Maxy Santos - Academia.edu (original) (raw)

Papers by Maxy Santos

Research paper thumbnail of Sensitivity and specificity of the real-time PCR assay compared to kDNA PCR stratified by site, type of sample and type of lesion

(*)<p>Statistically significant (<i>p</i><0.001) by Chi-square test.</p

Research paper thumbnail of Number of ear, tail, bone marrow, and liver tissues tested for <i>Leishmania</i> spp. via polymerase chain reaction shown by study site and species captured

1<p>Where n<sub>EC</sub> and n<sub>MZ</sub> are the total captures ... more 1<p>Where n<sub>EC</sub> and n<sub>MZ</sub> are the total captures at El Carmen and Mazuko, respectively</p>2<p><i>Proechimys spp</i>. included 1 <i>P. simonsi</i> at Mazuko and 6 <i>P. simonsi</i> at El Carmen. The remaining 11 at El Carmen were identified only to genus level.</p

Research paper thumbnail of <i>Leishmania</i> species identified by real-time PCR from individuals with suspected leishmaniasis from IMTAvH and HMC samples

(*)<p>No amplification for the <i>MPI</i> real-time PCR assay. <i>6PGD&lt... more (*)<p>No amplification for the <i>MPI</i> real-time PCR assay. <i>6PGD</i> real-time PCR results suggested infection by either <i>L.</i> (<i>V.</i>) <i>peruviana</i> or <i>L.</i> (<i>V.</i>) <i>braziliensis</i> but in the absence of MPI results, we could not discriminate between these two <i>Leishmania</i> species.</p

Research paper thumbnail of Upper bound Clopper-Pearson one-sided 95% confidence intervals for an observed <i>Leishmania</i> prevalence of 0%, evaluated by two strata: rodent species (for those species with ≥15 individuals represented) and study site

Research paper thumbnail of Genetic diversity and population structure of Leishmania (Viannia) braziliensis in the Peruvian jungle

PLOS Neglected Tropical Diseases

Background Human cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis is highly pr... more Background Human cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis is highly prevalent in the Peruvian jungle, where it affects military forces deployed to fight against drug trafficking and civilian people that migrate from the highland to the lowland jungle for economic activities such as mining, agriculture, construction, and chestnut harvest. We explored the genetic diversity and population structure of 124 L. (V.) braziliensis isolates collected from the highland (Junín, Cusco, and Ayacucho) and lowland Peruvian jungle (Loreto, Ucayali, and Madre de Dios). All samples were genotyped using Multilocus Microsatellite Typing (MLMT) of ten highly polymorphic markers. Principal findings High polymorphism and genetic diversity were found in Peruvian isolates of L. (V.) braziliensis. Most markers are not in Hardy-Weinberg equilibrium; this deviation is most likely caused by local inbreeding, as shown by the positive FIS values. Linkage Disequilibrium in subpopulations...

Research paper thumbnail of Diagnóstico molecular para Leishmaniasis

Revista Peruana de Medicina Experimental y Salud Pública, 1997

El potencial diagnóstico de epitopes inmunodominantes seleccionados fue ensayado satisfactoriamen... more El potencial diagnóstico de epitopes inmunodominantes seleccionados fue ensayado satisfactoriamente a fin de obtener una prueba serodiagnóstica alternativa para la Leishmaniasis Tegumentaria Americana. Dos proteínas recombinantes prometedoras de L. (v.) peruviana referidas como T-26-U2/T26-U4 fueron reconocidas por sueros individuales de pacientes con Leishmaniasis Tegumentaria Americana usando Western Blot. La sensibilidad de la prueba fue de 86% con sueros permanentes con Leishmaniasis peruana.

Research paper thumbnail of Running Title: Effects of HDACi and RA on SH-SY5Y cells

Combined effects of retinoic acid and histone deacetylase inhibitors on human neuroblastoma SH-SY... more Combined effects of retinoic acid and histone deacetylase inhibitors on human neuroblastoma SH-SY5Y cells

Research paper thumbnail of Inhibitors on Human Neuroblastoma

All-trans retinoic acid (RA) causes differentiation of neuroblastoma cells, and retinoids have be... more All-trans retinoic acid (RA) causes differentiation of neuroblastoma cells, and retinoids have been used in clinical trials in children with advanced neuroblastoma. Combination of RA with histone deacetylase inhibitors (HDACi) could result in improved antitumorigenic activity. We have examined the effect of the HDACi trichostatin A (TSA), sodium butyrate, and suberoylanilide hydroxamic acid (SAHA), alone and in combination with RA in human neuroblastoma SH-SY5Y cells. At concentrations that cause sustained increase of histone H3 acetylation, HDACi produced extensive apoptotic cell death as shown by flow cytometry analysis and induction of poly(ADP-ribose) polymerase proteolysis. HDACi inhibited SH-SY5Y cell growth at a much larger extent than RA. This compound did not cause apoptosis and did not further increase HDACimediated cell death. In contrast, both types of drugs cooperated to inhibit cell growth, although synergistic effects were not found. In surviving cells, HDACi repressed cyclin D1 expression and increased the cyclin kinase inhibitors (CKI) p21 Waf1/Cip1 and p27 Kip1. Cyclin D1 was not affected by RA, but this retinoid also increased CKI levels. Induction of p21 Waf1/Cip1 and p27 Kip1 by HDACi was further enhanced in the presence of RA. This effect seems to be at least partially due to transcriptional stimulation of CKI gene expression because both types of drugs cooperated to increase CKI mRNA levels and to activate the CKI promoters in transient transfection assays. These results show the strong antitumorigenic effects of HDACi in neuroblastoma cells and reinforce the idea that combination therapy could be useful to inhibit tumor growth.

Research paper thumbnail of Diagnóstico molecular para Leishmaniasis

Research paper thumbnail of Diagnostic Efficacy of Recombinase-Polymerase-Amplification Coupled with Lateral Flow Strip Reading in Patients with Cutaneous Leishmaniasis from the Amazonas Rainforest of Perú

Vector-Borne and Zoonotic Diseases, 2021

Cutaneous leishmaniasis (CL) is highly prevalent in rural and sylvatic regions of Latin America, ... more Cutaneous leishmaniasis (CL) is highly prevalent in rural and sylvatic regions of Latin America, with an estimated 55,000 annual cases. Diagnosis in resource-limited areas still relies on microscopy of dermal scrapings, while more sensitive methods like PCR are not attainable due to costs and lack of adequate health infrastructure. Isothermal amplification of Leishmania DNA can be performed without sophisticated equipment and training and may become a point of care (POC) test for health care centers with scarce resources. We evaluated the efficacy of recombinase-polymerase-amplification (RPA-LF) to diagnose CL in 226 patients attending a clinic in Puerto Maldonado within the Peruvian Amazon basin. Conventional PCR targeting kinetoplast DNA (kDNA-PCR) was used as the gold standard. Eight of 226 patients were considered true negatives (microscopy, kDNA-PCR, and RPA-LF negative), while RPA-LF resulted positive in 186 of 204 kDNA-PCR positive patients, yielding 91.2% (confidence interval [CI] = 86.5-94.4%) sensitivity and 93% (CI 88.6-95.8%) positive predictive value. There were 14% (32/226) discrepant samples alternating positive and negative results in similar proportions between both tests. Quantitative PCR used to resolve the discrepancies suggested that they occurred in samples with scarce parasite numbers as determined by high cycle threshold (Ct) values (≥32; cutoff 35.5). Microscopy had the lowest sensitivity of all methods (45.4%). Nested real-time PCR performed in 71 samples determined that Leishmania (Viannia) braziliensis was highly prevalent (69/71), and Leishmania (Viannia) lainsoni was present in only two isolates. Results indicated that RPA-LF has POC potential for CL endemic areas, yet further simplification and optimization coupled with field validation will be necessary to confirm its broad applicability.

Research paper thumbnail of Distribution and identification of sand flies naturally infected with Leishmania from the Southeastern Peruvian Amazon

PLoS neglected tropical diseases, 2017

Cutaneous leishmaniasis (CL) is an important health problem in the New World affecting civilian a... more Cutaneous leishmaniasis (CL) is an important health problem in the New World affecting civilian and military populations that are frequently exposed in endemic settings. The Peruvian region of Madre de Dios located near the border with Brazil is one of the most endemic CL regions in South America with more than 4,451 reported cases between 2010 and 2015 according to the Peruvian epidemiology directorate. However, little is known regarding the diversity and distribution of sand fly vectors in this region. In this study, we aimed to characterize the sand fly fauna in this endemic setting and identify sand fly species naturally infected with Leishmania possibly involved in pathogen transmission. Sand fly collections were carried out during 2014 and 2015 in the communities of Flor de Acre, Villa Primavera, Mavila and Arca Pacahuara using CDC light traps and Shannon traps. Collected specimens were identified and non-blood-fed females were selected for Leishmania infection screening using...

Research paper thumbnail of Histone Deacetylase Inhibitors Regulate Retinoic Acid Receptor β Expression in Neuroblastoma Cells by Both Transcriptional and Posttranscriptional Mechanisms

Molecular Endocrinology, 2007

Research paper thumbnail of Combined effects of retinoic acid and histone deacetylase inhibitors on human neuroblastoma SH-SY5Y cells

Molecular Cancer Therapeutics, 2007

Research paper thumbnail of Acidic ribosomal proteins and histone H3 from Leishmania present a high rate of divergence

Memórias do Instituto Oswaldo Cruz, 2000

Research paper thumbnail of Anti-estrogenic actions of histone deacetylase inhibitors in MCF-7 breast cancer cells

Endocrine Related Cancer, 2007

Research paper thumbnail of Oil emulsion from Plukenetia huayllabambana (Sacha inchi) modifies nitric oxide and leptin in the liver and antioxidant and inflammation markers in the adipose tissue in obese rats

Functional Foods in Health and Disease, 2021

Background: Obesity is characterized by excessive accumulation of adipose tissue and is associate... more Background: Obesity is characterized by excessive accumulation of adipose tissue and is associated with higher risk of metabolic diseases and other comorbidities. Efficacious strategies including a diet high in “functional foods” are promising. Plukenetia huayllabambana known as Sacha Inchi (SI), is a legume which seeds are rich in proteins, tocopherols, and fatty acids such as omega-3 (ω-3). The latter has emerged as a potential protective nutrient against the cardiometabolic risks associated with obesity. Omega-3 changes the membrane lipid profile of hepatic and adipose cells triggering the expression of antioxidant and anti-inflammatory genes. However, there are few reports in relation to the effect of these oils in inflammatory and stress response related to obesity. In this sense, the present study evaluated the effect of SI oil emulsion on nitric oxide and leptin levels in the liver and some markers of oxidative stress and inflammation in adipose tissue from the rodent obesity...

Research paper thumbnail of An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections

PLoS neglected tropical diseases, 2016

Cutaneous and mucosal leishmaniasis is widely distributed in Central and South America. Leishmani... more Cutaneous and mucosal leishmaniasis is widely distributed in Central and South America. Leishmania of the Viannia subgenus are the most frequent species infecting humans. L. (V.) braziliensis, L. (V.) panamensis are also responsible for metastatic mucosal leishmaniasis. Conventional or real time PCR is a more sensitive diagnostic test than microscopy, but the cost and requirement for infrastructure and trained personnel makes it impractical in most endemic regions. Primary health systems need a sensitive and specific point of care (POC) diagnostic tool. We developed a novel POC molecular diagnostic test for cutaneous leishmaniasis caused by Leishmania (Viannia) spp. Parasite DNA was amplified using isothermal Recombinase Polymerase Amplification (RPA) with primers and probes that targeted the kinetoplast DNA. The amplification product was detected by naked eye with a lateral flow (LF) immunochromatographic strip. The RPA-LF had an analytical sensitivity equivalent to 0.1 parasites p...

Research paper thumbnail of Identificación de una nueva proteína en Leishmania (Viannia) peruviana

Revista peruana de medicina experimental y salud publica

Nucleotidic and aminoacidic sequence analysis from a clone of a phage λgt11 obtained from a expre... more Nucleotidic and aminoacidic sequence analysis from a clone of a phage λgt11 obtained from a expression vector library of Leishmania (Viannia) peruviana, established partial identity with the genes of the acidic ribosomal proteins P2 of Leishmania (Leishmania) infantum. These molecular findings along with certain conserved genomic domains, suggested by comparison of 14 sequences of eukaryotic P1 proteins, confirmed that the insert of the clone codes for acidic ribosomal protein P1 of L (V) peruviana refered as LpP1. This is the first report about this type of protein in Leishmania genus.

Research paper thumbnail of Outbreak of Cutaneous Leishmaniasis in Peruvian Military Personnel Undertaking Training Activities in the Amazon Basin, 2010

American Journal of Tropical Medicine and Hygiene, 2015

Military personnel deployed to the Amazon Basin are at high risk for cutaneous leishmaniasis (CL)... more Military personnel deployed to the Amazon Basin are at high risk for cutaneous leishmaniasis (CL). We responded to an outbreak among Peruvian Army personnel returning from short-term training in the Amazon, conducting active case detection, lesion sample collection, and risk factor assessment. The attack rate was 25% (76/303); the incubation period was 2-36 weeks (median = 8). Most cases had one lesion (66%), primarily ulcerative (49%), and in the legs (57%). Real-time polymerase chain reaction (PCR) identified Leishmania (Viannia) braziliensis (59/61 = 97%) and L. (V.) guyanensis (2/61 = 3%). Being male (risk ratio [RR] = 4.01; P = 0.034), not wearing long-sleeve clothes (RR = 1.71; P = 0.005), and sleeping in open rooms (RR = 1.80; P = 0.009) were associated with CL. Sodium stibogluconate therapy had a 41% cure rate, less than previously reported in Peru (∼70%; P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001). After emphasizing pre-deployment education and other basic prevention measures, trainees in the following year had lower incidence (1/278 = 0.4%; P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001). Basic prevention can reduce CL risk in deployed militaries.

Research paper thumbnail of Native Rodent Species Are Unlikely Sources of Infection for Leishmania (Viannia) braziliensis along the Transoceanic Highway in Madre de Dios, Peru

PLoS ONE, 2014

An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In ... more An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In Peru's Amazon region, the department of Madre de Dios (MDD) rises above the rest of the country in terms of the annual incidence rates of human leishmaniasis. Leishmania (Viannia) braziliensis is the species most frequently responsible for the form of disease that results in tissue destruction of the nose and mouth. However, essentially nothing is known regarding the reservoirs of this vectorborne, zoonotic parasite in MDD. Wild rodents have been suspected, or proven, to be reservoirs of several Leishmania spp. in various ecosystems and countries. Additionally, people who live or work in forested terrain, especially those who are not regionally local and whose immune systems are thus naïve to the parasite, are at most risk for contracting L. (V.) braziliensis. Hence, the objective of this study was to collect tissues from wild rodents captured at several study sites along the Amazonian segment of the newly constructed Transoceanic Highway and to use molecular laboratory techniques to analyze samples for the presence of Leishmania parasites. Liver tissues were tested via polymerase chain reaction from a total of 217 rodents; bone marrow and skin biopsies (ear and tail) were also tested from a subset of these same animals. The most numerous rodent species captured and tested were Oligoryzomys microtis (40.7%), Hylaeamys perenensis (15.7%), and Proechimys spp. (12%). All samples were negative for Leishmania, implying that although incidental infections may occur, these abundant rodent species are unlikely to serve as primary reservoirs of L. (V.) braziliensis along the Transoceanic Highway in MDD. Therefore, although these rodent species may persist and even thrive in moderately altered landscapes, we did not find any evidence to suggest they pose a risk for L. (V.) braziliensis transmission to human inhabitants in this highly prevalent region. Citation: Shender LA, De Los Santos M, Montgomery JM, Conrad PA, Ghersi BM, et al. (2014) Native Rodent Species Are Unlikely Sources of Infection for Leishmania (Viannia) braziliensis along the Transoceanic Highway in Madre de Dios, Peru. PLoS ONE 9(7): e103358.

Research paper thumbnail of Sensitivity and specificity of the real-time PCR assay compared to kDNA PCR stratified by site, type of sample and type of lesion

(*)<p>Statistically significant (<i>p</i><0.001) by Chi-square test.</p

Research paper thumbnail of Number of ear, tail, bone marrow, and liver tissues tested for <i>Leishmania</i> spp. via polymerase chain reaction shown by study site and species captured

1<p>Where n<sub>EC</sub> and n<sub>MZ</sub> are the total captures ... more 1<p>Where n<sub>EC</sub> and n<sub>MZ</sub> are the total captures at El Carmen and Mazuko, respectively</p>2<p><i>Proechimys spp</i>. included 1 <i>P. simonsi</i> at Mazuko and 6 <i>P. simonsi</i> at El Carmen. The remaining 11 at El Carmen were identified only to genus level.</p

Research paper thumbnail of <i>Leishmania</i> species identified by real-time PCR from individuals with suspected leishmaniasis from IMTAvH and HMC samples

(*)<p>No amplification for the <i>MPI</i> real-time PCR assay. <i>6PGD&lt... more (*)<p>No amplification for the <i>MPI</i> real-time PCR assay. <i>6PGD</i> real-time PCR results suggested infection by either <i>L.</i> (<i>V.</i>) <i>peruviana</i> or <i>L.</i> (<i>V.</i>) <i>braziliensis</i> but in the absence of MPI results, we could not discriminate between these two <i>Leishmania</i> species.</p

Research paper thumbnail of Upper bound Clopper-Pearson one-sided 95% confidence intervals for an observed <i>Leishmania</i> prevalence of 0%, evaluated by two strata: rodent species (for those species with ≥15 individuals represented) and study site

Research paper thumbnail of Genetic diversity and population structure of Leishmania (Viannia) braziliensis in the Peruvian jungle

PLOS Neglected Tropical Diseases

Background Human cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis is highly pr... more Background Human cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis is highly prevalent in the Peruvian jungle, where it affects military forces deployed to fight against drug trafficking and civilian people that migrate from the highland to the lowland jungle for economic activities such as mining, agriculture, construction, and chestnut harvest. We explored the genetic diversity and population structure of 124 L. (V.) braziliensis isolates collected from the highland (Junín, Cusco, and Ayacucho) and lowland Peruvian jungle (Loreto, Ucayali, and Madre de Dios). All samples were genotyped using Multilocus Microsatellite Typing (MLMT) of ten highly polymorphic markers. Principal findings High polymorphism and genetic diversity were found in Peruvian isolates of L. (V.) braziliensis. Most markers are not in Hardy-Weinberg equilibrium; this deviation is most likely caused by local inbreeding, as shown by the positive FIS values. Linkage Disequilibrium in subpopulations...

Research paper thumbnail of Diagnóstico molecular para Leishmaniasis

Revista Peruana de Medicina Experimental y Salud Pública, 1997

El potencial diagnóstico de epitopes inmunodominantes seleccionados fue ensayado satisfactoriamen... more El potencial diagnóstico de epitopes inmunodominantes seleccionados fue ensayado satisfactoriamente a fin de obtener una prueba serodiagnóstica alternativa para la Leishmaniasis Tegumentaria Americana. Dos proteínas recombinantes prometedoras de L. (v.) peruviana referidas como T-26-U2/T26-U4 fueron reconocidas por sueros individuales de pacientes con Leishmaniasis Tegumentaria Americana usando Western Blot. La sensibilidad de la prueba fue de 86% con sueros permanentes con Leishmaniasis peruana.

Research paper thumbnail of Running Title: Effects of HDACi and RA on SH-SY5Y cells

Combined effects of retinoic acid and histone deacetylase inhibitors on human neuroblastoma SH-SY... more Combined effects of retinoic acid and histone deacetylase inhibitors on human neuroblastoma SH-SY5Y cells

Research paper thumbnail of Inhibitors on Human Neuroblastoma

All-trans retinoic acid (RA) causes differentiation of neuroblastoma cells, and retinoids have be... more All-trans retinoic acid (RA) causes differentiation of neuroblastoma cells, and retinoids have been used in clinical trials in children with advanced neuroblastoma. Combination of RA with histone deacetylase inhibitors (HDACi) could result in improved antitumorigenic activity. We have examined the effect of the HDACi trichostatin A (TSA), sodium butyrate, and suberoylanilide hydroxamic acid (SAHA), alone and in combination with RA in human neuroblastoma SH-SY5Y cells. At concentrations that cause sustained increase of histone H3 acetylation, HDACi produced extensive apoptotic cell death as shown by flow cytometry analysis and induction of poly(ADP-ribose) polymerase proteolysis. HDACi inhibited SH-SY5Y cell growth at a much larger extent than RA. This compound did not cause apoptosis and did not further increase HDACimediated cell death. In contrast, both types of drugs cooperated to inhibit cell growth, although synergistic effects were not found. In surviving cells, HDACi repressed cyclin D1 expression and increased the cyclin kinase inhibitors (CKI) p21 Waf1/Cip1 and p27 Kip1. Cyclin D1 was not affected by RA, but this retinoid also increased CKI levels. Induction of p21 Waf1/Cip1 and p27 Kip1 by HDACi was further enhanced in the presence of RA. This effect seems to be at least partially due to transcriptional stimulation of CKI gene expression because both types of drugs cooperated to increase CKI mRNA levels and to activate the CKI promoters in transient transfection assays. These results show the strong antitumorigenic effects of HDACi in neuroblastoma cells and reinforce the idea that combination therapy could be useful to inhibit tumor growth.

Research paper thumbnail of Diagnóstico molecular para Leishmaniasis

Research paper thumbnail of Diagnostic Efficacy of Recombinase-Polymerase-Amplification Coupled with Lateral Flow Strip Reading in Patients with Cutaneous Leishmaniasis from the Amazonas Rainforest of Perú

Vector-Borne and Zoonotic Diseases, 2021

Cutaneous leishmaniasis (CL) is highly prevalent in rural and sylvatic regions of Latin America, ... more Cutaneous leishmaniasis (CL) is highly prevalent in rural and sylvatic regions of Latin America, with an estimated 55,000 annual cases. Diagnosis in resource-limited areas still relies on microscopy of dermal scrapings, while more sensitive methods like PCR are not attainable due to costs and lack of adequate health infrastructure. Isothermal amplification of Leishmania DNA can be performed without sophisticated equipment and training and may become a point of care (POC) test for health care centers with scarce resources. We evaluated the efficacy of recombinase-polymerase-amplification (RPA-LF) to diagnose CL in 226 patients attending a clinic in Puerto Maldonado within the Peruvian Amazon basin. Conventional PCR targeting kinetoplast DNA (kDNA-PCR) was used as the gold standard. Eight of 226 patients were considered true negatives (microscopy, kDNA-PCR, and RPA-LF negative), while RPA-LF resulted positive in 186 of 204 kDNA-PCR positive patients, yielding 91.2% (confidence interval [CI] = 86.5-94.4%) sensitivity and 93% (CI 88.6-95.8%) positive predictive value. There were 14% (32/226) discrepant samples alternating positive and negative results in similar proportions between both tests. Quantitative PCR used to resolve the discrepancies suggested that they occurred in samples with scarce parasite numbers as determined by high cycle threshold (Ct) values (≥32; cutoff 35.5). Microscopy had the lowest sensitivity of all methods (45.4%). Nested real-time PCR performed in 71 samples determined that Leishmania (Viannia) braziliensis was highly prevalent (69/71), and Leishmania (Viannia) lainsoni was present in only two isolates. Results indicated that RPA-LF has POC potential for CL endemic areas, yet further simplification and optimization coupled with field validation will be necessary to confirm its broad applicability.

Research paper thumbnail of Distribution and identification of sand flies naturally infected with Leishmania from the Southeastern Peruvian Amazon

PLoS neglected tropical diseases, 2017

Cutaneous leishmaniasis (CL) is an important health problem in the New World affecting civilian a... more Cutaneous leishmaniasis (CL) is an important health problem in the New World affecting civilian and military populations that are frequently exposed in endemic settings. The Peruvian region of Madre de Dios located near the border with Brazil is one of the most endemic CL regions in South America with more than 4,451 reported cases between 2010 and 2015 according to the Peruvian epidemiology directorate. However, little is known regarding the diversity and distribution of sand fly vectors in this region. In this study, we aimed to characterize the sand fly fauna in this endemic setting and identify sand fly species naturally infected with Leishmania possibly involved in pathogen transmission. Sand fly collections were carried out during 2014 and 2015 in the communities of Flor de Acre, Villa Primavera, Mavila and Arca Pacahuara using CDC light traps and Shannon traps. Collected specimens were identified and non-blood-fed females were selected for Leishmania infection screening using...

Research paper thumbnail of Histone Deacetylase Inhibitors Regulate Retinoic Acid Receptor β Expression in Neuroblastoma Cells by Both Transcriptional and Posttranscriptional Mechanisms

Molecular Endocrinology, 2007

Research paper thumbnail of Combined effects of retinoic acid and histone deacetylase inhibitors on human neuroblastoma SH-SY5Y cells

Molecular Cancer Therapeutics, 2007

Research paper thumbnail of Acidic ribosomal proteins and histone H3 from Leishmania present a high rate of divergence

Memórias do Instituto Oswaldo Cruz, 2000

Research paper thumbnail of Anti-estrogenic actions of histone deacetylase inhibitors in MCF-7 breast cancer cells

Endocrine Related Cancer, 2007

Research paper thumbnail of Oil emulsion from Plukenetia huayllabambana (Sacha inchi) modifies nitric oxide and leptin in the liver and antioxidant and inflammation markers in the adipose tissue in obese rats

Functional Foods in Health and Disease, 2021

Background: Obesity is characterized by excessive accumulation of adipose tissue and is associate... more Background: Obesity is characterized by excessive accumulation of adipose tissue and is associated with higher risk of metabolic diseases and other comorbidities. Efficacious strategies including a diet high in “functional foods” are promising. Plukenetia huayllabambana known as Sacha Inchi (SI), is a legume which seeds are rich in proteins, tocopherols, and fatty acids such as omega-3 (ω-3). The latter has emerged as a potential protective nutrient against the cardiometabolic risks associated with obesity. Omega-3 changes the membrane lipid profile of hepatic and adipose cells triggering the expression of antioxidant and anti-inflammatory genes. However, there are few reports in relation to the effect of these oils in inflammatory and stress response related to obesity. In this sense, the present study evaluated the effect of SI oil emulsion on nitric oxide and leptin levels in the liver and some markers of oxidative stress and inflammation in adipose tissue from the rodent obesity...

Research paper thumbnail of An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections

PLoS neglected tropical diseases, 2016

Cutaneous and mucosal leishmaniasis is widely distributed in Central and South America. Leishmani... more Cutaneous and mucosal leishmaniasis is widely distributed in Central and South America. Leishmania of the Viannia subgenus are the most frequent species infecting humans. L. (V.) braziliensis, L. (V.) panamensis are also responsible for metastatic mucosal leishmaniasis. Conventional or real time PCR is a more sensitive diagnostic test than microscopy, but the cost and requirement for infrastructure and trained personnel makes it impractical in most endemic regions. Primary health systems need a sensitive and specific point of care (POC) diagnostic tool. We developed a novel POC molecular diagnostic test for cutaneous leishmaniasis caused by Leishmania (Viannia) spp. Parasite DNA was amplified using isothermal Recombinase Polymerase Amplification (RPA) with primers and probes that targeted the kinetoplast DNA. The amplification product was detected by naked eye with a lateral flow (LF) immunochromatographic strip. The RPA-LF had an analytical sensitivity equivalent to 0.1 parasites p...

Research paper thumbnail of Identificación de una nueva proteína en Leishmania (Viannia) peruviana

Revista peruana de medicina experimental y salud publica

Nucleotidic and aminoacidic sequence analysis from a clone of a phage λgt11 obtained from a expre... more Nucleotidic and aminoacidic sequence analysis from a clone of a phage λgt11 obtained from a expression vector library of Leishmania (Viannia) peruviana, established partial identity with the genes of the acidic ribosomal proteins P2 of Leishmania (Leishmania) infantum. These molecular findings along with certain conserved genomic domains, suggested by comparison of 14 sequences of eukaryotic P1 proteins, confirmed that the insert of the clone codes for acidic ribosomal protein P1 of L (V) peruviana refered as LpP1. This is the first report about this type of protein in Leishmania genus.

Research paper thumbnail of Outbreak of Cutaneous Leishmaniasis in Peruvian Military Personnel Undertaking Training Activities in the Amazon Basin, 2010

American Journal of Tropical Medicine and Hygiene, 2015

Military personnel deployed to the Amazon Basin are at high risk for cutaneous leishmaniasis (CL)... more Military personnel deployed to the Amazon Basin are at high risk for cutaneous leishmaniasis (CL). We responded to an outbreak among Peruvian Army personnel returning from short-term training in the Amazon, conducting active case detection, lesion sample collection, and risk factor assessment. The attack rate was 25% (76/303); the incubation period was 2-36 weeks (median = 8). Most cases had one lesion (66%), primarily ulcerative (49%), and in the legs (57%). Real-time polymerase chain reaction (PCR) identified Leishmania (Viannia) braziliensis (59/61 = 97%) and L. (V.) guyanensis (2/61 = 3%). Being male (risk ratio [RR] = 4.01; P = 0.034), not wearing long-sleeve clothes (RR = 1.71; P = 0.005), and sleeping in open rooms (RR = 1.80; P = 0.009) were associated with CL. Sodium stibogluconate therapy had a 41% cure rate, less than previously reported in Peru (∼70%; P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001). After emphasizing pre-deployment education and other basic prevention measures, trainees in the following year had lower incidence (1/278 = 0.4%; P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001). Basic prevention can reduce CL risk in deployed militaries.

Research paper thumbnail of Native Rodent Species Are Unlikely Sources of Infection for Leishmania (Viannia) braziliensis along the Transoceanic Highway in Madre de Dios, Peru

PLoS ONE, 2014

An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In ... more An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In Peru's Amazon region, the department of Madre de Dios (MDD) rises above the rest of the country in terms of the annual incidence rates of human leishmaniasis. Leishmania (Viannia) braziliensis is the species most frequently responsible for the form of disease that results in tissue destruction of the nose and mouth. However, essentially nothing is known regarding the reservoirs of this vectorborne, zoonotic parasite in MDD. Wild rodents have been suspected, or proven, to be reservoirs of several Leishmania spp. in various ecosystems and countries. Additionally, people who live or work in forested terrain, especially those who are not regionally local and whose immune systems are thus naïve to the parasite, are at most risk for contracting L. (V.) braziliensis. Hence, the objective of this study was to collect tissues from wild rodents captured at several study sites along the Amazonian segment of the newly constructed Transoceanic Highway and to use molecular laboratory techniques to analyze samples for the presence of Leishmania parasites. Liver tissues were tested via polymerase chain reaction from a total of 217 rodents; bone marrow and skin biopsies (ear and tail) were also tested from a subset of these same animals. The most numerous rodent species captured and tested were Oligoryzomys microtis (40.7%), Hylaeamys perenensis (15.7%), and Proechimys spp. (12%). All samples were negative for Leishmania, implying that although incidental infections may occur, these abundant rodent species are unlikely to serve as primary reservoirs of L. (V.) braziliensis along the Transoceanic Highway in MDD. Therefore, although these rodent species may persist and even thrive in moderately altered landscapes, we did not find any evidence to suggest they pose a risk for L. (V.) braziliensis transmission to human inhabitants in this highly prevalent region. Citation: Shender LA, De Los Santos M, Montgomery JM, Conrad PA, Ghersi BM, et al. (2014) Native Rodent Species Are Unlikely Sources of Infection for Leishmania (Viannia) braziliensis along the Transoceanic Highway in Madre de Dios, Peru. PLoS ONE 9(7): e103358.