THE IN VITRO HATCHING OF ASCARIS LUMBRICOIDES EGGS

1961 ◽  
Vol 39 (2) ◽  
pp. 153-162 ◽  
Author(s):  
Donald Fairbairn
Keyword(s):  
Ascaris Lumbricoides ◽  
Morphological Changes ◽  
Potassium Dichromate ◽  
Vitelline Membrane ◽  
Potassium Dichromate Solution ◽  
Sodium Bisulphite ◽  
Reducing Conditions ◽  
Low Degree ◽  
First Results

The stimulus leading to in vitro hatching of Ascaris lumbricoides eggs consisted of four components as follows: (1) a temperature similar to that of homoiothermic animals, (2) pCO2 approximating 5 vol.%, (3) pH near 7.0, (4) non-specific reducing conditions such as are established by cysteine, glutathione, sodium hydrosulphite, sodium bisulphite, or sulphur dioxide. Application of this stimulus resulted in 80–95% hatching in 3 hours, the observed morphological changes corresponding to those occurring during intestinal hatching. The stimulus was not effective when applied to preinfective stages of embryonic development, or to infective eggs which were embryonated in 2% potassium dichromate solution. Maximal hatching was obtained when the stimulus was applied continuously. One of the first results of stimulation may be increased permeability of the vitelline membrane sufficient to permit the outward passage of enzymes such as chitinase which digest the chitinous shell. There is no reason to believe that the in vitro stimulus differs significantly from the natural stimulus, which is common to many homoiothermic animals and may account in part for the low degree of host-specificity in Ascaris infections.

scholarly journals Anticoccidial Activity of Aloe debrana and Aloe pulcherrima Leaf Gel against Eimeria Oocysts

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Andualem Yimer Desalegn ◽  
Mulubrihan Rahimeto Ahmed
Keyword(s):  
Infectious Diseases ◽  
Potassium Dichromate ◽  
Potassium Dichromate Solution ◽  
Control Groups ◽  
Development Of Resistance ◽  
Frequent Use ◽  
Huge Impact ◽  
Anticoccidial Activity ◽  
Avian Coccidiosis

Avian coccidiosis is one of the serious infectious diseases that pose huge impact on the health and production of poultry, hence mainly controlled by regular use of prophylactic and therapeutic chemical drugs. Frequent use of anticoccidial drugs, however, has resulted in the development of resistance in the Eimeria species and concerns about drug residues which have stimulated the efforts to search for alternative. Aloe pulcherrima and Aloe debrana are some of the endemic Aloe species of Ethiopia which are traditionally used for the treatment of various infectious diseases. In this study, an in vitro trial was undertaken to evaluate the effect of Aloe debrana and A. pulcherrima leaf gel infusions on the inhibition of the sporulation of oocysts of mixed Eimeria species isolated from naturally infected chickens. In this assay, petri dishes containing unsporulated coccidian oocysts at a dose of 1500 oocysts/ml of fecal solution were randomly assigned to 10, 15, 25, and 30% w/v crude gel infusion of both aloe species in 1% potassium dichromate solution while Amprolium and distilled water served as control groups. The results of this study show that 10, 15, 25, and 3 0% w/v gel infusions at the tested concentrations have anticoccidial activity as evidenced by their ability to decrease significantly (P<0.05) the sporulation of Eimeria oocysts relative to the control incubation. The efficacy of A. debrana was found significantly better (P<0.05) than A. pulcherrima at different concentrations. However, A. debrana at 30% concentration showed significantly higher (P<0.05) sporulation inhibition efficacy of 79.35% (CI: 75.99-83.21) compared to A. pulcherrima (69.17%, CI: 64.65-73.92) at similar concentration in relation to the control incubation, though this could not be compared to Amprolium which was more effective (P<0.05) with an inhibition percentage of 90.54% (CI: 89.16-92.21). This study has shown that there is potential for use of Aloe debrana leaf gel for the control of avian coccidiosis and as a chemotherapeutic, though much research is needed to determine absolute concentration which will make it comparable to commercially available drugs in terms of efficacy.

Resistance of faecal cysts of Spironucleus muris to some physical factors and chemical substances

1978 ◽  
Vol 12 (2) ◽  
pp. 95-97 ◽  
Author(s):  
I. Kunstýy̌ ◽  
E. Ammerpohl
Keyword(s):  
Osmotic Pressure ◽  
Chloride Solution ◽  
Room Temperature ◽  
Potassium Dichromate ◽  
Low Ph ◽  
Potassium Dichromate Solution ◽  
Physical Factors ◽  
High And Low Temperatures

The effect of 5 disinfectants, and of saturated zinc chloride solution + sodium chloride solution, 2·5% potassium dichromate solution, and 0·12% dimetridazole on faecal cysts of Spironuc1eus muris were tested in vitro. The resistance of the cysts to high and low temperatures, low pH, high osmotic pressure, centrifugation and desiccation was also tested. After treatment the morphology of the cysts was observed microscopically and their infectivity tested in vivo on sensitive thymus-deficient nude mice. The cysts ceased to be infective after treatment with most of usual distinfectants and by high temperature (45°C for 30 min). They resisted 0·12% dimetridazole, low temperature (-196°C), low pH (2·2), high osmotic pressure (distilled water and 30% 'Ficoll'), centrifugation (1500 g/20 min) and desiccation (room temperature for 14 days). These data may be useful for the control of Spironuc1eus muris infection in rodents and for cryopreservation of the parasite for experimental purposes.

scholarly journals In vitro anticoccidial activity of Vitis vinifera extract on oocysts of different Eimeria species of Broiler Chicken

2020 ◽  
Vol 71 (3) ◽  
pp. 2267
Author(s):  
R.Z. ABBAS ◽  
A. ABBAS ◽  
Z. IQBAL ◽  
M.A. RAZA ◽  
K. HUSSAIN ◽  
...  
Keyword(s):  
Vitis Vinifera ◽  
Potassium Dichromate ◽  
Eimeria Species ◽  
Grape Seed ◽  
Inhibitory Effect ◽  
Potassium Dichromate Solution ◽  
Dependent Manner ◽  
Control Groups ◽  
Dose Dependent

In the current experiment, the in vitro anticoccidial effect of Vitis venifera (grape seed) extract was evaluated. For this purpose, an in vitro sporulation inhbition assay was used. Collected oocysts of four Eimeria species (E. tenella, E. necatrix, E. brunetti and E. mitis) were exposed to six different concentrations (w/v) of Vitis vinifera extract (VVE) in 10% Dimethylsulphoxide solution (DMSO), while Dimethylsulphoxide (DMSO) and Potassium dichromate solution (K2Cr2O7) served as control groups. The results of the present study revealed that V. vinifera extract showed inhibitory effect on sporulation (%) and damage (%) of Eimeria oocysts in a dose dependent manner as compared to both control groups. V. vinifera extract also damaged the morhology of oocysts in terms of shape, size and number of sporocysts.

Correlation Between Cellular Functions and Morphological Changes of Human Trophoblast in Vitro

1975 ◽  
Vol 33 ◽  
pp. 600-601
Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo
Keyword(s):  
Electron Microscopy ◽  
Cellular Proliferation ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Transmission Electron ◽  
Marked Activity ◽  
Malignant Trophoblast

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.

Effect of Temperature, Velocity Gradient and I.V. Heparin on in Vitro Blood Coagulation and Platelet Aggregation

1967 ◽  
Vol 17 (01/02) ◽  
pp. 112-119 ◽  
Author(s):  
L Dintenfass ◽  
M. C Rozenberg
Keyword(s):  
Blood Coagulation ◽  
Velocity Gradient ◽  
Elevated Temperatures ◽  
Morphological Changes ◽  
Effect Of Temperature ◽  
Clotting Time ◽  
Progressive Change ◽  
Aggregation Of Platelets ◽  
Microscopic Techniques

SummaryA study of blood coagulation was carried out by observing changes in the blood viscosity of blood coagulating in the cone-in-cone viscometer. The clots were investigated by microscopic techniques.Immediately after blood is obtained by venepuncture, viscosity of blood remains constant for a certain “latent” period. The duration of this period depends not only on the intrinsic properties of the blood sample, but also on temperature and rate of shear used during blood storage. An increase of temperature decreases the clotting time ; also, an increase in the rate of shear decreases the clotting time.It is confirmed that morphological changes take place in blood coagula as a function of the velocity gradient at which such coagulation takes place. There is a progressive change from the red clot to white thrombus as the rates of shear increase. Aggregation of platelets increases as the rate of shear increases.This pattern is maintained with changes of temperature, although aggregation of platelets appears to be increased at elevated temperatures.Intravenously added heparin affects the clotting time and the aggregation of platelets in in vitro coagulation.

Cytotoxic Effect of the Combination of Gemcitabine and Atorvastatin Loaded in Microemulsion on the HCT116 Colon Cancer Cells

2017 ◽  
Vol 9 (2) ◽  
Author(s):  
Mayson H. Alkhatib ◽  
Dalal Al-Saedi ◽  
Wadiah S. Backer
Keyword(s):  
Colon Cancer ◽  
Cancer Cells ◽  
Anticancer Drugs ◽  
Therapeutic Potential ◽  
Morphological Changes ◽  
In Vitro Study ◽  
Colon Cancer Cells ◽  
Apoptosis Detection ◽  
Hct116 Cells

The combination of anticancer drugs in nanoparticles has great potential as a promising strategy to maximize efficacies by eradicating resistant, reduce the dosage of the drug and minimize toxicities on the normal cells. Gemcitabine (GEM), a nucleoside analogue, and atorvastatin (ATV), a cholesterol lowering agent, have shown anticancer effect with some limitations. The objective of this in vitro study was to evaluate the antitumor activity of the combination therapy of GEM and ATVencapsulated in a microemulsion (ME) formulation in the HCT116 colon cancer cells. The cytotoxicity and efficacy of the formulation were assessed by the 3- (4,5dimethylthiazole-2-yl)-2,5-diphyneltetrazolium bromide (MTT) assay. The mechanism of cell death was examined by observing the morphological changes of treated cells under light microscope, identifying apoptosis by using the ApopNexin apoptosis detection kit, and viewing the morphological changes in the chromatin structure stained with 4′,6-diamidino-2-phenylindole (DAPI) under the inverted fluorescence microscope. It has been found that reducing the concentration of GEM loaded on ME (GEM-ME) from 5μM to 1.67μM by combining it with 3.33μM of ATV in a ME formulation (GEM/2ATV-ME) has preserved the strong cytotoxicity of GEM-ME against HCT116 cells. The current study proved that formulating GEM with ATV in ME has improved the therapeutic potential of GEM and ATV as anticancer drugs.

The Anti-tumor Activity and Mechanisms of rLj-RGD3 on Human Laryngeal Squamous Carcinoma Hep2 Cells

2020 ◽  
Vol 19 (17) ◽  
pp. 2108-2119
Author(s):  
Yang Jin ◽  
Li Lv ◽  
Shu-Xiang Ning ◽  
Ji-Hong Wang ◽  
Rong Xiao
Keyword(s):  
Wound Healing ◽  
Western Blot ◽  
Morphological Changes ◽  
In Vivo Studies ◽  
Migration And Invasion ◽  
Tunel Staining ◽  
Dna Ladder ◽  
Western Blot Assay

Background: Laryngeal Squamous Cell Carcinoma (LSCC) is a malignant epithelial tumor with poor prognosis and its incidence rate increased recently. rLj-RGD3, a recombinant protein cloned from the buccal gland of Lampetra japonica, contains three RGD motifs that could bind to integrins on the tumor cells. Methods: MTT assay was used to detect the inhibitory rate of viability. Giemsa’s staining assay was used to observe the morphological changes of cells. Hoechst 33258 and TUNEL staining assay, DNA ladder assay were used to examine the apoptotic. Western blot assay was applied to detect the change of the integrin signal pathway. Wound-healing assay, migration, and invasion assay were used to detect the mobility of Hep2 cells. H&E staining assay was used to show the arrangement of the Hep2 cells in the solid tumor tissues. Results: In the present study, rLj-RGD3 was shown to inhibit the viability of LSCC Hep2 cells in vitro by inducing apoptosis with an IC50 of 1.23µM. Western blot showed that the apoptosis of Hep2 cells induced by rLj- RGD3 was dependent on the integrin-FAK-Akt pathway. Wound healing, transwells, and western blot assays in vitro showed that rLj-RGD3 suppressed the migration and invasion of Hep2 cells by integrin-FAKpaxillin/ PLC pathway which could also affect the cytoskeleton arrangement in Hep2 cells. In in vivo studies, rLj-RGD3 inhibited the growth, tumor volume, and weight, as well as disturbed the tissue structure of the solid tumors in xenograft models of BALB/c nude mice without reducing their body weights. Conclusion: hese results suggested that rLj-RGD3 is an effective and safe suppressor on the growth and metastasis of LSCC Hep2 cells from both in vitro and in vivo experiments. rLj-RGD3 might be expected to become a novel anti-tumor drug to treat LSCC patients in the near future.

Ability of PknA, a mycobacterial eukaryotic-type serine/threonine kinase, to transphosphorylate MurD, a ligase involved in the process of peptidoglycan biosynthesis

2008 ◽  
Vol 415 (1) ◽  
pp. 27-33 ◽  
Author(s):  
Meghna Thakur ◽  
Pradip K. Chakraborti
Keyword(s):  
Protein Kinases ◽  
Solid Phase ◽  
Morphological Changes ◽  
Binding Assays ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Peptidoglycan Biosynthesis ◽  
Vitro Binding ◽  
Solid Phase Binding

Eukaryotic-type serine/threonine protein kinases in bacteria have been implicated in controlling a host of cellular activities. PknA is one of eleven such protein kinases from Mycobacterium tuberculosis which regulates morphological changes associated with cell division. In the present study we provide the evidence for the ability of PknA to transphosphorylate mMurD (mycobacterial UDP-N-acetylmuramoyl-L-alanine:D-glutamate-ligase), the enzyme involved in peptidoglycan biosynthesis. Its co-expression in Escherichia coli along with PknA resulted in phosphorylation of mMurD. Consistent with these observations, results of the solid-phase binding assays revealed a high-affinity in vitro binding between the two proteins. Furthermore, overexpression of m-murD in Mycobacterium smegmatis yielded a phosphorylated protein. The results of the present study therefore point towards the possibility of mMurD being a substrate of PknA.

scholarly journals Individual Expression of Hepatitis A Virus 3C Protease Induces Ferroptosis in Human Cells In Vitro

2021 ◽  
Vol 22 (15) ◽  
pp. 7906
Author(s):  
Alexey A. Komissarov ◽  
Maria A. Karaseva ◽  
Marina P. Roschina ◽  
Andrey V. Shubin ◽  
Nataliya A. Lunina ◽  
...  
Keyword(s):  
Cell Death ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Proteolytic Enzymes ◽  
Morphological Changes ◽  
Human Cells ◽  
Mitochondrial Potential ◽  
3C Protease ◽  
Wide Range

Regulated cell death (RCD) is a fundamental process common to nearly all living beings and essential for the development and tissue homeostasis in animals and humans. A wide range of molecules can induce RCD, including a number of viral proteolytic enzymes. To date, numerous data indicate that picornaviral 3C proteases can induce RCD. In most reported cases, these proteases induce classical caspase-dependent apoptosis. In contrast, the human hepatitis A virus 3C protease (3Cpro) has recently been shown to cause caspase-independent cell death accompanied by previously undescribed features. Here, we expressed 3Cpro in HEK293, HeLa, and A549 human cell lines to characterize 3Cpro-induced cell death morphologically and biochemically using flow cytometry and fluorescence microscopy. We found that dead cells demonstrated necrosis-like morphological changes including permeabilization of the plasma membrane, loss of mitochondrial potential, as well as mitochondria and nuclei swelling. Additionally, we showed that 3Cpro-induced cell death was efficiently blocked by ferroptosis inhibitors and was accompanied by intense lipid peroxidation. Taken together, these results indicate that 3Cpro induces ferroptosis upon its individual expression in human cells. This is the first demonstration that a proteolytic enzyme can induce ferroptosis, the recently discovered and actively studied type of RCD.

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