scholarly journals The ultrastructure changes of Haemonchus contortus exposed to bamboo leaves (Gigantochloa apus) aqueous extract under in vitro condition

2020 ◽  
Vol 22 (1) ◽  
Author(s):  
Budi Purwo Widiarso ◽  
WISNU NURCAHYO ◽  
KURNIASIH KURNIASIH ◽  
JOKO PRASTOWO

Abstract. Widiarso, Nurcahyo W, Kurniasih, Prastowo J. 2021. The ultrastructure changes of Haemonchus contortus exposed to bamboo leaves (Gigantochloa apus) aqueous extract under in vitro condition. Biodiversitas 22: 1-5. The ultrastructural changes induced in adult Haemonchus contortus in vitro using the aqueous extract of bamboo leaves were assessed using scanning electron microscopy (SEM). The H. contortus adult females were obtained from three groups and treatment was repeated thrice. The first group (T0) was not treated with bamboo leaves; 100% of the worms lived. The second group (T1), treated with 0.1% bamboo leaf-extract, had 50% mortality 4 h after examination. The third group (T2), treated with 1% bamboo leaf-extract, had 100% mortality 4 h after examination. Five worms used per treatment were submerged in ethanol and incubated for 24 hours. The ultrastructural changes observed by SEM revealed structural alteration of the worm surface after in vitro contact with the bamboo leaf aqueous extract and compared to the control worms. The main changes concerned the anterior end or cephalic region, cuticle surface, and vulva flap area. The structural modification of the external part of the female reproductive system was found only in vitro. The structural changes found in the worms exposed to the bamboo leaves might affect their motility and nutrition with possible consequences on their reproduction. Transmission electron microscopy may help to understand the external changes observed in H. contortus.

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Mutsumi Yokota ◽  
Soichiro Kakuta ◽  
Takahiro Shiga ◽  
Kei-ichi Ishikawa ◽  
Hideyuki Okano ◽  
...  

AbstractMitochondrial structural changes are associated with the regulation of mitochondrial function, apoptosis, and neurodegenerative diseases. PRKN is known to be involved with various mechanisms of mitochondrial quality control including mitochondrial structural changes. Parkinson’s disease (PD) with PRKN mutations is characterized by the preferential degeneration of dopaminergic neurons in the substantia nigra pars compacta, which has been suggested to result from the accumulation of damaged mitochondria. However, ultrastructural changes of mitochondria specifically in dopaminergic neurons derived from iPSC have rarely been analyzed. The main reason for this would be that the dopaminergic neurons cannot be distinguished directly among a mixture of iPSC-derived differentiated cells under electron microscopy. To selectively label dopaminergic neurons and analyze mitochondrial morphology at the ultrastructural level, we generated control and PRKN-mutated patient tyrosine hydroxylase reporter (TH-GFP) induced pluripotent stem cell (iPSC) lines. Correlative light-electron microscopy analysis and live cell imaging of GFP-expressing dopaminergic neurons indicated that iPSC-derived dopaminergic neurons had smaller and less functional mitochondria than those in non-dopaminergic neurons. Furthermore, the formation of spheroid-shaped mitochondria, which was induced in control dopaminergic neurons by a mitochondrial uncoupler, was inhibited in the PRKN-mutated dopaminergic neurons. These results indicate that our established TH-GFP iPSC lines are useful for characterizing mitochondrial morphology, such as spheroid-shaped mitochondria, in dopaminergic neurons among a mixture of various cell types. Our in vitro model would provide insights into the vulnerability of dopaminergic neurons and the processes leading to the preferential loss of dopaminergic neurons in patients with PRKN mutations.


2016 ◽  
Vol 91 (4) ◽  
pp. 454-461 ◽  
Author(s):  
R.S. Tomar ◽  
S. Preet

AbstractThe present study focuses on the in vitro anthelmintic activity of silver nanoparticles (AgNPs) synthesized using the aqueous extract of Azadirachta indica against Haemonchus contortus. The synthesized AgNPs were characterized by ultraviolet–visible (UV-Vis) spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray diffraction (XRD) studies. The UV-Vis spectrum exhibited a sharp peak at 420 nm, which was validated by electron microscopy, indicating the preparation of spherical nanoparticles measuring 15–25 nm in size. The in vitro study was based on an egg hatch assay (EHA) and adult motility inhibition assays. Six concentrations of AgNPs were used for EHA, ranging from 0.00001 to 1.0 μg/ml, and a range of 1–25 μg/ml was used for adult worms. The highest concentration induced 85 ± 2.89% egg hatch inhibition. The IC50 value for EHA was 0.001 μg/ml, whereas in vitro adult H. contortus motility inhibition was produced at 7.89 μg/ml (LC50). The effectiveness of A. indica leaf extract (aqueous) was also evaluated, which showed an IC50 value for EHA of 115.67 μg/ml, while the LC50 against adult H. contortus was 588.54 μg/ml. The overall findings of the present study show that the experimental plant extract contains reducing properties for the synthesis of AgNPs which, in turn, showed potent anthelmintic properties. This is the first report where AgNPs have been tested for their anthelmintic properties in an in vitro model.


2015 ◽  
Vol 36 (6) ◽  
pp. 3763 ◽  
Author(s):  
Eidi Yoshihara ◽  
Alessandro Pelegrine Minho ◽  
Victor Bittencourt Dutra Tabacow ◽  
Sérgio Tosi Cardim ◽  
Milton Hissashi Yamamura

The parasite Haemonchus contortus is one of the most pathogenic for small ruminants in tropical and subtropical regions worldwide, including Brazil. The objective of this study was to evaluate the structural changes induced in adult H. contortus after in vitro contact with Acacia mearnsii extract (AE), using scanning electron microscopy. Adult nematodes were collected from a naturally infected lamb. In the in vitro assay the parasites were placed in contact with AE (100 mg ml-1), for two hours at 37oC. The nematodes used in the assays (exposed to AE and the negative controls) were analyzed using an electron scanning microscope (quadruplicate per treatment). In all replicates, similar morphological alterations were observed on the entire extension of the cuticle of the specimens that remained in contact with the EA in vitro assays, none significant lesion was observed in the negative control (not exposed to AE). These results indicate the direct action of EA on the cuticle of H. contortus in in vitro trials


2007 ◽  
Vol 19 (2) ◽  
pp. 383 ◽  
Author(s):  
Duangjai Boonkusol ◽  
Tassanee Faisaikarm ◽  
Andras Dinnyes ◽  
Yindee Kitiyanant

The purpose of the present study was to investigate the effects of two vitrification procedures on developmental capacity and ultrastructural changes of matured swamp buffalo oocytes. In vitro-matured oocytes were vitrified by using 35 and 40% ethylene glycol as vitrification solution for solid surface vitrification (SSV) and in-straw vitrification (ISV), respectively. Survival rate of vitrified–warmed oocytes, evaluated on the basis of ooplasm homogeneity, oolemma integrity and zona pellucida intactness, as well as parthenogenetic blastocyst rates of vitrified–warmed oocytes were significantly higher with SSV (89.3 and 13.6%, respectively) than ISV (81.8 and 5.5%, respectively). However, they were still significantly lower than that of control oocytes (100 and 34.2%, respectively). For examining the ultrastructural changes, fresh, VS-exposed (ISV and SSV), and vitrified–warmed (ISV and SSV) oocytes were processed for transmission electron microscopy. In VS-exposed oocytes, reduction of microvilli abundance and damage of mitochondrial membrane were found only in the ISV group. In vitrified–warmed oocytes, however, it was clear that both methods of vitrification induced profound ultrastructural modifications to microvilli, mitochondria, oolemma and cortical granules as well as to the size and position of vesicles. Damaged mitochondria were, however, more abundant in ISV vitrified oocytes than in SSV vitrified oocytes, which correlated with the developmental data, showing the superiority of the SSV method. The present study demonstrated the feasibility of vitrification of in vitro-matured swamp buffalo oocytes.


Neurosurgery ◽  
1989 ◽  
Vol 25 (5) ◽  
pp. 736-746 ◽  
Author(s):  
J. M. Findlay ◽  
B. K. A. Weir ◽  
K. Kanamaru ◽  
F. Espinosa

Abstract A right-sided subarachnoid hemorrhage (SAH) was created in 12 monkeys. Only the right (clot-side) cerebral arteries developed angiographic vasospasm (VSP), which was maximal 7 days after SAH. Eight animals were killed at this time and the remainder at 14 days. At the time of killing the middle cerebral arteries (MCAs) were harvested, and four normal, left (non-clot-side) MCAs were vasoconstricted in vitro with prostaglandin F2… All MCAs were studied with scanning and transmission electron microscopy. Right MCAs in maximal VSP 7 days from SAH were undistinguishable on scanning electron microscopy from normal arteries vasoconstricted in vitro: both groups demonstrated a mean 57% reduction in vessel caliber and a 5-fold increase in vessel wall thickness compared to normal, nonvasoconstricted left MCAs. On transmission electron microscopy, however, arteries in SAH-induced VSP showed degenerative changes in the tunica intima and media. These changes were still evident at 14 days. despite considerable resolution of VSP. These findings, as well as those from other pathological studies of animal and human cerebral arteries in VSP, suggest that the arterial narrowing and vessel wall thickening seen within several weeks of SAH is due primarily to medial contraction, but unlike simple vasoconstruction, is associated with degenerative ultrastructural changes in the endothelium and vascular smooth muscle cells which may denote a temporarily irreversible state.


Author(s):  
Tai-Te Chao ◽  
John Sullivan ◽  
Awtar Krishan

Maytansine, a novel ansa macrolide (1), has potent anti-tumor and antimitotic activity (2, 3). It blocks cell cycle traverse in mitosis with resultant accumulation of metaphase cells (4). Inhibition of brain tubulin polymerization in vitro by maytansine has also been reported (3). The C-mitotic effect of this drug is similar to that of the well known Vinca- alkaloids, vinblastine and vincristine. This study was carried out to examine the effects of maytansine on the cell cycle traverse and the fine struc- I ture of human lymphoblasts.Log-phase cultures of CCRF-CEM human lymphoblasts were exposed to maytansine concentrations from 10-6 M to 10-10 M for 18 hrs. Aliquots of cells were removed for cell cycle analysis by flow microfluorometry (FMF) (5) and also processed for transmission electron microscopy (TEM). FMF analysis of cells treated with 10-8 M maytansine showed a reduction in the number of G1 cells and a corresponding build-up of cells with G2/M DNA content.


Author(s):  
Ann M. Thomas ◽  
Virginia Shemeley

Those samples which swell rapidly when exposed to water are, at best, difficult to section for transmission electron microscopy. Some materials literally burst out of the embedding block with the first pass by the knife, and even the most rapid cutting cycle produces sections of limited value. Many ion exchange resins swell in water; some undergo irreversible structural changes when dried. We developed our embedding procedure to handle this type of sample, but it should be applicable to many materials that present similar sectioning difficulties.The purpose of our embedding procedure is to build up a cross-linking network throughout the sample, while it is in a water swollen state. Our procedure was suggested to us by the work of Rosenberg, where he mentioned the formation of a tridimensional structure by the polymerization of the GMA biproduct, triglycol dimethacrylate.


Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo

Human malignant trophoblast cells in continuous culture were incubated for 3 days in medium containing 1 mM N6-O2'-dibutyryl cyclic adenosine 3':5'-monophosphate (dibutyryl cyclic AMP) and 1 mM theophylline. The culture fluid was replenished daily. Stimulated cultures secreted many times more chorionic gonadotropin and estrogens than did control cultures in the absence of increased cellular proliferation. Scanning electron microscopy revealed remarkable surface changes of stimulated cells. Control cells (not stimulated) were smooth or provided with varying numbers of microvilli (Fig. 1). The latter, usually, were short and thin. The surface features of stimulated cells were considerably different. There was marked increase of microvilli which appeared elongated and thick. Many cells were covered with confluent polypoid projections (Fig. 2). Transmission electron microscopy demonstrated marked activity of cytoplasmic organelles. Mitochondria were increased in number and size; some giant forms with numerous cristae were observed.


Author(s):  
P. Moine ◽  
G. M. Michal ◽  
R. Sinclair

Premartensitic effects in near equiatomic TiNi have been pointed out by several authors(1-5). These include anomalous contrast in electron microscopy images (mottling, striations, etc. ),diffraction effects(diffuse streaks, extra reflections, etc.), a resistivity peak above Ms (temperature at which a perceptible amount of martensite is formed without applied stress). However the structural changes occuring in this temperature range are not well understood. The purpose of this study is to clarify these phenomena.


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