Entamoeba histolytica: In Silico and In Vitro Oligomerization of EhHSTF5 Enhances Its Binding to the HSE of the EhPgp5 Gene Promoter.

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.

Feasibility in the use of the UV-Visible region for the characterization of glucose in deionized water using Arduino 1 / Viabilidad en el uso de la región UV-Visible para la caracterización de glucosa en agua desionizada mediante Arduino 1

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.

Dynamic response antibodies SARS-CoV-2 human saliva studied using two-dimensional correlation (2DCOS) infrared spectral analysis coupled with receiver operation characteristics analysis.

COVID-19 has affected the entire world due to the rapid spread of SARS-CoV-2, mainly through airborne particles from saliva, which, being easily obtained, help monitor the progression of the disease. Fourier transform infrared (FTIR) spectra combined with chemometric analysis could increase the diagnostic efficiency of the disease. However, two-dimensional correlation spectroscopy (2DCOS) is superior to conventional spectra as it helps to resolve the minute overlapped peaks. In this work, we aimed to use 2DCOS and receiver operating characteristic (ROC) analyses to compare the immune response in saliva associated with COVID-19, which could be important in biomedical diagnosis. FTIR spectra of human saliva samples from male (575) and female (366) patients ranging from 20 to 85 ± 2 years of age were used for the study. Age groups were segregated as G1 (20-40 ± 2 years), G2 (45-60 ± 2 years), and G3 (65-85 ± 2 years). The results of the 2DCOS analysis showed biomolecular changes in response to SARS-CoV-2. 2DCOS analyses of the male G1 + (1579,1644) and -(1531,1598) cross peaks evidenced changes such as amide I > IgG. Female G1 cross peaks -(1504,1645), (1504,1545) and -(1391,1645) resulted in amide I > IgG > IgM. The asynchronous spectra in 1300-900 cm-1 of the G2 male group showed that IgM is more important in diagnosing infections than IgA. Female G2 asynchronous spectra -(1027,1242) and + (1068,1176) showed that IgA > IgM is produced against SARS-CoV-2. The G3 male group evidenced antibody changes in IgG > IgM. The absence of IgM in the female G3 population diagnoses a specifically targeted immunoglobulin associated with sex. Moreover, ROC analysis showed sensitivity (85-89 % men; 81-88 % women) and specificity (90-93 % men; 78-92 % women) for the samples studied. The general classification performance (F1 score) of the studied samples is high for the male (88-91 %) and female (80-90 %) populations. This high PPV (positive predictive value) and NPV (negative predictive value) verify our segregation of COVID-19 positive and negative sample groups. Therefore, 2DCOS with ROC analysis using FTIR spectra have the potential for a non-invasive approach to monitoring COVID-19.

Riluzole, a Derivative of Benzothiazole as a Potential Anti-Amoebic Agent against Entamoeba histolytica.

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.

Oxidative stress and genotoxicity in oral epithelial cells from subjects undergoing orthodontic treatment with fixed appliances.

OBJECTIVES: The objective of this work was to evaluate the impact of fixed orthodontic appliances on oxidative stress (OS) and genotoxicity from oral epithelial cells. MATERIALS AND METHODS: Samples of oral epithelial cells were obtained from fifty-one healthy voluntary subjects who had an indication for orthodontic treatment. The samples were obtained before treatment and after 6 and 9 months of treatment. OS was evaluated by quantitating 8-hydroxy-2'deoxyguanosine (8-OHdG) and by performing relative gene expression with antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). DNA degradation and instability were evaluated by multiplex polymerase chain reaction (PCR) and fragment analysis for human identification. RESULTS: The quantitation results showed that 8-OHdG increased during treatment, although this increase was not statistically significant. SOD increased by 2.5- and 2.6-fold after 6 and 9 months of treatment, respectively. CAT increased by threefold after 6 months of treatment, while after 9 months of treatment, the expression level decreased to a level similar to that before treatment. DNA degradation was found in 8% and 12% of DNA samples after 6 and 9 months of treatment, respectively, while DNA instability was detected in only 2% and 8% of DNA samples after 6 and 9 months of treatment, respectively. CONCLUSIONS: The results showed that OS and genotoxicity slightly changed after treatment with a fixed orthodontic appliance; in addition, a biological adaptation response to the treatment may occur after 6 months. CLINICAL RELEVANCE: OS and genotoxicity in the buccal cavity are risk factors for oral and systemic diseases. This risk may be reduced through antioxidant supplementation, by using thermoplastic materials, or by reducing the orthodontic treatment time.

Prolonged Computer Use by Office Workers Induces Ocular Symptoms Associated With Tear Film Alterations and Overexpression of Mucin 5 AC and Catalase.

OBJECTIVE: The aim of the study was to evaluate office workers for symptoms of computer vision syndrome (CVS) and alterations in the tear film relate to the hours of daily computer use. METHODS: Sixty-seven volunteers were divided into 2 groups: 2 to 6 and 7 to 12 hours of daily computer use. Computer vision syndrome symptoms, tear film stability by tear film break-up time test, and composition of mucin 5 AC, catalase, and IL-6 was assessed by relative gene expression of conjunctival impression cytology samples were examined. RESULTS: All participants exhibited moderate symptoms of CVS, whereas 90% showed reduced tear film stability. For the 7- to 12-hour (vs 2- to 6-hour) group, these effects were more pronounced and overexpression of mucin 5 AC and catalase was detected. CONCLUSIONS: Prolonged computer use induced an overexpression of mucin 5 AC and catalase and instability of the tear film, associated with ocular symptoms.

Evaluación del polimorfismo Pro12Ala del gen PPARƴ en mujeres con diabetes mellitus gestacional / Evaluation of the Pro12Ala polymorphism of the PPARƴ gene in women with gestational diabetes mellitus

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.

Comparison of the Immune Response in Vaccinated People Positive and Negative to SARS-CoV-2 Employing FTIR Spectroscopy.

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-ɑ), 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.

Evaluation of Cytochrome P4502E1 mRNA Expression and Its Effects in Antioxidant Defenses, and Cell Toxicity in Printing Workers.

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.

High-Risk HPV with Multiple Infections Promotes CYP2E1, Lipoperoxidation and Pro-Inflammatory Cytokines in Semen of Asymptomatic Infertile Men.

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.

Short tandem repeat (STR) instability in the oral mucosa of patients submitted to fixed orthodontic therapy: a limitation of STR profile quality for human identification.

The purpose of this study was to evaluate changes in short tandem repeat (STR) profile quality before and after fixed orthodontic therapy. Samples of oral epithelial cells were obtained from 28 volunteers who had an indication for orthodontic treatment. The samples were collected before and three months after starting orthodontic treatment with fixed appliances. DNA extraction and integrity were evaluated by electrophoresis, and STR profiles were obtained by polymerase chain reaction amplification and STR typing via capillary electrophoresis. DNA electrophoresis showed a higher proportion (7/28, 25%) of DNA degradation in the samples collected after fixed orthodontic treatment compared to those obtained before starting orthodontic therapy (3/28, 11%), however, changes in DNA were not significant (p=0.289). In concordance all STR profiles showed complete genotyping; however, imbalances in the size of heterozygotes and in the signal were detected in 25% of STR profiles after orthodontic therapy. Moreover, STR instability was demonstrated by an increase in stutter bands detected in 60% of the DNA profiles after treatment and a spurious allele of the D195433 marker was found in one sample after treatment. The STR profiles of samples obtained from the oral cavity with orthodontic appliances should be interpreted with caution. STR instability increases the incidence of artifacts that could compromise the quality of the results of tests performed in forensic DNA laboratories.

Correlation of the DNA Concentration of Human Samples to Electrical Bioimpedance Measurements: A Pilot Study.

It is necessary to evaluate the total deoxyribonucleic acid (DNA) concentration in gene expression assays. The existing techniques require equipment that is expensive for many labs in developing countries. Portable and inexpensive equipment is needed for easy and economical DNA quantification. Electrical bioimpedance spectroscopy (EBiS) is a non-invasive and inexpensive technique for examining the electrical properties of biological materials. The aim of this study was to explore a potential correlation between the measurement of total DNA extracted from human samples by UV-Vis spectrophotometry and EBiS. Hence, after quantifying the total DNA extracted from each sample by UV-Vis spectroscopy, EBiS was recorded and a possible correlation between the two measurements was analyzed. Considering the bioimpedance phase parameter at 5.24 MHz, a significant correlation was found with total DNA, especially when the concentration was below 100 ng/µL (Spearman coefficient = 0.82, p<0.005). Additional experiments are warranted to confirm these findings.

Proinflammatory and Oxidative Stress States Induced by Human Papillomavirus and Chlamydia trachomatis Coinfection Affect Sperm Quality in Asymptomatic Infertile Men.

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.

Seminal pro-inflammatory cytokines and pH are affected by Chlamydia infection in asymptomatic patients with teratozoospermia.

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.

Evaluation of Individualized Homeopathic Treatment in Patients With Irritable Bowel Syndrome: A Pilot Study.

BACKGROUND: Some studies have suggested the efficacy of homeopathic treatment for irritable bowel syndrome (IBS). OBJECTIVE: The aim of this pilot study was to evaluate the efficacy of individualized homeopathic treatment in patients with IBS. METHODS: The study was carried out at the National Homeopathic Hospital of the Secretary of Health, Mexico City, Mexico and included 41 patients: 3 men and 38 women, mean age 54 ± 14.89 years, diagnosed with IBS as defined by the Rome IV Diagnostic criteria. Single individualized medicine was prescribed for each patient, taking into account all presenting symptoms, clinical history, and personality via repertorization using RADAR Homeopathic Software (archibel, Isnes, Belgium). The homeopathic drugs were used at fifty-millesimal (LM) potency per the Mexican Homeopathic Pharmacopoeia starting with 0/1 and increasing every month (0/2, 0/3, 0/6). Severity scales were applied at the beginning of treatment and every month for 4 months of treatment. The evaluation was based on comparing symptom severity scales during treatment. RESULTS: The results demonstrated that 100% of patients showed some improvement and 63% showed major improvement or were cured. The study showed a significant decrease in severity of symptom scores 3 months after treatment, with the pain score showing a decrease 1 month after treatment. The results highlight the importance of individualized medicine regimens using LM potency, although the early decrease in pain observed could also be due to the fact that Lycopodium clavatum and Nux vomica were the main homeopathic medicine prescribed, and these medicines contain many types of alkaloids, which have shown significant analgesic effects on pain caused by physical and chemical stimulation. CONCLUSION: This pilot study suggests that individualized homeopathic treatment using LM potencies benefits patients with IBS.

(-)-Epicatechin protects from amebic liver abscess development in hamster.

In this work the effect of (-)-epicatechin on the development of amebic liver abscess in hamsters was evaluated. (-)-epicatechin is a flavonoid present in plants that possesses various biological properties, including its activity against some protozoal parasites; however its antiamebic activity in a living model had not been evaluated. Syrian golden hamsters were intrahepatically inoculated with 1x106E. histolytica trophozoites, three days after inoculation they received nine intraperitoneal doses of (-)-epicatechin (10 mg/100 g) every 48 h. Animals without treatments and treated with metronidazole were included as controls. Macroscopic characteristics of the hepatic abscess, histopathological analysis of the tissue and the levels of inflammatory cytokines were determined. (-)-epicatechin produced a decrease in liver abscess progression being observed only 9.49% of damage compared to 84% shown by untreated animals. During treatment with (-)-epicatechin hepatic tissue showed signs of liver repair and absence of amoebae. Additionally, (-)-epicatechin produced a modulating effect on inflammatory cytokines TNF-α, IL-1ß and IL-10. All these events observed in animals treated with (-)-epicatechin could contribute to the elimination of trophozoites and liver healing.

Case reports: Electroacupuncture decreases the climacteric symptoms by calcitonin gene-related peptide modulation.

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.

Pulmonary Alterations Among Workers in a Dental Prosthesis Laboratory: Exploring High Dust Concentrations and Novel Findings of Bacterial Genera in the Workplace to Achieve Improved Control.

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.

The MPO system participates actively in the formation of an oxidative environment produced by neutrophils and activates the antioxidant mechanism of Naegleria fowleri.

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.

A comparative study of the membrane proteins from Naegleria species: A 23-kDa protein participates in the virulence of Naegleria fowleri.

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.