Juan Carlos Sepúlveda-Arias

Th1 and Th2 immune response to P30 and ROP18 peptides in human toxoplasmosis

Abstract We determined the specific lymphocyte proliferative response and cytokine profile production regarding Toxoplasma P30 (2017 from virulent and non-virulent strain) and ROP18 protein-derived peptides (from clonal lineages I, II and III) in 19 patients having ocular toxoplasmosis, five suffering chronic asymptomatic infection, nine with congenital toxoplasmosis and eight Toxoplasma negative people. A Beckman Coulter FC500 flow cytometer was used for determining antigen-specific T cells (CD3+ CD4+ or CD3+ CD8+ cells) in peripheral blood culture. IFN γ and IL10 levels were determined in culture supernatants. Specific CD4+ and CD8+ T cell response to total antigen and P30- and ROP18-derived peptides was observed in infected people. Ocular toxoplasmosis patients had a preferential Th2 response after antigenic stimulation. Non-virulent peptide 2017 was able to shift response toward Th1 in congenitally infected children and virulent peptide 2017 induced a Th2 response in chronically infected, asymptomatic people. An immune response in human toxoplasmosis after ex vivo antigenic stimulation was Th1- or Th2-skewed, depending on a patient’s clinical condition. Colombian ocular toxoplasmosis patients’ immune response was Th2-skewed, regardless of the nature of antigen stimulus.

Toxoplasmosis as a travel risk

Toxoplasma gondii is a protozoan parasite with worldwide distribution that infects more than one third of the global population. Primary infection in immunocompetent individuals is usually asymptomatic; however, different organs can be affected in immunocompromised individuals leading to the development of encephalitis, myocarditis or pneumonitis. The prevalence of infection with Toxoplasma as well as its genetic structure varies geographically and for that reason travel may be considered as a risk factor to acquire the infection. As toxoplasmosis is a foodborne disease, health care providers should give health education on prevention measures to all prospective travelers in order to decrease the risk of infection in endemic areas. This review presents an overview of the infection with T. gondii with some considerations for travelers to and from endemic zones.

β2‐adrenoreceptor polymorphisms in asthmatic and non‐asthmatic schoolchildren from colombia and their relationship to treatment response

Asthma is a chronic and recurrent disease. Its high prevalence around the world is the result of a complex interaction between genetic and environmental factors. The genetic aspects of susceptibility, severity, and response to treatment in asthma are of great scientific interest. The purpose of the study was to establish the relationship between the Gln27Glu and Arg16Gly alleles of the β2‐adrenergic receptor (ADRB2) gene with respect to the susceptibility to and severity of asthma, as well as the response to treatment in mestizo schoolchildren. 109 schoolchildren with asthma diagnosis and 137 asymptomatic controls were genotyped for the Arg16Gly and Gln27Glu alleles of the ADRB2 gene by minisequencing. Allele, genotype, and haplotype frequencies of the ADRB2 gene between asthmatic and non‐asthmatic as well as demographic, clinical, and spirometric variables among asthmatic patients according to their genotype were compared. ADRB2 gene expression was determined by real‐time quantitative PCR. No statistical differences were found in allele, genotype, and haplotype frequencies of the ADRB2 gene between cases and controls. We did not find differences between asthmatic patients classified according to their ADRB2 genotypes and haplotypes when evaluating demographic, clinical, and spirometric variables. The ADRB2 genotype and haplotype are not associated with spirometric responses or ADRB2 gene expression after administration of a β‐2 agonist plus a glucocorticoid. These results suggest that in the group of mestizo schoolchildren studied, the Arg16Gly and Gln27Glu polymorphisms are not markers of susceptibility or severity of asthma and do not affect ADRB2 gene expression during the rescue therapy. Pediatr Pulmonol. 2012. 47:848–855.

Comparison of five protocols to extract DNA from paraffin-embedded tissues for the detection of human papillomavirus

Formalin-fixed paraffin-embedded (FFPE) tissues are a valuable source of DNA with which to perform large retrospective studies on the epidemiology of HPV infection. Five different DNA extraction protocols were carried out to evaluate the DNA obtained from FFPE samples with polymerase chain reaction (PCR) using two primer sets to amplify a constitutive human gene, β-globin, and two primer sets to detect the L1 and E6 HPV genes. From the five DNA extraction protocols evaluated, the best results were obtained with protocol A, corresponding to a crude extract from the sample. With the procedures described herein, we were able to amplify DNA extracted from archival paraffin blocks stored for six years. However, the amplification products were more efficiently obtained with primers that amplified shorter fragments. This result indicates that a major factor limiting the extraction process in these samples is DNA fragmentation, a factor that will naturally vary between the different specimens evaluated. Also, depending upon the extraction method, PCR amplification of a human gene does not necessarily guarantee the successful extraction of viral DNA. In conclusion, different DNA and HPV detection methods can significantly influence the results. Therefore, the DNA extraction methods and primers used for DNA amplification in fixed tissues need to be chosen carefully, depending on the specific requirements of the study being carried out.

Hepatitis B and C prevalence among heroin addicts in methadone maintenance treatment (MMT) and not in MMT in Pereira, Colombia

This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. In a recent systematic review, Nelson et al showed that injection drug abuse is an important factor associated with a high prevalence of hepatitis C virus (HCV) and hepatitis B virus (HBV) infection [1]. In that review, only data from Argentina, Brazil, Mexico, Paraguay and Uruguay were included, due to the lack of information on other countries in Latin America and in the Caribbean. After an extensive literature search about the prevalence of injection drug use in Colombia, only four reports were found on three different Colombian cities. The prevalence of intravenous heroin administration in Medellin ranges between 0.2% in a prison population (VESPA Survey reported by Castaño) [2] and 24% in drug addiction treatment centers [3]. However, a comparison between both studies cannot be carried out due to differences in sample size, methodology and population evaluated. In Bucaramanga, the prevalence of intravenous illegal drug use in three drug rehabilitation centers and one prison was 4.2% [4]. Finally, a survey carried out on 895 inhabitants from Pereira showed a prevalence of intravenous drug use of 26.9%, including the use of heroin and cocaine [5] (Figure 1). Hepatitis C virus (HCV) testing and counseling are important strategies to potentially reduce HCV transmission in young adult injection drug users [6]. However, in developing countries, the health systems underappreciate the role of injection drug users (IDU) in the transmission of infectious agents such as HCV, hepatitis B virus (HBV) and human immunodeficiency virus (HIV). Moreover, the treatment of HCV infection in IDU is still low in developing countries due to patient-and system-related barriers. Although HCV infection is a public health problem around the world and chronic viral hepatitis are the major etiological factors of end-stage liver diseases, in Colombia there are no studies of HCV and HBV prevalence in IDU. The prevalence of HCV in multi-transfused patients is 9% [7], whereas a more recent study carried out in 697 inhabitants from four different states showed a frequency of anti-HCV markers of 3.55% [8]. On the other hand, the prevalence of HBV in Colombia ranges between 1.97 and 8.39% in several regions [8-10]. As part of an ongoing work evaluating the immune status of IDU and a control population in Pereira, Colombia (approved by the …

Anti-inflammatory effects of the main constituents and epoxides derived from the essential oils obtained from Tagetes lucida, Cymbopogon citratus, Lippia alba and Eucalyptus citriodora

The aim of this study was to evaluate the anti-inflammatory activities of the main constituents and epoxides derived from the essential oils (EOs) obtained from Tagetes lucida, Cymbopogon citratus, Lippia alba and Eucalyptus citriodora. The main constituents were isolated and then epoxidized with dimethyldioxirane (DMDO) in an acetone solution. The anti-inflammatory effects were studied in vitro on murine macrophages stimulated with bacterial lipopolysaccharide. All tested samples inhibited both nitric oxide (NO) and prostaglandin E2 (PGE2) production, but the EO from T. lucida and carvone exhibited the highest inhibitory effects. Most of the studied samples altered the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). The studied epoxides inhibited both NO and PGE2 production, but at the RNA level, they primarily inhibited COX-2. These results show, for the first time, the anti-inflammatory effects of epoxides generated from the main components of EOs from Colombian plants.

Burden of Human Papillomavirus Infection in Latin America

Human papillomavirus (HPV)-associated disease is an important public health problem worldwide. HPV is not only associated with 530,000 new cases of cervical cancer and 275,000 cervical cancer deaths worldwide each year but also causes vulvar, vaginal, anal, and penile cancers as well as cervical and vulvar/vaginal precancerous lesions, genital warts, and respiratory papillomatosis. Most individuals are not aware that they are infected with HPV because of its subclinical or asymptomatic presentation, and thus, the virus can be spread easily and unknowingly during sexual foreplay or sexual intercourse [1].

Plant expression systems, a budding way to confront chikungunya and Zika in developing countries?

Plant expression systems could be used as biofactories of heterologous proteins that have the potential to be used with biopharmaceutical aims and vaccine design. This technology is scalable, safe and cost-effective and it has been previously proposed as an option for vaccine and protein pharmaceutical development in developing countries. Here we present a proposal of how plant expression systems could be used to address Zika and chikungunya outbreaks through development of vaccines and rapid diagnostic kits.

Genetic Polymorphisms in Cytokine Genes in Colombian Patients with Ocular Toxoplasmosis

ABSTRACT Toxoplasmosis is caused by infection with the protozoan parasite Toxoplasma gondii, which has the capacity to infect all warm-blooded animals worldwide. Toxoplasmosis is a major cause of visual defects in the Colombian population; however, the association between genetic polymorphisms in cytokine genes and susceptibility to ocular toxoplasmosis has not been studied in this population. This work evaluates the associations between polymorphisms in genes coding for the cytokines tumor necrosis factor alpha (TNF-α) (rs1799964, rs1800629, rs1799724, rs1800630, and rs361525), interleukin 1β (IL-1β) (rs16944, rs1143634, and rs1143627), IL-1α (rs1800587), gamma interferon (IFN-γ) (rs2430561), and IL-10 (rs1800896 and rs1800871) and the presence of ocular toxoplasmosis (OT) in a sample of a Colombian population (61 patients with OT and 116 healthy controls). Genotyping was performed with the “dideoxynucleotide (ddNTP) primer extension” technique. Functional-effect predictions of single nucleotide polymorphisms (SNPs) were done by using FuncPred. A polymorphism in the IL-10 gene promoter (−1082G/A) was significantly more prevalent in OT patients than in controls (P = 1.93e−08; odds ratio [OR] = 5.27e+03; 95% confidence interval [CI] = 3.18 to 8.739; Bonferroni correction [BONF] = 3.48e−07). In contrast, haplotype “AG” of the IL-10 gene promoter polymorphisms (rs1800896 and rs1800871) was present at a lower frequency in OT patients (P = 7e−04; OR = 0.10; 95% CI = 0.03 to 0.35). The +874A/T polymorphism of IFN-γ was associated with OT (P = 3.37e−05; OR = 4.2; 95% CI = 2.478 to 7.12; BONF = 6.07e−04). Haplotype “GAG” of the IL-1β gene promoter polymorphisms (rs1143634, rs1143627, and rs16944) appeared to be significantly associated with OT (P = 0.0494). The IL-10, IFN-γ, and IL-1β polymorphisms influence the development of OT in the Colombian population.

Inflammatory Markers in the Staging of Bipolar Disorder: A Systematic Review of the Literature

BACKGROUND Previous studies suggest that inflammatory molecules play an important role in the pathophysiology of Bipolar Disorder (BD). The evidence suggests that BD may present a progressive course. Therefore there are theories that postulate the relationship between progression and stages of the disease with distinct peripheral biomarkers. OBJECTIVE The aim of this study was to carry out a systematic review of the literature of studies about the association between peripheral inflammatory markers and clinical variables related with staging in BD patients. METHODS We conducted a systematic review using electronic databases: PubMed, SciELO, LiLACS and PsycINFO. Keywords were divided into inflammatory markers and, BD and staging. Studies involving euthymic BD patients, studies evaluating peripheral biomarkers and studies correlating these with clinical variables related to neuroprogression or stage of BD were included. RESULTS We present and discuss the methods and findings of ten articles. The inflammatory markers were measured with different techniques and show some contradictories results. The TNF superfamily and inflammatory cytokines may have a relationship with the neuroprogression of the disease. CONCLUSIONS This study suggests that TNF and ILs could play a role in neuroprogression. However, longitudinal studies are needed to clarify the relationship between factors associated with neuroprogression.