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The increased accessibility of soft-tissue data through diffusible iodine-based contrast-enhanced computed tomography (diceCT) enables comparative biologists to increase the taxonomic breadth of their studies with museum specimens. However, it is still unclear how soft-tissue measurements from preserved specimens reflect values from freshly collected specimens and whether diceCT preparation may affect these measurements. Here, we document and evaluate the accuracy of diceCT in museum specimens based on the soft-tissue reconstructions of brains and eyes of five bats. Based on proxies, both brains and eyes were roughly 60% of the estimated original sizes when first imaged. However, these structures did not further shrink significantly over a 4-week staining interval, and 1 week in 2.5% iodine-based solution yielded sufficient contrast for differentiating among soft-tissues. Compared to six “fresh” bat specimens imaged shortly after field collection (not fixed in ethanol), the museum specimens had significantly lower relative volumes of the eyes and brains. Variation in field preparation techniques and conditions, and long-term storage in ethanol may be the primary causes of shrinkage in museum specimens rather than diceCT staining methodology. Identifying reliable tissue-specific correction factors to adjust for the shrinkage now documented in museum specimens requires future work with larger samples.

Forensic examination of the hyoid-larynx complex (HLC) is crucial in diagnosing (inflicted) neck trauma. Clinical Total-Body CT (TBCT) scans of the (non-)calcified HLC lack sensitivity for fracture detection. Micro-computed tomography (micro-CT) is an upcoming imaging modality capable of scanning samples at microscale resolution up to 1 µm. Soft-tissue contrast can be established with iodine staining, known as diffusible iodine-based contrast-enhanced CT (diceCT). We study the additional value of micro- and diceCT in detecting HLC fractures and hemorrhages in forensic cases, compared to standard forensic imaging techniques and findings at autopsy. Fifteen, during forensic autopsy excised, adult HLCs were scanned with micro-CT. Subsequently, the samples were stained with buffered Lugol’s solution (B-Lugol) and scanned with contrast-enhanced micro-CT. The scans were examined on fractures and hemorrhages. Total body CT, optimized clinical CT of the explant and autopsy reports were compared with the micro- and diceCT scans and the results were confirmed using histological examinations. Sixteen potential fractures were identified in the micro-CT scans and with diceCT, nine hyperdensities were detected around potential fractures. Two previously undiscovered vital fractures were found, which were verified by histological examination. This research demonstrated that micro-CT can be a valuable tool to detect fractures of the HLC in forensic neck trauma cases and diceCT enables us to image the injured HLC in detail and indicate possible hemorrhages. With increasingly better stain and scan protocols and more experience gained by forensic radiologists, micro-CT could become invaluable for the examination of HLC trauma. [Display omitted] •Micro- and diceCT provide detailed images of the hyoid larynx complex.•Micro-CT reveals subtle fractures in the hyoid-larynx complex missed by standard CT.•DiceCT imaging can detect hemorrhages, aiding in forensic evaluation of neck trauma.•Two vital fractures, undetected by traditional methods, were confirmed via histology.•Micro-CT and diceCT combined enhance accuracy in forensic neck trauma assessment.

Background To investigate the diagnostic value of conventional white light endoscopy (WLE), narrow band imaging (NBI) endoscopy, and Lugol's iodine staining under WLE (endoscopic iodine staining) in the screening and early diagnosis of nasopharyngeal carcinoma. Methods Patients with nasopharyngeal lesions requiring biopsy attending the Department of Otolaryngology Head and Neck Surgery in our hospital between January 2021 and April 2023 were included in this study. Before biopsy, all subjects underwent conventional WLE, NBI endoscopy, and endoscopic iodine staining. On WLE, according to nasopharyngeal lesion morphology and color, patients were diagnosed with nasopharyngeal carcinoma ( +) or chronic hyperplastic nasopharyngitis (-). On NBI endoscopy, according to nasopharyngeal lesion vascular morphology, patients with type V manifestations (nasopharyngeal carcinoma) were categorized as NBI ( +) and patients with type I-IV manifestations (chronic hyperplastic nasopharyngitis) were categorized as NBI (-). Endoscopic iodine staining (1.6% Lugol's iodine solution) was positive ( +) if the mucosal surface was brown with no white patches, or negative (-) if there was no or light brown staining of the mucosal surface. Patients were divided into 2 groups based on histopathological diagnosis: nasopharyngeal carcinoma or chronic hyperplastic nasopharyngitis. Endoscopic diagnoses were compared with histopathological findings. The diagnostic performance of WLE, NBI endoscopy and endoscopic iodine staining for nasopharyngeal carcinoma were determined. Results This study included 159 patients. On histopathology, 29 patients were diagnosed with nasopharyngeal carcinoma, and 130 patients were diagnosed with chronic hyperplastic nasopharyngitis. There were no significant differences in the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy, and area under the receiver operating characteristic (ROC) curve (AUC) of conventional WLE, NBI endoscopy or endoscopic iodine staining for differentiating nasopharyngeal carcinoma and chronic hyperplastic nasopharyngitis. The diagnostic performance of the combination of conventional WLE, NBI endoscopy and endoscopic iodine staining was significantly improved compared to any procedure alone. Conclusions Conventional WLE, NBI endoscopy or endoscopic iodine staining had good diagnostic performance for differentiating nasopharyngeal carcinoma and chronic hyperplastic nasopharyngitis. In particular, NBI endoscopy and endoscopic iodine staining alone or combined had clinical utility for identifying patients with nasopharyngeal lesions that are eligible for a watch-and-wait strategy.

Purkinje‐related ventricular arrhythmias have been increasingly reported, and with the development of catheter ablation techniques, intervention for Purkinje‐related arrhythmias has been shown to be effective. The characteristics of Purkinje fibres orientation in the 12 canine left ventricles were observed at a gross level by staining the endocardium with Lugol's solution. Purkinje fibres were observed microscopically by HE, Masson's, PAS glycogen, and Cx40 immunohistochemical staining. Staining was successful, and the transverse orientation characteristics of Purkinje fibres were observed by Lugol's staining, and the longitudinal distribution was observed microscopically. The distribution of Purkinje fibres in the canine left ventricle is ‘graded’, ‘layered’, and ‘networked’, which can guide catheter ablation of Purkinje‐related ventricular arrhythmia.

Objective Iodine staining on white light imaging (WLI) is the gold standard for detecting and demarcating esophageal squamous cell carcinoma (ESCC). We examined the effects of texture and color enhancement imaging (TXI) on improving the endoscopic visibility of ESCC under iodine staining. Methods Twenty ESCC lesions that underwent endoscopic submucosal dissection were retrospectively included. The color difference between ESCC and the surrounding mucosa (ΔEe) on WLI, TXI, and narrow‐band imaging was assessed, and ΔEe under 1% iodine staining on WLI and TXI. Furthermore, the visibility grade determined by endoscopists was evaluated on each imaging. Result The median ΔEe was greater on TXI than on WLI (14.53 vs. 10.71, respectively; p < 0.005). Moreover, the median ΔEe on TXI under iodine staining was greater than the median ΔEe on TXI and narrow‐band imaging (39.20 vs. 14.53 vs. 16.42, respectively; p < 0.005 for both). A positive correlation in ΔEe under iodine staining was found between TXI and WLI (correlation coefficient = 0.61, p < 0.01). Moreover, ΔEe under iodine staining on TXI in each lesion was greater than the corresponding ΔEe on WLI. The visibility grade assessed by endoscopists on TXI was also significantly greater than that on WLI under iodine staining (p < 0.01). Conclusions The visibility of ESCC after iodine staining was greater on TXI than on WLI.

Background The practice of endoscopic diagnosis of esophageal squamous cell carcinoma (ESCC) often diverges from evidence-based standards due to various factors, including inadequate dissemination of evidence or a mismatch between evidence and real-world contexts. This survey aimed to identify discrepancies between evidence-based standard practices and community standard practices for ESCC among endoscopists. Methods An online survey targeting endoscopists who perform upper gastrointestinal endoscopy at least weekly was conducted to collect data on clinical practices related to ESCC diagnosis. The survey, comprising 20 questions, was disseminated through multiple professional networks. Descriptive statistical analysis and logistic regression were performed to analyze the data. Results Data from 819 endoscopists were included in the analyses. Notably, a significant proportion employed narrow-band imaging/blue-laser imaging over iodine staining, and preferences varied based on risk assessment for ESCC. In total, 64.0% of endoscopists primarily used iodine solution at a concentration of 1% or less, while 96.5% of endoscopists performed an observation of the oral cavity and the pharynx when conducting upper gastrointestinal endoscopies on individuals at high risk of ESCC. The surveillance interval for metachronous multiple ESCCs was most commonly every 6 months, followed by every 12 months. In addition, most physicians conducted surveillance of metastatic recurrence at 6-month intervals. Conclusions This survey highlights significant gaps between evidence-based and community standard practices in the endoscopic diagnosis of ESCC. These findings underscore the need for enhanced dissemination of evidence-based guidelines and consideration of real-world clinical contexts to bridge these gaps and optimize patient care.

The diagnosis of virus infection can facilitate the effective control of plant viral diseases. To date, serological and molecular methods for the detection of virus infection have been widely used, but these methods have disadvantages if applied for broad-range and large-scale detection. Here, we investigated the effect of infection of several different plant RNA and DNA viruses such as cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), potato virus X (PVX), potato virus Y (PVY) and apple geminivirus on starch content in leaves of Nicotiana benthamiana. Analysis showed that virus infection at an early stage was generally associated with a reduction in starch accumulation. Notably, a reduction in starch accumulation was readily apparent even with a very low virus accumulation detected by RT-PCR. Furthermore, we also observed that the infection of three latent viruses in propagative apple materials was associated with a reduction in starch accumulation levels. Analysis of transcriptional expression showed that some genes encoding enzymes involved in starch biosynthesis were downregulated at the early stage of CMV, TMV, PVX and PVY infections, suggesting that virus infection interferes with starch biosynthesis in plants. Our findings suggest that assessing starch accumulation levels potentially serve as a broad-range indicator for the presence of virus infection.

Reliable, rapid and inexpensive detection of cellulolytic enzymes that can be used for a wide variety of biological and environmental samples are currently in high demand. Here, a new cellulase detection protocol is described that circumvents problems observed with popular agar-based methods by exploiting the ability of carboxymethylcellulose (CMC) to form gel-like surfaces on its own. These pure CMC-layers are sensitive to cellulolytic degradation and stainable by Gram’s iodine without showing unwelcome reactions with other enzymes. The staining intensity negatively correlates with the enzyme activity and can be used for quantification. Cellulase activities are not obstructed by high sugar contents (e.g., in plant material) which limit the applicability of other quantification methods, making our new method particularly attractive for screening of plant extracts. A useful variant of this new method is its applicability to plant tissue prints for spatial mapping of the cellulolytic activity in a zymogram-like fashion.

Background In cases of Hirschsprung disease, complete and reproducible resection of the aganglionic bowel is ideal to achieve good postoperative bowel function. Reliable identification of the upper margin of the surgical anal canal, which is the squamous-columnar junction, is necessary during transanal pull-through. Here, we describe a novel staining technique using Lugol’s iodine stain to visualize the upper margin of the surgical anal canal. Methods Lugol’s iodine staining was performed in five patients with Hirschsprung disease treated using a single-stage laparoscopic transanal pull-through modified Swenson procedure. In two of these patients, endocytoscopic observation with ultra-high magnification was performed using methylene blue and crystal violet to mark the border of the squamous epithelium at 1 week before surgery. The alignment between the incisional line, which was revealed using Lugol’s iodine staining and endocytoscopic marking, was evaluated. Complications, including postoperative bowel dysfunction, were evaluated. Results In all cases, Lugol’s iodine staining produced a well-demarcated line. The endocytoscopic marking of the upper margin of the surgical anal canal was aligned with the line revealed by Lugol’s iodine staining. There were no complications associated with the transanal pull-through procedure, including postoperative bowel dysfunction. Conclusions Lugol’s iodine staining could be a safe and practical method to visualize the upper margin of the surgical anal canal intraoperatively. This finding may be useful for surgeons to make a consistent removal of the aganglionic bowel during surgery for Hirschsprung disease.

PurposeThis study aimed to introduce a novel approach to study the facial mimetic muscles (FMMs) in relation to the nasolabial fold (NLF) and realize the visualization of complex three-dimensional (3D) structures and spatial relationships of the FMMs.Materials and MethodsNano-computed tomography (nano-CT) and iodine staining techniques were used to obtain the two-dimensional (2D) radiographs of the FMMs. Materialise Mimics software was then used to reconstruct the 3D model of the FMMs.ResultsThe zygomaticus major muscle (ZMM) was divided into trunk fibers and branch fibers. The trunk fibers of the ZMM were subdivided into branch fibers layer-by-layer. Adipose tissue in the cheek was not a mass of unorganized fat. It was separated and fixed by branch fibers. Moreover, the trunk fibers of the ZMM were directly connected to the levator anguli oris (LAO), not the skin. On the contrary, the ZMM was connected to the skin by its subdivided branch fibers indirectly. The muscle fibers in the modiolus were organized, rather than disorganized. In other words, the terminal of the trunk fibers of the ZMM was located in the LAO. Moreover, the terminal of the trunk fibers of the LAO was located at the terminal of the trunk fibers of the musculus depressor anguli oris at the corner of the mouth. Furthermore, the levator labii superioris alaeque nasi was not directly connected to the orbicularis oris muscle. It was connected to the combination of the LLS and the rhinaeus.ConclusionsAlthough nano-CT has many disadvantages, it enabled the 3D anatomical study of the FMMs in relation to the NLF when combined with iodine staining. We imported the 2D images obtained by nano-CT scanning into the Mimics software, successfully reconstructed the FMMs, and finally obtained images of complex 3D structures of the FMMs. The shapes, positions, and 3D spatial relationships of the FMMs were clearly visualized. The novel insights into the 3D anatomy of the FMMs may help understand the formation of the NLF. Finally, the results of this study may help improve the rejuvenation surgery of the NLF soon.Level of Evidence IVThis journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.