Ex Vivo Phenotypic Screening of Two Small Repurposing Drug Collections Identifies Nifuratel as a Potential New Treatment against Visceral and Cutaneous Leishmaniasis

2021 ◽  
Author(s):  
Bárbara Domínguez-Asenjo ◽  
Camino Gutiérrez-Corbo ◽  
María Álvarez-Bardón ◽  
Yolanda Pérez-Pertejo ◽  
Rafael Balaña-Fouce ◽  
...  
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Ex Vivo ◽  
Phenotypic Screening ◽  
New Treatment

scholarly journals Flow Cytometric Determination of Cellular Sources and Frequencies of Key Cytokine-Producing Lymphocytes Directed against Recombinant LACK and Soluble Leishmania Antigen in Human Cutaneous Leishmaniasis

2001 ◽  
Vol 69 (5) ◽  
pp. 3232-3239 ◽  
Author(s):  
R. L. A. Bottrel ◽  
W. O. Dutra ◽  
F. A. Martins ◽  
B. Gontijo ◽  
E. Carvalho ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cutaneous Leishmaniasis ◽  
Ex Vivo ◽  
Protozoan Parasite ◽  
Recombinant Antigen ◽  
Interleukin 4 ◽  
Leishmania Braziliensis ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Ifn Γ

ABSTRACT Leishmaniasis, caused by infection with the protozoan parasiteLeishmania, affects millions of individuals worldwide, causing serious morbidity and mortality. This study directly determined the frequency of cells producing key immunoregulatory cytokines in response to the recombinant antigen Leishmania homolog of receptors for activated kinase C (LACK) and soluble leishmania antigen (SLA), and it determined relative contributions of these antigens to the overall cytokine profile in individuals infected for the first time with Leishmania braziliensis. All individuals presented with the cutaneous clinical form of leishmaniasis and were analyzed for proliferative responses to LACK antigen and SLA, frequency of lymphocyte subpopulations (analyzed ex vivo), and antigen-induced (LACK and SLA) cytokine production at the single-cell level (determined by flow cytometry). The following were determined. (i) The Th1-type response previously seen in patients with cutaneous leishmaniasis is due to gamma interferon (IFN-γ) production by several different sources, listed in order of contribution: CD4+ T lymphocytes, CD4−, CD8− lymphocytes, and CD8+ T lymphocytes. (ii) SLA induced a higher frequency of lymphocytes producing IFN-γ and tumor necrosis factor alpha (TNF-α) than did LACK. (iii) LACK induced an activation of monocyte populations as reflected by an increased percentage of CD14-positive cells. (iv) Neither SLA nor LACK induced detectable frequencies of cells producing interleukin-4 (IL-4) or IL-5. These data demonstrated a multifaceted immune response to SLA in human leishmaniasis involving Th1 CD4+ T lymphocytes (IFN-γ+ and IL-10−/IL-4−), Tc1 CD8+ T cells (IFN-γ+, and IL-10−/IL-4−), and a high frequency of TNF-α-producing lymphocytes. Moreover, it was determined that the recombinant antigen LACK acts as a weak inducer of Th1-type lymphocyte responses compared to SLA.

scholarly journals Microwave thermotherapy: New treatment for cutaneous leishmaniasis

2015 ◽  
Vol 6 (2) ◽  
pp. 125-129 ◽  
Author(s):  
Khalifa E. Sharquie ◽  
Sabeeh A. Al-Mashhadani ◽  
Adil A. Noaimi ◽  
Wasan B. Al-Zoubaidi
Keyword(s):  
Cutaneous Leishmaniasis ◽  
New Treatment

scholarly journals Endogenous 24S-hydroxycholesterol modulates NMDAR-mediated function in hippocampal slices

2016 ◽  
Vol 115 (3) ◽  
pp. 1263-1272 ◽  
Author(s):  
Min-Yu Sun ◽  
Yukitoshi Izumi ◽  
Ann Benz ◽  
Charles F. Zorumski ◽  
Steven Mennerick
Keyword(s):  
Brain Function ◽  
Ex Vivo ◽  
Hippocampal Slices ◽  
Treatment Strategies ◽  
Glucose Deprivation ◽  
Wild Type ◽  
Drug Candidates ◽  
Excitatory Transmission ◽  
The Central Nervous System ◽  
New Treatment

N-methyl-d-aspartate receptors (NMDARs), a major subtype of glutamate receptors mediating excitatory transmission throughout the central nervous system (CNS), play critical roles in governing brain function and cognition. Because NMDAR dysfunction contributes to the etiology of neurological and psychiatric disorders including stroke and schizophrenia, NMDAR modulators are potential drug candidates. Our group recently demonstrated that the major brain cholesterol metabolite, 24 S-hydroxycholesterol (24S-HC), positively modulates NMDARs when exogenously administered. Here, we studied whether endogenous 24S-HC regulates NMDAR activity in hippocampal slices. In CYP46A1−/−(knockout; KO) slices where endogenous 24S-HC is greatly reduced, NMDAR tone, measured as NMDAR-to-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) excitatory postsynaptic current (EPSC) ratio, was reduced. This difference translated into more NMDAR-driven spiking in wild-type (WT) slices compared with KO slices. Application of SGE-301, a 24S-HC analog, had comparable potentiating effects on NMDAR EPSCs in both WT and KO slices, suggesting that endogenous 24S-HC does not saturate its NMDAR modulatory site in ex vivo slices. KO slices did not differ from WT slices in either spontaneous neurotransmission or in neuronal intrinsic excitability, and exhibited LTP indistinguishable from WT slices. However, KO slices exhibited higher resistance to persistent NMDAR-dependent depression of synaptic transmission induced by oxygen-glucose deprivation (OGD), an effect restored by SGE-301. Together, our results suggest that loss of positive NMDAR tone does not elicit compensatory changes in excitability or transmission, but it protects transmission against NMDAR-mediated dysfunction. We expect that manipulating this endogenous NMDAR modulator may offer new treatment strategies for neuropsychiatric dysfunction.

scholarly journals Cannabinoid Receptor Agonist Inhibits Atrial Electrical Remodeling in a Tachypaced Ex Vivo Rat Model

2021 ◽  
Vol 12 ◽  
Author(s):  
Danielle I. Lee ◽  
Michael Murninkas ◽  
Sigal Elyagon ◽  
Yoram Etzion ◽  
Hope D. Anderson
Keyword(s):  
Cannabinoid Receptor ◽  
Ex Vivo ◽  
Clinical Value ◽  
Rate Dependent ◽  
Rat Hearts ◽  
Before And After ◽  
New Treatment ◽  
Set Up ◽  
The Right ◽  
Peripheral Activation

Introduction: Atrial fibrillation (AF) leads to rate-dependent atrial changes collectively defined as atrial remodelling (AR). Shortening of the atrial effective refractory period (AERP) and decreased conduction velocity are among the hallmarks of AR. Pharmacological strategies to inhibit AR, thereby reducing the self-perpetual nature of AF, are of great clinical value. Cannabinoid receptor (CBR) ligands may exert cardioprotective effects; CB13, a dual CBR agonist with limited brain penetration, protects cardiomyocytes from mitochondrial dysfunction induced by endothelin-1. Here, we examined the effects of CB13 on normal physiology of the rat heart and development of tachypacing-induced AR.Methods: Rat hearts were perfused in a Langendorff set-up with CB13 (1 µM) or vehicle. Hemodynamic properties of non-paced hearts were examined conventionally. In a different set of hearts, programmed stimulation protocol was performed before and after atrial tachypacing for 90 min using a mini-hook platinum quadrupole electrode inserted on the right atrium. Atrial samples were further assessed by western blot analysis.Results: CB13 had no effects on basal hemodynamic properties. However, the compound inhibited tachypacing-induced shortening of the AERP. Protein expression of PGC1α was significantly increased by CB13 compared to vehicle in paced and non-paced hearts. Phosphorylation of AMPKα at residue threonine 172 was increased suggesting upregulation of mitochondrial biogenesis. Connexin43 was downregulated by tachypacing. This effect was diminished in the presence of CB13.Conclusion: Our findings support the notion that peripheral activation of CBR may be a new treatment strategy to prevent AR in patients suffering from AF, and therefore warrants further study.

scholarly journals CD16+ monocytes in human cutaneous leishmaniasis: increased ex vivo levels and correlation with clinical data

2005 ◽  
Vol 79 (1) ◽  
pp. 36-39 ◽  
Author(s):  
George Soares ◽  
Aldina Barral ◽  
Jackson M. Costa ◽  
Manoel Barral-Netto ◽  
Johan Van Weyenbergh
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Clinical Data ◽  
Ex Vivo ◽  
Human Cutaneous Leishmaniasis

scholarly journals Personalizing Oncolytic Virotherapy for Glioblastoma: In Search of Biomarkers for Response

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 614 ◽  
Author(s):  
Eftychia Stavrakaki ◽  
Clemens M. F. Dirven ◽  
Martine L. M. Lamfers
Keyword(s):  
Oncolytic Virus ◽  
Ex Vivo ◽  
Oncolytic Viruses ◽  
Specific Patient ◽  
Specific Tumor ◽  
Clinical Responses ◽  
Large Heterogeneity ◽  
New Treatment ◽  
Selection Of

Oncolytic virus (OV) treatment may offer a new treatment option for the aggressive brain tumor glioblastoma. Clinical trials testing oncolytic viruses in this patient group have shown promising results, with patients achieving impressive long-term clinical responses. However, the number of responders to each OV remains low. This is thought to arise from the large heterogeneity of these tumors, both in terms of molecular make-up and their immune-suppressive microenvironment, leading to variability in responses. An approach that may improve response rates is the personalized utilization of oncolytic viruses against Glioblastoma (GBM), based on specific tumor- or patient-related characteristics. In this review, we discuss potential biomarkers for response to different OVs as well as emerging ex vivo assays that in the future may enable selection of optimal OV for a specific patient and design of stratified clinical OV trials for GBM.

scholarly journals A human ex vivo coculture model to investigate peritoneal metastasis and innovative treatment options

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Dina Mönch ◽  
Jana Koch ◽  
Annika Maaß ◽  
Nicole Janssen ◽  
Thomas Mürdter ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Cell Lines ◽  
Peritoneal Metastasis ◽  
Ex Vivo ◽  
Treatment Options ◽  
Treatment Strategies ◽  
Average Survival ◽  
Extracellular Matrix Components ◽  
Coculture Model ◽  
New Treatment

Abstract Objectives Peritoneal metastasis (PM) is commonly observed in patients with colorectal cancer (CRC). The outcome of these patients is poor, with an average survival of only six months without therapy, which requires a better understanding of PM biology and new treatment strategies. Methods We established and characterized a human ex vivo peritoneal model to investigate the mechanisms of peritoneal seeding and possible treatment options. For this, CRC cell lines and patient-derived tumor organoids were cultured together with human peritoneum to investigate the invasion of malignant cells and the effects of local chemotherapy. Results Fresh human peritoneum was cultured for up to three weeks in a stainless steel ring system, allowing for survival of all peritoneal structures. Peritoneal cell survival was documented by light microscopy and immunohistochemical staining. Further, immunohistological characterization of the tissue revealed CD3-positive T-lymphocytes and vimentin-positive fibroblasts within the peritoneum. In addition, extracellular matrix components (collagens, matrix metalloproteinases) were localized within the tissue. Coculture with CRC cell lines and patient-derived CRC organoids revealed that cancer cells grew on the peritoneum and migrated into the tissue. Coculture with CRC cells confirmed that hyperthermal treatment at 41 °C for 90 min significantly enhanced the intracellular entry of doxorubicin. Moreover, treatment with mitomycin C under hyperthermic conditions significantly reduced the amount of cancer cells within the peritoneum. Conclusions This human ex vivo peritoneal model provides a stringent and clinically relevant platform for the investigation of PM and for further elucidation of possible treatment options.

scholarly journals Targeted imaging of urothelium carcinoma in human bladders by an ICG pHLIP peptide ex vivo

2016 ◽  
Vol 113 (42) ◽  
pp. 11829-11834 ◽  
Author(s):  
Jovana Golijanin ◽  
Ali Amin ◽  
Anna Moshnikova ◽  
Joseph M. Brito ◽  
Timothy Y. Tran ◽  
...  
Keyword(s):  
White Light ◽  
Near Infrared ◽  
Ex Vivo ◽  
Bladder Preservation ◽  
Urothelial Carcinomas ◽  
Malignant Lesions ◽  
New Treatment ◽  
Muscle Invasive ◽  
Treatment Alternatives

Bladder cancer is the fifth most common in incidence and one of the most expensive cancers to treat. Early detection greatly improves the chances of survival and bladder preservation. The pH low insertion peptide (pHLIP) conjugated with a near-infrared fluorescent dye [indocyanine green (ICG)] targets low extracellular pH, allowing visualization of malignant lesions in human bladder carcinoma ex vivo. Cystectomy specimens obtained after radical surgery were immediately irrigated with nonbuffered saline and instilled with a solution of the ICG pHLIP construct, incubated, and rinsed. Bladders were subsequently opened and imaged, the fluorescent spots were marked, and a standard pathological analysis was carried out to establish the correlation between ICG pHLIP imaging and white light pathological assessment. Accurate targeting of bladder lesions was achieved with a sensitivity of 97%. Specificity is 100%, but reduced to 80% if targeting of necrotic tissue from previous transurethral resections or chemotherapy are considered as false positives. The ICG pHLIP imaging agent marked high-grade urothelial carcinomas, both muscle invasive and nonmuscle invasive. Carcinoma in situ was accurately diagnosed in 11 cases, whereas only four cases were seen using white light, so imaging with the ICG pHLIP peptide offers improved early diagnosis of bladder cancers and may also enable new treatment alternatives.

scholarly journals Inhaled sphingosine has no adverse side effects in isolated ventilated and perfused pig lungs

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Henning Carstens ◽  
Katharina Kalka ◽  
Rabea Verhaegh ◽  
Fabian Schumacher ◽  
Matthias Soddemann ◽  
...  
Keyword(s):  
Side Effects ◽  
Lung Transplantation ◽  
Performance Test ◽  
Ex Vivo ◽  
High Dose ◽  
Tissue Samples ◽  
Bronchial Epithelial ◽  
Luminal Membrane ◽  
High Doses ◽  
New Treatment

AbstractEx-vivo lung perfusion (EVLP) systems like XVIVO are more and more common in the setting of lung transplantation, since marginal donor-lungs can easily be subjected to a performance test or be treated with corticosteroids or antibiotics in high dose regimes. Donor lungs are frequently positive in bronchoalveolar lavage (BAL) bacterial cultures (46–89%) which leads to a donor-to-recipient transmission and after a higher risk of lung infection with reduced posttransplant outcome. We have previously shown that sphingosine very efficiently kills a variety of pathogens, including Pseudomonas aeruginosa, Staphylococcus aureus and epidermidis, Escherichia coli or Haemophilus influenzae. Thus, sphingosine could be a new treatment option with broadspectrum antiinfective potential, which may improve outcome after lung transplantation when administered prior to lung re-implantation. Here, we tested whether sphingosine has any adverse effects in the respiratory tract when applied into isolated ventilated and perfused lungs. A 4-h EVLP run using minipig lungs was performed. Functional parameters as well as perfusate measurements where obtained. Biopsies were obtained 30 min and 150 min after inhalation of sphingosine. Tissue samples were fixed in paraformaldehyde, embedded in paraffin and sectioned. Hemalaun, TUNEL as well as stainings with Cy3-coupled anti-sphingosine or anti-ceramide antibodies were implemented. We demonstrate that tube-inhalation of sphingosine into ex-vivo perfused and ventilated minipig lungs results in increased levels of sphingosine in the luminal membrane of bronchi and the trachea without morphological side effects up to very high doses of sphingosine. Sphingosine also did not affect functional lung performance. In summary, the inhalation of sphingosine results in an increase of sphingosine concentrations in the luminal plasma membrane of tracheal and bronchial epithelial cells. The inhalation has no local side effects in ex-vivo perfused and ventilated minipig lungs.

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