Effects of trace metal elements on ultrastructural features of hepatopancreas of Armadillidium granulatum Brandt, 1833 (Crustacea,Isopoda)

This study was conducted to compare metals bioaccumulation in the terrestrial isopod Armadillidium granulatum collected from Ghar El Melh lagoon. We focused on recognizing the effects of trace elements on hepatopancreas functional role. To this end, isopod specimens were exposed for 3 weeks to sediments contaminated with cadmium, copper, zinc, mercury, and nickel. Three concentrations were used in duplicate for each experimental condition. At the end of the experiment, metal body burdens were determined using flame atomic absorption spectrometry. Results of the bioaccumulation factor (BAF) showed that the species A. granulatum was classified as a Cu macroconcentrator (BAF > 2) and a Zn deconcentrator (BAF < 2). Dose dependent morphological and histological changes were observed in the hepatopancreas cells using transmission electron microscopy. The predominant features were: microvillus border disruption, condensation of the cytoplasm with increasing endoplasmic reticulum, mitochondria, lysosomes and granules that accumulated metals in B and S cells. The number of lipid droplets decreased especially after Cd, Zn, Hg, and Ni treatments. This study demonstrated that the terrestrial isopod A. gramulatum could be a good indicator of soil metal contamination.     

Physiological and behavioral responses of Orchestia gammarellus (Amphipoda,Talitridae) towards trace elements contamination soil

Effects of trace elements on the hepatopancreas ultrastructure and on the locomotor activity rhythm were investigated in the amphipod talitrid Orchestia gammarellus collected from Bizerte lagoon banks situated in the north of Tunisia. Animals were exposed to a series of contaminated soil with different concentrations of cadmium and zinc. The locomotor activity rhythm was studied under constant darkness. Histopathological analysis showed that the trace elements, especially the Cd, induced significant changes in the morphology and in the ultrastructural organization of hepatopancreatic cells. The significant alterations obtained were dose dependent. Concerning the behavioral response, results revealed the presence of two components whatever the experimental conditions. In addition, a great inter-individual variability of the locomotor rhythm was observed. Patterns were in majority bimodal for the control individuals and became unimodal and multimodal when exposed under Cd and Zn, respectively. Furthermore, ultradian and circadian periods were determined. The circadian period lengthened after Zn exposure. In addition, the locomotor activity rhythm was more stable for control individuals. However, those exposed to Cd were less active.     

Morphostructural and immunohistochemical study for evaluation of nano‐TiO2 toxicity in Armadillo officinalis Duméril, 1816 (Crustacea,Isopoda, Oniscidea)

Invertebrates are precious organisms in order to study environmental pollution. In particular, they appear to be suitable as a bioindicator species for pioneer ecotoxicity studies on new xenobiotics such as nanoparticles. In fact, they are able to absorb nanomaterials scattered in the environment in different ways and it's known the compartmentalization of nano‐sized contaminants in selected tissues and intracellular organelles. Titanium dioxide represents the most used nanoparticulate, destined to become probably ubiquitous in the environment. Recently, some research has been published on the toxic potential of nano‐TiO₂ in several animal species. Among all invertebrates, Oniscidean Isopods are the only taxon of Crustaceans that has become completely terrestrial, known as excellent bioindicators and bioaccumulators. They have a digestive gland, the hepatopancreas, which is the location of election for the accumulation of pollutants. For this reason, they are considered efficient animal models to ecological studies. For this study, we collected Armadillo officinalis from Natural Oriented Reserve of “Vendicari” (Sicily, Italy), to evaluate the toxicity of titanium dioxide (TiO₂) on their hepatopancreas, after a short period of exposure. We conducted morphostructural and immunohistochemistry assays. The results suggested a great capacity of the species of bioaccumulation of nanoparticles in the hepatopancreas, where a strong positivity to the metallothioneins was highlighted. Our study confirms that Oniscidean Isopods, in particular Armadillo officinalis, proved to be an appropriate indicator of pollution in terrestrial ecosystems from nanoparticles.     

Light and electron microscopy study of the spermatheca of Eupholidoptera chabrieri bimucronata (Ramme, 1927) and Uromenus brevicollis trinacriae La Greca 1964 (Orthoptera: Tettigoniidae)

A study by both optical and electron microscopy has been carried out on the spermatheca of Eupholidoptera chabrieri bimucronata and Uromenus brevicollis trinacriae (Orthoptera: Tettigoniidae). In both the examined species, the spermatheca consists of a sac/kidney‐shaped seminal receptacle and a more or less tortuous spermathecal duct that opens into the common oviduct. The wall of both the organs consists of a pseudostratified epithelium surmounted by a cuticular intima; the latter is made up of a thicker endocuticle and an epicuticle. The epithelium shows two different cell types, irregularly arranged and with well differentiated functions: cuticle‐forming and gland cells. In both the species, the cuticle‐forming cells perform other functions, in addition to producing the cuticular intima. The gland cells never come in contact with the cuticular intima, have inside the reservoir a secretion whose appearance can diversify also in contiguous zones of the seminal receptacle. Based on our findings in both the species, the functions of the seminal receptacle would differ from those of the spermathecal duct. In the latter, some areas of the wall of the connecting tract show an activity of lysis, by contiguous epithelial cells, that could play a role in control and selection of spermatozoa. As for the feather‐shaped spermatodesms, similar in both the species, freeze‐fracture observations have shown that the acrosome of each spermatozoon regularly covers three‐quarters of the extension of the acrosome of the following spermatozoon. Finally, the significance of our findings, compared with what is known in literature, is discussed. Microsc. Res. Tech. 78:577–586, 2015. © 2015 Wiley Periodicals, Inc.     

Physiological and histopathological responses of Porcellio laevis (Isopoda,Crustacea) as indicators of metal trace element contamination

This study was designed to assess the impact of the mixture of cadmium (Cd) and zinc (Zn) on the bioaccumulation and the ultrastructural changes in the hepatopancreas of Porcellio laevis (Latreille, 1804) after 4 weeks of exposure to contaminated Quercus leaves under laboratory conditions. For each metal, four concentrations were used with four replicates for each concentration. Metal concentrations in the hepatopancreas and the rest of the body were determined using atomic absorption spectrometry. From the first week until the end of the experiment, a weight gain in P. laevis was observed particularly between the first and the end of exposure from 93.3 ± 18.22 mg fw to 105.22 ± 16.16 mg fw and from 106.4 ± 22.67 mg fw to 125.9 ± 23.9 mg fw for Mix1 and Mix4, respectively. Additionally, the determined metal trace elements (MTE) concentrations in the hepatopancreas were considerably higher compared to those in the rest of the body and seem to be dose‐dependent. Using transmission electron microscopy (TEM), some alterations were highlighted in the hepatopancreas. The main observed alterations were (a) the destruction of the microvilli border in a considerable portion of cells, (b) the increase of the lipid droplets with different shapes and sizes, (c) the increase in the number of the mitochondria, and (d) the appearance of TE in the form of B‐type granules. The obtained results confirmed the ability of P. laevis to deal with high amounts of MTE, suggesting its possible use in future soil's biomonitoring programs.     

Hyaluronic acid inhibited the upregulation of heat shock protein 70 in human chondrocytes from osteoarthritis and Kashin-Beck disease

This study aimed to investigate the effect of hyaluronic acid (HA) on the expression of heat shock protein70 (HSP70) in chondrocytes isolated from patients with osteoarthritis (OA) and Kashin-Beck disease (KBD). Thechondrocytes were collected from OA and KBD patients, and chondrocytes isolated from patients of accident injurieswere used as the control. The chondrocytes were treated with HA at different doses. HSP70 expression in chondrocytesat both mRNA and protein levels was tested by PCR and Western blot analysis. Compared with control, both mRNAand protein levels of HSP70 were higher in chondrocytes from KBD and OA. However, HA at the dose of 500 μg/mL significantly inhibited HSP70 expression levels in both KBD and OA groups (P < 0.05). In conclusion, HSP70 ishighly expressed in chondrocytes of patients of OA and KBD. HA intervention inhibits the upregulation of HSP70 inchondrocytes of OA and KBD patients and could be a promising agent for treatment of OA and KBD.     

Computerized morphometry of the cirrhotic liver: comparative analysis in primary biliary cirrhosis, alcoholic cirrhosis, and posthepatitic cirrhosis

Fibrosis and nodular regeneration are the hallmarks of liver cirrhosis. To assess the degree of fibrosis and the severity of the structural changes affecting parenchymal and extraparenchymal components in liver cirrhosis, a computerized morphometric model has been applied to liver specimens from patients undergoing liver transplantation for primary biliary cirrhosis, posthepatitic and alcoholic cirrhosis. Fifty-eight hepatectomy specimens from patients undergoing liver transplantation for cirrhosis were analyzed: 17 alcoholic, 28 posthepatitic (HBV-related and HCV-related cirrhosis), and 13 primary biliary cirrhoses. Liver specimens were fixed in 10% neutral-buffered formalin and embedded in paraffin. Sections were stained with chromotrope-aniline blue method and monoclonal antibodies against cytokeratin 7 and CD31. Volume fractions of parenchymal compartment and fibrosis were stereologically determined on the specimens stained with chromotrope-aniline blue method. Volume fractions of portal bile ducts, proliferated bile ductules, and hepatocytes with biliary metaplasia were measured on cytokeratin 7 stains, while volume fractions of capillary units have been evaluated on CD31 staining. Volume fraction of fibrosis was higher in primary biliary cirrhosis than in the other disease-induced cirrhosis. The main differences were related to immunohistochemical staining. Volume fraction of hepatocytes with biliary metaplasia was higher in HCV-related cirrhosis, whereas volume fractions of biliary structures were more prominent in HBV-related cirrhosis. Primary biliary cirrhosis was characterized by a reduced number of bile ducts and by a wider expression of cytokeratin 7 into periportal hepatocytes. Capillary units were more prominent in primary biliary cirrhosis than alcoholic and posthepatitic cirrhosis. Our computerized morphometric model well describes and quantifies the morphological alterations of the liver and it could represent an adjunctive tool to evaluate the degree of dysplastic phenomena involving parenchymal and extraparenchymal compartments.     

氢化物发生-原子荧光光谱法测定合金钢及合金中痕量元素铅、锡、砷、锑、铋的进展

通过对近年来有关氢化物发生-原子荧光光谱法在复杂基体体系中痕量元素检测方面的资料总结,阐述了氢化物发生-原子荧光光谱法分析过程中的干扰类型、干扰机理及消除干扰的措施,对其在合金钢及合金中痕量元素铅、锡、砷、锑、铋测定中的应用进展作了评述,并对氢化物发生-原子荧光光谱法的发展前景提出了展望。     

用同位素稀释电感耦合等离子体质谱法测定海水中的痕量铅

5 -磺基 -8-羟基喹啉用作微柱流动注射在线分离、富集和电感耦合等离子体质谱分析流程中的螯合剂 ,与铅反应生成铅螯合物被硅酸镁吸附剂吸附 ,实现痕量铅的分离、富集。对联用系统的参数如螯合剂用量、淋洗条件、洗脱条件和进样量等进行最优化 ,测定标准水样和加标海水的铅回收率分别为 1 0 1 %和 97.9% ,相对标准偏差为± 0 .98% ,检测限为 0 .2 0 4 μg/ L。可应用于生物、环境等高盐样品中铅的在线分离测定     

扫描电镜和X射线能谱仪对茶树菇子实体常见微量元素的定性定量分析

利用扫描电镜和 X射线能谱仪对茶树菇子实体的菌褶、菌柄、菌托和菌盖中常见的微量元素 Na、Mg、S、P、Ca、K、Fe、Cu、Mn和 Zn进行了测定 ,计算了它们的相对重量百分比。结果表明菌盖、菌托、菌褶和菌柄均含上述各种微量元素 ,但含量各不相同     

Detection of heat shock proteins 70 in the gill, liver, and cardiac muscle of Carassius auratus with confocal microscopy

Heat shock proteins 70 (Hsp70) are the most extensively studied heat shock proteins for the cellular abundance and cytoprotective effects. Hsp70 induction and subsequent quantification has been used as a sensitive system for aquatic toxicity risk assessment. In this study, the confocal microscopy was used to localize Hsp70 in Carassius auratus (C. auratus) with immunohistochemical technology. There are different zooms to select to analyze the object at the same field of vision with one objective lens with confocal microscopy. It need not change objective lens to observe the details of tissues. In this study, the tissue slices of C. auratus were observed with the 20-fold objective lens. Furthermore, the zooms of 1, 2, and 3 were used to acquire the distribution of Hsp70 in the tissue slices of C. auratus, and the clearer images of Hsp70 in the tissues were acquired. The results indicated that Hsp70 were present in the gill, liver, and cardiac muscle of C. auratus, and a method was established to detect Hsp70 in the tissues of C. auratus with confocal microscopy.     

Types of vascular wall as related to vasa vasorum in common tree shrew (Tupaia glis)

Blood vessels of the common tree shrew (Tupaia glis) were embedded in Araldite, sectioned at 0.5-1 microm thickness, and observed with light microscope (LM). It was found that the vascular wall could be classified into three categories: (1) those with proper vasa vasorum (PVV); (2) those with collaborative vasa vasorum in perimural tissues; (3) those without vasa vasorum. The PVV were located in the the tunica media, between the tunica media and the tunica adventitia, and in the tunica adventitia of the vascular wall. On studying the vascular casts with a scanning electron microscope (SEM), it was noted that what appeared to be vasa vasorum did not always conform to those found under the LM. Furthermore, the PVV seen in a particular blood vessel among different tree shrews were not always related to intraluminal partial pressure of oxygen, vascular size, and thickness of the wall. In contrast, the results of this study indicate that the activeness of the vascular wall in varying the amount of blood flow to certain organs is an important factor that is associated with the existence, density, and distribution of the vasa vasorum.     

液相色谱-串联质谱法测定猪肝中痕量己烯雌酚、己烷雌酚和双烯雌酚

建立了液相色谱-串联质谱法测定猪肝中痕量己烯雌酚、己烷雌酚和双烯雌酚。以氟化铵为流动相添加剂,目标物响应值平均提高约10倍,向样品中加入1%(V/V)乙酸乙腈超声提取,提取液经分散固相萃取净化、水稀释后,采用Waters Acquity UPLC HSS T3色谱柱,以0.1 mmol/L氟化铵和乙腈为流动相,梯度洗脱分离,电喷雾电离源负离子模式和多反应监测模式检测,外标法定量。在0.4、0.8、4.0μg/kg添加水平下,3种目标物的加标回收率为89.5%～109.4%,相对标准偏差(RSD)为2.2%～7.8%,方法定量限为0.4μg/kg。本方法前处理过程无需浓缩,操作简便、灵敏度高、重现性好、环保节约,适用于猪肝中痕量己烯雌酚、己烷雌酚和双烯雌酚的定性与定量分析。     

ICP-AES 法测定生活饮用水中铅、钡、钼等八种微量元素

通过实验,制定用ＩＣＰ－ＡＥＳ法直接测定生活饮用水中铅、钡、钼、硒、硼镍、镉、钒八种微量元素的企业标准方法。利用国家标准物质验证了方法的准确度,测定值与标准值吻合,相对标准偏差小于２．００％,水样加标回收率为９３．４％－１０６．４％,该方法操作简便、快速、准确、可靠、检出限低。