Molecular and serological diagnosis of toxoplasmosis: a systematic review and meta-analysis.

Toxoplasmosis is an infection of vast worldwide distribution whose etiologic agent is Toxoplasma gondii. This disease can cause problems ranging from mild symptoms to serious conditions, such as encephalitis, miscarriage and blindness. Therefore, it is of utmost importance to perform a diagnosis with reproducible techniques in order to obtain a good prognosis. The aim of this review was to analyze the efficiency of toxoplasmosis diagnostic techniques based on sensitivity and specificity results. Five research platforms in English language were used (Eric, Elsevier, Google Scholar, PubMed and SciELO), which contained data on the diagnosis of toxoplasmosis. The search and selection were performed for studies published prior to June 2021. The search resulted in the inclusion of 13 articles published from 2005 to 2020. The data revealed the use of different samples in the standardization of techniques such as serum, total blood, colostrum and amniotic fluid. The flow cytometry, lateral flow immunoassay and qPCR techniques showed 100% sensitivity, whereas the ELISA, western blotting, qPCR and RE-LAMP techniques achieved 100% specificity. Significantly, the qPCR and LAMP techniques were more accurate when the likelihood ratio was assessed. The meta-analysis identified that ISAGA and western blotting have low sensitivity values and LIASON, ELFA and ELISA, using a silica bioconjugate, also have low specificity values. It was noted that a wide range of methods have high values of sensitivity and specificity. Therefore, the choice of the method will be based on the conditions and its financial viability.

Molecular and serological diagnosis of toxoplasmosis: a systematic review and meta-analysis

ABSTRACT Toxoplasmosis is an infection of vast worldwide distribution whose etiologic agent is Toxoplasma gondii. This disease can cause problems ranging from mild symptoms to serious conditions, such as encephalitis, miscarriage and blindness. Therefore, it is of utmost importance to perform a diagnosis with reproducible techniques in order to obtain a good prognosis. The aim of this review was to analyze the efficiency of toxoplasmosis diagnostic techniques based on sensitivity and specificity results. Five research platforms in English language were used (Eric, Elsevier, Google Scholar, PubMed and SciELO), which contained data on the diagnosis of toxoplasmosis. The search and selection were performed for studies published prior to June 2021. The search resulted in the inclusion of 13 articles published from 2005 to 2020. The data revealed the use of different samples in the standardization of techniques such as serum, total blood, colostrum and amniotic fluid. The flow cytometry, lateral flow immunoassay and qPCR techniques showed 100% sensitivity, whereas the ELISA, western blotting, qPCR and RE-LAMP techniques achieved 100% specificity. Significantly, the qPCR and LAMP techniques were more accurate when the likelihood ratio was assessed. The meta-analysis identified that ISAGA and western blotting have low sensitivity values and LIASON, ELFA and ELISA, using a silica bioconjugate, also have low specificity values. It was noted that a wide range of methods have high values of sensitivity and specificity. Therefore, the choice of the method will be based on the conditions and its financial viability.

A systematic review of Toxoplasma gondii genotypes in Gallus gallus domesticus worldwide: The focus is Brazil.

Toxoplasma gondii was initially classified in three main lineages related to its virulence: Types I, II, and III. The recombination of genes during sexual cycle in felids gut led to more than 200 genotypes, found in ToxoDB database, using 11 RFLP markers. Free-range chickens are good bioindicators of soil contamination with T. gondii oocysts. In this sense, there are systematic reviews regarding data of genetic characterization of this parasite in felines and ruminants, but not in chickens heretofore, what makes this work necessary. A systematic review of the literature was performed with papers published prior to September 21, 2020. The main inclusion criteria were the presence of T. gondii genotypes, isolated strictly from free-range chickens, in experimental works. Initially, a total of 1,343 studies related to the terms were identified on databases and 30 studies were selected to be systematically reviewed. A total of 561 isolates of T. gondii from 6,356 free-range chickens were analyzed for genotyping, revealing 190 genotypes. ToxoDB #59 and #2 were the most frequent in America, #1 was the most frequent in Africa and three atypical isolates from genotype ToxoDB #9 were found in Asia. There is no data from Europe and Oceania. The majority of studies were Brazilian (16/30). A total of 68 RFLP genotypes were recognized among the 561 isolates' DNAs analyzed from the 30 studies. Some studies showed new genotypes never described before, which reinforces the idea that in some years even more new genotypes will be identified, due to gradual genetic recombination. A large number of undefined genotypes makes it necessary to perform Nested PCR technique when genotyping. Moreover, the lack of data in Continents such as Europe, Asia, and Oceania makes it necessary to perform new isolating and genotyping studies in these places.

Effects of corticosterone treatment and wound healing on reproductive traits of American bullfrogs.

During reproductive season, calling anuran males display high testosterone (T) and episodically high corticosterone (CORT) plasma levels, which are positively associated with higher calling rates and immunocompetence. However, exposure to constant stress stimuli can result in chronically elevated CORT levels, possibly leading to inhibition of reproductive and immune activity. Reproduction and immune responses are energetically expensive, so when an animal is immunologically challenged, a tradeoff might be expressed, with CORT potentially mediating it. Our aim was to test how episodic and chronic CORT treatments, alongside wound healing, would affect reproduction in American bullfrog males (Lithobates catesbeianus). Forty animals were divided in four groups: Episodic CORT (daily transdermic application of CORT), placebo (daily transdermic application of sesame oil), chronic CORT (subcutaneous CORT silastic implants), and sham control (subcutaneous empty silastic implants). One week after treatments began, animals were punctured in the leg with a biopsy needle and the wound was photographed after 45 days to determine wound healing status (WS). Blood samples were collected throughout the experiment to measure CORT and T plasma levels. After animal euthanasia, testes were dissected, fixed, and analyzed histologically to determine spermatogenic activity (germinative cyst [GmC] morphometrics). As expected, the episodic CORT treatment had no effect on T plasma levels or spermatogenic activity. On the other hand, chronic CORT treatment reduced GmC morphometric traits, indicating suppression of reproduction, although T levels were not altered. In addition, animals from sham control and chronic CORT treatments with higher T levels presented higher WS, which indicates an immune-enhancing T effect.

Short-term diuretic withdrawal in stable outpatients with mild heart failure and no fluid retention receiving optimal therapy: a double-blind, multicentre, randomized trial.

AIMS: Although loop diuretics are widely used to treat heart failure (HF), there is scarce contemporary data to guide diuretic adjustments in the outpatient setting. METHODS AND RESULTS: In a prospective, randomized and double-blind protocol, we tested the safety and tolerability of withdrawing low-dose furosemide in stable HF outpatients at 11 HF clinics in Brazil. The trial had two blindly adjudicated co-primary outcomes: (i) symptoms assessment quantified as the area under the curve (AUC) of a dyspnoea score on a visual-analogue scale evaluated at 4 time-points (baseline, Day 15, Day 45, and Day 90) and (ii) the proportion of patients maintained without diuretic reuse during follow-up. We enrolled 188 patients (25% females; 59 ± 13 years old; left ventricular ejection fraction = 32 ± 8%) that were randomized to furosemide withdrawal (n = 95) or maintenance (n = 93). For the first co-primary endpoint, no significant difference in patients' assessment of dyspnoea was observed in the comparison of furosemide withdrawal with continuous administration [median AUC 1875 (interquartile range, IQR 383-3360) and 1541 (IQR 474-3124), respectively; P = 0.94]. For the second co-primary endpoint, 70 patients (75.3%) in the withdrawal group and 77 patients (83.7%) in the maintenance group were free of furosemide reuse during follow-up (odds ratio for additional furosemide use with withdrawal 1.69, 95% confidence interval 0.82-3.49; P = 0.16). Heart failure-related events (hospitalizations, emergency room visits, and deaths) were infrequent and similar between groups (P = 1.0). CONCLUSIONS: Diuretic withdrawal did not result in neither increased self-perception of dyspnoea nor increased need of furosemide reuse. Diuretic discontinuation may deserve consideration in stable outpatients with no signs of fluid retention receiving optimal medical therapy. CLINICALTRIALS.GOV IDENTIFIER: NCT02689180.

DONORS (Donation Network to Optimise Organ Recovery Study): Study protocol to evaluate the implementation of an evidence-based checklist for brain-dead potential organ donor management in intensive care units, a cluster randomised trial.

INTRODUCTION: There is an increasing demand for multi-organ donors for organ transplantation programmes. This study protocol describes the Donation Network to Optimise Organ Recovery Study, a planned cluster randomised controlled trial that aims to evaluate the effectiveness of the implementation of an evidence-based, goal-directed checklist for brain-dead potential organ donor management in intensive care units (ICUs) in reducing the loss of potential donors due to cardiac arrest. METHODS AND ANALYSIS: The study will include ICUs of at least 60 Brazilian sites with an average of ≥10 annual notifications of valid potential organ donors. Hospitals will be randomly assigned (with a 1:1 allocation ratio) to the intervention group, which will involve the implementation of an evidence-based, goal-directed checklist for potential organ donor maintenance, or the control group, which will maintain the usual care practices of the ICU. Team members from all participating ICUs will receive training on how to conduct family interviews for organ donation. The primary outcome will be loss of potential donors due to cardiac arrest. Secondary outcomes will include the number of actual organ donors and the number of organs recovered per actual donor. ETHICS AND DISSEMINATION: The institutional review board (IRB) of the coordinating centre and of each participating site individually approved the study. We requested a waiver of informed consent for the IRB of each site. Study results will be disseminated to the general medical community through publications in peer-reviewed medical journals. TRIAL REGISTRATION NUMBER: NCT03179020; Pre-results.

Lipases obtained from orange wastes: Commercialization potential and biochemical properties of different varieties and fractions.

Brazil is the world's leading orange supplier for juice production purposes. However, the production process generates high amount of wastes, which leads to disposal problems. Orange wastes can be used for lipases production, incorporating the biorefinery concept into juice industries. Thus, the aim of the present study was to investigate the wastes of orange production chain as source of lipases based on different varieties (Pera, Hamlin, Valencia, and Natal), as well as on different fractions of wastes. The mass balance of the juice/wastes (2007-2016 crops) was evaluated, and lipases from different varieties and fraction were biochemically characterized. Overall, the wastes corresponded to approximately 43% of the fruit mass. All the fractions of all varieties showed lipase activity in emulsified olive oil and in p-nitrophenyl substrates. The highest lipase activities were obtained by Natal pulp in emulsified olive oil, Natal frit, and Hamlin peel in p-NPB and Hamlin frit in p-NPL and p-NPP. The bagasse, peel, and frit lipases from the different orange varieties showed optimum pH from 6.0 to 8.0 and optimal temperature from 30 °C to 60 °C. Thus, it is possible concluding that the orange processing for juice production purposes generates a large amount of wastes, which can be destined to profitable purposes as lipases production. Lipases produced by different fractions and varieties are biochemically diverse, enabling the application a wide range of processes. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2734, 2019.

Production of milk peptides with antimicrobial and antioxidant properties through fungal proteases.

Bioactive peptides can provide health benefits due to different mechanisms. The aims of the present study are to produce bioactive peptides from bovine and goat milk subjected to the proteolytic activity of Aspergillus oryzae and Aspergillus flavipes enzymes, as well as to assess their putative antimicrobial and antioxidant activity. Bioactive peptides were successfully generated from proteases of fungi cultivated in solid-state fermentation. The generated peptides were effective against all tested bacteria and fungi. There was antioxidant activity, up to 92.5% DPPH reduction and ORAC stabilization at 52.5 µmol µL-1 of Trolox Equivalent. The generation of milk-specific sequences peptides in the samples was obtained through 2D-PAGE fractioning followed by mass spectrometry (ESI-MS/MS). Based on results in the present study, milk bioactive peptides presenting broad antimicrobial action and antioxidant activity spectra can be cost-effectively produced through solid-state fermentation. The herein addressed approach can be valuable for the pharmaceutical and food industries.

Brazilian fruit processing, wastes as a source of lipase and other biotechnological products: a review.

The global food loss and waste is the most urgent research area in food science to attend the current demand for more sustainable and profitable processes. Along the productive chain about 1/3 of the food is lost or wasted, this number reaches 1/2 for fruit and vegetable production in developing countries. Brazil has been investing in researches aiming to turn its wastes into byproducts, as biomolecules of high value such as lipases. These enzymes are found in a high diversity of plant sources and their researches are covered by promising market growth expectations due to the current demand for biofuels and bio-transformed food. Thus, the aim of the present study is to discuss the potential of wastes generated by the Brazilian fruit processing to become a source of lipases, by the analysis of the most recent studies on fruit lipases, as well as the inclusion of this process in the biorefinery concept. According to this concept, different products can be obtained from the same raw material. Considering the confirmation of the presence of lipases on fruit wastes, the annual fruit production and the percentage of residues, the assessed data showed that wastes from the processing of orange, mango, papaya and palmare promising for lipase obtainment.

Improvement of lipase obtaining system by orange waste-based solid-state fermentation: production, characterization and application.

Lipases are an economic important group of biocatalysts that can be produced by some fungal under solid-state fermentation. Orange wastes are source of lipases and potential substrates for lipases production. This work assessed 19 fugal strains cultivated in Citrus sinensis cv. Hamlin orange wastes (peel, frit and core) for production of lipases in order to generate compounds with antioxidant, antimicrobial and cytotoxic properties. Fifteen of those fungi grew and produced lipases, mainly the Aspergillus brasiliensis [National Institute of Quality Control (INCQS) 40036]/frit system, which showed 99.58 U/g total lipase. The substrate with the highest production of lipase was frit with 26.67 and 78.91 U/g of total lipases produced on average by the 15 microorganisms. Aspergillus niger 01/frit (33.53 U/g) and Aspergillus niger (INCQS 40015)/frit (34.76 U/g) systems showed the highest specificity values in all the herein tested synthetic substrates with 4, 12 and 16 carbons. Analysis of the fatty acid profile of hydrolysis products obtained in the most prominent systems applied to corn and sunflower oils showed: palmitic acid, linoleic acid, oleic acid, and stearic acid. These acids showed antioxidant capacity of up to 58% DPPH (2,2-diphenyl-1-pierylhydrazyl) radical reduction and antibacterial activity against Escherichia coli, Listeria monocytogenes, Pseudomonas aureginosa, Salmonella Enteritidis and Staphylococcus aureus, as well as cytotoxicity to SCC9 cells (squamous cancer cells).

Bioremediation of cooking oil waste using lipases from wastes.

Cooking oil waste leads to well-known environmental impacts and its bioremediation by lipase-based enzymatic activity can minimize the high cytotoxic potential. In addition, they are among the biocatalysts most commercialized worldwide due to the versatility of reactions and substrates. However, although lipases are able to process cooking oil wastes, the products generated from this process do not necessarily become less toxic. Thus, the aim of the current study is to analyze the bioremediation of lipase-catalyzed cooking oil wastes, as well as their effect on the cytotoxicity of both the oil and its waste before and after enzymatic treatment. Thus, assessed the post-frying modification in soybean oil and in its waste, which was caused by hydrolysis reaction catalyzed by commercial and home-made lipases. The presence of lipases in the extracts obtained from orange wastes was identified by zymography. The profile of the fatty acid esters formed after these reactions was detected and quantified through gas chromatography and fatty acids profile compared through multivariate statistical analyses. Finally, the soybean oil and its waste, with and without enzymatic treatment, were assessed for toxicity in cytotoxicity assays conducted in vitro using fibroblast cell culture. The soybean oil wastes treated with core and frit lipases through transesterification reaction were less toxic than the untreated oils, thus confirming that cooking oil wastes can be bioremediated using orange lipases.

Field effect of two commercial preparations of botulinum toxin type A: a prospective, double-blind, randomized clinical trial.

BACKGROUND: The dose equivalence of commonly used commercial preparations of botulinum toxin type A, Dysport (abotulinumtoxinA [ABO] 500 U, Ipsen Biopharm Limited, Wrexham, United Kingdom) and Botox (onabotulinumtoxinA [ONA] 100 U, Allergan, Irvine, CA), remains unclear. OBJECTIVE: We sought to evaluate the field effect for ABO and ONA at dose equivalences of 2.5:1.0 U and 2.0:1.0 U, in both muscular and sweat gland activity. METHODS: In all, 59 female patients with forehead wrinkles were enrolled. Patients were randomized for dose equivalence between ABO and ONA, group A (2.0:1.0 U, ABO:ONA) or group B (2.5:1.0 U, ABO:ONA) administered in the frontalis muscles. Clinical assessment, Minor test, and electromyography evaluations were performed at baseline, 28 days, and 112 days. RESULTS: In group B, the field of anhidrotic effect of ABO showed a greater area and larger horizontal diameter than ONA at 28 and 112 days. At maximum frontalis muscle activity (day 112) patients receiving ABO demonstrated greater improvement based on the Wrinkle Severity Scale. No differences were found in frontalis muscle activity at rest between groups A and B based on results of Wrinkle Severity Scale, electromyography, and interindividual variability data at 28 and 112 days. LIMITATIONS: Currently, there are no objective measurements other than electromyography to evaluate the field effect of botulinum toxin type A in muscles. CONCLUSION: At a dose equivalence of 2.0:1.0 U (ABO:ONA), similar field effects were found for both muscle and sweat gland activity. At a higher dose equivalence of 2.5:1.0 U (ABO:ONA), injections of ABO showed greater area and larger horizontal diameter in field of anhidrotic effect at 28 and 112 days than ONA.

Celluqol® - instrumento de avaliação de qualidade de vida em pacientes com celulite / CELLUQOL® - a quality of life measurement fot patients with cellulite

Introdução: Após a puberdade, a maioria das mulheres desenvolve algum grau de celulite. O aumento do número de consultas dermatológicas relacionadas ao tratamento da celulite demandou compreensão mais ampla de características, desejos e expectativas desses pacientes com relação à qualidade de vida. Objetivo: Elaborar e validar instrumento de avaliação da qualidade de vida para pacientes com celulite.Métodos: O estudo teve duas etapas: a elaboração e a validação dos instrumentos de avaliação da qualidade de vida. Na segunda etapa, dois questionários foram elaborados e aplicados a 100 voluntárias com idade entre 18 e 45 anos, que apresentavam celulite.Resultados: A confiabilidade dos resultados foi verificada por meio de análise fatorial e Teste a de Cronbach para avaliação estatística. Uma análise fatorial exploratória possibilitou testar ahipótese de que todas as questões do questionário resumido mediam um único domínio ou fator (no caso, celulite) e a hipótese de que cada bloco de questões do questionário completo media um único domínio ou fator entre os parâmetros avaliados (modo de vestir, lazer, atividade física, parceiro, sentimentos e mudança de hábitos cotidianos). Conclusões: A análise de validação mostrou que ambos os questionários podem ser usados, com grau similar de eficácia.

Side-by-side comparison of areas with and without cellulite depressions using magnetic resonance imaging.

BACKGROUND: Cellulite is characterized by alterations in the relief of the skin surface. Magnetic resonance imaging (MRI) is recognized as a reliable technique for measuring adipose volume according to body site and for the visualization of the subcutaneous structures. OBJECTIVE: To compare subcutaneous tissue in areas with and without cellulite on the buttocks of same subjects using a noninvasive technique. METHODS AND MATERIALS: Thirty female patients with cellulite on the buttocks underwent MRI. An area with cellulite and another without cellulite on the contralateral buttock were selected. Two soft gelatin capsules of different sizes were used as skin markers to differentiate the areas with and without cellulite. RESULTS: Fibrous septa were visualized in 96.7% of the area with cellulite depressions; most of them were ramified (73.3%) and presented a high-intensity signal on T2 images (70%). All fibrous septa found in the examined areas were perpendicular to the skin surface. The average fibrous septa thickness was 2.18 +/- 0.89 in the area with cellulite and 0.27 +/- 0.64 in the area without cellulite. CONCLUSION: Results of the MRI analysis showed that cellulite depressions on the buttocks were significantly associated with the presence of underlying fibrous septa.

Blind multicenter study of the efficacy and safety of injections of a commercial preparation of botulinum toxin type A reconstituted up to 15 days before injection.

BACKGROUND: It is recommended that attention be given to the shelf life of botulinum toxin type A (BT-A) after its reconstitution. OBJECTIVE: To assess the efficacy and safety of 500 U of BT-A after reconstitution up to 15 days after injection. MATERIAL AND METHODS: BT-A vials were diluted 15 days, 8 days, and 8 hours before injection. One hundred five volunteers were randomized to one of three treatment groups, according to dilution dates. They were evaluated at baseline and 28, 56, 84, and 112 days after treatment. At each visit, the investigator and the volunteer evaluated the motility of the treated area using a 4-point qualitative scale. Five independent specialists, who scored the motility of the treated area on the same scale, blind analyzed photographs taken at each visit. The reconstituted vials of BT-A were stored and analyzed before and after the study. RESULTS: No significant difference was shown between the groups. No evidence of contamination was found in the BT-A vials. CONCLUSION: The results confirm the possibility of injecting 500 U of BT-A up to 15 days after its reconstitution safely and without loss of efficacy.

A randomized pilot study comparing the action halos of two commercial preparations of botulinum toxin type A.

BACKGROUND: The determination of the action halos of botulinum toxin type A aids in targeting specific muscles and/or sweat glands, thereby preventing the occurrence of side effects. OBJECTIVES: The objective of this study was to compare the action halos of two commercial preparations of botulinum toxin type A, Dysport (Ipsen, Slough, UK) and BOTOX (Allergan, Irvine, CA). MATERIAL AND METHODS: Eighteen volunteers received applications of both products into randomized sides of the frontalis muscle. Equivalent doses of 5 U of Dysport and 2 U of BOTOX were injected using the same technique, in the same volume (0.02 mL), and at a controlled depth. Twenty-eight days later, clinical and photographic assessments were performed. RESULTS: All the areas around the injected points were regular, round, or slightly oval and showed similar effects in the muscles and sweat glands. No statistically significant differences were observed between the mean sizes of the diameters of the halos produced by the two products. CONCLUSION: Injections of Dysport and BOTOX at an equivalence ratio of 2.5:1 U, respectively, applied at the same volume and depth, using the same technique resulted in similar action halos with regard to muscular and sweat gland activity. Both products seem to be safe and very predictable.

Botanical extracts used in the treatment of cellulite.

BACKGROUND: Cellulite is defined as skin relief alterations that give the skin an orange peel or mattress appearance. The lesions tend to be asymptomatic and may be considered the anatomic expressions of the structures in the affected area, such as the fat and subcutaneous septa. OBJECTIVE: The present article reviews the most important botanical extracts used as active ingredients in the treatment of cellulite, as well as the steps to obtain these botanicals as raw material and their standardization and quality control, which are important to guarantee their therapeutic action. METHODS: The current literature was reviewed, and we also obtained information from the manufacturers of the products that contained botanicals because of the few publications about this subject. CONCLUSIONS: The reduction in fat deposits through the continuous use of anticellulite products depends on the availability of the active ingredient at the action site, the concentration of the ingredient in the formulation, and the physiochemical characteristics particular to each active ingredient. The botanicals used in topical products must have standardized extracts, which would permit each phytomedicine to have the same effect anywhere in the world. New scientific research is necessary to verify the efficacy and ideal concentrations of such substances.