Carolina Moracho-Vilrriales’s research while affiliated with Presbyterian College and other places

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Publications (4)


Table 1 Demographic characteristics of the patients enrolled in the study
Cytokine concentration in nonstimulated primary human macrophages.
Table 2 Levels of anti-inflammatory cytokines in LPS-stimulated primary human macrophages
Cytokine concentration in LPS-stimulated primary human macrophages.
Table 3 Levels of proinflammatory cytokines in LPS-stimulated primary human macrophages

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Cytokine production capabilities of human primary monocyte-derived macrophages from patients with diabetes mellitus type 2 with and without diabetic peripheral neuropathy
  • Article
  • Full-text available

December 2018

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79 Reads

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20 Citations

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Rachel L Grosick

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Carolina Moracho-Vilrriales

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[...]

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Edgar Alfonso Romero-Sandoval

Introduction Monocytes from patients with diabetes mellitus type 2 (DM2) are dysfunctional, persistently primed, and prone to a proinflammatory phenotype. This may alter the phenotype of their differentiation to macrophages and result in diabetic peripheral neuropathy (DPN), nerve damage, nerve sensitization, and chronic pain. We have previously demonstrated that CD163 is a molecule that promotes an anti-inflammatory cellular phenotype in human primary macrophages, but this has not been proven in macrophages from patients with DM2 or DPN. Thus, we hypothesize that macrophages from patients with DM2 or DPN display an altered proinflammatory functional phenotype related to cytokine production and that the induction of CD163 expression will promote a more homeostatic phenotype by reducing their proinflammatory responsiveness. Patients and methods We tested these hypotheses in vitro using blood monocyte-derived macrophages from healthy subjects and patients with DM2 with and without DPN. Cells were incubated in the presence or the absence of 5 µg/mL of lipopolysaccharide (LPS). The concentrations of interleukin-10, interleukin-6, tumor necrosis factor-alpha (TNF-α), TGF-β, and monocyte chemoattractant protein-1 (MCP-1) were measured using ELISA assays. Macrophages were transfected with an empty vector plasmid or a plasmid containing the CD163 gene using mannosylated polyethylenimine nanoparticles. Results Our results show that nonstimulated DM2 or DPN macrophages have a constitutive primed proinflammatory state and display a deficient production of proinflammatory cytokines upon a proinflammatory challenge when compared to healthy macrophages. CD163 induction produced an anti-inflammatory phenotype in the healthy control group, and this effect was partial in DM2 or DPN macrophages. Conclusion Our results suggest that diabetic macrophages adopt a complex phenotype that is only partially reversed by CD163 induction. Future experiments are focused on elucidating this differential responsiveness between healthy and diabetic macrophages.

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Laboratory analysis of soil respiration using oxygen-sensitive microplates

November 2017

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46 Reads

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3 Citations

Geoderma

Soil respiration is usually monitored by measuring CO2 efflux. Most techniques available for such kind of analyses are inconvenient and difficult to adapt to micro-method format. The present study evaluates a new microplate-based method for studying soil respiration in the laboratory. Second-generation oxygen-sensitive microplates (OSM - containing a fluorescent probe attached to the bottom of the well which provides time-resolved fluorescence data) were used to measure soil respiration either in microcosm assays or in soil water extracts. The latter procedure (water extracts) was the least cumbersome, hence it was selected for further experiments. Soil respiration curves generally showed no lag phase, starting with an exponential oxygen consumption phase, followed by a period where respiration became stable after 8–10 h of incubation at 25 °C. Once the procedure for measurement of oxygen consumption in soil was established, the acute toxic effect of diverse chemicals on soil was analysed with OSM. Streptomycin and penicillin failed to reduce soil respiration. Kanamycin plus neomycin, trimethoprim and 5-fluorocytosine exhibited limited inhibitory effects. In contrast, some chemicals (copper sulphate and amphotericin B) and fungicides (such as dodine and fosetyl) noticeably reduced fluorescence readings, showing concentrations to give half-maximal inhibition of respiration (ICR50) ranging from 0.0064 to 0.082 g/L. Finally, some insecticides and soil amendments assayed were either neutral or increased respiration. It is concluded that OSM are reliable, convenient, and yield quantitative results. Moreover, the system is relatively inexpensive and amenable to automation. However, results obtained using soil water extracts may be different from those derived from undisturbed soil aggregates, clods or slurries studied under field conditions.


Identification of DPN and p-DN in PCSP Wellness Center diabetic patients.
Diagram of combination of medications used by patient.
Comparison of doses between patients receiving AAN-recommended doses of gabapentin (optimal) and doses below AAN recommendation (suboptimal).
Proportion of p-DN patients receiving drug combinations with optimal, suboptimal, or partially optimal (mixed) doses.
Table 6 Number of pain medication use in patients with p-DN (n=26)
Identification, prevalence, and treatment of painful diabetic neuropathy in patients from a rural area in South Carolina

April 2017

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52 Reads

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18 Citations

Diabetic peripheral neuropathy (DPN) represents significant burdens to many patients and the public health-care system. Patients with diabetes in rural areas have higher risk of developing complications and having less access to proper treatment. We studied a rural population of patients with diabetes who attended a pharmacist-led free clinic for a diabetic education program. Our objectives were to 1) determine the prevalence of DPN and painful diabetic neuropathy (p-DN) in patients with type 2 diabetes; 2) assess the proportion of patients with DPN and p-DN left undocumented upon physician referral to a pharmacist-led free clinic; and 3) determine the appropriateness of pain medication regimen. We performed a retrospective analysis of clinical records of patients from the Presbyterian College School of Pharmacy (PCSP) Wellness Center located in Clinton, SC. Diagnoses of DPN and/or p-DN were obtained from referral notes in the clinical records and compared with results from foot examinations performed in the free clinic and clinical features. Medication regimens were also obtained and compared using American Academy of Neurology (AAN) treatment guidelines. Within our study population (n=111), the prevalence of DPN was 62.2% (national average of 28%–45%) and that of p-DN was 23.4% (national average of 11%–24%). In p-DN patients (n=26), 53.8% (n=14) had a documented diagnosis of p-DN by the referring physician, and 46.2% (n=12) were identified by the pharmacists. A total of 95% (19 of 20) of the patients treated for p-DN received adequate pharmacological agents, though suboptimal as per clinical guidelines. More than 50% of the patients used subtherapeutic doses of their medications. Gabapentin was the most frequently used medication in our population (65.4%). Patients in rural South Carolina had a higher prevalence of DPN and p-DN with >60% undocumented cases of p-DN. More than 95% of treated patients did not receive optimum therapy according to AAN guidelines.


Effects of JWH015 in cytokine secretion in primary human keratinocytes and fibroblasts and its suitability for topical/transdermal delivery

January 2017

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3,389 Reads

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30 Citations

Molecular Pain

Background JWH015 is a cannabinoid (CB) receptor type 2 agonist that produces immunomodulatory effects. Since skin cells play a key role in inflammatory conditions and tissue repair, we investigated the ability of JWH015 to promote an anti-inflammatory and pro-wound healing phenotype in human primary skin cells. Methods Human primary keratinocytes and fibroblasts were stimulated with lipopolysaccharide. The mRNA expression of cannabinoid receptors was determined using RT-PCR. The effects of JWH015 (0.05, 0.1, 0.5, and 1 µM) in pro- and anti-inflammatory factors were tested in lipopolysaccharide-stimulated cells. A scratch assay, using a co-culture of keratinocytes and fibroblasts, was used to test the effects of JWH015 in wound healing. In addition, the topical and transdermal penetration of JWH015 was studied in Franz diffusion cells using porcine skin and LC-MS. Results The expression of CB1 and CB2 receptors (mRNA) and the production of pro- and anti-inflammatory factors enhanced in keratinocytes and fibroblasts following lipopolysaccharide stimulation. JWH015 reduced the concentration of major pro-inflammatory factors (IL-6 and MCP-1) and increased the concentration of a major anti-inflammatory factor (TGF-β) in lipopolysaccharide-stimulated cells. JWH015 induced a faster scratch gap closure. These JWH015’seffects were mainly modulated through both CB1 and CB2 receptors. Topically administered JWH015 was mostly retained in the skin and displayed a sustained and low level of transdermal permeation. Conclusions Our findings suggest that targeting keratinocytes and fibroblasts with cannabinoid drugs could represent a therapeutic strategy to resolve peripheral inflammation and promote tissue repair.

Citations (4)


... Macrophage activation determines the initiation of signaling pathways with the release of TNF-α, IL-1β, IL-6, or other inflammatory signaling factors such as NF-κB, resulting in organ damage [18] (Figure 4). DM is associated with persistent inflammation accompanied by a pro-inflammatory phenotype at the level of monocytes/macrophages [65]. This systemic inflammatory response may result from a dysfunctional pro-inflammatory M1 macrophage phenotype, contributing to the etiopathogenesis of diabetic neuropathy, especially its painful forms. ...

Reference:

International Journal of Molecular Sciences Update on Biomarkers of Chronic Inflammatory Processes Underlying Diabetic Neuropathy
Cytokine production capabilities of human primary monocyte-derived macrophages from patients with diabetes mellitus type 2 with and without diabetic peripheral neuropathy

... USA (62.2%; national average is 28-45%) also report a higher prevalence for DPN. 30,31 In these developed countries foot ware is worn at all times in contrast to the Asian populations where foot ware is not worn indoors. 32,33 A study conducted in India shows the prevalence of DPN as 45.4%, in concordance with the findings of the current study (with two tests being positive). ...

Identification, prevalence, and treatment of painful diabetic neuropathy in patients from a rural area in South Carolina

... database; these studies have focused on new therapeutic protocols with existing drugs or the use of active substances with known antifibrotic and/or antiinflammatory effects, such as cannabinoids (CBs), for treatment of IPF (Heukels et al., 2019;Liu et al., 2022). CB1 (CB1R) (Gkoumassi et al., 2007;Servettaz et al., 2010;Karmaus et al., 2013;Liu et al., 2014a;Bronova et al., 2015;Staiano et al., 2016;Cinar et al., 2017;Muthumalage and Rahman, 2019;Mohammed et al., 2020;Zawatsky et al., 2020;Chen et al., 2022) and CB2 (CB2R) (Gkoumassi et al., 2007;Servettaz et al., 2010;Costola-de-Souza et al., 2013;Karmaus et al., 2013;Staiano et al., 2016;Bort et al., 2017;Liu and Shi, 2019;Muthumalage and Rahman, 2019;Liu et al., 2020;Parlar et al., 2021;Liu et al., 2022) receptor agonists exhibit inflammation-modulating (Gkoumassi et al., 2007;Costola-de-Souza et al., 2013;Karmaus et al., 2013;Liu et al., 2014b;Staiano et al., 2016;Bort et al., 2017;Cinar et al., 2017;Liu and Shi, 2019;Muthumalage and Rahman, 2019;Liu et al., 2020;Mohammed et al., 2020;Zawatsky et al., 2020;Parlar et al., 2021;Liu et al., 2022) and antifibrotic effects (Servettaz et al., 2010;Bronova et al., 2015;Liu and Shi, 2019;Chen et al., 2022) in lung tissue or bronchoalveolar lavage fluid (BALF) by focusing on inflammation markers, including transforming growth factor-β1 (TGF-β1), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) and collagen type I (ColI) in human bronchial epithelial cells (Gkoumassi et al., 2007;Muthumalage and Rahman, 2019), human embryonic lung fibroblasts (Liu and Shi, 2019;Muthumalage and Rahman, 2019), and monocyte and lung resident macrophages (Staiano et al., 2016;Muthumalage and Rahman, 2019). In addition, human bronchial epithelial cells (Gkoumassi et al., 2007;Boyacioglu et al., 2021), lung macrophages (Staiano et al., 2016), and mice lung tissue (Tahamtan et al., 2018;Huang et al., 2020) express CB receptors (CBRs). ...

Effects of JWH015 in cytokine secretion in primary human keratinocytes and fibroblasts and its suitability for topical/transdermal delivery

Molecular Pain