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Throughout its lifecycle, Entamoeba histolytica encounters a variety of stressful conditions. This parasite possesses Heat Shock Response Elements (HSEs) which are crucial for regulating the expression of various genes, aiding in its adaptation and survival. These HSEs are regulated by Heat Shock Transcription Factors (EhHSTFs). Our research has identified seven such factors in the parasite, designated as EhHSTF1 through to EhHSTF7. Significantly, under heat shock conditions and in the presence of the antiamoebic compound emetine, EhHSTF5, EhHSTF6, and EhHSTF7 show overexpression, highlighting their essential role in gene response to these stressors. Currently, only EhHSTF7 has been confirmed to recognize the HSE as a promoter of the EhPgp5 gene (HSE_EhPgp5), leaving the binding potential of the other EhHSTFs to HSEs yet to be explored. Consequently, our study aimed to examine, both in vitro and in silico, the oligomerization, and binding capabilities of the recombinant EhHSTF5 protein (rEhHSTF5) to HSE_EhPgp5. The in vitro results indicate that the oligomerization of rEhHSTF5 is concentration-dependent, with its dimeric conformation showing a higher affinity for HSE_EhPgp5 than its monomeric state. In silico analysis suggests that the alpha 3 α-helix (α3-helix) of the DNA-binding domain (DBD5) of EhHSTF5 is crucial in binding to the major groove of HSE, primarily through hydrogen bonding and salt-bridge interactions. In summary, our results highlight the importance of oligomerization in enhancing the affinity of rEhHSTF5 for HSE_EhPgp5 and demonstrate its ability to specifically recognize structural motifs within HSE_EhPgp5. These insights significantly contribute to our understanding of one of the potential molecular mechanisms employed by this parasite to efficiently respond to various stressors, thereby enabling successful adaptation and survival within its host environment.
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Membro 1 da Subfamília B de Cassetes de Ligação de ATP , Entamoeba histolytica , Regiões Promotoras Genéticas , Proteínas de Protozoários , Sítios de Ligação , Simulação por Computador , Entamoeba histolytica/genética , Entamoeba histolytica/metabolismo , Resposta ao Choque Térmico/genética , Ligação Proteica , Multimerização Proteica , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/química , Elementos de Resposta , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/química , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismoRESUMO
Resumen Antecedentes: El incremento seminal de especies reactivas de oxígeno (ERO) se ha vinculado con la oxidación del ADN y anormalidades en la morfología espermática. La enzima 8-oxoguanina ADN glucosilasa 1 (OGG1) repara la oxidación del ADN. Sin embargo, la presencia del polimorfismo en la OGG1 que involucra cambio de citocina (C) por guanina (G), resultando la sustitución de una cisteína por una serina en el codón 326 (Ser326Cis), ha demostrado disminución en la reparación del ADN oxidado. Estudios del polimorfismo Ser326Cis en hombres españoles y asiáticos con infertilidad demostraron que el alelo G(Cis) incrementa las ERO, impactando en la infertilidad y en la teratozoospermia. Objetivo: Conocer la prevalencia del polimorfismo Ser326Cis de OGG1 en pacientes con teratozoospermia. Método: Se analizaron parámetros espermáticos y el polimorfismo Ser326Cis de OGG1 de 81 muestras de semen con teratozoospermia. Resultados: Los genotipos detectados fueron Ser326Ser(CC) 43%, Ser326Cis(CG) 41% y Cis326Cis(GG) 16%. La frecuencia del alelo G(Cis) fue de 0.4, valor mayor a la frecuencia reportada en las bases de datos disponibles para poblaciones americanas (0.21-0.29), los parámetros espermáticos no se relacionaron con el polimorfismo Ser326Cis. Conclusión: El alelo G(Cis) es un factor que contribuye a la infertilidad.
Abstract Background: The increase in seminal reactive oxygen species (ROS) has been linked to DNA oxidation and abnormalities in sperm morphology. The enzyme 8-oxoguanine DNA glycosylase 1 (OGG1) repairs DNA oxidation. However, the presence of the polymorphism in OGG1 that involves a change of cytokine (C) to guanine (G) resulting in the substitution of a cystine for serine at codon 326 (Ser326Cys) has shown a decrease in the repair of oxidized DNA. Studies of the Ser326Cys polymorphism in Spanish and Asian men with infertility demonstrated that the G(Cys) allele increases ROS, impacting infertility and teratozoospermia. Objective: To know the prevalence of the Ser326Cys polymorphism of OGG1 in patients with teratozoospermia. Method: Sperm parameters and the Ser326Cys polymorphism of OGG1 were analyzed from 81 semen samples. Results: The genotypes detected were Ser326Ser(CC) 43%, Ser326Cys(CG) 41%, and yis326Cys(GG) 16%. The frequency of the G allele (Cys) was 0.4, a value higher than the frequency reported in the databases available for American populations (0.21-0.29), the sperm parameters were not related to the Ser326Cys polymorphism. Conclusion: The G (Cys) allele is a factor that contributes to infertility.
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ABSTRACT With an estimated approximately 2 million deaths per year, diabetes is one of the top 5 deadliest noncommunicable diseases globally. Although this disease is not fatal, the degradation of the patient's health due to a bad plan to control their glucose levels can have a fatal outcome. In order to lay the foundations for the development of a device that allows estimating glucose levels in some body fluid, we present the results obtained not only for the estimation of glucose in deionized water, but also describe the development and configuration of the created device. After analyzing 50 signals obtained from 5 different glucose concentrations, the feasibility of using the developed device for the analysis is evident, since, considering the K-Nearest Neighbors (KNN) algorithm, all the signals were associated correctly to the glucose group to which they belong.
RESUMEN Con un estimado de aproximadamente 2 millones de muertes por año, la diabetes es una de las 5 enfermedades no transmisibles más mortales a nivel mundial. Aunque esta enfermedad no es mortal, el deterioro de la salud del paciente por un mal plan para controlar sus niveles de glucosa puede tener un desenlace fatal. Con el fin de sentar las bases para el desarrollo de un dispositivo que permita estimar los niveles de glucosa en algún fluido corporal, presentamos los resultados obtenidos no solo para la estimación de glucosa en agua desionizada, sino que también describimos el desarrollo y configuración del dispositivo creado. Luego de analizar 50 señales obtenidos a partir de 5 concentraciones de glucosa diferentes, se evidencia la factibilidad de utilizar el dispositivo desarrollado para el análisis, ya que, considerando el algoritmo K-Nearest Neighbors (KNN), todas las señales se asociaron correctamente al grupo de glucosa al que pertenecen.
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Amoebiasis is produced by the parasite Entamoeba histolytica; this disease affects millions of people throughout the world who may suffer from amoebic colitis or amoebic liver abscess. Metronidazole is used to treat this protozoan, but it causes important adverse effects that limit its use. Studies have shown that riluzole has demonstrated activity against some parasites. Thus, the present study aimed, for the first time, to demonstrate the in vitro and in silico anti-amoebic activity of riluzole. In vitro, the results of Entamoeba histolytica trophozoites treated with IC50 (319.5 µM) of riluzole for 5 h showed (i) a decrease of 48.1% in amoeba viability, (ii) ultrastructural changes such as a loss of plasma membrane continuity and alterations in the nuclei followed by lysis, (iii) apoptosis-like cell death, (iv) the triggering of the production of reactive oxygen species and nitric oxide, and (v) the downregulation of amoebic antioxidant enzyme gene expression. Interestingly, docking studies have indicated that riluzole presented a higher affinity than metronidazole for the antioxidant enzymes thioredoxin, thioredoxin reductase, rubrerythrin, and peroxiredoxin of Entamoeba histolytica, which are considered as possible candidates of molecular targets. Our results suggest that riluzole could be an alternative treatment against Entamoeba histolytica. Future studies should be conducted to analyze the in vivo riluzole anti-amoebic effect on the resolution of amebic liver abscess in a susceptible model, as this will contribute to developing new therapeutic agents with anti-amoebic activity.
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Resumen OBJETIVO: Determinar la frecuencia del alelo Ala en una muestra de mujeres mexicanas con diabetes mellitus gestacional y asociar su repercusión en la glucemia. MATERIALES Y MÉTODOS: Estudio ambispectivo, observacional, transversal y correlacional efectuado en una cohorte de pacientes con diabetes gestacional atendidas entre los meses de enero a junio del 2014 en el Hospital Militar de Especialidades de la Mujer y Neonatología de la Secretaría de la Defensa Nacional en la Ciudad de México. Se evaluó el polimorfismo mediante amplificación de un fragmento de ADN mediante la reacción en cadena de la polimerasa (PCR) y su secuenciación. RESULTADOS: Se estudiaron 81 pacientes; 3 de ellas con el alelo Ala, con concentraciones de glucosa menores y antecedente de más abortos en comparación con las mujeres sin el alelo Ala. CONCLUSIONES: La coexistencia del alelo Ala en mujeres embarazadas con diagnóstico de diabetes mellitus gestacional pudiera tener un efecto protector en contra de la hiperglucemia en el embarazo y el riesgo de aborto.
Abstract OBJECTIVE: To determine the frequency of peroxisomal proliferator-activated receptor gamma (PPARg) polymorphism of proline substituted with an alanine in amino acid 12 (Pro12Ala), in women with gestational diabetes mellitus and associate its impact with glycemia. MATERIALS AND METHODS: An ambispective, observational, cross-sectional and correlational study was carried out in a cohort of women with gestational diabetes that included 81 pregnant women treated at the Military Hospital for Women's Specialties and Neonatology of the Ministry of National Defense in the city from Mexico. Polymorphism was evaluated by amplification of a DNA fragment by PCR Polymerase Chain Reaction and its sequencing. RESULTS: The results indicated that 13.5% of the women carriers of the Ala allele also had lower blood glucose values and a history with a higher number of abortions compared to women without the Ala allele. CONCLUSIONS: The presence of the Ala allele in pregnant women with gestational diabetes mellitus could have a protective effect against hyperglycemia in pregnancy and a risk of abortion.
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Various immunopathological events characterize the systemic acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Moreover, it has been reported that coronavirus disease 2019 (COVID-19) vaccination and infection by SARS-CoV-2 induce humoral immunity mediated by B-cell-derived antibodies and cellular immunity mediated by T cells and memory B cells. Immunoglobulins, cytokines, and chemokines play an important role in shaping immunity in response to infection and vaccination. Furthermore, different vaccines have been developed to prevent COVID-19. Therefore, this research aimed to analyze and compare Fourier-transform infrared (FTIR) spectra of vaccinated people with a positive (V-COVID-19 group) or negative (V-Healthy group) real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) test, evaluating the immunoglobulin and cytokine content as an immunological response through FTIR spectroscopy. Most individuals that integrated the V-Healthy group (88.1%) were asymptomatic; on the contrary, only 28% of the V-COVID-19 group was asymptomatic. Likewise, 68% of the V-COVID-19 group had at least one coexisting illness. Regarding the immunological response analyzed through FTIR spectroscopy, the V-COVID-19 group showed a greater immunoglobulins G, A, and M (IgG, IgA, and IgM) content, as well as the analyzed cytokines interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-É), and interleukins 1ß, 6, and 10 (IL-1ß, IL-6, and IL-10). Therefore, we can state that it was possible to detect biochemical changes through FTIR spectroscopy associated with COVID-19 immune response in vaccinated people.
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COVID-19 , SARS-CoV-2 , Humanos , Espectroscopia de Infravermelho com Transformada de Fourier , Citocinas , Imunidade HumoralRESUMO
BACKGROUND: Cytochrome P4502E1 (CYP2E1) metabolizes environmental toxins, however, compound metabolism can produce oxidative stress, causing in-cell toxicity and sometimes transformation. AIM: To evaluate CYP2E1 gene expression and its effects in antioxidant defenses, and cell toxicity in printing workers. METHODS: The hierarchical method of health and chemical risk was used to evaluate chemical exposure in workplace. Blood samples and buccal epithelial cells were obtained from printing workers, and workers without any history of occupational exposure to chemicals (control group). Gene expression of CYP2E1, and antioxidant enzymes Superoxide dismutase (SOD) and Catalase (CAT) from leukocytes were evaluated. Hematic analysis and cell-free DNA from plasma were analyzed. Frequencies of cells with micronuclei (MN) and nuclear abnormalities from buccal epithelial cells were explored. RESULTS: Evaluation of chemical exposure in working place demonstrated that ethyl alcohol, isopropyl alcohol, and isophorone represent 91% of the accumulated potential risk. CYP2E1 expression showed a 2.5-fold overexpression in the printing workers compared to the control group. SOD expression showed a 0.5-fold lower level in the printing workers than the control group, and CAT expression showed no differences between groups. Lower red blood cell and platelet values were detected in the printing workers than in the control group, and cell-free DNA plasma concentration was 3-fold higher in the printing workers than in the control group. The printing workers showed a higher frequency of cells with MN and nuclear anomalies than the control group. CONCLUSION: CYP2E1 overexpression triggers antioxidant defenses and toxic cell effects in printing workers.
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Ácidos Nucleicos Livres , Exposição Ocupacional , Citocromo P-450 CYP2E1/genética , Antioxidantes/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Estresse Oxidativo/genética , Exposição Ocupacional/efeitos adversos , Exposição Ocupacional/análise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ácidos Nucleicos Livres/metabolismo , Impressão TridimensionalRESUMO
High-risk human papillomavirus (HR-HPV) infection, followed by chronic inflammation and oxidative stress, is a major risk factor of male infertility. In this study, we explored the potential impact of high-risk (HR) HPV genotypes in single infection (SI) and multiple infections (MI) that promote CYP2E1 expression, oxidative damage and pro-inflammatory cytokines, possibly contributing to sperm damage and male infertility. Semen samples from 101 infertile military men were studied. We analyzed seminal parameters, namely, HPV genotyping, cytochrome P450 2E1 (CYP2E1), oxidative stress biomarkers (total antioxidant capacity (TAC), catalase (CAT) and superoxide dismutase (SOD)), lipid peroxidation (LPO), 8-hydroxiguanosine (8-OHdG) and pro-inflammatory cytokines (IFN-γ, IL-1ß, IL-4, IL-6 and IL-8). Eighty-one men (80.2%, 81/101) were positive for HPV infection, and MI-HR-HPV was higher than SI-HR-HPV (63% vs. 37%). HPV-52 was the most frequently detected type (18.5%), followed by HPV-33 (11.1%), and the most frequent combination of genotypes detected was HPV-33,52 (11.1%), followed by HPV-18,31 (6.2%). The group with infected samples presented lower normal morphology and antioxidant levels compared to non-infected samples. In concordance, the infected group showed high levels of LPO, IFN-γ, IL-1ß, IL-4 and IL-6 and downregulation of CAT and SOD enzymes. Interestingly, changes in motility B, low levels of TAC, overexpression of CYP2E1, LPO and IL-8 levels were higher in MI-HR-HPV than SI-HR-HPV, suggesting that HPV infection promotes a chronic inflammatory process and a toxic and oxidative microenvironment, which increases with MI-HPV infections.
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Background and Objectives: To investigate the effect of infection with human papillomavirus (HPV) or Chlamydia trachomatis (CT) and HPV + CT coinfection on sperm quality, inflammation, and the state of oxidative stress (OS) in asymptomatic infertile men. Materials and Methods: Semen samples from 84 asymptomatic military infertile men were studied. The polymerase chain reaction (PCR) was used for the molecular detection of HPV and CT. Semen parameters were analyzed according to the World Health Organization guidelines. Inflammation was evaluated by an IL-1ß, IL-6, and IFN-γ enzyme-linked immunosorbent assay (ELISA) and OS by the quantification of lipid peroxidation (LPO), 8-hydroxydeoxyguanosine (8-OHdG), and total antioxidant capacity (TAC). Results: A total of 81 of the 84 (96.4%) samples were positives for the pathogens, with 55/81 (68%) being positive for HPV, 11/81 (13.5%) for CT, and 15/81 (18.5%) for HPV + CT coinfection. Seminal parameters were affected in the infected groups, including pH increases above the normal range in all groups. An abnormal sperm morphology was observed in the HPV and HPV + CT groups. Higher cytokine levels were detected in the HPV group and the highest IL-1ß level was found in the HPV + CT group. No cytokines were detected in the CT group. High LPO and 8-OHdG levels were found in all groups with a lower TAC. Comparisons between groups showed the highest OS state was observed in the HPV group. Conclusions: High HPV infection or coinfection (HVP + CT) in these infertile men suggest compromising male fertility by inducing a proinflammatory state and OS. Infection with CT suggests an alteration of the state of OS by promoting an alkaline pH.
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Alphapapillomavirus , Coinfecção , Infertilidade Masculina , Infecções por Papillomavirus , Chlamydia trachomatis , Humanos , Masculino , Estresse Oxidativo , Papillomaviridae , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/epidemiologia , EspermatozoidesRESUMO
INTRODUCTION: Infection and inflammation of the reproductive tract by Chlamydia trachomatis (CT) are recognized as significant risk factors for male infertility. This study aimed to evaluate CT infection and its effects on seminal parameters and cytokines in asymptomatic patients with teratozoospermia. MATERIAL AND METHODS: Semen samples from one hundred four male patients were collected, and CT detection was performed by polymerase chain reaction (PCR). The quality (volume, sperm concentration, pH, motility, morphology, and leucocytes) of the semen was measured by standard procedures recommended by the World Health Organization (WHO). Pro-inflammatory cytokines [interleukin (IL)-1 ß, IL-6, IL-8, tumor necrosis factor α (TNF-α), and interferon γ (IFN-γ)], as well as anti-inflammatory cytokines (IL-4, IL-10), were determined by using enzyme-linked immunosorbent assay (ELISA). The frequency of CT infection was expressed as a percentage. Descriptive statistics were used for comparison of cytokines from infertile men, and then the Mann-Whitney U test was applied for the contrast of seminal parameters and cytokines from CT-infected versus non-CT infected men. RESULTS: A ratio of 33/104 (31.7%) patients were positive for CT infection. The ejaculate of positive CT infection was found to have increased pH (pH = 7.65 in non-CT infected vs. 7.94 CT-infected men; p = 0.026). High levels of pro-inflammatory cytokines were found in the population studied; however, infected males were noted to have high levels of IL-1 ß [184.66 (0-3985.33 pg/ml), p = 0.001] and IL-6 [87.8 (0-1042.8 pg/ml), p = 0.001]. CONCLUSIONS: CT infection increased seminal pH, as well as IL-1 ß and IL-6 cytokines, suggesting a potential role of infection and inflammation in asymptomatic patients with teratozoospermia.
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OBJECTIVE: The purpose of the current study was to evaluate the modulation of Calcitonin gene-related peptide (CGRP) associated to the efficacy of Electroacupuncture (EA) in the reduction of climacteric symptoms. METHODS: Nine women between 51 and 59 years old with climacteric syndrome in menopause or perimenopause were included. Patients with hormone replacement therapy, psychiatric treatment with antidepressants, or acupuncture treatment in the last 3 months were excluded. A 4 Hz EA treatment was performed at acupoints Shenshu (BL-23), Pishu (BL-20), Guanyuan (REN-4), Taixi (KID-3), Fuliu (KID-7), Sanyinjiao (SP-6) and Neiguan (P-6) points. Women were treated two times a week for five consecutive weeks for a total treatment of 10 sessions. The menopause rating scale (MRS) was used to evaluate symptoms reduction and CGRP gene expression was measured before and after 10 EA session. RESULTS: The results shown that climacteric symptoms diminish significantly after EA therapy. CGRP gene expression was down-regulated, evidencing a decrease of 5-fold after EA therapy respect to the initial condition. CONCLUSION: EA treatment was associated with improvement in patients with climacteric syndrome and may be related to modulation of CGRP levels.
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Terapia por Acupuntura , Climatério , Eletroacupuntura , Pontos de Acupuntura , Peptídeo Relacionado com Gene de Calcitonina , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: To evaluate the pulmonary alterations in workers from a dental prosthesis laboratory and explore dust and bacterial dissemination generated in the laboratory. METHODS: Spirometry and computerized axial tomography were performed on 67 workers. Dust in workplace air was determined using the filtration-gravimetric method, and bacterial detection was explored using 16S rDNA gene sequencing. RESULTS: Pulmonary alterations were detected in 37% of the workers. Airborne dust concentrations were determined to be higher than the maximum permissible exposure limit, and bacterial detection analysis revealed 23 bacterial genera. The most frequently detected bacterial genus was Sphingomonas sp., which has been described as microbiota associated with disease of the oral cavity. CONCLUSIONS: The results of this study highlight the importance of the implementation of biosecurity measures, improvement of ventilation systems, and routine disinfection of dental impressions.
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Poluentes Ocupacionais do Ar , Prótese Dentária , Exposição Ocupacional , Local de Trabalho , Poluentes Ocupacionais do Ar/análise , Poeira/análise , Humanos , Laboratórios , Exposição Ocupacional/análiseRESUMO
Naegleria fowleri produces a fatal disease called primary amebic meningoencephalitis (PAM), which is characterized by an extensive inflammatory reaction in the CNS. It is known that the immune response is orchestrated mainly by neutrophils, which activate several defense mechanisms in the host, including phagocytosis, the release of different enzymes such as myeloperoxidase (MPO), and the production of neutrophil extracellular traps. However, the mechanisms by which amoebas evade the neutrophil response are still unknown. In this study, we analyzed the ability of N. fowleri to respond to the stress exerted by MPO. Interestingly, after the interaction of trophozoites with neutrophils, the amoeba viability was not altered; however, ultrastructural changes were observed. To analyze the influence of MPO against N. fowleri and its participation in free radical production, we evaluated its enzymatic activity, expression, and localization with and without the specific 4-aminobenzoic acid hydrazide inhibitor. The production of oxidizing molecules is the principal mechanism used by neutrophils to eliminate pathogens. In this context, we demonstrated an increase in the production of NO, superoxide anion, and reactive oxygen species; in addition, the overexpression of several antioxidant enzymes present in the trophozoites was quantified. The findings strongly suggest that N. fowleri possesses antioxidant machinery that is activated in response to an oxidative environment, allowing it to evade the neutrophil-mediated immune response, which may contribute to the establishment of PAM.
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Interações Hospedeiro-Parasita/imunologia , Naegleria fowleri/metabolismo , Neutrófilos/fisiologia , Oxirredutases/biossíntese , Peroxidase/fisiologia , Proteínas de Protozoários/biossíntese , Compostos de Anilina/farmacologia , Animais , Forma Celular , Grânulos Citoplasmáticos/enzimologia , Grânulos Citoplasmáticos/ultraestrutura , Indução Enzimática , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Naegleria fowleri/enzimologia , Naegleria fowleri/crescimento & desenvolvimento , Naegleria fowleri/ultraestrutura , Neutrófilos/efeitos dos fármacos , Óxido Nítrico/metabolismo , Oxirredução , Estresse Oxidativo , Oxirredutases/genética , Peroxidase/antagonistas & inibidores , Proteínas de Protozoários/genética , Espécies Reativas de Oxigênio , Superóxidos/metabolismo , Vacúolos/ultraestruturaRESUMO
The plasma membrane is essential in the pathogenicity of several microorganisms. However, to date, there are few studies related to the plasma membrane proteins in Naegleria fowleri; this amoeba produces a fatal disease called primary amoebic meningoencephalitis. In the present study, we analyzed the electrophoretic pattern of the membrane proteins of N. fowleri and compared it with the nonpathogenic N. lovaniensis and N. gruberi. We detected a 23-kDa protein (Nf23) present at a higher level in N. fowleri than in the nonpathogenic amoebae. The mass spectrometry analysis showed that the Nf23 protein has a sequence of 229 amino acids that corresponds to a membrane protein. The mRNA level of nf23 was overexpressed 4-fold and 40,000-fold in N. fowleri compared with N. lovaniensis and N. gruberi, respectively. Moreover, we found a 5-fold overexpression of nf23 in N. fowleri trophozoites recovered from mouse brains compared with trophozoites axenically cultivated. In addition, the cytopathic effect on Madin-Darby Canine Kidney cells coincubated with N. fowleri diminished in the presence of antibodies against Nf23; nevertheless, the nonpathogenic amoebae did not produce damage to the monolayer cells. These results suggest that the plasma membrane protein Nf23 is probably involved in the virulence of N. fowleri.
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Naegleria fowleri/metabolismo , Naegleria fowleri/patogenicidade , Naegleria/metabolismo , Naegleria/patogenicidade , Proteínas de Protozoários/metabolismo , Virulência/genética , Animais , Encéfalo/metabolismo , Encéfalo/parasitologia , Cães , Expressão Gênica , Células Madin Darby de Rim Canino , Camundongos , Naegleria fowleri/genética , Proteínas de Protozoários/genética , Análise de Sequência de ProteínaRESUMO
INTRODUCTION: The acute kidney injury (AKI) is characterized by a sudden glomerular filtration reduction. Renal or intrinsic causes of AKI include nephrotoxicity induced by exogenous agents like cisplatin, which causes oxidative stress altering the biochemical process and leading to apoptosis. Therefore, this research is aimed at analyzing the embryonic stem cells (ESC) nephroprotective effect in AKI induced by cisplatin, employing genetic, phenotypic, and microspectroscopic techniques. METHODS: Thirty mice were randomly divided into three groups (n = 10): the healthy, isotonic salt solution (ISS), and mouse embryonic stem cells (mESC) groups. The ISS and mESC groups were subjected to AKI using cisplatin; 24 h post-AKI received an intraperitoneal injection of ISS or 1 × 106 mESC, respectively. At days 4 and 8 post-AKI, five mice of each group were sacrificed to analyze the histopathological, genetic (PDK4 and HO-1), protein (p53), and vibrational microspectroscopic changes. RESULTS: Histopathologically, interstitial nephritis and acute tubular necrosis were observed; however, the mESC group showed a more preserved microarchitecture with high cellularity. Additionally, the PDK4 and HO-1 gene expression only increased in the ISS group on day 4 post-AKI. Likewise, p53 was more immunoexpressed at day 8 post-AKI in the ISS group. About biomolecular analysis by microspectroscopy, bands associated with lipids, proteins, and nucleic acids were evidenced. Besides, ratios related to membrane function (protein/lipid), unsaturated lipid content (olefinic/total lipid, olefinic/total CH2, and CH2/CH3), and lipid peroxidation demonstrated oxidative stress induction and lipid peroxidation increase mainly in the ISS group. Finally, the principal component analysis discriminated against each group; nonetheless, some data of the healthy and mESC groups at day 8 were correlated. CONCLUSIONS: The mESC implant diminishes cisplatin nephrotoxicity, once the protective effect in the reduction of lipid peroxidation was demonstrated, reflecting a functional and histological restoration.
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Injúria Renal Aguda/induzido quimicamente , Antineoplásicos/efeitos adversos , Cisplatino/efeitos adversos , Células-Tronco Embrionárias/metabolismo , Peroxidação de Lipídeos/genética , Injúria Renal Aguda/patologia , Animais , Humanos , CamundongosRESUMO
RESUMEN El estudio de la microbiota bacteriana asociada con periodontitis es esencial para desarrollar herramientas de diagnóstico y eficacia en las terapias, por esta razón el objetivo de este estudio fue identificar bacterias asociadas con periodontitis. Nosotros amplificamos el gen 16S rARN por reacción en cadena de polimerasa (PCR) y secuenciamos para investigar y comparar muestras subgingivales obtenidas de individuos sanos y pacientes con periodontitis moderada y severa. Identificamos Stenotrophomonas maltophilia, Porphyromonas gingivalis, Pseudomonas sp., Fusobacterium nucleatum, Haemophilus sp., Aggregatibacter sp. y Prevotella intermedia. P. gingivalis y F. nucleatum se asociaron con ambas periodontitis y Aggregatibacter sp. se asoció con periodontitis severa. Los resultados presentados aquí podrían contribuir en la toma de decisiones clínicas de la periodontitis.
ABSTRACT The study of bacterial microbiota associated with periodontitis is essential to develop effective diagnostic tools and therapies. Hence, the aim of this study was to identify the bacteria associated with periodontitis. We used 16S rRNA gene amplification by polymerase chain reaction (PCR) and sequencing to identify and compare 80 subgingival samples from patients with healthy periodontium and patients with severe and moderate periodontitis. We identified the following bacteria: Stenotrophomonas maltophilia, Porphyromonas gingivalis, Pseudomonas sp., Fusobacterium nucleatum, Haemophilus sp., Aggregatibacter sp., and Prevotella intermedia. P. gingivalis and F. nucleatum were associated with both moderate and severe periodontitis, and Aggregatibacter sp. was associated with severe periodontitis. The results of this research can be useful in clinical decision-making for treatment of patients with periodontitis.
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We evaluate structural damage effects of heat on DNA obtained from the dental pulp of restored premolars. We studied three groups (A, B and C) each group comprised twenty premolars extracted from five patients. Three of the four premolars of each donator were restored with different materials: amalgam, glass ionomer and resin, and one unrestored premolar was used as control. The group A was not exposed to heat, while B and C groups were exposed to 100⯰C and 300⯰C, respectively. The DNA damage was evaluated as percentage of genotyping of 15 Short Tandem Repeats (STRs) and amelogenin and by Fourier-transform infrared spectroscopy (FTIR). The results showed 100% genotyping in all unheated premolars; however, the increase in heat decreased genotyping percentage due to DNA degradation. The amplifications from the premolars restored with glass ionomer and those unrestored were less affected, amplifying by approximately 80% at 300⯰C. FTIR revealed that DNA structural damage occurred in the phosphate region, and changes in ribose were also shown; in addition, we detected presence of ß- three-calcium-phosphate (ß - TCP) due to heat treatment. Moreover, the phosphate region of DNA was a good indicator of DNA integrity related to the ratio of 1230/1085â¯cm-1 in the second derivative (asymmetric/symmetric PO2), which was major in premolars restored with glass ionomer heated at 100⯰C, and this ratio is related to less DNA alterations and better genotyping; however this changes only were detected at 100⯰C, suggesting that dental restoration with this material only protects dental pulp at temperatures below 300⯰C.
Assuntos
Dano ao DNA , DNA/metabolismo , Polpa Dentária , Temperatura Alta/efeitos adversos , Adolescente , Adulto , Amelogenina/metabolismo , Dente Pré-Molar , Fosfatos de Cálcio/metabolismo , Criança , Dano ao DNA/genética , Restauração Dentária Permanente , Feminino , Genótipo , Técnicas de Genotipagem , Humanos , Masculino , Repetições de Microssatélites/genética , Ribose/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , Adulto JovemRESUMO
The aim of this study was to evaluate the association between prostate cancer (PCa) and Human papillomavirus (HPV) infection in the Mexican population. We studied 356 paraffin-embedded tissues from unrelated Mexican men with PCa or benign prostatic hyperplasia (BPH), with the latter serving as control. HPV detection was performed by polymerase chain reaction (PCR) using universal primers, and viral genotypes were detected using sequencing or multiplex PCR. Light microscopy analyses enabled the identification of koilocytes in samples subsequently analyzed for HPV detection by in situ PCR and for p16-INK4A expression by immunohistochemistry. The results showed that high risk- (HR) HPVs were detected in 37/189 (19.6%) PCa specimens compared to 16/167 (9.6%) of BHP specimens (odds ratio 2.3; 95% CI= 1.2 to 4.3; p=0.01). These data suggest HR-HPV may play a role in PCa. HPV 52 and 58 were the most frequent genotypes (33 and 17%, respectively) detected in the population studied. Koilocytes were detected in all in situ PCR-HPV-positive samples, representing a pathognomonic feature of infection, and we observed the overexpression of p16-INK4A in HPV-positive samples compared to HPV-negative samples, indirectly suggesting the presence of HR-HPV E7 oncoprotein. These results suggest that HPV infection plays an important role in prostate cancer development.
RESUMO
Oxygen or nitrogen oxidative species and chemical stress induce the programmed cell death (PCD) of Entamoeba histolytica trophozoites. PCD caused by the aminoglycoside G418 is reduced by incubation with the cysteine protease inhibitor E-64; however, no typical caspases or metacaspases have been detected in this parasite. Calpain, a cysteine protease activated by calcium, has been suggested to be part of a specific PCD pathway in this parasite because the specific calpain inhibitor Z-Leu-Leu-Leu-al diminishes the PCD of trophozoites. Here, we predicted the hypothetical 3D structure of a calpain-like protein of E. histolytica and produced specific antibodies against it. We detected the protein in the cytoplasm and near the nucleus. Its expression gradually increased during incubation with G418, with the highest level after 9 h of treatment. In addition, a specific calpain-like siRNA sequence reduced the cell death rate by 65%. All these results support the hypothesis that the calpain-like protein is one of the proteases involved in the execution phase of PCD in E. histolytica. The hypothetical interactome of the calpain-like protein suggests that it may activate or regulate other proteins that probably participate in PCD, including those with EF-hand domains or other calcium-binding sites.
Assuntos
Apoptose , Calpaína/metabolismo , Entamoeba histolytica/fisiologia , Calpaína/antagonistas & inibidores , Biologia Computacional , Entamoeba histolytica/efeitos dos fármacos , Ativadores de Enzimas/metabolismo , Gentamicinas/metabolismo , Mapas de Interação de ProteínasRESUMO
Genetic factors, such as the mitochondrial DNA (mtDNA) T16189C polymorphism, have been associated with metabolic syndrome (MetS), but this association has not been studied in Mexico to date. The aim of the present study was to determine whether this polymorphism contributes to MetS in the Mexican population. We recruited 100 unrelated volunteer subjects who were divided into 2 groups: with MetS (MetS group) and without MetS (control group). All subjects were genotyped for the mtDNA T16189C polymorphism by polymerase chain reaction and sequencing. The mitochondrial T16189C polymorphism was detected in 24 (24%) of 100 subjects analyzed. The frequency of the mtDNA T16189C polymorphism was higher in the MetS group with 21 (32.3%) of 65 testing positive compared to 3 (8.5%) of 35 in the control group, indicating that this polymorphism is a probable risk factor for MetS in the Mexican population (odds ratio 5.0909, 95% CI 1.3977-18.5424, P = 0.0136). Our results may contribute to early diagnosis of MetS, which is essential for establishing changes in early stages of the disease to avoid further complications and pathologies, thereby preventing the development of type 2 diabetes and cardiovascular disease in Mexico.