Unlocking bacterial defense: Exploring the potent inhibition of NorA efflux pump by coumarin derivatives in Staphylococcus aureus.

The occurrence of bacterial resistance has been increasing, compromising the treatment of various infections. The high virulence of Staphylococcus aureus allows for the maintenance of the infectious process, causing many deaths and hospitalizations. The MepA and NorA efflux pumps are transporter proteins responsible for expelling antimicrobial agents such as fluoroquinolones from the bacterial cell. Coumarins are phenolic compounds that have been studied for their diverse biological actions, including against bacteria. A pharmacokinetic in silico characterization of compounds C10, C11, C13, and C14 was carried out according to the principles of Lipinski's Rule of Five, in addition to searching for similarity in ChemBL and subsequent search for publications in CAS SciFinder. All compounds were evaluated for their in vitro antibacterial and modulatory activity against standard and multidrug-resistant Gram-positive and Gram-negative strains. The effect of coumarins C9, C10, C11, C13, and C14 as efflux pump inhibitors in Staphylococcus aureus strains was evaluated using the microdilution method (MepA or NorA) and fluorimetry (NorA). The behavior of coumarins regarding the efflux pump was determined from their interaction properties with the membrane and coumarin-protein using molecular docking and molecular dynamics simulations. Only the isolated coumarin compound C13 showed antibacterial activity against standard strains of Staphylococcus aureus and Escherichia coli. However, the other tested coumarins showed modulatory capacity for fluoroquinolone and aminoglycoside antibacterials. Compounds C10, C13, and C14 were effective in reducing the MIC of both antibiotics for both multidrug-resistant strains, while C11 potentiated the effect of norfloxacin and gentamicin for Gram-positive and Gram-negative bacteria and only norfloxacin for Gram-negative. Only coumarin C14 produced synergistic effects when associated with ciprofloxacin in MepA-carrying strains. All tested coumarins have the ability to inhibit the NorA efflux pump present in Staphylococcus aureus, both in reducing the MIC and inducing increased ethidium bromide fluorescence emission in fluorimetry. The findings of this study offer an atomistic perspective on the potential of coumarins as active inhibitors of the NorA pump, highlighting their specific mode of action mainly targeting protein inhibition. In molecular docking, it was observed that coumarins are capable of interacting with various amino acid residues of the NorA pump. The simulation showed that coumarin C10 can cross the bilayer; however, the other coumarins interacted with the membrane but were unable to cross it. Coumarins demonstrated their potentiating role in the effect of norfloxacin through a dual mechanism: efflux pump inhibition through direct interaction with the protein (C9, C10, C11, and C13) and increased interaction with the membrane (C10 and C13). In the context of pharmacokinetic prediction studies, the studied structures have a suitable chemical profile for possible oral use. We suggest that coumarin derivatives may be an interesting alternative in the future for the treatment of resistant bacterial infections, with the possibility of a synergistic effect with other antibacterials, although further studies are needed to characterize their therapeutic effects and toxicity.

Harnessing ultraconfined graphene plasmons to probe the electrodynamics of superconductors.

We show that the Higgs mode of a superconductor, which is usually challenging to observe by far-field optics, can be made clearly visible using near-field optics by harnessing ultraconfined graphene plasmons. As near-field sources we investigate two examples: graphene plasmons and quantum emitters. In both cases the coupling to the Higgs mode is clearly visible. In the case of the graphene plasmons, the coupling is signaled by a clear anticrossing stemming from the interaction of graphene plasmons with the Higgs mode of the superconductor. In the case of the quantum emitters, the Higgs mode is observable through the Purcell effect. When combining the superconductor, graphene, and the quantum emitters, a number of experimental knobs become available for unveiling and studying the electrodynamics of superconductors.

Effects of different temperature-humidity indexes on milk traits of Holstein cows: A 10-year retrospective study.

Test-day records (n = 723,091) collected between 2012 and 2021 from 43,015 Holstein cows at 157 farms located in northern Italy were used to study the effects of heat load on milk production and composition a posteriori. The data consisted of milk yield (kg/d), traditional gross composition traits, somatic cell score (SCS), differential somatic cell count (%), milk ß-hydroxybutyrate (BHB, mmol/L), milk urea (mg/dL), and milk fatty acid composition (g/100 g of milk). Test-day records were then associated with their relative temperature-humidity indexes (THI) calculated using historical environmental data registered by weather stations. Indexes were created using either yearly or summer THI data. The yearly indexes included the average daily THI (adTHI) and the maximum daily THI (mdTHI) measured throughout the whole year, and the summer indexes focused on 3 mo only (June-August) and included the average daily summer THI (adTHIs), the maximum daily summer THI (mdTHIs), and the average daily THI of the hottest 4 h of the day (adTHI4h; 1200-1600 h). All indexes had significant effects on the majority of milk traits analyzed, with, in particular, adTHI and mdTHI being highly significant in explaining the variation of all traits. Milk yield started to decline at a higher THI compared with protein and fat content. The reduction in fat ceased in the elevated THI experienced during the summer months, as demonstrated by adTHIs, mdTHIs, and adTHI4h. The cows had a tendency for increased BHB concentration with elevated THI, suggesting a greater risk of negative energy balance in presence of heat stress. Furthermore, the concentration of the de novo fatty acids C14:0 and C16:0 was reduced in higher THI, reflecting altered mammary gland activity upon elevated heat load and stress. Milk SCS tended to increase with higher adTHIs, mdTHIs, and adTHI4h. The use of yearly indexes is recommended when investigating the effects of heat load on milk composition, whereas summer indexes are suggested when investigating traits influenced by extreme conditions, such as SCS and milk yield. With global temperatures expected to further rise in the upcoming decades, early and easy identification of cows or herds suffering from heat stress, such as through changes in milk composition, is crucial for timely intervention. Adapting measures to mitigate such effects of elevated THI on milk yield and composition is a necessity for the dairy industry to prevent detrimental impacts on dairy production.

Strongly Coupled Magnon-Plasmon Polaritons in Graphene-Two-Dimensional Ferromagnet Heterostructures.

Magnons and plasmons are different collective modes, involving the spin and charge degrees of freedom, respectively. Formation of hybrid plasmon-magnon polaritons in heterostructures of plasmonic and magnetic systems faces two challenges, the small interaction of the electromagnetic field of the plasmon with the spins, and the energy mismatch, as in most systems plasmons have energies orders of magnitude larger than those of magnons. We show that graphene plasmons form polaritons with the magnons of two-dimensional ferromagnetic insulators, placed up to to half a micrometer apart, with Rabi splittings in the range of 100 GHz (dramatically larger than cavity magnonics). This is facilitated both by the small energy of graphene plasmons and the cooperative super-radiant nature of the plasmon-magnon coupling afforded by phase matching. We show that the coupling can be modulated both electrically and mechanically, and we propose a ferromagnetic resonance experiment implemented with a two-dimensional ferromagnet driven by graphene plasmons.

HPV-related oropharyngeal carcinoma remains infrequent over 25 years in a Brazilian Oral Pathology Center: A cross-sectional study with literature review.

BACKGROUND: The aim was to evaluate the frequency, clinicopathological features, and HPV status of oropharyngeal squamous cell carcinoma (OP-SCC) and benign HPV-related epithelial lesions of the oropharynx over the last 25 years. Moreover, a literature review was performed to investigate HPV frequency in OP-SCC samples diagnosed in Brazilian Centers. MATERIAL AND METHODS: A cross-sectional study analyzed OP-SCC, squamous papilloma, verruca vulgaris, and condyloma accuminatum, diagnosed from 1997 to 2021. HPV status of OP-SCC was determined by immunohistochemistry and "in situ" hybridization. Bivariate statistics were performed (p≤0.05). For the literature review, MEDLINE/PubMed, Web of Science, EMBASE, and Scopus were searched. Two independent reviewers assessed the studies for eligibility. RESULTS: Cross-sectional: 211 OP-SCC (63.0%) and 124 benign lesions (37.0%) were included. OP-SCC frequency increased gradually over time, whereas benign lesions had steady trends. OP-SCC affected more males (n= 171; 81.0%), though the relative frequency in females rose over time. Smoking (n= 127; 60.2%) was common in OP-SCC. Nineteen OP-SCC (13.0%) were positive for HPV. HPV-positive and HPV-negative tumors had similar clinicopathological features (p>0.05). Benign lesions predominated in middle-aged (n= 32; 26.7%) women (n= 71; 57.3%), in the soft palate (n=101; 81.5%). LITERATURE REVIEW: 32 studies were included, and in 60% of them, HPV frequency in OP-SCC was less than 25%. CONCLUSIONS: OP-SCC prevalence has been increasing, and it was mostly associated with smoking and alcohol rather than with HPV infection in Brazil. Benign lesions had a stationary frequency over the evaluated period.

Splicing machinery is profoundly altered in systemic lupus erythematosus and antiphospholipid syndrome and directly linked to key clinical features.

OBJECTIVES: To characterize the splicing machinery (SM) of leukocytes from primary antiphospholipid syndrome (APS), systemic lupus erythematosus (SLE) and antiphospholipid syndrome with lupus (APS + SLE) patients, and to assess its clinical involvement. METHODS: Monocytes, lymphocytes and neutrophils from 80 patients (22 APS, 35 SLE and 23 APS + SLE) and 50 HD were purified, and 45 selected SM components were evaluated by qPCR-microfluidic array. Relationship with clinical features and underlying regulatory mechanisms were assessed. RESULTS: APS, SLE and APS + SLE leukocytes displayed significant and specific alterations in SM-components (SMC), associated with clinical features [autoimmune profiles, disease activity, lupus nephritis (LN), and CV-risk markers]. A remarkable relationship among dysregulated SMC in monocytes and the presence of LN in SLE was highlighted, revealing a novel pathological mechanism, which was further explored. Immunohistology analysis of renal biopsies highlighted the pathological role of the myeloid compartment in LN. Transcriptomic analysis of monocytes from SLE-LN(+) vs SLE-LN(-) identified 271 genes differentially expressed, mainly involved in inflammation and IFN-signaling. Levels of IFN-related genes correlated with those of SMC in SLE-LN(+). These results were validated in two external SLE-LN(+) datasets of whole-blood and kidney biopsies. In vitro, SLE-LN(+)-serum promoted a concomitant dysregulation of both, the IFN signature and several SMC, further reversed by JAKinibs treatment. Interestingly, IFNs, key inflammatory cytokines in SLE pathology, also altered SMC. Lastly, the over/down-expression of selected SMC in SLE-monocytes reduced the release of inflammatory cytokines and their adhesion capacity. CONCLUSION: Overall, we have identified, for the first time, a specific alteration of SMC in leukocytes from APS, SLE and APS + SLE patients that would be responsible for the development of distinctive clinical profiles.

Clinical characteristics of 80 subjects with KCNQ2-related encephalopathy: Results from a family-driven survey.

Variants of KCNQ2 are associated with a wide spectrum of disorders, ranging from Self-limiting Neonatal Epilepsy (SelNE) to Early Onset Developmental and Epileptic Encephalopathy (KCNQ2-DEE). Comorbidities associated with this end of the spectrum have been seldomly described and their impact on the life of patients and their families is yet to be investigated. Collaborating with caregivers from different European family associations, we have developed a questionnaire aimed at investigating the onset and frequency of epileptic seizures, anti-seizure medications (ASM), hospitalizations, stages of development, and comorbidities. Responses from 80 patients, 40 males, from 14 countries have been collected. Median age 7.6 years (4 months - 43.6 years). Of 76 epileptic patients (93.6%), 55.3% were seizure-free with a mean age at last seizure of 26.7 months. Among patients with active epilepsy, those older have a lower frequency of seizures (p > 0.05). We were able to identify three different clusters of varying severity (Mild, Severe, Profound), based on neurodevelopmental features and symptoms, excluding epilepsy. Patients in a higher severity cluster had a higher mean number of comorbidities, which had a higher impact on families. Notably, patients in different clusters presented different epilepsy onset and courses. This study constitutes the most extensive data collection of patients with KCNQ2-DEE, with a focus on comorbidities in a wide age group. The participation of caregivers helps to define the impact of the disease on the lives of patients and families and can help identify new primary and secondary outcomes beyond seizures in future studies.

Establishment and cryptic transmission of Zika virus in Brazil and the Americas.

Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.

Minerals and essential amino acids of bovine colostrum: Phenotypic variability and predictive ability of mid- and near-infrared spectroscopy.

Colostrum quality and volume are fundamental for calves because it is the primary supplier of antibodies and the first source of energy, carbohydrates, lipids, proteins, minerals, and vitamins for the newborn. Assessing the detailed composition (i.e., AA and mineral content) of bovine colostrum (BC) on-line and at a reasonable cost would help dairy stakeholders such as farmers or veterinarians for precision feeding purposes and industries producing preparations containing BC such as foodstuff, supplements, and medicaments. In the present study we evaluated mid- (MIRS) and near-infrared spectroscopy (NIRS) prediction ability for AA and mineral composition of individual BC. Second, we the investigated the major factors affecting the phenotypic variability of such traits also evaluating the correlations with the Ig concentration. Results demonstrated that MIRS and NIRS were able to provide sufficiently accurate predictions for all the AA. The coefficient of determination in external validation (R2V) fell, in fact, within the range of 0.70 to 0.86, with the exception of Ile, His, and Met. Only some minerals reached a sufficient accuracy (i.e., Ca, P, S, and Mg; R2V ≥ 0.66) using MIRS, and also S (R2V = 0.87) using NIRS. Phenotypically, both parity and calving season affected the variability of these BC composition traits. Heifers' colostrum was the one with the greatest concentration of Ca and P, the 2 most abundant minerals. These minerals were however very low in cows calving in summer compared with the rest of the year. The pattern of AA across parities and calving season was not linear, likely because their variability was scarcely (or not) affected by these effects. Finally, samples characterized by high IgG concentration were those presenting on average greater concentration of AA. Findings suggest that infrared spectroscopy has the potential to be used to predict certain AA and minerals, outlining the possibility of implementing on-site analyses for the evaluation of the broad-sense BC quality.

Mid-infrared spectroscopy for large-scale phenotyping of bovine colostrum gross composition and immunoglobulin concentration.

Immunoglobulin G is the fundamental antibody for acquisition of passive transfer of immunity in ruminant newborns. Colostrum, in fact, must be administered as soon as possible after birth to ensure a successful transfer of IgG from the dam to the calf. Assessment of colostrum Ig concentration and gross composition via gold standards is expensive, time consuming, and hardly implementable for large-scale investigations. Therefore, in the present study we evaluated the predictive ability of mid-infrared spectroscopy (MIRS) as an indirect determination method. A total of 714 colostrum samples collected within 6 h from parturition from Italian Holstein cows, 30% primiparous and 70% pluriparous, were scanned using a benchtop spectrometer after dilution in pure water. The prediction models were developed by correlating spectral information with the reference measurements: IgG concentration (93.54 ± 33.87 g/L), total Ig concentrations (102.82 ± 35.04 g/L), and content of protein (14.71 ± 3.51%), fat (4.61 ± 3.04%), and lactose (2.36 ± 0.51 mg/100 mg). We found a good to excellent performance in prediction of colostrum IgG concentration and traditional composition traits in cross-validation (R2CV ≥ 0.92) and a promising and good predictive ability in external validation with R2V equal to 0.84, 0.89, and 0.74 for IgG, protein, and fat, respectively. In the case of IgG and protein content, for example, the coefficient of determination in external validation was greater than 0.84. The other Ig fractions, A and M, presented insufficient prediction accuracy likely due to their extremely low concentration compared with IgG (4.56 and 5.06 g/L vs. 93.54 g/L). The discriminant ability of MIRS-predicted IgG and protein content was outstanding when trying to classify samples according to the quality level (i.e., low vs. high concentration of IgG). In particular, the cut-off that better discriminate low- from high-quality colostrum was 75.40 g/L in the case of the MIRS-predicted IgG and 13.32% for the MIRS-predicted protein content. Therefore, MIRS is proposed as a rapid and cheap tool for large-scale punctual IgG, protein, and lactose quantification and for the screening of low-quality samples. From a practical perspective, there is the possibility to install colostrum models in the MIRS benchtop machineries already present in laboratories in charge of official milk testing. Colostrum phenotypes collected on an individual basis will be useful to breeders for the definition of specific selection strategies and to farmers for management scopes. Finally, our findings may be relevant for other stakeholders, given the fact that colostrum is an emerging ingredient for the animal and human food and pharmaceutical industry.

How heat stress conditions affect milk yield, composition, and price in Italian Holstein herds.

An edited data set of 700 bulk and 46,338 test-day records collected between 2019 and 2021 in 42 Holstein-dominated farms in the Veneto Region (North of Italy) was available for the present study. Information on protein, fat and lactose content, somatic cell count, and somatic cell score was available in bulk milk as well as individual test-day records, whereas urea concentration (mg/dL), differential somatic cell count (%), and milk yield (kg/d) were available for test-day records only. Milk features were merged with meteorological data retrieved from 8 weather stations located maximum 10 km from the farms. The daily and weekly temperature-humidity index (THI; wTHI) and maximum daily (MTHI) and weekly temperature-humidity index were associated with each record to evaluate the effect of heat stress conditions on milk-related traits through linear mixed models. Least squares means were estimated to evaluate the effect of THI and, separately, of MTHI on milk characteristics correcting for conventional systematic factors. Overall, heat stress conditions lowered the quality of both bulk milk and test-day records, with fat and protein content being greatly reduced, and somatic cell score and differential somatic cell count augmented. Milk yield was not affected by either THI or MTHI in this data set, but the effect of elevated THI and MTHI was in general stronger on test-day records than on bulk milk. Farm-level economic losses of reduced milk quality rather than reduced yield as consequence of elevated THI or MTHI was estimated to be between $23.57 and $43.98 per farmer per day, which is of comparable magnitude to losses resulting from reduced production. Furthermore, MTHI was found to be a more accurate indicator of heat stress experienced by a cow, explaining more variability of traits compared with THI. The negative effect of heat stress conditions on quality traits commences at lower THI/MTHI values compared with milk yield. Thus, a progressive farmers' income loss due to climatic changes is already a reality and it is mainly due to deterioration of milk quality rather than quantity in the studied area.

Invited review: Iodine level in dairy products-A feed-to-fork overview.

The theme of iodine in the dairy sector is of particular interest due to the involvement and the interconnection of several stakeholders along the dairy food chain. Iodine plays a fundamental role in animal nutrition and physiology, and in cattle it is an essential micronutrient during lactation and for fetal development and the calf's growth. Its correct use in food supplementation is crucial to guarantee the animal's recommended daily requirement to avoid excess intake and long-term toxicity. Milk iodine is fundamental for public health, being one of the major sources of iodine in Mediterranean and Western diets. Public authorities and the scientific community have made great efforts to address how and to what extent different drivers may affect milk iodine concentration. The scientific literature concurs that the amount of iodine administered through animal feed and mineral supplements is the most important factor affecting its concentration in milk of most common dairy species. Additionally, farming practices related to milking (e.g., use of iodized teat sanitizers), herd management (e.g., pasture vs. confinement), and other environmental factors (e.g., seasonality) have been identified as sources of variation of milk iodine concentration. Overall, the aim of this review is to provide a multilevel overview on the mechanisms that contribute to the iodine concentration of milk and dairy products.

Invited review: Bovine colostrum, a promising ingredient for humans and animals-Properties, processing technologies, and uses.

Mammalian colostrum, known as "liquid gold," is considered a valuable source of essential nutrients, growth factors, probiotics, prebiotics, antibodies, and other bioactive compounds. Precisely for this reason, bovine colostrum (BC) is an emerging ingredient for the feed, food, and pharmaceutical industries, being nowadays commercially available in a variety of forms in several countries. Moreover, quite a large number of functional foods and supplements for athletes, human medicines, pet nutrition plans, and complementary feed for some livestock categories, such as piglets and calves, contain BC. The amount of BC yielded by a cow after calving represents approximately 0.5% of the yearly output in dairy breeds. For its nutritional properties and low availability, BC is characterized by a greater market value and an increasing demand compared with other by-products of the dairy sector. However, information regarding the market size of BC for the food and pharmaceutical industries, as well as future developments and perspectives, is scarcely available in the scientific literature. This lack can be attributed to industrial secrecy as well as to the relatively small scale of the BC business when compared with other dairy products, which makes the BC market limited, specific, and intended for a restricted audience. From a legal perspective, regulations assign BC to the large family of milk-derived powders; thus, collecting specific production data, as well as import-export trend information, is not straightforward and can result in unprecise estimates. Given that the interest in BC is increasing in different fields, it is important to have an overview of the production steps and of pros and cons of this emerging ingredient. The present narrative review discloses why BC has started to be considered a product rather than a by-product of the dairy industry. Moreover, the present document aims to summarize the existing methodologies used to assess BC quality in terms of immunoglobulin concentration, the different applications of BC in the industry, and the BC processing technologies. Finally, a panoramic view of the current international market is provided for the first time for this dairy product.

Allergic emergencies in the prehospital setting: a 5 year retrospective study.

Summary: Background. Patients with severe allergic conditions often request support from the prehospital emergency services given the rapid, unexpected and potentially life-threatening nature of the reactions, such as anaphylaxis. Studies regarding prehospital incidents for allergic conditions are scarce. This study aimed to characterize prehospitalar medical requesting assistance due to suspected hypersensitivity reactions (HSR). Methods. Retrospective study of allergic-related requesting assistances between 2017-2022 of a Portuguese emergency dispatch centre - Emergency and Resuscitation Medical Vehicle (VMER), in Coimbra University Hospital. Demographic and clinical variables were analysed, including clinical manifestations, anaphylaxis severity grading, therapeutic interventions, and post-incident allergic work-up. Regarding anaphylactic events, three diagnosis timings were compared: on-site, hospital emergency department and Investigator-diagnosis based on data reviewed. Results. Out of 12689 VMER requesting assistances, 210 (1.7%) were classified as suspected HSR reactions. After on-site medical evaluation, 127 (60.5%) cases maintained the HSR classification (median age 53 years; 56% males) and the main diagnoses included HSR to Hymenoptera venom (29.9%), food allergy (29.1%), and pharmaceutical drugs (25.5%). Anaphylaxis was assumed on-site in 44 (34.7%) cases, in the hospital emergency department in 53 cases (41.7%) and by investigators in 76 (59.8%) cases. Regarding management, epinephrine was administered on-site in 50 cases (39.4%). Conclusions. The main reason for prehospital requesting assistance was HSR to Hymenoptera venom. A high proportion of incidents met the criteria for anaphylaxis and despite the inherent difficulties of the prehospital setting, many of the on-site diagnoses agreed with the criteria. Regarding management, epinephrine was underused in this setting. Referral to specialized consultation is crucial for the management of prehospital incidents.

The stromal immunoexpression of CLIC4 may be related to the difference in the biological behavior between oral squamous cell carcinoma and oral verrucous carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) has high morbidity and mortality rates while oral verrucous carcinoma (OVC), an uncommon variant of OSCC, exhibits a distinct biological behavior. CLIC4 protein plays a role in the cell cycle and apoptosis regulation and participates in the myofibroblasts transdifferentiation process, which are the main cells of the tumor stroma. This study analyzed the immunoexpression of CLIC4 and α-SMA in 20 OSCC cases and 15 OVC cases. MATERIAL AND METHODS: A semiquantitative analysis of CLIC4 and α-SMA immunoexpression was performed in the parenchyma and stroma. Nuclear and cytoplasmic reactivity was analyzed separately for the CLIC4 immunostaining. The data were submitted to Pearson's chi-square and Spearman's correlation tests (p ≤ 0.05). RESULTS: In the CLIC4 analysis, there was a significant difference in the immunoexpression of this protein between OSCC and OVC stroma (p < 0.001). It was observed a higher expression of α-SMA in the OSCC stroma. There was a positive and significant correlation between CLIC4 and α-SMA immunoexpression in the OVC stroma (r = 0,612; p = 0,015). CONCLUSIONS: The decrease or absence of nuclear CLIC4 immunoexpression in the neoplastic epithelial cells and the increase of its expression in the stroma may influence the difference in biological behavior between OSCC and OVC.

Acute rapamycin rescues the hyperexcitable phenotype of accumbal medium spiny neurons in the valproic acid rat model of autism spectrum disorder.

We previously demonstrated that prenatal exposure to valproic acid (VPA), an environmental model of autism spectrum disorder (ASD), leads to a hyperexcitable phenotype associated with downregulation of inward-rectifying potassium currents in nucleus accumbens (NAc) medium spiny neurons (MSNs) of adolescent rats. Aberrant mTOR pathway function has been associated with autistic-like phenotypes in multiple animal models, including gestational exposure to VPA. The purpose of this work was to probe the involvement of the mTOR pathway in VPA-induced alterations of striatal excitability. Adolescent male Wistar rats prenatally exposed to VPA were treated acutely with the mTOR inhibitor rapamycin and used for behavioral tests, ex vivo brain slice electrophysiology, single-neuron morphometric analysis, synaptic protein quantification and gene expression analysis in the NAc. We report that postnatal rapamycin ameliorates the social deficit and reverts the abnormal excitability, but not the inward-rectifying potassium current defect, of accumbal MSNs. Synaptic transmission and neuronal morphology were largely unaffected by prenatal VPA exposure or postnatal rapamycin treatment. Transcriptome analysis revealed extensive deregulation of genes implied in neurodevelopmental disorders and ionic mechanisms exerted by prenatal VPA, which was partially reverted by postnatal rapamycin. The results of this work support the existence of antagonistic interaction between mTOR and VPA-induced pathways on social behavior, neurophysiological phenotype and gene expression profile, thus prompting further investigation of the mTOR pathway in the quest for specific therapeutic targets in ASD.

COVID-19 and HIV/AIDS in a cohort study in Sao Paulo, Brazil: outcomes and disparities by race and schooling.

Studies describing characteristics and outcomes of COVID-19 among people living with HIV are currently limited, lacking detailed evaluation of the interplay among demographics, HIV-related variables, and comorbidities on COVID-19 outcomes. This retrospective cohort study describes mortality rates overall and according to demographic characteristics and explores predictors of admission to intensive care unit and death among 255 persons living with HIV with severe acute respiratory syndrome and confirmed SARS-CoV-2 infection in the State of Sao Paulo, Brazil. We found that the overall mortality rate was 4.1/1,000 person-days, with a case-fatality of 34%. Higher rates occurred among older adults, Black/Mixed skin color/race patients, and those with lower schooling. In a multivariable analysis adjusted for age, sex, CD4 count, viral load and number of comorbidities, skin color/race, and schooling remained significantly associated with higher mortality. Although tenofovir use was more frequent among survivors in the univariable analysis, we failed to find a statistically significant association between tenofovir use and survival in the multivariable analysis. Our findings suggest that social vulnerabilities related to both HIV and COVID-19 significantly impact the risk of death, overtaking traditional risk factors such as age, sex, CD4 count, and comorbidities.

Machine learning for predicting chronic diseases: a systematic review.

OBJECTIVES: We aimed to review the literature regarding the use of machine learning to predict chronic diseases. STUDY DESIGN: This was a systematic review. METHODS: The searches included five databases. We included studies that evaluated the prediction of chronic diseases using machine learning models and reported the area under the receiver operating characteristic curve values. The Transparent Reporting of a multivariable prediction model for Individual Prognosis Or Diagnosis scale was used to assess the quality of studies. RESULTS: In total, 42 studies were selected. The best reported area under the receiver operating characteristic curve value was 1, whereas the worst was 0.74. K-nearest neighbors, Naive Bayes, deep neural networks, and random forest were the machine learning models most frequently used for achieving the best performance. CONCLUSION: We found that machine learning can predict the occurrence of individual chronic diseases, progression, and their determinants and in many contexts. The findings are original and relevant to improve clinical decisions and the organization of health care facilities.

Chlorella vulgaris meets TiO2 NPs: Effective sorbent/photocatalytic hybrid materials for water treatment application.

A new class of bio-nano hybrid catalyst useable in downstream wastewater treatment was developed. We combined the sorption potentialities of Chlorella vulgaris microalgae with the photocatalytic properties of TiO2 NPs in order to investigate unexplored synergistic effects that could push the algal remediation technology toward a more promising cost-effective balance. We exploited non-living C. vulgaris, which keeps the biosorption properties of the living microalgae, but greatly enhancing the overall processability. C. vulgaris biomass was coupled with TiO2 NPs and the nanosols were then dried by means of a spray freeze drying (SFD) process able to produce highly reactive granules. A widespread physicochemical characterization supported the preparation and the performance evaluation, so highlighting the key-role of C. vulgaris/TiO2 interaction at the colloidal state. Heavy metal adsorption, tested for copper ions, and photocatalytic activity, assessed for Rhodamine B (RhB) photodegradation, were evaluated as key performances. The results pointed out a positive synergistic effect for hybrid samples consistent with the enhancement of metal biosorption which ranges from 103 mg g-1, for pristine C. vulgaris, to about 4000 mg g-1, when the biomass was coupled with the inorganic nanophase. The photocatalytic activity was well preserved with a complete RhB conversion after 1 h and even advanced in presence of SiO2NPs into the inorganic counterpart, so increasing the kinetic constant from 8.70 to 10.7 10-2 min-1. The results pave the way for the integration of these sorbent/photocatalytic hybrid materials into water remediation systems in an innovative sustainable design perspective.

Prognostic and clinicopathological significance of tumor-stroma ratio in head and neck squamous cell carcinoma: A systematic review.

BACKGROUND: Analysis of the tumor microenvironment has been proposed as a strategy for the treatment and prognosis of different neoplastic processes. A grading system based on the tumor-stroma ratio (TSR), which evaluates the proportion of stroma in relation to neoplastic parenchyma at the invasion front, has shown a strong prognostic value in different neoplastic processes. The aim of the present systematic review was to understand the role of the TSR in head and neck squamous cell carcinoma (HNSCC), evaluating its correlation with clinical and prognostic parameters. MATERIAL AND METHODS: An electronic search was performed in PubMed/Medline, Web of Science, Science Direct, Scopus, Embase, and the Cochrane Collaboration Library. Publications assessing the relationship between TSR and prognosis in cases of HNSCC were eligible. The quality of the studies was assessed independently by four evaluators using the Newcastle-Ottawa scale. RESULTS: After application of the previously es+lished inclusion/exclusion criteria, nine articles were included in the qualitative synthesis. With regards to quality on the Newcastle-Ottawa scale, an overall value of 4.55 was obtained. This systematic review demonstrated a strong association between TSR and prognosis in esophageal and oral squamous cell carcinomas. CONCLUSIONS: Histopathological analysis of the TSR can optimize the analysis of the prognosis of cases diagnosed with HNSSC. In addition, the TSR is a reliable and simple parameter that can be evaluated in hematoxylin/eosin-stained slides during routine laboratory examinations, showing high inter- and intraobserver agreement.