scholarly journals F127/Cisplatin Microemulsions: In Vitro, In Vivo and Computational Studies

2021 ◽  
Vol 11 (7) ◽  
pp. 3006
Author(s):  
Saman Sargazi ◽  
Mohammad Reza Hajinezhad ◽  
Mahmood Barani ◽  
Mahwash Mukhtar ◽  
Abbas Rahdar ◽  
...  
Keyword(s):  
Liver Enzymes ◽  
Colorimetric Assay ◽  
Morphological Changes ◽  
Effective Strategies ◽  
In Vivo Experiments ◽  
Mtt Colorimetric Assay ◽  
Male Wistar Rats

The development of effective strategies for local administration of chemotherapeutic drugs, thus minimizing the adverse side effects to patients, is one of the key challenges in biomedicine and cancer research. This work reports the formulation and characterization of PluronicF127 microemulsions to enhance the bioavailability of Cisplatin (Cis). The size of Cis microemulsion was about 12.0 nm, as assessed by dynamic light scattering analysis. In vitro cytotoxic activity of free Cis and F127/Cis microemulsions were studied on malignant (C152 and MCF7) and normal (HUVEC) cells via tetrazolium (MTT) colorimetric assay. Cell morphology was also monitored. In vitro assessments revealed thatF127/Cis microemulsions induced cytotoxicity/morphological changes to a lesser extent than free Cis. Regarding in vivo experiments, F127/Cis microemulsions were injected intraperitoneally at 7 and 14 mg/kg doses into adult male Wistar rats to assess histologic and biochemical changes. In this case, the bulk Cis group caused severe histopathological changes and significant increases in serum liver enzymes and serum kidney function markers. The group treated with the 14 mg/kg dose of F127/Cis microemulsions also showed severe fatty changes and significant increases in serum liver enzymes, blood urea nitrogen, and creatinine levels. The group treated with the low dose of nano-Cis showed a significant increase in serum liver enzymes levels accompanied by mild fatty changes of the liver. Theoretical surveys were performed to get an understanding of the interplay between F127 and Cis. Results reveal that hydrogen bonding (HB) interactions with F127have an influence on the molecular properties of Cis and may playa role in the lower toxicity of F127/Cis in comparison to free Cis.

scholarly journals Enlisting the Ixodes scapularis Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick—Pathogen Interactions

Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 70
Author(s):  
Lourdes Mateos-Hernández ◽  
Natália Pipová ◽  
Eléonore Allain ◽  
Céline Henry ◽  
Clotilde Rouxel ◽  
...  
Keyword(s):  
Cell Line ◽  
Peripheral Nerves ◽  
Ixodes Scapularis ◽  
Embryonic Cell ◽  
Cell Subpopulation ◽  
In Vivo Experiments

Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like Ixodes scapularis embryonic cell line, ISE6, is an effective tool frequently used for examining tick–pathogen interactions. We detected 37 neuropeptide transcripts in the I. scapularis ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, Anaplasma phagocytophilum. The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in A. phagocytophilum-infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of I. scapularis synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete Ixodes neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick–pathogen interactions.

scholarly journals Evaluation of Radiolabeled Girentuximab In Vitro and In Vivo

2018 ◽  
Vol 11 (4) ◽  
pp. 132 ◽  
Author(s):  
Tais Basaco ◽  
Stefanie Pektor ◽  
Josue Bermudez ◽  
Niurka Meneses ◽  
Manfred Heller ◽  
...  
Keyword(s):  
Specific Binding ◽  
Sodium Ascorbate ◽  
Tumor Uptake ◽  
Tetraacetic Acid ◽  
In Vivo Experiments ◽  
Caix Expression ◽  
Tetraazacyclododecane Tetraacetic Acid

Girentuximab (cG250) targets carbonic anhydrase IX (CAIX), a protein which is expressed on the surface of most renal cancer cells (RCCs). cG250 labeled with 177Lu has been used in clinical trials for radioimmunotherapy (RIT) of RCCs. In this work, an extensive characterization of the immunoconjugates allowed optimization of the labeling conditions with 177Lu while maintaining immunoreactivity of cG250, which was then investigated in in vitro and in vivo experiments. cG250 was conjugated with S-2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane tetraacetic acid (DOTA(SCN)) by using incubation times between 30 and 90 min and characterized by mass spectrometry. Immunoconjugates with five to ten DOTA(SCN) molecules per cG250 molecule were obtained. Conjugates with ratios less than six DOTA(SCN)/cG250 had higher in vitro antigen affinity, both pre- and postlabeling with 177Lu. Radiochemical stability increased, in the presence of sodium ascorbate, which prevents radiolysis. The immunoreactivity of the radiolabeled cG250 tested by specific binding to SK-RC-52 cells decreased when the DOTA content per conjugate increased. The in vivo tumor uptake was < 10% ID/g and independent of the total amount of protein in the range between 5 and 100 µg cG250 per animal. Low tumor uptake was found to be due to significant necrotic areas and heterogeneous CAIX expression. In addition, low vascularity indicated relatively poor accessibility of the CAIX target.

scholarly journals Characterization of Endogenous Retroviruses in Sheep

2003 ◽  
Vol 77 (20) ◽  
pp. 11268-11273 ◽  
Author(s):  
Nikolai Klymiuk ◽  
Mathias Müller ◽  
Gottfried Brem ◽  
Bernhard Aigner
Keyword(s):  
Endogenous Retrovirus ◽  
Ovis Aries ◽  
Endogenous Retroviruses ◽  
Open Reading Frames ◽  
In Vivo Experiments ◽  
Pregnant Sheep ◽  
Reading Frames

ABSTRACT Endogenous retrovirus (ERV) sequences have been found in all mammals. In vitro and in vivo experiments revealed ERV activation and cross-species infection in several species. Sheep (Ovis aries) are used for various biotechnological purposes; however, they have not yet been comprehensively screened for ERV sequences. Therefore, the aim of the study was to classify the ERV sequences in the ovine genome (OERV) by analyzing the retroviral pro-pol sequences. Three OERV β families and nine OERV γ families were revealed. Novel open reading frames (ORF) in the amplified proviral fragment were found in one OERV β family and two OERV γ families. Hybrid OERV produced by putative recombination events were not detected. Quantitative analysis of the OERV sequences in the ovine genome revealed no relevant variations in the endogenous retroviral loads of different breeds. Expression analysis of different tissues from fetal and pregnant sheep detected mRNA from both gammaretrovirus families, showing ORF fragments. Thus, the release of retroviruses from sheep cells cannot be excluded.

scholarly journals The Role of Thermal Effects in Plasma Medical Applications: Biological and Calorimetric Analysis

2019 ◽  
Vol 9 (24) ◽  
pp. 5560 ◽  
Author(s):  
Luigi Cordaro ◽  
Gianluca De Masi ◽  
Alessandro Fassina ◽  
Clarice Gareri ◽  
Antonio Pimazzoni ◽  
...  
Keyword(s):  
Thermal Effects ◽  
Plasma Source ◽  
Total Power ◽  
Therapeutic Effects ◽  
Calorimetric Analysis ◽  
Blood Samples ◽  
In Vivo Experiments ◽  
Male Wistar Rats

Plasma Medicine tools exploit the therapeutic effects of the exposure of living matter to plasma produced at atmospheric pressure. Since these plasmas are usually characterized by a non-thermal equilibrium (highly energetic electrons, low temperature ions), thermal effects on the substrate are usually considered negligible. Conversely, reactive oxygen and nitrogen species (RONS), UV radiation and metastables are thought to play a major role. In this contribution, we compare the presence of thermal effects in different operational regimes (corresponding to different power levels) of the Plasma Coagulation Controller (PCC), a plasma source specifically designed for accelerating blood coagulation. In particular, we analyze the application of PCC on human blood samples (in vitro) and male Wistar rats tissues (in vivo). Histological analysis points out, for the highest applied power regime, the onset of detrimental thermal effects such as red cell lysis in blood samples and tissues damages in in-vivo experiments. Calorimetric bench tests performed on metallic targets show that the current coupled by the plasma on the substrate induces most of measured thermal loads through a resistive coupling. Furthermore, the distance between the PCC nozzle and the target is found to strongly affect the total power.

scholarly journals A primary study on genes with selected mutations by in vitro passage of Chlamydia muridarum strains

2019 ◽  
Vol 77 (3) ◽  
Author(s):  
Zhou Zhou ◽  
Na Liu ◽  
Yingzi Wang ◽  
Arthur Wirekoh Emmanuel ◽  
Xiaoxing You ◽  
...  
Keyword(s):  
Single Gene ◽  
Point Mutations ◽  
Primary Study ◽  
Chlamydia Muridarum ◽  
In Vivo Experiments ◽  
High Incidence ◽  
Identification And Characterization

ABSTRACTObjectiveThis study is to investigate the functions of newly discovered genes in Chlamydia muridarum (C. muridarum) strains with single gene differences.MethodsUsing whole genome sequencing and plaque formation assays, C. muridarum parental and passaging strains were established, and the isogenic clones expressing certain genotypes were isolated. Strains with single gene differences were obtained. Based on prediction, the valuable strains with single gene differences of tc0412, tc0668 or tc0237 were subjected to the in vitro and in vivo experiments for biological characterization and virulence analysis.ResultsInsertional -472840T mutation of the tc0412 gene (T28T/B3 type) matching with the nonmutant tc0668 gene and tc0237 gene with point mutations G797659T (Q117E) might slow the growth of Chlamydia due to the lack of a plasmid. The nonmutant tc0668 in the strain might induce a high incidence of hydrosalpinx in mice, while tc0668 with a G797659T point mutation was significantly attenuated. Compared with the nonmutant tc0237, the strains containing mutant tc0237 were characterized by reduced centrifugation dependence during infection.ConclusionThe identification and characterization of these genes might contribute to the comprehensive understanding of the pathogenic mechanism of Chlamydia.

Emerging Vascular Applications of Magnetic Resonance Imaging: A Picture is Worth More than a Thousand Words

Vascular ◽  
2006 ◽  
Vol 14 (6) ◽  
pp. 366-371 ◽  
Author(s):  
Tamara N. Fitzgerald ◽  
Akihito Muto ◽  
Fabio Akimaro Kudo ◽  
Jose Mario Pimiento ◽  
Robert Todd Constable ◽  
...  
Keyword(s):  
Magnetic Resonance ◽  
Three Dimensional ◽  
Quantitative Information ◽  
Data Set ◽  
Vascular Intervention ◽  
In Vivo Experiments ◽  
Blood Flow Dynamics

Vascular applications of magnetic resonance (MR) imaging are reviewed, with emphasis on algorithms that use nonpictorial information contained in the MR data set. Current clinical vascular practice generally limits use of MR angiography and three-dimensional vessel images to qualitative pictorial rendering without routinely using the available quantitative information contained within the MR data. This review is dedicated to recent advances that include characterization of vessel histology, assessment of carotid plaque vulnerability, characterization of blood flow dynamics, quantitative analysis of disease severity, and prediction of vascular intervention outcome. Examples from histologic preparation, in vitro and in vivo experiments, are discussed, with an emphasis on potential clinical applications and advances in acquisition technology.

scholarly journals Antiviral Activity of Total Flavonoid Extracts fromSelaginella moellendorffiiHieron against Coxsackie Virus B3In VitroandIn Vivo

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Dan Yin ◽  
Juan Li ◽  
Xiang Lei ◽  
Yimei Liu ◽  
Zhanqiu Yang ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Inhibitory Concentration ◽  
Colorimetric Assay ◽  
Total Flavonoid ◽  
Coxsackie Virus ◽  
Mtt Colorimetric Assay ◽  
Diphenyl Tetrazolium Bromide ◽  
Selaginella Moellendorffii

The antiviral activity of total flavonoid extracts fromSelaginella moellendorffiiHieron and its main constituents amentoflavone were investigated against coxsackie virus B3 (CVB3). When added during or after viral infection, the extracts and amentoflavone prevented the cytopathic effect (CPE) of CVB3, as demonstrated in a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay, with a 50% inhibitory concentration (IC50) from19±1.6to41±1.2 μg/mL and25±1.2to52±0.8 μg/mL, respectively. KM mice were used as animal models to test the extracts' activityin vivo. Oral administration of the total flavonoid extracts at 300 mg/kg/day significantly reduced mean viral titers in the heart and kidneys as well as mortality after infection for 15 days. The experimental results demonstrate thatin vitroandin vivothe model mice infected with CVB3 can be effectively treated by the total flavonoid extracts fromSelaginella moellendorffiiHieron.

Leptin and Gaba Interactions on Thermoregulation of Rats

2014 ◽  
Vol 7 (1) ◽  
pp. 20-24 ◽  
Author(s):  
Krassimira S. Yakimova ◽  
Rumen P. Nikolov ◽  
Ivan G. Todorov ◽  
Milen H. Hristov
Keyword(s):  
Body Temperature ◽  
Core Body Temperature ◽  
Point Of View ◽  
The Body ◽  
In Vitro Experiments ◽  
In Vivo Experiments ◽  
Male Wistar Rats ◽  
In Vivo Effects

Abstract Leptin inhibits feeding, reduces body weight and increases thermogenesis. Experimental data suggest involvement of GABAergic mechanisms in the regulation of feeding behavior and energy balance. The present study was set to determine the effect of combinations from leptin, GABAB-agonist baclofen and GABAB-antagonist CGP35348 on thermoregulation of male Wistar rats, using in vivo and in vitro experiments. The substances used for in vivo experiments were administered intraperitoneally (i.p.). The measurement of the body temperature was done via thermistor probes (TX8) and monitored on multichannel recorder Iso-Thermex16. In vitro experiments were conducted on rat PO/AH neurons, recorded extracellulary by conventional electrophysiological equipment, using brain slice preparations. The separate intraperitoneal injection of leptin as well as GABAB-antagonist CGP35348 produced significant hyperthermia in rats while the GABAB-agonist baclofen caused a decrease in the core body temperature. The probable synergy between the hyperthermic effects of leptin and GABAB-antagonist did not occur. On the contrary, the effect of this combination was lower as compared to the result of the separate administration of GABAB-antagonist. When leptin was applied just prior to GABAB-agonist baclofen, neither of their separate effects appeared. In vivo effects determined correlated with in vitro changes of firing rate observed in PO/AH neurons. The data from this study provide a new point of view concerning the interactions of leptin and GABA on the level of thermoregulation. These results represent a step forward in understanding the complicated mechanisms involved in thermoregulation.

scholarly journals Discovery and characterization of a fourth class of guanidine riboswitches

2020 ◽  
Vol 48 (22) ◽  
pp. 12889-12899
Author(s):  
Felina Lenkeit ◽  
Iris Eckert ◽  
Jörg S Hartig ◽  
Zasha Weinberg
Keyword(s):  
Biological Significance ◽  
Protein Domains ◽  
Structural Class ◽  
New Class ◽  
In Vivo Experiments ◽  
Fourth Class ◽  
Metabolite Binding

Abstract Riboswitches are RNAs that specifically sense a small molecule and regulate genes accordingly. The recent discovery of guanidine-binding riboswitches revealed the biological significance of this compound, and uncovered genes related to its biology. For example, certain sugE genes encode guanidine exporters and are activated by the riboswitches to reduce toxic levels of guanidine in the cell. In order to study guanidine biology and riboswitches, we applied a bioinformatics strategy for discovering additional guanidine riboswitches by searching for new candidate motifs associated with sugE genes. Based on in vitro and in vivo experiments, we determined that one of our six best candidates is a new structural class of guanidine riboswitches. The expression of a genetic reporter was induced 80-fold in response to addition of 5 mM guanidine in Staphylococcus aureus. This new class, called the guanidine-IV riboswitch, reveals additional guanidine-associated protein domains that are extremely rarely or never associated with previously established guanidine riboswitches. Among these protein domains are two transporter families that are structurally distinct from SugE, and could represent novel types of guanidine exporters. These results establish a new metabolite-binding RNA, further validate a bioinformatics method for finding riboswitches and suggest substrate specificities for as-yet uncharacterized transporter proteins.

scholarly journals Virulence of Leishmania infantum Is Expressed as a Clonal and Dominant Phenotype in Experimental Infections

2001 ◽  
Vol 69 (12) ◽  
pp. 7365-7373 ◽  
Author(s):  
Yves Jean-François Garin ◽  
Annie Sulahian ◽  
Francine Pratlong ◽  
Pascale Meneceur ◽  
Jean-Pierre Gangneux ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Virulent Strain ◽  
Biological Marker ◽  
In Vivo Experiments ◽  
Dominant Phenotype ◽  
Virulence Phenotype ◽  
Parasite Populations

ABSTRACT Human Leishmania infantum infection results in a spectrum of clinical expressions ranging from cutaneous to either asymptomatic or fatal visceral disease. In this context, characterization of parasite virulence appears to be relevant as a biological marker of intrinsic parasitic factors that can affect the pathology of leishmaniasis. Since parasite populations in naturally infected hosts are likely to be composed of multiclonal associations, we first explored the biodiversity of parasite virulence at the intrastrain level in vitro and in vivo by using 11 clones isolated from three strains previously known to express different virulence phenotypes in mice. Subsequently, we studied the course of infection in mice inoculated simultaneously or successively with strains or clones showing various virulence phenotypes. Analysis of in vitro growth characteristics showed no differences among clones from the different parental strains. By contrast, in vivo experiments evidenced a marked intrastrain heterogeneity of virulence to mice. One out of five clones obtained from a virulent strain showed a typical virulence phenotype, while the remaining four clones had low-virulence profiles, as did the six clones isolated from two low-virulence strains. In mixed multiclonal infections, the virulence phenotype was expressed as a dominant character over the associated low-virulence clones. After a challenge with either a homologous or a heterologous strain or clone, virulence phenotypes were conserved and expressed as in naive mice independently from the preexisting population. These results strongly suggest that parasite virulence in L. infantum visceral leishmaniasis is clonal and dominant in nature.

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