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Mast cells are an important component of connective tissue and can be selectively visualized with basic aniline dyes. One of them is cresyl violet, which stands out among others due to its capacity for fluorescence. The aim of this study is to optimize the histochemical method of cresyl violet staining for the selective detection of mast cells in rat myocardium. In this work, various staining protocols are presented for transmitted light microscopy of the visible range and for confocal microscopy. It is shown that the highest degree of staining selectivity can be achieved by differentiating the staining with hydrochloric acid. High color contrast, achieved due to pronounced metachromasia, between mast cells and surrounding tissue elements can be achieved using an aqueous mounting medium. Fluorescent cresyl violet staining in combination with confocal microscopy allows a clearer visualization of mast cell granules and the study of their structure. [ABSTRACT FROM AUTHOR]

Main conclusion: During evolution, similar vectors of adaptive radiation may have evolved in the subtribe Malaxidinae. This was manifested in homologous series of variability in suspensor shape and seed coat ultrasculpture in clades of terrestrial and epiphytic orchids. The present study examines the variability of embryonic traits across clades and subclades of subtribe Malaxidinae (Orchidaceae), previously identified by molecular genetic data. Ovules and seeds from fruits of orchids of the genera Crepidium, Liparis (sections Cestichis and Blepharoglossum), Dienia, and Oberonia were examined by confocal laser microscopy with fluorescent dye staining. The branched or rounded suspensor in the studied species was unicellular and originated from the nondividing basal cell cb. The elongated or flattened transmission structure of one or two cells was located at the junction of the suspensor and the embryo proper. Two species (Oberonia gammiei and Liparis elliptica) were found to have unitegmal ovules. Three morphological groups of seeds were identified based on the shape and sculpture of the periclinal cell wall. A comparative analysis of the embryological characters in Malaxidinae species reveals that the lobed suspensor is a homoplasy present in different subclades of terrestrial and epiphytic orchids. The flat transmission cell is an apomorphy in the Cestichis subclade. Similarly, the independent formation of the unitegmal ovule occurred in two subclades of epiphytic orchids. The results of our study suggest that similar adaptive radiation vectors may have evolved in the subtribe Malaxidinae in the orchids we studied. [ABSTRACT FROM AUTHOR]

Background Study Design Endpoints Patients and Methods Radical prostatectomy (RP) represents the cornerstone of surgical treatment for prostate cancer. Assessing surgical margin status intraoperatively with current techniques remains challenging due to high costs in the context of an already stretched pathology workforce. Fluorescence confocal microscopy (FCM) is a promising technique to detect margins in prostate cancer surgery not bound by such limitations.The Imperial Prostate 8 – Fluorescence Confocal Microscopy for Rapid Evaluation of Surgical Cancer Excision (IP8‐FLUORESCE) study is a multicentre, prospective, ex vivo, ‘blinded’, comparative cohort study. It aims to assess the accuracy of digital FCM for detection of prostate cancer at surgical margins compared to traditional histopathology.The primary endpoint is the accuracy of digital FCM for detection of prostate cancer at surgical margins on a per‐patient level, reported with sensitivity, specificity, positive and negative predictive values.A total of 153 patients with localised prostate cancer undergoing robot‐assisted RP across three UK National Health Service tertiary referral centres will be recruited. Following RP, prostate specimens will undergo immediate immersion in Acridine Orange solution, scanning ‘en face’ with FCM using the Histolog® Scanner, and subsequent formalin fixation and paraffin embedding. Two independent, ‘blinded’ uro‐pathologists will report both the FCM images and the histopathology slides. Recruitment commenced on 17 August 2023. [ABSTRACT FROM AUTHOR]

Intraoperative frozen section (IFS) is used with the intention to improve functional and oncological outcomes for patients undergoing radical prostatectomy (RP). High resource requirements of IFS techniques such as NeuroSAFE may preclude widespread adoption, even if there are benefits to patients. Recent advances in fresh‐tissue microscopic digital imaging technologies may offer an attractive alternative, and there is a growing body of evidence regarding these technologies. In this narrative review, we discuss some of the familiar limitations of IFS and compare these to the attractive counterpoints of modern digital imaging technologies such as the speed and ease of image generation, the locality of equipment within (or near) the operating room, the ability to maintain tissue integrity, and digital transfer of images. Confocal laser microscopy (CLM) is the modality most frequently reported in the literature for margin assessment during RP. We discuss several imitations and obstacles to widespread dissemination of digital imaging technologies. Among these, we consider how the 'en‐face' margin perspective will challenge urologists and pathologists to understand afresh the meaning of positive margin significance. As a part of this, discussions on how to describe, categorize, react to, and evaluate these technologies are needed to improve patient outcomes. Limitations of this review include its narrative structure and that the evidence base in this field is relatively immature but developing at pace. [ABSTRACT FROM AUTHOR]

Nanocomposite alginate hydrogel containing Propranolol hydrochloride (PNL) cerosomes (CERs) was prepared as a repurposed remedy for topical skin Methicillin-Resistant Staphylococcus aureus (MRSA) infection. CERs were formed via an ethanol injection technique using different ceramides, Kolliphores® as a surfactant, and Didodecyldimethylammonium bromide (DDAB) as a positive charge inducer. CERs were optimized utilizing 13. 22 mixed-factorial design employing Design-Expert® software, the assessed responses were entrapment efficiency (EE%), particle size (PS), and zeta potential (ZP). The optimum CER, composed of 5 mg DDAB, ceramide VI, and Kolliphor® RH40 showed tubular vesicles with EE% of 92.91 ± 0.98%, PS of 388.75 ± 18.99 nm, PDI of 0.363 ± 0.01, and ZP of 30.36 ± 0.69 mV. Also, it remained stable for 90 days and manifested great mucoadhesive aspects. The optimum CER was incorporated into calcium alginate to prepare nanocomposite hydrogel. The ex-vivo evaluation illustrated that PNL was permeated in a more prolonged pattern from PNL-loaded CERs nanocomposite related to PNL-composite, optimum CER, and PNL solution. Confocal laser scanning microscopy revealed a perfect accumulation of fluorescein-labeled CERs in the skin. The in-silico investigation illustrated that the PNL was stable when mixed with other ingredients in the CERs and confirmed that PNL is a promising candidate for curing MRSA. Moreover, the PNL-loaded CERs nanocomposite revealed superiority over the PNL solution in inhibiting biofilm formation and eradication. The PNL-loaded CERs nanocomposite showed superiority over the PNL-composite for treating MRSA infection in the in-vivo mice model. Histopathological studies revealed the safety of the tested formulations. In conclusion, PNL-loaded CERs nanocomposite provided a promising, safe cure for MRSA bacterial skin infection. [ABSTRACT FROM AUTHOR]

Respiratory diseases in ruminants are responsible for enormous economic losses for the dairy and meat industry. The main causative bacterial agent of pneumonia in ovine is Mannheimia haemolytica A2. Due to the impact of this disease, the effect of the antimicrobial protein, bovine lactoferrin (bLf), against virulence factors of this bacterium has been studied. However, its effect on biofilm formation has not been reported. In this work, we evaluated the effect on different stages of the biofilm. Our results reveal a decrease in biofilm formation when bacteria were pre-incubated with bLf. However, when bLf was added at the start of biofilm formation and on mature biofilm, an increase was observed, which was visualized by greater bacterial aggregation and secretion of biofilm matrix components. Additionally, through SDS-PAGE, a remarkable band of ~80 kDa was observed when bLf was added to biofilms. Therefore, the presence of bLf on the biofilm was determined through the Western blot and Microscopy techniques. Finally, by using Live/Dead staining, we observed that most of the bacteria in a biofilm with bLf were not viable. In addition, bLf affects the formation of a new biofilm cycle. In conclusion, bLf binds to the biofilm of M. haemolytica A2 and affects the viability of bacteria and the formation a new biofilm cycle. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to evaluate the surface properties and bacterial adhesion of computer-aided design-computer-aided manufacturing (CADCAM) restorative materials. Methods: Four CAD-CAM resin-based blocks (Vita Enamic, Shofu block HC, Cerasmart [CS] and Lava Ultimate [LU]) and a leucite-reinforced glass ceramic block (IPS Empress CAD) were used in the present study. Specimens prepared with dimensions of 10 × 10 × 1 mm were polished. Surface characteristics were assessed with hydrophobicity and surface free energy (SFE) analysis. Surface roughness was measured using a profilometer, and elemental and topographic evaluations were performed with SEM-EDX analysis. After being kept in artificial saliva for 1 h, Streptococcus mutans (S. mutans) and Streptococcus mitis (S. mitis) were incubated separately in 5% CO2 atmosphere at 37°C for 24 h. The adhered bacteria were counted as ×108 CFU/mL. Results: Surface roughness, contact angle and SFE measurement values were found to be in the range of 0.144-0.264 Ra, 28.362°-70.074° and 39.65-63.62 mN/m, respectively. The highest adhered amount of S. mutans was found in CS and the lowest in LU, while there was no significant difference between the amounts of adhered S. mitis. Conclusion: Despite differences in the surface properties of the materials used for the study, the materials exhibited identical properties with respect to bacterial adhesion. [ABSTRACT FROM AUTHOR]

Microbial exopolysaccharides (EPS) are extracellular polymeric substances synthesized by bacteria, fungi or algae which are now one of the important food ingredients. In this study, glucan EPS was isolated from lactic acid bacteria (LAB) of fermented food origin and was incorporated in yoghurt to enhance its techno-functional attributes and rheological properties. Glucan EPS concentration at 1% showed significant improvement in syneresis, wheying off, water-holding capacity, and sensorial properties of yoghurt. The water holding capacity (WHC) increased from 47% to about 63% while syneresis decreased from 52% to nearly 30%. Rheological features such as viscosity, storage modulus, loss modulus, and firmness were significantly higher for 1% EPS containing yoghurt (1% ECY) than that of the control. The average particle size of 1% ECY was increased from 2.35 µm to 3.46 µm. The microstructure of EPS containing yoghurt was studied by scanning electron microscopy and confocal laser scanning microscopy. EPS supports the growth of probiotic microflora of yoghurt, which was revealed by analyzing the microbial count of yoghurt. The antioxidant activity as 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (mg Trolox/100 g sample) and reducing power assay (mg ascorbic acid/100 g sample) for 1% ECY and control yoghurt were found to be 27.9 (mg Trolox/100 g sample), 6.44 (mg Trolox/100 g sample) and 0.25 (mg ascorbic acid/100 g sample), 0.19 (mg ascorbic acid/100 g sample), respectively. Significant enhancement of these bio-functional attributes of EPS containing yoghurt makes the glucan EPS a suitable hydrocolloid to develop various functional foods. [ABSTRACT FROM AUTHOR]

The aim of current study was to prepare and optimize the nanostructured lipid carrier (NLC) for transdermal gliclazide (GCZ) delivery. The melt emulsification followed by ultrasonication technique was utilized to prepare GCZ–loaded-NLC. The optimization of NLC was achieved by Box-Behnken design. The surfactant (Tween 20) % (X1), total lipid (glyceryl monostearate + Lauroglycol™ 90) % (X2), and sonication time (min) (X3) were chosen as independent variables, while particles size (Y1), polydispersity index (Y2), and entrapment efficiency (Y3) were selected as response. Further, the prepared NLC was estimated for in vitro drug release study, ex vivo skin permeation and confocal laser scanning microscopy (CLSM) studies. Results of present study demonstrated that the optimized GCZ loaded NLC formulation presented the particles size, polydispersity index, zeta potential, and entrapment efficiency of 120.4 nm, 0.316, −5.58 mV, and 87.32% respectively. The in vitro drug release study indicated drug release of 68.27 ± 2.98% and 45.87 ± 2.85% for GCZ-NLC and conventional GCZ dispersion respectively. The GCZ-NLC-Gel formulation exhibited transdermal drug permeation of 76.89 ± 2.52% as compared to conventional gel (45.65 ± 2.79%). The CLSM of rat skin treated with NLC gel exhibited a deeper permeation (35 μm) in comparison to the conventional gel (15 μm). In conclusion, GCZ loaded NLC was successfully optimized using Box-Behnken design that contributed many advantages over conventional formulation and thus demonstrating NLC as propitious carriers for the transdermal GCZ delivery. [ABSTRACT FROM AUTHOR]

Copyright of Journal der Deutschen Dermatologischen Gesellschaft is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Summary: Actinic keratosis (AK) is considered a chronic and recurring in situ skin neoplasia, with a possible transformation into invasive squamous cell carcinoma (SCC). Among others, predominant risk factors for development of AK are UV‐light exposure and immunosuppression. Basal epidermal keratinocyte atypia (AK I) and proliferation (PRO score) seem to drive malignant transformation, rather than clinical appearance of AK (Olsen I–III). Due to the invasiveness of punch biopsy, those histological criteria are not regularly assessed. Non‐invasive imaging techniques, such as optical coherence tomography (OCT), reflectance confocal microscopy (RCM) and line‐field confocal OCT (LC‐OCT) are helpful to distinguish complex cases of AK, Bowen's disease, and SCC. Moreover, LC‐OCT can visualize the epidermis and the papillary dermis at cellular resolution, allowing real‐time PRO score assessment. The decision‐making for implementation of therapy is still based on clinical risk factors, ranging from lesion‐ to field‐targeted and ablative to non‐ablative regimens, but in approximately 85% of the cases a recurrence of AK can be observed after a 1‐year follow‐up. The possible beneficial use of imaging techniques for a non‐invasive follow‐up of AK to detect recurrence or invasive progression early on should be subject to critical evaluation in further studies. [ABSTRACT FROM AUTHOR]

Background: Reflectance confocal microscopy (RCM) has a defined in vivo morphology of vitiligo and re-pigmentation. Combination therapies seem more effective than monotherapies. Objective: We aim to describe the clinical and RCM features of re-pigmentation with combined narrowband ultraviolet B (NB-UVB) and piperine-based topical treatment in localized vitiligo. Methods: Eight patients enrolled at a single center received combined treatment: topical treatment was applied twice daily + NB-UVB twice weekly for 2 × 2-month periods. Clinical changes were analyzed by the Vitiligo Noticeability Scale (VNS) and percentage of re-pigmentation. The evaluator agreement was assessed. Predefined RCM features had the presence/absence of (i) blood vessels, (ii) dendritic cells, and the quantity of (i) an irregular honeycombed pattern and (ii) non-pigmented papillae. Clinical and RCM monitoring was performed at the baseline, 2, 3, 5, and 7 months. Results: Macules were "slightly less noticeable" with 25–50% re-pigmentation. Irregular honeycomb patterns and non-pigmented papillae were significantly less frequently observed, and in less extended areas (T1 vs. T2, p = 0.039; T0 vs. T1, p = 0.005 and T2 vs. T4, p = 0.033). Dendritic cells and blood vessels improved, with significant changes in blood vessels (T1 vs. T2, p = 0.005 and T3 vs. T4, p = 0.008). Conclusions: RCM confirms the morphological changes induced by combined treatment for localized vitiligo. [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]

An ideal anti‐counterfeiting technology is desired to be unclonable, nondestructive, mass‐producible, and accompanied with fast and robust authentication under various external influences. Although multiple anti‐counterfeiting technologies have been reported, few meet all of the above‐mentioned features. Herein, a mechanically driven patterning process is reported to produce higher dimensional Ti3C2Tx MXene topographies in a scalable yet unclonable manner, which can be used as anti‐counterfeiting tags. By using a high‐speed confocal laser microscopy, the complex topographies can be extracted within one minute and then reconstructed into 3D physical unclonable function (PUF) keys. Meanwhile, a Siamese neural network model and a feature‐tracking software are built to achieve a pick‐and‐check strategy, enabling highly accurate, robust, disturbance‐insensitive tag authentication in practical exploitations. The 3D PUF key‐based anti‐counterfeiting technology features with several advances, including ultrahigh encoding capacities (≈10144 000‐107 800 000), fast processing times (<1 min), and high authentication accuracy under various external disturbances, including tag rotations (≈0°‒360°), tag dislocation(s) in x(y) directions (≈0%‒100%), tag shifts in z‐direction (≈0%‒28%), tag tilts (≈0°‒5°), differences in contrasts (20%‒60%) and laser power (6.0‒9.0 µW). The anti‐counterfeiting technology promises information security, encoding capacity, and authentication efficiency for the manufacturer‐distributor‐customer distribution processes. [ABSTRACT FROM AUTHOR]

Simple Summary: The detection of thyroid nodules is increasing worldwide. Fine needle aspiration biopsy (FNA) is used to differentiate benign and malignant nodules and to address patient management; about 30% of patients receive indeterminate or inadequate cytological report. This clinical problem needs an innovative approach, overcoming the limits of traditional cytological diagnostics. Fluorescence laser confocal microscopes (FCM) is a new optical technique for allowing immediate digital imaging of fresh unfixed tissues, and it recently obtained attention for real-time assessment of sample adequacy and diagnostic evaluation for small biopsies and cytological samples. Herein we tested FCM technology for evaluating FNA thyroid samples at the procedure time and to assess the concordance between FCM evaluations, paired conventional cytology, and final histology of removed thyroid gland. We also analyzed the integrity of nucleic acids after FCM evaluation to assure viability for molecular assessment. Our results demonstrated that FCM might improve timely and accurate patient management. Thyroid cytological examination, a key tool in preoperative thyroid nodule evaluation, is specific and accurate; some drawbacks are due to inadequate or indeterminate cytological reports and there is a need for an innovative approach overcoming the limits of traditional cytological diagnostics. Fluorescence laser confocal microscopes (FCM) is a new optical technique for allowing immediate digital imaging of fresh unfixed tissues and real-time assessment of sample adequacy and diagnostic evaluation for small biopsies and cytological samples. Currently, there are no data about the use of FCMs in the field of thyroid nodular pathology. The aims of this study were to test FCM technology for evaluating the adequacy of FNA samples at the time of the procedure and to assess the level of concordance between FCM cytological evaluations, paired conventional cytology, and final surgical histology. The secondary aim was to define the integrity of nucleic acids after FCM evaluation through NGS molecular analysis. Sample adequacy was correctly stated. Comparing FCM evaluation with the final histology, all cases resulting in malignant or suspicious for malignancy at FCM, were confirmed to be carcinomas (PPV 100%). In conclusion, we describe a successful application of FCM in thyroid preoperative cytological evaluation, with advantages in immediate adequacy assessment and diagnostic information, while preserving cellular specimens for permanent morphology and molecular analysis, thus improving timely and accurate patient management. [ABSTRACT FROM AUTHOR]

Embryological features were used for the first time as a taxonomic attribute to confirm the difference between closely related taxa in the subtribe Malaxidinae. It was shown that the branched shape of the suspensor in Liparis elegans and L. parviflora from the section Blepharoglossum distinguishes these species from the inflated suspensor without a neck, which is characteristic of the embryos of L. viridiflora and L. dendrochiloides from the section Cestichis. Differentiation of the development and shape of the suspensor is an additional embryological criterion in favor of separating the genus Blepharoglossum from the genus Liparis. [ABSTRACT FROM AUTHOR]

The biofilm formation by bacteria is a complex process that is strongly mediated by various genetic and environmental factors. Biofilms contribute to disease infestation, especially in chronic infections. It is, therefore important to understand the factors affecting biofilm formation. This study reports the role of a functional amyloid curli in biofilm formation at various abiotic surfaces, including medical devices, by an environmental isolate of Enterobacter cloacae (SBP-8) which has been known for its pathogenic potential. A knockout mutant of csgA, the gene encoding the major structural unit of curli, was created to study the effect of curli on biofilm formation by E. cloacae SBP-8. Our findings confirm the production of curli at 25 °C and 37 °C in the wild-type strain. We further investigated the role of curli in the attachment of E. cloacae SBP-8 to glass, enteral feeding tube, and foley latex catheter. Contrary to the previous studies reporting the curli production below 30 °C in the majority of biofilm-forming bacterial species, we observed its production in E. cloacae SBP-8 at 37 °C. The formation of more intense biofilm in wild-type strain on various surfaces compared to curli-deficient strain (ΔcsgA) at both 25 °C and 37 °C suggested a prominent role of curli in biofilm formation. Further, electron and confocal microscopy studies demonstrated the formation of diffused monolayers of microbial cells on the abiotic surfaces by ΔcsgA strain as compared to the thick biofilm by respective wild-type strain, indicating the involvement of curli in biofilm formation by E. cloacae SBP-8. Overall, our findings provide insight into biofilm formation mediated by curli in E. cloacae SBP-8. Further, we show that it can be expressed at a physiological temperature on all surfaces, thereby indicating the potential role of curli in pathogenesis. [ABSTRACT FROM AUTHOR]

Aim: Thyme has got good anti-inflammatory, antimicrobial, and antifungal properties and is reported to be very effective against oral microflora. The aim of the present study was to compare the antimicrobial activity of zinc oxide and thyme oil to zinc oxide and eugenol. Methods: Anti-bacterial efficacy of zinc oxide + eugenol and zinc oxide + thyme oil paste was evaluated against mature Enterococcus faecalis (E. faecalis) dentinal biofilm evaluated by the zone of inhibition (ZOI), biofilm disruption evaluated by colony-forming unit (CFU) reduction, and confocal laser scanning microscopy (CLSM). The red and green fluorescence thresholds were manually set according to their respective intensity and kept consistent for each sample. The ratios of live/dead cells were automatically calculated by the software. Results: The results of the present study showed that there was a statistically significant difference in antimicrobial efficacy for thyme oil compared to eugenol as evaluated by ZOI (P = 0.032). Thyme oil was significantly better than eugenol in antimicrobial efficacy according to the percentage of CFU reduction (P = 0.0422) in E. faecalis biofilm. Based on CLSM evaluation, there was no statistically significant difference (P = 0.172) between the two groups. Conclusion: Zinc oxide + Thyme oil paste, used in this study has shown promising antibacterial activity against E. faecalis and hence can be used as an alternative sealer.

Objective: To evaluate the feasibility of confocal laser microscopy (CLM) for intraoperative margin assessment as faster alternative to neurovascular structure‐adjacent frozen‐section examination (NeuroSAFE) during robot‐assisted radical prostatectomy (RARP). Patients and Methods: Surgical margins were assessed during 50 RARP procedures in patients scheduled for NeuroSAFE. Posterolateral sections were cut and imaged with CLM and further processed to conform with the NeuroSAFE protocol. Secondary resection (SR) was performed in case a positive surgical margin (PSM) was observed with NeuroSAFE. Afterwards, the CLM images were non‐blinded assessed for the presence of PSMs. The accuracy of both NeuroSAFE and CLM was compared with conventional histopathology. Agreement for detection of PSMs between NeuroSAFE and CLM was evaluated with Cohen's kappa coefficient. Procedure times were compared with a Wilcoxon signed‐ranks test. Results: In total, 96 posterolateral sections of RP specimens were evaluated for the presence of PSMs. CLM identified 15 (16%) PSMs and NeuroSAFE identified 14 (15%) PSMs. CLM had a calculated sensitivity, specificity, positive predictive value and negative predictive value of 86%, 96%, 80% and 98% respectively for the detection of PSMs compared to definite pathology. After SR, residual tumour was found in six of 13 cases (46%), which were all identified by both techniques. There was a substantial level of agreement between CLM and NeuroSAFE (κ = 0.80). The median procedure time for CLM was significantly shorter compared to NeuroSAFE (8 vs 50 min, P < 0.001). The main limitation of this study was the non‐blinded assessment of the CLM images. Conclusions: Compared to NeuroSAFE, CLM is a promising technique for intraoperative margin assessment and is able to reduce the time of intraoperative margin assessment. [ABSTRACT FROM AUTHOR]

Background: Reflectance confocal microscopy (RCM) has a defined in vivo morphology of vitiligo and re-pigmentation. Combination therapies seem more effective than monotherapies. Objective: We aim to describe the clinical and RCM features of re-pigmentation with combined narrowband ultraviolet B (NB-UVB) and piperine-based topical treatment in localized vitiligo. Methods: Eight patients enrolled at a single center received combined treatment: topical treatment was applied twice daily + NB-UVB twice weekly for 2 × 2-month periods. Clinical changes were analyzed by the Vitiligo Noticeability Scale (VNS) and percentage of re-pigmentation. The evaluator agreement was assessed. Predefined RCM features had the presence/absence of (i) blood vessels, (ii) dendritic cells, and the quantity of (i) an irregular honeycombed pattern and (ii) non-pigmented papillae. Clinical and RCM monitoring was performed at the baseline, 2, 3, 5, and 7 months. Results: Macules were “slightly less noticeable” with 25–50% re-pigmentation. Irregular honeycomb patterns and non-pigmented papillae were significantly less frequently observed, and in less extended areas (T1 vs. T2, p = 0.039; T0 vs. T1, p = 0.005 and T2 vs. T4, p = 0.033). Dendritic cells and blood vessels improved, with significant changes in blood vessels (T1 vs. T2, p = 0.005 and T3 vs. T4, p = 0.008). Conclusions: RCM confirms the morphological changes induced by combined treatment for localized vitiligo.

Aim: Thyme has got good anti-inflammatory, antimicrobial, and antifungal properties and is reported to be very effective against oral microflora. The aim of the present study was to compare the antimicrobial activity of zinc oxide and thyme oil to zinc oxide and eugenol. Methods: Anti-bacterial efficacy of zinc oxide + eugenol and zinc oxide + thyme oil paste was evaluated against mature Enterococcus faecalis (E. faecalis) dentinal biofilm evaluated by the zone of inhibition (ZOI), biofilm disruption evaluated by colony-forming unit (CFU) reduction, and confocal laser scanning microscopy (CLSM). The red and green fluorescence thresholds were manually set according to their respective intensity and kept consistent for each sample. The ratios of live/dead cells were automatically calculated by the software. Results: The results of the present study showed that there was a statistically significant difference in antimicrobial efficacy for thyme oil compared to eugenol as evaluated by ZOI (P = 0.032). Thyme oil was significantly better than eugenol in antimicrobial efficacy according to the percentage of CFU reduction (P = 0.0422) in E. faecalis biofilm. Based on CLSM evaluation, there was no statistically significant difference (P = 0.172) between the two groups. Conclusion: Zinc oxide + Thyme oil paste, used in this study has shown promising antibacterial activity against E. faecalis and hence can be used as an alternative sealer. [ABSTRACT FROM AUTHOR]

In search for natural products with antimicrobial properties for use in the prevention and treatment of peri-implantitis, the purpose of this investigation was to evaluate the antimicrobial activity of curcumin and xanthohumol, using an in vitro multi-species dynamic biofilm model including Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of curcumin (5 mM) and xanthohumol (100 μM) extracts, and the respective controls, were evaluated with 72-h biofilms formed over dental implants by their submersion for 60 seconds. The evaluation was assessed by quantitative polymerase chain reaction (qPCR), confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). For the data analysis, comparisons were tested applying ANOVA tests with post-hoc Bonferroni corrections to evaluate the antimicrobial activity of both extracts. With qPCR, statistically significant reductions in bacterial counts were observed for curcumin and xanthohumol, when compared to the negative control. The results with CLSM and SEM were consistent with those reported with qPCR. It was concluded that both curcumin and xanthohumol have demonstrated antimicrobial activity against the six bacterial species included in the dynamic in vitro biofilm model used. [ABSTRACT FROM AUTHOR]

The study assessed microhardness, microleakage of Class I, and the degree of conversion of composite resins preheated at 55 °C. This in vitro study evaluated two nanofilled composite resins (NCR), Filtek™ P60 and Filtek™ Z350 XT, used in the restorations. The degree of conversion (DC) was determined by Raman spectroscopy; the study compared the vibrational bands of the residual unpolymerized methacrylate C=C stretching band at 1640 cm−1 to the aromatic C–C stretching band at 1610 cm−1, used as an internal standard. Microleakage tests considered standard Class 1 cavities prepared on human premolars randomly divided. Preheating Filtek™ P60 and Filtek™ Z350 XT increased significantly VMH values (p = 0.0001). The highest microleakage occurred in preheated Filtek™ Z350 XT, while the lowest was in unheated Filtek™ P60. Unheated Filtek™ Z350 XT showed a significant increase of microleakage than unheated Filtek™ P60. (p = 0.024). Preheating of composite resins improved mechanical properties increasing DC and VMH. However, it also increases microleakage promoting bacterial penetration, secondary caries, marginal discoloration and hypersensitivity. [ABSTRACT FROM AUTHOR]

The copper(ii) complex [Cu(H2L1)2(H2O)2] with (2-hydroxyphenyl)phosphonic acid (H3L1) was synthesized. The composition and structure of the complex were determined by X-ray diffraction, elemental analysis, and IR spectroscopy. Quantum chemical calculations of the structure of the H3L1 molecule were performed by the density functional theory (DFT). The protonation constants of H3L1 and the stability constants of its complexes with Cu2+ in water were determined by potentiometric titration. The cytotoxic properties and the accumulation of the [Cu(H2L1)2(H2O)2] complex in human cervical adenocarcinoma HeLa cells were evaluated for the first time, thereby demonstrating that this complex is promising for further biological studies. [ABSTRACT FROM AUTHOR]

Abstract Background The native potatoes (Solanum tuberosum subsp. tuberosum L.) grown in Chile (Chiloé) represent a new, unexplored source of endophytes to find potential biological control agents for the prevention of bacterial diseases, like blackleg and soft rot, in potato crops. Result The objective of this study was the selection of endophytic actinobacteria from native potatoes for antagonistic activity against Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum, and their potential to suppress tissue maceration symptoms in potato tubers. This potential was determined through the quorum quenching activity using a Chromobacterium violaceaum ATCC 12472 Wild type (WT) bioassay and its colonization behavior of the potato plant root system (S. tuberosum) by means of the Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) targeting technique. The results showed that although Streptomyces sp. TP199 and Streptomyces sp. A2R31 were able to inhibit the growth of the pathogens, only the Streptomyces sp. TP199 isolate inhibited Pectobacterium sp. growth and diminished tissue maceration in tubers (p ≤ 0.05). Streptomyces sp. TP199 had metal-dependent acyl homoserine lactones (AHL) quorum quenching activity in vitro and was able to colonize the root endosphere 10 days after inoculation. Conclusions We concluded that native potatoes from southern Chile possess endophyte actinobacteria that are potential agents for the disease management of soft rot and blackleg.

The outer and internal cuticles in apple (Malus domestica Borkh.) leaves on the plain and in the mountains was studied using transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM). The outer cuticle consisted of lamellate and homogeneous layers of the cuticle proper and cuticular layer containing electron‐transparent plates and electron‐dense dendrites. Blue fluorescence predominated in the cell wall. The cuticle fluoresced green and red. The intensity of the red part of the spectrum in the cuticle increased with altitude, and the number of electron‐transparent plates increased within the cuticular layer. The cell wall on both leaf sides was the thinnest in the arid conditions (300 m). At an altitude of 600 m, under favorable temperature and rainfall conditions, the cuticle thickness increased due to the cuticular layer adjacent to the cell wall. The internal cuticle was distinguished by intense yellow or red autofluorescence, similar in color and spectrum to the outer cuticle. The outer and internal cuticles had cuticular folds. The average distance between the ridges of the internal cuticle was almost the same in the samples at different altitudes. The ridge height was maximum at 600 m. Research highlights: The altitudinal effects on outer and internal cuticles in apple leaves were studied.In the forest‐mountain zone, the cuticle thickness increased due to the cuticular layer adjacent to the cell wall.The cell wall was the thinnest in arid conditions. [ABSTRACT FROM AUTHOR]

The aim of the present in vitro study was to evaluate the intratubular penetration of three bioceramic sealers in root canal retreatment. Here, 30 single-rooted human teeth were instrumented with the Protaper Universal system and filled with gutta-percha and the epoxy-resin-based sealer AH Plus mixed with rhodamine B. After two weeks in a humid environment, they were re-instrumented with Reciproc Blue and divided into three groups according to the endodontic sealer to be used in the re-filling (n = 10): G1: CeraSeal, G2: TotalFill BC Sealer, G3: TotalFill BC Sealer HiFlow. For the filling, a single cone technique was used, and the respective sealers were mixed with fluorescein. The roots were then sectioned at 2, 5, and 8 mm (apical, medial, and coronal measurement points, respectively) from the apex, and the dentinal tubule penetration depth and percentage of penetration around the canal perimeter were evaluated by means of confocal laser scanning microscopy (CLSM). Penetration between groups was compared using the Kruskal−Wallis test, and within each group using the Wilcoxon test. Statistical significance was established at p < 0.05. A non-significant reduction was found in the penetration depths and in a percentage of penetration around the canal perimeter between AH Plus and the tested calcium-silicate-based sealers (p > 0.05). Consequently, this reduction may not affect the three-dimensional seal of the root canal system in a negative manner. The penetration depth and percentage of penetration around the canal perimeter at both the root canal treatment and retreatment were significantly reduced from the coronal to apical points in all groups (p < 0.05). [ABSTRACT FROM AUTHOR]

Abstract—The aim of the study was to investigate brain GABAergic neurons and synaptic terminals in early postnatal development and aging. Wistar rat brain slices at different stages of postnatal development (from postnatal day 7 to 24 months) were used. Immunohistochemical staining of GABA synthesizing enzyme (glutamate decarboxylase isoform 67 or GAD67) was performed to reveal GABAergic structures. We described morphological changes that occur in the GABAergic system during postnatal development. In particular, it has been shown that synaptic terminals are predominantly localized in cortical layer 1 in 7-day-old animals. Our findings suggest that Cajal–Retzius cells are not GABAergic. GABAergic neurons were found in young and old animals in the subventricular zone. We found that the epithelial layer of choroid plexus contains GAD67 by the end of the first month of postnatal development. In this regard, these epithelial cells may be a source of extrasynaptic GABA, which enters the cerebrospinal fluid and then nervous tissue of the brain. [ABSTRACT FROM AUTHOR]

Introduction. Currently, numerous studies are published by authors of different countries to demonstrate the effectiveness of noninvasive methods in the diagnosis of melanoma. Methods. A systematic search was conducted independently in the databases PubMed and Cochrane Central Register of Controlled Trials (CENTRAL) until April 2020 according to certain inclusion criteria. Data extraction was carried out independently, followed by generalization using descriptive tables. Due to the heterogeneity of the included studies and the impossibility of carrying out a meta-analysis in view of this, we performed a narrative description. Results. A total of 765 potential publications for inclusion were found and checked, of which 53 were included. By design, the studies were assigned to studies of simultaneous design – 40, to randomized clinical trials – 7, to meta-analysis – 6. Data in the included publications on 76802 skin neoplasms were reported, of which 9070 were melanomas. The extracted data were summarized in descriptive tables. Conclusion. With continuing technological progress, the development of noninvasive imaging technologies in the diagnosis of skin melanoma should follow the path of cost-effective, simple and accurate diagnosis.

The definitive diagnosis of patients with indeterminate biliary strictures remains challenging. Probe-based confocal laser endomicroscopy (pCLE) provides real-time histological assessment of bile duct tissues. Since no previous studies have evaluated the efficacy of pCLE under direct cholangioscopic visualization for biliary strictures that cannot be definitively diagnosed through endoscopic retrograde cholangiopancreatography using fluoroscopy, we prospectively assessed the feasibility and safety of this procedure in three cases. pCLE findings were obtained in three cases, providing accurate diagnoses. Additionally, no adverse event was reported. pCLE under direct cholangioscopic visualization for indeterminate biliary strictures might be feasible and safe, even though these strictures were not previously diagnosed using endoscopic retrograde cholangiopancreatography. Further studies with more cases are warranted to clarify the effectiveness of pCLE under direct cholangioscopic visualization., Competing Interests: None., (© 2024 The Author(s). DEN Open published by John Wiley & Sons Australia, Ltd on behalf of Japan Gastroenterological Endoscopy Society.)

Imprecision in breast conserving surgery results in high rates of take back to theatre for reexcision of margins. This paper reviews the various approaches to improving the precision of oncological margin control in breast conserving surgery. The review describes the rationale for improved tissue characterization over tumor localization and explores technology-free approaches, as well as progress being made to develop and test innovative technological solutions., (© 2024 The Author(s). Journal of Surgical Oncology published by Wiley Periodicals LLC.)

Background: The native potatoes (Solanum tuberosum subsp. tuberosum L.) grown in Chile (Chiloé) represent a new, unexplored source of endophytes to find potential biological control agents for the prevention of bacterial diseases, like blackleg and soft rot, in potato crops. Result : The objective of this study was the selection of endophytic actinobacteria from native potatoes for antagonistic activity against Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum, and their potential to suppress tissue maceration symptoms in potato tubers. This potential was determined through the quorum quenching activity using a Chromobacterium violaceaum ATCC 12472 Wild type (WT) bioassay and its colonization behavior of the potato plant root system (S. tuberosum) by means of the Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) targeting technique. The results showed that although Streptomyces sp. TP199 and Streptomyces sp. A2R31 were able to inhibit the growth of the pathogens, only the Streptomyces sp. TP199 isolate inhibited Pectobacterium sp. growth and diminished tissue maceration in tubers (p ≤ 0.05). Streptomyces sp. TP199 had metal-dependent acyl homoserine lactones (AHL) quorum quenching activity in vitro and was able to colonize the root endosphere 10 days after inoculation. Conclusions: We concluded that native potatoes from southern Chile possess endophyte actinobacteria that are potential agents for the disease management of soft rot and blackleg. [ABSTRACT FROM AUTHOR]

The morphologies of the internal organs of echinoderm larvae and juveniles are difficult to study using conventional optical microscopes because of their structural complexity and opaqueness. This paper describes an easy and rapid protocol involving Nile blue staining followed by benzyl alcohol/benzyl benzoate (BABB) clearing to overcome this limitation. This method was developed for a three‐dimensional (3D) analysis of the internal structures of advanced larvae and juveniles of echinoderms (the sea lily Metacrinus rotundus, the sea urchin Hemicentrotus pulcherrimus, and the sand dollar Scaphechinus mirabilis) and is suitable for obtaining serial optical images by confocal microscopy without the use of specific antibodies or special reagents for labeling. Nile blue is an easy‐to‐use stain that offers several advantages for confocal microscopy such as it can stain various tissues with strong fluorescent signals without substantial bleaching during observation. We found that the strong fluorescence signal of Nile blue quickly yielded clear high‐resolution optical section images for 3D reconstruction. BABB clearing rendered opaque larvae highly transparent. The clearing procedure was also easy and quick. During the process, agarose embedding prior to staining and clearing was found to be critical for handling the samples of less than 500‐μm length and stabilizing their orientations. To conclude, the protocol described is useful for performing a rapid and accurate 3D morphological analysis of echinoderm larvae and juveniles. [ABSTRACT FROM AUTHOR]

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This work was aimed at studying the structural organization of nucleolus and constitutive heterochromatin in different types of tanycytes during postnatal development and aging of rats. The distribution of nucleolus argentophilic proteins (nucleolin and nucleophosmin) and heterochromatin aggregates in tanycytes at various stages of postnatal development have been described for the first time using immunohistochemical methods and confocal laser microscopy. The heterogeneity of the size and number of nucleoli was demonstrated both in different tanycytes subpopulations and at different ages of an animal. This may indicate different levels of the tanycyte synthetic activity and the ability to proliferate during early postnatal development and aging. During aging, the distribution of heterochromatin aggregates varies among tanycyte subpopulations: α-tanycytes undergo intense heterochromatization, while β-tanycytes are characterized by a stable organization of the studied compartments of the cell nucleus. The data obtained significantly supplement the modern understanding of organization of the structure of the cell nucleus of tanycytes during normal development and aging. This can subsequently serve as a basis for establishing the role of these subnuclear structures in pathological processes. [ABSTRACT FROM AUTHOR]

The phase of the cell cycle can determine whether a cancer cell can respond to a given drug. We report here on the results of monitoring of real-time cell cycle dynamics of cancer cells throughout a live tumor intravitally using a fluorescence ubiquitination cell cycle indicator (FUCCI) before, during, and after chemotherapy. In nascent tumors in nude mice, approximately 30% of the cells in the center of the tumor are in G₀/G₁ and 70% in S/G₂/M. In contrast, approximately 90% of cancer cells in the center and 80% of total cells of an established tumor are in G₀/G₁ phase. Similarly, approximately 75% of cancer cells far from (> 100 µm) tumor blood vessels of an established tumor are in G₀/G₁. Longitudinal real-time imaging demonstrated that cytotoxic agents killed only proliferating cancer cells at the surface and, in contrast, had little effect on quiescent cancer cells, which are the vast majority of an established tumor. Moreover, resistant quiescent cancer cells restarted cycling after the cessation of chemotherapy. Our results suggest why most drugs currently in clinical use, which target cancer cells in S/G₂/M, are mostly ineffective on solid tumors. The results also suggest that drugs that target quiescent cancer cells are urgently needed.

Invasive cancer cells are a critical target in order to prevent metastasis. In the present report, we demonstrate real-time visualization of cell cycle kinetics of invading cancer cells in 3-dimensional (3D) Gelfoam® histoculture, which is in vivo-like. A fluorescence ubiquitination cell cycle indicator (FUCCI) whereby G0/G1 cells express a red fluorescent protein and S/G2/M cells express a green fluorescent protein was used to determine the cell cycle position of invading and non-invading cells. With FUCCI 3D confocal imaging, we observed that cancer cells in G0/G1 phase in Gelfoam® histoculture migrated more rapidly and further than cancer cells in S/G2/M phases. Cancer cells ceased migrating when they entered S/G2/M phases and restarted migrating after cell division when the cells re-entered G0/G1. Migrating cancer cells also were resistant to cytotoxic chemotherapy, since they were preponderantly in G0/G1, where cytotoxic chemotherapy is not effective. The results of the present report suggest that novel therapy targeting G0/G1 cancer cells should be developed to prevent metastasis.

Noha Khalifa Abo Aasy,1 Doaa Ragab,1,2 Marwa Ahmed Sallam,1,3 Doaa A Abdelmonsif,4,5 Rania G Aly,6 Kadria A Elkhodairy1 1Department of Industrial Pharmacy, Faculty of Pharmacy, Alexandria University, Alexandria 21521, Egypt; 2Department of Chemical and Biochemical Engineering, Faculty of Engineering, University of Western Ontario, London, Ontario, Canada; 3John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA; 4Department of Medical Biochemistry, Faculty of Medicine, Alexandria University, Alexandria, Egypt; 5Molecular Biology and Nanomedicine Labs, Centre of Excellence for Regenerative Medicine Research & Applications, University of Alexandria, Alexandria, Egypt; 6Department of Surgical Pathology, Faculty of Medicine, Alexandria University, Alexandria, EgyptCorrespondence: Noha Khalifa Abo AasyDepartment of Industrial Pharmacy, Faculty of Pharmacy, Alexandria University, 1-Azarita Square, Alexandria, EgyptTel +20 122 710 1058Fax +20 3 487 3273Email drnohakhalifa@yahoo.comIntroduction: This study was conducted to elucidate the chemopreventive potential, cytotoxic, and suppression of cellular metastatic activity of etodolac (ETD)-loaded nanocarriers.Methods: To esteem the effect of charge and composition of the nanovectors on their performance, four types of vectors namely, negative lipid nanovesicles; phosalosomes (N-Phsoms), positive phosalosomes (P-Phsoms), nanostructured lipid carriers (NLCs) and polymeric alginate polymer (AlgNPs) were prepared and compared. ETD was used as a model cyclo-oxygenase-2 (COX-2) inhibitor to evaluate the potency of these nanovectors to increase ETD permeation and retention through human skin and cytotoxicity against squamous cell carcinoma cell line (SCC). Moreover, the chemopreventive activity of ETD nanovector on mice skin cancer model was evaluated.Results: Among the utilized nanovectors, ETD-loaded N-Phsoms depicted spherical vesicles with the smallest particle size (202.96±2.37 nm) and a high zeta potential of −24.8±4.16 mV. N-Phsoms exhibited 1.5, and 3.6 folds increase in the ETD amount deposited in stratum corneum, epidermis and dermis. Moreover, cytotoxicity studies revealed a significant cytotoxic potential of such nanovector with IC50=181.76 compared to free ETD (IC50=982.75), correlated to enhanced cellular internalization. Its efficacy extended to a reduction in the relative tumor weight with 1.70 and 1.51-fold compared to positive control and free ETD, that manifested by a 1.72-fold reduction in both COX-2 and proliferating cell nuclear antigen mRNA (PCNA-mRNA) levels and 2.63-fold elevation in caspase-3 level in skin tumors relative to the positive control group with no hepato-and nephrotoxicity.Conclusion: Encapsulation of ETD in nanovector enhances its in-vitro and in-vivo anti-tumor activity and opens the door for encapsulation of more relevant drugs.Keywords: COX-2, confocal laser microscopy, skin cancer, metastasis, cytotoxicity, chemoprevention

The aim of the present in vitro study was to evaluate the intratubular penetration of three bioceramic sealers in root canal retreatment. Here, 30 single-rooted human teeth were instrumented with the Protaper Universal system and filled with gutta-percha and the epoxy-resin-based sealer AH Plus mixed with rhodamine B. After two weeks in a humid environment, they were re-instrumented with Reciproc Blue and divided into three groups according to the endodontic sealer to be used in the re-filling (n = 10): G1: CeraSeal, G2: TotalFill BC Sealer, G3: TotalFill BC Sealer HiFlow. For the filling, a single cone technique was used, and the respective sealers were mixed with fluorescein. The roots were then sectioned at 2, 5, and 8 mm (apical, medial, and coronal measurement points, respectively) from the apex, and the dentinal tubule penetration depth and percentage of penetration around the canal perimeter were evaluated by means of confocal laser scanning microscopy (CLSM). Penetration between groups was compared using the Kruskal−Wallis test, and within each group using the Wilcoxon test. Statistical significance was established at p < 0.05. A non-significant reduction was found in the penetration depths and in a percentage of penetration around the canal perimeter between AH Plus and the tested calcium-silicate-based sealers (p > 0.05). Consequently, this reduction may not affect the three-dimensional seal of the root canal system in a negative manner. The penetration depth and percentage of penetration around the canal perimeter at both the root canal treatment and retreatment were significantly reduced from the coronal to apical points in all groups (p < 0.05).

High-definition endoscopy is one essential step in the initial diagnosis of inflammatory bowel disease (IBD) characterizing the extent and severity of inflammation, as well as discriminating ulcerative colitis (UC) from Crohn's disease (CD). Following general recommendations and national guidelines, individual risk stratification should define the appropriate surveillance strategy, biopsy protocol and frequency of endoscopies. Beside high-definition videoendoscopy the application of dyes applied via a spraying catheter is of additional diagnostic value with a higher detection rate of intraepithelial neoplasia (IEN). Virtual chromoendoscopy techniques (NBI, FICE, I-scan, BLI) should not be recommended as a single surveillance strategy in IBD, although newer data suggest a higher comparability to dye-based chromoendoscopy than previously assumed. First results of oral methylene blue formulation are promising for improving the acceptance rate of classical chromoendoscopy. Confocal laser endomicroscopy (CLE) is still an experimental but highly innovative endoscopic procedure with the potential to contribute to the detection of dysplastic lesions. Molecular endoscopy in IBD has taken application of CLE to a higher level and allows topical application of labeled probes, mainly antibodies, against specific target structures expressed in the tissue to predict response or failure to biological therapies. First pre-clinical and in vivo data from label-free multiphoton microscopy (MPM) are now available to characterize mucosal and submucosal inflammation on endoscopy in more detail. These new techniques now have opened the door to individualized and highly specific molecular imaging in IBD in the future and pave the path to personalized medicine approaches. The quality of evidence was stated according to the Oxford Center of evidence-based medicine (March 2009). For this review a Medline search up to January 2021 was performed using the words “inflammatory bowel disease,” “ulcerative colitis,” “crohn's disease,” “chromoendoscopy,” “high-definition endoscopy,” “confocal laser endomicroscopy,” “confocal laser microscopy,” “molecular imaging,” “multiphoton microscopy.”

The aim of the study was to investigate the organization of GABAergic axosomatic synaptic terminals in the parietal, cingular and piriform cortical areas of the rat brain. The methodological approach of the study was an immunohistochemical staining for glutamate decarboxylase – the GABA-synthetic enzyme. Samples were analyzed using confocal laser microscopy. Brain of adult male Wistar rats (n = 11) were studied. As a result of the work, information of the morphological organization of pre-synaptic terminals from distinct cortical areas was obtained. It has been found that sizes of GABAergic synapses and their distribution densities onto a pyramidal soma in distinct cortical regions are significantly different (P < 0.05). It has been shown that confocal laser microscopy can provide objective information for investigating synapse structure. The obtained results allow a more complete assessment of inhibitory structures of the brain and their interaction with glutamatergic pyramidal neurons. The methodological approach used in the study can help to identify morphological signs of GABAergic system dysfunction. [ABSTRACT FROM AUTHOR]

According to the obtained data over the last decades, astrocytes play a crucial role in neuronal homeostasis. Therefore, the software for assessing morphological changes in astrocytes is not sufficiently covered in the literature. Hence, this article examines a two- and three-dimensional quantitative approach to astrocyte structure analysis based on freely available software. The brains of mature male Wistar rats were used (n = 5). Double immunocytochemical GFAP/GLT-1 staining was performed to obtain fluorescent confocal images. The article proposes the implementation of recent analysis approaches using freely available software to obtain qualitive and quantitative information about the structural features of heterogeneous cells of the central nervous system such as astrocytes. Moreover, the proposed approach provides standardization of data that necessary for comparing the results obtained by different scientific groups. [ABSTRACT FROM AUTHOR]

Background.This study compared the effect of smear layer on the penetration depth and push-out bond strength of various root canal sealers. Methods.A total of 90 extracted human mandibular premolars were assigned into 2 groups: smear layer preserved and smear layer removed. Then the roots were further divided into 3 subgroups according to the sealer tested: AH 26, BioRoot RCS and MTA Plus. Obturation was performed with gutta-percha and the relevant sealer was mixed with 0.1% rhodamine B. Three 1-mm-thick slices were obtained from the mid-third area of each root. Two slices were selected for the push-out test and the remaining slice was used to calculate the dentinal tubule penetration depth and percentage. Results.The retention of MTA Plus and BioRoot RCS was higher than that of AH 26 when the smear layer was preserved (P

The development of drug delivery systems using nanocarriers requires intraorgan imaging techniques for evaluating the distribution of nanocarriers. In this study, we evaluated the tissue-clearing techniques for the imaging of polymeric nanoparticles, a nanocarrier, in the liver used as a model of pigment-rich organ in mice. The intraorgan imaging method of polymeric nanoparticles was examined without sectioning of organ samples for evaluating the delivery efficiency in preclinical studies. DiI-loaded polymeric nanoparticles and fluorescence-tagged tomato lectin for fluorescence labeling of liver general structures were intravenously administered to mice. Tissue-clearing treatment of the mouse liver was performed using ClearT2, ScaleSQ(0), clearing agent comprising fructose, urea, and glycerol for imaging (FUnGI), clear unobstructed brain/body imaging cocktails and computational analysis (CUBIC), and modified CUBIC techniques. Intraorgan fluorescence imaging in the liver was performed by confocal laser microscopy. ClearT2 treatment exhibited insufficient clearing capability in the mouse liver. Although CUBIC treatment exhibited the best clearing capability, the CUBIC caused DiI leakage. ScaleSQ(0), FUnGI, and modified CUBIC treatments exhibited better clearing capability than ClearT2 technique while preserving the DiI. In the fluorescence imaging, the CUBIC and modified CUBIC exhibited deeper visualization than with the ScaleSQ(0) and FUnGI; however, the CUBIC led to a change in DiI distribution. The modified CUBIC enabled the deepest visualization while preserving the distribution of DiI. The intraorgan imaging method was established using modified CUBIC technique by the intravenous administration of fluorescence-tagged tomato lectin for evaluating the distribution of polymeric nanoparticles in mouse pigment-rich organs. [ABSTRACT FROM AUTHOR]

Gastric cancer is a malignant tumor with a very high fatality rate, which is extremely harmful and disturbing. Gastric intraepithelial neoplasia (GIN) is a symptom of gastric cancer. If not treated in time, it will turn into early gastric cancer. Confocal laser microscopy is the most advanced microscope technology, which is of great significance for the diagnosis and observation of early gastric cancer and gastric intraepithelial neoplasia. The purpose of this article is to study the specific effect of confocal laser microendoscopy on the clinical diagnosis of early gastric cancer and gastric intraepithelial neoplasia. From 2017.6-2019.8, patients with gastritis, gastric intraepithelial neoplasia and a total of 20 early-stage gastric cancer patients have the image of the research pair. Under the condition of obtaining the patient's consent, all patients will be subjected to general microendoscopy. After the patient's status is adjusted, the confocal laser microendoscopy will be performed, and the study will be conducted according to the relevant diagnostic criteria. The results of the study showed that the accuracy of confocal laser microscopy in the diagnosis of early gastric cancer was 96.3%, the sensitivity was 92.5%, and the specificity was 97.6%. The accuracy of distinguishing gastric cancer lesions from non-cancerous lesions was 95.8%. The detection accuracy of intraepithelial neoplasia was 94.9%. It can be seen that confocal laser microscopy can effectively distinguish gastric cancer from non-cancerous lesions, thereby accurately diagnosing early gastric cancer and gastric intraepithelial neoplasia. [ABSTRACT FROM AUTHOR]

Purpose: To assess the effects of a recently developed ultrasonic scaler tip, composed mainly of copper,on the surfaces of several dental materials. Materials and Methods: Representative samples of dental materials, including titanium, type II gold, cobalt-chromium alloy, zirconia, and porcelain, were prepared.Three ultrasonic scaler tips of conventional metal (CM), carbon composite (CC), and copper alloy (CA) were prepared. To simulate ultrasonic scaling in an oral environment, 30 g of force was applied using a doublepan balance, and the scaler tip was allowed to move horizontally 5 mm for three consecutive cycles of 20seconds each. The power of the scaler tip was set to intermediate according to the manufacturer's advice.The surface morphology of each dental material was examined using scanning electron microscopy and confocal laser scanning microscopy. Statistical analysis was performed through one-way analysis of variance and post hoc Scheffé test. All values were considered significant when P < .05. Results: Surface alterations of titanium, type II gold, and cobalt-chromium alloy by the CM tip were much greater than those caused by the CC and CA tips. No alterations were created on the zirconia surface by the CM, CC, or CA tips. On the porcelain surface, surface roughness (Ra) induced by the CM tip was 1.86 and 1.72 times higher than that produced by the CC and CA tips, respectively (P < .001). Conclusions: Within the limitations of this study,the surface alterations induced by CC and CA tips on the surfaces of dental materials were comparable.Therefore, this novel ultrasonic copper alloy scaler tip may possibly be used for the maintenance of implant prostheses. Int J Oral Maxillofac Implants 2012;27:801-810. [ABSTRACT FROM AUTHOR]

The role of transcription factors in the pathogenesis of pituitary adenomas is extremely controversial.The aim of the study was to investigate the role of the transcription factor Neuro D1 in various types of pituitary adenomas.Materials and methods. A comparative clinico-morphological study was carried out with immunohistochemical analysis and confocal microscopy of the expression of the transcription factor NeuroD1, six adenohypophysis hormones and Ki-67 in 40 pituitary adenomas and 9 normal pituitary glands.Results. NeuroD1 was expressed in all cases and types of adenomas. The expression level of the transcription factor in adenomas was significantly different from that in the normal pituitary gland (p = 0.006). The average number of cells with expression of NeuroD1 in all tumors was higher than in the normal pituitary gland.Conclusion. NeuroD1 plays one of the key roles in the pathogenesis of pituitary adenomas, regardless of their hormonal status.