GABAergic system and chloride cotransporters as potential therapeutic targets to mitigate cell death in ischemia.

Gamma aminobutyric acid (GABA) is a critical inhibitory neurotransmitter in the central nervous system that plays a vital role in modulating neuronal excitability. Dysregulation of GABAergic signaling, particularly involving the cotransporters NKCC1 and KCC2, has been implicated in various pathologies, including epilepsy, schizophrenia, autism spectrum disorder, Down syndrome, and ischemia. NKCC1 facilitates chloride influx, whereas KCC2 mediates chloride efflux via potassium gradient. Altered expression and function of these cotransporters have been associated with excitotoxicity, inflammation, and cellular death in ischemic events characterized by reduced cerebral blood flow, leading to compromised tissue metabolism and subsequent cell death. NKCC1 inhibition has emerged as a potential therapeutic approach to attenuate intracellular chloride accumulation and mitigate neuronal damage during ischemic events. Similarly, targeting KCC2, which regulates chloride efflux, holds promise for improving outcomes and reducing neuronal damage under ischemic conditions. This review emphasizes the critical roles of GABA, NKCC1, and KCC2 in ischemic pathologies and their potential as therapeutic targets. Inhibiting or modulating the activity of these cotransporters represents a promising strategy for reducing neuronal damage, preventing excitotoxicity, and improving neurological outcomes following ischemic events. Furthermore, exploring the interactions between natural compounds and NKCC1/KCC2 provides additional avenues for potential therapeutic interventions for ischemic injury.

Acceleration and transport of relativistic electrons in the jets of the microquasar SS 433.

SS 433 is a microquasar, a stellar binary system that launches collimated relativistic jets. We observed SS 433 in gamma rays using the High Energy Stereoscopic System (H.E.S.S.) and found an energy-dependent shift in the apparent position of the gamma-ray emission from the parsec-scale jets. These observations trace the energetic electron population and indicate that inverse Compton scattering is the emission mechanism of the gamma rays. Our modeling of the energy-dependent gamma-ray morphology constrains the location of particle acceleration and requires an abrupt deceleration of the jet flow. We infer the presence of shocks on either side of the binary system, at distances of 25 to 30 parsecs, and that self-collimation of the precessing jets forms the shocks, which then efficiently accelerate electrons.

Antimicrobial resistance profile of Aeromonas spp. isolated from asymptomatic Colossoma macropomum cultured in the Amazonas State, Brazil.

Bacterial diseases are important factors that limit productivity in aquaculture. To reduce negative economic impacts, fish farmers use antimicrobials, often indiscriminately, and this action has led to bacterial resistance to drugs. The objectives of this study were to isolate and identify the main putative pathogenic bacterial species in tambaqui (Colossoma macropomum), establish the profile of resistance to antimicrobials by the methods of disc diffusion, and determine the minimum inhibitory concentration (MIC) values. Two hundred and ninety asymptomatic fish were collected between March and November 2015 from ten fish farms in the Amazonas state (Brazil). Of the total strains recovered from tambaqui, seven were identified as Aeromonas spp. by sequencing the 16S rRNA gene. These seven isolates showed resistance to ampicillin, 28% to erythromycin, and 28% to sulfonamide. Additionally, the seven isolates showed a MIC higher than the range evaluated for amoxicillin, penicillin, novobiocin, tylosin tartrate, and clindamycin, and 85% showed resistance to erythromycin. The results of this study indicate the need to increase the awareness of fish farmers and, most importantly, the government, about the lack of drug regulations for use in aquaculture, and good management practices, so the indiscriminate prophylactic and systemic use of antimicrobials be inhibited.

Reliability and validity of the Chester step test in patients with interstitial lung disease.

BACKGROUND: The Chester Step Test (CST) is a simple and inexpensive field test, which requires minimal physical space to assess exercise capacity. Such characteristics make the CST suitable to be used in different settings, however, its measurement properties in patients with interstitial lung diseases (ILD) are unknown. METHODS: A cross-sectional study was conducted in patients with ILD. First, a CST-1 and a 6-minute walk test (6MWT) were performed. After 48-72 hours, a CST-2 was repeated. A 2nd rater was present in one of the sessions. Relative reliability was measured using intraclass correlation coefficient (ICC1,1 and ICC2,1). Absolute reliability was determined using standard error of measurement (SEM), minimal detectable change at 95% confidence interval (MDC95) and the Bland-Altman method. The values of SEM and MDC95 were also expressed as a percentage of the mean. Construct validity was explored using Spearman correlation coefficient (rs) between the number of steps taken in the best CST and the distance performed in the 6MWT. RESULTS: Sixty-six patients with ILD (65.5±12.9 years; 48.5%men; FVC 79.4±18.8pp; DLCO 49.0±18.3pp) participated in the study. Relative (ICC 0.95-1.0) and absolute reliability were excellent without evidence of systematic bias. The SEM and MDC95 were 11.8 (14.7%) and 32.6 steps (40.7%), respectively. The correlation between CST and 6MWT was significant, positive, and high (rs=0.85, p=0.001). CONCLUSION: The CST is a reliable and valid test and might be especially useful to assess exercise capacity in patients with ILD in limited space environments.

Search for Dark Matter Annihilation Signals in the H.E.S.S. Inner Galaxy Survey.

The central region of the Milky Way is one of the foremost locations to look for dark matter (DM) signatures. We report the first results on a search for DM particle annihilation signals using new observations from an unprecedented γ-ray survey of the Galactic Center (GC) region, i.e., the Inner Galaxy Survey, at very high energies (≳100 GeV) performed with the H.E.S.S. array of five ground-based Cherenkov telescopes. No significant γ-ray excess is found in the search region of the 2014-2020 dataset and a profile likelihood ratio analysis is carried out to set exclusion limits on the annihilation cross section ⟨σv⟩. Assuming Einasto and Navarro-Frenk-White (NFW) DM density profiles at the GC, these constraints are the strongest obtained so far in the TeV DM mass range. For the Einasto profile, the constraints reach ⟨σv⟩ values of 3.7×10^{-26} cm^{3} s^{-1} for 1.5 TeV DM mass in the W^{+}W^{-} annihilation channel, and 1.2×10^{-26} cm^{3} s^{-1} for 0.7 TeV DM mass in the τ^{+}τ^{-} annihilation channel. With the H.E.S.S. Inner Galaxy Survey, ground-based γ-ray observations thus probe ⟨σv⟩ values expected from thermal-relic annihilating TeV DM particles.

Seed priming with light quality and Cyperus rotundus L. extract modulate the germination and initial growth of Moringa oleifera Lam. seedlings.

Improving plant germination is essential to guarantee better quality seedlings. Thus, this research aimed to evaluate whether the seed priming with light quality (LIQ) and the aqueous extract of Cyperus rotundus (AEC) tuber could modulate the germination and initial growth of Moringa oleifera L. seedlings. The experimental design was a completely randomized in the 4x4 factorial scheme, composed of four LIQ conditions (white, blue, red, and distant red light) and four AEC concentrations (0, 25, 50 and 100%). Seed priming with red light reduced the average emergence time, while blue, red, and extreme red lights associated with 50% of aqueous extract of C. rotundus increased shoot initial length and photosynthetic pigment accumulation. Seed priming with blue light resulted in seedlings with a shorter final shoot length. However, application of 100% of aqueous extract of C. rotundus reversed this. The white light in combination with concentrations of 50 and 100% of AEC promoted a higher relative shoot growth rate of seedlings. The research revealed that seed priming with light quality and aqueous extracts of C. rotundus tubers modulates the germination and initial growth of M. oleifera seedlings. More work needs to be done to determine the responsible compounds in AEC that is responsible for priming growth as phytohormones.

Time-resolved hadronic particle acceleration in the recurrent nova RS Ophiuchi.

Recurrent novae are repeating thermonuclear explosions in the outer layers of white dwarfs, due to the accretion of fresh material from a binary companion. The shock generated when ejected material slams into the companion star's wind can accelerate particles. We report very-high-energy (VHE; [Formula: see text]) gamma rays from the recurrent nova RS Ophiuchi, up to 1 month after its 2021 outburst, observed using the High Energy Stereoscopic System (H.E.S.S.). The temporal profile of VHE emission is similar to that of lower-energy giga-electron volt emission, indicating a common origin, with a 2-day delay in peak flux. These observations constrain models of time-dependent particle energization, favoring a hadronic emission scenario over the leptonic alternative. Shocks in dense winds provide favorable environments for efficient acceleration of cosmic rays to very high energies.

Does the addition of chlorhexidine to glass ionomer cements influence its antimicrobial effect and survival rate? A systematic review.

PURPOSE: To evaluate the influence of the addition of chlorhexidine on the antimicrobial effect and on the survival of restorations performed with glass ionomer cement. METHODS: Nine databases were used to search for randomized clinical trials that compared the survival rate and the antimicrobial effect of glass ionomer cement (GIC) restorations with and without the incorporation of chlorhexidine (CHX), without restrictions on year or language. Cochrane Collaboration's Risk of Bias 2 was used to assess the risk of bias. The GRADE approach was used to assess the certainty of evidence. RESULTS: From 593 studies found, seven met the inclusion criteria. The concentration of CHX varied between 0.5 and 2%. In general, the addition of CHX to GIC promoted reductions in Streptococcus mutans and Lactobacillus acidophilus burdens when compared to those without CHX. No study showed a difference in the survival of restorations between GIC with CHX and conventional GIC. Individual risk of bias varied from low to high and the certainty of evidence was classified as very low. CONCLUSIONS: Based on a very low level of certainty, the evidence suggests that the incorporation of CHX in GIC might improve the antimicrobial effects for a short time, in addition to having little influence on the survival of the restoration.

Assessment of children's exposure to carbonaceous matter and to PM major and trace elements.

Particulate matter (PM) pollution is one of the major environmental concerns due to its harmful effects on human health. As children are particularly vulnerable to particle exposure, this study integrates the concentration of PM chemical compounds measured in the micro-environments (MEs) where children spend most of their time to assess the daily exposure and inhaled dose. PM samples were analysed for organic and elemental carbon and for major and trace elements. Results showed that the MEs that contribute most to the children's daily exposure (80%) and inhaled dose (65%) were homes and schools. Results indicated that the high contribution of particulate organic matter (POM) indoors indicate high contributions of indoor sources to the organic fraction of the particles. The highest concentrations of PM chemical compounds and the highest Indoor/Outdoor ratios were measured in schools, where the contribution of mineral elements stands out due to the resuspension of dust caused by the students and to the chalk used in blackboards. The contribution of the outdoor particles to inhaled dose (24%) was higher than to the exposure (12%), due to the highest inhalation rates associated with the activities performed outdoor. This study indicates the importance of indoor air quality for the children's exposure and health.

Revealing x-ray and gamma ray temporal and spectral similarities in the GRB 190829A afterglow.

Gamma-ray bursts (GRBs), which are bright flashes of gamma rays from extragalactic sources followed by fading afterglow emission, are associated with stellar core collapse events. We report the detection of very-high-energy (VHE) gamma rays from the afterglow of GRB 190829A, between 4 and 56 hours after the trigger, using the High Energy Stereoscopic System (H.E.S.S.). The low luminosity and redshift of GRB 190829A reduce both internal and external absorption, allowing determination of its intrinsic energy spectrum. Between energies of 0.18 and 3.3 tera-electron volts, this spectrum is described by a power law with photon index of 2.07 ± 0.09, similar to the x-ray spectrum. The x-ray and VHE gamma-ray light curves also show similar decay profiles. These similar characteristics in the x-ray and gamma-ray bands challenge GRB afterglow emission scenarios.

Forced degradation study and characterization of main impurities of ibuprofen soft gelatin capsules by LC-MS-QTOF.

Ibuprofen soft gelatin capsules were subjected to degradation under acidic, basic, oxidation, photolytic, thermal, humidity, and metal ions conditions. To analyse the degradation products, a reversed-phase liquid chromatography (RP-LC) indicative stability method was successfully developed. Chromatographic separation was achieved using a Poroshell HPH-C18 150 x 4.6 mm, 4 µm, column at 25 °C, with a mobile phase constituted by 0.1% phosphoric acid and acetonitrile in gradient at a flow rate of 1.0 mL• min -1 , using ultraviolet detection at 220 nm and injection volume of 20 µL. In total, eight unknown impurities were found. The peaks RRt 0.49, RRt 0.75, and RRt 0.95 were above 0.17%, corresponding to the identification threshold. Those were identified and characterized by LC-MS-QTOF, with the same chromatographic conditions, except for the exchange of 0.1% phosphoric acid for 0.1% formic acid. The impurities originated from the interaction of ibuprofen with excipients: esterification with PEG, with sorbitol/sorbitan, and with glycerol by-products, which has not yet been reported in the literature. The developed method can be used in several pharmaceutical areas as quality control of impurities, studies of forced degradation, and for the development of future formulations.

Organophosphate esters in airborne particles from subway stations.

For the first time, the concentrations of 19 organophosphate esters (OPEs) were measured in airborne fine particulate matter (PM2.5) from subway stations in Barcelona (Spain) to investigate their occurrence, contamination profiles and associated health risks. OPEs were detected in all PM2.5 samples with levels ranging between 1.59 and 202 ng/m3 (mean value of 39.9 ng/m3). Seventeen out of 19 tested analytes were detected, with TDClPP, TClPP and TCEP being those presenting the highest concentrations. OPE concentrations are not driven by the same factors that determine the ambient PM2.5 concentrations of other constituents in the subway. Newer stations presented higher OPE levels, probably due to the materials used in the design of the platforms, with greater use of modern plastic materials versus older stations with tiles and stones. Estimated daily intakes via airborne particles inhalation during the time expended in subway stations were calculated, as well as the carcinogenic and non-carcinogenic health risks (CR and non-CR), all being much lower than the threshold risk values. Thus, subway inhalation exposure when standing on the platform to OPE's per se is not considered to be dangerous for commuters.

MT1 and MT2 melatonin receptors play opposite roles in brain cancer progression.

Primary brain tumors remain among the deadliest of all cancers. Glioma grade IV (glioblastoma), the most common and malignant type of brain cancer, is associated with a 5-year survival rate of < 5%. Melatonin has been widely reported as an anticancer molecule, and we have recently demonstrated that the ability of gliomas to synthesize and accumulate this indolamine in the surrounding microenvironment negatively correlates with tumor malignancy. However, our understanding of the specific effects mediated through the activation of melatonin membrane receptors remains limited. Thus, here we investigated the specific roles of MT1 and MT2 in gliomas and medulloblastomas. Using the MT2 antagonist DH97, we showed that MT1 activation has a negative impact on the proliferation of human glioma and medulloblastoma cell lines, while MT2 activation has an opposite effect. Accordingly, gliomas have a decreased mRNA expression of MT1 (also known as MTNR1A) and an increased mRNA expression of MT2 (also known as MTNR1B) compared to the normal brain cortex. The MT1/MT2 expression ratio negatively correlates with the expression of cell cycle-related genes and is a positive prognostic factor in gliomas. Notably, we showed that functional selective drugs that simultaneously activate MT1 and inhibit MT2 exert robust anti-tumor effects in vitro and in vivo, downregulating the expression of cell cycle and energy metabolism genes in glioma stem-like cells. Overall, we provided the first evidence regarding the differential roles of MT1 and MT2 in brain tumor progression, highlighting their relevance as druggable targets. KEY MESSAGES: • MT1 impairs while MT2 promotes the proliferation of glioma and medulloblastoma cell lines. • Gliomas have a decreased expression of MT1 and an increased expression of MT2 compared to normal brain cortex. • Tumors with a high MT1/MT2 expression ratio have significantly better survival rates. • Functional selective drugs that simultaneously activate MT1 and inhibit MT2 downregulate the expression of cell cycle and energy metabolism genes in glioma stem-like cells and exert robust anti-tumor effects in vivo.

Loss of the laminin subunit alpha-3 induces cell invasion and macrophage infiltration in cutaneous squamous cell carcinoma.

BACKGROUND: Cutaneous squamous cell carcinoma (cSCC) is a common cancer that invades the dermis through the basement membrane. The role of the basement membrane in poorly differentiated cSCC is not well understood. OBJECTIVES: To study the effect that loss of the laminin subunit alpha-3 (α3) chain from the tumour microenvironment has on tumour invasion and inflammatory cell recruitment. METHODS: We examined the role of the basement membrane proteins laminin subunits α3, ß3 and γ2 in SCC invasion and inflammatory cell recruitment using immunohistochemistry, short hairpin RNA knockdown, RNA-Seq, mouse xenograft models and patient tumour samples. RESULTS: Analysis of SCC tumours and cell lines using antibodies specific to laminin chains α3, ß3 and γ2 identified a link between poorly differentiated SCC and reduced expression of laminin α3 but not the other laminin subunits investigated. Knockdown of laminin α3 increased tumour invasion both in vitro and in vivo. Western blot and immunohistochemical staining identified increased phosphorylated myosin light chain with loss of laminin α3. Inhibition of ROCK (rho-associated protein kinase) but not Rac1 significantly reduced the invasive potential of laminin α3 knockdown cells. Knockdown of laminin subunits α3 and γ2 increased monocyte recruitment to the tumour microenvironment. However, only the loss of laminin α3 correlated with increased tumour-associated macrophages both in xenografted tumours and in patient tumour samples. CONCLUSIONS: These data provide evidence that loss of the laminin α3 chain in cSCC has an effect on both the epithelial and immune components of cSCC, resulting in an aggressive tumour microenvironment.

Micellar effects and analytical applications of nitro substitution in 4-Nitro-N-alkyl-1,8-naphthalimide by cysteine derivatives.

The aromatic nucleophilic substitution reactions of the nitro group of 4-Nitro-N-alkyl-1,8-naphthalimides by thiolate anions produce fluorescent derivatives and their rates are strongly accelerated by micelles of hexadecyltrimethylammonium chloride even at low pH. Acceleration factors of this reactions can reach million-fold. As the products are oxidant-insensible, this reaction allows the determination of SH- containing compounds such as cysteine, glutathione or proteins even in oxidative conditions. Limits of detection are as low as 5 × 10-7 M, ten times lower than the limit for the classic 5,5'-dithiobis-(2-nitrobenzoic) acid method. Moreover, this reaction can be developed at pHs between 6.5 and 7.5 thereby diminishing the rate of spontaneous oxidation of the thiols. In addition, we demonstrated that 4-Nitro-N-alkyl-1,8-naphthalimides can be used to evidence SH groups in peptides, proteins and living cells.

Antioxidant effect on viability of boar semen cooled to 15°C / Influência de antioxidantes na viabilidade do sêmen suíno resfriado a 15°C

This study aimed to evaluate the addition of Vitamin C, reduced Glutathione and trolox on sperm characteristics of pork refrigerated semen. Six pigs were collected through the technique of gloved hand (10 ejaculates/animals). The semen was diluted in MR-A®. After the previous evaluations, the treatments were added: Control group: diluent only; Vitamin C Group: 200µM/mL Vitamin C; Trolox Group: 200µM/mL Trolox; Glutathione group: 2.5mM/ml Reduced glutathione. The semen was stored in thermal boxes and placed inside the refrigerator at 15oC and evaluated at D0, 12, 48, 72 hours. After 30 hours of incubation, each treatment was divided into two equal fractions and the same concentration of antioxidants was added in one of the parts. The results show that reduced glutathione supplementation preserves sperm motility after 24 hours but also has a higher percentage of acrosome intact in the presence of this antioxidant. There was no effect of adding a second dose of the antioxidants. In conclusion, the addition of reduced Glutathione to the swine semen diluent is a promising alternative for better preservation of sperm characteristics and the addition of the second dose of antioxidants during storage is detrimental to semen.(AU)

Este estudo tem como objetivo avaliar a adição da vitamina C, da glutationa reduzida e do trolox sobre características espermáticas do sêmen refrigerado de suínos. Seis cachaços foram coletados pela técnica de mão enluvada (10 coletas/animal). O sêmen foi diluído em MR-A®. Após as avaliações prévias, os tratamentos foram adicionados: grupo controle: apenas diluidor; grupo vitamina C: 200µM/mL de vitamina C; grupo trolox: 200µM/mL de trolox; grupo glutationa: 2.5mM/mL de glutationa reduzida. O sêmen foi armazenado em caixas térmicas e alocado dentro do refrigerador a 15oC e avaliado nos tempos zero, 12, 48 e 72 horas . Após 30 horas de incubação, cada tratamento foi dividido em duas frações iguais e adicionou-se a mesma concentração de antioxidantes em uma das partes. Os resultados demonstram que a suplementação de glutationa reduzida preserva a motilidade espermática após 24 horas, bem como tem maior percentual de acrossoma intacto na presença desse antioxidante. Não houve efeito ao se adicionar uma segunda dose dos antioxidantes. Em conclusão, o acréscimo da glutationa reduzida ao diluidor de sêmen suíno é uma alternativa promissora para melhor preservação das características espermáticas, e a adição da segunda dose dos antioxidantes durante o armazenamento é prejudicial ao sêmen.(AU)

Predictors of nutritional and inflammation risk in hemodialysis patients.

BACKGROUND: Malnutrition and chronic inflammation are prevalent complications in hemodialysis (HD) patients. Different nutritional assessment tools are used to identify patients at risk. A composite and comprehensive malnutrition inflammation score (MIS) has been correlated with morbidity and mortality, and appears to be a robust and quantitative tool. OBJECTIVES: Determine malnutrition risk profile in a sample of portuguese HD patients; determine the association of clinical and laboratory factors with MIS, and the impact of each parameter on MIS. METHODS AND RESULTS: We performed, between September 15th of 2015 and January 31st of 2016, a cross sectional analysis of 2975 patients, representing 25% of portuguese HD patients. 59% were men (66.7 ± 14.8 years); 31% diabetic; 79% and 21% performed, respectively, high-flux HD and HDF. A MIS >5 was considered to indicate higher risk and was present in 1489 patients (50%). Amongst all parameters, comorbilities/dialysis vintage, transferrin, functional capacity, changes in body weight and decreased fat stores showed the higher impact, while albumin had one of the lowest impact on the nutritional risk. MULTIVARIABLE ANALYSIS: Higher age (>75 years, OR 1.71, p < 0.001), diabetes (OR 1.25, p = 0.026), lower P levels (OR 1.57,p = 0.001), higher Ca levels (OR 1.51, p < 0.001), higher ERI (OR 1.05, p < 0.001), higher Kt/V (OR 2.14, p < 0.001) and higher CRP (OR 1.01, p < 0.001) were independently associated with a higher risk of MIS>5; higher nPNA (OR 0.29, p < 0.001) and higher Pcreat (OR 0.88, p < 0.001) were associated with a risk reduction of MIS>5 (95% CI). CONCLUSIONS: Routine clinical and analytic parameters were found to be associated with MIS range that might indicate higher risk, and may represent a simple alert sign for the need of further assessments.

Particle exposure and inhaled dose while commuting in Lisbon.

While commuting, individuals are exposed to high concentrations of urban air pollutants that can lead to adverse health effects. This study aims to assess commuters' exposure to particulate matter (PM) when travelling by car, bicycle, metro and bus in Lisbon. Mass concentrations of PM2.5 and PM10 were higher in the metro. On the other hand, the highest BC and PN0.01-1 average concentrations were found in car and bus mode, respectively. In cars, the outdoor concentrations and the type of ventilation appeared to affect the indoor concentrations. In fact, the use of ventilation led to a decrease of PM2.5 and PM10 concentrations and to an increase of BC concentrations. The highest inhaled doses were mostly observed in bicycle journeys, due to the longest travel periods combined with enhanced physical activity and, consequently, highest inhalation rates.