Recent dynamics in Neisseria gonorrhoeae genomic epidemiology in Brazil: antimicrobial resistance and genomic lineages in 2017-20 compared to 2015-16.

OBJECTIVES: Regular quality-assured WGS with antimicrobial resistance (AMR) and epidemiological data of patients is imperative to elucidate the shifting gonorrhoea epidemiology, nationally and internationally. We describe the dynamics of the gonococcal population in 11 cities in Brazil between 2017 and 2020 and elucidate emerging and disappearing gonococcal lineages associated with AMR, compare to Brazilian WGS and AMR data from 2015 to 2016, and explain recent changes in gonococcal AMR and gonorrhoea epidemiology. METHODS: WGS was performed using Illumina NextSeq 550 and genomes of 623 gonococcal isolates were used for downstream analysis. Molecular typing and AMR determinants were obtained and links between genomic lineages and AMR (determined by agar dilution/Etest) examined. RESULTS: Azithromycin resistance (15.6%, 97/623) had substantially increased and was mainly explained by clonal expansions of strains with 23S rRNA C2611T (mostly NG-STAR CC124) and mtr mosaics (mostly NG-STAR CC63, MLST ST9363). Resistance to ceftriaxone and cefixime remained at the same levels as in 2015-16, i.e. at 0% and 0.2% (1/623), respectively. Regarding novel gonorrhoea treatments, no known zoliflodacin-resistance gyrB mutations or gepotidacin-resistance gyrA mutations were found. Genomic lineages and sublineages showed a phylogenomic shift from sublineage A5 to sublineages A1-A4, while isolates within lineage B remained diverse in Brazil. CONCLUSIONS: Azithromycin resistance, mainly caused by 23S rRNA C2611T and mtrD mosaics/semi-mosaics, had substantially increased in Brazil. This mostly low-level azithromycin resistance may threaten the recommended ceftriaxone-azithromycin therapy, but the lack of ceftriaxone resistance is encouraging. Enhanced gonococcal AMR surveillance, including WGS, is imperative in Brazil and other Latin American and Caribbean countries.

Aetiological molecular identification of sexually transmitted infections that cause urethral discharge syndrome and genital ulcer disease in Brazilian men: a nationwide study.

BACKGROUND: Little is known about the aetiology of urethral discharge syndrome (UDS) and genital ulcer disease (GUD) in Brazil due to limited access to laboratory tests and treatment based mainly on the syndromic approach. OBJECTIVES: To update Brazilian treatment guidelines according to the current scenario, the first nationwide aetiological study for UDS and GUD was performed. METHODS: Male participants with urethral discharge (UD) and/or genital ulcer (GU) reports were enrolled. Sample collection was performed by 12 sentinel sites located in the five Brazilian regions. Between 2018 and 2020, 1141 UD and 208 GU samples were collected in a Universal Transport Medium-RT (Copan). A multiplex quantitative PCR kit (Seegene) was used to detect UD: Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), M. hominis (MH), Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV), Ureaplasma parvum (UP), U. urealyticum (UU) and another kit to detect GU: cytomegalovirus (CMV), Haemophilus ducreyi (HD), herpes simplex virus type 1 (HSV1), herpes simplex virus type 2 (HSV2), lymphogranuloma venereum (LGV), Treponema pallidum (TP) and varicella-zoster virus (VZV). RESULTS: In UD samples, the frequency of pathogen detection was NG: 78.38%, CT: 25.6%, MG: 8.3%, UU: 10.4%, UP: 3.5%, MH: 3.5% and TV: 0.9%. Coinfection was assessed in 30.9% of samples, with 14.3% of NG/CT coinfection. The most frequent pathogen identified in GU was HSV2, present in 40.8% of the samples, followed by TP at 24.8%, LGV and CMV at 1%, and HSV1 at 0.4%. Coinfection of TP/HSV2 was detected in 4.4% of samples. VZV and HD were not detected. In 27.7% of the GU samples, no pathogen was detected. CONCLUSION: This study provided the acquisition of unprecedented data on the aetiology of UDS and GUD in Brazil, demonstrated the presence of a variety of pathogens in both sample types and reaffirmed the aetiologies known to be most prevalent globally.

Probing the Electronic Structure of Prussian Blue and Analog Films by Photoemission and Electron Energy Loss Spectroscopies.

X-ray photoelectron spectroscopy (XPS) and reflection electron energy loss spectroscopy (REELS) were employed to characterize the electronic properties of Prussian blue (PB) and its analogs when electrodeposited over metal-decorated carbon nanotubes (CNTs). Through an investigation of the influence of carbon nanotubes (CNTs) and preparation conditions on the electronic structure, valuable insights were obtained regarding their effects on electrochemical properties. XPS analysis enabled the probing of the chemical composition and oxidation states of the film materials, unveiling synthesis-driven variations in their electronic properties. REELS provided information on energy loss and electronic transitions, enabling further characterization of the changes in the electronic structure induced by different preparation methods. Such findings emphasize the importance of surface characterization to understand how the unique electronic properties of such materials can be harnessed to enhance their performance and functionality.

Dilated cardiomyopathy phenotype in Callithrix penicillata (E. Geoffroy, 1812): Case report.

Dilated cardiomyopathy (DCM) is a disease of the heart muscle diagnosed by alterations resulting from ventricular systolic dysfunction with enlargement of the heart chambers, which is still underdiagnosed in non-human primates. This report is the first case of the DCM phenotype diagnosed by echocardiography and confirmed by necropsy in Callithrix penicillata.

First report of spontaneous hepatic fibrosarcoma in a white-tufted marmoset (Callithrix jacchus) kept in captivity.

Reports of sarcomas in nonhuman primates are scarce and, specifically, primary hepatic sarcomas are rare, as in humans. This is the first report of an aggressive hepatic fibrosarcoma in a young adult Callithrix jacchus. The final postmortem diagnosis was obtained by means of immunohistochemical analysis, which confirmed the tumor histogenesis.

Contribution of ultrasound examination in diagnosing platynosomiasis and correlation with macro and microscopic findings in callitrichids kept under human care.

BACKGROUND: Platynosomiasis in non-human primates kept under human care causes chronic disease of the bile ducts and liver, which initially presents with nonspecific signs and can culminate in the death of the animal. Diagnosing this disease is a challenge, and an ultrasound examination can be an excellent tool when it is suspected. METHODS: This study describes the ultrasound findings from 57 marmosets with suspected infection by Platynosomum sp., the correlated hepatobiliary changes, and the anatomopathological findings that confirmed the occurrence of platynosomiasis. RESULTS: In six marmosets (one C. aurita, two C. jacchus, and three Callithrix sp.), Platynosomum infection was confirmed macroscopically (presence of adult trematodes in the gallbladder) and microscopically (adults, larvae, and eggs in histological examinations and eggs in bile and feces). These findings were compatible with the hepatobiliary changes and with images suggestive of parasitic structures in ante-mortem assessments. CONCLUSION: Ultrasound examination demonstrated its usefulness within the clinical routine for investigating this parasitosis.

X-ray induced ultrafast charge transfer in thiophene-based conjugated polymers controlled by core-hole clock spectroscopy.

We explore ultrafast charge transfer (CT) resonantly induced by hard X-ray radiation in organic thiophene-based polymers at the sulfur K-edge. A combination of core-hole clock spectroscopy with real-time propagation time-dependent density functional theory simulations gives an insight into the electron dynamics underlying the CT process. Our method provides control over CT by a selective excitation of a specific resonance in the sulfur atom with monochromatic X-ray radiation. Our combined experimental and theoretical investigation establishes that the dominant mechanism of CT in polymer powders and films consists of electron delocalisation along the polymer chain occurring on the low-femtosecond time scale.

Action of Carduus marianus 6cH on Hepatopathy and Gallstone in Ambystoma mexicanum (Axolotl): Case Report.

BACKGROUND: Carduus marianus is a Mediterranean plant used for treating liver diseases. One of its properties is that it inhibits fat peroxidation, which can contribute toward the prevention of metabolites that are harmful to the liver. OBJECTIVE: The aim of this study was to describe the treatment of fatty liver disease in an axolotl (Ambystoma mexicanum). METHODS: An approximately 6-year-old female specimen of A. mexicanum, of length 21 cm and weight 153 g, presented edema in the rostral region and inappetence. Ultrasound showed hepatopathy and a gallbladder stone. A globule of C. marianus 6cH was diluted in 30 mL distilled water and administered in spray form over the animal twice a day. RESULTS: Twenty days after the beginning of the treatment, improvement of the facial edema and return of appetite were observed. Ultrasound showed improvement in the appearance of the liver and reduction in the size of the gallstone. The Modified Naranjo Criteria for Homeopathy score was +9 in this case, thus suggesting a causal relationship between the use of homeopathic medicine and the clinical outcome. CONCLUSION: C. marianus 6cH appeared to be effective in treating fatty liver disease and gallstone in this specimen of A. mexicanum.

Nanoencapsulation of R-phycoerytrin extracted from Solieria filiformis improves protein stability and enables its biological application as a fluorescent dye.

We aimed to encapsulate R-PE to improve its stability for use as a fluorescent probe for cancer cells. Purified R-PE from the algae Solieria filiformis was encapsulated in polymeric nanoparticles using PCL. Nanoparticles were characterised and R-PE release was evaluated. Also, cellular uptake using breast and prostate cancer cells were performed. Nanoparticles presented nanometric particle size (198.8 ± 0.06 nm) with low polydispersity (0.13 ± 0.022), negative zeta potential (-18.7 ± 1.10 mV), and 50.0 ± 7.3% encapsulation. FTIR revealed that R-PE is molecularly dispersed in PCL. DSC peak at 307 °C indicates the presence of R-PE in the nanoparticle. Also, in vitro, it was demonstrated low release for nanoparticles and degradation for the free R-PE. Finally, cellular uptake demonstrated the potential of R-PE/PCL nanoparticles for cancer cell detection. Nanoparticles loaded with R-PE can overcome instability and allow application as a fluorescent probe for cancer cells.

Action of Folliculinum 6cH on Folliculogenesis in the Snake Python Regius (Shaw, 1802) - Two Case Reports.

BACKGROUND: Folliculinum is a homeopathic sarcode medication that is mainly used for regulating the estrous cycle and reproductive function. In snakes, it is common to observe low frequencies of reproduction. Ovulation is related to follicular size and morphology, and follicle homogeneity may indicate the absence of problems such as dystocia (egg retention) or follicular stasis. OBJECTIVE: The objective of the present study was to report on follicular stage changes in two ball pythons, Python regius (Shaw, 1802), which were treated using Folliculinum 6cH in a zoological park setting. METHODS: Two female pythons - one weighing 2.8 kg, the other weighing 2.5 kg, and neither with a history of reproduction - were examined by ultrasound to enable viewing of ovarian follicles in different phases and sizes. Folliculinum 6cH, two globules diluted in 200 mL of water, was administered, targeted to the eyes and nostril regions. RESULTS: Even given the slow metabolism of reptiles, ultrasound revealed an improvement in follicle homogeneity between 6 and 15 days after the start of homeopathy in both snakes; there was also improved weight gain in both animals. The MOdified NARanjo Criteria for Homeopathy (MONARCH) score was +8 in each of the cases, suggesting a causal relationship between the use of homeopathic medicine and clinical outcome. CONCLUSION: Folliculinum 6cH may have promoted stabilization of follicle size and enabled folliculogenesis in two ball pythons.

First report on herpesvirus in black-fronted titi (Callicebus nigrifrons) kept under human care.

Keeping Neotropical primates in captivity puts them at great risk of illness because of their susceptibility to human herpesvirus. This is the first report on herpesvirus in Callicebus nigrifrons that developed clinical disease and was confirmed by immunohistochemical and RT-PCR. Diagnosis and prevention are essential for the conservation of species.

Electron delocalisation in conjugated sulfur heterocycles probed by resonant Auger spectroscopy.

We propose a novel approach for an indirect probing of conjugation and hyperconjugation in core-excited molecules using resonant Auger spectroscopy. Our work demonstrates that the changes in the electronic structure of thiophene (C4H4S) and thiazole (C3H3NS), occurring in the process of resonant sulfur K-shell excitation and Auger decay, affect the stabilisation energy resulting from π-conjugation and hyperconjugation. The variations in the stabilisation energy manifest themselves in the resonant S KL2,3L2,3 Auger spectra of thiophene and thiazole. The comparison of the results obtained for the conjugated molecules and for thiolane (C4H8S), the saturated analogue of thiophene, has been performed. The experimental observations are interpreted using high-level quantum-mechanical calculations and the natural bond orbital analysis.

Long-term ethanol intake causes morphological changes in Schistosoma mansoni adult worms in mice.

Schistosoma mansoni adult worms are extensively challenged by reactive oxygen species from intrinsic sources. However, the effects of extrinsic sources such as ethanol have not been looked at in schistosomes. We examined adult worms recovered from ethanol-consuming mice by light (LM), confocal (CM) and scanning electron microscopy (SEM) to address this question. Schistosomiasis-infected mice were orally gavaged with 18% (v/v) ethanol from 35 to 63 days post-infection, when they were euthanized. CM examination revealed reduced germ cells density (-36%, p = 0.0001) and sperm density (-58%, p = 0.0001) in testicular lobes, and immature cells in seminal vesicle compared to unexposed control worms. Female worms showed reduced density of vitellin glands (-71%, p = 0.0001), maturation of oocytes (-7%, p = 0.0071) and reduced spermatozoa density (-23%, p = 0.0002) within the seminal receptacle. SEM revealed remarkable damages in male's tegument, including tubercles flattening, tegumental peeling and erosive lesions. Given that lipids are present in reproductive system and tegument, our results suggest that phenotypic changes are due to ethanol-induced lipid peroxidation. To the best of our knowledge, this is the first report revealing the biological action of ethanol intake on adult schistosomes in vivo.

Physiological responses to interval endurance exercise at different levels of blood flow restriction.

PURPOSE: We aimed to identify a blood flow restriction (BFR) endurance exercise protocol that would both maximize cardiopulmonary and metabolic strain, and minimize the perception of effort. METHODS: Twelve healthy males (23 ± 2 years, 75 ± 7 kg) performed five different exercise protocols in randomized order: HI, high-intensity exercise starting at 105% of the incremental peak power (P peak); I-BFR30, intermittent BFR at 30% P peak; C-BFR30, continuous BFR at 30% P peak; CON30, control exercise without BFR at 30% P peak; I-BFR0, intermittent BFR during unloaded exercise. Cardiopulmonary, gastrocnemius oxygenation (StO2), capillary lactate ([La]), and perceived exertion (RPE) were measured. RESULTS: V̇O2, ventilation (V̇ E), heart rate (HR), [La] and RPE were greater in HI than all other protocols. However, muscle StO2 was not different between HI (set1-57.8 ± 5.8; set2-58.1 ± 7.2%) and I-BRF30 (set1-59.4 ± 4.1; set2-60.5 ± 6.6%, p < 0.05). While physiologic responses were mostly similar between I-BFR30 and C-BFR30, [La] was greater in I-BFR30 (4.2 ± 1.1 vs. 2.6 ± 1.1 mmol L-1, p = 0.014) and RPE was less (5.6 ± 2.1 and 7.4 ± 2.6; p = 0.014). I-BFR30 showed similar reduced muscle StO2 compared with HI, and increased blood lactate compared to C-BFR30 exercise. CONCLUSION: Therefore, this study demonstrate that endurance cycling with intermittent BFR promotes muscle deoxygenation and metabolic strain, which may translate into increased endurance training adaptations while minimizing power output and RPE.

Mechanism of Action of TiF4 on Dental Enamel Surface: SEM/EDX, KOH-Soluble F, and X-Ray Diffraction Analysis.

This in vitro study aimed to evaluate the action of TiF4 on sound and carious bovine and human enamel. Sound (S) and pre-demineralised (DE) bovine and human (primary and permanent) enamel samples were treated with TiF4 (pH 1.0) or NaF varnishes (pH 5.0), containing 0.95, 1.95, or 2.45% F for 12 h. The enamel surfaces were analysed using SEM-EDX (scanning electron microscopy/energy-dispersive X-ray spectroscopy) (n = 10, 5 S and 5 DE) and KOH-soluble fluoride was quantified (n = 20, 10 S and 10 DE). Hydroxyapatite powder produced by precipitation method was treated with the corresponding fluoride solutions for 1 min (n = 2). The formed compounds were detected using X-ray diffraction (XRD). All TiF4 varnishes produced a coating layer rich in Ti and F on all types of enamel surface, with micro-cracks in its extension. TiF4 (1.95 and 2.45% F) provided higher fluoride deposition than NaF, especially for bovine enamel (p < 0.0001). It also induced a higher fluoride deposition on DE samples compared to S samples (p < 0.0001), except for primary enamel. The Ti content was higher for bovine and human primary enamel than human permanent enamel, with some differences between S and DE. The XRD analysis showed that TiF4 induced the formation of new compounds such as CaF2, TiO2, and Ti(HPO4)2·H2O. In conclusion, TiF4 (>0.95% F) interacts better, when compared to NaF, with bovine and human primary enamel than with human permanent enamel. TiF4 provoked higher F deposition compared to NaF. Carious enamel showed higher F uptake than sound enamel by TiF4 application, while Ti uptake was dependent on the enamel condition and origin.

Evaluation of antioxidant activity of grapevine leaves extracts (Vitis labrusca) in liver of Wistar rats.

The aim of this study was to evaluate the hepatoprotection of organic and conventional grapevine leaves extract (Vitis labrusca). The total polyphenol content and the isolate polyphenols by HPLC were evaluate. The animals received intraperitoneal injections of saline or extracts (conventional or organic - 30 mg/kg) for 14 days. On day 15, the rats received carbon tetrachloride (CCl4) or mineral oil (i.p.). After 4h, the animals were euthanized. The analysis of the liver enzymes activity (AST, ALT, GGT) was performed using serum, obtained by blood and the levels of lipid peroxidation (TBARS), protein oxidation (carbonyl), and the activity of antioxidant enzymes superoxide dismutase and catalase were analyzed in the liver. The results showed that the organic extract is richer in polyphenol and resveratrol than the conventional one. Both extracts prevent lipid peroxidation and protein oxidation generated by CCl4. Moreover, the extracts demonstrated ability to modulate the activity of SOD and CAT, as well as to establish a balance in the ratio of SOD/CAT. We also found that the CCl4 increased the levels of AST and GGT, and that both extracts prevent this. These results indicate that grapevine leaves extracts, both, organic and conventional, can prevent liver disorders.

Overview of childhood vaccination coverage in Brazil and the impact of the COVID-19 pandemic: Is our children's health at risk? A review of pre-COVID-19 periods and during the COVID-19 pandemic.

Introduction: The coronavirus disease (COVID)-19 has had a great impact on several aspects related to the population's health, including the vaccination adherence rate. This study describes how childhood vaccination coverage (CVC) in Brazil was affected by the pandemic in the period from 2020 to 2022 and explores the relationship between this data and the Human Development Index (HDI), and the number of votes received in the government with a right-wing political ideology. Methods: An ecological analysis of CVC was carried out including 12 vaccines. The HDI was evaluated considering the HDI-General, HDI-Income, HDI-Longevity, and HDI-Education. The percentage of valid votes received by the former president (right-wing political ideology) was also obtained. Spearman correlation tests were applied to compare markers. Results: During the period analyzed, it was observed a linear growth trend in CVC between 2015 and 2018 regarding all vaccines. However, from 2018 onwards, after the presidential elections in Brazil, the CVC reduced significantly, showing an even more pronounced decrease with the start of the COVID-19 pandemic. This reduction in CVC observed for some vaccines was related to the higher percentage of votes for the government with a right-wing political ideology, especially in relation to the BCG (bacillus Calmette and Guerin) and pentavalent (protecting against diphtheria, tetanus, pertussis, hepatitis B, and Haemophilus influenzae type b bacteria) vaccines. In addition, when analyzing the HDI, it was observed that the lowest values of this indicator were associated with a more expressive reduction in CVC, mainly related to yellow fever, pentavalent, 10-valent pneumococcal conjugate, Human rotavirus, and triple viral (protecting against measles, mumps, and rubella - MMR) vaccines. Conclusion: Although Brazil has a successful and exemplary record in combating several diseases, mainly due to the high rate of CVC, the continuous reduction in this coverage must be thoroughly evaluated by health managers.

Beckwith-Wiedemann syndrome mimicking the classical form of congenital adrenal hyperplasia in newborn screening.

Beckwith-Wiedemann syndrome (BWS) is a common genetic congenital disease characterized by somatic overgrowth and its broad clinical spectrum includes pre- and post-natal macrosomia, macroglossia, visceromegaly, increased risk of neonatal hypoglycemia, and development of embryonic tumors. BWS occurs due to genetic/epigenetic changes involving growth-regulating genes, located on region 11p15, with an important genotype-phenotype correlation. Congenital adrenal hyperplasia (CAH) comprises a spectrum of autosomal recessive diseases presenting a variety of clinical manifestations due to a deficiency in one of the enzymes involved in cortisol secretion. Early diagnosis based on newborn screening prevents the adrenal crisis and early infant death. However, high 17-hydroxyprogesterone (17-OHP) levels can occur in newborns or premature infants without CAH, in situations of stress due to maternal or neonatal factors. Here, we report new cases of false-positive diagnosis of 21-hydroxylase deficiency during newborn screening - two girls and one boy with BWS. Methylation-specific multiplex ligation-dependent probe amplification revealed a gain of methylation in the H19 differentially methylated region. Notably, all three cases showed a complete normalization of biochemical changes, highlighting the transient nature of these hormonal findings that imitate the classical form of CAH. This report sheds light on a new cause of false-positive 21-hydroxylase deficiency diagnosis during newborn screening: Beckwith-Wiedemann syndrome.

COVID-19 in Brazil: a 3-year update.

Three years into the coronavirus disease (COVID)-19 pandemic and the world is still struggling with the aftermath of this global health crisis. In Brazil, we are witnessing serious economic, health, social, and political problems. The rapid spread of the virus in our country was the result of a shortage of vaccines and the lack of an effective national campaign to identify and report cases. This health crisis also intensified social inequalities, hitting Indigenous peoples hard due to the lack of access to health services. In addition, rising unemployment and overcrowding of the health system made contagion possible, especially among the most vulnerable, increasing the number of serious cases of the disease. It is important to highlight that emotional problems worsened, the educational system was severely affected, and domestic violence increased during the confinement period, in addition to the fact that the pandemic exposed the great disparities of regional inequalities that exist across the country, mainly concerning health management.

Cystic Fibrosis: A Descriptive Analysis of Deaths in a Two-Decade Period in Brazil According to Age, Race, and Sex.

The diagnosis of cystic fibrosis has improved in the last few years due to greater access to diagnostic tools and the evolution of molecular biology; the knowledge obtained has contributed to the understanding of its death profile. In this context, an epidemiological study was developed focusing on deaths from cystic fibrosis in Brazil from 1996 to 2019. The data were collected from the Data-SUS (Unified National Health System Information Technology Department from Brazil). The epidemiological analysis included patients' age groups, racial groups, and sex. In our data, between 1996 and 2019, Σ3050 deaths were recorded, totaling a ≅330% increase in the number of deaths resulting from cystic fibrosis. This fact might be related to a better diagnosis of the disease, mainly in patients from racial groups that are not commonly associated with cystic fibrosis, such as Black individuals, Hispanic or Latino (mixed individuals/Pardos) individuals, and American Indians (Indigenous peoples from Brazil). Regarding of race, the Σ of deaths was: nine (0.3%) in the American Indian group, 12 (0.4%) in the Asian group, 99 (3.6%) in the Black or African American group, 787 (28.6%) in the Hispanic or Latino group, and 1843 (67.0%) in the White group. The White group showed the highest prevalence of deaths, and the increase in mortality was ≅150 times in this group, while, in the Hispanic or Latino group, it was ≅75 times. Regarding sex, the numbers and percentage of deaths of both male (N = 1492; 48.9%) and female (N = 1557; 51.1%) patients were seen to be relatively close. As for age groups, the >60-year-old group presented the most significant results, with an increase of ≅60 times in the registered deaths. In conclusion, in Brazil, despite the number of deaths from cystic fibrosis being prevalent in the White group, it increased in all racial groups (Hispanic or Latino, Black or African American, American Indian, or Asian individuals) and was associated with older age.