Target Identification in Anti-Tuberculosis Drug Discovery.

Mycobacterium tuberculosis (Mtb) is the etiological agent of tuberculosis (TB), a disease that, although preventable and curable, remains a global epidemic due to the emergence of resistance and a latent form responsible for a long period of treatment. Drug discovery in TB is a challenging task due to the heterogeneity of the disease, the emergence of resistance, and uncomplete knowledge of the pathophysiology of the disease. The limited permeability of the cell wall and the presence of multiple efflux pumps remain a major barrier to achieve effective intracellular drug accumulation. While the complete genome sequence of Mtb has been determined and several potential protein targets have been validated, the lack of adequate models for in vitro and in vivo studies is a limiting factor in TB drug discovery programs. In current therapeutic regimens, less than 0.5% of bacterial proteins are targeted during the biosynthesis of the cell wall and the energetic metabolism of two of the most important processes exploited for TB chemotherapeutics. This review provides an overview on the current challenges in TB drug discovery and emerging Mtb druggable proteins, and explains how chemical probes for protein profiling enabled the identification of new targets and biomarkers, paving the way to disruptive therapeutic regimens and diagnostic tools.

Influence of Gender and Age of Brown Seaweed (Fucus vesiculosus) on Biochemical Activities of Its Aqueous Extracts.

Fucus vesiculosus L. is a common coastal brown seaweed associated with various benefits to human health due to its phenolic content and nutrients and is used as food through different methods of consumption. This study aims to evaluate the influence of the seaweed's gender and growth stage on different types of biological activities as well as its chemical constitution and elements present. Akin to food preparation, aqueous extracts of the seaweed were prepared at 25 °C (salad) and 100 °C (soup). Biological activities were determined by measuring total phenol content (TPC), antioxidant activity and inhibition of acetylcholinesterase (AChE). Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS/MS) was used for compound identification, and elemental analysis was carried out by using Total Reflection X-ray Fluorescence Spectrometry (TXRF). Older females and males had higher TPC compared to the new ones at 100 °C. Antioxidant activity depended on the extraction temperature but was higher for the youngest male at 100 °C. AChE inhibitory activity was higher for older males at 25 °C, but at 100 °C it was higher for older females. Primary metabolites and various phloroglucinol were the main compounds identified. Additionally, since this seaweed is often harvested in estuarine systems with high anthropogenic impacts, its safety was evaluated through the evaluation of the sample's metal content. The heavy metals detected are within the limits established by various regulating entities, pointing to a safe food source.

The Impact of the Philtrum on Facial Attractiveness, Perioral Proportions, and Perceptions of Facial Aging.

BACKGROUND: Lip enhancement continues to be one of the most popular facial procedures. State-of-the-art lip enhancement involves sculpting of perilabial soft tissue, in particular, the philtrum, but it is unclear how patients perceive their philtral contours. OBJECTIVES: The aim of this study was to test the impact of the philtrum on attractiveness, and on the perception of facial proportions and age. METHODS: Respondents (1100, including 50 aesthetic specialists) were presented with 2 sets of identical portraits, 1 set of a young Caucasian female and 1 set of an aged Caucasian female, with 1 portrait in each set having an upper lip lacking any philtral contours. The respondents were asked to choose which upper lip was more attractive, appeared longer, and looked older. Rankings were analyzed according to population demographics. RESULTS: Overall, most respondents considered the young face (84%) and the aged face (68%) with philtral contours to be more attractive. Moreover, the majority of respondents (81%) considered the upper lip of the young face without a philtrum to appear longer in a craniocaudal orientation (67% for the aged image), and 67% of respondents described the image of the young woman with no philtral definition as appearing older (55% for the aged face with no philtrum). CONCLUSIONS: This study confirms the authors' hypothesis that the existence of philtral contours exerts a significant impact on perioral attractiveness, and that its absence can be related to a longer appearance of the upper lip as well as an older facial appearance.

Fish biodiversity of Saint Peter and Saint Paul's Archipelago, Mid-Atlantic Ridge, Brazil: new records and a species database.

Saint Peter and Saint Paul's Archipelago (SPSPA), one of the smallest and most isolated island groups in the world, is situated on the Mid-Atlantic Ridge, between Brazil and the African continent. SPSPA has low species richness and high endemism; nonetheless, the diversity of fishes from deep habitats (>30 m depth) had not been previously studied in detail. Several expeditions conducted between 2009 and 2018 explored the shallow and deep reefs of SPSPA using scuba, closed-circuit rebreathers, manned submersibles, baited remote underwater stereo-videos (stereo-BRUV) and fishing between 0 and 1050 m depth. These expeditions yielded 41 new records of fishes for SPSPA: 9 in open waters, 9 in shallow waters (0-30 m), 8 in mesophotic ecosystems (30-150 m) and 15 in deeper reefs (>150 m). Combined with literature records of adult pelagic, shallow and deep-reef species, as well as larvae, the database of the fish biodiversity for SPSPA currently comprises 225 species (169 recorded as adult fishes and 79 as larvae, with 23 species found in both stages). Most of them (112) are pelagic, 86 are reef-associated species and 27 are deep-water specialists. Species accumulation curves show that the number of fish species has not yet reached an asymptote. Whereas the number of species recorded in SPSPA is similar to that in other oceanic islands in the Atlantic Ocean, the proportion of shorefishes is relatively lower, and the endemism level is the third highest in the Atlantic. Twenty-nine species are listed as threatened with extinction. Observations confirm the paucity of top predators on shallow rocky reefs of the island, despite the presence of several pelagic shark species around SPSPA. Because all of the endemic species are reef associated, it is argued that the new marine-protected areas created by the Brazilian government do not ensure the protection and recovery of SPSPA's biodiversity because they allow exploitation of the most vulnerable species around the archipelago itself. This study suggests a ban on reef fish exploitation inside an area delimited by the 1000 m isobath around the islands (where all known endemics are concentrated) as the main conservation strategy to be included in the SPSPA management plan being prepared by the Brazilian government.

Review of the Brazilian species of Physiculus (Gadiformes: Moridae), with description of a new species from Saint Peter and Saint Paul Archipelago, equatorial Atlantic.

Three valid species of the genus Physiculus are known from the Brazilian marinewaters. A fourth, new species, Physiculus cirm n. sp.., is described based on seventeen specimens collected in the surroundings of Saint Peter and Saint Paul Archipelago, equatorial Atlantic. A review of the Brazilian species of Physiculus is provided, as well as a key to the species of the genus reported from the Atlantic Ocean. The new species is distinguished from all its congeners, except P. cynodon and P. karrerae, by the large number of longitudinal series of scales (156-189 vs. 70-150). P. cynodon from the Northern Pacific has about 200 longitudinal series of scales, and it differs from the new species by the number of rays of the first dorsal fin (6-8 vs. 10 in P. cynodon), pectoral-fin rays (20-25 vs. 27 in P. cynodon), and the presence of an outer row of large canine teeth on upper and lower jaw. From P. karrerae, which has 134-160 longitudinal series of scales, the new species differs by the presence of scales on the tip of the snout and dorsal-fin membrane, and the number of pectoral-fin rays (20-25 vs. 24-27 in P. karrerae). [Zoobank URL: urn:lsid:zoobank.org:act:1AFBC251-2BB1-4479-98A4-307188EC5D66].

"The Worst Part Was Coming Back Home and Feeling Like Crying": Experiences of Lesbian, Gay, Bisexual and Trans Students in Portuguese Schools.

Portugal is one of the most egalitarian countries in Europe in terms of lesbian, gay, bisexual, transgender, and intersex (LGBTI) individuals' legal rights. However, regarding education Portugal still lacks specific policies, plans and interventions to protect LGBTI students. To assess the perceptions of self-identified LGBTI youth regarding their school context, a total of 663 participants (aged from 15 to 20 years old) filled in an on-line questionnaire about their school climate. One hundred and forty-six of them answered an open-ended question about their personal experiences. A thematic analysis of these answers was conducted, and four main categories were identified: (i) victimization, (ii) coming out experiences, (iii) support networks, and (iv) demands. Most participants reported experiences of discrimination, and several sources of prejudice were identified. Furthermore, participants also recognized a lack of LGBTI information in school curriculum and made several demands. Besides inclusive laws, we suggest that the safety and the well-being of LGBTI youths in Portuguese schools depend upon others measures, such as teacher and school staff training, curricula inclusive of LGBTI diversity, and local strategies, such as Gay-Straight Alliances.

Exploring D-xylose oxidation in Saccharomyces cerevisiae through the Weimberg pathway.

Engineering of the yeast Saccharomyces cerevisiae towards efficient D-xylose assimilation has been a major focus over the last decades since D-xylose is the second most abundant sugar in nature, and its conversion into products could significantly improve process economy in biomass-based processes. Up to now, two different metabolic routes have been introduced via genetic engineering, consisting of either the isomerization or the oxido-reduction of D-xylose to D-xylulose that is further connected to the pentose phosphate pathway and glycolysis. In the present study, cytosolic D-xylose oxidation was investigated instead, through the introduction of the Weimberg pathway from Caulobacter crescentus in S. cerevisiae. This pathway consists of five reaction steps that connect D-xylose to the TCA cycle intermediate α-ketoglutarate. The corresponding genes could be expressed in S. cerevisiae, but no growth was observed on D-xylose indicating that not all the enzymes were functionally active. The accumulation of the Weimberg intermediate D-xylonate suggested that the dehydration step(s) might be limiting, blocking further conversion into α-ketoglutarate. Although four alternative dehydratases both of bacterial and archaeon origins were evaluated, D-xylonate accumulation still occurred. A better understanding of the mechanisms associated with the activity of dehydratases, both at a bacterial and yeast level, appears essential to obtain a fully functional Weimberg pathway in S. cerevisiae.

Intermediate osteotomies in rhinoplasty: a new perspective.

The nasal dorsum framework plays a major role in nasal and facial harmony. This study presents a new approach to an already known technique, intermediate osteotomies, which can be used routinely in patients with cosmetically unpleasant changes in eyebrow-tip line features, allowing the acquisition of a natural, aesthetically agreeable and harmonious dorsum contour, while preserving nasal function. The approach was evaluated retrospectively in 50 consecutive patients at a tertiary referral hospital. Patients were asked to complete a survey to rate their satisfaction with the cosmetic outcome on a 5-point visual analog scale (VAS) and to compare the impact of nasal obstruction on their quality of life, pre- and postoperatively, by completing the validated Nasal Obstruction Symptom Evaluation (NOSE) scale. Satisfaction was scored as excellent (satisfaction level VAS average score, 4.4), with more than 50% of the patients completely satisfied with the aesthetic result, while NOSE scale scores demonstrated preservation of nasal function postoperatively. Typical complications of the upper third of the nose following osteotomies were not observed.

Effect of nitrogen availability on the poly-3-D-hydroxybutyrate accumulation by engineered Saccharomyces cerevisiae.

Poly-3-D-hydroxybutyrate (or PHB) is a polyester which can be used in the production of biodegradable plastics from renewable resources. It is naturally produced by several bacteria as a response to nutrient starvation in the excess of a carbon source. The yeast Saccharomyces cerevisiae could be an alternative production host as it offers good inhibitor tolerance towards weak acids and phenolic compounds and does not depolymerize the produced PHB. As nitrogen limitation is known to boost the accumulation of PHB in bacteria, the present study aimed at investigating the effect of nitrogen availability on PHB accumulation in two recombinant S. cerevisiae strains harboring different xylose consuming and PHB producing pathways: TMB4443 expressing an NADPH-dependent acetoacetyl-CoA reductase and a wild-type S. stipitis XR with preferential use of NADPH and TMB4425 which expresses an NADH-dependent acetoacetyl-CoA reductase and a mutated XR with a balanced affinity for NADPH/NADH. TMB4443 accumulated most PHB under aerobic conditions and with glucose as sole carbon source, whereas the highest PHB concentrations were obtained with TMB4425 under anaerobic conditions and xylose as carbon source. In both cases, the highest PHB contents were obtained with high availability of nitrogen. The major impact of nitrogen availability was observed in TMB4425, where a 2.7-fold increase in PHB content was obtained. In contrast to what was observed in natural PHB-producing bacteria, nitrogen deficiency did not improve PHB accumulation in S. cerevisiae. Instead the excess available carbon from xylose was shunted into glycogen, indicating a significant gluconeogenic activity on xylose.

Prognostic stratification in pulmonary hypertension: A multi-biomarker approach. / Estratificação prognóstica na hipertensão pulmonar: valor acrescido da abordagem multibiomarcadores.

INTRODUCTION: Pulmonary hypertension (PH) covers a group of conditions characterized by an increase in pulmonary vascular resistance leading to right ventricular failure. Risk stratification is crucial for adequate prognostic and therapeutic assessment. However, the accuracy of conventional parameters is limited, especially biomarkers. OBJECTIVES: To determine the prognostic value of new biomarkers and their combination in a multi-biomarker approach to predict outcome in patients with PH. METHODS: In this prospective cohort study, PH patients underwent clinical, echocardiographic and laboratory assessment, including quantification of serum N-terminal pro-brain natriuretic peptide (NT-proBNP) and of the following new biomarkers: mid-regional pro-adrenomedullin (MR-proADM), copeptin, endothelin-1, mid-regional pro-atrial natriuretic peptide (MR-proANP) and soluble ST2 (sST2), the interleukin-33 receptor. The accuracy of the different parameters for predicting all-cause mortality and death or hospitalization of cardiac causes was determined. The prognostic value of a multi-biomarker score based on the tertile distribution of serum NT-proBNP, MR-proANP, renin and sST2 was compared to conventional markers. RESULTS: Forty-three patients (72.1% female, age 59±15 years) were included, most of whom (65.1%) had group 1 PH. During a median follow-up of 34 months, 26% of the patients died and 35% were hospitalized for cardiac causes. Atrial and ventricular dimensions and right ventricular fractional area change were prognostic predictors. Log NT-proBNP (HR: 31.14; 95% CI: 3.12-310.7; p=0.003) and renin (HR: 1.02; 95% CI: 1.005-1.038; p=0.009) were independent predictors of mortality. MR-proANP (HR: 1.008; 95% CI 1.004-1.011; p<0.001) and sST2 (HR: 1.005; 95% CI 1.001-1.009; p=0.04) were predictors of death or hospitalization. The prognostic value of the multi-biomarker score was higher than any of the conventional parameters, and enabled identification of risk groups (the high-risk group had three-year mortality of 77.8%). CONCLUSION: A multi-biomarker approach was superior for risk stratification to any single marker. A score that incorporates NT-proBNP, MR-proANP, renin and sST2 accurately identifies patients at low, intermediate and high risk.

Anaerobic poly-3-D-hydroxybutyrate production from xylose in recombinant Saccharomyces cerevisiae using a NADH-dependent acetoacetyl-CoA reductase.

BACKGROUND: Poly-3-D-hydroxybutyrate (PHB) that is a promising precursor for bioplastic with similar physical properties as polypropylene, is naturally produced by several bacterial species. The bacterial pathway is comprised of the three enzymes ß-ketothiolase, acetoacetyl-CoA reductase (AAR) and PHB synthase, which all together convert acetyl-CoA into PHB. Heterologous expression of the pathway genes from Cupriavidus necator has enabled PHB production in the yeast Saccharomyces cerevisiae from glucose as well as from xylose, after introduction of the fungal xylose utilization pathway from Scheffersomyces stipitis including xylose reductase (XR) and xylitol dehydrogenase (XDH). However PHB titers are still low. RESULTS: In this study the acetoacetyl-CoA reductase gene from C. necator (CnAAR), a NADPH-dependent enzyme, was replaced by the NADH-dependent AAR gene from Allochromatium vinosum (AvAAR) in recombinant xylose-utilizing S. cerevisiae and PHB production was compared. A. vinosum AAR was found to be active in S. cerevisiae and able to use both NADH and NADPH as cofactors. This resulted in improved PHB titers in S. cerevisiae when xylose was used as sole carbon source (5-fold in aerobic conditions and 8.4-fold under oxygen limited conditions) and PHB yields (4-fold in aerobic conditions and up to 5.6-fold under oxygen limited conditions). Moreover, the best strain was able to accumulate up to 14% of PHB per cell dry weight under fully anaerobic conditions. CONCLUSIONS: This study reports a novel approach for boosting PHB accumulation in S. cerevisiae by replacement of the commonly used AAR from C. necator with the NADH-dependent alternative from A. vinosum. Additionally, to the best of our knowledge, it is the first demonstration of anaerobic PHB synthesis from xylose.

A new species of Anthiadinae (Teleostei: Serranidae) from São Pedro and São Paulo Archipelago, Brazil, Equatorial Atlantic.

A new species of Anthiadinae, Odontanthias cauoh, is described from São Pedro and São Paulo Archipelago (SPSPA), Brazil, Equatorial Atlantic. The new species differs from its only Atlantic congener by a combination of several characters: pectoral-fin rays 20; total gill rakers on first arch 39; vomerine tooth patch with a posterior prolongation; pelvic fin not reaching base of last anal-fin ray; dorsal, anal and pelvic fins scaleless; and coloration (dorsal, anal, and pelvic fins mostly dark red). The record of the new species contributes to increase the fish endemism in SPSPA.

Safety Approach to Otoplasty: A Surgical Algorithm.

An algorithm was developed through an evolution of refinements in surgical technique with the goal to minimize risk and morbidity in otoplasty. Key principles were avoidance of cartilage incisions and transections and the use of multiple surgical techniques to distribute the "surgical load" evenly among these techniques. The present retrospective study was designed to test safety and efficacy of the concept in 100 consecutive patients and to discuss the results in light of the literature. Data detailing the surgery, preoperative, and postoperative period were extracted from the record and during patient interviews. Patients were contacted to complete a questionnaire to rate the postoperative pain and their satisfaction with the final outcome on a 6-point visual analog scale (VAS). An expert and a lay panel assessed preoperative and postoperative frontal-view photographs, using the same VAS. Pain in the postoperative was rated as minor (pain level VAS average score, 2.33) and patients' satisfaction was excellent (satisfaction level VAS average score, 1.82). The assessment by the panels of expert and lay evaluators paralleled these outcomes with a postoperative average VAS score of 1.69 and 1.87, respectively. Cartilage incision and transection can be effectively avoided in otoplasty. Even distribution of the surgical load among multiple techniques avoids the problems associated with "overload" of a single technique. The innovative technique of cortical mastoid drill-out is described. High satisfaction with the results, excellent patient comfort, and a favorable safety profile are associated with the present algorithm.

Engineering of Saccharomyces cerevisiae for the production of poly-3-d-hydroxybutyrate from xylose.

Poly-3-d-hydroxybutyrate (PHB) is a promising biopolymer naturally produced by several bacterial species. In the present study, the robust baker's yeast Saccharomyces cerevisiae was engineered to produce PHB from xylose, the main pentose found in lignocellulosic biomass. The PHB pathway genes from the well-characterized PHB producer Cupriavidus necator were introduced in recombinant S. cerevisiae strains already capable of pentose utilization by introduction of the fungal genes for xylose utilization from the yeast Scheffersomyces stipitis. PHB production from xylose was successfully demonstrated in shake-flasks experiments, with PHB yield of 1.17 ± 0.18 mg PHB g(-1) xylose. Under well-controlled fully aerobic conditions, a titer of 101.7 mg PHB L(-1) was reached within 48 hours, with a PHB yield of 1.99 ± 0.15 mg PHB g(-1) xylose, thereby demonstrating the potential of this host for PHB production from lignocellulose.

Orbital compartment: effects of emergent canthotomy and cantholysis.

The objective of this study was to determine the effect of orbital decompression procedures on the intraocular pressure (IOP). The orbital compartment syndrome represents an emergency situation. Due to the elevated IOP vision loss may ensue. Several maneuvers including lateral canthotomy are discussed to reduce the IOP. Eight orbits were studied in a fresh frozen cadaveric model (4 specimens). Intraorbital volume was determined by CT volumetry. An orbital compartment syndrome was simulated by injecting viscous material into the orbit. Injected volumes were documented and lateral canthotomy, cantholysis, inferior and superior septolysis were performed. IOP and exophthalmometric measurements were obtained after each intervention. Controlled elevation of IOP was achieved in all specimens. IOP was partially reduced after performing a lateral canthotomy in eight orbits. IOP was significantly and sufficiently decreased under 20 mmHg by inferior cantholysis in seven orbits. An additional superior cantholysis was necessary in two orbits to achieve a complete decompression. Inferior or superior septolysis were not needed to further reduce the IOP. Lateral canthotomy must be followed by an inferior cantholysis to successfully decompress an orbital compartment syndrome in the majority of cases. Occasionally, superior cantholysis may generate additional benefit. Additional inferior and superior septolysis were not shown to provide a beneficial effect when performed after canthotomy and cantholysis.

A low-complex multi-channel methodology for noise detection in phonocardiogram signals.

The phonocardiography (PCG) is an important technique for the diagnosis of several heart conditions. However, the PCG signal is highly prone to noise, which can be an obstacle for the detection and interpretation of physiological heart sounds. Thus, the detection and elimination of noise present in PCG signals is crucial for the accurate analysis of heart sounds, especially in p-health environments. Noise can be introduced by various internal factors (e.g., respiration and laughing) and by external conditions (e.g., phone ringing or door closing). To mention also that the noise frequency components are typically overlapped with the PCG spectrum, increasing the complexity of the analysis. The purpose of the present work consists in the detection of noisy periods willfully introduced during the performance of three different sets of tasks. The developed method returns the classification of the signal content, in a window-by-window analysis and can be divided in two distinct phases. The first step consists in the search for a noise free window using a feature obtained from the PCG time-domain. In the second step, the noise free window is compared with the remaining signal. The classification between clean and contaminated PCG is performed using two features from the frequency domain. The algorithm was able to discriminate clean from contamined PCG sections with an average sensitivity and specificity of 95.59% and 92.68%, respectively.

Saccharomyces cerevisiae: a potential host for carboxylic acid production from lignocellulosic feedstock?

Carboxylic acids are important bulk chemicals that can be used as building blocks for the production of polymers, as acidulants, preservatives and flavour compound or as precursors for the synthesis of pharmaceuticals. Today, their production mainly takes place through catalytic processing of petroleum-based precursors. An appealing alternative would be to produce these compounds from renewable resources, using tailor-made microorganisms. Saccharomyces cerevisiae has already demonstrated its value for bioethanol production from renewable resources. In this review, we discuss Saccharomyces cerevisiae engineering potential, current strategies for carboxylic acid production as well as the specific challenges linked to the use of lignocellulosic biomass as carbon source.

Reversible catecholamine-induced cardiomyopathy due to pheochromocytoma: case report.

Pheochromocytoma is a tumor originating from chromaffin tissue. It commonly presents with symptoms and signs of catecholamine excess, such as hypertension, tachycardia, headache and sweating. Cardiovascular manifestations include catecholamine-induced cardiomyopathy, which may present as severe left ventricular dysfunction and congestive heart failure. We report a case of pheochromocytoma which was diagnosed following investigation of dilated cardiomyopathy. We highlight the dramatic symptomatic improvement and reversal of cardiomyopathy, with recovery of left ventricular function after treatment.

Pulmonary hypertension in Portugal: first data from a nationwide registry.

INTRODUCTION: Pulmonary arterial hypertension (PAH) is a rare disease that must be managed in specialized centers; therefore, the availability of epidemiological national data is critical. METHODS: We conducted a prospective, observational, and multicenter registry with a joint collaboration from five centers from Portugal and included adult incident patients with PAH or chronic thromboembolic pulmonary hypertension (CTEPH). RESULTS: Of the 79 patients enrolled in this study, 46 (58.2%) were classified as PAH and 33 patients (41.8%) as CTEPH. PAH patients had a mean age of 43.4 ± 16.4 years. Idiopathic PAH was the most common etiology (37%). At presentation, PAH patients had elevated right atrial pressure (RAP) (7.7 ± 5.9 mmHg) and mean pulmonary vascular resistance (11.4 ± 6.5 Wood units), with a low cardiac index (2.7 ± 1.1 L·min(-1)·m(-2)); no patient was under selective pulmonary vasodilators; however, at follow-up, most patients were on single (50%), double (28%), or triple (9%) combination vasodilator therapy. One-year survival was 93.5%, similar to CTEPH patients (93.9%), that were older (60.0 ± 12.5 years) and had higher RAP (11.0 ± 5.2 mmHg, P = 0.015). CONCLUSIONS: We describe for the first time nationwide data on the diagnosis, management, and prognosis of PAH and CTEPH patients in Portugal. Clinical presentation and outcomes are comparable with those reported on other national registries.