Enhanced Assessment of Cross-Reactive Antigenic Determinants within the Spike Protein.

Despite successful vaccination efforts, the emergence of new SARS-CoV-2 variants poses ongoing challenges to control COVID-19. Understanding humoral responses regarding SARS-CoV-2 infections and their impact is crucial for developing future vaccines that are effective worldwide. Here, we identified 41 immunodominant linear B-cell epitopes in its spike glycoprotein with an SPOT synthesis peptide array probed with a pool of serum from hospitalized COVID-19 patients. The bioinformatics showed a restricted set of epitopes unique to SARS-CoV-2 compared to other coronavirus family members. Potential crosstalk was also detected with Dengue virus (DENV), which was confirmed by screening individuals infected with DENV before the COVID-19 pandemic in a commercial ELISA for anti-SARS-CoV-2 antibodies. A high-resolution evaluation of antibody reactivity against peptides representing epitopes in the spike protein identified ten sequences in the NTD, RBD, and S2 domains. Functionally, antibody-dependent enhancement (ADE) in SARS-CoV-2 infections of monocytes was observed in vitro with pre-pandemic Dengue-positive sera. A significant increase in viral load was measured compared to that of the controls, with no detectable neutralization or considerable cell death, suggesting its role in viral entry. Cross-reactivity against peptides from spike proteins was observed for the pre-pandemic sera. This study highlights the importance of identifying specific epitopes generated during the humoral response to a pathogenic infection to understand the potential interplay of previous and future infections on diseases and their impact on vaccinations and immunodiagnostics.

"RAIL BIOPSY" A Novel and Useful Technique for Hysteroscopic Endometrial Target Biopsy.

STUDY OBJECTIVE: Endometrial biopsy (EB) is one of the most common gynecologic procedures. Office-based EB has replaced procedures involving general/loco-regional anesthesia and cervical dilatation performed in the operating room [1-3]. The Grasp Biopsy seems to be the most appropriate EB technique for reproductive-aged women [1,2,4]. Recently, the Visual D&C performed with hysteroscopic tissue removal devices has shown to be a valid alternative [5]. However, it is often difficult to obtain an adequate specimens in peri/post-menopausal women with hypo/atrophic endometrium [2]. Our aim is to show a novel hysteroscopic EB technique called "Rail Biopsy" which requires widespread and cheap instruments. DESIGN: A step-by-step explanation of surgical techinque with narrated video footage. SETTING: Tertiary Level Academic Hospital "IRCCS Azienda Ospedaliero-Universitaria di Bologna" Bologna, Italy. INTERVENTIONS: We performed the "Rail Biopsy" technique with a 5.0 mm Continuous Flow Operative Hysteroscope with a 30° Lens and a 5Fr operative channel. We identify the endometrial target area (ETA), and we create a first track cutting through the endometrium in a caudo-cranial direction using cold scissors. We repeat the procedure, creating a second parallel track, thus completing our "rail" and isolating a wide ETA. Then, in the caudo-cranial direction, we cut through the stromal layer beneath the ETA. With a 5Fr cold grasping forceps, we clench the cranial edge of the ETA, and we remove it from the uterine cavity. A high-quality specimen, even in the case of hypo/atrophic endometrium or focal sessile lesions, can be obtained with this technique. The crucial aspect of the "Rail Biopsy" indeed is cutting through the stromal tissue while the endometrium is minimally touched, avoiding thermal damage deriving from electrosurgery. The instruments required are widespread and cheap. Moreover, this technique can be performed on any wall of the uterus, under vision, and, in the majority of patients, in an office-setting without cervical dilatation or general/loco-regional anesthesia, making it an attractive alternative to hysteroscopy performed in the operating room setting. Further studies comparing "Rail Biopsy" to other EB techniques are needed. CONCLUSION: We showed a novel approach for hysteroscopic EB that may be particularly useful in patients with hypo/atrophic endometrium, easy to learn and with low costs. VIDEO ABSTRACT.

Humoral Immune Response to SARS-CoV-2 Spike Protein Receptor-Binding Motif Linear Epitopes.

The worldwide spread of SARS-CoV-2 has led to a significant economic and social burden on a global scale. Even though the pandemic has concluded, apprehension remains regarding the emergence of highly transmissible variants capable of evading immunity induced by either vaccination or prior infection. The success of viral penetration is due to the specific amino acid residues of the receptor-binding motif (RBM) involved in viral attachment. This region interacts with the cellular receptor ACE2, triggering a neutralizing antibody (nAb) response. In this study, we evaluated serum immunogenicity from individuals who received either a single dose or a combination of different vaccines against the original SARS-CoV-2 strain and a mutated linear RBM. Despite a modest antibody response to wild-type SARS-CoV-2 RBM, the Omicron variants exhibit four mutations in the RBM (S477N, T478K, E484A, and F486V) that result in even lower antibody titers. The primary immune responses observed were directed toward IgA and IgG. While nAbs typically target the RBD, our investigation has unveiled reduced seroreactivity within the RBD's crucial subregion, the RBM. This deficiency may have implications for the generation of protective nAbs. An evaluation of S1WT and S2WT RBM peptides binding to nAbs using microscale thermophoresis revealed a higher affinity (35 nM) for the S2WT sequence (GSTPCNGVEGFNCYF), which includes the FNCY patch. Our findings suggest that the linear RBM of SARS-CoV-2 is not an immunodominant region in vaccinated individuals. Comprehending the intricate dynamics of the humoral response, its interplay with viral evolution, and host genetics is crucial for formulating effective vaccination strategies, targeting not only SARS-CoV-2 but also anticipating potential future coronaviruses.

Mapping IgA Epitope and Cross-Reactivity between Severe Acute Respiratory Syndrome-Associated Coronavirus 2 and DENV.

BACKGROUND: The newly introduced COVID-19 vaccines have reduced disease severity and hospitalizations. However, they do not significantly prevent infection or transmission. In the same context, measuring IgM and IgG antibody levels is important, but it does not provide information about the status of the mucosal immune response. This article describes a comprehensive mapping of IgA epitopes of the S protein, its cross-reactivity, and the development of an ELISA-peptide assay. METHODS: IgA epitope mapping was conducted using SPOT synthesis and sera from RT-qPCR COVID-19-positive patients. Specific and cross-reacting epitopes were identified, and an evolutionary analysis from the early Wuhan strain to the Omicron variant was performed using bioinformatics tools and a microarray of peptides. The selected epitopes were chemically synthesized and evaluated using ELISA-IgA. RESULTS: A total of 40 IgA epitopes were identified with 23 in S1 and 17 in the S2 subunit. Among these, at least 23 epitopes showed cross-reactivity with DENV and other organisms and 24 showed cross-reactivity with other associated coronaviruses. Three MAP4 polypeptides were validated by ELISA, demonstrating a sensitivity of 90-99.96% and a specificity of 100%. Among the six IgA-RBD epitopes, only the SC/18 epitope of the Omicron variants (BA.2 and BA.2.12.1) presented a single IgA epitope. CONCLUSIONS: This research unveiled the IgA epitome of the S protein and identified many epitopes that exhibit cross-reactivity with DENV and other coronaviruses. The S protein of variants from Wuhan to Omicron retains many conserved IgA epitopes except for one epitope (#SCov/18). The cross-reactivity with DENV suggests limitations in using the whole S protein or the S1/S2/RBD segment for IgA serological diagnostic tests for COVID-19. The expression of these identified specific epitopes as diagnostic biomarkers could facilitate monitoring mucosal immunity to COVID-19, potentially leading to more accurate diagnoses and alternative mucosal vaccines.

Mapping the vulnerability of irrigation sand traps in a tropical volcanic basin, Indonesia.

Sand traps in irrigation networks are typically used in mitigating canal sedimentation. In irrigation networks located in basins of high sediment yield due to the presence of volcanoes, it is essential to assess the vulnerability of sand traps. Using sediment yield at irrigation scheme inlets, sand trap vulnerability can be evaluated. This study aims to understand the vulnerability of irrigation sand traps throughout the Progo-Opak-Serang (POS) Volcanic River Basin, Indonesia, via mapping the sediment yield distributions in the basin. We employed the Revised Universal Soil Loss Equation to estimate soil loss, where the results show that the average soil loss in the POS River Basin is 179.69 tons/ha/year that falls under the category of moderate erosion potential, while the average sediment yield for the whole basin is 51.04 tons/ha/year. Parts of the basin with high yields of more than 180 tons/ha/year were mostly found along the volcanic mountains such as Sindoro, Sumbing, Merapi, Merbabu, and Telomoyo, and the Menoreh Hills. The model demonstrated relatively high performance with R2, NSE, RMSE, and MAE of 0.89, 0.82, 0.14, and 0.11, respectively. Within the POS Basin, Badran, Kalibawang, and Blawong are the three most vulnerable irrigation sand traps, with sediment yield values of 252.83, 178.92, and 63.49 tons/ha/year, respectively; they are all located in sub-watershed outlets. The vulnerability assessment conducted in this study can be used for the decision support system to prioritize irrigation sand traps towards a more effective irrigation system development.

An Update on the Therapeutic Potential of Antimicrobial Peptides against Acinetobacter baumannii Infections.

The rise in antibiotic-resistant strains of clinically important pathogens is a major threat to global health. The World Health Organization (WHO) has recognized the urgent need to develop alternative treatments to address the growing list of priority pathogens. Antimicrobial peptides (AMPs) rank among the suggested options with proven activity and high potential to be developed into effective drugs. Many AMPs are naturally produced by living organisms protecting the host against pathogens as a part of their innate immunity. Mechanisms associated with AMP actions include cell membrane disruption, cell wall weakening, protein synthesis inhibition, and interference in nucleic acid dynamics, inducing apoptosis and necrosis. Acinetobacter baumannii is a critical pathogen, as severe clinical implications have developed from isolates resistant to current antibiotic treatments and conventional control procedures, such as UV light, disinfectants, and drying. Here, we review the natural AMPs representing primary candidates for new anti-A. baumannii drugs in post-antibiotic-era and present computational tools to develop the next generation of AMPs with greater microbicidal activity and reduced toxicity.

High-Throughput IgG Epitope Mapping of Tetanus Neurotoxin: Implications for Immunotherapy and Vaccine Design.

Tetanus is an acute, fatal disease caused by exotoxins released from Clostridium tetani during infections. A protective humoral immune response can be induced by vaccinations with pediatric and booster combinatorial vaccines that contain inactivated tetanus neurotoxin (TeNT) as a major antigen. Although some epitopes in TeNT have been described using various approaches, a comprehensive list of its antigenic determinants that are involved with immunity has not been elucidated. To this end, a high-resolution analysis of the linear B-cell epitopes in TeNT was performed using antibodies generated in vaccinated children. Two hundred sixty-four peptides that cover the entire coding sequence of the TeNT protein were prepared in situ on a cellulose membrane through SPOT synthesis and probed with sera from children vaccinated (ChVS) with a triple DTP-vaccine to map continuous B-cell epitopes, which were further characterized and validated using immunoassays. Forty-four IgG epitopes were identified. Four (TT-215-218) were chemically synthesized as multiple antigen peptides (MAPs) and used in peptide ELISAs to screen post-pandemic DTP vaccinations. The assay displayed a high performance with high sensitivity (99.99%) and specificity (100%). The complete map of linear IgG epitopes induced by vaccination with inactivated TeNT highlights three key epitopes involved in the efficacy of the vaccine. Antibodies against epitope TT-8/G can block enzymatic activity, and those against epitopes TT-41/G and TT-43/G can interfere with TeNT binding to neuronal cell receptors. We further show that four of the epitopes identified can be employed in peptide ELISAs to assess vaccine coverage. Overall, the data suggest a set of select epitopes to engineer new, directed vaccines.

The mechanisms of selective attention in phenomenal consciousness.

In three experiments we investigated the effects of selective attention in iconic memory and fragile-visual short-term memory (VSTM), which have been related to phenomenal consciousness. We used a novel retro-cue paradigm with different delays (early vs late) and object priorities (high vs equal vs low), to investigate (a) attentional costs and benefits and the role of (b) bottom-up factors and (c) fragile-VSTM in feature-based attentional selection. Experiment 1 showed that attentional costs modulate visual maintenance at longer delays, while Experiment 2 showed that by reducing the time exposure of the memory array from 250 ms to 100 ms, as a bottom-up factor, participants were not able to select the objects based on their priorities. Finally, Experiment 3 showed that a pattern mask presented before the transfer in visual working memory, attenuates the overall performance while preserving the priority effect. The implications for phenomenal consciousness before conscious access are discussed.

Effectiveness Evaluation of a UV-C-Photoinactivator against Selected ESKAPE-E Pathogens.

Healthcare-associated infections (HAI) worldwide includes infections by ESKAPE-E pathogens. Environmental surfaces and fomites are important components in HAI transmission dynamics, and shoe soles are vectors of HAI. Ultraviolet (UV) disinfection is an effective method to inactivate pathogenic microorganisms. In this study, we investigated whether the SANITECH UV-C shoe sole decontaminator equipment that provides germicidal UV-C radiation could effectively reduce this risk of different pathogens. Six standard strains and four clinical MDR strains in liquid and solid medium were exposed to a UV-C System at specific concentrations at other times. Bacterial inactivation (growth and cultivability) was investigated using colony counts and resazurin as metabolic indicators. SEM was performed to assess the membrane damage. Statistically significant reduction in cell viability for all ATCCs strains occurred after 10 s of exposure to the UV-C system, except for S. enterica, which only occurred at 20 s. The cell viability of P. aeruginosa (90.9%), E. faecalis and A. baumannii (85.3%), S. enterica (82.9%), E. coli (79.2%) and S. aureus (71.9%) was reduced considerably at 20 s. In colony count, after 12 s of UV-C exposure, all ATCC strains showed a 100% reduction in CFU counts, except for A. baumannii, which reduced by 97.7%. A substantial reduction of colonies above 3 log10 was observed at 12 and 20 s in all bacterial strains tested, except for A. baumannii ATCC 19606 (12 s). The exposure of ATCCs bacterial strains to the UV-C system for only 2 s was able to reduce 100% in the colony forming units (CFU) count in all ATCCs strains, S. aureus, P. aeruginosa, E. coli, A. baumannii, E. faecalis, except the S. enterica strain which had a statistically significant reduction of 99.7%. In ATCC strains, there was a substantial decrease in colonies after 4 s (sec) of exposure to the UV-C system, with a reduction ranging from 3.78-4.15 log10 CFU/mL. This reduction was observed in MDR/ESKAPE-E strains within 10 s, showing that UV-C could eliminate above 3.84 log10 CFU/mL. SEM showed a reduction of pili-like appendages after UV-C treatment in all strains except for E. coli (ATCC 25922). The Sanitech UV-C shoe sole decontaminator equipment from Astech Serv. and Fabrication Ltd. (Petrópolis, Brazil), effectively killed in vitro a series of ATCCs and MDR/ESKAPE-E bacteria of sanitary interest, commonly found in the hospital environment.

Evaluating aquifer stress and resilience with GRACE information at different spatial scales in Cambodia.

Groundwater exploitation for different sectors in Cambodia is expanding. Groundwater levels have already begun to decline in some parts of the country. Monitoring and assessing groundwater storage (GWS) change, aquifer stress and aquifer resilience will support the proper planning and management of the country's groundwater resources; however, information regarding groundwater in Cambodia is currently scarce. Thus, GWS change in Cambodia over the 15 years from April 2002 to March 2017 was assessed using remote-sensing-based Gravity Recovery and Climate Experiment (GRACE) and Global Land Data Assimilation System (GLDAS) datasets, with a comprehensive validation of the GRACE-derived groundwater storage anomaly (GWSA) with respect to in-situ field-based observations. The current study also investigated the impact of surface water storage (SWS) change in Tonle Sap Lake, South-East Asia's largest freshwater lake, on deriving the GWS change in Cambodia. The groundwater aquifer stresses (GAS), and aquifer resilience (AR) were also evaluated. The validation results were promising, with the correlation coefficient between satellite-based estimations and ground-based measurements ranging from 0.82 to 0.88 over four subbasins. The overall decreasing rate of GWS was found to be -0.63 mm/month, with two basins having the highest declining rate of more than 1.4 mm/month. Meanwhile, the aquifer experiencing stress during the dry season had a very low ability to quickly recover from these stresses. These findings emphasise that appropriate management is urgently needed to ensure the sustainability of the groundwater resource system in this country. Supplementary Information: The online version contains supplementary material available at 10.1007/s10040-022-02570-w.

L'exploitation des eaux souterraines s'étend au Cambodge dans différents secteurs. Les niveaux piézométriques ont déjà commencé à baisser sur quelques secteurs du pays. Surveiller et évaluer les changements de stock d'eau souterraine (GWS), la pression sur les aquifères et leur résilience supportera une planification et gestion correctes des ressources en eau dans le pays; toutefois, les informations en relation avec les eaux souterraines sont peu nombreuses aujourd'hui au Cambodge. Ainsi, les changements de GWS au Cambodge sur les 15 dernières années, d'avril 2002 à mars 2017 ont été évalués à l'aide des méthodes de télédétection basées sur le Gravity Recovery and Climate Experiment (GRACE) et les jeux de données du Global Land Data Assimilation System (GLDAS) avec une validation complète des anomalies de stockage d'eau souterraine (GWSA) de GRACE par l'utilisation d'observations de terrain. L'étude a également permis d'étudier l'impact des changements de stocks d'eau de surface (SWS) dans le lac Tonle Sap, le plus grand lac d'eau douce du sud-est asiatique, par dérivation des changements de GWS au Cambodge. Les pressions sur les eaux souterraines (GAS) et la résilience des aquifère (AR) ont également été évaluées. La validation des résultats est prometteuse, avec un coefficient de corrélation entre les estimations basées sur les données satellitaires et les mesures de terrain allant de 0.82 à 0.88 sur quatre sous-bassins. Un taux de baisse globale du GWS de ­0.63 mm/mois a été estimé, avec deux sous-bassins ayant des baisses plus fortes de plus de 1.4 mm/mois. Sur la même période, les aquifère qui subissent un stress durant les basses eaux montrent une faible capacité à récupérer de ce stress. Ces résultats montrent qu'une gestion adéquate est urgemment nécessaire pour assurer la durabilité de la ressource en eau souterraine dans ce pays.

La explotación de las aguas subterráneas para diferentes sectores en Camboya está en expansión. Los niveles de las aguas subterráneas ya han empezado a descender en algunas partes del país. El seguimiento y la evaluación de los cambios en el almacenamiento de las aguas subterráneas (GWS), el estrés del acuífero y la resiliencia del acuífero apoyarán la planificación y la gestión adecuadas de los recursos de aguas subterráneas del país; sin embargo, la información relativa a las aguas subterráneas en Camboya es actualmente escasa. Por lo tanto, se evaluó el cambio de GWS en Camboya durante los últimos 15 años, desde abril de 2002 hasta marzo de 2017, utilizando conjuntos de datos del Gravity Recovery and Climate Experiment (GRACE) y del Global Land Data Assimilation System (GLDAS) basados en la teledetección, con una validación exhaustiva de la anomalía de almacenamiento de aguas subterráneas (GWSA) derivada de GRACE con respecto a las observaciones in situ sobre el terreno. El presente estudio también investigó el impacto del cambio en el almacenamiento de agua superficial (SWS) en el lago Tonle Sap, el mayor lago de agua dulce del sudeste asiático, en la derivación del cambio del GWS en Camboya. También se evaluaron las tensiones de los acuíferos subterráneos (GAS) y la resistencia de los acuíferos (AR). Los resultados de la validación fueron promisorios, ya que el coeficiente de correlación entre las estimaciones basadas en satélites y las mediciones terrestres osciló entre 0.82 y 0.88 en cuatro subcuencas. La tasa global de disminución del GWS fue de ­0.63 mm/mes, con dos cuencas con la tasa de disminución más alta, de más de 1.4 mm/mes. Mientras tanto, el acuífero que experimentaba estrés durante la estación seca tenía una capacidad muy baja para recuperarse rápidamente de estas tensiones. Estos resultados ponen de relieve que se necesita urgentemente una gestión adecuada para garantizar la sostenibilidad del sistema de recursos hídricos subterráneos en este país.

A exploração de águas subterrâneas para diferentes setores no Camboja está se expandindo. Os níveis das águas subterrâneas já começaram a diminuir em algumas partes do país. O monitoramento e avaliação das mudanças no armazenamento de águas subterrâneas (AASub), estresse e resiliência do aquífero apoiarão o planejamento e a gestão adequados dos recursos hídricos subterrâneos do país; no entanto, as informações sobre as águas subterrâneas no Camboja são atualmente escassas. Assim, a mudança de AASub no Camboja nos últimos 15 anos, de abril de 2002 a março de 2017, foi avaliada usando conjuntos de dados Gravity Recovery and Climate Experiment (GRACE) e Global Land Data Assimilation System (GLDAS) baseados em sensoriamento remoto, com uma validação abrangente da anomalia de armazenamento de água subterrânea derivada do GRACE (AAASub) em relação a observações baseadas em campo. O estudo atual também investigou o impacto da mudança de armazenamento de água de superfície (AASup) no Lago Tonle Sap, o maior lago de água doce do Sudeste Asiático, na derivação da mudança de AASub no Camboja. Os estresses das águas subterrâneas no aquífero (SASub) e a resiliência do aquífero (RA) também foram avaliados. Os resultados da validação foram promissores, com o coeficiente de correlação entre as estimativas baseadas em satélite e as medições terrestres variando de 0.82 a 0.88 em quatro sub-bacias. A taxa global decrescente de AAS foi de ­0.63 mm/mês, com duas bacias tendo a maior taxa de declínio de mais de 1.4 mm/mês. Enquanto isso, o aquífero submetido a estresse durante a estação seca teve uma capacidade muito baixa de se recuperar rapidamente desses estresses. Esses achados enfatizam que uma gestão adequada é urgentemente necessária para garantir a sustentabilidade do sistema de recursos hídricos subterrâneos neste país.

Inequality of Low Air Quality-Related Health Impacts among Socioeconomic Groups in the World of Work.

This research aimed to assess the perceptions of air quality and health symptoms caused by low urban air quality among vulnerable socio-economic groups in the world of work in Bangkok, Thailand through a questionnaire survey of 400 workers of both formal and informal sectors in the five districts with different socio-economic characteristics and levels of air pollution. The findings showed symmetry between air quality-monitoring data and health symptoms of different socio-economic groups but asymmetry between air quality-monitoring data and people's perceptions of air quality in their areas. It also showed inequalities of low air quality-related health impacts on socio-economic groups in the world of work. People working near the streets, highways, and industrial zones tended to have more health symptoms related to low air quality, and informal sector workers faced more health risks than formal sector workers. The study appeals for effective air pollution communication to enhance the public and informal sector worker population's literacy of air pollution, the sources of air pollution and its critical health impacts, and the available and sufficient primary care organizations and community health care centers to address work-related health needs to reach the informal sector worker population.

Got milk? Maternal immune activation during the mid-lactational period affects nutritional milk quality and adolescent offspring sensory processing in male and female rats.

Previous studies have underscored the importance of breastfeeding and parental care on offspring development and behavior. However, their contribution as dynamic variables in animal models of early life stress are often overlooked. In the present study, we investigated how lipopolysaccharide (LPS)-induced maternal immune activation (MIA) on postnatal day (P)10 affects maternal care, milk, and offspring development. MIA was associated with elevated milk corticosterone concentrations on P10, which recovered by P11. In contrast, both milk triglyceride and percent creamatocrit values demonstrated a prolonged decrease following inflammatory challenge. Adolescent MIA offspring were heavier, which is often suggestive of poor early life nutrition. While MIA did not decrease maternal care quality, there was a significant compensatory increase in maternal licking and grooming the day following inflammatory challenge. However, this did not protect against disrupted neonatal huddling or later-life alterations in sensorimotor gating, conditioned fear, mechanical allodynia, or reductions in hippocampal parvalbumin expression in MIA offspring. MIA-associated changes in brain and behavior were likely driven by differences in milk nutritional values and not by direct exposure to LPS or inflammatory molecules as neither LPS binding protein nor interleukin-6 milk levels differed between groups. These findings reflected comparable microbiome and transcriptomic patterns at the genome-wide level. Animal models of early life stress can impact both parents and their offspring. One mechanism that can mediate the effects of such stressors is changes to maternal lactation quality which our data show can confer multifaceted and compounding effects on offspring physiology and behavior.

Milking It for All It's Worth: The Effects of Environmental Enrichment on Maternal Nurturance, Lactation Quality, and Offspring Social Behavior.

Breastfeeding confers robust benefits to offspring development in terms of growth, immunity, and neurophysiology. Similarly, improving environmental complexity, i.e., environmental enrichment (EE), contributes developmental advantages to both humans and laboratory animal models. However, the impact of environmental context on maternal care and milk quality has not been thoroughly evaluated, nor are the biological underpinnings of EE on offspring development understood. Here, Sprague Dawley rats were housed and bred in either EE or standard-housed (SD) conditions. EE dams gave birth to a larger number of pups, and litters were standardized and cross-fostered across groups on postnatal day (P)1. Maternal milk samples were then collected on P1 (transitional milk phase) and P10 (mature milk phase) for analysis. While EE dams spent less time nursing, postnatal enrichment exposure was associated with heavier offspring bodyweights. Milk from EE mothers had increased triglyceride levels, a greater microbiome diversity, and a significantly higher abundance of bacterial families related to bodyweight and energy metabolism. These differences reflected comparable transcriptomic changes at the genome-wide level. In addition to changes in lactational quality, we observed elevated levels of cannabinoid receptor 1 in the hypothalamus of EE dams, and sex-dependent and time-dependent effects of EE on offspring social behavior. Together, these results underscore the multidimensional impact of the combined neonatal and maternal environments on offspring development and maternal health. Moreover, they highlight potential deficiencies in the use of "gold standard" laboratory housing in the attempt to design translationally relevant animal models in biomedical research.

An allostatic epigenetic memory on chromatin footprints after double-hit acute stress.

Stress induces allostatic responses, whose limits depend on genetic background and the nature of the challenges. Allostatic load reflects the cumulation of these reponses over the course of life. Acute stress is usually associated with adaptive responses, although, depending on the intensity of the stress and individual differences , some may experience maladaptive coping that persists through life and may influence subsequent responses to stressful events, as is the case of post-traumatic stress disorder. We investigated the behavioral traits and epigenetic signatures in a double-hit mouse model of acute stress in which heterotypic stressors (acute swim stress and acute restraint stress) were applied within a 7-day interval period. The ventral hippocampus was isolated to study the footprints of chromatin accessibility driven by exposure to double-hit stress. Using ATAC sequencing to determine regions of open chromatin, we showed that depending on the number of acute stressors, several gene sets related to development, immune function, cell starvation, translation, the cytoskeleton, and DNA modification were reprogrammed in both males and females. Chromatin accessibility for transcription factor binding sites showed that stress altered the accessibility for androgen, glucocorticoid, and mineralocorticoid receptor binding sites (AREs/GREs) at the genome-wide level, with double-hit stressed mice displaying a profile unique from either single hit of acute stress. The investigation of AREs/GREs adjacent to gene coding regions revealed several stress-related genes, including Fkbp5, Zbtb16, and Ddc, whose chromatin accessibility was affected by prior exposure to stress. These data demonstrate that acute stress is not truly acute because it induces allostatic signatures that persist in the epigenome and may manifest when a second challenge hits later in life.

Recent Advances in the Immunologic Method Applied to Tick-Borne Diseases in Brazil.

Zoonotic-origin infectious diseases are one of the major concerns of human and veterinary health systems. Ticks, as vectors of several zoonotic diseases, are ranked second only to mosquitoes as vectors. Many ticks' transmitted infections are still endemic in the Americas, Europe, and Africa and represent approximately 17% of their infectious diseases population. Although our scientific capacity to identify and diagnose diseases is increasing, it remains a challenge in the case of tick-borne conditions. For example, in 2017, 160 cases of the Brazilian Spotted Fever (BSF, a tick-borne illness) were confirmed, alarming the notifiable diseases information system. Conversely, Brazilian borreliosis and ehrlichiosis do not require notification. Still, an increasing number of cases in humans and dogs have been reported in southeast and northeastern Brazil. Immunological methods applied to human and dog tick-borne diseases (TBD) show low sensitivity and specificity, cross-reactions, and false IgM positivity. Thus, the diagnosis and management of TBD are hampered by the personal tools and indirect markers used. Therefore, specific and rapid methods urgently need to be developed to diagnose the various types of tick-borne bacterial diseases. This review presents a brief historical perspective on the evolution of serological assays and recent advances in diagnostic tests for TBD (ehrlichiosis, BSF, and borreliosis) in humans and dogs, mainly applied in Brazil. Additionally, this review covers the emerging technologies available in diagnosing TBD, including biosensors, and discusses their potential for future use as gold standards in diagnosing these diseases.

Potent Activity of a High Concentration of Chemical Ozone against Antibiotic-Resistant Bacteria.

BACKGROUND: Health care-associated infections (HAIs) are a significant public health problem worldwide, favoring multidrug-resistant (MDR) microorganisms. The SARS-CoV-2 infection was negatively associated with the increase in antimicrobial resistance, and the ESKAPE group had the most significant impact on HAIs. The study evaluated the bactericidal effect of a high concentration of O3 gas on some reference and ESKAPE bacteria. MATERIAL AND METHODS: Four standard strains and four clinical or environmental MDR strains were exposed to elevated ozone doses at different concentrations and times. Bacterial inactivation (growth and cultivability) was investigated using colony counts and resazurin as metabolic indicators. Scanning electron microscopy (SEM) was performed. RESULTS: The culture exposure to a high level of O3 inhibited the growth of all bacterial strains tested with a statistically significant reduction in colony count compared to the control group. The cell viability of S. aureus (MRSA) (99.6%) and P. aeruginosa (XDR) (29.2%) was reduced considerably, and SEM showed damage to bacteria after O3 treatment Conclusion: The impact of HAIs can be easily dampened by the widespread use of ozone in ICUs. This product usually degrades into molecular oxygen and has a low toxicity compared to other sanitization products. However, high doses of ozone were able to interfere with the growth of all strains studied, evidencing that ozone-based decontamination approaches may represent the future of hospital cleaning methods.

Tumor-associated macrophages (TAMs) modulate response to HER2-targeted agents in a humanized mouse model of breast cancer

PurposeTumor-associated macrophages (TAM) may participate to antitumor activity of anti-HER2-targeted therapies (Pertuzumab, Trastuzumab) in breast cancers harbouring HER-2 overexpression through antibody-dependent phagocytosis. Additive antitumor effect of concurrent cytotoxic chemotherapies, including Paclitaxel, may be counterbalanced by alteration in TAM infiltrate. The aim of this study is to evaluate the role of TAM in tumor response to anti-HER2-targeted therapies and chemotherapy in an experimental model of HER2-amplified breast cancer.MethodsA xenograft mouse model was built by subcutaneous injection of the SKBR-3 human HER2-amplified breast cancer cell line in Hu-CD34+ mice. Animals were randomized to receive weekly administration of Cremophor (control), Trastuzumab+Pertuzumab (TP), and Paclitaxel+Trastuzumab+Pertuzumab (PTP) with or without macrophage depletion with clodronate (C). At week 4, mice were euthanised and tumors were harvested for immunohistochemical analysis of TAM infiltration (RBP-J CD163 and CD68 for M1, M2, and overall TAM, respectively).ResultsTumor size was significantly lower in mice treated with TP, PTP, and PTP+C as compared to control, while no meaningful difference was observed in the TP+C arm. Analysis of TAM infiltrate showed significantly lower CD68 and CD163 expression in PTP, TP+C, and PTP+C as compared to TP and control arm. RBP-J expression was significantly decreased in mice treated with clodronate depletion.ConclusionsActivity of TP is modulated by TAM infiltrate, that is inhibited by concurrent administration of Paclitaxel. To enhance the effect of anti-HER2-targeted therapies and minimize chemotherapy-related side effects, modulation of TAM should be considered in novel therapeutic combinations.

Rapid Detection of Anti-SARS-CoV-2 Antibodies with a Screen-Printed Electrode Modified with a Spike Glycoprotein Epitope.

BACKGROUND: The coronavirus disease of 2019 (COVID-19) is caused by an infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It was recognized in late 2019 and has since spread worldwide, leading to a pandemic with unprecedented health and financial consequences. There remains an enormous demand for new diagnostic methods that can deliver fast, low-cost, and easy-to-use confirmation of a SARS-CoV-2 infection. We have developed an affordable electrochemical biosensor for the rapid detection of serological immunoglobulin G (IgG) antibody in sera against the spike protein. MATERIALS AND METHODS: A previously identified linear B-cell epitope (EP) specific to the SARS-CoV-2 spike glycoprotein and recognized by IgG in patient sera was selected for the target molecule. After synthesis, the EP was immobilized onto the surface of the working electrode of a commercially available screen-printed electrode (SPE). The capture of SARS-CoV-2-specific IgGs allowed the formation of an immunocomplex that was measured by square-wave voltammetry from its generation of hydroquinone (HQ). RESULTS: An evaluation of the performance of the EP-based biosensor presented a selectivity and specificity for COVID-19 of 93% and 100%, respectively. No cross-reaction was observed to antibodies against other diseases that included Chagas disease, Chikungunya, Leishmaniosis, and Dengue. Differentiation of infected and non-infected individuals was possible even at a high dilution factor that decreased the required sample volumes to a few microliters. CONCLUSION: The final device proved suitable for diagnosing COVID-19 by assaying actual serum samples, and the results displayed good agreement with the molecular biology diagnoses. The flexibility to conjugate other EPs to SPEs suggests that this technology could be rapidly adapted to diagnose new variants of SARS-CoV-2 or other pathogens.

New Insights into Hemopexin-Binding to Hemin and Hemoglobin.

Hemopexin (Hx) is a plasma glycoprotein that scavenges heme (Fe(III) protoporphyrin IX). Hx has important implications in hemolytic disorders and hemorrhagic conditions because releasing hemoglobin increases the labile heme, which is potentially toxic, thus producing oxidative stress. Therefore, Hx has been considered for therapeutic use and diagnostics. In this work, we analyzed and mapped the interaction sequences of Hx with hemin and hemoglobin. The spot-synthesis technique was used to map human hemopexin (P02790) binding to hemin and human hemoglobin. A library of 15 amino acid peptides with a 10-amino acid overlap was designed to represent the entire coding region (aa 1-462) of hemopexin and synthesized onto cellulose membranes. An in silico approach was taken to analyze the amino acid frequency in the identified interaction regions, and molecular docking was applied to assess the protein-protein interaction. Seven linear peptide sequences in Hx were identified to bind hemin (H1-H7), and five were described for Hb (Hb1-Hb5) interaction, with just two sequences shared between hemin and Hb. The amino acid composition of the identified sequences demonstrated that histidine residues are relevant for heme binding. H105, H293, H373, H400, H429, and H462 were distributed in the H1-H7 peptide sequences, but other residues may also play an important role. Molecular docking analysis demonstrated Hx's association with the ß-chain of Hb, with several hotspot amino acids that coordinated the interaction. This study provides new insights into Hx-hemin binding motifs and protein-protein interactions with Hb. The identified binding sequences and specific peptides can be used for therapeutic purposes and diagnostics as hemopexin is under investigation to treat different diseases and there is an urgent need for diagnostics using labile heme when monitoring hemolysis.