Hyperspectral analysis applied to micro-Brillouin maps of amyloid-beta plaques in Alzheimer’s disease brains

A recent investigation on the architecture and chemical composition of amyloid-β (Aβ) plaques in ex vivo histological sections of an Aβ-overexpressing transgenic mouse hippocampus has shed light on the infrared light signature of cell-activation related biomarkers of Alzheimer’s disease. A correlation was highlighted between the biomechanical properties detected by Brillouin microscopy and the molecular make-up of Aβ plaques provided by FTIR spectroscopic imaging and Raman microscopy (with correlative immunofluorescence imaging) in this animal model of the disease. In the Brillouin spectra of heterogeneous materials such as biomedical samples, peaks are likely the result of multiple contributions, more or less overlaid on a spatial and spectral scale. The ability to disentangle these contributions is very important as it may give access to discrete components that would otherwise be buried within the Brillouin peak envelope. Here, we applied an unsupervised non-negative matrix factorization method to analyse the spontaneous Brillouin microscopy maps of Aβ plaques in transgenic mouse hippocampal sections. The method has already been proven successful in decomposing chemical images and is applied here for the first time to acoustic maps acquired with a Fabry–Perot Brillouin microscope. We extracted and visualised a decrease in tissue rigidity from the core through to the periphery of the plaque, with spatially distinct components that we assigned to specific entities. This work demonstrates that it is possible to reveal the structure and mechanical properties of Aβ plaques, with details visualized by the projection of the mechanical contrast into a few relevant channels

Antibacterial Activity of Mulinum spinosum

Nasal carriers of Staphylococcus aureus are important reservoirs with risk of developing endogenous infections or transmitting infections to susceptible individuals. Methicillin-resistant S. aureus (MRSA) are associated with higher rates of treatment failure. Some strains of S. aureus produce slime which is believed to make the microorganisms more resistant to antibiotics and host defenses. The antibacterial activity of ethyl acetate : n-hexane (EtOAc : HEX) extracts of Mulinum spinosum (5 : 95% EtOAc : HEX, 50 : 50% EtOAc : HEX, 70 : 30% EtOAc : HEX and mix 20 : 80/30 : 70% EtOAc : HEX, 50 : 50/70 : 30/100 : 0% EtOAc : HEX) were assayed against 3 slime-producing S. aureus strains and 2 MRSA strains isolated from nasal carriers. S. aureus ATCC 35556 slime-producing strain and MRSA ATCC 43300 strain were used as controls. The extracts were prepared using flash chromatography. M. spinosum 5 : 95% AcOEt : HEX showed antibacterial effect against all slime-producing strains (MIC: 500 µg/mL) and the highest activity against MRSA strains (MIC: 500 to 1000 µg/mL). All M. spinosum extracts assayed were active against slime-producing S. aureus and MRSA at doses between 500 and 4000 µg/mL. Both, slime-producing S. aureus and MRSA are highly contagious and hardly eradicated by antibiotic therapies. So, there is an increasing need to find new substances with the ability to inhibit these strains

Rapid adaptation of seed germination requirements of the threatened Mediterranean species Malcolmia littorea (Brassicaceae) and implications for its reintroduction

AbstractMalcolmia littorea (Brassicaceae) is a threatened species growing in the coastal sandy dunes of the west-Mediterranean basin. In this study, the seed germination and seedling emergence requirements of this species were investigated in the only remaining native population in Italy. The highest germination percentage was achieved in darkness with scoring under safe green light at 5–10°C. Seedling emergence was highest when seeds were buried between 1 and 10mm in depth. The results suggest that germination and seedling emergence are adapted to Mediterranean coastal habitats by employing a common mechanism of light-inhibited germination and by germinating at cooler temperatures before the onset of the summer drought. Seeds were also collected from plants cultivated at a botanical garden and from plants reintroduced by sowing or by transplanting. For those populations, germination was maximal between 10–25°C, suggesting that the thermal germination behaviour may be affected by the maternal environment of seed production within one generation. It is suggested to use seeds produced in the same environment to which they will be used for the reintroduction of this species

Seasonal drought in Mediterranean soils mainly changes microbial C and N contents whereas chronic drought mainly impairs the capacity of microbes to retain P

Altres ajuts: Acord transformatiu CRUE-CSICIntensification of droughts may aggravate the generally low capacity of Mediterranean soils to store C and nutrients and induce soil C:N:P stoichiometric imbalances through its impact on soil microbial biomass and activity. Soil microbes may nonetheless have different responses to seasonal and chronic drought, but very few studies investigate long-term drought periods under field conditions. This study compares the effects of seasonal drought versus the impacts of 16 years of chronic experimental drought on microbial biomass and nutrients and assess the implications for soil nutrient availability and biogeochemical functioning in a Mediterranean forest. The chronic drought treatment reduced substantially and persistently microbial biomass C, N and particularly P, probably due to P-sparing community shifts or microbial adaptations. The smaller microbial N pool and lower mineralization activity contributed to the accumulation of C- and N-rich organic compounds in the soil and to a lower availability of mineralized forms of N during the vegetation growing season. As a result, chronic drought conditions may increase the risks of N losses from the plant-soil system in Mediterranean ecosystems. Microbial C:N ratios remained unaltered under chronic drought compared to control, likely associated with the equivalent accumulation of C- and N-rich osmolytes by microbial communities. In contrast, microbial biomass increased its C content relative to N content in response to seasonal drought, but also reduced considerably its N and P pool. Therefore, while microbial P was more sensitive to chronic water stress, microbial N and C were more closely coupled to the seasonal fluctuations of water availability

Estudio de actividad antibacteriana de extractos de Berberis ruscifolia, Baccharis sagittalis, Euphorbia dentata and Euphorbia schikendanzii, plantas nativas de Argentina y su efecto combinado

En el presente trabajo, se evaluó la actividad antibacteriana de extractos de 4 plantas nativas de la región centro-oeste argentina (Berberis ruscifolia,  Baccharis sagittalis, Euphorbia dentata and Euphorbia schikendanzii). Se seleccionaron doce extractos orgánicos para el ensayo frente a Staphylococcus  aureus, Listeria monocytogenes, Escherichia coli y Pseudomonas aeruginosa. Asimismo, se determinó el efecto combinado de los extractos acetónicos.  Todos los extractos mostraron actividad antibacteriana con valores de CIM comprendidos entre 16 y 2 mg/mL. Los extractos acetónico y clorofórmico-  metanólico de B. ruscifolia exhibieron  la mayor inhibición frente a S. aureus (CIM= 2 mg/mL). Únicamente las combinaciones de B. ruscifolia + B. sagittalis  y B. sagittalis + E. schikendanzii presentaron efecto benéfico (aditivo) para las bacterias grampositivas incluidas en este estudio

Novel Pixelwise Co-Registered Hematoxylin-Eosin and Multiphoton Microscopy Image Dataset for Human Colon Lesion Diagnosis

Colorectal cancer presents one of the most elevated incidences of cancer worldwide. Colonoscopy relies on histopathology analysis of hematoxylin-eosin (H&E) images of the removed tissue. Novel techniques such as multi-photon microscopy (MPM) show promising results for performing real-time optical biopsies. However, clinicians are not used to this imaging modality and correlation between MPM and H&E information is not clear. The objective of this paper is to describe and make publicly available an extensive dataset of fully co-registered H&E and MPM images that allows the research community to analyze the relationship between MPM and H&E histopathological images and the effect of the semantic gap that prevents clinicians from correctly diagnosing MPM images. The dataset provides a fully scanned tissue images at 10x optical resolution (0.5 µm/px) from 50 samples of lesions obtained by colonoscopies and colectomies. Diagnostics capabilities of TPF and H&E images were compared. Additionally, TPF tiles were virtually stained into H&E images by means of a deep-learning model. A panel of 5 expert pathologists evaluated the different modalities into three classes (healthy, adenoma/hyperplastic, and adenocarcinoma). Results showed that the performance of the pathologists over MPM images was 65% of the H&E performance while the virtual staining method achieved 90%. MPM imaging can provide appropriate information for diagnosing colorectal cancer without the need for H&E staining. However, the existing semantic gap among modalities needs to be corrected.This work was supported by the PICCOLO project. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under grant agreement No. 732111. The sole responsibility of this publication lies with the authors. The European Union is not responsible for any use that may be made of the information contained therein. This research has also been supported by the project ONKOTOOLS (KK2020/00069) funded by the Basque Government Industry Department under the ELKARTEK program

Viscoelasticity of amyloid plaques in transgenic mouse brain studied by Brillouin microspectroscopy and correlative Raman analysis

Amyloidopathy is one of the most prominent hallmarks of Alzheimer’s disease (AD), the leading cause of dementia worldwide, and is characterized by the accumulation of amyloid plaques in the brain parenchyma. The plaques consist of abnormal deposits mainly composed of an aggregation-prone protein fragment, β-amyloid 1-40/1-42, into the extracellular matrix. Brillouin microspectroscopy is an all-optical contactless technique that is based on the interaction between visible light and longitudinal acoustic waves or phonons, giving access to the viscoelasticity of a sample on a subcellular scale. Here, we describe the first application of micromechanical mapping based on Brillouin scattering spectroscopy to probe the stiffness of individual amyloid plaques in the hippocampal part of the brain of a β-amyloid overexpressing transgenic mouse. Correlative analysis based on Brillouin and Raman microspectroscopy showed that amyloid plaques have a complex structure with a rigid core of β-pleated sheet conformation (β-amyloid) protein surrounded by a softer ring-shaped region richer in lipids and other protein conformations. These preliminary results give a new insight into the plaque biophysics and biomechanics, and a valuable contrast mechanism for the study and diagnosis of amyloidopathy

Novel Pixelwise Co-Registered Hematoxylin-Eosin and Multiphoton Microscopy Image Dataset for Human Colon Lesion Diagnosis

Colorectal cancer presents one of the most elevated incidences of cancer worldwide. Colonoscopy relies on histopathology analysis of hematoxylin-eosin (H&E) images of the removed tissue. Novel techniques such as multi-photon microscopy (MPM) show promising results for performing real-time optical biopsies. However, clinicians are not used to this imaging modality and correlation between MPM and H&E information is not clear. The objective of this paper is to describe and make publicly available an extensive dataset of fully co-registered H&E and MPM images that allows the research community to analyze the relationship between MPM and H&E histopathological images and the effect of the semantic gap that prevents clinicians from correctly diagnosing MPM images. The dataset provides a fully scanned tissue images at 10x optical resolution (0.5 m/px) from 50 samples of lesions obtained by colonoscopies and colectomies. Diagnostics capabilities of TPF and H&E images were compared. Additionally, TPF tiles were virtually stained into H&E images by means of a deep-learning model. A panel of 5 expert pathologists evaluated the different modalities into three classes (healthy, adenoma/hyperplastic, and adenocarcinoma). Results showed that the performance of the pathologists over MPM images was 65% of the H&E performance while the virtual staining method achieved 90%. MPM imaging can provide appropriate information for diagnosing colorectal cancer without the need for H&E staining. However, the existing semantic gap among modalities needs to be corrected.This work was supported by the PICCOLO project. This project has received funding from the European Union's Horizon 2020 Research and Innovation Programme under grant agreement No. 732111. The sole re- sponsibility of this publication lies with the authors. The European Union is not responsible for any use that may be made of the information contained therei

Long-term effects of gasification biochar application on soil functions in a Mediterranean agroecosystem: Higher addition rates sequester more carbon but pose a risk to soil faunal communities

17 Pág. Departamento de Medio Ambiente y Agronomía​ (INIA)Biochar applications can have important implications for many of the soil functions upon which agroecosystems rely, particularly regarding organic carbon storage. This study evaluated the impacts of adding a highly aromatic gasification biochar at different rates (0, 12 and 50 t ha-1) to a barley crop on the provision of crucial soil functions (carbon sequestration, water content, greenhouse gas emissions, nutrient cycling, soil food web functioning, and food production). After natural ageing in the field for six years, a wide range of soil properties representative of the studied soil functions were measured and integrated into a soil quality index. Results showed that C sequestration increased with biochar rate (23 and 68% higher than in the control for the 12 and 50 t biochar ha-1 treatments, respectively). Water content was enhanced at the 50 t ha-1 treatment depending on the sampling date. Despite biochar additions neither abating nor increasing CO2 equivalent emissions (carbon dioxide plus nitrous oxide and methane), the system shifted from being a methane sink (-0.017 ± 0.01 mg CH4-C m-2 h-1 at the 12 t ha-1 treatment), to a net source (0.025 ± 0.02 mg CH4-C m-2 h-1 at the 50 t ha-1 treatment). In addition, biochar ageing provoked a loss of nitrate mitigation potential, and indeed ammonium production was stimulated at the 50 t ha-1 rate. The 50 t ha-1 treatment also adversely affected nematode and collembolan functional diversity. Lastly, biochar did not affect barley yield. The results of the soil quality index indicated that no biochar treatment provided more benefits to our agricultural soil, and, although the 50 t ha-1 treatment increased C sequestration, this was potentially offset by its harmful effects on soil faunal communities. Therefore, application of this biochar at high rates should be avoided to prevent risks to soil biological communities.We gratefully acknowledge the funding by the project FERTICHAR (AGL2015-70393-R) of the Spanish Ministry of Economy and Competitiveness. We are also grateful to Dr. van den Brink, P. J., and Dr. Šmilauer, P. for their assistance on PRC analysis interpretation and proper use of CANOCO 5, respectively.Peer reviewe

Detection of Aβ plaque-associated astrogliosis in Alzheimer’s disease brain by spectroscopic imaging and immunohistochemistry

Recent work using micro-Fourier transform infrared (μFTIR) imaging has revealed that a lipid-rich layer surrounds many plaques in post-mortem Alzheimer’s brain. However, the origin of this lipid layer is not known, nor is its role in the pathogenesis of Alzheimer’s disease (AD). Here, we studied the biochemistry of plaques in situ using a model of AD. We combined FTIR, Raman and immunofluorescence images, showing that astrocyte processes co-localise with the lipid-ring surrounding many plaques. We used μFTIR imaging to rapidly measure chemical signatures of plaques over large fields of view, and selected plaques for higher resolution analysis with Raman. Raman maps showed similar lipid-rings and dense protein cores as in FTIR images, but also revealed cell bodies. We confirmed the presence of plaques using amylo-glo staining, and measured astrocytes using immunohistochemistry, revealing astrocyte colocalisation with lipid-rings. This work is important because it correlates biochemically changes surrounding the plaque with the biological process of astrogliosis