Detection and Molecular Identification of Eight Candida Species in Clinical Samples by Simplex PCR.

Systemic candidiasis is a frequent opportunistic mycosis that can be life-threatening. Its main etiological agent is Candida albicans; however, the isolation of non-albicans Candida species has been increasing. Some of these species exhibit greater resistance to antifungals, so the rapid and specific identification of yeasts is crucial for a timely diagnosis and optimal treatment of patients. Multiple molecular assays have been developed, based mainly on polymerase chain reaction (PCR), showing high specificity and sensitivity to detect and identify Candida spp. Nevertheless, its application in diagnosis has been limited due to specialized infrastructure or methodological complexity. The objective of this study was to develop a PCR assay that detects and identifies some of the most common pathogenic Candida species and evaluate their diagnostic utility in blood samples and bronchial lavage. A pair of oligonucleotides was designed, CandF and CandR, based on sequence analysis of the 18S-ITS1-5.8S-ITS2-28S region of the rDNA of Candida spp., deposited in GenBank. The designed oligonucleotides identified C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, C. krusei/Pichia kudriazevii, C. guilliermondii/Meyerozyma guilliermondii, C. lusitaniae/Clavispora lusitaniae, and C. dubliniensis using simplex PCR based on the amplicon size, showing a detection limit of 10 pg/µL of DNA or 103 yeasts/mL. Based on cultures as the gold standard, it was determined that the sensitivity (73.9%), specificity (96.3%), and the positive (94.4%) and negative (81.2%) predictive values of the PCR assay with the designed oligonucleotides justify their reliable use in diagnosis.

Does the Fetus Limit Antibiotic Treatment in Pregnant Patients with COVID-19?

During pregnancy, there is a state of immune tolerance that predisposes them to viral infection, causing maternal-fetal vulnerability to the adverse effects of COVID-19. Bacterial coinfections significantly increase the mortality rate for COVID-19. However, it is known that all drugs, including antibiotics, will enter the fetal circulation in a variable degree despite the role of the placenta as a protective barrier and can cause teratogenesis or other malformations depending on the timing of exposure to the drug. Also, it is important to consider the impact of the indiscriminate use of antibiotics during pregnancy can alter both the maternal and fetal-neonatal microbiota, generating future repercussions in both. In the present study, the literature for treating bacterial coinfections in pregnant women with COVID-19 is reviewed. In turn, we present the findings in 50 pregnant women hospitalized diagnosed with SARS-CoV-2 without previous treatment with antibiotics; moreover, a bacteriological culture of sample types was performed. Seven pregnant women had coinfection with Staphylococcus haemolyticus, Staphylococcus epidermidis, Streptococcus agalactiae, Escherichia coli ESBL +, biotype 1 and 2, Acinetobacter jahnsonii, Enterococcus faecium, and Clostridium difficile. When performing the antibiogram, resistance to multiple drugs was found, such as macrolides, aminoglycosides, sulfa, dihydrofolate reductase inhibitors, beta-lactams, etc. The purpose of this study was to generate more scientific evidence on the better use of antibiotics in these patients. Because of this, it is important to perform an antibiogram to prevent abuse of empirical antibiotic treatment with antibiotics in pregnant women diagnosed with SARS-CoV-2.

Candida glabrata Antifungal Resistance and Virulence Factors, a Perfect Pathogenic Combination.

In recent years, a progressive increase in the incidence of invasive fungal infections (IFIs) caused by Candida glabrata has been observed. The objective of this literature review was to study the epidemiology, drug resistance, and virulence factors associated with the C. glabrata complex. For this purpose, a systematic review (January 2001-February 2021) was conducted on the PubMed, Scielo, and Cochrane search engines with the following terms: "C. glabrata complex (C. glabrata sensu stricto, C. nivariensis, C. bracarensis)" associated with "pathogenicity" or "epidemiology" or "antibiotics resistance" or "virulence factors" with language restrictions of English and Spanish. One hundred and ninety-nine articles were found during the search. Various mechanisms of drug resistance to azoles, polyenes, and echinocandins were found for the C. glabrata complex, depending on the geographical region. Among the mechanisms found are the overexpression of drug transporters, gene mutations that alter thermotolerance, the generation of hypervirulence due to increased adhesion factors, and modifications in vital enzymes that produce cell wall proteins that prevent the activity of drugs designed for its inhibition. In addition, it was observed that the C. glabrata complex has virulence factors such as the production of proteases, phospholipases, and hemolysins, and the formation of biofilms that allows the complex to evade the host immune response and generate fungal resistance. Because of this, the C. glabrata complex possesses a perfect pathogenetic combination for the invasion of the immunocompromised host.

Antifungal Resistance in Clinical Isolates of Candida glabrata in Ibero-America.

In different regions worldwide, there exists an intra-and inter-regional variability in the rates of resistance to antifungal agents in Candida glabrata, highlighting the importance of understanding the epidemiology and antifungal susceptibility profiles of C. glabrata in each region. However, in some regions, such as Ibero-America, limited data are available in this context. Therefore, in the present study, a systematic review was conducted to determine the antifungal resistance in C. glabrata in Ibero-America over the last five years. A literature search for articles published between January 2015 and December 2020 was conducted without language restrictions, using the PubMed, Embase, Cochrane Library, and LILACS databases. The search terms that were used were "Candida glabrata" AND "antifungal resistance" AND "Country", and 22 publications were retrieved from different countries. The use of azoles (fluconazole, itraconazole, voriconazole, posaconazole, isavuconazole, ketoconazole, and miconazole) varied between 4.0% and 100%, and that of echinocandins (micafungin, caspofungin, and anidulafungin) between 1.1% and 10.0%. The limited information on this subject in the region of Ibero-America emphasizes the need to identify the pathogens at the species level and perform antifungal susceptibility tests that may lead to the appropriate use of these drugs and the optimal doses in order to avoid the development of antifungal resistance or multi-resistance.

Efecto del manejo de la obesidad clase I con metformina sobre actividad de metaloproteinasas en pacientes con periodontitis crónica / Effect of the management of class I obesity with metformin on metalloproteinase activity in patients with chronic periodontitis

Introducción: el principal problema de salud pública en México es la obesidad y sus enfermedades asociadas, incluyendo las bucales. Objetivo: evaluar el efecto del tratamiento con metformina en pacientes obesos de clase I sobre la actividad de las metaloproteinasas presentes en el periodonto con periodontitis crónica. Métodos: se realizó un estudio clínico con 68 pacientes mujeres con obesidad de clase I y enfermedad periodontal. Se dividieron en 4 grupos; a 2 de ellos, además del tratamiento periodontal, se les administro metformina de 850 mg al día durante seis semanas. Se tomaron 2 muestras por paciente de tejido periodontal antes y después de cada tratamiento y se midió el índice de masa corporal (IMC), el índice de placa dentobacteriana y de inflamación. Mediante zimografía en gel de acrilamida se midió la actividad de las metaloproteinasas en la muestra de tejido recolectada. Los datos obtenidos fueron analizados mediante estadística descriptiva t de student para muestras relacionadas y se realizó ANOVA de una vía considerando p < 0,01 como estadísticamente significativa. Resultados: en el grupo de pacientes a las que se les administro metformina al final del tratamiento se observó una disminución del índice de masa corporal, del grado de inflamación y menor actividad de metaloproteinasas respecto al grupo control (65% frente a 25%; p < 0,01). Conclusiones: el tratamiento con metformina en pacientes con obesidad de clase I y enfermedad periodontal disminuye el IMC, mejora los síntomas de la periodontitis crónica y disminuye la actividad de las metaloproteinasas 1, 3, 8 y V presentes en el periodonto de estos pacientes

Introduction: in Mexico the main problem in public health is obesity and other diseases that are associated whit this condition, including oral health. Objective: to evaluate the effect of metformin treatment in patients with class I obese on the activity of metalloproteinases present in periodontium with chronic periodontitis. Methods: a clinical study was conducted in 68 patients with class I obesity and periodontal disease. They were divided into 4 groups. 2 of them, in addition to the periodontal treatment, were administered metformin 850 mg per day for six weeks; 2 samples were taken per patient of periodontal tissue before and after each treatment, body mass index, plaque index and inflammation were measured. Acrylamide gel zymography was used to measure the activity of metalloproteinases in the sample of tissue collected. The data obtained were analyzed by descriptive statistics, student t for related samples and one-way ANOVA was performed considering p < 0.01 as statistically significant. Results: in the group of patients who were administered metformin at the end of the treatment, there was a decrease in the body mass index, the degree of inflammation and lower metalloproteinase activity, compared with the control group (65% vs 25%; < 0.01). Conclusions: treatment with metformin in patients with obesity class I and periodontal disease decreases BMI, improves the symptoms of chronic periodontitis and decreases the activity of metalloproteinases 1, 3, 8, V present in periodontium of these patients

[Effect of the management of class I obesity with metformin on metalloproteinase activity in patients with chronic periodontitis]. / Efecto del manejo de la obesidad clase I con metformina sobre actividad de metaloproteinasas en pacientes con periodontitis crónica.

INTRODUCTION: Introduction: in Mexico the main problem in public health is obesity and other diseases that are associated whit this condition, including oral health. Objective: to evaluate the effect of metformin treatment in patients with class I obese on the activity of metalloproteinases present in periodontium with chronic periodontitis. Methods: a clinical study was conducted in 68 patients with class I obesity and periodontal disease. They were divided into 4 groups. 2 of them, in addition to the periodontal treatment, were administered metformin 850 mg per day for six weeks; 2 samples were taken per patient of periodontal tissue before and after each treatment, body mass index, plaque index and inflammation were measured. Acrylamide gel zymography was used to measure the activity of metalloproteinases in the sample of tissue collected. The data obtained were analyzed by descriptive statistics, student t for related samples and one-way ANOVA was performed considering p < 0.01 as statistically significant. Results: in the group of patients who were administered metformin at the end of the treatment, there was a decrease in the body mass index, the degree of inflammation and lower metalloproteinase activity, compared with the control group (65% vs 25%; < 0.01). Conclusions: treatment with metformin in patients with obesity class I and periodontal disease decreases BMI, improves the symptoms of chronic periodontitis and decreases the activity of metalloproteinases 1, 3, 8, V present in periodontium of these patients.

INTRODUCCIÓN: Introducción: el principal problema de salud pública en México es la obesidad y sus enfermedades asociadas, incluyendo las bucales. Objetivo: evaluar el efecto del tratamiento con metformina en pacientes obesos de clase I sobre la actividad de las metaloproteinasas presentes en el periodonto con periodontitis crónica. Métodos: se realizó un estudio clínico con 68 pacientes mujeres con obesidad de clase I y enfermedad periodontal. Se dividieron en 4 grupos; a 2 de ellos, además del tratamiento periodontal, se les administro metformina de 850 mg al día durante seis semanas. Se tomaron 2 muestras por paciente de tejido periodontal antes y después de cada tratamiento y se midió el índice de masa corporal (IMC), el índice de placa dentobacteriana y de inflamación. Mediante zimografía en gel de acrilamida se midió la actividad de las metaloproteinasas en la muestra de tejido recolectada. Los datos obtenidos fueron analizados mediante estadística descriptiva t de student para muestras relacionadas y se realizó ANOVA de una vía considerando p < 0,01 como estadísticamente significativa. Resultados: en el grupo de pacientes a las que se les administro metformina al final del tratamiento se observó una disminución del índice de masa corporal, del grado de inflamación y menor actividad de metaloproteinasas respecto al grupo control (65% frente a 25%; p < 0,01). Conclusiones: el tratamiento con metformina en pacientes con obesidad de clase I y enfermedad periodontal disminuye el IMC, mejora los síntomas de la periodontitis crónica y disminuye la actividad de las metaloproteinasas 1, 3, 8 y V presentes en el periodonto de estos pacientes.

Calcarea Carbonica Ostrearum para la obesidad en adolescentes mexicanos / Calcarea Carbonica Ostrearum for mexican adolescent obesity

Antecedentes: la obesidad es un problema de salud pública en México que va en aumento en niños y adolescentes, incrementando el riesgo de enfermedades cardiovasculares y metabólicas a temprana edad. Tradicionalmente, en la práctica clínica homeopática, Calcarea carbonica ostrearum se ha utilizado para la obesidad; sin embargo, no existen estudios de calidad metodológica donde se demuestre sueficacia. El objetivo del estudio fue demostrar la eficacia de Calcarea carbonica ostrearum vs placebo para disminuir la obesidad en adolescentes. Material y métodos: se realizó un ensayo clínico controlado, aleatorizado, doble ciego, de 15 semanas de duración en el Hospital General de México (Ciudad de México) de febrero a agosto de 2010, en el que se incluyó a 25 pacientes adolescentesobesos de entre 14 y 18 años de edad. Se les administró Calcarea carbonica ostrearum 30CH, o placebo, junto con cambios en la dieta y ejercicio. Las variables medidas fueron: peso, índice de masa corporal, perímetro abdominal y porcentaje de grasa corporal. Resultados: se encontraron diferencias estadísticamente significativas entre Calcarea carbonica ostrearum 30CH y placebo en el porcentaje de grasa corporal (Calcarea carbonica ostrearum 2.9±1.1 vs placebo 1.3±1.5; diferencia=1.63, p<0.05). En las demás variables (peso, índice de masa corporal y perímetro abdominal) no se encontraron diferencias estadísticamente significativas entre grupos después de 15 semanas de tratamiento. Conclusión: Calcarea carbonica ostrearum 30CH puede ser efectiva para reducir el porcentaje de grasa corporal en adolescentes obesos. Es necesario que se realicen ensayos clínicos controlados con tamaños de muestra más grandes para corroborar estos resultados. (AU)

Background: obesity is a health problem in Mexico. The prevalence of juvenile and child obesity is raising and is strongly related to cardiovascular and metabolic risk factors. Traditionally, in daily homeopathic practice, Calcarea carbonica ostrearum is prescribed for obesity. Nevertheless there is a lack of well-designed studies to prove its efficacy. The objetive of this study was to investigate the efficacy of Calcarea carbonica ostrearum 30CH vs placebo for reducing obesity in adolescents. Methods/design: a randomized, double-blind, placebo-controlled two-arm trial with a 15 weeks follow-up study was conducted in the Hospital General de Mexico in Mexico City. Twenty-five adolescents between 14 and 18 years were included. The outcomes were: weight, body mass index, abdominal circumference and percentage of body fat. Results: percentage of body fat was the only outcome with statistically significant difference between Calcarea carbonica ostrearum 30CH and placebo (Calcarea carbonica ostrearum 2.9±1.1 vs placebo 1.3±1.5, difference=1.63, p<0.05). Neither weight, nor body mass index, nor abdominal circumference were different from placebo after 15 weeks of treatment. Conclusion: Calcarea carbonica ostrearum 30CH could be effective for reducing the percentage of body fat in obese adolescents. Further studies with larger sample sizes are needed to confirm these results. (AU)

Voice parameters and videonasolaryngoscopy in children with vocal nodules: a longitudinal study, before and after voice therapy.

BACKGROUND: Vocal Nodules (VN) are a functional voice disorder associated with voice misuse and abuse in children. There are few reports addressing vocal parameters in children with VN, especially after a period of vocal rehabilitation. OBJECTIVE: The purpose of this study is to describe measurements of vocal parameters including Fundamental Frequency (FF), Shimmer (S), and Jitter (J), videonasolaryngoscopy examination and clinical perceptual assessment, before and after voice therapy in children with VN. Voice therapy was provided using visual support through Speech-Viewer software. MATERIALS AND METHODS: Twenty patients with VN were studied. An acoustical analysis of voice was performed and compared with data from subjects from a control group matched by age and gender. Also, clinical perceptual assessment of voice and videonasolaryngoscopy were performed to all patients with VN. After a period of voice therapy, provided with visual support using Speech Viewer-III (SV-III-IBM) software, new acoustical analyses, perceptual assessments and videonasolaryngoscopies were performed. RESULTS: Before the onset of voice therapy, there was a significant difference (p<0.05) in mean FF, S and J, between the patients with VN and subjects from the control group. After the voice therapy period, a significant improvement (p<0.05) was found in all acoustic voice parameters. Moreover, perceptual voice analysis demonstrated improvement in all cases. Finally, videonasolaryngoscopy demonstrated that vocal nodules were no longer discernible on the vocal folds in any of the cases. CONCLUSIONS: SV-III software seems to be a safe and reliable method for providing voice therapy in children with VN. Acoustic voice parameters, perceptual data and videonasolaryngoscopy were significantly improved after the speech therapy period was completed.

Effect of caveolin-1 scaffolding peptide and 17beta-estradiol on intracellular Ca2+ kinetics evoked by angiotensin II in human vascular smooth muscle cells.

Caveolae are identifiable plasma membrane invaginations. The main structural proteins of caveolae are the caveolins. There are three caveolins expressed in mammals, designated Cav-1, Cav-2, and Cav-3. It has been postulated that Cav-1 acts as a scaffold protein for signaling proteins; these include ion channels, enzymes, and other ligand receptors like membrane-associated estrogen receptor (ER)alpha or ERbeta. Caveolae-associated membrane proteins are involved in regulating some of the rapid estrogenic effects of 17beta-estradiol. One important system related to the activity of ERalpha and caveolae is the renin-angiotensin system. Angiotensin II (ANG II) has numerous actions in vascular smooth muscle, including modulation of vasomotor tone, cell growth, apoptosis, phosphatidylinositol 3-kinase (PI3K)/Akt activation, and others. Many proteins associated with caveolae are in close relation with the scaffolding domain of Cav-1 (82-101 amino acid residues). It has been proposed that this peptide may acts as a kinase inhibitor. Therefore, to explore the ability of Cav-1 scaffolding peptide (CSP-1) to regulate ANG II function and analyze the relationship between ERalpha and ANG II type 1 and 2 (AT(1) and AT(2)) receptors, we decided to study the effects of CSP-1 on ANG II-induced intracellular Ca(2+) kinetics and the effect of 17beta-estradiol on this modulation using human smooth muscle cells in culture, intracellular Ca(2+) concentration measurements, immuno- and double-immunocytochemistry confocal analysis of receptor expression, immunoblot analysis, and immunocoprecipitation assays to demonstrate coexpression. We hypothesized that CSP-1 inhibits ANG II-mediated increases in intracellular Ca(2+) concentrations by interfering with intracellular signaling including the PI3K/Akt pathway. We also hypothesize that AT(2) receptors associate with Cav-1. Our results show that there is a close association of AT(1), AT(2), and ERalpha with Cav-1 in human arterial smooth muscle cells in culture. CSP-1 inhibits ANG II-induced intracellular signaling.