Automating Wood Species Detection and Classification in Microscopic Images of Fibrous Materials with Deep Learning.

We have developed a methodology for the systematic generation of a large image dataset of macerated wood references, which we used to generate image data for nine hardwood genera. This is the basis for a substantial approach to automate, for the first time, the identification of hardwood species in microscopic images of fibrous materials by deep learning. Our methodology includes a flexible pipeline for easy annotation of vessel elements. We compare the performance of different neural network architectures and hyperparameters. Our proposed method performs similarly well to human experts. In the future, this will improve controls on global wood fiber product flows to protect forests.

Comprehensive Analysis of Physicochemical Properties and Volatile Compounds in Different Strawberry Wines under Various Pre-Treatments.

Pre-fermentation treatment has an important impact on the color, aroma, taste, and other characteristics of fruit wine. To discover suitable pre-treatment techniques and conditions that yield strawberry wine of excellent quality, the influences of juice fermentation, pulp maceration, thermovinification, and enzymatic hydrolysis pre-treatments on the basic chemical composition, color, antioxidant capacity, and volatile organic compounds in strawberry wines were investigated. The results showed that the color, antioxidant properties, and volatile aroma of strawberry wines fermented with juice were different from those with pulp. Strawberry wines fermented from juice after 50 °C maceration had more desirable qualities, such as less methanol content (72.43 ± 2.14 mg/L) compared with pulp-fermented wines (88.16 ± 7.52 mg/L) and enzymatic maceration wines (136.72 ± 11.5 mg/L); higher total phenolic content (21.78%) and total flavonoid content (13.02%); enhanced DPPH (17.36%) and ABTS (27.55%) free radical scavenging activities; richer essential terpenoids and fatty acid ethyl esters, such as linalool (11.28%), ethyl hexanoate (14.41%), ethyl octanoate (17.12%), ethyl decanoate (32.49%), and ethyl 9-decenoate (60.64%); pleasant floral and fruity notes compared with juice-fermented wines macerated at normal temperatures; and a lighter color. Overall, juice thermovinification at 50 °C is a potential pre-treatment technique to enhance the nutrition and aroma of strawberry wine.

Profile of Phenolic Compounds and Antioxidant Activity of Celery (Apium graveolens) Juices Obtained from Pulp after α-Amylase Treatment from Aspergillus oryzae.

The purpose of this study was to determine the content of certain phenolic compounds, antioxidant activity, pressing efficiency, extract content, and sugars in celeriac juices obtained from the pulp after α-amylase treatment from Aspergillus oryzae. The test material consisted of peeled and unpeeled celery pulp kept at a temperature of 25 °C with and without the enzyme for a period of 30 and 60 min. The juices obtained from them were analyzed for the content of selected phenolic acids and flavonoids using the UPLC-PDA-ESI-MS/MS method, for antioxidant activity measured using the ABTS˙+ and DPPH˙ method, and for the total polyphenol content using the F-C method. Additionally, the juice pressing efficiency, the extract content using the refractometer method, and the sugar content using the HPLC method were checked. Significantly higher antioxidant activity, pressing yield, and average content of caffeic acid glucoside, quinic acid, kaempferol-3,7-di-O-glucoside, and chrysoeriol-7-O-apiosylglucoside were obtained in juices from peeled celery. Maceration of the pulp with amylase resulted in a significant reduction in antioxidant activity compared to control samples. An is-total increase of 17-41% in total flavonoid content was observed in all juices tested after treatment with the enzyme for 30 and 60 min, and the phenolic acid content increased by 4-41% after treatment of the pulp with amylase for 60 min. The 60 min holding of the pulp at 25 °C, including with the enzyme, was shown to decrease the antioxidant activity and the content of quinic acid, ferulic acid, and chrysoriol-7-O-apiose-glucoside in the juices tested compared to the samples held for 30 min, while the content of other phenolic acids and flavonoids increased. In addition, after 60 min of enzymatic maceration, the pressing yield of the juices increased.

Ethnobotany of traditional cosmetics among the Oromo women in Madda Walabu District, Bale Zone, Southeastern Ethiopia.

Traditional cosmetics play a significant role in the cultural practices and personal hygiene of many ethnic groups worldwide. The Oromo, an indigenous community in Ethiopia, has a rich history of utilizing local plants for cosmetic purposes. However, the use of plants as traditional cosmetics has only been poorly investigated as more emphasis was given to traditional medicines. The study thus aimed to identify and document plant species, and parts used in traditional cosmetics and associated knowledge, and practices among Oromo women in the Madda Walabu district, Southeastern Ethiopia. A total of 150 Oromo women were interviewed to gather ethnobotanical information including the plant species used for cosmetics, their preparation methods, and their applications. Ethnobotanical indices were computed to determine the most important species used by the women. A total of 48 plant species belonging to 31 families used as sources of traditional cosmetics were recorded. Most of these plants were trees. Leaves were the most commonly used plant part in traditional cosmetics, and maceration and decoction were the most common preparation methods applied to prepare traditional cosmetics. Topical application of traditional cosmetics was the most common, while oral infusions were rarely reported type of administration. All categories had high informant consensus factor (ICF) values, ranging from 0.80 to 0.98. The highest ICF value was found for skin treatment, followed by the hair, and face mask. Overall, Commiphora habessinica, Gnidia stenophylla, Kalanchoe lanceolata, Mimusops kummel, Sesamothamnus rivae, Terminalia brownii, T. laxiflora, Acacia drepanolobium, A. mellifera, and Aloe citrina were the most frequently cited and culturally important plants by Oromo women for traditional cosmetics in the area. The study highlights the importance of local plant resources for maintaining cultural practices and personal hygiene. However, the cultural heritage associated with traditional cosmetics is facing threats from various factors. Therefore, a continuous effort to document and disseminate knowledge about traditional cosmetics practices to ensure their preservation and transmission and awakening younger generations about the importance of traditional cosmetics and their role in cultural heritage is crucial.

Inter- and intra-specific variations in phenotypic traits of Pectobacterium strains isolated from diverse eudicots and monocots in Taiwan.

Pectobacterium spp. are phytopathogenic bacteria whose phylogeny has been continuously revised throughout the years. Previous studies on Pectobacterium's phenotypic diversity often analyzed strains obtained from specific crops or adopted outdated Pectobacterium classification systems. Therefore, a current perspective on trait variations in Pectobacterium species or strains infecting more diverse plant species is limited. This study conducted phylogenetic and phenotypic analyses on strains isolated from 8 eudicot and 4 monocot families in Taiwan. Phylogenetic analysis on 78 strains identified 6 recognized species, namely P. brasiliense, P. aroidearum, P. actinidiae, P. colocasium, P. carotovorum, and P. versatile. Among these, the first two were the most predominant. Patterns suggesting varying host preferences among bacterial species were detected; most P. aroidearum strains were isolated from monocots, whereas P. brasiliense and P. actinidiae tended to exhibit preferences for eudicots. Physiological tests and Biolog analyses conducted on representative strains of each species revealed great within-species phenotypic variation. Despite these strain-level variations, a combination of indole production and phosphatase activity tests was capable of distinguishing all representative strains of P. brasiliense from those of other identified species. Inoculation assays on potato, bok choy, calla lily and onion showed inter- and intra-specific heterogeneities in the tested strains' maceration potentials. Virulence patterns across Pectobacterium species and strains differed depending on the inoculated host. Altogether the findings from this work expand the understanding of Pectobacterium's phenotypic diversity and provide implications for pathogen identification and management.

Rotational thrombectomy versus balloon maceration in dialysis access salvage.

BACKGROUND: Although end stage renal disease patients who undergo hemodialysis frequently experience dialysis-associated thrombotic complications, there remains a paucity of literature comparing the efficacy of different endovascular salvage techniques for grafts and fistulas. This study aims to compare primary patency in dialysis access following salvage mechanical thrombolysis/thrombectomy with a rotational thrombolysis device versus balloon maceration. METHODS: Using a database of patient records at a tertiary medical center, all patients who underwent rotational thrombectomy with the Cleaner XT™ Rotational Thrombectomy System for endovascular salvage of arteriovenous fistula or arteriovenous graft from August 2016 to July 2022 were included in the population. Fourteen patients who underwent balloon maceration for endovascular salvage were matched to the patients in the rotational thrombectomy group based on date of procedure. The primary patency of the fistula or graft for the two groups was measured. RESULTS: A total of 13 patients who underwent dialysis access salvage with rotational thrombectomy (10 grafts, 3 fistulas) were matched with 14 patients who underwent balloon maceration for dialysis access salvage (10 grafts, 4 fistulas). Six patients in both groups required additional graft access salvage (46% rotational thrombectomy; 43% balloon maceration). The median time to next dialysis salvage was 51 days for rotational thrombectomy and 43.5 days for balloon maceration (W + 9, critical value 1; compatible with statistically significant difference). CONCLUSION: Endovascular access salvage by rotational thrombectomy may provide longer primary patency compared to salvage by balloon maceration. However, a longer follow-up prospective study of a larger study population is necessary to clarify the safety and efficacy of rotational thrombectomy using the Cleaner XT™ device.

Sampling and Extraction of Plant Parasitic Nematodes.

The study of nematodes requires availability of nematode specimens and their population densities in plants and soil. This can be achieved using adequate sampling schemes and extraction methods. In this chapter, the most common and suitable sampling and extraction procedures and equipment are described. These include the use of Baermann's funnels, Cobb's decanting and sieving, floating methods such as the Oostenbrink method and Fenwick can, elutriators such as Seinhorst methods, centrifugation methods including that of Coolen, and mechanical and enzymatic maceration. The combination of different methods for cleaning the nematode suspensions is described, such as Cobb's sieving with Baermann's funnels or centrifugation, and for cysts combining Seinhorst's elutriator or Fenwick can with the alcohol methods. Methods for extraction of eggs and/or juveniles of cyst and egg mass forming nematodes, to be used as inoculum or to ascertain egg viability, are also described. Only little information is also noted on the use of molecular tools to identify and quantify nematode populations in soil and roots.

Effect of processing method (natural, washed, honey, fermentation, maceration) on the availability of heavy metals in specialty coffee.

The aim of this study was to determine the effect of various methods of processing, such as natural, washed, honey, anaerobic fermentation, and carbonic maceration, on the contents of heavy metals in green and roasted specialty coffees from various countries of origin (Ethiopia, Kenya, Rwanda, Burundi, Guatemala, Nicaragua, and Peru). The heavy metals aluminium (Al), nickel (Ni), chromium (Cr), cadmium (Cd), copper (Cu), and lead (Pb) were identified by a multi-element technique using inductively coupled plasma mass spectrometry. Mercury (Hg) content was determined by atomic absorption spectrometry. The processing method affected the contents of Hg, Al, Ni, Cr, Cd, and Pb in the green and roasted coffees (p < 0.001). Hg content varied in the green coffees processed by fermentation methods vs natural or washed methods (i.e. Rwandan and Guatemalan coffees). Cd content was highest in Guatemalan green coffee processed using carbonic maceration (0.062 mg/kg). Pb content differed between the Ethiopian and Rwandan roasted coffees, with the highest content in the washed method (0.252 mg/kg). The correlations between the contents of Cu and Al, Ni and Cr, and Pb and Cr were significant for both the roasted and green beans. In conclusion, the method of processing can affect the contents of heavy metals in green and roasted specialty coffees. Monitoring heavy metals when processing coffee with new methods, even though further processing such as roasting can substantially reduce their content in some cases, is therefore important.

Cold maceration extraction of wild fruit Terminalia bellirica (Gaertn.) Roxb.: exploring its bioactives for biomedical applications.

Terminalia bellirica (T. bellirica) (Gaertn.) Roxb. is a well-known traditional medicinal plants that show promising treatment because of fewer side effects in humans. In the present study, the total phenol, flavonoid, condensed and hydrolyzable tannins extracted and analyzed from cold macerated (CM) T. bellirica (Gaertn.) Roxb. fruit (TBF) and leaves (TBL) extract with the identification of bioactive compounds using GC-MS/MS technique. The highest amount of bioactive content was found in ethanolic extract than toluene. Current experimental data of TBF extract shows the maximum and significant biological activity like free radical scavenging activity against DPPH and FRAP assays with IC50 values of 51.07 ± 0.52 µg/ml and 63.14 ± 0.59 µg/ml respectively. However, IC50 cytotoxicity values of TBF extract on MCF-7 cells for 24 hrs was found to be 6.34 ± 0.72 µg/ml. Minimum inhibitory concentration (MIC) for infectious pathogens Escherichia coli and Bacillus cereus was >12.5 µg/ml and >100 µg/ml respectively, however, anti-inflammatory activity was demonstrated as an IC50 value of 509.1 ± 1.72 µg/ml. Cold macerated fruit extract revealed threatening inhibitory potential against the α-amylase and α-glucosidase enzymes, with IC50 of 50.98 ± 0.23 µg/ml and 46.70 ± 1.38 µg/ml respectively. Finally, the outcome of this study showed that T. bellirica (Gaertn.) Roxb. fruit extract could be an effective source of bioactives with efficient biomedical properties.

Improved Canker Processing and Viability Droplet Digital PCR Allow Detection of Erwinia amylovora Viable Nonculturable Cells in Apple Bark.

The bacterium Erwinia amylovora causes fire blight and continues to threaten global commercial apple and pear production. Conventional microbiology techniques cannot accurately determine the presence of live pathogen cells in fire blight cankers. Several factors may prevent E. amylovora from growing on solid culture media, including competing microbiota and the release of bacterial-growth-inhibitory compounds by plant material during sample processing. We previously developed a canker processing methodology and a chip-based viability digital PCR (v-dPCR) assay using propidium monoazide (PMA) to bypass these obstacles. However, sample analysis was still time-consuming and physically demanding. In this work, we improved the previous protocol using an automatic tissue homogenizer and transferred the chip-based v-dPCR to the BioRad QX200 droplet dPCR (ddPCR) platform. The improved sample processing method allowed the simultaneous, fast, and effortless processing of up to six samples. Moreover, the transferred v-ddPCR protocol was compatible with the same PMA treatment and showed a similar dynamic range, from 7.2 × 102 to 7.6 × 107 cells mL-1, as the previous v-dPCR. Finally, the improved protocol allowed, for the first time, the detection of E. amylovora viable but nonculturable (VBNC) cells in cankers and bark tissues surrounding cankers. Our v-ddPCR assay will enable new ways to evaluate resistant pome fruit tree germplasm, further dissect the E. amylovora life cycle, and elucidate E. amylovora physiology, epidemiology, and new options for canker management.

Metabolomic analysis to unravel the composition and dynamic variations of anthocyanins in bayberry-soaked wine during the maceration process.

In this work, we employed a global untargeted metabolomics technique to explore the intricate composition of anthocyanin constituents in bayberry wine and elucidate their alteration during the maceration process. Our analysis uncovered 20 distinct forms of anthocyanins in bayberry wine, including cyanidin-type, delphinidin-type, peonidin-type, malvidin-type, and other-type. 'Dongkui' (DK) bayberry wine was characterized by a predominance of glycoside forms of cyanidin-type and delphinidin-type anthocyanins, while 'Shuijing' (SJ) bayberry wine mainly contained other-type anthocyanins. Additionally, differential anthocyanins analyses conducted across various maceration periods demonstrated the different fate of the components in the wine, with a conspicuous decline in most glycosidic form anthocyanins. Moreover, correlation analysis revealed that the red hue of bayberry wine was primarily associated with cyanidin-3-O-glucoside, cyanidin-3-O-rhamnoside, delphinidin-3-O-arabinoside, and delphinidin-3-O-galactoside. This research contributes to our understanding of the anthocyanin composition and the dynamic variations in bayberry wine, opening avenues for further exploration and optimization of production techniques in the future.

Effect of Pre-fermentative Treatments on Polysaccharide Composition of White and Rosé Musts and Wines.

This paper studied the effect of conventional pre-fermentative techniques (direct pressing "CP" and cold maceration "CM") and an innovate technique (high power ultrasounds "S"), applied to Viogner and Monastrell grapes on the polysaccharide content of the musts, white and rosé wines, and after six months of bottle aging. The results showed that the longer pre-fermentation maceration time applied with the CM technique compared to the short ultrasonic maceration was key in the extraction of polysaccharides from the grape to the must. CP treatment produced wines with the lowest content of total soluble polysaccharide families since it was the least intense pretreatment for the disruption of the grape berry cell wall polysaccharides. Ultrasonic pretreatment could be used as a new tool to increase the solubilization of polysaccharides in wines, positively affecting the wine colloidal properties. During bottle aging, there wasn't a clear effect of pretreatments on the evolution of polysaccharides.

A comparison of maceration methods for the preparation of infant skeletal remains for forensic anthropological analysis.

Very little literature currently exists prescribing which maceration method to use when preparing infant human remains, resulting in bone quality that is suitable for forensic anthropological analysis. The aim of the study was to test five maceration methods to determine which is most suitable for infant remains for forensic anthropological analysis. The sample included five neonate pig carcasses (Sus scrofa domesticus), ranging between one to three days old. Five maceration methods were tested on the pig carcasses (one pig per maceration method) to determine their effectiveness. The methods included invertebrate maceration by meal worms, chemical maceration by bleach, chemical maceration by borax solution, enzymatic maceration by laundry detergent and sodium carbonate solution, and chemical maceration by sodium hypochlorite. A scoring method was created to assess the effectiveness of each maceration method. Invertebrate maceration and chemical maceration using bleach were the least successful methods of maceration (total maceration score = 8 respectively). Chemical maceration using borax and chemical maceration using sodium hypochlorite achieved complete maceration of the skeletal remains; however, they both resulted in artifacts that are unsuitable for forensic analysis (total maceration score = 14 respectively). Enzymatic maceration using laundry detergent and sodium carbonate was the most successful method (total maceration score = 17). The detergent technique subsequently successfully macerated all five sets of infant human remains. This study has validated that the enzymatic maceration technique using laundry detergent and sodium carbonate can be used to effectively macerate the remains of infant skeletal remains for forensic anthropological analysis.

Complexity of the effects of pre-fermentation oxygenation, skin contact and use of pectolytic enzymes in white winemaking as revealed by comprehensive proteomics and volatilomics analysis.

Ion exchange chromatography and SDS-PAGE followed by identification by MALDI-TOF/TOF, and two-dimensional gas chromatography with time-of-flight mass spectrometry (GC × GC-TOF-MS) were used for comprehensive proteomics and volatilomics evaluation of the effects of pre-fermentative oxygenation, skin contact and use of pectolytic enzymes in production of Malvazija istarska white wine, respectively. Many protein species and an unprecedented number of volatiles have been identified and (semi)quantified, revealing high complexity of the observed effects. Compared to a standard control wine, oxygenation treatment modulated the protein composition and resulted with a volatilome characterized by decreased levels of several important volatiles. Skin contact treatments, especially in combination with pectolytic enzymes, significantly increased the levels of a large number of proteins, but were also deprived of particular protein species found in other wines. Wines obtained by skin contact with exogenous enzymes exhibited the most complex volatile composition with increased levels of many key monoterpenoids, alcohols and esters.

Exploring for HPLC-MS/MS Profiles and Biological Activities of Different Extracts from Allium lycaonicum Siehe ex Hayek from Turkey Flora.

The present study was designed to determine the phenolic constituents, antioxidant, and enzyme inhibition activities of aerial parts and bulbs of Allium lycaonicum (family Amaryllidaceae). Extracts were prepared by maceration and Soxhlet/infusion using hexane, methanol, and water as extraction solvents. Generally, extracts from the aerial parts showed higher total phenolic and individual components and antioxidant activity than their respective bulb extracts. Maceration with water was the best to extract total phenolic content from the aerial parts (29.00 mg gallic acid equivalents (GAE)/g), while the Soxhlet extraction with hexane (22.29 mg GAE/g) was the best for the bulb. Maceration with methanol recovered the highest total flavonoid content from both the aerial parts (41.95 mg (rutin equivalents (RE)/g) and bulb (1.83 mg RE/g). Polar extracts of aerial parts were characterized by higher abundance of kaempferol-3-glucoside (≤20,624.27 µg/mg), hyperoside (≤19,722.76 µg/g), isoquercitrin (≤17,270.70 µg/g), delphindin-3,5-diglucoside (≤14,625.21 µg/g), and rutin (≤10,901.61 µg/g) than the bulb. Aerial parts' aqueous extract, prepared by maceration, exerted the highest anti-ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical activity (64.09 mg trolox equivalents (TE)/g), Cu++ (83.03 mg TE/g) and Fe+++ (63.03 mg TE/g) reducing capacity while that prepared by infusion recorded the highest anti-DPPH (2,2-diphenyl-1-picrylhydrazyl) radical (31.70 mg TE/g) and metal chelating (27.66 mg EDTAE/g) activities. The highest total antioxidant activity (1.46 mmol TE/g) was obtained by maceration of the bulb with water. Extracts obtained by organic solvents showed remarkable enzyme inhibition properties against the tested enzymes. Soxhlet extraction of the bulb with hexane and methanol recorded the highest acetylcholinesterase inhibition (4.75 mg galanthamine equivalents (GALAE)/g) and tyrosinase inhibition (139.95 mg kojic acid equivalents/g) activities, respectively. Extracts obtained by maceration of the bulb with methanol and the aerial parts with hexane exerted the highest glucosidase inhibition (3.25 mmol acarbose equivalents/g) and butyrylcholinesterase inhibition (20.99 mg GALAE/g) activities, respectively. These data indicated that A. lycaonicum is a source of bioactive molecules with potential antioxidant and enzyme inhibition properties. Nonetheless, the extracts obtained through various solvents and extraction techniques showed variations in their phytoconstituent composition and biological properties.

A comparative study on the influence of equipment design on the efficiency of dynamic maceration of Azadirachta excelsa leaves.

Dynamic maceration facilitates diffusion in solid-liquid extraction through controlling temperature and providing agitation. However, equipment design for dynamic maceration in previous investigations resulted in inadequate homogeneity of temperature and solid dispersion. A laboratory scale extractor was designed to aid the heat and mass transfer process while preventing solvent vaporization when performing dynamic maceration in a controlled environment. This study aimed to evaluate the efficiency of dynamic maceration using the laboratory scale extractor compared to a shaker incubator to extract triterpenoid saponins from Azadirachta excelsa leaves. The dynamic maceration of A. excelsa leaves was optimized using a Face-centered central composite design (FCCCD) with response surface methodology (RSM). Independent variables analyzed include ethanol-to-chloroform ratio, extraction temperature, extraction time, and sample-to-solvent ratio, while responses include yield of extract and triterpenoid saponins content (TSC). Optimum conditions were ethanol-to-chloroform ratio of 90:10, extraction temperature of 45 °C, extraction time of 60 minutes, and sample-to-solvent ratio of 1:50 g/ml. There was a significant percentage of increase in yield of extract and TSC by 41.1% and 13.3%, respectively, for the laboratory scale extractor compared to the shaker incubator. This study showed the importance of equipment design in enhancing triterpenoid saponins extraction through elevating the efficiency of the dynamic maceration process.

A maceration technique for soft plant tissue without hazardous chemicals.

Premise: Current methods for maceration of plant tissue use hazardous chemicals. The new method described here improves the safety of dissection and maceration of soft plant tissues for microscopic imaging by using the harmless enzyme pectinase. Methods and Results: Leaf material from a variety of land plants was obtained from living plants and dried herbarium specimens. Concentrations of aqueous pectinase and soaking schedules were optimized, and tissues were manually dissected while submerged in fresh solution following a soaking period. Most leaves required 2-4 h of soaking; however, delicate leaves could be macerated after 30 min while tougher leaves required 12 h to 3 days of soaking. Staining techniques can also be used with this method, and permanent or semi-permanent slides can be prepared. The epidermis, vascular tissue, and individual cells were imaged at magnifications of 10× to 400×. Only basic safety precautions were needed. Conclusions: This pectinase method is a cost-effective and safe way to obtain images of epidermal peels, separated tissues, or isolated cells from a wide range of plant taxa.

Three-dimensional analysis of the intracellular architecture by scanning electron microscopy.

The two-dimensional observation of ultrathin sections from resin-embedded specimens provides insufficient understanding of the three-dimensional (3D) morphological information of membranous organelles. The osmium maceration method, developed by Professor Tanaka's group over 40 years ago, is the only technique that allows direct observation of the 3D ultrastructure of membrane systems using scanning electron microscopy (SEM), without the need for any reconstruction process. With this method, the soluble cytoplasmic proteins are removed from the freeze-cracked surface of cells while preserving the integrity of membranous organelles, achieved by immersing tissues in a diluted osmium solution for several days. By employing the maceration method, researchers using SEM have revealed the 3D ultrastructure of organelles such as the Golgi apparatus, mitochondria, and endoplasmic reticulum in various cell types. Recently, we have developed new SEM techniques based on the maceration method to explore further possibilities for this method. These include: (1) a rapid osmium maceration method that reduces the reaction duration of the procedure, (2) a combination method that combines agarose embedding with osmium maceration to elucidate the 3D ultrastructure of organelles in free and cultured cells, and (3) a correlative immunofluorescence and SEM technique that combines cryosectioning with the osmium maceration method, enabling the correlation of the immunocytochemical localization of molecules with the 3D ultrastructure of organelles. In this paper, we review the novel osmium maceration methods described above and discuss their potential and future directions in the field of biology and biomedical research.

Polyphenolic characterisation and antiprotozoal effect of extracts obtained by maceration, ultrasound, microwave and ultrasound/microwave of Porophyllum ruderale (Jacq.) Cass.

Porophyllum ruderale (Jacq.) Cass. (Asteraceae) has antiprotozoal properties and contains extractable phenolic compounds by the maceration method (M). However, new extraction proposals such as ultrasound (U), microwaves (MW), and ultrasound/microwaves (U/MW) have emerged to optimise yields, but it is unknown if these methods modify effectiveness. Therefore, the study consisted of extracting the aerial part of P. ruderale with ethanol using the M, U, MW and U/MW methods to study its composition by RP-HPLC-ESI-MS, its total polyphenol content and its effect against Entamoeba histolytica. The study showed that U, MW and U/MW did not modify the extraction yield compared to M, but they did change the composition and the total polyphenol content. All extracts contain phloretin, caffeic acid 4-O-glucoside, todolactol A, quercetin 3-O-glucoside, quercetin 3-O-rhamnoside, luteolin and 3,7-dimethylquercetin, and affected the growth of E. histolytica. However, M and U extracts were the most effective at 5 mg/mL.

Novelty of Italian Grape Ale (IGA) beer: Influence of the addition of Gamay macerated grape must or dehydrated Aleatico grape pomace on the aromatic profile.

A new category of fruit style beer resulting from the addition of grape matrices is named Italian Grape Ale (IGA). In this paper, we report data on an experimental work to produce IGA beers, adding macerated (CO2 or N2) red Gamay grape must or Aleatico grape pomace resulting from a grape dehydration process. Our hypothesis, that these wine processes can produce volatile organic compounds (VOCs) to characterize these IGA beers which was confirmed by chemical, sensory and aromatic results. IGA beers especially the one with gas-macerated grape musts (IGA-C and IGA-N) showed higher alcohol content than ALE beer (Control) and a higher polyphenol content and antioxidant activity. As regards VOCS, IGA beers increased the concentration of some classes (i.e., alcohols, esters, norisoprenoids) and IGA-N was better characterized by specific compounds such as isobutyric acid, phenylacetate, tyrosol, ethyl hydrogen succinate. Finally, E-nose and sensory evaluation discriminated significantly all the IGA beers.