scholarly journals Isolation and Characterization of a Novel Pathogenic Strain of Ehrlichia minasensis

2019 ◽  
Vol 7 (11) ◽  
pp. 528 ◽  
Author(s):  
Daniel Moura de Aguiar ◽  
João Pessoa Araújo Junior ◽  
Luciano Nakazato ◽  
Emilie Bard ◽  
Lisandra Aguilar-Bultet ◽  
...  
Keyword(s):  
Cell Line ◽  
Ixodes Scapularis ◽  
Hematological Parameters ◽  
Light And Electron Microscopy ◽  
Clinical Signs ◽  
Wide Distribution ◽  
Embryonic Cell ◽  
Holstein Breed ◽  
Isolation And Characterization

The genus Ehrlichia is composed of tick-borne obligate intracellular gram-negative alphaproteobacteria of the family Anaplasmataceae. Ehrlichia includes important pathogens affecting canids (E. canis, E. chaffeensis, and E. ewingii), rodents (E. muris), and ruminants (E. ruminantium). Ehrlichia minasensis, an Ehrlichia closely related to E. canis, was initially reported in Canada and Brazil. This bacterium has now been reported in Pakistan, Malaysia, China, Ethiopia, South Africa, and the Mediterranean island of Corsica, suggesting that E. minasensis has a wide geographical distribution. Previously, E. minasensis was found to cause clinical ehrlichiosis in an experimentally infected calf. The type strain E. minasensis UFMG-EV was successfully isolated from Rhipicephalus microplus ticks and propagated in the tick embryonic cell line of Ixodes scapularis (IDE8). However, the isolation and propagation of E. minasensis strains from cattle has remained elusive. In this study, the E. minasensis strain Cuiabá was isolated from an eight-month-old male calf of Holstein breed that was naturally infected with the bacterium. The calf presented clinical signs and hematological parameters of bovine ehrlichiosis. The in vitro culture of the agent was established in the canine cell line DH82. Ehrlichial morulae were observed using light and electron microscopy within DH82 cells. Total DNA was extracted, and the full genome of the E. minasensis strain Cuiabá was sequenced. A core-genome-based phylogenetic tree of Ehrlichia spp. and Anaplasma spp. confirmed that E. minasensis is a sister taxa of E. canis. A comparison of functional categories among Ehrlichia showed that E. minasensis has significantly less genes in the ‘clustering-based subsystems’ category, which includes functionally coupled genes for which the functional attributes are not well understood. Results strongly suggest that E. minasensis is a novel pathogen infecting cattle. The epidemiology of this Ehrlichia deserves further attention because these bacteria could be an overlooked cause of tick-borne bovine ehrlichiosis, with a wide distribution.

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.

Isolation and Characterization of a Soluble Phosphate Hydrolysing Activity from an in vitro Coffee Cell Line Grown in the Presence of Aluminum

2007 ◽  
Vol 2 (5) ◽  
pp. 302-313
Author(s):  
Minero-Garcia Yereni . ◽  
Villanueva Marco A. . ◽  
Vazquez-Flota Felipe . ◽  
Hernandez-Sotomayor . ◽  
Islas-Flores Ignacio .
Keyword(s):  
Cell Line ◽  
Soluble Phosphate ◽  
Isolation And Characterization

scholarly journals Isolation and Characterization of a Distinct Subpopulation from the WM115 Cell Line That Resembles In Vitro Properties of Melanoma Cancer Stem Cells

2017 ◽  
Vol 22 (5) ◽  
pp. 484-493
Author(s):  
Sonia G. Escobar ◽  
Mark H. Chin ◽  
Mark L. Sandberg ◽  
Han Xu
Keyword(s):  
Drug Resistance ◽  
Cancer Cells ◽  
Cell Line ◽  
Malignant Tumors ◽  
Tumor Cell Line ◽  
Cancer Therapeutics ◽  
Serum Starvation ◽  
Vitro Assay ◽  
Isolation And Characterization

Despite key advances in cancer therapies, malignant tumors, such as melanoma, continue to be one of the leading causes of mortality. Recent debate on whether cancer can originate from a tumor-initiating subpopulation has permeated oncology and stem cell research. It has been well established that primary and immortalized tumor cells consist of heterogeneous cell populations. The profound effect of tumor heterogeneity on tumor growth and drug resistance remains elusive, but it is highly likely that subpopulations of cancer cells have different capabilities of self-renewal and drug resistance. Discrepancies between excellent in vitro potency and efficacy and poor patient response have been observed on multiple cancer therapeutics. Although this observation can be attributed to many factors, a better understanding of the contribution from subpopulations within a cancer will help bridge the gap between in vitro assay results and patient prognosis. To comprehend this impact, it is critical to isolate and characterize cancer subpopulations that possess higher growth and drug resistance properties so that novel therapeutics can be developed to eventually eradicate all cancer cells. In this article, we describe a method to enrich a subpopulation, CB4, from the melanoma cell line WM115. CB4 exhibited higher anchorage-independent growth, higher survival under serum starvation condition, and lower drug sensitivity to commonly used melanoma treatment compared with WM115. Details of functional properties and gene expression of CB4 compared with WM115 are reported. Our study demonstrates that it is feasible to isolate and enrich a subpopulation that exhibits higher growth capacity and treatment resistance from an immortalized tumor cell line.

scholarly journals Biochemical and Haematological Blood Parameters at Different Stages of Lactation in Cows

2017 ◽  
Vol 74 (1) ◽  
pp. 31 ◽  
Author(s):  
Cristian Ovidiu COROIAN ◽  
Vioara MIRESAN ◽  
Aurelia COROIAN ◽  
Camelia RADUCU ◽  
Luisa ANDRONIE ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Biochemical Parameters ◽  
Post Partum ◽  
Hematological Parameters ◽  
Technological Factor ◽  
Clinical Signs ◽  
Blood Parameters ◽  
Holstein Breed ◽  
Lactating Cows ◽  
Blood Biochemical

The health status of cows is evaluated and depending on haematological and biochemical profile of blood. Nutrition is the main technological factor that can produce profound changes in the metabolic profile in animals (Dhiman et al., 1991; Khaled et al., 1999; Ingvartsen, 2006). Blood parameters analyze can lead to identify if there are errors in nutrition of lactating cows (Payne et al., 1970). The aim of this study was the evaluation of metabolic and biochemical changes that occur during colostrum period and in terms of number of lactations in cows. The biological material was represented by a total of 60 heads of dairy cows from a family farm from Sălaj County, Romania. The cows are all from Holstein breed and presented no clinical signs of any specific pathology. Blood samples were collected from the jugular vein of each cow and analyzed. 10 individuals from each of the six lactations have been randomly selected. Haematological and biochemical parameters showed variations depending on factors analyzed here. In lactation 1 Hb was 7.55±3.05 (g/dl), while in lactation 6 the value was 12.5±2.10 (g/dl). RBC ranged as follows: in lactation 1 - 28.50±2.05 and in lactation 6 - 30.02±2.05. Lymphocytes varied within very wide limits under the influence of lactation: in lactation 1 - 2.8±1.56 and in lactation 6 - 7.55±1.80. The number of lactations and lactation rank have influenced blood biochemical and hematological parameters in dairy cows. Biochemical parameters are influenced by post-partum day, showing the lowest values in the early days of colostral period and the highest in the last few days of the same period.

Gene therapy delivering a paraoxonase 1 variant offers long-term prophylactic protection against nerve agents in mice

2020 ◽  
Vol 12 (527) ◽  
pp. eaay0356 ◽  
Author(s):  
Venkaiah Betapudi ◽  
Reena Goswami ◽  
Liliya Silayeva ◽  
Deborah M. Doctor ◽  
Nageswararao Chilukuri
Keyword(s):  
Gene Therapy ◽  
Hematological Parameters ◽  
Clinical Signs ◽  
Nerve Agents ◽  
Medical Intervention ◽  
Chemical Warfare ◽  
Paraoxonase 1 ◽  
Behavioral Abnormalities

Chemical warfare nerve agents are organophosphorus chemical compounds that induce cholinergic crisis, leaving little or no time for medical intervention to prevent death. The current chemical treatment regimen may prevent death but does not prevent postexposure complications such as brain damage and permanent behavioral abnormalities. In the present study, we have demonstrated an adeno-associated virus 8 (AAV8)–mediated paraoxonase 1 variant IF-11 (PON1-IF11) gene therapy that offers asymptomatic prophylactic protection to mice against multiple lethal doses of G-type chemical warfare nerve agents, namely, tabun, sarin, cyclosarin, and soman, for up to 5 months in mice. A single injection of liver-specific adeno-associated viral particles loaded with PON1-IF11 gene resulted in expression and secretion of recombinant PON1-IF11 in milligram quantities, which has the catalytic power to break down G-type chemical warfare nerve agents into biologically inactive products in vitro and in vivo in rodents. Mice containing milligram concentrations of recombinant PON1-IF11 in their blood displayed no clinical signs of toxicity, as judged by their hematological parameters and serum chemistry profiles. Our study unfolds avenues to develop a one-time application of gene therapy to express a near-natural and circulating therapeutic PON1-IF11 protein that can potentially protect humans against G-type chemical warfare nerve agents for several weeks to months.

scholarly journals Tick Cell Culture Analysis of Growth Dynamics and Cellular Tropism of Rickettsia buchneri, an Endosymbiont of the Blacklegged Tick, Ixodes scapularis

Insects ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 968
Author(s):  
Cody J. Thorpe ◽  
Xin-Ru Wang ◽  
Ulrike G. Munderloh ◽  
Timothy J. Kurtti
Keyword(s):  
Fluorescent Protein ◽  
Ixodes Scapularis ◽  
Animal Health ◽  
Light And Electron Microscopy ◽  
Growth Dynamics ◽  
Sensu Stricto ◽  
23S Rrna ◽  
Tissue Tropism ◽  
Blacklegged Tick

The blacklegged tick, Ixodes scapularis, a species of significant importance to human and animal health, harbors an endosymbiont Rickettsia buchneri sensu stricto. The symbiont is largely restricted to the ovaries, but all life stages can harbor various quantities or lack R. buchneri entirely. The endosymbiont is cultivable in cell lines isolated from embryos of Ixodes ticks. Rickettsia buchneri most readily grows and is maintained in the cell line IRE11 from the European tick, Ixodes ricinus. The line was characterized by light and electron microscopy and used to analyze the growth dynamics of wildtype and GFPuv-expressing R. buchneri. qPCR indicated that the genome copy doubling time in IRE11 was >7 days. Measurements of fluorescence using a plate reader indicated that the amount of green fluorescent protein doubled every 11 days. Two 23S rRNA probes were tested via RNA FISH on rickettsiae grown in vitro and adapted to evaluate the tissue tropism of R. buchneri in field-collected female I. scapularis. We observed strong positive signals of R. buchneri in the ovaries and surrounding the nucleus of the developing oocytes. Tissue tropism in I. scapularis and in vitro growth dynamics strengthen the contemporary understanding of R. buchneri as a transovarially transmitted, non-pathogenic endosymbiont.

scholarly journals Preclinical Development of Mahanine-Enriched Fraction from Indian Spice Murraya koenigii for the Management of Cancer: Efficacy, Temperature/pH stability, Pharmacokinetics, Acute and Chronic Toxicity (14-180 Days) Studies

2020 ◽  
Vol 2020 ◽  
pp. 1-18
Author(s):  
Eswara Murali Satyavarapu ◽  
Prasun Kumar Sinha ◽  
Chitra Mandal
Keyword(s):  
Breast Cancer ◽  
Chronic Toxicity ◽  
Hematological Parameters ◽  
Clinical Signs ◽  
Murraya Koenigii ◽  
Preclinical Development ◽  
Dapi Staining ◽  
Wide Range

Murraya koenigii is well documented in the Indian ancient medical text “Charaka Samhita.” The carbazole alkaloid “mahanine” from this plant exhibited anticancer activity against several cancers. Here, we have taken a comprehensive study to standardize the method for the preparation of a mahanine-enriched fraction (MEF) with the highest yield and defined markers. Our optimized method produced MEF having the highest amount of mahanine, a major marker, with excellent in vitro antiproliferative activity against ovarian and breast cancer cells as evidenced by decreased cell viability by MTT assay. Moreover, it exhibited condensed and fragmented nuclei by DAPI staining and increased annexin V-/PI-stained cells after MEF treatment, indicating apoptosis. It also exhibited good efficacy in ovarian and breast cancer syngeneic mice models, with an ED50 of 300 mg/kg body weight (BW). MEF is stable up to 40°C for ≥3 months. Its biological activity remains unchanged at a wide range of pH (1-10) for up to ~3 hours, indicating a safe oral route of administration. Additionally, the comparative pharmacokinetics of MEF and mahanine in rats showed a 31% higher bioavailability of mahanine in MEF-fed rats compared to rats fed with mahanine alone. Furthermore, mice fed with MEF at 5000 mg/kg BW single dose, 300-1500 mg/kg BW/day for 14 days, and 300 mg/kg BW/day for 28, 90, and 180 days for subacute, subchronic, chronic studies, respectively, did not show any significant clinical signs of toxicity, behavioral changes, mortality, organ weights, serum biochemistry, and hematological parameters indicating no/minimum toxicity for up to 180 days. To the best of our knowledge, this is the first report showing the pH/temperature stability and chronic toxicity studies of MEF along with in vivo efficacy against breast cancer. Taken together, our study will enhance the commercial value of this highly potential medicinal plant and will be helpful as a reference material for its clinical development.

scholarly journals Production of Cloned Lambs from an Established Embryonic Cell Line: A Comparison between In Vivo- and In Vitro-Matured Cytoplasts1

1997 ◽  
Vol 57 (2) ◽  
pp. 385-393 ◽  
Author(s):  
David N. Wells ◽  
Pavia M. Misica ◽  
(Tony) A.M. Day ◽  
H. Robin Tervit
Keyword(s):  
Cell Line ◽  
Embryonic Cell ◽  
Embryonic Cell Line
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