Histological Sections Research Articles

7817 A Rare Case of Intrathyroidal Thyroglossal Duct Cyst Presenting as a Rapidly Expanding Left Thyroid Goiter

Abstract Disclosure: C.M. Godar: None. A.J. Spiro: None. K.F. Brown: None. M.K. Shakir: None. T.D. Hoang: None. Introduction: Thyroglossal duct cysts are the most common congenital cyst in the head and neck, found during childhood as a midline neck mass. We present a rare case of a large intrathyroidal thyroglossal duct cyst with left lateral extension in an adult patient. Case Report: A 50-year-old man with family history of familial adenomatous polyposis presented for thyroid nodule evaluation. Thyroid ultrasound for screening of thyroid cancer showed a 4.8 x 4.5 x 2.5cm mixed cystic/solid hyperechoic thyroid nodule with smooth margins in the left lower thyroid lobe. He reported no compressive symptoms. FNA with standard 27G needle was insufficient for evaluation with Bethesda I result. Attempted syringe aspiration did not produce cystic fluid for analysis. The patient returned to clinic 8 weeks later due to visible enlargement of his anterior neck and new neck pressure with swallowing. Physical exam was significant for a new, firm palpable mass over the left thyroid isthmus. Bedside ultrasound noted expansion of the previously visualized mass now extending into the thyroid isthmus. FNA was reattempted with a 25G needle, and cytologic material was again inadequate. He elected to undergo left hemithyroidectomy. Histologic sections demonstrated a 3.4cm simple squamous, non-keratinizing cyst lining overlying a thickened fibrotic wall with mixed acute and chronic inflammation surrounded by benign thyroid tissue, consistent with an intrathyroidal thyroglossal duct cyst. Discussion: The thyroglossal duct derives from epithelial tissue during embryogenesis and atrophies during weeks 8-10 of gestation. A persistent duct may lead to cyst formation in the presence of remnant epithelial tissue secretions and local neck inflammation. A thyroglossal duct cyst is usually located midline in the anterior neck. This stands in comparison to a branchial cleft cyst, which arises due to failure of atrophy of the second branchial groove during embryogenesis and forms a cyst that is usually located in the upper lateral neck compartment. Other common neck masses include lipomas, dermoid cysts, or ectopic thyroid tissue. This case highlights the rarity of intrathyroidal thyroglossal duct cyst in an adult patient and the importance of definitive diagnosis and appropriate management. Presentation: 6/2/2024

Reconstructing infant mortality in Iberian Iron Age populations from tooth histology

The Neonatal Line (NNL) of the tooth serves as a unique identifier, allowing us to distinguish whether a child survived birth. This line is essential for assessing the age at death of infants from skeletal remains found in archaeological contexts. Our primary objective is to accurately determine the age of infant intramural inhumations from the Iberian Iron Age (8th-1st centuries BC) by analyzing histological sections of tooth germs. Due to their fragility and high susceptibility to taphonomic factors, these samples are challenging to handle. By accurately assessing their age, we aim to classify individuals into various stages of infant mortality, which will help reconstruct infant mortality patterns in these populations.We analyze unerupted and still-forming crown deciduous teeth from 45 infant burials. By calculating Crown Formation Time (CFT) and identifying the NNL, we determine both gestational and chronological ages. We further validate the reliability of NNL identification through Synchrotron X-ray Fluorescence (SXRF) elemental analysis (Ca, Zn, Cu) on two contemporary and two archaeological samples.Our histological study reveals the chronological age of 38 infants from Iberian settlements, ranging from the 30th week of gestation to the 2nd postnatal month. The age distribution shows an attritional mortality pattern, with nearly half experiencing perinatal mortality, including preterm births. These findings support the hypothesis that mortality was primarily attributed to natural causes. Our research enhances the understanding of infant life history events in prehistory by combining histological analysis of tooth NNL and CFT, highlighting the technique's potential and limitations.

Radiographic, computed tomographic, and histologic characteristics of bone for clinically normal laying hens in a free-range housing system.

Laying hens are increasingly being kept in backyard flocks and considered family pets; however, diagnostic imaging characteristics of bone for clinically normal backyard hens are currently limited. This prospective, descriptive study was to describe radiographic, computed tomographic, and histologic characteristics of bone for a group of clinically normal laying hens housed in conditions comparable to those of backyard flocks. Sixteen 60-week-old Lohmann Brown laying hens were included. Hens were housed in a free-range unit with outdoor access at a university research and teaching farm. Hens were defined as clinically normal by the farm manager and a veterinary researcher in laying hen behavior and welfare. Findings from the horizontal beam, left lateral, sternal radiographs (n=16), postmortem, and whole-body CT scans (n=4) were recorded by a veterinary radiologist and a research technician. Histologic findings for sternal, femoral, and tibiotarsal bone samples (n=5) were recorded by a veterinary pathologist. The most frequent radiographic findings for the sternal carina (keel bone) were smoothly marginated concave deviations of the ventral margin and caudal section fractures. Multiple punctate mineral opacities (PMOs) were present in radiographs and CT images for all hens and were involved in the sternal carina and multiple other bones in the axial and appendicular skeleton. No bone abnormalities were identified in any histologic sections where PMOs were radiographically detected. Authors propose that PMOs are normal radiographic and CT findings in the bones of mature, laying hens and may represent temporary calcium reservoirs formed during osteoclastic activities.

Tumor microenvironment characteristics association with clinical outcome in patients with resected intestinal-type gastric cancer.

Tumor microenvironment (TME) characteristics including tumor stroma ratio (TSR), tumor budding (TB), and tumor-infiltrating lymphocytes (TILs) were examined in resected gastric cancer. These TME features have been shown to indicate metastatic potential in colon cancer, and intestinal-type gastric cancer (IGC) has pathological similarities with that malignancy. TSR, TB, and TILs were quantified in routine histological sections from 493 patients with IGC who underwent radical resection at 2 university hospitals in China from 2010 to 2016. TME variables were dichotomized as follows: TSR (50%), TILs (median), TB per international guidelines (4 buds/0.785mm2), and platelet-lymphocyte ratio (PLR) per survival ROC. Association of TME features with patient clinicopathological characteristics, time-to-recurrence (TTR), and cancer-specific-survival (CSS) were examined using univariate and multivariate analysis, including a relative contribution analysis by Cox regression. Patients whose tumors showed high TSR or high TB or low TILs were each significantly associated with increased T and N stage, higher histological grade, and poorer TTR and CSS at 5 years. Only TSR and N stage were independently associated with TTR and CSS after adjustment for covariates. PLR was only independently associated with TTR after adjustment for covariates. Among the variables examined, only TSR was significantly associated with both TTR (HR 1.72, 95% CI, 1.14-2.60, P = .01) and CSS (HR 1.62, 95% CI, 1.05-2.51, P = .03) multivariately. Relative contribution to TTR revealed that the top 3 contributors were N stage (45.1%), TSR (22.5%), and PLR (12.9%), while the top 3 contributors to CSS were N stage (59.9%), TSR (14.7%), and PLR (10.9%). Among the examined TME features, TSR was the most robust for prognostication and was significantly associated with both TTR and CSS. Furthermore, the relative contribution of TSR to patient TTR and CSS was second only to nodal status.

A-289 Multiple Gastric Polyps as an Unusual Presentation of Syphilitic Gastritis

Abstract Background Syphilis is a sexually transmitted infection caused by Treponema pallidum and can clinically progress through four different stages. Syphilitic gastritis is one of the rare manifestations of the disease, harboring the potential to mimic several types of unrelated disorders such as malignant lymphomas, Langerhans cell histiocytosis, auto-immune gastritis and refractory Helicobacter pylori infection. The classics symptoms are weight loss, abdominal pain, and gastroparesis, nausea, vomiting and hematemesis. The most common endoscopic finding is gastric ulcers. The rising trends of syphilis in the last decade, detected in both developing and developed countries, calls for continued efforts for case detection on rare syphilis manifestations. Methods A 35-year-old male patient was submitted to gastric biopsy due to endoscopic findings of multiple polyps in the corpus and antrum of stomach. The material was processed and analyzed in a pathology laboratory in São Paulo, Brazil. Histological sections were stained with hematoxylin and eosin and immunohistochemical staining. Results The specimens exhibited intense inflammatory lympho-histiocytic infiltrate, rich in plasma cells, neutrophils and eosinophils affecting the gastric mucosa. Immunohistochemical staining revealed spiral-shaped bacteria consistent with T. pallidum affecting the gastric mucosa surface, adjacent to vessels, and permeating glands. Conclusions This case report has the goal of showing the occurrence of syphilitic gastritis, an uncommon extracutaneous manifestation of T. pallidum infection with the potential to mimic other etiologies. Due to the rarity of syphilitic gastritis and its non-specific symptoms, a combination of clinical, endoscopic, and histopathological findings is necessary to secure diagnosis of syphilitic gastritis. The advent of immunohistochemistry for T. pallidum has facilitated microscopic diagnosis, as special stains such as Warthin-Starry can still pose interpretation challenges.

Histological Validation of 3D Variable Flip Angle TSE Multi-Contrast Magnetic Resonance Vessel Wall Imaging in Characterizing Carotid Vulnerable Atherosclerotic Plaques

BackgroundAccurate assessment of the vulnerability of carotid atherosclerotic plaques is crucial for stroke prevention. The three-dimensional (3D) magnetic resonance (MR) vessel wall imaging (VWI) has been increasingly employed to evaluate carotid plaques due to its extensive coverage and isotropic high spatial resolution. However, the accuracy of such technique lacks validation by histology. ObjectiveThis study aims to validate the accuracy of 3D multi-contrast MR VWI used variable-flip-angle (VFA) and turbo spin echo (TSE) readout in identifying vulnerable carotid plaques, using histological analysis as a reference. MethodsTwenty-one male patients (mean age: 64.4 ± 7.2 years) scheduled for carotid endarterectomy (CEA) were recruited for this study. All patients underwent carotid multi-contrast MR VWI, including 3D T1- and T2-weighted variable flip angle-based turbo spin echo (VFA-TSE) sequences, as well as 3D time of flight (TOF) MR angiography (MRA), using a 3.0T MR system. Histological processing was performed for carotid plaque specimens. The presence or absence, along with the area measurements, of lipid-rich necrotic core (LRNC), intraplaque hemorrhage (IPH), and calcifications (CA) were independently evaluated on both MR images and histological sections. Cohen’s kappa (κ) analysis was utilized to determine the agreement between 3D multi-contrast MR VWI and histology in identifying carotid plaque compositions before and after excluding compositions bellow certain size threshold. Spearman’s correlation analysis was also conducted to assess the agreement in quantifying plaque compositions. ResultsA total of 81 slices of MR images were successfully matched with histological sections. Moderate to almost perfect agreements were observed between 3D MR VWI and histology in the identification of LRNC (κ: 0.85 and 0.89), IPH (κ: 0.65 and 0.69), and CA (κ: 0.46 and 0.62) before and after excluding compositions smaller than 0.79 mm2. Strong to very strong correlations were found in the quantification of plaque compositions including LRNC (r=0.88), IPH (r=0.80), and CA (r=0.74) between MR imaging and histology. ConclusionThe 3D VFA-TSE multi-contrast MR VWI is capable of accurately characterizing vulnerable carotid atherosclerotic plaques.

Telomere Length Measurement in Human Tissue Sections by Quantitative Fluorescence In Situ Hybridization (Q-FISH).

Telomeres in most somatic cells shorten with each cell division, and critically short telomeres lead to cellular dysfunction, cell cycle arrest, and senescence. Thus, telomere shortening is an important hallmark of human cellular senescence. Quantitative fluorescence in situ hybridization (Q-FISH) using formalin-fixed paraffin-embedded (FFPE) tissue sections allows the estimation of telomere lengths in individual cells in histological sections. In our Q-FISH method, fluorescently labelled peptide nucleic acid (PNA) probes are hybridized to telomeric and centromeric sequences in FFPE human tissue sections, and relative telomere lengths (telomere signal intensities relative to centromere signal intensities) are measured. This chapter describes our Q-FISH protocols for assessing relative telomere lengths in FFPE human tissue sections.

Effect of Justicia insularis Leaf Extract and Fractions on Oxidative Stress Markers, Liver Function Parameters and Liver Histology of Plasmodium berghei -Infected Mice

Justicia insularis (Family-Acanthaceae) is used in Ibibio ethnomedicine to treat malaria. The leaf extract and fractions of J. insularis were investigated for antioxidative stress and hepatoprotective activities in Plasmodium berghei-infected mice. The leaf extract (100-300 mg/kg, p.o.) exerted significant (p<0.05) antimalarial activity against P. berghei infection in curative test with ethyl acetate fraction demonstrating the highest activity. The extract/fractions treatment caused significant (p<0.05) reductions in liver enzymes (ALT, AST and ALP), total and conjugated bilirubin of the treated infected mice and also decreased significantly (p<0.05) total protein and albumin levels of the treated mice relative to control. The leaf extract and fractions further improved significantly (p<0.05) the levels of oxidative stress markers enzymes and molecules (CAT, GPx, GST, SOD) of the treated infected mice with no significant (p>0.05) effect on GSH. The MDA levels in the livers of the treated infected mice were significantly (p<0.05) reduced relative to control. Histology of liver sections revealed absence or significant reductions in pathological features in infected mice treated with leaf extract (100 mg/kg), DCM and ethyl acetate fractions compared to untreated infected mice. These results suggest that the leaf extract/fractions of Justicia insularis possess antioxidative stress and hepatoprotective potentials, which is an added advantage to its antimalarial property.

Fat Embolism Does Not Alter Cardiac Structure or Induce Pathological Changes in a Rat Model

IntroductionFat embolism (FE) encompasses conditions in which fatty substance becomes embedded in a tissue/organ. Fat emboli most commonly affect the lungs in a trauma setting. This can lead to both significant pathology locally and systemically including changes in structure, inflammatory response, activation of the renin-angiotensin system, and subsequent hypoxia. In fact, changes in skin, brain, lungs, and kidneys have been noted in FE syndrome. Because there is an extensive record of pathology reports on this condition without evidence of direct cardiac involvement, as well as our studies showing apparent complete recovery after the acute embolism, we hypothesized that structural changes similar to the lung and at the same time course would not be observed in the heart. MethodsWe used a rat model of FE previously described by our group where we have documented significant lung pathology. In this study, we analyzed both pulmonary and cardiac structure, histology, and gene expression at 48 h and 10 wks post fat injection to mimic FE. ResultsDespite severe inflammatory evidence and structural changes to the lung and vasculature up to 10 wks after FE, we found no significant alterations to cardiovascular morphometry including lumen patency ratio, adventitia/media ratio, fibrosis content, and heart chamber/wall dimensions in stained histological sections. Additionally, genetic markers of cardiac pathological hypertrophy were not significantly elevated 48 h or 10 wks after fat treatment. Oil Red O staining showed increased fat droplet content within lung and aorta tissue, but not in the myocardium. ConclusionsOur study suggests that, in contrast to the lungs, the heart is more resistant to the inflammatory and remodeling responses that result from FE, possibly due to the organ-specific differences in fat retention.

The potential of x-ray virtual histology in the diagnosis of skin tumors.

Histopathological analysis represents the gold standard in clinical practice for diagnosing skin neoplasms. While the current diagnostic workflow has specialized in producing robust and accurate results, interpreting tissue architecture and malignant cellular morphology correctly remains one of the greatest challenges for pathologists. This paper aims to explore the prospect of applying x-ray virtual histology to human skin tumor excisions and correlating it with the histological validation. Seven skin biopsies containing intriguing melanoma types and pigmented skin lesions were scanned using x-ray Computed micro-Tomography (μCT) and then sectioned for conventional histology assessment. The tissue microarchitecture reconstructed by μCT offers detailed insights into diagnosing the malignancy or benignity of the skin lesions. Three-dimensional reconstruction via x-ray virtual histology reveals infiltrative patterns in basal cell carcinoma and evaluated invasiveness in melanoma. The technology enables the identification of pagetoid distributions of neoplastic cells and the assessment of melanoma depth in three dimensions. Although the proposed approach is not intended to replace conventional histology, the non-destructive nature of the sample and the clarity provided by virtual inspection demonstrate the promising impact of μCT as a valid support method prior to conventional histological sectioning. Indeed, μCT images can suggest the optimal sectioning position before using a microtome, as is commonly performed in histological practice. Moreover, the three-dimensional nature of the proposed approach paves the way for a more accurate assessment of significant prognostic factors in melanoma, such as Breslow thickness, by considering the whole micro-volume rather than a two-dimensional observation.

Colonic targeting insulin-loaded trimethyl chitosan nanoparticles coated pectin for oral delivery: In vitro and In vivo studies

Colon-targeted delivery offers several benefits for oral protein delivery, such as low proteolytic enzyme activity, natural pH, and extended residence time, which improve the bioavailability of the encapsulated protein. Therefore, we hypothesize that developing a novel colonic nanocarrier system based on modified chitosan, which enhances solubility at natural pH and is coated with a colon-degradable polymer, will provide an effective delivery system for oral insulin. This study aims to synthesize insulin-loaded pectin-trimethyl chitosan nanoparticles (Ins-P-TMC-NPs) as an oral insulin delivery system and evaluate their efficacy both in vitro and in streptozotocin-induced diabetic male rats. N-trimethyl chitosan (TMC), synthesized via a methylation method, was used to prepare insulin-TMC nanoparticles coated with pectin via the ionic gelation method. The nanoparticles were characterized for their physicochemical properties, cumulative release profile, and surface morphology. In vitro biological cytotoxicity and cellular uptake of the nanoparticles were evaluated against HT-29 cells using an MTT assay and fluorescent microscopy, respectively. The in vivo blood glucose-lowering effect was assessed in diabetic male Sprague–Dawley rats, with in vivo toxicity evaluated in the liver, spleen, duodenum, pancreas, and kidney through histological studies (H&E staining). The results showed that Ins-P-TMC-NPs were spherical, with an average size of 379.40 ± 40.26 nm, a polydispersity index of 24.10 ± 1.03 %, a zeta potential of +17.20 ± 0.52 mV, and a loading efficiency of 83.21 ± 1.23 %. Compared to uncoated TMC nanoparticles, Ins-P-TMC-NPs reduced insulin loss in simulated gastrointestinal fluid by approximately 67.23 ± 0.97 % and provided controlled insulin release in simulated colonic fluid. In vitro bioactivity studies revealed that Ins-P-TMC-NPs were non-toxic, with cell viability of 91.12 ± 0.91 %, and exhibited high cellular uptake in the HT-29 cell line with a fluorescence intensity of 37.80 ± 2.40. Furthermore, in vivo studies demonstrated a sustained reduction in blood glucose levels after oral administration, peaking after 8 h with a glucose reduction of 87 ± 1.03 %. Histological sections showed no signs of toxicity. Overall, the developed colon-targeted oral insulin delivery system shows great potential as a candidate for oral insulin administration.

Validation of Metallothionein Immunohistochemistry as a Highly Sensitive Screening Test for Wilson Disease

Wilson’s disease (WD) is a rare autosomal recessive condition with protean clinical manifestations that results from biallelic ATP7B mutations. However, non-destructive tissue tests to be applied clinically to tissue specimens are not widely available to effectively assess patients for possible WD. Previously, we showed that metallothionein (MTH) immunohistochemistry has a high sensitivity and specificity for WD diagnosis and, thus, represents a potentially powerful diagnostic tool that can be used in routine histologic sections. We sought to validate this finding in a large cohort of bona fide WD patients and to correlate metallothionein expression with other histologic features.We identified 91 cases of WD, which included 28 needle biopsies and 64 explants from 14 centers worldwide. Histologic features were evaluated, and a histopathological pattern was assigned to each case. All cases were evaluated with Masson trichrome and MTH immunohistochemistry (clone UC1MT, Abcam) using previously published technique. Liver tissues from chronic cholestatic diseases (n= 42) were used as controls.The median age of the cohort was 28.5 years. Out of 91 total cases, 83 were positive for MTH immunostain. In the controls, all 42 cases were negative for MTH immunostain. The sensitivity and specificity of MTH immunostain for WD were 91.20% and 100%, respectively.MTH immunohistochemistry is a highly sensitive and specific cost-effective screening tool for WD. It can be used for patients across age groups, varied histologic patterns, and fibrosis stages. This marker could prove to be a valuable tool in the evaluation of patients with possible WD.

Mineralization of equine proximal sesamoid bones precedes articular cartilage and fibrocartilaginous enthesis maturation in early postgestational development.

To describe the development and maturation of equine proximal sesamoid bones (PSBs) in fetuses and young horses using radiography, microcomputed (micro)-CT, and histology. A descriptive study. Forelimb PSBs from 12 equids ranging in age from 105 days of gestation to 540 days postgestation were evaluated. Radiography was used for preliminary assessment of metacarpophalangeal joint and PSB mineralization, and micro-CT imaging was performed to assess mineralized PSBs. Tissue volume, bone volume fraction, height, width, depth, trabecular thickness, and anisotropy were quantified from midplanar micro-CT sections in 3 dimensions. Midsagittal PSB histologic sections stained with H&E and Safranin O/Fast Green were used to determine the ratio of ossification center to cartilage template size and to describe the formation and development of the cartilage template, ossification center, spherical growth plate, articular cartilage, and entheses. Mineralization of equine PSBs is associated with cartilage canals and a spherical growth plate that undergoes endochondral ossification during the late gestation to early postgestational period. The apical, flexor, basilar, and articular ossification fronts demonstrate morphologic variability. Structural organization of the articular cartilage and entheses occurs concurrently with the development of an underlying plate of compact bone. At 540 days postgestation, the fibrocartilaginous entheses of the flexor cortex of the PSB had yet to mature. Equine PSBs mineralize predominantly by endochondral ossification during the late gestation to early postgestational period. Mineralization precedes maturation of the articular cartilage and fibrocartilaginous entheses. The postgestational maturation of the PSB and its surrounding tissues may predispose young horses to developing lesions at these sites, such as apical avulsion fractures, warranting further investigation.

Coated and Uncoated Magnetic Iron Oxide Nanoparticles: Toxicity Assessment and Properties of Non Accumulation in Biological Tissues

Background: There are an increasing number of studies on magnetic nanofeeders with multiple applications in agriculture and food processing. Among the magnetic separation oxides, iron oxide has emerged as an indispensable tool in nanoscale differentiation, especially bio-nanoscale. This is attributed to the characteristics of such as size, shape, magnetism and coexistence. Aim of the study: The aim of this study feeding experiment is to prove the non-toxicity of magnetic nano-iron oxides, whether coated when dosed with distilled water or uncoated in milk or distilled water. Subject, materials and methods: Materials: magnetic iron oxide nanoparticles was obtained from SkySpring Nanomaterials (U.S.A.) at a size of 30 nm. Acacia gum (Gum Arabic) is prepared from Thomas Baker (Indian), Sodium hydroxide from Riedel-de Haën (Germany), Potassium phosphate dibasic from CDH (Indian), Potassium phosphate mono basic from HIMEDIA (Indian),Potassium bromide from AVONCHEM(UK), MNPs are characterized by various tools to examine their physicochemical properties. The size of NPs plays a major role in exhibiting different physicochemical properties, and even a small difference in their nanoscale dimensions can alter their properties. Some of the instruments used for their characterization are Atomic Force Microscopy (AFM), Energy Dispersive X-ray Diffraction (EDXD), Field-Emission Scanning Electron Microscopy (FE-SEM), Fourier Transform Infrared Spectrometer. Results: After the completion of the experimental period for this study (28 days), the laboratory animals were sacrificed using ether anesthesia. The animals were then dissected using dissection tools to remove these organs. The organs were weighed using a sensitive balance after being washed with 0.9% normal saline solution and dried with filter paper [1][2]. The liver, kidneysamples were collected in pre-labeled plastic containers and preserved in 10% formalin until histological sectioning could be performed. Tissue sections were performed for all organs - liver, kidney and they were placed under a microscope. The results showed no damage to these organs. Conclusion: The findings suggest that both arabic gum-coated and milk-coated magnetic iron oxide nanoparticles, at the tested doses and treatment durations, did not induce observable tissue damage or pathological alterations in the kidneys or liver of the treated rats.

Investigation of Sonication Parameters for Large-Volume Focused Ultrasound-Mediated Blood-Brain Barrier Permeability Enhancement Using a Clinical-Prototype Hemispherical Phased Array.

Background/Objectives: Focused ultrasound (FUS) and microbubble (MB) exposure is a promising technique for targeted drug delivery to the brain; however, refinement of protocols suitable for large-volume treatments in a clinical setting remains underexplored. Methods: Here, the impacts of various sonication parameters on blood-brain barrier (BBB) permeability enhancement and tissue damage were explored in rabbits using a clinical-prototype hemispherical phased array developed in-house, with real-time 3D MB cavitation imaging for exposure calibration. Initial experiments revealed that continuous manual agitation of MBs during infusion resulted in greater gadolinium (Gd) extravasation compared to gravity drip infusion. Subsequent experiments used low-dose MB infusion with continuous agitation and a low burst repetition frequency (0.2 Hz) to mimic conditions amenable to long-duration clinical treatments. Results: Key sonication parameters-target level (proportional to peak negative pressure), number of bursts, and burst length-significantly affected BBB permeability enhancement, with all parameters displaying a positive relationship with relative Gd contrast enhancement (p < 0.01). Even at high levels of BBB permeability enhancement, tissue damage was minimal, with low occurrences of hypointensities on T2*-weighted MRI. When accounting for relative Gd contrast enhancement, burst length had a significant impact on red blood cell extravasation detected in histological sections, with 1 ms bursts producing significantly greater levels compared to 10 ms bursts (p = 0.03), potentially due to the higher pressure levels required to generate equal levels of BBB permeability enhancement. Additionally, albumin and IgG extravasation correlated strongly with relative Gd contrast enhancement across sonication parameters, suggesting that protein extravasation can be predicted from non-invasive imaging. Conclusions: These findings contribute to the development of safer and more effective clinical protocols for FUS + MB exposure, potentially improving the efficacy of the approach.

POLARIZATION-CORRELATION MAPPING OF MICROSCOPIC IMAGES OF BIOLOGICAL TISSUES OF DIFFERENT MORPHOLOGICAL STRUCTURE

The results of polarization-correlation mapping of the fourth parameter of the two-point Stokes vector of microscopic images of histological sections of biological tissues with different architectonics of the optically anisotropic polycrystalline component are presented. The coordinate distributions of randomly generated values representing the modulus of the fourth parameter of the polarization-correlation vector from microscopic images of histological sections of fibrillar tissues (such as skin dermis) and parenchymal tissues (like spleen) have been replicated. The statistical analysis results of algorithmically generated coordinate distributions of random values representing the modulus of the fourth parameter of the polarization-correlation vector from microscopic images of histological sections of biological tissues with varying morphological structures are provided.

Changes in masseter muscle structure, membrane lipid peroxidation and Ca-ATPase activity as effects of different local anesthetics.

Local anesthetics (LA) can cause undesired effects such as sustained contraction of skeletal muscles as a result of structural and functional changes. Proper skeletal muscle function is controlled by intracellular Ca2+ concentration and efficient energy (ATP) production, which is closely related to cell ultrastructure. Aim: The aim of this study was to identify the structural and functional changes caused by LAs. Materials and Method: Male Wistar rats weighing 200 to 250g were used (n:49). They were divided into seven groups. One group was not anesthetized or treated (Control). The other six groups underwent intramuscular (IM) anesthesia with xylazine 2% (0.05 ml) and ketamine 50 mg/ml (0.1 ml/100g rat weight), and one of the following was applied to the masseter muscle (MM): no further treatment (Anesthetic Control group, CA); 0.1ml physiological saline solution (group SF); Carrageenin (group Carr) 1% as positive control group; prilocaine (group Pri), mepivacaine (group Mepi); or articaine (group Arti) 0.3M, IM. The animals were euthanized by cervical dislocation one hour after treatment. The effects of the different anesthetics on the MM were evaluated histologically and by electronic microscopy (EM). Ca-ATPase and membrane lipid peroxidation (LPX) were evaluated in muscle homogenates under the same conditions as those used to prepare the histological sections. Results: In general, structural damage and increased muscle contraction were observed in tissues treated with anesthetics. The most extreme values of Ca-ATPase activity and LPX were observed in the positive control group (carrageenin). Results were analyzed by one-way ANOVA for multiple comparisons and Tukey's test (p < 0.05). Conclusions: The results suggest that in the short term, local anesthetics affect the muscle function and are associated to structural changes.

Use of Carrot Oil, Olive Oil, Pine Oil, Rose Oil in Comparison to Xylene as Clearing Agent During Tissue Processing: an in Vitro Study

ABSTRACT: Background: Clearing plays a vital role in histological section preparation. Xylene has long been valued for its ability to efficiently deparaffinize and clear tissue samples. However, its hazardous properties have raised health and environmental concerns, leading researchers to seek alternative solvents for tissue processing. This study utilized safer substitutes to address these concerns. Aim: To assess the clearing ability of four different oils i.e., Carrot oil, Olive oil, Pine oil, and Rose oil in comparison with that of xylene. Materials and Methods: Thirty different formalin-fixed tissue samples were cut into five parts and cleared using Carrot oil, Olive oil, Pine oil, Rose oil, and xylene followed by routine processing, sectioning, and staining steps. Presence or absence of shrinkage, ease of cutting, nucleus, cytoplasm details and staining quality were assesed and comparison was done between each of them cleared with the 4 different oils with that cleared using xylene. Results: The results showed that all four oils cleared tissues as effectively as xylene, with Pine oil superior in nucleus and cytoplasm details, as well as staining quality insignificant number of samples. Conclusion: These oils not only more affordable and eco-friendly than xylene but can also serve as effective clearing agents.

Clinical and histological study of cold physical plasma jet in treatment of full thickness skin wounds of normal and diabetic dogs.

To study the effect of physical plasma jet in treating open wounds in diabetic and non-diabetic dogs. The experimental study was conducted at the College of Veterinary Medicine, University of Baghdad, Iraq, from 20 January, 2020 to 1st May 2020, according to (no. 1364/ P.G), and comprised adult male diabetic and nondiabetic dogs. They were divided into non-diabetic group N and diabetic group D. Each group was further divided into treatment subgroup T and control subgroup C. Each dog was subjected to 4 wounds 3×3cm in size. Homemade helium non-equilibrium atmospheric pressure plasma jet therapy was used for healing purposes. Clinical parameters were evaluated by observation, while histological images were scored based on the semi-quantitative evaluation of histological sections on days 3-, 7- and 21-days post-wound. Data was analysed using SPSS Version 26 with One-way ANOVA for statistical analysis between groups and sub-groups. Of the 24 dogs, 12(50%) were in each of the two groups, which were further divided into subgroups having 6(50%) dogs each. The therapy accelerated the process of wound healing in the NT and DT subgroups compared to NC and DC. Clinical observations revealed early and complete closure of plasma-treated wounds in NT and DT subgroups started at day 30 post-wounding, while in the NC subgroup it started at day 35, and in the DC subgroup, wounds failed to close even after 35 days post-wounding. Histological analysis suggested that a plasma jet supported epithelisation, angiogenesis, formation of new hair follicles and collagen fibres, while it also controlled inflammation. In addition, in NT and DT subgroups, it increased the proliferation of fibroblasts and deposition of collagen, and there is a very, highly significant difference between groups (P<0.001), and a significant difference between days (P<0.05). Home-made helium non-equilibrium atmospheric pressure plasma jet therapy improved the quality and pace of wound healing.

Experimental and Histological Validation of Nanostructured Products Use in Maxillofacial Surgery

The aim of the study is using experimental histological techniques to validate the beneficial potential of specific medical devices made of nanostructured titanium, and compare their effect with factorymade products currently applied in maxillofacial surgery. Material and methods. The study involved 50 sexually mature male Chinchilla rabbits. An open fracture of the lower jaw was simulated in the animals. In the 1st series of the experiment, the fragments were fixed using nanostructured miniplates and miniscrews made of Grade 4 titanium. In the 2nd series, they were fixed with standard miniplates and miniscrews from Konmet LLC. The material for the study was taken on the 7th, 14th, 21st, 28th and 40th days after the operation. In all series the object of the study was the extirpated lower jaw in the fracture area. The prepared histological sections were stained with Mayer's hematoxylin and eosin. The immunohistochemistry technique was used to identify cells with signs of proliferation (synthesizing the Ki67 protein), to assess the expression of the synthesis of proteins p53, caspasa 3 and the anti-apoptotic protein bcl-2. The surfaces of titanium structures were studied by scanning electron microscopy. The results were processed statistically. Results. The study investigated the effect of miniplates and miniscrews made of nanostructured and standard titanium on rabbits. The data obtained revealed the nature of the proliferative and apoptotic potentials of osteoblasts in the regenerate, and also indicated an increase in the ratio of the number of cambial cells and cells with an apoptotic dominant in the zone contacting with the nanostructured surface of the titanium screw compared to the titanium screw manufactured by Konmet LLC. Taken together, the obtained data indicate that the use of nanostructured titanium Grade 4 for the production of miniplates and miniscrews has an optimizing effect on reparative osteogenesis. The mechanism of this effect is determined by the presence of osseointegration properties of titanium. Conclusion. Nanostructured titanium is advisable to use for the production of implants, reconstructive titanium plates and other products for bone grafting.