Silver-Silica nanoparticles induced dose-dependent modulation of histopathological, immunohistochemical, ultrastructural, proinflammatory, and immune status of broiler chickens.

Silver nanoparticles (AgNPs) are a powerful disinfectant, but little information is available on their potential use as a growth promoter and the safety margin of this. In this study, 480 1-day-old Cobb chicks were assigned to one control and three treated groups. The treated groups were supplemented with silver-doped silica nanoparticles (SiO2@AgNPs) at three dietary levels (8, 16, and 20 mg/kg diet) for 35 days. The results revealed no significant changes in the growth performance and oxidative parameters, and in most of the hematological and biochemical parameters among the control and treated groups. In contrast, dose-dependent adverse effects were exerted on the histopathological structure and immunohistochemical expression of CD45 in liver, kidneys, and lymphoid organs (spleen, bursa, and thymus). In addition, the relative weight of lymphoid organs and the serum levels of immunoglobulins M and G were significantly diminished. Moreover, the gene expression of proinflammatory cytokines (IL-ß1 and TNF-α) and the ultrastructural morphology in breast muscle showed significant dose-dependent alterations. It could be concluded that the dietary supplementation of SiO2@AgNPs at a level of 8 mg/kg diet or more has dose-dependent proinflammatory and immunosuppressive effects on broiler chickens.

Evaluation of the acute toxic response induced by triazophos to the non-target green algae Chlorella pyrenoidosa.

Residues of triazophos in aquatic ecosystems due to extensive use for controlling pests in agriculture has became worldwide concern, while the toxic response of triazophos on the non-target green algae in aquatic environment is not well studied. Therefore, the acute (96 h) toxic effects of 1 and 10 mg/L triazophos on green algae Chlorella pyrenoidosa were evaluated in present study. The results showed that the growth was notably inhibited when treated with triazophos and the 96 h-EC50 (median inhibition concentration) were 12.79 mg/L. The content of photosynthetic pigments (including chl a, chl b, total-chl and carotinoids) clearly decreased under two treatments after 48 h and 96 h with exception for the values at 48 h exposure in 1 mg/L treatment. In addition, the transcript abundance of photosynthesis-related genes (psbA, psbC and rbcL) showed obvious decrease in above two treatments after exposure 96 h to triazophos. In response to 10 mg/L triazophos treatment, the morphology of thylakoid chloroplast of algal cells were obviously damaged. It was also found that starch granules increased with down-regulation of atpB gene expression in 10 mg/L treatment, which suggests that triazophos may inhibit the energy metabolism of C. pyrenoidosa. Moreover, the algal growth inhibition was along with the increase of intracellular reactive oxygen species (ROS), activity of antioxidant enzymes and malondialdehyde content indicating oxidative damage and lipid peroxidation in the algal cells. Our findings reveal that triazophos has potential toxicity and environmental risks to one of the primary producers green algae.

The Anthelmintic Quassinoids Ailanthone and Bruceine a Induce Infertility in the Model Organism Caenorhabditis elegans by an Apoptosis-like Mechanism Induced in Gonadal and Spermathecal Tissues.

In continuation of the search for new anthelmintic natural products, the study at hand investigated the nematicidal effects of the two naturally occurring quassinoids ailanthone and bruceine A against the reproductive system of the model nematode Caenorhabditis elegans to pinpoint their anthelmintic mode of action by the application of various microscopic techniques. Differential Interference Contrast (DIC) and the epifluorescence microscopy experiments used in the presented study indicated the genotoxic effects of the tested quassinoids (c ailanthone = 50 µM, c bruceine A = 100 µM) against the nuclei of the investigated gonadal and spermathecal tissues, leaving other morphological key features such as enterocytes or body wall muscle cells unimpaired. In order to gain nanoscopic insight into the morphology of the gonads as well as the considerably smaller spermathecae of C. elegans, an innovative protocol of polyethylene glycol embedding, ultra-sectioning, acridine orange staining, tissue identification by epifluorescence, and subsequent AFM-based ultrastructural data acquisition was applied. This sequence allowed the facile and fast assessment of the impact of quassinoid treatment not only on the gonadal but also on the considerably smaller spermathecal tissues of C. elegans. These first-time ultrastructural investigations on C. elegans gonads and spermathecae by AFM led to the identification of specific quassinoid-induced alterations to the nuclei of the reproductive tissues (e.g., highly condensed chromatin, impaired nuclear membrane morphology, as well as altered nucleolus morphology), altogether implying an apoptosis-like effect of ailanthone and bruceine A on the reproductive tissues of C. elegans.

Neotropical Stomopogon (Diptera, Muscidae): new species, redescriptions and key to species.

Stomopogon Malloch (Diptera, Muscidae) is a Neotropical genus recorded from Argentina, Bolivia, Brazil, Chile and Peru. In this contribution, the genus is revised, and three new species are described, one from Bolivia (Murillo, La Paz), one from Brazil (Palmas, Paraná) and one from Peru (Wayqecha, Cuzco). The descriptions include color images, illustrations and ultrastructural morphology of the terminalia of the adult. We provide an identification key to the species of Stomopogon and an updated map with species' distributions. Stomopogon acuta (Malloch, 1934) is proposed as a new junior synonym of S. inculta Stein, 1911.

Reactivity of human labial glands in response to cevimeline treatment.

Among the pathologies affecting the salivary glands, the Sjögren's syndrome (SS), an autoimmune disease, causes progressive destruction of the glandular tissue. The effect of SS is particularly evident on the labial glands and the morphological analysis of these minor glands is considered useful for diagnosis. Cevimeline hydrochloride (SNI), a selective muscarinic agonist drug, is one of the elective treatments for the hyposalivation due to SS, acting not only on major salivary glands, but also on labial glands since their secretion is primarily under parasympathetic control. Aim of this study is to describe the morphology of human labial glands treated with SNI by light, transmission, and high-resolution scanning electron microscopy. Moreover, a morphometric analysis was applied to the light and transmission electron microscopy micrographs to obtain data that were then compared with analogous data collected on control and carbachol-treated labial glands. Following SNI administration, the mucous tubules exhibited enlarged lumina, which were filled with a dense mucous secretion. Occasionally, small broken debris of the cells were retrieved into the lumen. In the mucous secretory cells, some mucous droplets fused to form a large vacuole-like structure. Similarly, the seromucous acini showed both dilated lumina and canaliculi. These above reported signs of secretion were confirmed through morphometric analysis and a milder action of SNI than carbachol on labial parenchyma was observed. This study confirmed that SNI also evoked secretion on labial glands and that its effect is more physiologic than that of the pan-muscarinic agonists.

Age-associated hemogram and ultrastructural leukocyte morphology in Pygocentrus nattereri (Characiformes: Serrasalmidae) from the Brazilian Pantanal

Pygocentrus nattereri is a carnivorous fish widely distributed in the Brazilian wetland and occupies different river strata. Based on the sexual maturity, the standard length (SL) has been used to categorize age and physiological attributes. However, hematological parameters and their relationship with age and growth features are unknow. Here, the aim was to evaluate the hematological profile and leukocyte data of juvenile and adult specimens, associating it with SL. Specimens of both sexes were weighted, and SL were measured for classify as juveniles (SL ≤16.0 cm) and adults (SL ≥16.1 cm). Blood was collected to perform the hemogram, for leukocyte quantification and ultrastructural descriptions. In general, hemogram values and leukocyte, lymphocyte, and monocyte counts were high in juveniles. Hemogram variables were negatively correlated with SL. Thrombocyte and neutrophil counts were higher in adults than in juveniles. SL was correlated with leukogram variables. Ultrastructural analysis showed that granular leukocyte count in P. nattereri was similar another fishes. PAS-positive granular leukocytes are slightly smaller than neutrophils and are characterized by dense elongated and semi-lunar shapes in the cytoplasm. Our results suggest that age-based SL may be useful for the analysis of the relationship between P. nattereri health status and wetland aquatic environments.(AU)

Pygocentrus nattereri é um peixe carnívoro amplamente distribuído no pantanal brasileiro e ocupa diferentes estratos de rios. Com base na maturidade sexual, o comprimento padrão (CP) tem sido usado para categorizar a idade e os atributos fisiológicos. No entanto, os parâmetros hematológicos e sua relação com a idade e características de crescimento são desconhecidos. O objetivo foi avaliar o perfil hematológico e leucocitário de espécimes juvenis e adultos, associando-o ao CP. Espécimes de ambos os sexos foram pesados e os CPs foram medidos para classificação em juvenis (CP ≤16,0 cm) e adultos (CP ≥16,1 cm). Foi coletado sangue para realização do hemograma, perfil e morfologia dos leucócitos. Em geral, os valores do hemograma e as contagens de leucócitos, linfócitos e monócitos foram maiores nos jovens. A contagem de trombócitos e neutrófilos foi maior em adultos do que em jovens. A análise ultraestrutural mostrou que a contagem de leucócitos granulares em P. nattereri foi semelhante a outros peixes. Os leucócitos granulares PAS-positivos são menores do que os neutrófilos, sendo caracterizados por formas alongadas e semilunares densas no citoplasma. Nossos resultados sugerem que CP é útil para a análise da relação entre o estado de saúde de P. nattereri em ambientes aquáticos alagados.(AU)

Hyperglycemia alters lipid metabolism and ultrastructural morphology of cerebellum in brains of diabetic rats: Therapeutic potential of raffia palm (Raphia hookeri G. Mann & H. Wendl) wine.

The present study investigated the effect of raffia palm (Raphia hookeri) wine (RPW) on hyperglycemia-mediated lipid metabolites and pathways, functional chemistry and ultrastructural morphology of cerebellums in type 2 diabetes (T2D). T2D was induced in male Sprague-Dawley rats by feeding with 10% fructose ad libitum for 2 weeks before injecting intraperitoneally with 40 mg/kg bodyweight (bw) streptozotocin. Following confirmation of hyperglycemia at blood glucose >200 mg/dL, diabetic rats were treated with RPW at 150 and 300 mg/kg bw respectively. Metformin served as the standard drug. Negative and normal controls consisted of untreated diabetic and non-diabetic rats, respectively. After 5 weeks of treatment, the rats were humanely sacrificed, and their cerebellum excised from the harvested brains. GC-MS analysis revealed significant alterations in cerebellar lipid metabolites depicted by changes in unsaturated and saturated fatty acids, fatty - esters, alcohols, and amides, glycols and steroids on induction of T2D. Pathway enrichment analysis of the lipid metabolites revealed inactivation of arachidonic metabolic pathway following T2D induction. Treatment with both doses of RPW restored most of the metabolites, while reactivating arachidonic acid metabolism (high dose only). Low dose of RPW led to the activation of retinol metabolism. Both doses of RPW maintained cerebellar functional chemistry as revealed by FTIR analysis. TEM analysis revealed swollen mitochondria, depleted numbers of synaptic vesicles, and shrunk synaptic clefts following induction of T2D. These ultrastructural morphologies were improved in RPW-treated rats. These results portray the therapeutic potential of raffia palm wine in the management of neurodegenerative complications in T2D.

The end-stage failing human myocardium - Where changes in ultrastructure of human cardiac muscle cells do not appear to dictate clinical outcomes.

Heart failure is the end stage of cardiovascular abnormalities. Studies have primarily focused on the functional changes of cardiomyocytes in the failing heart from different animal models with very little information in the human condition. In addition little is known about the ultrastructural changes that proceed in cardiomyocytes in route to failure. The aim of this study was to examine the ultrastructural changes in the myocardium of human with end-stage heart failure. Left ventricular myocardial tissue samples from 7 patients with end-stage heart failure were examined with transmission and scanning electron microscopy. All heart failure patients were of New York Heart Association (NYHA) class III-IV. The data indicated normal three-dimensional arrangement of cardiac muscle cells in failing myocardium. The various organelles in cardiomyocytes including the nucleus, mitochondria, myofibrils, T-tubules and intercalated discs did not exhibit any remarkable morphological changes. We did observe the appearance of small membrane bound vesicles which appear to be associated with the intercalated discs. The nearly normal ultrastructure and arrangement of cardiomyocytes was remarkable in contrast to the dramatic clinical status of these patients in heart failure. These observations support the hypothesis, that there are no dramatic changes in the ultrastructure or three-dimensional architecture of cardiomyocytes in end-stage failing human myocardium.

Effects of imidazolium-based ionic liquids with different anions on wheat seedlings.

The effects of five imidazolium ionic liquids with different anions were studied in hydroponically grown wheat seedlings at concentrations of 50, 100, 150, 200, 250, and 300 mg L-1. The results showed that shoots and roots grew shorter and dry weight decreased with increasing concentrations of ionic liquids. Moreover, the antioxidant enzyme activities decreased and malondialdehyde (MDA) content was greater in the leaves of wheat seedlings subjected to ionic liquid (IL) treatments. The order of influence of ionic liquids on these indexes was [C4mim][TfO]> [C4mim][Cl]> [C4mim][BF4]> [C4mim][Lact]> [C4mim][Ala]. A transmission electron microscope (TEM) was used to observe leaf and root cellular structures, such as chloroplast, nucleus, mitochondria, and rough endoplasmic reticulum, in wheat exposed to ionic liquids at a concentration of 150 mg L-1. The results showed that the cellular structures of wheat were affected, and the degree of the effect of five ILs was consistent with the general trend of the measured indexes in this study. Ionic liquids influence the growth of plants by impeding growth, disrupting metabolic physiology and changing cellular structures. The degree of toxicity of imidazolium-based ionic liquids with different anions varies.

High Nitrogen Supply Induces Physiological Responsiveness to Long Photoperiod in Barley.

Photoperiod and nutrient nitrogen (N) supply influence the growth, development, and productivity of crops. This study examined the physiological, biochemical, and morpho-anatomical traits of NA5 and NA9, two barley cultivars with contrasting photoperiod lengths, under the combined treatment of photoperiod regime and N supply. Under long photoperiod, high N supply decreased net photosynthesis; decreased chlorophyll a and chlorophyll a/b; decreased ascorbate peroxidase (APX), catalase (CAT), and superoxide dismutase (SOD) activities; decreased ascorbate, glutathione, soluble protein, and soluble sugar; destroyed mesophyll cell integrity; and increased [Formula: see text], malondialdehyde, and proline in both NA5 and NA9. Under short photoperiod, high N content increased net photosynthesis; increased chlorophyll a and chlorophyll a/b; increased APX, CAT, and SOD activities; and increased antioxidants, soluble protein, and soluble sugar in NA9 but decreased the same parameters in NA5. These results indicated that N supply strongly affected photosynthetic capacity and the balance of reactive oxygen species in response to short and long photoperiod. High N supply enhanced the sensitivity of long-day barley to photoperiod change by inhibiting photosynthesis and decreasing antioxidant defense ability. High N mitigated the undesirable effects of shortened photoperiod in short-day barley. Therefore, the data from this study revealed that N status affects adaptation to photoperiod changes by maintaining redox homeostasis and photosynthetic capacity.

Direct electric current modifies important cellular aspects and ultrastructure features of Candida albicans yeasts: Influence of doses and polarities.

Available treatments against human fungal pathogens present high levels of resistance, motivating the development of new antifungal therapies. In this context, the present work aimed to analyze direct electric current (DC) antifungal action, using an in vitro apparatus equipped with platinum electrodes. Candida albicans yeast cells were submitted to three distinct conditions of DC treatment (anodic flow-AF; electroionic flow-EIF; and cathodic flow-CF), as well as different charges, ranging from 0.03 to 2.40 C. Our results indicated C. albicans presented distinct sensibility depending on the DC intensity and polarity applied. Both the colony-forming unit assay and the cytometry flow with propidium iodide indicated a drastic reduction on cellular viability after AF treatment with 0.15 C, while CF- and EIF-treated cells stayed alive when DC doses were increased up to 2.40 C. Additionally, transmission electron microscopy revealed important ultrastructural alterations in AF-treated yeasts, including cell structure disorganization, ruptures in plasmatic membrane, and cytoplasmic rarefaction. This work emphasizes the importance of physical parameters (polarity and doses) in cellular damage, and brings new evidence for using electrotherapy to treat C. albicans pathology process. Bioelectromagnetics. 38:95-108, 2017. © 2016 Wiley Periodicals, Inc.

Morphology and staining behavior of neutrophilic and eosinophilic granulocytes of the common marmoset (Callithrix jacchus).

Common marmosets (Callithrix jacchus) are frequently used as translational animal models for human diseases. However, a comparative study of cytological and histochemical detection methods as well as morphometric and ultrastructural characterization of neutrophils and eosinophils in this species is lacking. Blood samples of house dust mite sensitized and allergen challenged as well as lipopolysaccharide (LPS) challenged marmosets were analyzed with different cytological and histological staining methods. Furthermore, cell size and number of nuclear segments were compared between neutrophils and eosinophils. Electron microscopy was performed to characterize the ultrastructure of granulocytes. Of all applied cytological stains, three allowed differentiation of eosinophils and neutrophils and, thus, reliable quantification in blood smears: May-Grünwald-Giemsa stain, Congo Red and Naphthol AS-D Chloroacetate-Esterase. For histology, Hematoxylin-Eosin (H&E) could not demonstrate clear differences, whereas Sirius Red, Congo Red, and Naphthol AS-D Chloroacetate Esterase showed capable results for identification of eosinophils or neutrophils in lung tissue. Morphometry revealed that marmoset neutrophils have more nuclear segments and are slightly larger than eosinophils. Ultrastructurally, eosinophils presented with large homogeneous electron-dense granules without crystalloid cores, while neutrophils were characterized by heterogeneous granules of different size and density. Additionally, sombrero-like vesicles were detected in tissue eosinophils of atopic marmosets, indicative for hypersensitivity-related piecemeal degranulation. In conclusion, we provide a detailed overview of marmoset eosinophils and neutrophils, important for phenotypic characterization of marmoset models for human airway diseases.

Mouthparts in Leptotrombidium larvae (Acariformes: Trombiculidae).

Mouthparts of Leptotrombidium larvae (Acariformes: Trombiculidae), potential vectors of tsutsugamushi disease agents, were studied in detail using light microscopy, scanning electron microscopy, and transmission electron microscopy. The mouthparts incorporated within the pseudotagma gnathosoma are composed of the infracapitulum ventrally and the chelicerae dorsally. The ventral wall of the infracapitulum is formed by a wide mentum posteriorly and a narrowed malapophysis anteriorly. The malapophysis firmly envelops the distal cheliceral portions by its lateral walls. The lateral lips of the malapophysis are flexible structures hiding the cheliceral blades in inactive condition and turning back forming a type of temporary sucker closely applied to the host skin during feeding. The roof of the infracapitulum is formed by a weakly sclerotized labrum anteriorly and a cervix with the capitular apodemes extending posteriorly. The labral muscles are lacking. The capitular apodemes serve as origin for pharyngeal dilators running to the dorsal wall of the pharynx fused with the bottom of the infracapitulum. The basal cheliceral segments are separated from each other besides the very posterior portions where they are movably joined by the inner walls. The sigmoid pieces serve for insertion of the cheliceral elevators originating at the posterior portions of the basal segments. The movable digits reveal the solid basal sclerite and the cheliceral blade curved upward with a tricuspid cap on its tip. Dendrites of nerve cells run along the digits to their tips. The ganglia are placed within the basal segments just behind the movable digits. The chelicerae also reveal well developed flexible fixed digits overhanging the basal portions of the blades. The gnathosoma possesses several sets of extrinsic muscles originating at the scutum and at the soft cuticle behind it. Laterally, the gnathosoma bears five-segmented palps with a trifurcate palpal claw.

Phytotoxicity and oxidative stress effect of 1-octyl-3-methylimidazolium chloride ionic liquid on rice seedlings.

The effects of the ionic liquid 1-octyl-3-methylimidazolium chloride ([OMIM]Cl) was studied in hydroponically grown rice seedlings. Observed effects included increased root length and weight at concentrations of 0.1 mg/L [OMIM]Cl, however, as concentrations increased a reversed response was observed where roots and stems grew shorter and the weight decreased. The inhibitory concentration 50 (IC50,5d) values for root length and stem length were 0.59 mg/L and 0.70 mg/L, respectively. The Hill reaction activity and root system activity in [OMIM]Cl-treated rice seedlings were observed to be lower than the controls, however, root membrane permeability increased. The antioxidant enzyme activity in roots decreased, while in leaves there was an initial stimulation followed by a decrease. Malondialdehyde (MDA) content was found to be greater in seedlings subjected to [OMIM]Cl treatment. The cellular structures, such as chloroplasts, mitochondria and rough endoplasmic reticulum in rice root and leaf cells were affected at concentrations of 0.6 mg/L [OMIM]Cl.

Morphological Study of Respiratory Portion of Mouse Lung following Chemical Glue-Vapor Inhalation / 대한해부학회지

Industrial glues contain many kinds of organic solvents and glue sniffing by young people has become a social problem in Korea. Glue vapor may induce chronic toxicities different from those induced by exposures to the solvent of single component. We studied the effects of the inhalation of glue vapor on the primary target organ, the pulmonary epithelium of the respiratory system. Vapor samples of glue were collected for analysis; the components were acetone, n-hexane, methyl cyclopentane, c-hexane, and toluene. For the inhalation of glue vapor, experimental mice were exposed in a whole body chamber for 20 min/d for 3, 5, 7, and 14 d. Control groups were exposed to room air. Animals were euthanized and lung tissues were fixed in 10% neutral formalin for light microscopy, and in 2.5% glutaraldehyde plus 1.5% paraformaldehyde for electron microscopy. The results are as follows. 1. Alcianophilic bands were not detected in the normal alveolar epithelium, but weak alcianophilic bands were detected in bronchioles. Alcian blue-PAS and PAS positive cells were found in the mucosae of mice exposed to glue vapor for 5 and 7 d. 2. Types I and II pneumocytes and capillary endothelial cells were found in the normal alveolar epithelium. The blood-air barrier consists of Type I pneumocytes, a common basal lamina, and the capillary endothelium. 3. The alveolar epithelium of vapor-exposed mice showed more type II pneumocytes. In the longerexposed group, Type I pneumocytes and endothelial cells contained many pinocytotic vesicles. 4. The vapor-exposed lungs showed macrophages in the alveolar space, mild interstitial swelling, and increased numbers of collagenous fibers. Clearly, ultrastructural changes in pulmonary epithelia can occur following glue sniffing.