Cinnamyl Schiff bases: synthesis, cytotoxic effects and antifungal activity of clinical interest.

The aim of this study was to synthesize and investigate the in vitro antifungal properties of 23 cinnamyl Schiff bases. In addition, cytotoxic effects of such cinnamyl Schiff bases against human lung, kidney or red blood cells were also checked. The compounds were synthesized in a single-step, 2 min of reaction under microwave irradiation produced up to 97% yield. Six of the 23 cinnamyl Schiff bases possessed antifungal activities against strains of Candida, Aspergillus, Fonsecaea and, particularly, Cryptococcus species. Indeed, cinnamyl Schiff bases 1 and 23 exhibited minimum inhibitory concentration (MIC) values more than twofold lower than fluconazole (FCZ) against all the Cryptococcus neoformans strains (MIC = 1·33, 1·4 and 5·2 µg ml-1 , respectively) and Cryptococcus gattii strains (MIC = 5·3, 2·8 and 9·2 µg ml-1 , respectively) (12 strains of each species) while cinnamyl Schiff base 11 was as potent as FCZ against all strains from both Cryptococcus species. No significant cytotoxic effects were observed for Schiff bases against human lung, kidney or red blood cells, all presenting selective indexes higher than 10. In conclusion, this study revealed cinnamyl Schiff bases, especially 1 and 23, as new lead anticryptococcal agents for the discovery of novel antifungal drugs. SIGNIFICANCE AND IMPACT OF THE STUDY: The occurrence and severity of fungal infections have increased in recent decades due to resistance to available antifungal drugs and the appearance of new emerging pathogens. Thus, the search for new antifungal agents is mandatory. From a series of 23 cinnamyl Schiff bases, two compounds (1 and 23) were interrogated as new anticryptococcal agents without significant cytotoxicity against human lung, kidney or red blood cells. In turns, these new Schiff bases are lead compounds for the discovery of novel antifungal drugs.

Curcumin enhances the activity of fluconazole against Cryptococcus gattii-induced cryptococcosis infection in mice.

AIM: The aim of this study was to investigate the in vitro and in vivo activities of pure curcumin, as well as its combination with fluconazole, against Cryptococcus gattii. METHODS AND RESULTS: The minimal inhibitory concentrations (MIC) of curcumin and its interactions with fluconazole against C. gattii were assessed in vitro using standard methods. This same combination was used to treat C. gattii-induced cryptococcosis in mice. The behavioural and functional assessment of the mice during treatment was also performed. The average MIC for curcumin was 19·8 µg ml(-1) . Its combination with fluconazole resulted in FICΣ (fractional inhibitory concentration index) values between 0·79 and 2·29. Curcumin (alone or combined with fluconazole) significantly reduced pulmonary damage and fungal burden in the brain. No colonies were found in the brain following combination treatment, which was also confirmed by the improved behaviour of mice. CONCLUSIONS: The combination therapy with curcumin and fluconazole was the most effective among the treatments tested, as in addition to reducing the fungal burden and damage on lung tissues, it was able to eliminate the fungal burden in the brain, enhancing the survival of mice. SIGNIFICANCE AND IMPACT OF THE STUDY: This study points to the possibility of using curcumin in combination with fluconazole as a clinical treatment of cryptococcosis.

Independent Association of Muscular Strength and Carotid Intima-Media Thickness in Children.

The aim of this cross-sectional study was to examine the influence of muscular strength on carotid intima-media thickness (cIMT) in children, controlling for the effect of cardiorespiratory fitness (CRF) and central adiposity and to examine if differences among muscular strength tertiles translate to physiological differences. We assessed cIMT of the common carotid artery in 366 children between 11-12 years of age (191 girls). Measures included cIMT assessed with high-resolution ultrasonography, a maximal handgrip strength test, body fat mass and lean mass from DXA and CRF determined using a maximal cycle ergometer test. Association between muscular strength and cIMT adjusted for CRF and central adiposity, as measured by trunk fat, was tested with multiple linear regression analysis. Differences in risk factors among muscular strength groups were tested with ANOVA. The Muscular Strength Index (MSI) was inversely associated with cIMT independently of CRF and central adiposity (p<0.05). The low MSI group had the highest values of cIMT, waist circumference and systolic blood pressure and the lowest CRF (p<0.05). There was an inverse and independent association between muscular strength and cIMT. Low muscular strength was associated with higher levels of cardiovascular disease risk factors in children.

Increases in intracellular water explain strength and power improvements over a season.

Changes in body components occur over a season, but their impact on performance is still unclear. We aimed to analyze the relationship between changes in leg strength and jump performance with body composition over a season in highly trained athletes. Measures from the beginning to the main competitive periods of a season were obtained in 40 male and 23 female basketball, handball and volleyball players (20±5 years) for fat (FM) and fat-free mass (FFM) estimated by DXA. Total body water (TBW) and extracellular water (ECW) were assessed by deuterium and bromide dilution, respectively, and intracellular water (ICW) was calculated as TBW minus ECW. Maximal strength was determined by the leg press, while jumping height was assessed with squat (SJ) and countermovement (CMJ) jumps. Significant improvements in strength (12.5±20.8%) and jumping height (SJ:8.3±13.9%; CMJ:6.3±8.5%) were found. FFM, TBW and ECW significantly increased (3.0±2.7%; 1.7±5.5%; 3.0±8.6%, respectively), while %FM decreased (-4.5±9.1%) and no changes were observed in ICW (1.2±9.7%). Among body composition changes only ICW was associated with performance even adjusted for gender, age, season length and sport (strength: ß=71.209, p=0.012; SJ: ß=0.311, p=0.049; CMJ: ß=0.366, p=0.018). Body composition, strength and jumping height improved over a season and ICW was the main predictor of performance in national level players.

Association of basketball season with body composition in elite junior players.

AIM: Body composition changes among elite athletes may influence competitive performance. This study aimed to characterize the body composition changes at the molecular, cellular, tissue, and whole-body level of analysis in elite junior basketball players during the course of a season. METHODS: Twelve males and 11 females (16 to 17 years) were evaluated. Dual-energy X-ray absorptiometry (DXA) was used to assess bone mineral (Mo) and lean-soft tissue (LST). Total-body water (TBW) and extracellular water (ECW) were assessed using isotope dilution techniques, and extracellular (ECF) and intracellular fluids (ICF) were calculated. Fat mass (FM) and fat-free mass (FFM) were assessed with a four-compartment model. Body cell mass was calculated (LST - (ECF + ECS)). Skeletal muscle (SM) was estimated using appendicular LST (ALST) as: (1.19 x ALST) - 1.65. At the whole-body level, weight, sum of 7 skinfolds, and muscle circumferences (Mc) were measured. The handgrip and the countermovement jump tests were used for performance assessment. RESULTS: Males increased FFM (4.4±2.3%), TBW (3.5±4.6%), SM (4.5±2.3%), and arm (3.4±2.7%) and thigh (3.8±3%) Mc. Females increased SM (5.9±4.6%) and arm (3.6±3.8%) and thigh (4±5.2%) Mc and decreased ICF (-9.7±13.6%). FFM components differed from the established values based on cadaver analysis. Both genders increased their performance and associations were found between changes in molecular and whole-body components with performance. CONCLUSION: In conclusion the season was associated with an improved body composition profile in males and few changes in females.

Influence of different media, incubation times, and temperatures for determining the MICs of seven antifungal agents against Paracoccidioides brasiliensis by microdilution.

MIC assays with Paracoccidioides brasiliensis, the etiological agent of paracoccidioidomycosis, had been conducted with variable protocols, employing both macrodilution and microdilution tests and including differences in inoculum preparation, media used, incubation periods, and temperatures. Twenty-one clinical and environmental isolates of Paracoccidioides were tested using amphotericin B, itraconazole, ketoconazole, fluconazole, sulfamethoxazole, sulfamethoxazole-trimethoprim, and terbinafine, according to the National Committee for Clinical Laboratory Standards (National Committee for Clinical Laboratory Standards, document M27-A2, 2002), with modifications such as three medium formulations (RPMI 1640 medium, McVeigh and Morton [MVM] medium, and modified Mueller-Hinton [MMH] medium), two incubation temperatures (room temperature [25 to 28 °C] and 37 °C), and three incubation periods (7, 10, and 15 days). The antifungal activities were also classified as fungicidal or fungistatic. The best results were obtained after 15 days of incubation, which was chosen as the standard incubation time. The MICs for most individual isolates grown for the same length of time at the same temperature varied with the different media used (P < 0.05). Of the isolates, 81% showed transition from the yeast to the mycelial form in RPMI 1640 medium at 37 °C, independent of the presence of antifungals. MMH medium appears to be a suitable medium for susceptibility testing of antifungal drugs with P. brasiliensis, except for sulfamethoxazole and the combination of sulfamethoxazole-trimethoprim, for which the MVM medium yielded better results. The incubation temperature influenced the MICs, with, in general, higher MICs at 25 °C (mycelial form) than at 37 °C (P < 0.05). Based on our results, we tentatively propose a microdilution assay protocol for susceptibility testing of antifungal drugs against Paracoccidioides.

Lactate and heart rate variability threshold during resistance exercise in the young and elderly.

This purpose of this study was to: 1) determine the intensity corresponding to anaerobic threshold (AT) during a discontinuous resistance exercise protocol in healthy young and elderly subjects by analyzing heart rate variability (HRV) and blood lactate (BL) and 2) investigate the effect of aging on these variables. A total of 28 individuals, 14 young and 14 elderly healthy men underwent one-repetition maximum (1RM) testing to determine maximum load on the leg press. Discontinuous resistance exercise testing was initiated at 10% of the 1RM with subsequent increases of 10%. The load corresponding to AT was approximately 30% 1RM in both groups. The determination of AT by HRV was associated with BL responses (p<0.01). While HRV indexes decreased with increasing of loads in both groups, the elderly had lower values at loads below AT (p<0.05). Additionally, BL increased sharply after the load corresponding to AT in both groups, although elderly subjects showed the lowest values (p<0.05). In conclusion, HRV is an effective tool for determining AT, which was approximately 30% 1RM under the testing procedures included in the present study. Furthermore, there was a marked change in autonomic function, with gradual vagal withdrawal followed by sympathetic activation. These responses were lower in elderly subjects.

Outbreak of Neisseria meningitidis C in workers at a large food-processing plant in Brazil: challenges of controlling disease spread to the larger community.

SUMMARYAn outbreak of meningococcal disease (MD) with severe morbidity and mortality was investigated in midwestern Brazil in order to identify control measures. A MD case was defined as isolation of Neisseria meningitidis, or detection of polysaccharide antigen in a sterile site, or presence of clinical purpura fulminans, or an epidemiological link with a laboratory-confirmed case-patient, between June and August 2008. In 8 out of 16 MD cases studied, serogroup C ST103 complex was identified. Five (31%) cases had neurological findings and five (31%) died. The attack rate was 12 cases/100 000 town residents and 60 cases/100 000 employees in a large local food-processing plant. We conducted a matched case-control study of eight primary laboratory-confirmed cases (1:4). Factors associated with illness in single variable analysis were work at the processing plant [matched odds ratio (mOR) 22, 95% confidence interval (CI) 2·3-207·7, P<0·01], and residing <1 year in Rio Verde (mOR 7, 95% CI 1·11-43·9, P<0·02). Mass vaccination (>10 000 plant employees) stopped propagation in the plant, but not in the larger community.

Fungal and bacterial coinfections increase mortality of severely ill COVID-19 patients.

BACKGROUND: SARS-CoV-2 predisposes patients to secondary infections; however, a better understanding of the impact of coinfections on the outcome of hospitalized COVID-19 patients is still necessary. AIM: To analyse death risk due to coinfections in COVID-19 patients. METHODS: The odds of death of 212 severely ill COVID-19 patients were evaluated, with detailed focus on the risks for each pathogen, site of infection, comorbidities and length of hospitalization. FINDINGS: The mortality rate was 50.47%. Fungal and/or bacterial isolation occurred in 89 patients, of whom 83.14% died. Coinfected patients stayed hospitalized longer and had an increased odds of dying (odds ratio (OR): 13.45; R2 = 0.31). The risk of death was increased by bacterial (OR: 11.28) and fungal (OR: 5.97) coinfections, with increased levels of creatinine, leucocytes, urea and C-reactive protein. Coinfections increased the risk of death if patients suffered from cardiovascular disease (OR: 11.53), diabetes (OR: 6.00) or obesity (OR: 5.60) in comparison with patients with these comorbidities but without pathogen isolation. The increased risk of death was detected for coagulase-negative Staphylococcus (OR: 25.39), Candida non-albicans (OR: 11.12), S. aureus (OR: 10.72), Acinetobacter spp. (OR: 6.88), Pseudomonas spp. (OR: 4.77), and C. albicans (OR: 3.97). The high-risk sites of infection were blood, tracheal aspirate, and urine. Patients with coinfection undergoing invasive mechanical ventilation were 3.8 times more likely to die than those without positive cultures. CONCLUSION: Severe COVID-19 patients with secondary coinfections required longer hospitalization and had higher risk of death. The early diagnosis of coinfections is essential to identify high-risk patients and to determine the right interventions to reduce mortality.

Correlation of the in vitro antifungal drug susceptibility with the in vivo activity of fluconazole in a murine model of cerebral infection caused by Cryptococcus gattii.

Forty Cryptococcus gattii strains were submitted to antifungal susceptibility testing with fluconazole, itraconazole, amphotericin B and terbinafine. The minimum inhibitory concentration (MIC) ranges were 0.5-64.0 for fluconazole, <0.015-0.25 for itraconazole, 0.015-0.5 for amphotericin B and 0.062-2.0 for terbinafine. A bioassay for the quantitation of fluconazole in murine brain tissue was developed. Swiss mice received daily injections of the antifungal, and their brains were withdrawn at different times over the 14-day study period. The drug concentrations varied from 12.98 to 44.60 µg/mL. This assay was used to evaluate the therapy with fluconazole in a model of infection caused by C. gattii. Swiss mice were infected intracranially and treated with fluconazole for 7, 10 or 14 days. The treatment reduced the fungal burden, but an increase in fungal growth was observed on day 14. The MIC for fluconazole against sequential isolates was 16 µg/mL, except for the isolates obtained from animals treated for 14 days (MIC = 64 µg/mL). The quantitation of cytokines revealed a predominance of IFN-γ and IL-12 in the non-treated group and elevation of IL-4 and IL-10 in the treated group. Our data revealed the possibility of acquired resistance during the antifungal drug therapy.

Social, environmental and microbiologic aspects of endemic mycoses in Brazil.

Many factors that lead to host immunosuppression are clearly known to predispose the host to fungal diseases, significantly influencing the occurrence of mycoses. However, little or nothing has been discussed regarding social or economic factors that can influence the occurrence of diseases caused by fungi. In this minireview, we discuss several factors that may affect the occurrence of mycoses in Brazil, a continentally extended country that is marked by large climatic variations and severe socioeconomic distortions that may limit access to health services for the population.

Genomic and epidemiological monitoring of yellow fever virus transmission potential.

The yellow fever virus (YFV) epidemic in Brazil is the largest in decades. The recent discovery of YFV in Brazilian Aedes species mosquitos highlights a need to monitor the risk of reestablishment of urban YFV transmission in the Americas. We use a suite of epidemiological, spatial, and genomic approaches to characterize YFV transmission. We show that the age and sex distribution of human cases is characteristic of sylvatic transmission. Analysis of YFV cases combined with genomes generated locally reveals an early phase of sylvatic YFV transmission and spatial expansion toward previously YFV-free areas, followed by a rise in viral spillover to humans in late 2016. Our results establish a framework for monitoring YFV transmission in real time that will contribute to a global strategy to eliminate future YFV epidemics.

In vitro cytotoxicity activity on several cancer cell lines of acridone alkaloids and N-phenylethyl-benzamide derivatives from Swinglea glutinosa (Bl.) Merr.

The methanol extract from the stems and fruits of Swinglea glutinosa (Rutaceae) afforded 11 known acridone alkaloids and three N-phenylethyl-benzamide derivatives, glycocitrine-IV, 1,3,5-trihydroxy-4-methoxy-10-methyl-2,8-bis(3-methylbut-2-enyl)acridin-9(10H)-one, 1,3,5- trihydroxy-2,8-bis(3-methylbut-2-enyl)-10-methyl-9-acridone, citbrasine, citrusinine-II, citrusinine-I, 5-dihydroxyacronycine, pyranofoline, 3,4-dihydro-3,5,8-trihydroxy-6-methoxy-2,2,7-trimethyl-2H-pyrano[2,3-a]acridin-12(7H)-one, 2,3-dihydro-4,9-dihydroxy-2-(2-hydroxy-propan-2-yl)-11-methoxy-10-methylfuro[3,2-b]acridin-5(10H)-one, bis-5-hydroxyacronycine, N-(2-{4-[(3,7-dimethylocta-2,6-dien-1-yl)oxy]phenyl}ethyl)benzamide, N-(2-{4-[(3,7-dimethyl-4-acethyl-octa-2,6-dien-1-yl)oxy]phenyl}ethyl)benzamide, and severine acetate. All compounds isolated were examined for their activity against three cancer cell lines: human lung carcinoma (COR-L23), human breast adenocarcinoma (MCF7), human melanoma (C32), and normal human fetal lung cell line, MRC-5. The acridones tested exhibited weak cytotoxicity but the amides showed moderate nonselective cytotoxic activity.

Changes in phase angle and body composition induced by resistance training in older women.

BACKGROUND: Resistance training (RT) has selective effects on body composition that may counteract the deleterious effects of aging. Changes in phase angle (PhA) may serve to monitor the influence of RT in older people. OBJECTIVES: To describe the effect of RT in training, detraining and retraining on body composition, including PhA in older women. SUBJECTS/METHODS: Thirty-three older women (⩾60 years old) participated. The RT program was carried out over 12 weeks for each stage (training, detraining and retraining). Whole-body fat-free mass and fat mass (FM) and appendicular lean soft tissue (ALST) measurements were carried out using a dual energy X-ray absorptiometry. Bioimpedance spectroscopy was used to estimate total body water (TBW), intra (ICF) and extracellular (ECF) fluids, whole-body resistance (R) and reactance (Xc) and PhA. Upper and lower body muscle strength were also assessed. RESULTS: From baseline to after training muscle strength, ALST and PhA significantly (P<0.05) increased. In detraining, significant (P<0.05) reductions in muscle strength, TBW, ECF, ICF and PhA along with significant (P<0.05) increases in R were observed, with the greatest magnitude observed for PhA (Δ=-7.6%). From detraining to retraining a significant reduction in FM along with increases in Xc, PhA and muscle strength were observed. Although an increase was observed from detraining to retraining in PhA, the values were still lower than baseline PhA. CONCLUSIONS: In untrained older women, a RT is associated with increases in PhA, whereas detraining results in a marked decrease in PhA, and more time may be required in retraining to counteract the negative influence of absence of exercise stimulus.

Single and combined effects of body composition phenotypes on carotid intima-media thickness.

BACKGROUND: Central fatness might be a more sensitive predictor of atherosclerotic changes in children than are total body fat measures. However, it is unclear whether a total body fat measure coupled with an estimate of a more central pattern of fat accumulation predicts increased carotid intima-media-thickness (cIMT) better than either measure alone. OBJECTIVE: The objective of the study is to identify the ability of a combination of simple anthropometric screening tools or a combination of objective measures of body composition to predict cIMT. METHODS: cIMT was assessed on the common carotid artery in 349 children aged 11-12 years old (183 girls). Body mass index (BMI), waist circumference (WC) and waist-to-height ratio (WHtR) were dichotomized according to established criteria and indices of total body (TBFI) and abdominal (ABFMI) fat were assessed by dual-energy X-ray absorptiometry and categorized (increased risk ≥85%). Single and combined associations among anthropometric and laboratorial measures with the risk of having increased cIMT (≥85%) and discriminatory performance were tested with logistic regression analysis and Receiver Operator Curve analysis. RESULTS: Children with higher total fatness (BMI and TBFI) or higher central pattern of fat accumulation (WC, WHtR and BFMI) were in higher risk for increased cIMT [odds ratio (OR): 2.08-3.24). The risk for increased cIMT was not higher among children who coupled high total and high central fatness (OR: 2.27-3.10). CONCLUSIONS: Combination of total and central measures of fat does not improve the prediction of increased cIMT in children. Simple surrogate measures of fatness can be used to predict increased cIMT urging special attention to those children who exhibit increased abdominal fat.

Accuracy of a combined heart rate and motion sensor for assessing energy expenditure in free-living adults during a double-blind crossover caffeine trial using doubly labeled water as the reference method.

BACKGROUND/OBJECTIVES: A combined heart rate (HR) and motion sensor (Actiheart) has been proposed as an accurate method for assessing total energy expenditure (TEE) and physical activity energy expenditure (PAEE). However, the extent to which factors such as caffeine may affect the accuracy by which the estimated HR-related PAEE contribution will affect TEE and PAEE estimates is unknown. Therefore, we examined the validity of Actiheart in estimating TEE and PAEE in free-living adults under a caffeine trial compared with doubly labeled water (DLW) as reference criterion. SUBJECTS/METHODS: Using a double-blind crossover trial (Clinicaltrials.gov ID: #NCT01477294) with two conditions (4-day each with a 3-day-washout period), randomly ordered as caffeine (5 mg/kg per day) and placebo (malt-dextrine) intake, TEE was measured by DLW in 17 physically active men (20-38 years) who were non-caffeine users. In each condition, resting energy expenditure (REE) was assessed by indirect calorimetry and PAEE was calculated as (TEE-(REE+0.1 TEE)). Simultaneously, PAEE and TEE were estimated by Actiheart using an individual calibration (ACC+HRstep). RESULTS: Under caffeine, ACC+HRstep explained 76 and 64% of TEE and PAEE from DLW, respectively; corresponding results for the placebo condition were 82 and 66%. No mean bias was found between ACC+HRstep and DLW for TEE (caffeine:-468 kJ per day; placebo:-407 kJ per day), although PAEE was slightly underestimated (caffeine:-856 kJ per day; placebo:-1147 kJ per day). Similar limits of agreement were observed in both conditions ranging from -2066 to 3002 and from -3488 to 1776 kJ per day for TEE and PAEE, respectively. CONCLUSIONS: Regardless of caffeine intake, the combined HR and motion sensor is valid for estimating free-living energy expenditure in a group of healthy men but is less accurate for an individual assessment.

ATP is released from nerve terminals and from activated muscle fibres on stimulation of the rat phrenic nerve.

Nerve stimulation increases the concentration of ATP in the synaptic cleft, which can act as a neurotransmitter or as a presynaptic neuromodulator. Using the luciferin-luciferase assay, we observed that the extracellular concentration of ATP increased by 11-26 nM over a basal concentration of 6 nM, in a frequency dependent manner (1-5 Hz), in the adult rat phrenic nerve-hemidiaphragm preparation. This ATP release depends on nerve activity since it was abolished by tetrodotoxin (1 microM) and is strictly dependent on the presence of extracellular calcium. However, more than half of this nerve-evoked release of ATP is derived from activated muscle fibres since the selective post-synaptic nicotinic receptor antagonist, alpha-bungarotoxin (1 microM), inhibited by over 60% the evoked release of ATP. The presently observed post-synaptic release of ATP together with the previously reported lack of post-synaptic effects of ATP and to the ability of ATP to act as a presynaptic modulator open the possibility that ATP may behave as a retrograde messenger at this neuromuscular junction.

Outbreak of acute gastroenteritis in young children with death due to rotavirus genotype G9 in Rio Branco, Brazilian Amazon region, 2005.

BACKGROUND: An epidemic of acute gastroenteritis occurred in Rio Branco City, Acre State, in Brazil's Amazon region in 2005. An investigation was conducted to confirm the etiology and identify possible risk factors for death. METHODS: Rio Branco municipality surveillance data for the period May to October 2005 were reviewed. In a case-control study, children who died following acute gastroenteritis were compared to age-matched controls with acute gastroenteritis who survived. Rotavirus A (RV-A) was investigated in 799 stool samples and genotyped by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The cumulative incidence of diarrhea in children aged <5 years was 21%. A fatal outcome was significantly associated with uncovered household water storage containers. RV-A was identified in 88% of samples and G9 was the prevalent genotype (71%). CONCLUSIONS: Oral rehydration solution and boiling or chlorinating drinking water likely limited mortality. This epidemic was caused by RV-A genotype G9. After the outbreak, a rotavirus vaccine was introduced into the official childhood immunization schedule in Brazil.