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Pilostyles, an endoparasitic genus within the Apodanthaceae family, grows inside host stems with flowers and fruits being the only external manifestations. Previous studies of P. berteroi growing on Adesmia trijuga provided limited details of the endophyte and omitted the origin of flowers and sinker structure. This study, using classical methods of optical microscopy applied to the analysis with scanning electron microscopy and confocal laser scanning microscopy, expands the understanding of the P. berteroi/A. trijuga complex. We find that P. berteroi develops isophasically with its host, forming endophytic patches between the host's secondary phloem cells. The parasitized Adesmia stem's cambium primarily produces xylem parenchyma, with limited vessel production and halting fiber formation. The radial polarization of endophytic patches led to the formation of floral meristems. Flowers develop endogenously and emerge by the breakthrough of the host stem. Flowers are connected to the host cambium via chimeric sinkers, combining P. berteroi parenchyma and tracheoids with Adesmia vessels. Unlike previous studies that show uniformity among Pilostyles species, our analysis reveals new insights into the structural interaction between P. berteroi and A. trijuga. [ABSTRACT FROM AUTHOR]

This study aimed to develop and evaluate a niosomal formulation loaded with Plumbago zeylanica extract (PZE). The solvent evaporation method was used to prepare niosomes and optimized using the Box–Behnken design. The optimized formulation (PZE-Ns-Opt) was characterized for drug release, DPPH assay, α-amylase inhibition assay, α-glucosidase inhibition assay, and confocal laser scanning microscopy (CLSM) study. PZE-Ns-Opt showed a vesicle size of 253.6 nm, PDI of 0.108, entrapment efficiency of 62.4%, and drug release of 84.01%. The CLSM image of the rat's intestine suggested that the rhodamine red B-loaded PZE-Ns-Opts showed superior penetration compared to the control. Further, the antioxidant activity of the prepared formulation was exhibited as 89.46%±0.016 as compared to PZE (78.10%±0.005). In addition to this, α- amylase activity was inhibited by 95.11%±4.62, 85.88%±2.56, and 89.87%±3.65 by acarbose, PZE, and PZE-Ns-Opt, respectively. In comparison, the α-glucosidase activity was inhibited by 88.47%±1.04, 81.07%±0.50 and 85.78%±0.71 by acarbose, PZE, and PZE-Ns-Opt, respectively. To conclude, the development of PZE-Ns-Opt formulation and its characterization showed the establishment's authenticity. The in vitro antidiabetic, antioxidant studies expressed the protective effect from oxidative stress, CLSM of rat's intestine ability to penetrate to great extent and could be a promising candidate for the management of diabetes mellitus. [ABSTRACT FROM AUTHOR]

Aim: The osteogenic potential of new premixed calcium‐silicate‐containing bioceramic sealers (Ca‐Si sealers) was tested with porcine vascular wall‐mesenchymal stem cells (pVW‐MSCs). Methodology: Two Ca‐Si‐containing sealers: Ceraseal (MetaBiomed, Cheong‐si, South Korea) and AH Plus Bioceramic (Maruchi, Wonju‐si, South Korea), and an epoxy resin sealer (AH Plus; Dentsply, Konstanz, Germany) as a control, were prepared according to the manufacturers' indications. All samples were allowed to set for 100% of their setting time in a sterile humid cabinet at 37°C and 95% relative humidity. pVW‐MSC seeding efficiency and osteogenic differentiation were analysed as marker of gene/protein expression for up to 12 days. Mineralization assay and immunofluorescence staining were performed and evaluated over a period of 21 days. Statistical analyses were conducted using one‐way analysis of variance (p <.05). Additional samples were prepared and stored under the same conditions and inspected using an environmental scanning electron microscope equipped with an energy dispersive X‐ray spectroscopy system. Results: Significantly higher cell seeding efficiency (p <.05) was observed for both Ca‐Si sealers from day 8. pVW‐MSCs showed a significant shift towards the osteogenic lineage only when seeded in contact with Ca‐Si sealers. Gene expression of osteopontin was upregulated significantly. Collagen I and osteocalcin were clearly expressed by cells in contact with Ca‐Si sealers. Mineralization granules were observed in Alizarin red assays and confocal laser scanning microscopy analysis of both Ca‐Si sealers. No gene expression or granule mineralization were observed on the epoxy resin sealer. Conclusions: Premixed Ca‐Si sealers displayed a higher potential for osteogenic activity on pVW‐MSCs. Epoxy resin sealer was unable to induce any osteogenic activity. The properties of both Ca‐Si sealers suggest their potential as osteoinductive platforms for vascular MSCs in periapical bone. [ABSTRACT FROM AUTHOR]

In recent years, fluorescence microscopy has revitalized the study of invertebrate comparative morphology. Here we explore the usefulness of combining confocal laser scanning microscopy (CLSM) and cuticular autofluorescence to examine the taxonomically challenging marine planktonic "y-larvae" (Pancrustacea: Facetotecta). To gauge the effectiveness of CLSM with autofluorescence in producing taxonomically useful images, we applied it to seven distinct y-naupliar species or morphospecies that had previously undergone scrutiny by other techniques. The specimens were part of a museum collection of glycerin-jelly slides of exuviae of last-stage y-nauplii, a key instar for studying the taxonomy of y-larvae. For Hansenocaris demodex, the level of detail obtained from a single specimen was comparable to that previously obtained by scanning electron microscopy (SEM). For Hansenocaris aquila, revisiting the original holotype specimen resulted in a dramatic increase in our understanding of the species' morphology, including taxonomically pivotal information about its spinose labrum and a digitally rotated lateral view. CLSM analyses of the other five specimens, which represented a broad spectrum of y-naupliar morphology, efficiently generated more such information. Novel observations were made concerning putative external rudiments of both the first and second maxillae in late nauplii as well as the extreme dorso-ventral flattening of some naupliar types. CLSM observation of museum slides of naupliar exuviae using cuticular autofluorescence thus shows great promise of becoming an excellent tool for studying the morphology and taxonomy of y-larvae, and we suggest that this technique might also profitably be applied to other forms of larval exuviae. [ABSTRACT FROM AUTHOR]

Three-dimensional obturation of the root canal system is the primary goal of endodontic treatment. Endodontic sealers should fill any irregularities in the root canal system and penetrate as deeply as possible into the dentinal tubules. They should also ensure that the gutta-percha points adhere well to the dentinal walls of the root canal without leaving voids at their margins. The purpose of this study was to evaluate contemporary imaging techniques used for in vitro testing of selected specimens consisting of tooth scrapings after endodontic treatment with gutta-percha and selected endodontic sealers: AH Plus-Dentsply Sirona, Well Root ST-Vericon, Korea, and AH Bioceramics Sealer-Dentsply Sirona. The following imaging techniques were used in the evaluation: scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), micro-computed tomography (micro-CT), surgical microscopy, and light microscopy. A comparison of the imaging methods (SEM, CLSM, micro-CT, surgical microscopy, light microscopy) showed significant differences in their suitability for assessing the penetration depth of endodontic sealers into dentinal tubules in vitro. Based on the analysis of the imaging methods, it was concluded that confocal microscopy and SEM were the most objective and accurate methods for visually assessing the quality of root canal filling in vitro. However, the high cost of specimen preparation and the limited availability of specialized equipment are factors that limit their use in clinical settings. [ABSTRACT FROM AUTHOR]

Aim: The aim of the study was to evaluate and compare the depth of penetration of adhesive resin in resin-modified glass ionomer cement (RMGIC) after different surface pretreatment methods using a confocal laser scanning microscope (CLSM). Methods: Class I cavity preparation was done in 45 freshly extracted premolars and divided into three groups, according to the surface pretreatment of RMGIC which was placed in a thickness of 1 mm in the cavity. In the control group, no pretreatment was done, and other two groups were pretreated with acid etching and air abrasion (AA). The fifth-generation dentin bonding agent mixed with rhodamine B dye 0.1% was applied on RMGIC in all the samples and cured for 15 s. Samples were restored using composite resin and sectioned longitudinally. The depth of penetration of adhesive resin was evaluated using CLSM. Results: Depth of penetration of adhesive resin was highest with AA (153.70 ± 10.23), followed by acid etching (122.71 ± 12.25) and control group (77.12 ± 6.37). Conclusion: Based on the findings of this research, AA enhances the depth of penetration of adhesive resin in the RMGIC surface. Thus, AA before placement of composite resin on the RMGIC can be effective in a clinical scenario. [ABSTRACT FROM AUTHOR]

Infectious diseases, including urinary tract infections (UTIs), significantly impact global health, diminishing the quality of life and increasing clinical and economic burdens. Concerns arise due to the potential development of antimicrobial resistance (AMR) facilitated by biofilm formation. Nanotechnology, specifically silver and silver nanoparticles (1-100 nm), proves effective against bacterial strains, making them valuable in the biomedical field. Advances in microscopic imaging techniques, such as Confocal Laser Scanning Microscopy (CLSM), enhance our understanding of biofilms. This study aims to assess the effectiveness and optimal concentration of gold and silver nanoparticles in combating biofilms formed by uropathogenic Escherichia coli (UPEC) through CLSM analysis. The research comprises the following stages: rejuvenation of bacterial isolates and inoculum preparation, testing the anti-biofilm activity of gold and silver nanoparticles at concentrations of 25, 50, 75, and 100 ppm against UPEC using CLSM analysis. The findings reveal that both gold and silver nanoparticles exhibit anti-biofilm activity against UPEC at concentrations of 25, 50, 75, and 100 ppm, as confirmed by CLSM analysis. The optimal concentration for anti-biofilm activity of gold and silver nanoparticles against UPEC is identified as 100 ppm. [ABSTRACT FROM AUTHOR]

Background: The specificity in the detection of atypical and malignant melanocytic lesions in the context of skin cancer screening is very low, so that numerous unnecessary operations are performed. The extent to which complementary diagnosis using confocal laser scanning microscopy (CLSM, RCM) in skin cancer diagnosis can reduce the number of excisions is investigated. Patients and Methods: Prospective real-world study conducted in an outpatient dermatological practice. Out of 521 consecutive patients with an average age of 56 years, 173 atypical melanocytic lesions were examined using a three-step diagnostic chain: clinical examination, digital dermatoscopy, and confocal laser scanning microscopy. Results: After supplementary confocal microscopic examination, only 18 of the 173 suspicious melanocytic lesions were excised. 89.6 % of the previously planned excisions could be saved. 12 of the 18 excised lesions were dysplastic, a Spitz nevus or a melanoma in situ in the dermatohistopathological findings. Conclusion: By reducing skin surgeries in our study by nearly 90%, it was demonstrated that patients in an outpatient dermatological practice significantly benefit from the additional pain-free, imaging histological diagnostics at the cellular level using Confocal Laser Scanning Microscopy in skin cancer early detection. This diagnostic method needs to be learned and could be widely used for a substantially improved skin cancer screening. [ABSTRACT FROM AUTHOR]

This paper provides a comprehensive revision of the Palearctic plant bug genus Anapus Stål, 1858. Despite a limited number of included species, identification in this group remains challenging due to significant intraspecific polymorphism in the coloration and body proportions. To tackle this issue, confocal microscopy was used to study the male genitalia, and three-dimensional reconstructions of the aedeagus were obtained for all valid species. Our results reveal advantages of these techniques in visualizing small and complex genital structures over traditional methods of taxonomic imaging. A detailed study of the male and female genitalia resulted in finding novel characters allowing for unequivocal species recognition. The differential diagnosis for the genus and a key to all species are given. Habitus photographs, illustrations of male genitalic structures, scanning micrographs of morphological structures, host and distributional information are provided for all species. Anapus longicornis Jakovlev, 1882 is synonymized with Anapus pachymerus (Reuter, 1881) and a lectotype for the latter species is designated. Anapus americanus Knight, 1959 is transferred to the genus Myrmecophyes Fieber, 1870. [ABSTRACT FROM AUTHOR]

Elimination of microbes in the root canal system is crucial for achieving long-term success in endodontic treatment. Further efforts in study design and standardization are needed in order to improve the validity and comparability of in vitro results on endodontic disinfection procedures, in turn improving clinical outcomes. This study optimizes two models at all steps: tooth selection, pretreatment, inoculation method (by growth or centrifugation), and confocal laser scanning microscopy (CLSM)-guided imaging of LIVE/DEAD-stained specimens. Individual anatomical conditions lead to substantial differences in penetration depth. Sclerosis grading (SCG), a classification system introduced in this study, provides information about the sclerosis status of the dentine and is helpful for careful, specific, and comparable tooth selection in in vitro studies. Sonically activated EDTA for the pretreatment of roots, inoculation of Enterococcus faecalis in an overflow model, 3–4 weeks of incubation, as well as polishing of dentine slices before staining, led to advances in the visualization of bacterial penetration and irrigation depths. In contrast, NaOCl pretreatment negatively affected performance reproducibility and should be avoided in any pretreatment. Nonsclerotized teeth (SCG0) can be used for microbial semilunar-shaped inoculation by centrifugation as a "quick-and-dirty" model for initial orientation. In conclusion, CLSM-guided imaging for quantifying endodontic infection/disinfection is a very powerful method after the fine-tuning of materials and methods. [ABSTRACT FROM AUTHOR]

During blending between the binders in hot mix recycling, two physical processes occur, namely blending due to diffusion and blending due to mechanical agitation. In the present study, an experimental approach is developed to observe these processes directly on the laboratory recycled asphalt mix samples. Atomic Force Microscopy (AFM) and Confocal Laser Scanning Microscopy (CLSM) are used as the probing tools. The binder concentration profile is estimated at different locations around the reclaimed asphalt pavement material (RAPM) aggregate. Diffusion is observed to be predominant in some of these locations, while, in some other locations, mechanical mixing is observed to be predominant. Assuming that pure Fickian diffusion is happening at those diffusion-spots, the coefficient of diffusion is estimated from the concentration profile at those locations. It is envisaged that the proposed approach would provide a microscopic perspective of the extent of blending during the recycling process. [ABSTRACT FROM AUTHOR]

Gastrotrichs are highly diverse and abundant in all aquatic ecosystems; however, they are often overlooked. During a biodiversity survey in Sardinia (Italy), a new species of gastrotrich herein described was discovered. Specimens of Urodasys bifidostylis sp. nov. were found in sandy sediments from two submarine caves. Using an integrative approach of traditional light (DIC) and high-resolution (CLSM) microscopies, we herein reveal, for the first time, the fine structure and function of the reproductive organ in an Urodasys representative. This is particularly relevant considering the complex reproductive organs and strategies of this group. Results allow comparisons between the reproductive apparatus and sperm transfer modalities in Urodasys and the closely related genus Macrodasys. One similarity is that both groups transfer male gametes in packets, suggesting the production of spermatophores to be a common phenomenon in Gastrotricha. Unique to Urodasys is the ability of multiple and consecutive copulations and sperm transfers and, differently than Macrodasys, the transfer of sperms unlikely occurs simultaneously between the two hermaphroditic partners. These findings provide new insights into the reproductive strategies of Urodasys and are expected to advance future studies on the evolution of reproductive strategies and the rise of interspecific reproductive barriers in interstitial meiofauna. [ABSTRACT FROM AUTHOR]

Ex-vivo confocal laser scanning microscopy provides a rapid alternative to routine histological processing using haematoxylin and eosin-stained sections. Previous studies suggest high diagnostic accuracy in basal cell carcinoma. This study investigates the diagnostic accuracy of confocal laser scanning microscopy reporting of basal cell carcinoma in a real-life setting and compares reporting by dermatopathologists inexperienced in use of confocal laser scanning microscopy with reporting by an expert in confocal laser scanning microscopy. A total of 334 confocal laser scanning microscopy scans were diagnosed by 2 dermatopathologists inexperienced in the diagnosis of confocal laser scanning microscopy as well as an experienced examiner of confocal laser scanning microscopy scans. The inexperienced examiners achieved a sensitivity of 59.5/71.1% and specificity of 94.8/89.8%. The experienced examiner achieved a sensitivity of 78.5% and specificity of 84.8%. Detection of tumour remnants in margin controls showed insufficient values among inexperienced (30.1/33.3%) and experienced (41.7%) investigators. The results of this study, of real-life setting basal cell carcinoma reporting with confocal laser scanning microscopy, found a lower diagnostic accuracy than published data regarding artificial settings. A poor accuracy in tumour margin control is clinically relevant and could restrict the use of confocal laser scanning microscopy in clinical routine. Prior knowledge of haematoxylin and eosin trained pathologists can be partially transferred to the reporting of confocal laser scanning microscopy scans; however, specific training is recommended. [ABSTRACT FROM AUTHOR]

Aqueous gels based on two high amylose corn starch (HACS) genotypes with different amylose (AM) contents (HYLON V [HACS‐50] and HYLON VII [HACS‐70]) are prepared by gradation of the polymer concentration (6 and 9% w/w) and storage duration (1 and 14 days). The supermolecular structure of the wet gels is investigated by means of confocal laser scanning microscopy (CLSM), and the dried starch gel material examines crystallographically using X‐ray diffraction (XRD) and morphologically using scanning electron microscopy (SEM). In particular CLSM reveals that the gel structure shows changes with the storage time. XRD experiments reveal the existence of the B‐type crystalline structure as well as a remarkable portion of the V‐type, and higher crystallinity (Xc) is found after long‐term storage. Observation of the dried gel material's surface facilitates the assumption of a fractal‐based arrangement (structure of retrograded starch) within the starch gel network. The results are discussed closely with obtained data related to the molecular level of the gel matrix involved starch polymers, which are reported in previous publications. [ABSTRACT FROM AUTHOR]

Background: Digitally stained ex vivo confocal laser scanning microscopy (CLSM) scans are a possible alternative to formalin-fixed and paraffin-embedded (FFPE) and hematoxylin-eosin (H&E) stained slides. This study explores the diagnostic accuracy of digitally-stained CLSM scans in comparison to H&E-stained slides in various dermatologic diseases in a real-life setting. Methods: Samples of patients out of one selected dermatologic office were primarily scanned via CLSM; a diagnosis was made afterwards using FFPE- and H&E-stained slides by two experienced dermatopathologists. Primary outcomes were sensitivity and specificity of diagnosis in digitally stained CLSM scans in three separate diagnostic groups. Results: CLSM evaluation of epithelial tumors (n = 132) demonstrated a sensitivity of 64.3%/83.9% and a specificity of 84.2%/71.1%. Diagnosis of melanocytic tumors (n = 86) showed a sensitivity of 19.1%/85.1% and a specificity of 96.3%/66.7%. In the diagnosis of other tumors/cysts and inflammatory dermatoses (n = 42), a sensitivity of 96.4%/96.8% and a specificity of 57.1%/45.5% was reached. Conclusions: This study shows the possibilities and limitations of a broad use of CLSM. Because of a partly low diagnostic accuracy, such an application does not seem to be recommendable at present for every indication. [ABSTRACT FROM AUTHOR]

Styrene–butadiene–styrene (SBS) polymer is extensively employed for asphalt pavement construction, and its degradation significantly damages the durability of asphalt concrete. However, the effect of aging protocols on the degradation of SBS polymer in asphalt binder has not been thoroughly investigated. In this study, confocal laser scanning microscopy (CLSM) was applied to characterize the change in morphology with SBS polymer degradation. Various aging protocols were considered, including accelerated aging processes in laboratory- and field-aged samples from three highway sections with different in-service periods. Scanned images of the polymer phase in the 2D plane at different depths were processed and further reconstructed in three dimensions. Furthermore, the three-dimensional polymer morphology indices derived from the semi-quantitative analysis of the images were correlated with the rheological indices. The results show that the polymer particles change from a relatively large ellipsoidal shape to a relatively small spherical shape as aging proceeds. The increase in aging temperature appears to accelerate the degradation of the polymer at the same rheological level. The effect of the laboratory aging method on the polymer was more pronounced during the early stages of aging compared to that in the field aging process. [ABSTRACT FROM AUTHOR]

[Display omitted] • Dynamic processes of biofilm formation and cereulide production under four different conditions. • The substrate has an impact on the formation of biofilms and cereulide of Bacillus cereus. • The cereulide levels in biofilms formed in BHI medium are mostly higher than those formed in UHT milk. Bacillus cereus is a well-known foodborne pathogen that can cause human diseases, including vomiting caused by emetic toxin, cereulide, requiring 105-108 cells per gram to cause the disease. The bacterial cells may be eliminated during processing, but cereulide can survive in most processing techniques due to its resistance to high temperatures, extreme pH and proteolytic enzymes. Herein, we reported dynamic processes of biofilm formation of four different types and cereulide production within the biofilm. Confocal laser scanning microscopy (CLSM) images revealed that biofilms of the four different types reach each stage at different time points. Among the extracellular polymeric substances (EPS) components of the four biofilms formed by the emetic B. cereus F4810/72 strain, proteins account for the majority. In addition, there are significant differences (p < 0.05) in the EPS components at the same stage among biofilms of different types. The time point at which cereulide was first detected in the four types of biofilms was 24 h. In the biofilm of B. cereus formed in ultra-high-temperature (UHT) milk, the first peak of cereulide appeared at 72 h. The cereulide content of the biofilms formed in BHI was mostly higher than that of the biofilms formed in UHT milk. This study contributes to a better understanding of food safety issues in the industry caused by biofilm and cereulide toxin produced by B. cereus. [ABSTRACT FROM AUTHOR]

The green synthesis method of was used for the synthesis of silver nanoparticles using Camellia sinensis (green tea). The Camellia sinensis silver nanoparticles (CS-AgNPs) were characterized using different techniques, including UV-Vis (ultra violet-visible), SEM (scanning electron microscopy), TEM (transmission electron microscopy), and XRD (X-ray diffraction). The average size of the CS-AgNPs was 52 nm, according to TEM. The CS-AgNPs showed excellent antibacterial and antifungal activity. The MIC (minimum inhibitory concentration) against bacterial isolates varied from 31.25 to 62.5 µg/mL, whereas for fungal isolates, the MIC varied from 125 to 250 µg/mL. The presence of a zone in the well diffusion assay showed the antimicrobial nature of CS-AgNPs. Further, CLSM (confocal laser scanning microscopy) showed that CS-AgNPs possess antibiofilm activity. The interaction of CS-AgNPs with the Candidal cells was analyzed using TEM, and it was revealed that CS-AgNPs entered the cell and disrupted the cell machinery. [ABSTRACT FROM AUTHOR]

The present contribution provides the first exploration of the Loricifera fauna in New Zealand. A new species, Pliciloricus apteryx sp. nov. and a new species and genus, Patuloricus tangaroa gen. et sp. nov., are described from Bay of Plenty at North Island. The latter species is represented by a Shira larva containing a different larval stage. This is only the second loriciferan species with a Shira larva, and the first reporting of the life cycle stage following the Shira larva. This new life cycle stage is interpreted as a postlarva, and is characterized by a large globular head without external appendages, and a slender, highly muscular trunk. Both species descriptions are based on combined imaging from light microscopy and confocal laser scanning microscopy. The latter has never previously been applied to any taxonomic studies of Loricifera, but proved extremely useful resulting in the detection of structures that otherwise would have passed unnoticed. Likewise, 3D reconstructions of Z-stacks enabled a more precise interpretation of spatial arrangements of introvert and trunk appendages. [ABSTRACT FROM AUTHOR]

It is well documented that most bituminous binders contain crystallisable material. This crystallisable fraction, often referred to as paraffinic or natural wax, is associated with the bitumen's origin and has an influence on its rheological performance. In the literature, Differential Scanning Calorimetry (DSC) is the primary technique used to determine the melting and crystallisation behaviour of this waxy fraction. However, in bitumen, thermal transitions in DSC are typically very broad and can be combined with recrystallisation effects upon reheating. This work explores the potential crystallisation and melting process of a waxy and a wax-free bitumen via three different approaches: DSC, Wide-Angle X-ray Diffraction (WAXD) and Confocal Laser Scanning Microscopy (CLSM). The findings reveal that the DSC transitions of the waxy bitumen are in good agreement with the corresponding occurrence of WAXD signals and to some extent with the formation and disappearance of the surface microstructures which were followed at two cooling and heating rates. WAXD results additionally demonstrate that the crystalline material in bitumen is organised in an orthorhombic unit cell, typical for straight chain aliphatic structures. On the other hand, DSC and WAXD support the lack of crystallinity for the wax-free bitumen which could explain its featureless CLSM surface. Overall, the originality of this work resides in the disclosure of connections between crystallographic properties, thermal transitions and the surface micromorphology of bitumen. [ABSTRACT FROM AUTHOR]

The increased resistance of bacteria to antimicrobials, as well as the growing interest in innovative and sustainable alternatives to traditional food additives, are driving research towards the use of natural food preservatives. Among these, hydrolates (HYs) have gained attention as "mild" alternatives to conventional antimicrobial compounds. In this study, the response of L. monocytogenes ATCC 7644 exposed to increasing concentrations of Coridothymus capitatus HY (CHY) for 1 h at 37 °C was evaluated by means of Phenotype Microarray, modelling the kinetic data obtained by inoculating control and treated cells into GEN III microplates, after CHY removal. The results revealed differences concerning the growth dynamics in environmental conditions commonly encountered in food processing environments (different carbon sources, pH 6.0, pH 5.0, 1–8% NaCl). More specifically, for treated cells, the lag phase was extended, the growth rate was slowed down and, in most cases, the maximum concentration was diminished, suggesting the persistence of stress even after CHY removal. Confocal Laser Scanner Microscopy evidenced a diffuse aggregation and suffering of the treated cells, as a response to the stress encountered. In conclusion, the treatment with HY caused a stressing effect that persisted after its removal. The results suggest the potential of CHY application to control L. monocytogenes in food environments. [ABSTRACT FROM AUTHOR]

Endoglucanases are increasingly being touted as the ultimate solution for reducing energy consumption during the refining process in the pulp and paper industry. However, due to the high variety of endoglucanases in different enzyme formulations, these perform heterogeneously when applied to different pulps. In this study, the effect of four endoglucanases on softwood and hardwood pulp was studied using confocal laser scanning microscopy (CLSM) after addition of fluorescently labelled carbohydrate binding modules (CBMs). Nuclear magnetic resonance (NMR) analysis and high-performance liquid chromatography quantification of released oligo- and monosaccharides was performed for in-depth mechanistical investigation. Changes in the crystallinity levels caused by enzymatic degradation of amorphous regions were monitored by incubation with two different CBMs from Caldicellulosiruptor bescii and from Thermobifida fusca with high preference to either amorphous or crystalline regions of cellulose, respectively. When dosed at identical activity on the endoglucanase specific CellG5 substrate, CLSM analysis indicated the highest decrease of amorphous regions for those endoglucanases which were also most active in laboratory refining trials and which released highest amounts of cellooligomers from pulp. Using 13C-NMR analysis, an increase in para-crystalline cellulose caused by enzyme application was observed. Release of reducing sugars was determined at identical CellG5 dosage, indicating a high variance between the enzymes, especially when complex enzyme formulations were used. Scanning electron microscopy images were obtained for visualization of the endoglucanase activity. The results of mechanistical studies indicate that reduction of amorphous moieties of pulp by endoglucanases is especially beneficial for the refining process. [ABSTRACT FROM AUTHOR]

Introduction: Photodynamic therapy (PDT) is a treatment modality involving a dye that is activated by exposure to light of a specific wavelength in the presence of oxygen to form oxygen species causing localised damage to microorganisms. Aim: To determine the most effective bactericidal incubation and irradiation times of erythrosine-based PDT on in vivo-formed dental plaque biofilms. Methods: A randomised controlled study; 18-healthy adult participants wearing intraoral appliances with human enamel slabs to collect dental plaque samples in two separate periods of two weeks each for use in arm-1 and arm-2. These accumulated dental plaque samples were treated with PDT under different experimental conditions. Incubation times with photosensitiser (erythrosine) of 15 min and 2 min were used in arm-1 and arm-2, respectively, followed by light irradiation for either 15 min (continuous) or as a fractionated dose (5 × 30 sec). Following treatment, percentage reductions of total bacterial counts were compared between the different groups. In addition, confocal laser scanning microscopy (CLSM) and LIVE/DEAD® BacLight™ Bacterial Viability Kit were used to visualise the effect of PDT on in vivo-formed biofilms. Results: Significant reductions in the percentage of total bacterial counts (~93–95%) of in vivo-formed biofilms were found when using either 2 min or 15min incubation times and applying 15 min continuous light. Although when applying fractionated light, there was more cell death when 15 min incubation time was used (~ 91%) compared with the 2 min incubation time (~ 64%). CLSM results supported these findings. Conclusion: Improving the clinical usefulness of PDT by reducing its overall treatment time seems to be promising and effective in killing in vivo-formed dental plaque biofilms. [ABSTRACT FROM AUTHOR]

Background—The aim was to determine the influence of sonic activation in the tubular dentine penetration of bioceramic sealers. Methods—Forty mesiobuccal curved root canals of mandibular molars with an apical diameter smaller than #30 were prepared, divided into two groups, and filled with EndoSequence BC sealer, with or without sonic activation during its placement. Roots were sectioned at 3 mm, 6 mm, and 9 mm from the apex, producing a sample size of 120. The samples were evaluated using a confocal laser scanning microscope and comparing these images to the images obtained from an operatory microscope. The percentage of sealer penetration and maximum sealer penetration were evaluated. Statistical analysis was performed using the two-tailed Mann–Whitney U test, where statistical significance was set to p < 0.05. Results—Sonic activation showed higher values for the percentage of sealer penetration when compared at the 9 mm level (p = 0.03). A higher value of maximum sealer penetration was observed at all levels when the sealer was activated. Conclusions—The sonic activation of bioceramic cement resulted in higher sealer penetration into dentinal tubules. [ABSTRACT FROM AUTHOR]

Summary: The architecture of endosperm cell walls in Hordeum vulgare (barley) differs remarkably from that of other grass species and is affected by germination or malting. Here, the cell wall microstructure is investigated using (bio)chemical analyses, cryogenic scanning electron microscopy (cryo‐SEM) and confocal laser scanning microscopy (CLSM) as the main techniques. The relative proportions of β‐glucan, arabinoxylan and pectin in cell walls were 61, 34 and 5%, respectively. The average thickness of a single endosperm cell wall was 0.30 µm, as estimated by the cryo‐SEM analysis of barley seeds, which was reduced to 0.16 µm after malting. After fluorescent staining, 3D confocal multiphoton microscopy (multiphoton CLSM) imaging revealed the complex cell wall architecture. The endosperm cell wall is composed of a structure in which arabinoxylan and pectin are colocalized on the outside, with β‐glucan depositions on the inside. During germination, arabinoxylan and β‐glucan are hydrolysed, but unlike β‐glucan, arabinoxylan remains present in defined cell walls in malt. Integrating the results, an enhanced model for the endosperm cell walls in barley is proposed. Significance Statement: The carbohydrate composition and structure of barley endosperm cell walls are different from other grass species and change during malting/germination. Arabinoxylan, β‐glucan and pectin are distinguished microscopically, showing the colocalization of arabinoxylan and pectin. This is important for a better understanding of barley germination/malting and the brewing process. [ABSTRACT FROM AUTHOR]

Pholoe species described from northern European coastal waters are studied here, based on morphological and molecular methods. We confirm the current taxonomic status of P. assimilis , P. baltica , P. inornata and P. pallida. All species descriptions are reviewed and refined, barcodes are provided allowing the identification of these species based on molecular markers. Expanding morphological methods by using confocal laser scanning microscopy has added new characters to species diagnoses and makes species identification more secure. The reliability of characters traditionally used for identification of Pholoe species is evaluated. The newly presented identification key allows the delimitation of the four species reported for European coastal waters and also includes P. minuta and P. longa. The latter have been originally described from Greenland, but were often reported from European coastal waters. The results of our studies provide new insights into the distribution of the different species. Relationships of Pholoe species from mainly northern European waters are analysed based on molecular mitochondrial and nuclear markers. We stress the need for the prospective development of a precise terminology for prostomial appendages in Pholoe by including fine anatomical studies. [ABSTRACT FROM AUTHOR]

Planktonic diatoms thrive in the water column, and several species can transform into resting stages – spores or resting cells – that sink to the bottom of the sea. Resting stages are generally produced when environmental conditions are not optimal for growth and can remain viable in sediments for a long time. We tested different aspects related to the formation of spores in Chaetoceros socialis, one of the dominant diatoms in the Gulf of Naples (Mediterranean Sea, Italy). Observations in time lapse and confocal laser scanning microscopy allowed illustration of the formation of endogenous spores, in which valve deposition is accompanied by an acytokinetic nuclear division. The complete transition from vegetative cell to spore takes about 8–10 h and, if exposed to the light, spores can germinate again after a few days. The depletion of nitrogen source in the culture medium induced the formation of spores, with very similar dynamics among different sympatric strains; extremely high percentages of spores (up to >95%) were produced after 4–5 days. Once formed, spores can remain viable for up to nine months, and anoxic conditions favour their preservation. Identical dynamics of spore formation were detected in freshly established cultures and in cultures produced by the germination of spores kept dormant for different lengths of time. Our results suggest that spores of C. socialis can, in principle, rapidly shift between vegetative cells and resting stages that may explain the success of this species in coastal water. Our results also demonstrate that storage of resting spores may represent an alternative to cryopreservation. [ABSTRACT FROM AUTHOR]

[Display omitted] • A multi-emissive CDs-based fluorescence array combined with CLSM and LDA. • Synergistic recognition of bacteria and CDs at two pH levels and three excitations. • High discrimination performance for six foodborne bacteria with 100% accuracy. • Rapidly bacterial discrimination in mixture and food samples within a few minutes. The rapid discrimination of multiple foodborne pathogens is vital and challengeable in ensuring food safety. Herein, a facile fluorescence method was developed to distinguish different bacteria using multicolor-emissive carbon dots (CDs) based on confocal laser scanning microscopy (CLSM) and linear discriminant analysis (LDA). The array employed single CDs as receptors and indicators, two pH solutions and three excitation channels as sensing elements for multicolor imaging and recognition of multiple bacteria. Through one-step staining, the CDs were capable of rapid adsorbing onto all bacteria in 1 minute with varying affinities. The discrimination ability of the array was markedly enhanced by means of the synergistic recognition of bacteria and CDs across diverse pH levels and three excitation channels. By using LDA, six kinds of bacteria could be well discriminated with the classification accuracy of 100%. Meanwhile, the feasibility has been successfully expanded to mixture and food samples discrimination. [ABSTRACT FROM AUTHOR]

The aging behavior of thermoplastic polyurethane-modified asphalt (TPUMA) is intricate and the mechanisms of its thermal degradation are not well understood. This study examines the viscoelastic, chemical and morphological characteristics of thermoplastic polyurethane modified asphalt (TPUMA) under different aging conditions (RTFOT/PAV). The rheological master curve model of the S parameter was used to analyze the fitting effect on TPUMA, with SBS (styrene–butadiene–styrene) modified asphalt serving as the control group. The relationship between the phase angle plateau area and polymer aging degradation was elucidated. This study investigates the impact of polymer degradation on the high-temperature rheological behavior and medium-temperature fatigue properties of asphalt through temperature sweep, multiple stress creep recovery (MSCR), and linear amplitude sweep (LAS) tests. Fourier transform infrared spectrometer (FTIR) and confocal laser scanning microscopy (CLSM) are used to qualitatively and quantitatively analyze the chemical composition and morphological evolution of TPUMA during thermal-oxidative degradation. The findings suggest that TPUMA's performance changes in the aging environment is result from the dual coupling effect of polymer degradation and asphalt oxidative hardening. The closely arranged isocyanate (-NCO) and polar component molecular chains in asphalt slow down its oxidation speed, which enhances the aging and deformation resistance of TPUMA. The successful blending of TPU and asphalt was confirmed, and the degradation degree of the polymer was quantitatively characterized by the percentage of the degraded area. The results provide insights to better understand the interplay between TPU degradation and asphalt oxidation. [Display omitted] • Thermoplastic polyurethane (TPU) increases the complex modulus and rutting resistance of asphalt. • Jnr results reveal the dual coupling impact of polymer degradation and bitumen oxidative hardening on TPUMA rheology. • TPUMA underwent degradation and phase structure disruption between short-term and long-term aging. • The radar chart and correlation coefficient results show correlation between the macro and micro performance of TPUMA. [ABSTRACT FROM AUTHOR]

[Display omitted] Based on our previous report, the study was extended to investigate the impact of miconazole nitrate (MCN) loaded cationic/anionic nanoemulsions and nanoemulsion gels on permeation behaviour across artificial-membrane, EpiDerm, and rat skin. Nanoemulsions and gels were evaluated for size, charge, viscosity, size-distribution, pH, and percent entrapment efficiency (%EE). In vitro drug diffusion across artificial membrane and EpiDerm were conducted to get diffusion coefficients. Permeation profiles were studied using rat skin to investigate mechanistic insight of formulated mediated permeation followed by CLSM (confocal laser scanning microscopy), SEM (scanning electron microscopy), AFM (atomic force microscopy), and irritation studies. Results showed that MCNE11-Rh (probed cationic nanoemulsion at pH ∼ 7.2) and MNE11-Rh (probed anionic nanoemulsion at pH ∼ 7.2) showed size values of 158 nm and 145 nm, respectively whereas MCNE11-GR (probed cationic nanoemulsion gel at pH ∼ 6.8) and MNE11-GR (probed anionic nanoemulsion gel at pH ∼ 6.8) exhibited size values 257 nm and 243 nm, respectively. The %EE values were found to be as 91.5 % and 89.6 % for MCNE11-Rh and MNE11-Rh, respectively. The gels (∼6000 cP) elicited relatively high viscosity than nanoemulsions (∼3300 – 3500 cP). MCNE11-GR showed the highest values of permeation flux, diffusion rate, diffusion coefficient (D), and permeation coefficient (P) across artificial membrane, EpiDerm, and rat skin which may be attributed to three potential factors (cationic charge, composition, and hydration by the hydrophilic gel) working in tandem. Transepidermal water loss (TEWL) by the MCNE11-GR was maximum (14.4 g/m2h) than control (6.1 g/m2h) indicating augmented interaction of MCNE11-Rh with skin components. Conclusively, cationic nanoemulsion gel was promising carrier for enhanced permeation and the drug access to the dermal region to treat deep seated fungal infections. [ABSTRACT FROM AUTHOR]

A rapid, accurate, and sensitive reverse phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated for the estimation of Thymoquinone (TMQ) in API as well as in noisome. The chromatograms were developed with the mobile phase – water: 2-propanol: methanol (50:45:5 v/v/v) as a solvent system at 254 nm. The method was validated as per ICH guidelines for different parameters and the recovery of TMQ was calculated in developed niosomes. Further, TMQ loaded niosomes (TMQNIOS) were prepared and evaluated for different parameters. The optimized TMQNIOS (F3) was further evaluated for surface morphology, in vitro drug release, permeation study, and confocal laser scanning microscopic (CLSM) study. The method showed linearity range between 6.25 and 100 µg/ml with low detection limit and quantitation limit with a value of 2.08 and 6.25 µg/ml. The developed formulations showed the vesicle size and encapsulation efficiency in the range of 157.32 ± 3.15 to 211.44 ± 5.23 nm and 59.32 ± 4.87 to 83.21 ± 3.55%, respectively. The drug release result showed the significant higher release from TMQNIOS in compared to TMQDIS, and the release kinetics data showed Higuchi's equation with highest regression coefficient values. The permeation study and the confocal laser microscopy study further confirmed the enhancement in permeation of TMQ in the intestinal mucosa. [ABSTRACT FROM AUTHOR]

Dermal and transdermal drug therapy is increasing in importance nowadays in drug development. To completely utilize the potential of this administration route, it is necessary to optimize the drug release and skin penetration measurements. This review covers the most wellknown and up-to-date methods for evaluating the cutaneous penetration of drugs in vitro as a supporting tool for pharmaceutical research scientists in the early stage of drug development. The aim of this article is to present various experimental models used in dermal/transdermal research and summarize the novel knowledge about the main in vitro methods available to study skin penetration. These techniques are: Diffusion cell, skin-PAMPA, tape stripping, two-photon microscopy, confocal laser scanning microscopy, and confocal Raman microscopic method. [ABSTRACT FROM AUTHOR]

The susceptibility of four L. monocytogenes isolates from pork to sodium hypochlorite (SHY) and benzalkonium chloride (BZK) was tested. Minimum inhibitory concentration (MIC) values of 3500 ppm (SHY), or between 3 ppm and 13 ppm (BZK), were found. Minimum bactericidal concentration (MBC) values ranged from 3500 ppm to 4500 ppm (SHY), and from 3 ppm to 14 ppm (BZK). The effect of SHY and BZK on the architecture and cellular viability of 24-h-old biofilms formed by such strains on polystyrene was determined through confocal laser scanning microscopy (CLSM) in conjunction with fluorescent dyes for live cells (SYTO 9) and dead cells (propidium iodide). Strains were able to form biofilm (biovolume values in the observation field of 14,161 μm2 ranged between 103,928.3 ± 6730.2 μm3 and 276,030.9 ± 42,291.9 μm3). Treatment of biofilms for 10 min with SHY (1MIC or 1.5MIC) or BZK (0.5MIC, 1MIC or 1.5MIC) decreased the biovolume of live (potentially dangerous) cells. SHY reduced the cellular viability of biofilms by more than 90%. On the other hand, BZK was able to remove most biofilm mass (live and dead cells), but decreased cellular viability only to a lesser extent, this suggesting strong biofilm detachment and dissemination of live cells. • The susceptibility of four L. monocytogenes strains to two disinfectants was tested. • L. monocytogenes biofilms were exposed to disinfectants at 0.5MIC, 1MIC or 1.5MIC. • Most bacteria in biofilms were inactivated after contact with sodium hypochlorite. • Exposure of biofilms to benzalkonium chloride caused strong biofilm detachment. • Low doses of biocides could lead to the spread of live cells, a worrying finding. [ABSTRACT FROM AUTHOR]

A new vagrant eriophyoid mite species of the archaic genus Pentasetacus (Schliesske 1985), P. novozelandicusn. sp., is described with the aid of conventional microscopy, confocal laser scanning microscopy and scanning electron microscopy. It was found on Araucaria heterophylla, which is an araucarian that is endemic to Norfolk Island and introduced to New Zealand. Partial sequences of mitochondrial barcode COI gene and D1–D2 domains of nuclear rDNA of two pentasetacid mites, P. araucariae (MK903025 and MK898944) and P. novozelandicusn. sp. (MK903024 and MK898943) are provided. Molecular phylogenetic analyses of full-length D1–D2 eriophyoid sequences, including GenBank sequences and newly generated sequences of pentasetacids, confirmed the monophyly of Pentasetacidae but failed to resolve the basal phylogeny of Eriophyoidea. This may be because the D1–D2 domains of 28S are hypervariable in Eriophyoidea. Moreover, in pentasetacids D1–D2 sequences are about 20% shorter than in other eriophyoids, and thus harder to align. Two types of anal lobes are described in Eriophyoidea: (1) Eriophyidae s.l. and Phytoptidae s.l. have bilaterally symmetric lobes; (2) pentasetacids have non-divided lobes. The presence of an anal secretory apparatus, comprising internal structures that have previously been described in Eriophyidae s.l. and Phytoptidae s.l., is confirmed in pentasetacid genera. The phylogeny of pentasetacids is also discussed in the context of the paleobiography of Araucariaceae. [ABSTRACT FROM AUTHOR]

Knowledge on the detailed body plan in Palaeonemertea (a group putatively considered to retain ancestral character states within Nemertea) is essential in understanding the morphological evolution among not only nemerteans but also the rest of bilaterians in general; however, such information has been scanty in previous literature. In this study, we examined the body wall musculature, the position of the nervous system, and the musculature of the proboscis apparatus in 14 species of Archinemertea (a palaeonemertean clade comprised of Cephalotrichidae and Cephalotrichellidae) using confocal laser scanning microscopy with phalloidin labeling as well as classical Mallory's trichrome staining. Unexpectedly, all the archinemerteans possessed a diagonal muscle layer lying directly 'above'—instead of 'below', as usually found in most vermiform animals—the body wall outer circular muscle layer, a character state that has never been reported among Bilateria except for some acoels and proseriates. We speculate that the aberrant position of the outer diagonal muscle layer might have been actualized as a result of a flip-flop in a genetic regulatory circuit that controls the body wall muscular morphogenesis, which must have happened independently among the bilaterian evolution at least once in Acoela (Xenacoelomorpha), Proseriata (Platyhelminthes), and Archinemertea (Nemertea), respectively. Based on available phylogenetic data and the results from the present observation, we reconstructed the evolution of morphological characters within Archinemertea and its subclades. The outer diagonal muscle layer is likely a synapomorphy for Archinemertea (Cephalotrichidae + Cephalotrichellidae). Members of Balionemertes and Cephalotrichella possess some unique states for palaeonemerteans in muscular morphology (muscular strand associated with lateral nerve cord; longitudinal muscles between brain and extraganglionic tissue; proboscis with outer longitudinal muscle strands). We propose that muscle fibers embedded in lateral nerve cords, termed myofibrillae, in hoplo- and heteronemerteans are homologous with the body wall longitudinal muscles near the lateral nerves surrounded by circular muscles. [ABSTRACT FROM AUTHOR]

Abstract Regulatory developments regarding antifouling molecules encourage the search for non-toxic substances. Evaluation tools must evolve to highlight anti-adhesion effects rather than growth inhibition. The work presented here aimed at developing a method based on confocal laser scanning microscopy-microfluidic flow-cells in order to characterize microalgal biofilms. The first part of the work was dedicated to the setting-up of experimental parameters allowing the production of microalgal biofilms. The results obtained showed that it was indeed possible to produce reproducibly biofilms. The size of microalgal strains appeared to be a key-parameter in the adhesion rate and cells adhesion strength. Cylindrotheca closterium cells adhered in lower amount but formed denser biofilms than Porphyridium purpureum. The second part of the work focused on the evaluation of a known antifouling molecule, dibromohemibastadin-1 (DBHB). A comparison with the conventionally used method, multi-well plates experiments, was established. The multi-well plates experiments allowed the determination of minimum inhibitory concentration (MIC) for growth and adhesion inhibition (around 80 μM). The flow-cells combined with confocal laser scanning microscopy (CLSM) enabled the observation of biofilm, the determination of kinetics parameters of adhesion and an estimation of the adhesion strength. Highlights • Microalgal biofilms was grown in flow cell system and observed by CLSM. • Method able to evaluate antifouling compounds. • Method more informative than multi-well plate but complementary. [ABSTRACT FROM AUTHOR]

[Display omitted] • Proper precooking enhanced the extensibility of autoclaved recooked noodles (ARNs). • Autoclaving and recooking processes increased the tightly bound water proportion. • Autoclaving and recooking processes further strengthened the gluten network of ARNs. • "AngioTool" analysis of CLSM proved that a strengthened gluten network appeared. To improve the quality of autoclaved recooked noodles (ARNs), this study explored the effects of precooking on the sensory and tensile properties of ARNs from the perspectives of changes in protein structure and water distribution. The results showed that the ARNs of two kinds of pretreatments (Boiling 2 min, Boiling 1 min + Steaming 2 min) presented the best sensory quality (average score ≥ 7.50) and high tensile properties (tensile distance ≥ 45.24 mm). After autoclaving and recooking, the proportion of tightly bound water increased by 11.30%–12.52%, resulting in stronger water–solid interaction. The results of laser confocal microscopy (CLSM) proved that a strengthened gluten network (protein percentage area ≥ 40.28%; junction density ≥ 10.96 × 10−4) appeared. Therefore, appropriate precooking treatment could effectively improve the sensory quality and tensile properties of ARNs by enhancing the tightly bound water ratio and strengthening the gluten network. [ABSTRACT FROM AUTHOR]

Τhis study investigates the ability of whey proteins, which were pre-treated with ethanol (0–70%) and subjected tothermal treatment (70 °C for 60 min), to form thermo-reversible gels after removing the alcohol through freeze-drying. The ethanol-free gels were monitored by means of rheology and confocal laser scanning microscopy. Moreover, the role of ethanol during heating of whey proteins was investigated by UV difference spectroscopy. The results showed a retention of the denatured character (30–35%) of whey proteins after removal of ethanol, regardless of whether heating was applied. Intermolecular disulfide connections increased in whey protein solutions pretreated with 30 and 50% w/w ethanol. The determination of free –SH groups demonstrated that their number was increased by heating in the presence of ethanol and after ethanol's removal. Dynamic rheological measurements showed that ethanol-free gels, previously heated with 50% w/w ethanol, formed thermo-reversible gels upon cooling to 5 °C and exhibited significantly higher storage modulus values (G'∼300 Pa) than the counterparts without pretreatment of the proteins with ethanol (G' = 0.0002 Pa). Generally, the partial retention of the denaturing effect of ethanol on whey proteins after its removal and their ability to form thermo-reversible gels could lead to new applications in the food and pharmaceutical industries. [Display omitted] • Ethanol-free thermo-reversible whey protein (WPI) gels were obtained. • Ethanol exerted a residual partial denaturing effect after its removal. • Pretreatment with combined heating and ethanol led to increased free –SH groups. • Ethanol-free WPI gels had modified properties than native WPI gels. [ABSTRACT FROM AUTHOR]

In this study, the dehydration kinetics, status of water and oil distribution of microwave-assisted vacuum frying (MVF) potato chips were analyzed combining with NMR and confocal laser scanning microscopy (CLSM). Results showed that the MVF markedly increased the moisture evaporation kinetics and effective moisture diffusivity compared to vacuum frying (VF). The higher microwave power level (1000 W) used achieved higher moisture evaporation rate and higher effective moisture diffusivity, The Logarithmic model exhibited the best fit for the obtained data of MR versus frying time in the MVF. The NMR analysis showed the free water in the samples firstly evaporate and the linkage between water and structure of materials becomes tighter with the frying time. The total oil content, surface and structural oil content were all significantly lower in MVF samples than that in VF samples. The CLSM analysis confirmed that less oil adhere to the surface and less oil trapped in the structure in MVF slices. The surface morphology showed that there were less ruptured and damaged cells in MVF samples and helped to explain the reduction of oil content. [ABSTRACT FROM AUTHOR]

Phototrophic microorganisms are very abundant in extreme environments, where are subjected to frequent and strong changes in environmental parameters. Nevertheless, little is known about the physiological effects of these changing environmental conditions on viability of these microorganisms, which are difficult to grow in solid media and have the tendency to form aggregates. For that reason, it is essential to develop methodologies that provide data in short time consuming, in vivo and with minimal manipulating the samples, in response to distinct stress conditions. In this paper, we present a novel method using Confocal Laser Scanning Microscopy and a Dual Laser (CLSM-DL) for determining the cell viability of phototrophic microorganisms without the need of either staining or additional use of image treating software. In order to differentiate viable and nonviable Scenedesmus sp. DE2009 cells, a sequential scan in two different channels was carried out from each same xyz optical section. On the one hand, photosynthetic pigments fluorescence signal (living cells) was recorded at the red channel (625- to 785-nm fluorescence emission) exciting the samples with a 561-nm laser diode, and an acousto-optic tunable filter (AOTF) of 20%. On the other hand, nonphotosynthetic autofluorescence signal (dead cells) was recorded at the green channel (500- to 585-nm fluorescence emission) using a 405-nm UV laser, an AOTF of 15%. Both types of fluorescence signatures were captured with a hybrid detector. The validation of the CLSM-DL method was performed with SYTOX green fluorochrome and electron microscopic techniques, and it was also applied for studying the response of distinct light intensities, salinity doses and exposure times on a consortium of Scenedesmus sp. DE2009. [ABSTRACT FROM AUTHOR]

The isothermal and non-isothermal crystallization behavior of blast furnace (BF) slag with varying CaO/SiO 2 ratios was in-situ observed through a confocal scanning laser microscopy (CLSM). The crystallization kinetics was analyzed and precipitation of crystalline phases was discussed by theoretical analysis from FactSage 8.0 and X-ray Diffraction (XRD). The results showed that, with the increasing CaO/SiO 2 ratio from 1.0 to 1.3, the crystals morphology changed first from rectangles and squares to small-size and irregular, and further to long strip shape at the cooling rate of 100 °C/min. The onset crystallization temperature in CTT diagram was higher at a larger CaO/SiO 2 ratio due to the decrease of polymerization degree of the slag. For the isothermal crystallization process at 1250°C, the crystals morphology changed gradually from lamellar to long-trip or columnar, and the incubation period in TTT diagram became shorter with the increase of CaO/SiO 2 ratio. Avrami index n decreased from appropriately 2.3 to 1.5, and the growth mode of crystals converted from two-dimensional to one- dimensional. XRD analysis showed that only merwinite existed in four slags at 1380°C. Akermanite and gehlenite were generated at 1350°C, and their amount first increased greatly and then decreased slightly with a continuous increase of CaO/SiO 2 ratio. At the low temperature, the number of akermanite and gehlenite reduced, while that of merwinite increased obviously in the slags when CaO/SiO 2 ratio was 1.2 and 1.3, indicating that increasing CaO/SiO 2 ratio promoted the generation of merwinite. [ABSTRACT FROM AUTHOR]

Confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were used to investigate the microstructural changes in wheat ( Triticum aestivum L.) during the malting process. With the help of these powerful optical tools, the organization of starch, proteins and cell walls of cereals could be investigated separately in projections and the microstructure was visualized with an imaging software. Both microscopes were necessary for the investigation and the evaluation of microstructural changes, because SEM and CLSM have different merits and optical benefits. CLSM achieves an overview of single compounds of the grain, which can be used to quantify them. On the other hand, SEM has a higher magnification, where a deeper insight in the kernel structure could be obtained. In particular, the surface of starch kernels can be visualized, when they get degraded by enzymes. Furthermore, a detailed starch structure can be monitored by using SEM. Various grain regions, such as aleurone layer, starchy endosperm and germ, were visualized with the CLSM, and differences in the single fractions were clearly visible. Starch in unmalted cereals revealed that it is embedded in a compact protein network, which consists of protein bodies and is degraded during the malting process. Also the cell walls, mainly consisting of β-glucan, are clearly visible in the unmalted stage and get more and more degraded during the malting process. Hence the differences in the overall structure of unmalted to malted grains were investigated by CLSM and SEM and verified using established malt analysis. [ABSTRACT FROM AUTHOR]

In situ 3D characterization of a filtration cake obtained with a model bidisperse suspension containing two sizes of fluorescent particles (1µm and 4.8µm) has been performed using Confocal Laser Scanning Microscopy (CLSM). The suspensions were dead-end filtered through 0.8µm pore diameters silicon nitride microsieves, under a constant flow rate. The membrane was put in a specific filtration chamber previously designed [1], allowing direct on-line microscopic observations of particles deposition. The final position of the first particles arriving on the membrane was investigated. Then, the online deposition of the following particles was recorded and, thus, the cake build-up could be analyzed layer by layer. In addition, to analyze the protective effect of a layer of large particles, 1µm particles were filtrated over a previously deposited cake formed with the 4.8µm microspheres. [ABSTRACT FROM AUTHOR]

Confocal laser scanning microscopy (CLSM) is often used to evaluate biofilm development or biofouling mitigation in membrane systems. However, several methods of CLSM sample preparation exist. In this paper, we evaluate the effects of three preparation techniques - dry, confined (wet), and unconfined (immersed) mounting - on CLSM-derived biofilm architecture and dimensions. Although placing a wet or dry biofilm between a slide and a coverslip before viewing is relatively common, our results show that this confinement significantly alters the biofilm observed. Therefore, biofilms should be viewed in an unconfined and hydrated state that allows for full extension of the biofilm structure in a media-filled viewing well of fixed depth (~250μm). Pseudomonas aeruginosa biofilms were grown on thin-film composite reverse osmosis membranes and glass coupons. Dry and confined mounting of 24 and 48h biofilms resulted in biofilms with low 3-D complexity and thickness (14 and 18μm, respectively). Measured biofilm thickness was significantly higher on samples prepared using unconfined mounting (55μm). Additionally, the reduction in biofilm thickness and biovolume observed after treatment with biocidal compounds was significantly less on the dry and confined biofilms than the unconfined samples. Our results strongly suggest that biofilms on membranes be prepared for microscopy using unconfined mounting to accurately assess biofilm structure and dimensions. Unconfined mounting will allow for accurate CLSM assessment of membrane biofilm structure, dimensions, and biofouling mitigation measures in membrane systems. [ABSTRACT FROM AUTHOR]

The mechanism commonly assumed to explain the exceptional lift of laminated doughs is that fat layers act as a barrier to steam produced during baking. Successive folding and sheeting steps increase the number of fat layers and the lift, up to a point (48 fat layers in our study) where fat layers would suddenly rupture. Void spaces in the fat layers were characterized on confocal laser scanning microscopy images of Danish pastry from 8 to 64 fat layers: the greater the number of fat layers, the greater the average number of void spaces, following a power law from 16 layers. At 64 fat layers, Danish pastry appeared to be over-laminated. The size of small void spaces (2-50μm) was shown to be independent of the number of fat layers. The number of large void spaces was linearly dependent on the number of fat layers. These results failed to show any transition at 48 fat layers. [ABSTRACT FROM AUTHOR]

The nereidid annelid Platynereis dumeriliiemerged as a well-understood model organism. P dumerilii and P. massiliensis are sister taxa, which are morphologically indistinguishable as adults. Interestingly, they exhibit highly contrasting life-histories: while Pdumeriliiis a gonochorostic species with planktonic feeding larvae, P massiliensis is a protandric hermaphrodite with lecitotrophic semi-direct-development in brood tubes. Using light microscopy and immunohistochemical methods coupled with confocal laser scanning microscopy, we describe the development of P massiliensis. Musculature was stained with phalloidin-rhodamine. FMRFamide, acetylated a-tubulin, and serotonin were targeted by antibodies for the staining of neuronal structures. Additionally, eye development was investigated with the specific 22C10-antibody.The development of P massiliensis is characterized by the absence of a free-swimming stage, a late development of food uptake, and the presence of a large amount of yolk even in late juvenile stages. Most notably, early juvenile stages already exhibit an organization of several organ systems that resembles those of adults. Larval characters present in the free-swimming feeding larvae of P dumerilii, as e.g. the apical organ and larval eyes, are absent and regarded to be lost in developing stages of P. massiliensis. Many of the differences found in the development of these two species can be described in the context of heterochronic changes. We strongly advocate expanding evolutionary developmental studies from the well-established model annelid P. dumerilii to the closely related R massiliensis to study the evolutionary conservation and divergence of genetic pathways involved in developmental processes. [ABSTRACT FROM AUTHOR]

Abstract: Lipid oxidation is a widespread phenomenon in foods and other systems of biological origin. Detection methods for early stages of lipid oxidation are in demand to understand the progress of oxidation in space and time. The fluorescence spectrum of the nonpolar fluorescent probe BODIPY665/676 changes upon reacting with peroxyl radicals originating from 2,2′-azobis(2,4-dimethyl)valeronitrile and tert-butoxyl radicals generated from di-tert-butylperoxide. The excitation wavelength of the main peak of BODIPY665/676 was 675nm in the fluorometer, and 670nm under the microscope, and the optimum excitation wavelength for the secondary peak of BODIPY665/676 was 580nm. Advantages of using BODIPY665/676 are fewer problems with autofluorescence and the possibility of combining several fluorescent probes that are excited and emitted at lower wavelengths. However, because of the spectrum of the probe, specific lasers and detectors are needed for optimal imaging under the microscope. Furthermore, BODIPY665/676 is resistant to photobleaching at both excitation wavelengths, 670 and 580nm. In diffusion studies, BODIPY665/676 is highly lipophilic, remaining in the lipid phase and not diffusing into the aqueous phase or between lipid droplets. [Copyright &y& Elsevier]

Abstract: Saprolegnia spp. can cause mortality and economic losses in freshwater fish and eggs. Biofilm formation is generally regarded as a virulence factor, and biofilms can be an important cause of infection recurrence. Evidence of persistent sources of Saprolegnia infections on fish and eggs in fish farms support the assumption that Saprolegnia spp. might be able to form biofilms. In this study, we aimed to test the ability of Saprolegnia to form biofilms where it can survive, reproduce and resist different chemicals used for its control. Naturally formed biofilms were obtained from laboratory aquaria. Saprolegnia growth within these biofilms was demonstrated with light microscopy and confirmed by isolation. Isolates were identified morphologically and molecularly on the basis of ITS-sequences. Two isolates were identified as Saprolegnia parasitica, a species known to be highly pathogenic for fish, while the other belonged to S. australis. Selected Saprolegnia strains obtained from natural biofilms were then used to establish simple methods for in vitro induction of Saprolegnia biofilm. The ability of Saprolegnia isolates to form biofilms with subsequent production of infective motile zoospores within the biofilm was documented by light and confocal laser scanning microscopy. We demonstrate for the first time that isolates of S. parasitica and S. australis can form biofilm communities together with multiple microorganisms, wherein they grow and reproduce. It is therefore likely that natural biofilms constitute incessant Saprolegnia reservoirs in nature and aquaculture. [Copyright &y& Elsevier]

Background: Periodontitis is caused by a highly complex consortium of bacteria that establishes as biofilms in subgingival pockets. It is a disease that occurs worldwide and its consequences are a major health concern. Investigations in situ are not possible and the bacterial community varies greatly between patients and even within different loci. Due to the high complexity of the consortium and the availability of samples, a clear definition of the pathogenic bacteria and their mechanisms of pathogenicity are still not available. In the current study we addressed the need of a defined model system by advancing our previously described subgingival biofilm model towards a bacterial composition that reflects the one observed in diseased sites of patients and analysed the structure of these biofilms. Results: We further developed the growth media by systematic variation of key components resulting in improved stability and the firm establishment of spirochetes in the 10-species subgingival Zurich biofilm model. A high concentration of heat-inactivated human serum allowed the best proliferation of the used species. Therefore we further investigated these biofilms by analysing their structure by confocal laser scanning microscopy following fluorescence in situ hybridisation. The species showed mutual interactions as expected from other studies. The abundances of all organisms present in this model were determined by microscopic counting following species-specific identification by both fluorescence in situ hybridisation and immunofluorescence. The newly integrated treponemes were the most abundant organisms. Conclusions: The use of 50% of heat-inactivated human serum used in the improved growth medium resulted in significantly thicker and more stable biofilms, and the quantitative representation of the used species represents the in vivo community of periodontitis patients much closer than in biofilms grown in the two media with less or no human serum. The appearance of T. denticola, P. gingivalis, and T. forsythia in the top layer of the biofilms, and the high abundance of T. denticola, reflects well the microbial situation observed at diseased sites. The improved model biofilms will allow further investigations of interactions between individual species and of the effects of atmospheric or nutritional changes, as well as interactions with tissue cells. [ABSTRACT FROM AUTHOR]

Sepsis is characterized by a systemic inflammatory response caused by infection, and can result in organ failure and death. Removal of inflammatory mediators such as cytokines from the circulating blood is a promising treatment for severe sepsis. We are developing an extracorporeal hemoadsorption device to remove cytokines from the blood using biocompatible, polymer sorbent beads. In this study, we used confocal laser scanning microscopy (CLSM) to directly examine adsorption dynamics of a cytokine (IL‐6) within hemoadsorption beads. Fluorescently labeled IL‐6 was incubated with sorbent particles, and CLSM was used to quantify spatial adsorption profiles of IL‐6 within the sorbent matrix. IL‐6 adsorption was limited to the outer 15 μm of the sorbent particle over a relevant clinical time period, and intraparticle adsorption dynamics was modeled using classical adsorption/diffusion mechanisms. A single model parameter, α = qmaxK/D, was estimated by fitting CLSM intensity profiles to our mathematical model, where qmax and K are Langmuir adsorption isotherm parameters, and D is the effective diffusion coefficient of IL‐6 within the sorbent matrix. Given the large diameter of our sorbent beads (450 μm), less than 20% of available sorbent surface area participates in cytokine adsorption. Development of smaller beads may accelerate cytokine adsorption by maximizing available surface area per bead mass. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010 [ABSTRACT FROM AUTHOR]