Preparation and evaluation of a niosomal delivery system containing G. mangostana extract and study of its anti-Acanthamoeba activity.

Garcinia mangostana extract (GME) has severe pharmacokinetic deficiencies and is made up of a variety of bioactive components. GME has proven its anti-Acanthamoeba effectiveness. In this investigation, a GME-loaded niosome was developed to increase its potential therapeutic efficacy. A GME-loaded niosome was prepared by encapsulation in a mixture of span60, cholesterol, and chloroform by the thin film hydration method. The vesicle size, zeta potential, percentage of entrapment efficiency, and stability of GME-loaded niosomes were investigated. The values for GME-loaded niosome size and zeta potential were 404.23 ± 4.59 and -32.03 ± 0.95, respectively. The delivery system enhanced the anti-Acanthamoeba activity, which possessed MIC values of 0.25-4 mg mL-1. In addition, the niosomal formulation decreased the toxicity of GME by 16 times. GME-loaded niosome must be stored at 4 °C, as the quantity of remaining GME encapsulated is greater at this temperature than at room temperature. SEM revealed the damage to the cell membrane caused by trophozoites and cysts, which led to dead cells. In light of the above, it was found that GME-loaded niosomes had better anti-Acanthamoeba activity. The study suggested that GME-loaded niosomes could be used as an alternative to Acanthamoeba's therapeutic effects.

Cognitive-affective manifestations since premanifest phases of Spinocerebellar Ataxia Type 3/Machado-Joseph Disease.

BACKGROUND: Cognitive deficits were related to Spinocerebellar Ataxia type 3/Machado-Joseph Disease (SCA3/MJD), but the Cerebellar Cognitive Affective Syndrome (CCAS) needs further investigation in this disorder. We aimed to characterize cognitive-affective deficits in manifest and premanifest SCA3/MJD carriers. METHODS: Subjects at 50% risk, manifest carriers and unrelated controls were evaluated in-person or in virtual settings with CCAS Scale (CCAS-S), Stroop Color-Word Test (SCWT), Trail-Making Test (TMT), and Reading the Mind in the Eyes Test (RMET). Scale for Assessment and Rating of Ataxia (SARA) >2.5 or Friedreich Ataxia Rating Scale/Activities of Daily Living (FARS-adl) >4 divided carriers into manifest and premanifest. Time after onset or time left to gait ataxia onset (TimeToAfterOnset) were estimated. Differences between groups and correlations with TimeToAfterOnset, SARA and FARS-adl were checked. RESULTS: After random selection to balance groups, 23 manifest and 35 premanifest carriers, and 58 controls were included. CCAS-S, semantic fluency, phonemic fluency, category switching, affect, SCWT, and RMET showed significant differences between manifest carriers and controls; premanifest carriers mostly displayed intermediate values between controls and manifest carriers. These variables correlated with TimeToAfterOnset and SARA scores of the carriers. Correlations with SARA were stronger in the pre-ataxic group. CCAS-S had the strongest correlations with time and SARA. DISCUSSION: Cognitive-affective deficits in SCA3/MJD involve executive function, language, affect, and social cognition, which seem to be altered prior to the ataxia onset, and correlate with markers of motor progression. CCAS-S was the most promising biomarker and should be evaluated in longitudinal studies.

Food insecurity and nutritional status among older adults: a systematic review.

CONTEXT: Food insecurity (FI), characterized by difficulty or inability to access adequate food, has become a public health problem. OBJECTIVE: To analyze studies relating FI with nutritional status (NS) among older adults and the associated factors. DATA SEARCH: Articles published up to June 2020 were investigated in 5 databases: PubMed, Embase, Scopus, LILACS, and Web of Science. The search, selection, extraction, and quality evaluation were carried out by 2 reviewers. DATA EXTRACTION: The authors identified characteristics of the studies and the main data regarding the relationship of interest. RESULTS: Twenty-two studies were included in the review and their characteristics are summarized and presented using narrative synthesis. In 10 studies (45.4%), a relationship was observed between FI and malnutrition; in another 6 (27.3%), a relationship was observed between FI and being overweight. CONCLUSION: A relationship was identified between FI, especially severe forms, and malnutrition, as well as between FI, especially mild forms, and people being overweight. Thus, FI among older adults relates to a 2-fold burden of nutritional outcomes, depending on the level. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration no. CRD42020185086.

Probiotics and Amelioration of Rheumatoid Arthritis: Significant Roles of Lactobacillus casei and Lactobacillus acidophilus.

Rheumatoid arthritis is a chronic autoimmune disorder that can lead to disability conditions with swollen joints, pain, stiffness, cartilage degradation, and osteoporosis. Genetic, epigenetic, sex-specific factors, smoking, air pollution, food, oral hygiene, periodontitis, Prevotella, and imbalance in the gastrointestinal microbiota are possible sources of the initiation or progression of rheumatoid arthritis, although the detailed mechanisms still need to be elucidated. Probiotics containing Lactobacillus spp. are commonly used as alleviating agents or food supplements to manage diarrhea, dysentery, develop immunity, and maintain general health. The mechanism of action of Lactobacillus spp. against rheumatoid arthritis is still not clearly known to date. In this narrative review, we recapitulate the findings of recent studies to understand the overall pathogenesis of rheumatoid arthritis and the roles of probiotics, particularly L. casei or L. acidophilus, in the management of rheumatoid arthritis in clinical and preclinical studies.

COVID-19 in a state of Brazilian Northeast: Prevalence and associated factors in people with flu-like syndrome.

AIMS AND OBJECTIVES: To estimate the prevalence and associated factors of COVID-19 in people with flu-like syndrome in Ceará, Brazil. BACKGROUND: COVID-19 is an infectious disease that has led to a worldwide public health emergency. More than 30,000 cases were confirmed in Brazil, especially in the States of São Paulo, Rio de Janeiro and Ceará. The capital of the Ceará State is the one with the highest incidence of COVID-19 in Brazil. Estimating the prevalence of the disease and its associated factors is important to offer adequate health care. DESIGN: A cross-sectional study with secondary data of people notified with flu-like syndrome and COVID-19 test results. METHODS: 19,967 cases of flu-like syndrome were analysed according to the result of the COVID-19 test. Predictive variables were as follows: age range, sex, women in puerperium, presence or absence of cardiovascular diseases, diabetes, haematological illness, immunodeficiencies, neurological diseases, obesity, renal diseases and Down syndrome. Robust Poisson regression models estimated the prevalence ratios of COVID-19. The research was reported via STROBE guidelines for cross-sectional studies. RESULTS: The prevalence of COVID-19 in the population was 10.37%. In the final model, the following variables were associated with COVID-19: aged people, male sex, cardiovascular diseases and diabetes. CONCLUSION: Among the flu-like syndrome cases, COVID-19 prevalence was high. In the Ceará State, clinical factors such as aged people, male sex, cardiovascular diseases and diabetes can enhance the prevalence of COVID-19 by up to 2.57 times. RELEVANCE TO CLINICAL PRACTICE: The identification of factors that are associated with the enhanced prevalence of COVID-19 facilitates early diagnosis, and adequate and prompt treatment. This knowledge may avoid an unfavourable prognosis of the disease.

Multiorgan histopathological changes in the juvenile seabream Sparus aurata as a biomarker for zinc oxide particles toxicity.

Zinc oxide nanoparticles are widely used in some domains (cosmetics, pharmaceuticals optical devices, and agricultural field) due to their physical, optical, and antimicrobial properties. However, the release of ZnO-NPs into the environment may affect organisms like fish with potential consequences for human health. Histological approaches of the acute effects of these materials on fish are scarce; thus, the present study aimed to investigate the potential toxic effects of acute exposure to ZnO particles in marine environments, by assessing histological changes in the gills, liver, spleen, and muscle of gilthead seabream (Sparus aurata) juveniles. Thus, fish were exposed for 96 h, via water, to 1 mg L-1 of ionic zinc and zinc oxide particles (1.1, 1.2, and 1.4 µm of size). Histological examination revealed gills as the most affected organ, followed by liver, muscle, and spleen. In the gills, histopathological changes included hyperplasia of epithelial cells, fusion of the secondary lamellae, and lifting of the lamellar epithelium with edema. In the liver, lipid vacuolation of several degrees, necrosis of hepatic and pancreatic tissues, blood congestion in sinusoids and hepatoportal vessels, presence of cellular infiltrate, and melano-macrophages diffusion was found. Muscle showed degeneration, atrophy, thickening and necrosis of muscle fibers with edema between them, and presence of melano-macrophages in the muscle layer. Spleen was the less damaged organ, displaying congested blood, white pulp increase/rupture, and bigger and darker melano-macrophage aggregates in the splenic stroma. These results underline that the size of particles plays a determinant role in their potential pernicious effects. A short-term exposure caused major histopathological changes in relevant organs of S. aurata juveniles, possibly affecting their function.

Fungal Diversity of Human Gut Microbiota Among Eutrophic, Overweight, and Obese Individuals Based on Aerobic Culture-Dependent Approach.

Fungi have a complex role in the intestinal tract, influencing health and disease, with dysbiosis contributing to obesity. Our objectives were to investigate fungal diversity in human gut microbiota among eutrophic, overweight, and obese. Epidemiological and nutritional information were collected from adult individuals, as well as stool samples processed for selective fungi isolation and identification by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (yeasts) or microculture (filamentous fungi). Further 18S rDNA sequencing was performed to confirm identification. The mean count of fungi was 241 CFU/g of feces. Differences in the population level of the filamentous fungi were observed within eutrophic and obese groups. Overall, 34 genera were identified. The predominant phylum was Ascomycota with 20 different genera, followed by Basidiomycota and Zygomycota. As for Ascomycota, the most prevalent species were Paecilomyces sp., Penicillium sp., Candida sp., Aspergillus sp., Fonsecaea sp., and Geotrichum sp. (76.39, 65.28, 59.72, 58.33, 12.50, and 9.72%, respectively). As for Basidiomycota, Trichosporon sp. and Rhodotorula sp. were the most prevalent (30.56 and 15.28%, respectively), and for Zygomycota, Rhizopus sp. and Mucor sp. were the most numerous (15.28 and 9.72%, respectively). As expected there is a mycobiota shift towards obesity, with slightly higher diversity associated to eutrophic individuals. This mycobiota shift seems also to be related to the nutritional behavior of the individuals, as observed that the macronutrients intake may be positively related to the different fungi occurrences. Other studies are needed to better understand relationships between mycobiota and obesity, which could be used in future obesity treatments.

Toward establishing a morphological and ultrastructural characterization of proembryogenic masses and early somatic embryos of Araucaria angustifolia (Bert.) O. Kuntze.

Somatic embryogenesis is a morphogenetic route useful for the study of embryonic development, as well as the large-scale propagation of endangered species, such as the Brazilian pine (Araucaria angustifolia). In the present study, we investigated the morphological and ultrastructural organization of A. angustifolia somatic embryo development by means of optical and electron microscopy. The proembryogenic stage was characterized by the proliferation of proembryogenic masses (PEMs), which are cellular aggregates composed of embryogenic cells (ECs) attached to suspensor-like cells (SCs). PEMs proliferate through three developmental stages, PEM I, II, and III, by changes in the number of ECs and SCs. PEM III-to-early somatic embryo (SE) transition was characterized by compact clusters of ECs growing out of PEM III, albeit still connected to it by SCs. Early SEs showed a dense globular embryonic mass (EM) and suspensor region (SR) connected by embryonic tube cells (TCs). By comparison, early somatic and zygotic embryos showed similar morphology. ECs are round with a large nucleus, nucleoli, and many cytoplasmic organelles. In contrast, TCs and SCs are elongated and vacuolated with cellular dismantling which is associated with programmed cell death of SCs. Abundant starch grains were observed in the TCs and SCs, while proteins were more abundant in the ECs. Based on the results of this study, a fate map of SE development in A. angustifolia is, for the first time, proposed. Additionally, this study shows the cell biology of SE development of this primitive gymnosperm which may be useful in evolutionary studies in this area.

Genetic diagnosis in recently transfused patients.

Analysis of recently transfused patients is usually postponed to avoid spurious results because of contamination with donor's cells. However, little is known about the extent of this influence in routine molecular diagnostic tests. To elucidate this question, we tested a mix of blood samples from 2 α-1-antitrypsin-deficient patients diagnosed as Pi*Z homozygous with 1 normal donor at 1:1, 1:10, 1:20, and 1:30 proportions. Human identification panel and Pi*Z allele detection were used to establish the detection limit of a blood mixture. Mixtures of 1:1 and 1:10 were easily detected with both techniques, whereas for 1:30, it was necessary to change the equipment settings to identify the mixture. Moreover, the heterozygous pattern observed for the mixtures on Pi*Z genotyping was weaker at this level of mixture. We further evaluated the degree of mixture detectable in 20 transfused patients who received 1 blood unit (concentrate of irradiated or nonirradiated red blood cells) using the human identification panel. Two days after the transfusion, the presence of the donor's markers was not detected, suggesting that after this time point the levels of admixture are below 1:30. The methods applied in the present study showed adequate sensitivity to identify alleles of the so-called "smaller population" of cells up to 3%, approximately. The same result was obtained in a "diagnostic situation," in which the blood mixture was submitted to a PCR-RFLP protocol to detect a mutation.

Morphological, compositional and ultrastructural changes in the Scrobicularia plana shell in response to environmental mercury--an indelible fingerprint of metal exposure?

The study aimed to assess morphological, structural and compositional alterations in Scrobicularia plana nacre environmentally exposed to mercury in order to seek out the possibility of the assessed alterations as a monitoring tool to handle complexity and interactions of metals in the environment involving a non-invasive methodology. Bivalves were collected from a mercury contaminated site (Laranjo basin - Ria de Aveiro, Portugal) and a reference site in the same aquatic system. The combination of scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDS) technique depicted a sheet like morphology of bivalve nacre collected from the reference site. Moreover, EDS plot exhibited the presence of potassium, oxygen, calcium, and carbon elements. Shells collected from the contaminated area depicted lamellar patches like structures with particle like morphology composition. SEM images corresponding to the elemental analysis by EDS plot clearly denoted the presence of mercury. SEM images from the other locations of the contaminated shells depicted large surface area, a broken or ruptured symmetry of organic matrix as well as crack-like gaps. The influence of environmental mercury affecting the surface morphology of S. plana nacre showed dimple like morphology (as proved by transmission electron microscopy, TEM). The possible explanation may be the replacement of calcium elements with other elements or alloys from the nacre composite collected from contaminated region. Therefore, the nacre fingerprint may be useful as innovative knowledge and applicable tool aiming at risk reduction from noxious mercury present in the environment. Overall results suggested the use of shell as an indelible fingerprint of metal exposure.

Mode of sexual differentiation and its influence on the relative sensitivity of the fathead minnow and zebrafish in the fish sexual development test.

Exogenous treatment of fish with natural sex hormones and their mimics has been shown to influence gonadal differentiation resulting in biased phenotypic sex-ratios. This has lead to the development of the Fish Sexual Development Test (FSDT) as a method for the detection of endocrine active chemicals. Proposed test organisms include the medaka, zebrafish (ZF) and stickleback, although the guideline also allows for inclusion of species such as the fathead minnow (FHM), provided the test duration allows for sufficient sexual differentiation. However, although the processes underlying sexual differentiation are known to differ for each of these species, it is not known how, or if, these differences would influence the results of the FSDT. In the experiments reported here, responses of the ZF and FHM to prochloraz, a sterol biosynthesis inhibitor and androgen antagonist, were characterized and compared. Exposure to 320 µg/L of prochloraz, from embryo until 60 (ZF) or 95-125 (FHM) days post hatch inhibited somatic growth of both species, but while a negative impact on ZF larval survival was observed (LOEC 32 µg/L) there was no evidence for an effect on FHM larval survival. Prochloraz influenced sexual differentiation in both species by decreasing the proportion of females (LOEC 100 µg/L (ZF), 320 µg/L (FHM)) and delaying completion of sexual differentiation; manifest as an increased incidence of ovotestis in the ZF (LOEC 100 µg/L) and as an increased number of fish with undifferentiated gonads in the FHM (LOEC 320 µg/L). However, while exposure to 320 µg/L prochloraz delayed maturation of the differentiated FHM testis, there was no such effect in the ZF. These results demonstrate that the different strategy of sexual differentiation in the ZF and FHM influences the profile of responses of their gonads to the masculinising effects of prochloraz, but does not affect their overall sensitivity.

Simultaneous effects of nicotine, acrolein, and acetaldehyde on osteogenic-induced bone marrow cells cultured on plasma-sprayed titanium implants.

PURPOSE: To evaluate the potential interaction/contribution of inductive and deleterious effects of tobacco compounds on human osteoblastic cells cultured on plasma-sprayed titanium implants exposed to combinations of nicotine, acrolein, and acetaldehyde. Cell response was assessed as proliferation and function. MATERIALS AND METHODS: Titanium implants, seeded with human bone marrow-derived cells (first subculture), were cultured in osteogenic-inducing conditions for 28 days in the absence (control) and in the presence of tobacco compounds to assess (1) the dose-dependent profile of acrolein (0.01 to 0.12 mmol/L) and acetaldehyde (0.1 to 6 mmol/L) and (2) the effect of the simultaneous exposure to combinations of nicotine, acrolein, and acetaldehyde. In later experiments, seeded implants were exposed to two different concentrations of nicotine (1.2 mmol/L, known to have inductive effects on cell behavior, and 2.4 mmol/L, reported to elicit deleterious effects on cell behavior) with acrolein, acetaldehyde, or both, at a concentration that inhibits 50% (IC50). RESULTS: Acrolein and acetaldehyde caused dose-dependent inhibitory effects at levels similar to and greater than 0.03 and 0.1 mmol/L, respectively; IC50 regarding cell viability/proliferation and alkaline phosphatase was 0.06 mmol/L for acrolein and 0.3 mmol/L for acetaldehyde. Matrix mineralization was prevented at levels higher than 0.03 mmol/L acrolein and 0.1 mmol/L acetaldehyde. Exposure to a combination of nicotine 1.2 mmol/L with acrolein (0.06 mmol/L), acetaldehyde (0.3 mmol/L), or both resulted in a cell behavior intermediate to that observed in nicotine-treated cultures (induced cell response) and aldehyde-treated cultures (deleterious cell response). On the other hand, exposure to nicotine 2.4 mmol/L with acrolein (0.06 mmol/L), acetaldehyde (0.3 mmol/L), or both caused cumulative cytotoxic responses. CONCLUSION: Results suggest that interactions of tobacco compounds on osteoblasts might contribute to the overall effects of tobacco use on implant osseointegration and long-time survival.

Lunularia cruciata, a potential in vitro host for Glomus proliferum and G. intraradices.

A study was conducted to define culture conditions for in vitro growth arbuscular mycorrhizal fungi (AMF) with liverworts as hosts. Lunularia cruciata (L.) Dumortier ex. Lindberg developed in vitro monoxenic mycothalli with both Glomus proliferum Dalpé & Declerck (MUCL 41827) and Glomus intraradices Schenck & Smith (MUCL 43204). AMF inoculated plants were co-cultured in plastic Petri dishes with semi-solidified medium supplemented with sucrose and grown under filtered light. Mycothalli of L. cruciata produced external hyphae and spores in quantities equivalent to those obtained with Ri T-DNA transformed root systems.

S100B and NSE serum concentrations in Machado Joseph disease.

BACKGROUND: NSE and S100B are considered as neuronal and glial peripheral markers of central nervous system pathologies, respectively. We evaluated the potential use of S100B and NSE serum concentrations as peripheral markers of symptomatic patients with Machado Joseph disease (MJD). METHODS: We measured S100B and NSE peripheral concentrations of 22 MJD patients and compared with healthy subjects concentrations. The correlations of both markers with CAG repeat size, age of onset, disease duration, and the scores of the Extended Disability Status Scale of Kurtzke, Unified Parkinson's Disease Rating Scale, and the Montgomery-Asberg depression rating scale were also assessed. RESULTS: S100B serum concentrations between control and MJD subjects were not statistically different, whereas NSE serum concentrations were higher in MJD patients than in control subjects (p=0.00001). S100B presented a moderate correlation with disease duration and depression score, whereas NSE presented a moderate correlation with depression score and a good negative correlation with EDSS score. CONCLUSIONS: Symptomatic MJD patients present increased concentrations of NSE and normal concentrations of S100B in blood.