搜索结果：Microscopy research and technique

Diabetes mellitus (DM) induces endocrine and exocrine dysfunction, including salivary gland impairment. While the sublingual gland (SLG) contributes to saliva production, its involvement in diabetes remains understudied. Agmatine (AGM), a naturally occurring polyamine, exhibits antioxidant and anti-inflammatory properties; however, its protective effects on the SLG and pancreas in diabetes have remained largely uninvestigated. This study introduces new insights into SLG injury in diabetes and the dose-dependent protective benefits of AGM through multi-level analysis. Male Wistar rats were rendered diabetic by a single intraperitoneal STZ injection (65 mg/kg) and treated with either low-dose (40 mg/kg) or high-dose (100 mg/kg) AGM for two weeks. SLGs and the pancreas were analyzed by histological, immunohistochemical, and biochemical methods. Mucin secretion (MUC-5B) was measured, oxidative stress markers and pro-inflammatory cytokines were quantified, and structural changes were evaluated using H&E, Alcian Blue/PAS, and Mallory's trichrome staining. Immunohistochemistry included α-SMA and CK17 for SLG and insulin receptor for the pancreas. STZ-induced diabetes resulted in a substantial reduction in MUC-5B, increased oxidative stress and pro-inflammatory markers, damage to acinar and myoepithelial cells, and injury to pancreatic β-cells. Agmatine administration enhanced these parameters in a dose-dependent manner, with high-dose treatment reinstating mucin secretion, regulating oxidative and inflammatory indicators, and maintaining histological and immunohistochemical integrity in both the sublingual gland and pancreas. AGM confers dose-dependent protection against SLG dysfunction and pancreatic damage in diabetic rats via antioxidant, anti-inflammatory, and cytoprotective mechanisms. These findings highlight the therapeutic potential of AGM in preventing salivary gland dysfunction and maintaining pancreatic function in diabetes.

BACKGROUND: Accurate malaria diagnosis is critical not only for prescribing correct treatment but also for ensuring effective case management, preventing drug resistance, and interrupting transmission. Despite this importance, laboratory diagnostic error driven by multifactorial causes yielding both false-positive and false-negative results remains a persistent yet underrecognized challenge, particularly in endemic regions. This study aimed to compare routine malaria microscopy with Loop-Mediated Isothermal Amplification (LAMP) to determine the prevalence of malaria misdiagnosis by routine microscopy in Northwest Ethiopia. METHODS: A health facility&#x2013;based cross-sectional study was conducted at four selected health centers from July 1 to August 30, 2024. A convenience sampling technique was applied to recruit study participants. A total of 384 malaria-suspected individuals whose routine microscopy results recorded at health centers were re-evaluated using a LAMP molecular assay results performed on venous blood samples. The data were entered into Microsoft Office Excel and exported to SPSS version 27 for analysis. Descriptive statistics were used to estimate malaria misdiagnosis by routine microscopy, and the McNemar&#x2019;s test was applied to assess statistical significance, with a p-value&#x2009;<&#x2009;0.05 considered statistically significant. RESULTS: Among the 384 study participants (116 microscopy-positive and 268 microscopy-negative), 30.0% of routine malaria microscopy results were discordant when compared with LAMP findings. The discordant results comprised false-positive results (7.6%), false-negative results (18.8%), and species misidentification (3.6%). McNemar&#x2019;s test demonstrated a statistically significant difference between microscopy and LAMP results (p&#x2009;<&#x2009;0.001). CONCLUSIONS AND RECOMMENDATIONS: The current study showed a high prevalence of malaria misdiagnosis; false negatives, false positives, and species misidentification by routine microscopy. Although this undermines malaria control efforts, microscopy remains the most accessible and WHO-endorsed diagnostic method in resource-limited settings. Strengthening its accuracy through continuous training, quality assurance, and supervision is therefore essential. While molecular methods such as LAMP offer superior sensitivity and specificity, microscopy continues to serve as the operational gold standard due to its practicality and affordability. CLINICAL TRIAL NUMBER: Not applicable.

Cellular imaging is important in understanding drug pharmacokinetics and dynamics. As such, it is crucial that the drug is unmodified when performing these studies, to neither inhibit nor change its action and properties. Historically, fluorescence microscopy has been used for drug imaging due to its high sensitivity and ease of use, but bulky fluorescent tags have the potential to cause off-target effects and result in a change in the pharmacokinetic properties. The use and development of small optical tags are therefore attractive, as cellular systems can be probed with minimal perturbation to the cellular environment and the native kinetics of a drug. Bio-orthogonal Raman imaging makes use of molecular vibrations that are seldom observed in nature to determine spatial localization. Spontaneous Raman scattering can be used to achieve minimally labeled drug localization but is relatively slow and has a low spatial resolution when compared to fluorescence microscopy. Faster image acquisition and higher spatial resolution can be achieved by using stimulated Raman scattering (SRS), a powerful technique that is often used for native cellular imaging. The use of either intrinsically bio-orthogonal drugs or those with a small tag added allows Raman scattering to be used as a companion imaging diagnostic tool. This work assesses the localization of a covalently binding inhibitor of Bruton's tyrosine kinase, ibrutinib, using fluorescently labeled and bio-orthogonally Raman labeled analogues as companion diagnostic tools. Localization of these analogues was determined using fluorescence and Raman microscopies, and inhibitor retention was proportional to the expression of the kinase. Significant retention of the fluorescent analogue was observed independent of kinase expression, indicating significant nonspecific binding. Drug-induced effects were also explored using spectral phasor analysis of hyperspectral SRS data to assess lipid metabolism, where BTK inhibition was shown to cause an increase in the lipid content and change in the lipid type, which was proportional to kinase expression. This work showcases the advantages of Raman scattering techniques over fluorescence as companion imaging diagnostic tool and as a method of assessing phenotypic lipid shifts upon treatment with an anticancer drug.

In the present study, the adhesive organ (AO) of Hara hara, a hillstream catfish, has been investigated using scanning electron microscopy, histology, and glycoprotein, protein, and lectin histochemistry. The AO is a disc-like structure present on the ventral side of the body. It is distinguished into a major flat central region and a narrow flap-like region at the periphery of the central region. The free surface of the AO is characteristically differentiated into plaques separated by furrows. Surface epithelial cells in the plaques are modified into unicellular spine-like unculi. The furrow epithelium contains three types of unicellular gland cells (the mucous goblet cells, the club cells, and the sacciform cells), along with sensory structures such as the taste buds (TBs). Gland cells are involved in the elaboration of various glycoproteins (oxidizable vicinal diols, carboxyl groups, and O-sulphate esters); protein moieties (basic proteins, cysteine-bound sulphydryl groups, keratin, tyrosine, protein bound-NH2 groups, and elastin); and glycans (galactose, N-Acetylgalactosamine, and fucose). Presence of these glycoprotein and protein moieties elaborated by the cellular component of the AO has been associated to play an important role in maintaining structural integrity of the epithelium, keratinization, protection against pathogens, and adhesion of the fish to the substratum. The TBs located at the summit of the tubercles in the furrow epithelium may serve as an adaptation to enhance sensitivity to food selection and environmental perception. Overall, this study provides a comprehensive understanding of the structural and functional adaptations that enable H. hara to maintain effective attachment and thrive in turbulent hillstream environments.

BACKGROUND: The restoration of endodontically treated premolars has been a controversial topic. Whether a post or a more conservative option should be used remains inconclusive. The present in vitro study aimed to evaluate the fracture strength and failure pattern of different partial coverage restoration approaches for endodontically treated upper premolars in comparison to the post, core and crown approach. METHODS: Thirty-five maxillary premolars were endodontically treated and assigned to five groups (n&#x2009;=&#x2009;7): endocrown (E), endocrown with buccal veneer (EB), overlay (O), overlay with buccal veneer (OB), and post, core, and crown (P). All restorations were fabricated from lithium disilicate ceramics and subjected to vertical static loading until failure. Fracture strength was recorded, and failure patterns were examined using a stereomicroscope and scanning electron microscopy. Data were analyzed with Welch one-way ANOVA and Games&#x2013;Howell post hoc test (&#x3b1;&#x2009;=&#x2009;0.05). RESULTS : Significant differences were found among groups (p&#x2009;<&#x2009;0.001) with a large effect size (partial eta squared&#x2009;=&#x2009;0.83 (95% CI; 0.59 to 0.91). The highest mean fracture strength was observed in group P (1676.29&#x2009;&#xb1;&#x2009;191.25&#xa0;N), followed by OB (1475.00&#x2009;&#xb1;&#x2009;198.72&#xa0;N), E (1374.71&#x2009;&#xb1;&#x2009;371.91&#xa0;N), EB (1047.41&#x2009;&#xb1;&#x2009;163.90&#xa0;N), and O (987.99&#x2009;&#xb1;&#x2009;125.01&#xa0;N). Groups P and OB demonstrated significantly higher strength than EB and O. Group P exhibited only repairable restoration fractures, whereas the other groups predominantly showed catastrophic failures. CONCLUSIONS: Within the limitations of this in vitro study, post, core, and crown restorations provided superior fracture strength and the most favorable failure patterns under static axial loading. While overlays with buccal veneers demonstrated promising mechanical resistance thresholds, their clinical application should be considered with caution. These designs may serve as a potential conservative alternative, although further research involving dynamic fatigue and oblique loading is required to confirm their long-term clinical durability in the complex oral environment. CLINICAL RELEVANCE: The findings of this in-vitro study provide insight into the fracture strength and failure tendencies of different restorative approaches for endodontically treated maxillary premolars. While post, core, and crown restorations demonstrated high load-bearing capacity and predominantly favourable failure patterns, overlay designs with a buccal veneer showed comparable strength but a greater tendency toward catastrophic fractures. These observations highlight a potential balance between structural preservation and failure reparability. Given that static axial loading was used, direct clinical interpretation should be approached cautiously. Treatment planning should integrate these biomechanical considerations with patient-specific factors, functional demands, and clinical judgment when selecting restorative approaches.

To compare unsedated noncontact specular microscopy imaging techniques for the canine corneal endothelium and identify the most effective technique. Nineteen eyes of 10 systemically healthy, staff-owned dogs with clinically normal corneas were studied. Six imaging groups were divided according to the different techniques used in all eyes: central focusing, with (CP) or without (CN) induction of an indirect pupillary light reflex (PLR); peripheral focusing with (PP) or without (PN) an indirect PLR; PP with application of a contact lens (CL); and gaze fixation with a treat (GF). The success rates, imaging times, and endothelial cell indices were compared. Success rates differed significantly among the techniques, with PP and CL achieving significantly higher success rates than CN. When only successful images were analyzed, the mean imaging time showed no statistically significant differences between the groups. In the sensitivity analysis, PP required significantly shorter imaging times than CN and PN. The Endothelial cell indices (cell count, endothelial density, and hexagonality) did not differ significantly among the techniques. Peripheral focusing combined with the induction of an indirect PLR (PP) achieved significantly higher success rates and shorter imaging times than other techniques, suggesting that this technique provides the most reliable and efficient approach for evaluating the canine corneal endothelium with noncontact specular microscopy under unsedated conditions.

Loiasis is a filarial disease caused by Loa loa, endemic to Central and West Africa. Cases are observed in migrants and occasionally travellers. Its diagnosis may be difficult due to the unspecific clinical picture and the high percentage of people who do not have microscopically detectable circulating microfilariae. We performed a landscape analysis of the laboratory-based diagnostic techniques available for loiasis and of their estimated sensitivity and specificity. We performed a systematic review of cross-sectional, cohort, case-control, diagnostic accuracy, and clinical trial studies published in PubMed, EMBASE, and CENTRAL (searched on March 26th 2025), applying diagnostic assays for human loiasis. When possible, a proportional meta-analysis was performed by estimating sensitivity and specificity separately against eligible reference tests (direct assays or presence of "eyeworm" or their composite or latent class analysis) for each technique category (microscopy of thick smears or concentrated blood, PCR-based or LAMP-based molecular assay, and ELISA-based or rapid -RDT- serological assays). A random-effects model with the DerSimonian-Laird approach was applied. Study quality was evaluated using the Newcastle-Ottawa Scale. Ninety-nine publications were included in the landscape analysis and 27 were also eligible for performance assessment. Microscopy was applied in 91/99 (91.9%) studies, PCR-based techniques in 29/99 (29.3%), LAMP in 4/99 (4.0%), and serological assays in 19/99 (19.2%). Within techniques categories, characteristics were highly heterogeneous. Sensitivities ranged from 75.0-98.3% for thick blood smears, 48.2-99.9% for blood concentration techniques, 80.6-98.4% for PCR-based techniques, 88.5-98.1% for LAMP-based techniques, 81.9-90.5% for ELISA seroassays, and 43.6-88.0% for RDTs. Cross-reactivity of L. loa-specific seroassays with M. perstans and other parasitoses was limited (<10%). Assays for the diagnosis of loiasis are not standardized. ELISA-based serology and possibly PCR-based methods may be appropriate for screening. Microscopy must be performed even in case of negativity on screening when epidemiological or clinical factors suggestive of loiasis are present to plan safe treatment.

The pink bollworm, Pectinophora gossypiella Saunders is a major pest of cotton, notorious for its high reproductive potential and rapid evolution of resistance to Bacillus thuringiensis (Bt) toxins. Despite its economic significance, detailed knowledge of its reproductive anatomy and egg ultrastructure has remained limited, constraining the development of advanced molecular control strategies such as CRISPR/Cas9-based genome editing. The present study provides the first comprehensive characterization of the reproductive system and egg surface morphology of P. gossypiella using stereomicroscopy and scanning electron microscopy (SEM) techniques. The male reproductive system consists of fused, bean-shaped testes, seminal vesicles, duplex and simplex ejaculatory ducts, and paired accessory glands. The female reproductive system comprises paired ovaries with four polytrophic ovarioles per ovary, lateral and common oviducts, accessory glands, corpus bursae, and spermathecal glands. Eggs are oval, dorsoventrally flattened, exhibit a reticulated chorion with distinct micropylar and aeropylar regions. SEM images revealed 6-9 rosette cells encircling a circular micropylar plate, 14-19 first order and 17-23&#x2009;s order ribs, and 250-291 polygonal surface cells. The structural features of P. gossypiella eggs reveal key sites for sperm entry, aeropylar respiration, and candidate zones for microinjection in gene editing applications. These findings establish a morphological baseline critical for optimizing embryo manipulation and ribonucleoprotein (RNP) delivery in lepidopteran genome editing. This study represents a pioneering effort to integrate classical egg morphology with molecular entomology, thereby advancing precision genetic interventions aimed at resistance management and population suppression in P. gossypiella.

Immunohistochemistry is a tissue-based technique that enables in situ visualization of specific molecules, most commonly proteins or peptides, within preserved cellular and tissue architecture through antigen-antibody specificity. By using labeled antibodies, immunohistochemistry allows spatially resolved and cell type-specific detection of target molecules and maintains the morphological context, thereby providing both localization and relative expression information at the light or electron microscopy level. Owing to this integrative capacity, immunohistochemistry has become an essential tool in biological research and diagnostic pathology for characterizing molecular distribution and tissue organization across animal and plant systems. The detection techniques based on fluorescence and enzymes (chromogenic) are compared in terms of their repeatability, multiplexing capability, sensitivity, and specificity. With a focus on protocol standardization and methodological rigor, important procedural processes are covered, including fixation, antigen extraction, antibody optimization, signal detection, and quality control. Additionally, the immunohistochemistry technique remains essential for locating and identifying immunogenic substances in a variety of biological settings. Therefore, drawing on more than two decades of laboratory experience, this narrative review summarizes the history of the immunohistochemistry assay, its underlying principles and procedures, common immunohistochemistry protocol, methodological process, essential reagents, infrastructure, and best-practice principles required for the establishment and optimization of a high-quality immunohistochemistry laboratory.

Andrographolide (AG), a natural diterpenoid compound derived from Andrographis paniculata, exhibits potential against non-alcoholic steatohepatitis (NASH). However, its therapeutic utility is limited by poor solubility, short half-life, and low bioavailability. This study aimed to enhance AG's efficacy by encapsulating it into exosome-like nanoparticles (ELNs) isolated from Nauclea officinalis (N. officinalis, Rubiaceae family) and evaluating its anti-NASH activity in WRL68 cells. This study isolated Nauclea officinalis-derived ELNs (N-ELNs) via ultracentrifugation. Loganin, andrographolide, and asiatic acid were detected in N-ELNs using the HPLC method. RNA sequencing and target gene analysis identified miRNAs in N-ELNs and their cross-kingdom targets in the human genome. AG-loaded N-ELNs (AG-N-ELNs) were prepared using a passive loading method and characterized by transmission electron microscopy, nanoparticle tracking analyzer, and high-performance liquid chromatography. Their effects were tested in free fatty acid (FFA)-exposed WRL68 cells. As for the results, AG suppressed Nuclear Factor kappa-B p50 (NF-&#x3ba;B p50) activation, downregulated NLRP3 expression, and reduced pro-inflammatory cytokines (IL-1&#x3b2;, IL-6, TNF-&#x3b1;) in WRL68 cells. It also restored mitochondrial membrane potential and decreased reactive oxygen species (ROS). N-ELNs targeted the ACC/ CPT1 axis to alleviate lipid deposition, reducing total cholesterol and triglyceride levels. AG-N-ELNs outperformed both AG and N-ELNs individually, demonstrating superior anti-NASH activity through synergistic effects of AG's anti-inflammatory/antioxidant properties and N-ELNs' miRNA-mediated metabolic regulation. In conclusion, AG-N-ELNs effectively alleviate NASH-like pathological features in vitro through multi-target regulation, offering a promising strategy for NASH treatment. Further in vivo validation and formulation optimization are warranted.

The global rise of extensively drug-resistant Acinetobacter baumannii, particularly biofilm-forming strains, has drastically limited treatment options and created an urgent need for novel therapies to restore antibiotic efficacy. This study explored p-coumaric acid (p-CA) as a potential dual-action agent to combat biofilm-associated XDR Acinetobacter baumannii infections and restore imipenem efficacy. Among 100 clinical Acinetobacter baumannii isolates, 32 were identified as XDR and exhibited resistance to imipenem. The antimicrobial and antibiofilm efficacy of p-CA was systematically evaluated through comprehensive in vitro assays and an in vivo rat infection model. Minimum inhibitory concentrations were determined via the broth microdilution method, and the potential modulation effect on imipenem efficacy was investigated. To assess biofilm inhibition and disruption, quantitative analyses were performed using the crystal violet staining technique, complemented by evaluating its impact on the bacterial cell surface hydrophobicity and exopolysaccharide production. Biofilm structural changes were analyzed via light, scanning electron, and confocal laser scanning microscopy. Additionally, the expression levels of key biofilm-associated genes were quantified via quantitative reverse transcription PCR. The p-CA exhibited potent antimicrobial activity against the tested isolates (MIC: 512&#xa0;&#xb5;g/mL) and synergized with imipenem, reducing its MIC by 512-fold. At subinhibitory concentrations (&#xbc;-&#xbd; MIC), it inhibited biofilm formation (66.2-80.5%, p&#x2009;<&#x2009;0.05) and disrupted pre-formed biofilms (45.8-71.3%, p&#x2009;<&#x2009;0.05), likely via altered cell surface hydrophobicity and reduced EPS production. Microscopic imaging corroborated these findings, revealing substantial structural degradation of biofilms upon treatment. At the molecular level, p-CA significantly downregulated (p&#x2009;<&#x2009;0.05) the key biofilm-associated genes (abaI, bfmR, bap, csuE, and pgaB), as quantified by RT-qPCR. In vivo, the p-CA/imipenem combination significantly enhanced the survival rates (100%, p&#x2009;<&#x2009;0.05) and reduced the lung bacterial burden (p&#x2009;<&#x2009;0.001). Histopathological examination showed near-complete restoration of alveolar architecture by 72&#xa0;h post-treatment in the combination therapy group. These findings position p-CA as a promising dual-action adjuvant against XDR Acinetobacer baumannii infections, particularly in biofilm-associated contexts. It combines direct antimicrobial activity, biofilm disruption, and synergy with imipenem to address critical treatment gaps.

This study aimed to develop polylactic-co-glycolic acid (PLGA) microspheres encapsulating dezocine and evaluate their efficacy in managing postoperative pain and mitigating cognitive impairment. Dezocine-loaded PLGA microspheres (DEZ@PLGA MS) were fabricated using a single-emulsion solvent evaporation technique and were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and in vitro release studies. A rat model of open reduction and internal fixation for right hindlimb tibial plateau fracture was established in healthy male rats. Following modeling, rats were randomly divided into four groups (n&#x2009;=&#x2009;6 per group) and received intramuscular injections of equal volumes of normal saline, blank PLGA microsphere suspension, dezocine injection (5&#xa0;mg/kg), or DEZ@PLGA MS suspension (5&#xa0;mg/kg). Pain thresholds of the left hind paw and cognitive behavioral tests were assessed at specified time points post-administration. After testing, rats were euthanized. Muscle tissue at the injection site was collected for hematoxylin and eosin (H&E) staining to assess inflammatory responses, and hippocampal tissue was harvested for Western blot analysis. The SEM results revealed spherical microspheres with a smooth surface and uniform particle size. FTIR and DSC analyses confirmed stable encapsulation of dezocine (DEZ) in the PLGA matrix, with potential intermolecular interactions and amorphous drug dispersion. In vitro release studies demonstrated a sustained release profile of DEZ from the microspheres. Animal experimental results indicated that the DEZ@PLGA MS group exhibited a significantly prolonged duration of effective analgesia compared to the dezocine injection group. Additionally, DEZ@PLGA MS improved cognitive function, as evidenced by cognitive behavioral tests and Western blot results. H&E staining confirmed no significant inflammatory responses in muscle tissue across all groups, highlighting good biocompatibility of the formulation. Our study demonstrated that DEZ@PLGA MS effectively manages acute postoperative pain over an extended duration and improves postoperative cognitive function.

The frequent occurrences of Babesia microti (B. microti) infection have emerged as a significant global public health safety concern. The diagnosis of patients constitutes a crucial component in the prevention and control of babesiosis, necessitating the urgent establishment of efficient and accurate molecular biology diagnostic methods for B. microti detection. This study aimed to develop a sensitive, specific, and rapid fluorescent recombinase polymerase amplification (RPA) technique for the detection of B. microti (B. microti) and assess its suitability for field use. This study developed and optimized a fluorescent RPA assay targeting the B. microti 18&#xa0;S rRNA gene fragment. The method&#x2019;s sensitivity (detection limit down to 10&#xa0;fg/&#xb5;L genomic DNA), specificity (distinguishing Plasmodium samples), and detection limit for spiked mouse blood samples (across varying parasitemia levels) were evaluated. The detection capability was validated using samples from an infected mouse model (tail-tip blood collected 3&#x2013;21 days post-infection) and two confirmed human cases. Furthermore, 119 blood samples from patients presenting with fever accompanied by thrombocytopenia were tested using both nested PCR and the newly developed fluorescent RPA assay. The agreement between the two methods was analyzed using the Kappa value, and the significance of their difference was assessed using McNemar&#x2019;s test. A fluorescent RPA assay was developed targeting two B. microti 18&#xa0;S rRNA fragments (243&#xa0;bp/191&#xa0;bp). The 191&#xa0;bp fragment demonstrated superior sensitivity (detection limit: 1&#xa0;fg/&#xb5;L genomic DNA) and was selected for assay establishment. Optimized at 39&#xa0;&#xb0;C for 20&#xa0;min, its detection threshold was defined by the lowest detectable concentration (1&#xa0;fg/&#xb5;L). Specificity testing with 8 P. falciparum and 8 P. ovale samples showed fluorescence below this threshold. The assay achieved a detection limit of 0.046 parasites/&#xb5;L blood, demonstrating a 600-fold increase in sensitivity compared to nested PCR, which detects 28.49 parasites/&#xb5;L. In infected mice, both RPA and nested PCR detected parasites by day 9 post-infection (microscopy: day 15), with RPA showing higher positivity. The method confirmed infection in 2 patients. Among 119 febrile patients with thrombocytopenia from Xinyang City, Henan Province, nested PCR detected 20 B. microti-positive cases while fluorescent RPA detected 21. The Kappa value for agreement between the methods was 97.05% (91.31% &#x2212;&#x2009;100%), and McNemar&#x2019;s test indicated no statistically significant difference in their detection rates (S&#x2009;=&#x2009;1, P&#x2009;>&#x2009;0.1). Based on the combined results of both tests, the molecular positivity rate for B. microti in this sample set was 17.65%, with co-infections involving Bunya virus present, accounting for 10.08% of the tested samples. The fluorescence RPA method provides a rapid, sensitive, and specific tool for detecting B. microti and is effective for field screening.

Dentin-cement interface can be considered a critical point in fiber post cementation. Post-space preparation may leave a smear layer and sodium hypochlorite remnants on root dentin, which can compromise bonding between fiber posts and self-adhesive resin cement. This study evaluated the effect of different post-space dentin pretreatment modalities on dentin surface characteristics and push-out bond strength of fiber posts. Forty-five extracted single-rooted mandibular second premolars were endodontically treated and assigned to five groups according to post-space dentin pretreatment (n&#x2009;=&#x2009;9): 2.5% chitosan (CH), 5% apple vinegar (AV), Er,Cr:YSGG laser (ErCr), 970&#xa0;nm diode laser (DL), and 0.9% saline as control group (CG). Fiber posts were cemented using self-adhesive resin cement. Specimens were sectioned into coronal, middle, and apical slices for push-out bond strength testing and failure mode analysis. One additional specimen from each group was examined by scanning electron microscopy (SEM). Push-out bond strength was significantly influenced by surface treatment. The ErCr laser group showed the highest mean bond strength, followed by the chitosan, diode laser, and apple vinegar groups, all of which demonstrated significantly higher values than the control group. Differences between root regions were not statistically significant. Mixed failure was the predominant mode in all experimental groups. SEM revealed superior smear layer removal in the ErCr laser and chitosan groups, whereas the diode laser and apple vinegar groups showed only partial dentinal tubules exposure, and the control group retained a dense smear layer. Post-space dentin pretreatment improved fiber post bond strength and interfacial adaptation compared with saline treatment. ErCr laser and 2.5% chitosan showed the most favorable outcomes and may represent promising conditioning strategies before fiber post cementation.

To evaluate the feasibility and diagnostic performance of ex vivo fluorescence confocal microscopy (FCM) using the Histolog&#xae; Scanner (SamanTree Medical SA, Lausanne, Switzerland) for the assessment of penile cancer (PeCa) specimens, and to compare FCM-based diagnoses with standard formalin-fixed, paraffin-embedded (FFPE) histopathology. We conducted a single-centre study including 12 patients with clinical or radiological suspicion of PeCa who underwent diagnostic or excisional biopsy between June 2022 and November 2023. Fresh biopsy specimens were stained with a nuclear fluorescent dye and scanned ex vivo with the Histolog Scanner. Digital images were retrospectively reviewed by two uropathologists and compared with conventional haematoxylin and eosin-stained FFPE sections. The primary endpoint was diagnostic concordance between FCM and histopathology in detecting invasive squamous cell carcinoma (SCC); secondary endpoints included sensitivity, specificity, and overall feasibility of image acquisition. A total of 29 FCM scans were obtained. Diagnostic image quality was achieved in 28/29 scans (96.6%). FFPE analysis confirmed invasive SCC in eight patients, differentiated penile intraepithelial neoplasia in three, and an atypical squamous proliferative lesion (ASPL) in one. FCM correctly identified tumour category in 11/12 patients, yielding an overall accuracy of 91.7% (95% confidence interval [CI] 61.5-99.8%). Sensitivity for invasive SCC detection was 87.5% (7/8; 95% CI 47.3-99.7%), and specificity was 100% (4/4; 95% CI 39.8-100%). One SCC was misclassified as ASPL on FCM. Median workflow time was ~5&#x2009;min/specimen. Ex vivo FCM is a feasible, rapid imaging technique that enables high concordance with histopathology for the diagnosis of PeCa. This pilot study represents the first assessment of ex vivo FCM in penile malignancies and provides preliminary evidence supporting its potential role in intraoperative margin assessment. Larger prospective studies are required to confirm its diagnostic accuracy and clinical utility.

The olfactory system of parasitoid wasps plays a pivotal role in multiple, essential activities including feeding, mating, oviposition, and host localization. Tetrastichus sp. is a nymphal parasitoid wasp of Agrilus sp. In order to provide morphological basis for further study on behavior of host selection. In the present study, ultrastructure, abundance, distribution and types of the antennal sensilla in both sexes of Tetrastichus sp. were studied using scanning electron microscopy. Antennal sensilla of males and females were compared to those in other hymenopteran parasitoid species and their probable roles were discussed. The antennae of female and male adults of Tetrastichus sp. are similar in shape and belong to geniculate antennae. Antennae consist of radicle, scape, pedicel, anellus, funicle and clava. The female flagellate segment consists of three flagellate subsegments and the male flagellate segment consists of four flagellate subsegments. Six morphologically distinct sensilla types of both sexes of Tetrastichus sp. were observed externally, including B&#xf6;hm sensilla (BBs), sensilla trichodea (St), sensilla chaetica (SCh), sensilla placodea (Sp), Sensilla mammilliformia (SM), and sensilla styloconicum (Ssty). The phenomenon of sexual dimorphism of antennal sensilla is obvious between the male and female, and the morphological structure, types, number and distribution of the sensilla are different, which could be related to the different functions of the antennal sensilla of male and female. This study is of certain significance to provide the morphological and ultrastructural study of antennal sensilla in Tetrastichus sp. and lays a solid foundation for follow-up functional studies.

Curcumin is a polyphenol phytochemical that is extracted from the rhizomes of Curcuma longa Linn. (Zingiberaceae) in the Zingiberaceae family. In recent years, the investigation of curcumin has notably skyrocketed due to its broad pharmacological profile, as evidenced by its antimicrobial, antioxidant, anti-inflammatory, and antitumoral properties. While research and use of curcumin are significantly increasing, its apparent poor water solubility has notably limited its potential as a promising chemical entity. Consequently, efforts to enhance its apparent solubility without altering the chemical properties have been considered. Among the leading techniques is nanocrystasllisation which involves modifying its physicochemical properties, such as particle size, shape and electrical charge. Nanocrystallisation is a nanoscience technique that utilises the drug's crystalline properties to enhance its solubility, bioavailability, and overall pharmacological activity. To date, various nanocrystallisation techniques have been employed, including high-pressure homogenisation, sonoprecipitation, and the solvent-antisolvent precipitation technique. Curcumin nanocrystallisation has often been reported for the improvement of its loading capacity, as it requires fewer excipients and significantly enhances the minimal dose efficiency. Furthermore, the significant role of curcumin in primarily female-associated cancers has been highlighted in some in vitro studies. Therefore, this review will focus on the elaboration of nanocrystals and its promising role in breast and gynaecological cancers. Given the limited studies reported on the anticancer activity of breast and gynaecological cancers curcumin nanocrystals, this review also aims to highlight the research gaps concerning the anticancer activity of curcumin nanocrystals.

The ovarian suspensory ligament (OSL) is routinely manipulated during ovariectomy/ovariohysterectomy in dogs. Despite its surgical relevance, it is traditionally described only macroscopically as a fibrous band, and its microscopic composition has not been characterized. To provide the first detailed histological and ultrastructural characterization of the canine ovarian suspensory ligament. OSL samples were collected from twenty female dogs and processed for histology. Sections were stained with Masson's trichrome, Sirius Red, Weigert's resorcin-fuchsin, and anti-&#x3b1;-smooth muscle actin immunohistochemistry. Tissue components were quantified by surface density using a point-counting method. Scanning electron microscopy was performed to assess ultrastructural organization. Comparisons were made between sides and across age, body weight, and reproductive variables (p&#x202f;<&#x202f;0.05). The OSL did not resemble a dense collagenous ligament. Instead, it showed a heterogeneous architecture dominated by smooth muscle (46.3&#x202f;&#xb1;&#x202f;11.7%), followed by collagenous connective tissue (32.9&#x202f;&#xb1;&#x202f;8.9%), elastic system fibers (5.3&#x202f;&#xb1;&#x202f;2.1%), adipose tissue (2.7&#x202f;&#xb1;&#x202f;2.9%), and blood vessels (2.1&#x202f;&#xb1;&#x202f;2.7%). Immunohistochemistry confirmed the smooth muscle phenotype, and scanning electron microscopy demonstrated a loosely organized collagen network. No significant differences in composition were observed according to side, age, body weight, estrous history, or parity. The canine OSL is not a classical dense fibrous ligament but a muscular ligament. These findings refine its anatomical characterization and have implications for surgical technique and future studies on pharmacologic relaxation.

Microscopic examination (ME) of dermal scrapings is widely used for biological confirmation of cutaneous leishmaniasis (CL), a major health concern in North Africa. This study aims to evaluate this technique and to assess the contribution of stained smears as the DNA source for polymerase chain reaction (PCR). In total, 227 archived Giemsa-stained smears from CL-suspected patients were examined by a standardized graded microscopy followed by scraping, DNA extraction, and kinetoplast DNA real-time PCR amplification. Examination of two 100-mm2 squares for 10 minutes each identified 76 positive slides with varying parasitic loads (1 to >40 amastigotes) and amastigote detection times (range: 1 minute to 18 minutes and 10 seconds). Polymerase chain reaction was more sensitive than ME (99% versus 75.2%), detecting 25 additional positive cases with low estimated parasitic load (median: 2 parasites/slide, interquartile range: 0.5-5.7). This combined approach associates the advantages of microscopy with the increased sensitivity of PCR, being particularly interesting in ME-negative cases with strong clinical suspicion.