation Assay).ResultsIn all subjects, the main source of metal exposure was dental metal restorations. The majority of patients (87%) had a positive lymphocyte reaction to at least one metal and 63% reacted to two or more metals tested. Within the control group, 43% of healthy subjects reacted to one metal and only 18% reacted to two or more metals. The increased metal reactivity in the patient group compared with the control group was statistically significant (P < 0.0001). The most frequent allergens were nickel, mercury, gold and palladium.ConclusionsPatients with SLE, RA and SS have an increased frequency of metal DTH. Metals such as nickel, mercury and gold are present in dental restorative materials, and many adults are therefore continually exposed to metal ions through corrosion of dental alloys. Metal-related DTH will cause inflammation. Since inflammation is a key process in CTDs, it is possible that metal-specific T cell reactivity is an etiological factor in their development. The role of metal-specific lymphocytes in autoimmunity remains an exciting challenge for future studies.     

Using tree leaves and barks collected from contaminated and uncontaminated areas as indicators of air metallic pollution

Abstract

The aim of this study was to show the usefulness of woody species in assessing air pollution by cadmium, copper, lead, and zinc. For this, washed leaves, unwashed leaves, and barks of a coniferous species (Cupressus sempervirens var. fastigiata) and a broadleaved one (Eucalyptus cladocalyx F. Muell) were analyzed for cadmium, copper, lead and zinc contents. A transect sampling approach was carried out during spring 2016. Fifty stations were selected along an intensive traffic road, and three in a control area. The results showed that the highest concentrations were recorded in barks for copper (21.86?µg/g, 23.33?µg/g) and lead (14.53?µg/g, 63.33?µg/g), and in unwashed leaves for cadmium (0.57?µg/g, 1.19?µg/g) and zinc (48.94?µg/g, 47.6?µg/g) for E. cladocalyx F. Muell, and C. sempervirens var. fastigiata, respectively. Lead and zinc are the most accumulated metals compared to cadmium and copper in all samples. The studied metal contents in urban area were significantly higher than that of the control one. This represents that traffic road has influenced the metals contamination of the surrounding area. Results of the bioconcentration factor (BCF) for both species indicate that the studied species could be used as biomonitors to identify ecological problems and to predict effect on wildlife habitats. The highest values of metal accumulation index (MAI) indicate the effectiveness of these trees for controlling the air metals in the polluted areas. Result shows too that the studied species could be used for phytoextraction of heavy metals from the polluted soils and/or air.     

Utilization of Mentha aquatica L. for removal of fecal pathogens and heavy metals from water of Bosna river,Bosnia and Herzegovina

Abstract

The aim of the present study was to investigate the potential of Mentha aquatica L. for phytoremediation of water contaminated with heavy metals and fecal pathogens from Bosna river. The water was treated with M. aquatica for 5, 10, and 15 days consecutively after which it was analyzed for the various physicochemical and microbiological parameters.

The initial concentration of cadmium (Cd) ranged from 3.644 to 6.108?µg/l, while lead (Pb) varied between 0.1 and 1.386?µg/l. After treatment, M. aquatica accumulated significant amounts of cadmium (Cd) and lead (Pb) with the highest removal rates of 96.49% for Cd and 45.72% for Pb. Values of several physicochemical parameters were decreased after 15 days treatment period.

All water samples were analyzed for enumeration of aerobic heterotrophic bacteria, total coliforms, and fecal coliforms by the membrane filtration. Removal efficiency was greater than 80% for microbiological parameters. The concentration of heavy metals was determined in different plant parts and subsequently, the translocation factor was determined. In M. aquatica plant parts, concentrations of Pb and Cd were increased after 15 days of treatment. Our results demonstrated that M. aquatica could be good candidates for the removal of fecal pathogens and heavy metals present in surface water.     

Protective effect of quercetin on some hematological parameters in rats exposed to cadmium

ABSTRACT

We investigated the effects of quercetin (Q) on some hematological parameters and determined the percentage of alpha-naphthyl acetate esterase (ANAE) positive lymphocytes in rats that had been exposed to cadmium (Cd). Thirty male Wistar albino rats were divided into four groups: control (C), quercetin (Q), cadmium (Cd) and Q + Cd (CdQ). Blood samples were taken to assess erythrocytes (RBC), leukocytes (WBC), hemoglobin levels (Hb), hematocrit values (Hct), platelets (PLT), alpha-naphthyl acetate esterase (ANAE) positive lymphocytes. RBC, Hb, Hct; the number of PLT significantly decreased in the Cd group. To the contrary, these parameters were increased significantly in the CdQ group compared to the Cd group. Although we found a significant increase in total WBC count and neutrophil percentage, the number of lymphocytes decreased in the Cd group compared to the other three groups. Also, the percentage of peripheral blood ANAE positive lymphocytes decreased significantly in the Cd group (p < 0.05). Q exhibits positive effects on some hematological characteristics and the percentage of ANAE positive lymphocyte in cases of acute CD toxicity.     

Metallothionein induction in human peripheral blood lymphocytes by heavy metals

Human peripheral blood lymphocytes have the capacity to produce metallothioneins (MTs) as a protective response to cadmium exposure. To define the range of metal species inducing lymphocyte MTs, cellular proteins synthesized after exposure to each of 11 heavy metals were analyzed by gel electrophoresis. Toxic metals such as cadmium, mercury and silver were found to induce thioneins (apoproteins of MTs) at relatively low concentrations (maximum at approximately 10 microM), whereas less toxic metals such as zinc, copper and nickel were inductive at relatively high concentrations (maximum at approximately 200 microM). Tin, lead, iron, cobalt, and manganese did not induce thioneins. The heavy metal specificity of MT induction in the lymphocyte resembles that in the liver, and the regulatory mechanism of MT production seems to be similar in both of these tissues. In the cells exposed to highly toxic metals such as cadmium and mercury, expression of cytotoxicity (represented by decline of cysteine uptake) was remarkable at the metal concentrations higher than those saturating thionein induction, supporting the protective role of MTs against heavy metals.     

Association of electronic cigarette use with lead,cadmium, barium,and antimony body burden: NHANES 2015-2016

BackgroundExposure of toxic metals from e-cigarette use is a cause for public health concern because youth, young adults, and non-smokers are the target population rapidly adopting e-cigarette use. The purpose of this research is to determine the association of the body burden of heavy metals with e-cigarette use using NHANES (U.S.) 2015–2016 data.MethodsBlood lead (N = 1899) and urinary cadmium, barium, and antimony (N = 1302) data were extracted from NHANES, 2015–2016; geometric means were calculated and bivariate and multivariable linear regression analyses were conducted. Participants were categorized as having neither e-cigarette nor cigarette use; smoking history (including dual use with e-cigarettes); and only e-cigarette (current or former).ResultsIn multivariable analyses adjusted for sex, race/ethnicity, age, and poverty levels, current or former e-cigarette use failed to reach a statistical significance in the association with metals. However, participants with a smoking history were more likely to have higher blood lead and urinary cadmium than participants who neither used e-cigarettes nor cigarettes.ConclusionBlood lead levels, and urinary cadmium, barium, and antimony levels were similar between participants who used e-cigarettes and participants who did not.     

Heavy metal distribution in zooplankton from Buenos Aires coastal waters (Argentina)

ABSTRACT

Total Zooplankton samples from Argentinian and Uruguayan Coastal Waters were analysed to determine their heavy metal contents. Zinc, copper, cadmium, iron, lead and manganese concentrations were measured using atomic absorption spectrophotometry and metal content distributions were geographically plotted. Except for cadmium, all the studied metals showed maximum values on the Uruguayan shores, and this was related not only to the location of the potential metal sources, but also to the hydrographical characteristics of the area. Finally, the suitability of zooplankton as an adequate biological indicator of heavy metal pollution in coastal environments is discussed.     

Freezing injury to erythrocytes. II. Morphological alterations of cell membranes.

Morphological alterations of human red blood cell membranes were examined with the cells containing different concentrations of glycerol being subjected to rapid rates of cooling, approximately 10⁴ and 10⁵ °C/min, and subsequent rewarming. Small membrane defects, similar to holes, were observed in specimens frozen with and without 10% glycerol. Various degrees of roughness were found on the surface of the cells at all freezing rates tested. The membrane alterations were reduced with increasing glycerol concentration, although roughness also appeared on the surface of the cells in 30% glycerol suspensions, frozen rapidly, and rewarmed to ?80 or ?60 °C. The cell membrane surface texture correlated with the growth of intra- and extracellular ice particles. There was also a positive correlation between these alterations and post-thaw hemolysis. It is concluded, therefore, that morphological alterations appearing on the erythrocyte membranes may be a manifestation of freezing damage.     

Signaling Pathway in the Osmotic Resistance Induced by Angiotensin II AT2 Receptor Activation in Human Erythrocytes

Background:Angiotensin II regulates blood volume via AT1 (AT1R) and AT2 (AT2R) receptors. As cell integrity is an important feature of mature erythrocyte, we sought to evaluate, in vitro, whether angiotensin II modulates resistance to hemolysis and the signaling pathway involved.Methods:Human blood samples were collected and hemolysis assay and angiotensin II signaling pathway profiling in erythrocytes were done.Results:Hemolysis assay created a hemolysis curve in presence of Ang II in several concentrations (10^-6 M, 10^-8 M, 10^-10 M, 10^-12 M). Angiotensin II demonstrated protective effect, both in osmotic stressed and physiological situations, by reducing hemolysis in NaCl 0.4% and 0.9%. By adding receptors antagonists (losartan, AT1R antagonist and PD 123319, AT2R antagonist) and/or signaling modulators for AMPK, Akt/PI3K, p38 and PKC we showed the protective effect was enhanced with losartan and abolished with PD 123319. Also, we showed activation of p38 as well as PI3K/Akt pathways in this system.Conclusion:Ang II protects human erythrocytes from hypo-osmotic conditions-induced hemolysis by activating AT2 receptors and triggering intracellular pathways.Key Words: Angiotensin II, Erythrocyte, Osmotic fragility, Signaling pathway     

On the Dynamics of Abrin Binding to Receptor Sites in Normal and Epstein Barr Virus Transformed Lymphocyte Cell Cultures

Abstract

The effects of treatment with varying doses of abrin, a D-galactose binding lectin, on DNA and protein synthesis of normal and Epstein Barr Virus transformed lymphocytes has been investigated. Activation, stimulation, and relative toxicity factor indices are studied, as well as possible relationships between DNA and protein synthesis rates, as measured by simultaneous tritiated thymidine (³H-TdR) and ¹⁴C-leucine uptake. Studies of two new indices, the metabolic self and cross coupling indices lead to the prediction that there are three morphologically distinct subpopulations of EBV-transformed lymphocytes with different abrin receptor site concentrations. This prediction is supported by SEM morphological differences. Using data on EBV-transformed lymphocyte cell density as a function of time and dose of abrin, one can demonstrate that the mean number of receptors bound/EBV-lymphocyte needed to exert a biological influence lies in the interval 59,264 receptors/cell to 370,040 receptors/cell. Using a simple packing model, one can demonstrate that a theoretical estimate places the number of binding sites between 57,600 receptors/cell and 360,000 receptors/cell.     

Changes in selected haematological parameters at early stages of the rainbow trout, Oncorhynchus mykiss, subjected to metal toxicants: arsenic, cadmium and mercury

Haematological responses (cellular alterations) induced 6 months post‐treatment by three toxic metals, arsenic, cadmium and mercury, are reported. Treatment was at early stages of rainbow trout, Oncorhynchus mykiss, development. Different blood parameters (haematocrit level, red cells/mm³, white cells/mm³ red/white cell ratio and the white blood cell differential count) were examined in association with a histological examination. Significant differences were observed in the total number of leucocytes and the number of lymphocytes between the lesion‐bearing fish of the arsenic and cadmium group and the healthy fish. Subsequently, the blood films showed atypical lymphocytes.     

三株交叉反应性的流感病毒HA抗体与H1N1流感病毒的致病机制分析

目的:结合临床甲型流感病例分析流感病毒可能的致病机理。方法:收集87份陕西省2009年甲型H1N1流感重症,危重症及死亡病例的血常规参数,对其淋巴细胞、红细胞和血小板三个指标分析。制备针对甲型H1N1的单克隆抗体,采用抗体亚类鉴定试剂盒分析其抗体轻链和重链的亚型,通过血凝活性实验检测三株抗体的血凝抑制活性,通过ELISA检测三株抗体与人和小鼠的血红蛋白、红细胞、白细胞膜和血小板膜的反应,通过免疫组化分析三株流感病毒抗体与正常小鼠肺组织的结合。结果:流感病毒感染后的死亡病例中淋巴细胞、红细胞和血小板均明显降低。三株抗体与人和小鼠的淋巴细胞、红细胞和血小板均有不同程度的交叉反应;免疫组化结果同时也证实三株HA抗体与小鼠的肺组织有不同的结合力。结论:流感病毒致病的原因可能与流感病毒感染机体后产生的抗体可与血液和组织中的成分结合有关。     

Differential resistance to calcium-induced bilirubin-dependent hemolysis in mammalian erythrocytes

Washed erythrocytes from human, buffalo, sheep and goat preincubated with different concentrations of calcium chloride (16.7–1830 μM) showed significantly different rates of hemolysis (up to 62%) after addition of bilirubin (72 μM). Goat erythrocytes displayed marked resistance to hemolysis with only 11% hemolysis observed at the highest calcium concentration. Similar trend in hemolysis was also observed when the concentration of CaCl₂ was fixed (330 μM) and bilirubin concentration varied (0–72 μM). (Ca²⁺–Mg²⁺)-ATPase levels were found significantly lower in goat and sheep erythrocyte membranes compared to human and buffalo erythrocyte membranes. This was correlated well with the observed hemolysis in various mammalian erythrocytes.     

Thermal instability of red blood cell membrane bilayers: Temperature dependence of hemolysis

Summary Rates of human red blood cell hemolysis were measured as a function of temperature. Three distinct temperature intervals for hemolysis were noted: a) At temperatures equal to or less than 37°C no hemolysis was observed for the duration of the incubation (30 hr). b) For temperatures exceeding 45°C hemolysis rates are rapid and are accompanied by gross changes in cellular morphology. The activation energy for hemolysis is 80 kcal/mole; this value is characteristic of protein denaturation and enzyme inactivation suggesting that these processes contribute to hemolysis at these high temperatures. c) Between 38 and 45°C the energy of activation is 29 kcal/mole, indicating that a fundamentally different process than protein inactivation is responsible for hemolysis at these relatively low temperatures. A mechanism based on the concept of the critical bilayer assembly temperature of cell membranes (N.L. Gershfeld,Biophys. J. 50:457–461, 1986) accounts for hemolysis at these relatively mild temperatures: The unilamellar state of the membrane is stable at 37°C, but is transformed to a multibilayer when the temperature is raised; hemolysis results because formation of the multibilayer requires exposing lipid-free areas of the erythrocyte surface. An analysis of the activation energy for hemolysis is presented that is consistent with the proposed unilamellar-multibilayer transformation.     

八种常见儿童感染性疾病相关指标差异性研究

摘要 目的：探讨异型淋巴细胞、白细胞、中性粒细胞、淋巴细胞、单核细胞、C-反应蛋白及血沉等在儿童感染性相关疾病不同疾病种类、不同年龄段及性别的差异情况,并进行分析,从而为疾病的诊断、鉴别诊断及治疗提供依据。方法：选取2017年1月-2018年12月266例患感染性疾病的儿童,根据疾病类型分为八组,分别为传染性单核细胞增多症（41例）、EB病毒感染（18例）、支气管肺炎（43例）、支气管炎（42例）、急性上呼吸道感染（48例）、急性化脓性扁桃体炎（18例）、肺炎（19例）、粒细胞减少（37例）等,应用检验相关手段,对患儿异型淋巴细胞、白细胞、中性粒细胞、淋巴细胞、单核细胞、C-反应蛋白及血沉等进行检测,采用SPSS17.0统计学分析软件,计算各检验结果的平均值与标准差,进行相关t检验,进而对不同疾病的检验项目进行统计学分析。结果：在八种疾病相关检验的比较中,传染性单核细胞增多症与其余七种感染性疾病在异型淋巴细胞、中性粒细胞、淋巴细胞及血沉的数据进行比较,其之间比较差异有统计学意义（P<0.05）,其余各组疾病相关指标没有统计学差异。在年龄段的分组比较中,异型淋巴细胞在各年龄段之间比较差异有统计学意义（P<0.05）。在性别分组比较中,白细胞在性别与各年龄段之间比较差异有统计学意义（P<0.05）。结论：不同种类感染性疾病及不同年龄、性别相关细胞检查存在差异,各类检测结果的综合对比分析,更有利于疾病的诊断及鉴别诊断。