Silver Impregnation Research Articles

Efficacy of silver-impregnated fabric in preventing spread of infection in the wrestling environment.

The best practice for cleaning wrestling mats is using a residual disinfectant with continued antibacterial action. Recently available wash-in silver additives claim to confer a residual effect to fabric. To test the efficacy of laundering with a wash-in silver additive in reducing exposure of athletes to potentially infectious microbes on apparel. 4-part Controlled Laboratory Study/Parallel Group Comparison Study: (1) To test whether fabrics in athletic clothing would be affected differently, we applied bacteria to control fabrics washed in detergent alone and test counterparts washed in detergent plus wash-in silver additive. Bacteria were applied to fabrics, extracted, plated, incubated, and counted. (2) To see if wash-in silver affected various bacteria differently, we washed cotton t-shirts with detergent alone or with detergent plus wash-in silver. We applied four bacterial species commonly found in the wrestling environment. Bacteria were extracted, plated, incubated, and counted. (3) To see if wash-in silver was effective in reducing bacterial contamination during practice, 32 collegiate wrestlers paired off with one wearing a test silver-treated t-shirt, and their partner wearing a control shirt. Wrestler rotations exposed shirts to 2, 4 or 8 wrestlers. Identical swatches of fabric were cut from the t-shirts. Bacteria were extracted, plated, incubated, and counted. (4) We simulated prolonged/repeated bacterial exposure as occurs during tournaments by applying bacteria directly to silver-treated and untreated singlet material repeatedly over time. Test samples were taken at regular intervals to see if bacterial growth was inhibited by the presence of the silver nanoparticles. Bacteria were extracted, plated, incubated, and counted. Laboratory and practice. Collegiate D3 Wrestling Team. Wash-in silver would be considered effective if statistically significant reduction in bacterial count was observed at 95% confidence. Wash-in silver reduced bacterial growth at low levels of contamination but did not significantly reduce bacterial growth at levels seen during contact sport competitions. This was true for all bacterial species and all fabrics tested. The environmental and potential health risks in using a wash-in silver nanoparticle laundry additive in the wash cycle for clothing worn by wrestlers outweigh any potential infection control benefits to these athletes. We do not currently recommend adopting wash-in silver treatment as part of the laundering regimen for wrestling programs until further testing of alternate methods of silver impregnation into sports fabrics has been investigated.

Short-term hyperoxia-induced functional and morphological changes in rat hippocampus.

Excess oxygen (O2) levels may have a stimulating effect, but in the long term, and at high concentrations of O2, it is harmful to the nervous system. The hippocampus is very sensitive to pathophysiological changes and altered O2 concentrations can interfere with hippocampus-dependent learning and memory functions. In this study, we investigated the hyperoxia-induced changes in the rat hippocampus to evaluate the short-term effect of mild and severe hyperoxia. Wistar male rats were randomly divided into control (21% O2), mild hyperoxia (30% O2), and severe hyperoxia groups (100% O2). The O2 exposure lasted for 60 min. Multi-channel silicon probes were used to study network oscillations and firing properties of hippocampal putative inhibitory and excitatory neurons. Neural damage was assessed using the Gallyas silver impregnation method. Mild hyperoxia (30% O2) led to the formation of moderate numbers of silver-impregnated "dark" neurons in the hippocampus. On the other hand, exposure to 100% O2 was associated with a significant increase in the number of "dark" neurons located mostly in the hilus. The peak frequency of the delta oscillation decreased significantly in both mild and severe hyperoxia in urethane anesthetized rats. Compared to normoxia, the firing activity of pyramidal neurons under hyperoxia increased while it was more heterogeneous in putative interneurons in the cornu ammonis area 1 (CA1) and area 3 (CA3). These results indicate that short-term hyperoxia can change the firing properties of hippocampal neurons and network oscillations and damage neurons. Therefore, the use of elevated O2 concentration inhalation in hospitals (i.e., COVID treatment and surgery) and in various non-medical scenarios (i.e., airplane emergency O2 masks, fire-fighters, and high altitude trekkers) must be used with extreme caution.

Standardization of a silver stain to reveal mesoscale myelin in histological preparations of the mammalian brain

BackgroundThe brain is built of neurons supported by myelin, a fatty substance that improves cellular communication. Noninvasive magnetic resonance imaging (MRI) is now able to measure brain structure like myelin and requires histological validation. New methodHere we present work in small and large biomedical model mammals to standardize a silver impregnation method as a high-throughput histological myelin visualization procedure. Specifically, we built a new staining well plate to increase batch size, and then systematically varied the staining and clearing cycles to describe the staining response curve across taxa and conditions. We compared tissues fixed by immersion or perfusion, mounted versus free-floating, and cut as thicker or thinner slices, with two-weeks of post-fixation. ResultsThe staining response curves show optimal staining with a single exposure across taxa when incubation and clearing epochs are held to within 3–9 min. We show that clearing was slower in mounted vs free-floating tissue, and that staining was faster and caused fracturing earlier in thinner sliced and smaller volumes of tissue. Comparison with existing methodsWe developed a batch processing approach to increase throughput while ensuring reproducibility and demonstrate the optimal conditions for fine myelinated fiber morphology visualization with short cycles (<9 minutes). ConclusionsWe present our optimized protocol to reveal mesoscale neuroanatomical myelin content in histology across mammals. This standard staining procedure will facilitate multiscale analyses of myelin content across development as well as in the presence of injury or disease.

Evaluation of schistosoma mansoni nervous system using confocal laser electron microscopy: nerve sensilla and FMRFamide while referring to F-actin abundance

ABSTRACT The chaetotaxy distribution pattern of surface nerve sensilla on the surface of both Schistosoma mansoni and S. haematobium was studied after exposure to silver nitrate treatment followed by light. This distribution pattern could not be achieved earlier due to namelessness, crowded nerve sensilla and twist of most parasites. In S. mansoni, silver impregnation technique displays nerves as ventral nerve cord, ventral sucker nerve that branches to three rings which innervate the ventral sucker. This technique shows the oral sucker ring, ring commissures and bipolar or multipolar nerve cells detected dorsally. In S. haematbium, nerves stained in S. mansoni were also stained in addition to the oral nerve and oral sucker ring. Furthermore, a number of crucial nerves were identified and their innervations to relevant organs were thoroughly examined. A confocal anti-FMRF amide technique was applied to whole-mount worms of both species. In addition to the general tegumental muscles stated previously and stated in this research specific nerves were also described and referred to a specific organ that assumed to innervate. In conclusion, the findings of the current study differentiate between S. mansoni and S. mansoni using chaetotaxy map as a step for spotlighting on FMRF amidergic components as antihelmintic target in future.

Dynamics of structural transformations of intervertebral discs in humans in the fetal period

BACKGROUND:The process of formation of the intervertebral disc structure remains poorly understood. Therefore, evaluating the forecasts for the development of the disc’s fibrous component as the foundation of its strength and elastic properties in the postnatal period seems appropriate.&#x0D; AIM:To identify microstructural transformations of the annulus fibrosus and nucleus pulposus in the fetal period and compare them with the ontogenetic orientation of the spinal motion segment.&#x0D; MATERIAL AND METHODS:The study material included 150intervertebral discs obtained during autopsy of 50fetuses. The gestational age of 42fetuses was in the early fetal period, 8fetuses were in the late fetal period. Intervertebral discs СV–СVI, ThV–ThVI, LV–SIwere examined in each fetus. All histological preparations were stained with hematoxylin and eosin, silver impregnation method, PAS reaction, Alcian blue (pH=1.0), Van Gieson and Weigert stains. For intergroup comparisons, the Kruskal–Wallis, Mann–Whitney, and Newman–Keuls tests were used. Differences were considered statistically significant atp≤0.05.&#x0D; RESULTS:Elastic fibers in the late fetal period were found in the annulus fibrosus and the peripheral zone of the nucleus pulposus. The intensity of staining of collagen and elastic fibers was more pronounced in the outer layers of the annulus fibrosus. Analysis of the parameters of the vascular-connective tissue formations showed an increase in their number (p=0.0368) and an increase in the number of vessels in the vascular-connective tissue formations of the intervertebral disc (p=0.0449) in the direction of the lower levels of the spinal motion segments. Differences in these two indicators were obtained between the СV–СVIand LV–SIdiscs, as well as between the СV–СVIand ThV–ThVIdiscs. In relation to all the studied morphometric parameters of the vascular-connective tissue formations in the cervical and thoracic spine, no statistically significant differences were obtained between the early and late fetal periods.&#x0D; CONCLUSION:The development of the intervertebral disc occurs from the peripheral parts to the center; the source for its further development is the fibrous ring.

Evolution of staining methods in neuroanatomy: Impetus for emanation of neuron doctrine during the turn of 20th century.

The nervous system is distinctive as compared to other tissue systems in human body owing to intricate structural organization. Histological studies played a key role in unveiling complex details of nervous tissue. However, the process of developing suitable staining method for nerve cells was arduous and spanned across almost half a century. The present study explored details of the journey involving quest for propitious staining method in neuroanatomy culminating in promulgation of neuron doctrine at the onset of 20th century. Initial efforts involving hematoxylin (including its diverse modifications) and subsequent adoption of analogous dye-based stains (like Nissl's method) had limited success in visualization of different parts of a nerve cell and structural details of nervous tissue. This was due to inability of dye-based stains to penetrate the connective tissue sheath of nervous tissue. Eventually, advent of metallic stains in form of silver impregnation method (Golgi stain), reduced silver impregnation method with gold stain (Cajal's stain) and silver carbonate staining method of Río-Hortega unraveled the structure of nervous tissue. The evolution of staining methods catalyzed the refinement of theories pertinent to constitution of nervous tissue. Golgi's staining led to emergence of reticular theory (neurons exist as a network) and Nissl's staining was the basis of the concept of Nervösen Grau (nerve cells and glial cells are embedded in mass of gray matter). Finally, Cajal's staining method successfully elucidated the complex anatomy of nerve terminals and resulted in emanation of neuron doctrine (neurons exists as individual units with adjacent connections).

Cytological, Histopathological and Immunohistochemical Studies on Naturally Occurring Cutaneous Melanomas in Cattle in and Around Namakkal, Tamil Nadu, India

Background: Melanomas are tumours that originate from melanoblasts. The studies on bovine tumours are of clinical concern since some melanocytic tumours are congenital and occur in young animals. The present work was carried out to study the pathomorphological characteristics of melanomas in cattle for early diagnosis and intervention. Methods: Tumour samples were collected from suspected cases of neoplasia from four animals during the study period. Data pertaining to the animals were collected. Fine needle aspiration cytology and impression smear were stained with Giemsa stain. Tissue samples were collected in 10 per cent neutral buffered formalin for histopathological examination. Special staining with Fontana silver stain and immunohistochemistry with Melan- A and S- 100 markers for confirmatory diagnosis of melanin pigment was done. Result: Cytology revealed neoplastic cells with vacuolation and pleomorphism. Cytoplasm contained abundant melanin pigments. Microscopically, the cytoplasm contained abundant brown, black coloured melanin pigment granules. Tissue sections stained with Fontana silver impregnation method revealed the presence of black coloured granules. Immunohistochemistry with Melan-A and S-100 revealed strong expression of brown coloured reaction in the cytoplasm of the neoplastic cells.

Topographical anatomy of the subthalamic region with special interest in the human medial forebrain bundle.

The medial forebrain bundle (MFB) is a novel promising deep brain stimulation (DBS) target in severe affective disorders that courses through the subthalamic region according to tractography studies. Its potential therapeutic role arose in connection with the development of hypomania during stimulation of the subthalamic nucleus (STN) in Parkinson's disease, offering an alternative explanation for the occurrence of this side effect. However, until now its course exclusively described by tractography had not yet been confirmed by any anatomical method. The aim of this study was to fill this gap as well as to provide a detailed description of the fiber tracts surrounding the STN to facilitate a better understanding of the background of side effects occurring during STN DBS. Ten human cadaveric brains (20 hemispheres) and 100 healthy subjects (200 hemispheres) from the S500 Release of the Human Connectome Project were involved in this study. Nineteen hemispheres were dissected according to Klingler's method. One additional hemisphere was prepared for histological examinations to validate the macroscopical results and stained with neurofibril silver impregnation according to Krutsay. The authors also aimed to reconstruct the MFB using tractography and correlated the results with their dissections and histological findings. The white matter connections coursing through the subthalamic region were successfully dissected. The ansa lenticularis, lenticular fasciculus, thalamic fasciculus, ipsi- and contralateral cerebellar fibers, and medial lemniscus were revealed as closely related fiber tracts to the STN. However, the existence of a distinct fiber bundle corresponding to the MFB described by tractography could not be identified. Using tractography, the authors showed that the depiction of the streamlines representing the MFB was also strongly dependent on the threshold parameters. According to this study's findings, the streamlines of the MFB described by tractography arise from the limitations of the diffusion-weighted MRI fiber tracking method and actually correspond to subthalamic fiber bundles, especially the ansa lenticularis and lenticular fasciculus, which erroneously continue in the anterior limb of the internal capsule, toward the prefrontal cortex.

Karyological Study of Acanthocephalus lucii (Echinorhynchida): The Occurrence of B Chromosomes in Populations from PCB-Polluted Waters

In this study, we performed a cytogenetic analysis of Acanthocephalus lucii specimens from three sites with different levels of environmental pollution. Standard and fluorochrome staining (CMA3/DAPI), fluorescence in situ hybridization (FISH) with 18S rDNA and histone H3 probes, and silver impregnation were performed. Chromosome complements of 2n = 7/8 (male/female), n = 1m + 2sm + 1a (X), and CMA3-positive bands in all chromosomes were found in all three populations. FISH revealed one 18S rDNA locus on the X chromosome and one locus of H3 histone genes on the first chromosome pair. At the intraspecific level, the populations differed in the presence of supernumerary B chromosomes, which were found in all specimens from Zemplínska Šírava and in 89.4% of specimens from the Laborec River, but not at the reference site. The first two sites are considered to be water bodies with high toxin contamination. Based on this fact, we assume an increased frequency of chromosome breaks leading to the formation of DNA fragments that have the potential to form B chromosomes. The present results add to the very limited data on the organization of multigene families in the genome of Acanthocephala and suggest a possible causal link between water pollution and the occurrence of B chromosomes in fish parasites.

Role of Angiogenesis and Endothelialmesenchymal Transition in Bone Marrow Fibrosis Associated with Haematopoietic Neoplasms: A Cross-sectional Immunohistochemical Analysis

Introduction: The bone marrow examination is an essential investigation in the diagnosis and management of many haematological disorders. The integration of all investigations, including peripheral blood analysis, bone marrow aspirate, and trephine biopsy findings, along with supplementary tests such as immunophenotyping, cytogenetic analysis, and molecular genetic studies, is crucial for arriving at a final diagnosis. Aim: To assess the presence of reticulin fibres in bone marrow biopsy sections in haematological malignancies, to evaluate the grade of BMF associated with haematological malignancies and to assess the role of angiogenesis using IHC markers in various haematological malignancies. Materials and Methods: A cross-sectional study was conducted in the Department of Pathology at the National Institute of Pathology, Safdarjung Hospital, New Delhi, India in 2009 for a duration of 18 months. Thirty-eight patients with a diagnosis of Acute Myeloid Leukaemia (AML), Acute Lymphoblastic Leukaemia (ALL), Chronic Myeloid Leukaemia (CML), and Chronic Lymphoproliferative Disorder (CLPD) were studied. Bone marrow biopsies were taken, fixed in 10% formalin, and decalcified in 10% Ethylene Diamine Tetraacetic Acid (EDTA). Routine paraffin embedding was performed, and serial sections of 4 µm were obtained on poly-L-lysine-coated slides for Immunohistochemistry (IHC) {Vimentin, Vascular Endothelial Growth Factor (VEGF), CD-34, Smooth Muscle Actin (SMA)}. The presence of reticulin fibres in the bone marrow biopsy sections was assessed using two special stains: Gomori’s Silver Impregnation and Masson’s Trichrome. Fibrosis was quantified according to the Baurmeister 0-4 grading system of Bone Marrow Fibrosis (BMF). Results: The results of the present study suggest that Endothelial to Mesenchymal Transition (EndMT) may play a role in the pathogenesis of BMF. Various grades of fibrosis were observed, with 15 cases (39.47%) in Grade 3, followed by 11 cases (28.95%) in Grade 2, 8 cases (21.05%) in Grade 1, and 4 cases (10.53%) in Grade 4. Conclusion: BMF was a significant finding even in the early stages of the majority of the lesions studied and was closely linked with angiogenesis. This study showed that angiogenesis plays an important role in the pathogenesis of haematological neoplasms and that VEGF is a prominent stimulus in the majority of these disorders. Additionally, this study suggests that EndMT has a possible role in the pathogenesis of BMF.Introduction: The bone marrow examination is an essential investigation in the diagnosis and management of many haematological disorders. The integration of all investigations, including peripheral blood analysis, bone marrow aspirate, and trephine biopsy findings, along with supplementary tests such as immunophenotyping, cytogenetic analysis, and molecular genetic studies, is crucial for arriving at a final diagnosis. Aim: To assess the presence of reticulin fibres in bone marrow biopsy sections in haematological malignancies, to evaluate the grade of BMF associated with haematological malignancies and to assess the role of angiogenesis using IHC markers in various haematological malignancies. Materials and Methods: A cross-sectional study was conducted in the Department of Pathology at the National Institute of Pathology, Safdarjung Hospital, New Delhi, India in 2009 for a duration of 18 months. Thirty-eight patients with a diagnosis of Acute Myeloid Leukaemia (AML), Acute Lymphoblastic Leukaemia (ALL), Chronic Myeloid Leukaemia (CML), and Chronic Lymphoproliferative Disorder (CLPD) were studied. Bone marrow biopsies were taken, fixed in 10% formalin, and decalcified in 10% Ethylene Diamine Tetraacetic Acid (EDTA). Routine paraffin embedding was performed, and serial sections of 4 µm were obtained on poly-L-lysine-coated slides for Immunohistochemistry (IHC) {Vimentin, Vascular Endothelial Growth Factor (VEGF), CD-34, Smooth Muscle Actin (SMA)}. The presence of reticulin fibres in the bone marrow biopsy sections was assessed using two special stains: Gomori’s Silver Impregnation and Masson’s Trichrome. Fibrosis was quantified according to the Baurmeister 0-4 grading system of Bone Marrow Fibrosis (BMF). Results: The results of the present study suggest that Endothelial to Mesenchymal Transition (EndMT) may play a role in the pathogenesis of BMF. Various grades of fibrosis were observed, with 15 cases (39.47%) in Grade 3, followed by 11 cases (28.95%) in Grade 2, 8 cases (21.05%) in Grade 1, and 4 cases (10.53%) in Grade 4. Conclusion: BMF was a significant finding even in the early stages of the majority of the lesions studied and was closely linked with angiogenesis. This study showed that angiogenesis plays an important role in the pathogenesis of haematological neoplasms and that VEGF is a prominent stimulus in the majority of these disorders. Additionally, this study suggests that EndMT has a possible role in the pathogenesis of BMF.

Longitudinal bond strength of a universal adhesive and chemical dentin characterization under different acid etching protocols.

This study aimed to analyze the longitudinal bond strength of a universal adhesive and chemically characterize the dentin substrate under different acid etching protocols. Dentin samples were etched with polyacrylic acid 25% (PAA) for 10 seconds (n=3) and phosphoric acid 32% (PA) for 15 seconds (n=3) and analyzed by Fourier transform infrared spectroscopy - attenuated total reflectance (FTIR-ATR) before and after treatment. For collagen degradation, samples (n=12) were divided into 3 groups: PAA, PA, and Deionized water (control), and analyzed by the quantity of solubilized type I collagen C-terminal cross-linked telopeptides and solubilized C-terminal peptide in relation to total protein concentration (ICTPtp and CTXtp) and by their ultimate tensile strength (UTS). For the adhesive interface analysis, dentin samples (n=72) were divided into 3 groups: PAA, PA, and Self-etch (SE), and subdivided into 2 groups: 24 h (baseline) and 1 year. The following tests were performed: microtensile bond strength (μTBS) (n=48), scanning electron microscopy (SEM) (n=12), and nanoleakage (n=12). The FTIR of PAA showed lower reduction of the peaks in the phosphate group when compared to PA. For ICTPtp, PA showed a significantly higher value. For CTXtp, PA and PAA groups failed to statically differ from each other. UTS was significantly lower for PA. For μTBS, storage time significantly affected bond strength. The results were unaffected by the etching protocol. For SEM, after 1 year, PA had little evidence of degradation in the upper third of the adhesive interface in comparison to the other groups. Nanoleakage showed no considerable silver impregnation after 1 year in the SE group. The use of PAA prior to a universal adhesive (when compared to PA) represents a less aggressive type of etching to dentin. However, self-etching still seems to be the best option for universal adhesive systems that have functional monomers in their composition.

Pitiose esofágica e gástrica em cão

ABSTRACT: Pythiosis is the disease caused by aquatic oomycetes of the genus Pythium. In dogs the typical course of the disease involves the gastrointestinal tract, characterized by transmural thickening of the stomach or intestines. However, infection by Pythium spp. has only rarely been recognized as a cause of esophagitis in dogs. Thus, the present reports a case of esophageal and gastric pythiosis in an eight-month-old female pit bull dog. The dog was attended at the hospital after two months presenting regurgitation and dyspnea. It was reported that the dog lived in an urban area and had no previous sanitary issues. At clinical examination it was noted that the dog presented crackling sounds at pulmonary auscultation. A support therapy accompanied by antibiotics has been employed; however, 22 days after hospitalization the clinical condition worsened, and the dog died. At necropsy, the wall of the distal segment of the esophagus and the cardia and part of the fundus of the stomach were expanded by a focal extensive irregular intramural annular mass. Additionally, there was a transmural esophageal fistula. At histology, the walls of the esophagus and stomach were extensively expanded by multifocal extensive areas of necrosis, associated with a pyogranulomatous infiltrate and abundant granulation tissue containing multiple negative images of hyphae that were highlighted by silver impregnation (Grocott). Furthermore, immunohistochemistry and PCR for P. insidiosum were both positive in samples of paraffin-embedded esophageal tissue.

Application of Several Special Staining Methods for Paraffin Sections on Epon-Embedded Semithin Sections

Aim: This study aimed to compare several specific staining protocols recommended for paraffin sections and toluidine blue and light green double staining combination to be tried for the first time with routine toluidine blue staining on semithin epon sections.&#x0D; Material and Methods: Samples of 1x1x1 mm were taken from the liver, skin, and aorta tissues of Wistar albino adult rats. Tissue samples were fixed with 5% glutaraldehyde at +4º C overnight, postfixed with 1% osmium tetroxide for one hour, and then, blocked with Epon 812 after processing. Semithin sections of 1 μm thickness were obtained from the epon blocks. Sections were stained with Altmann’s method (for mitochondria), Verhoeff’s method (for elastic fibers), Gordon&amp;Sweets’ silver impregnation method (for type III collagen), toluidine blue and light green double staining combination (for type I collagen) and routine toluidine blue method.&#x0D; Results: In liver sections, mitochondria in hepatocytes were differentiated by the Altmann method, and stromal type III collagen fibers were distinguished with Gordon&amp;Sweets’ method. Elastic lamellar structures were easily observed in black in the aortic sections stained with the Verhoeff method. Successful results were obtained in the staining of dermal type I collagen with toluidine blue and light green double staining in skin sections.&#x0D; Conclusion: Since the specific staining tried for the first time gave positive results in epon sections, it was concluded that these methods can be used to determine the localization of cellular and intercellular components that are aimed to be examined at the ultrastructural level.

Treating Periprosthetic Joint Infection With Silver-Impregnated Carbon-Coated Spacers: Mid-Term Outcomes

Background. Periprosthetic joint infection (PJI) is a severe complication of arthroplasty. The widely accepted treatment standard for PJI is a two-stage revision arthroplasty involving the articulating spacers. The implant surface provides an ideal environment for bacterial adhesion, facilitating mature biofilm formation. To prevent bacterial adhesion effectively, the surface of the implanted device must be modified with an efficient coating. The ability of a modified coating based on two-dimensional linear carbon chains (2D LCC) with silver (Ag) impregnation to inhibit biofilm formation and provide efficient bacterial eradication has been investigated in several experimental studies. However, there is a lack of publications on clinical studies evaluating the effectiveness of such coatings.&#x0D; The aim of the study — to assess mid-term outcomes of knee and hip PJI treatment using spacers coated with two-dimensional linear carbon chains impregnated with silver.&#x0D; Methods. This study is based on the results of the examination and two-stage revision arthroplasty of 144 patients with newly diagnosed knee and hip PJI. Patients were divided into two groups: the first (main) group received articulating spacers coated with 2D LCC+Ag, while the second (control) group received articulating spacers with antibiotics. Anamnestic, clinical, laboratory, microbiological, and statistical methods were used in this study. The evaluation of short-term results was performed using the KSS, Harris, VAS, and EQ-5D-5L scales at 3 months after surgery, and mid-term results were assessed at 2 years.&#x0D; Results. The study confirmed the high antibiofilm activity and safety of spacers coated with 2D LCC+Ag. Both groups showed a reduction in inflammation markers during treatment. Before the second stage of treatment, both groups experienced a statistically significant decrease in CRP, procalcitonin, and presepsin levels, as well as synovial cytosis and neutrophil content. The frequency of recurrences after two-stage treatment was significantly lower in the first group compared to the second group. In the mid-term period, the first group had higher scores on the KSS and Harris scales by 20.5 and 7.0 points, respectively. Results on the EQ-5D-5L were 10/0.08 points higher, and the intensity of pain according to VAS was three times lower in the first group.&#x0D; Conclusion. The use of spacers coated with 2D LCC+Ag allows for a faster resolution of the inflammatory process, reduces the incidence of PJI recurrences, and predicts active protection of the implant surface from microbial colonization and biofilm formation. This, combined with antibiotic prophylaxis, provides a favorable therapeutic and preventive effect against PJI recurrence.

Improving HCC Prognostic Models after Liver Resection by AI-Extracted Tissue Fiber Framework Analytics.

Despite advances in diagnostic and treatment technologies, predicting outcomes of patients with hepatocellular carcinoma (HCC) remains a challenge. Prognostic models are further obscured by the variable impact of the tumor properties and the remaining liver parenchyma, often affected by cirrhosis or non-alcoholic fatty liver disease that tend to precede HCC. This study investigated the prognostic value of reticulin and collagen microarchitecture in liver resection samples. We analyzed 105 scanned tissue sections that were stained using a Gordon and Sweet's silver impregnation protocol combined with Picric Acid-Sirius Red. A convolutional neural network was utilized to segment the red-staining collagen and black linear reticulin strands, generating a detailed map of the fiber structure within the HCC and adjacent liver tissue. Subsequent hexagonal grid subsampling coupled with automated epithelial edge detection and computational fiber morphometry provided the foundation for region-specific tissue analysis. Two penalized Cox regression models using LASSO achieved a concordance index (C-index) greater than 0.7. These models incorporated variables such as patient age, tumor multifocality, and fiber-derived features from the epithelial edge in both the tumor and liver compartments. The prognostic value at the tumor edge was derived from the reticulin structure, while collagen characteristics were significant at the epithelial edge of peritumoral liver. The prognostic performance of these models was superior to models solely reliant on conventional clinicopathologic parameters, highlighting the utility of AI-extracted microarchitectural features for the management of HCC.

Morphological peculiarities of revascularization of perifocal areas of brain infarction

Aim. To study pathomorphological signs of revascularization of perifocal areas of brain infarction in the dynamics of the acute period of the disease. Material and methods. Perifocal areas of brain infarction in patients who died at the 1st, 3rd, 7th, and 14th days were studied. General histopathological study was followed by 10 % ammonium silver impregnation and immunohistochemical examination using monoclonal antibodies Mo a-Hu CD34, Clone QBEnd/10 (“Thermo Fisher Scientific Inc.” – USA) and Mo a-Hu CD105 Endoglin were used. clone: SN6h1 (“DAKO”, Denmark). Vascular density and total cross-sectional area of vessels were studied. For the morphometric study, Videotest – Morphology 5.2.0.158 software (VideoTest LLC) was used. Statistical processing of the obtained data was carried out using the Statistica® for Windows 13.0 software. Results. According to the data of CD34 expression, from the 3rd day in the perifocal areas of brain infarction, the angiogenesis processes were increased: the density of blood vessels increased by 18.48 %, and the total cross-sectional area of vessels increased by 1.93 times. On the 7th day, the maximal vessel density and total cross-sectional area of vessels were registered in these areas; there was a vascular budding, formation of endothelial proliferates, and increased expression of CD105 on the vascular endothelium and pericytes, which indicates the active participation of the latter in angiogenesis. On the 14th day, blood vessel density and indicators of the total cross-sectional area of vessels decreased. During this period, the density of the astrocytes increased in the infarct’s perifocal areas with hypertrophy of their bodies and processes. Conclusions. According to CD34 and CD105 expression data, in the perifocal areas of cerebral infarction, the processes of angiogenesis gradually increase from the 3rd day manifesting by increase in the blood vessel density and total vascular cross-sectional area, as well as in the vascular budding and the formation of endothelial proliferates. Vascular remodeling is accompanied by increased density and hypertrophy of astroglia in the perifocal areas of cerebral infarction.

Histological and cytochemical analysis of the brain under conditions of hypobaric hypoxia-induced oxygen deficiency in albino rats

High altitude sickness is a life-threatening disease that occurs among acclimatized individuals working or living at a high altitude accompanied by hypobaric hypoxia exposure. The prolonged influence of hypobaric hypoxia on the brain may trigger neuronal damage and cell death due to an oxygen deficiency. The purpose of the current study was to investigate the histomorphological changes in the hippocampus, cerebral cortex, cerebellar cortex, and striatum of the rat’s brain following chronic hypobaric hypoxia. Fourteen albino rats were used for this investigation. The animals were exposed to chronic hypobaric hypoxia in the special decompression chamber at an altitude of 7000 m for 7 days. The histological analysis was conducted via toluidine staining and silver impregnation. DNA damage and cell apoptosis were assessed via Feulgen staining. The histochemical assessment revealed increased dark neurons in the hippocampus with cell swelling. Silver impregnation showed increased argyrophilic neurons in the cerebellar cortex, striatum, CA1 subfield of the hippocampus, and cerebral cortex. The cytochemical analysis determined the increased apoptotic cells with hyperchromatic condensation and pyknosis in the hippocampus subfields and cerebral cortex. In addition, it has been observed that hypoxia has resulted in small hemorrhages and perivascular edema within the cerebellar and cerebral cortex.The results indicate brain injury observed in the various parts of the brain towards hypobaric hypoxia, however, the hippocampus showed greater vulnerability against hypoxic exposure in comparison to the striatum, cerebellum, and cerebral cortex. These changes support our insights regarding brain intolerance under conditions of hypoxia-induced oxygen deficiency and its histomorphological manifestations.

Comparative neuroanatomy of the central nervous system in web-building and cursorial hunting spiders.

Spiders (Araneae) include cursorial species that stalk their prey and more stationary species that use webs for prey capture. While many cursorial hunting spiders rely on visual cues, web-building spiders use vibratory cues (mechanosensation) for prey capture. We predicted that the differences in primary sensory input between the species are mirrored by differences in the morphology/architecture of the central nervous system (CNS). Here, we investigated the CNS anatomy of four spider species, two cursorial hunters Pardosa amentata (Lycosidae) and Marpissa muscosa (Salticidae), and two web-building hunters Argiope bruennichi (Araneidae) and Parasteatoda tepidariorum (Theridiidae). Their CNS was analyzed using Bodian silver impregnations, immunohistochemistry, and microCT analysis. We found that there are major differences between species in the secondary eye pathway of the brain that pertain to first-order, second-order, and higher order brain centers (mushroom bodies [MB]). While P. amentata and M. muscosa have prominent visual neuropils and MB, these are much reduced in the two web-building species. Argiope bruennichi lacks second-order visual neuropils but has specialized photoreceptors that project into two distinct visual neuropils, and P. tepidariorum lacks MB, suggesting that motion vision might be absent in this species. Interestingly, the differences in the ventral nerve cord are much less pronounced, but the web-building spiders have proportionally larger leg neuropils than the cursorial spiders. Our findings suggest that the importance of visual information is much reduced in web-building spiders, compared to cursorial spiders, while processing of mechanosensory information requires the same major circuits in both web-building and cursorial hunting spiders.

Mild hypoxia-induced structural and functional changes of the hippocampal network.

Hypoxia causes structural and functional changes in several brain regions, including the oxygen-concentration-sensitive hippocampus. We investigated the consequences of mild short-term hypoxia on rat hippocampus in vivo. The hypoxic group was treated with 16% O2 for 1 h, and the control group with 21% O2. Using a combination of Gallyas silver impregnation histochemistry revealing damaged neurons and interneuron-specific immunohistochemistry, we found that somatostatin-expressing inhibitory neurons in the hilus were injured. We used 32-channel silicon probe arrays to record network oscillations and unit activity from the hippocampal layers under anaesthesia. There were no changes in the frequency power of slow, theta, beta, or gamma bands, but we found a significant increase in the frequency of slow oscillation (2.1-2.2 Hz) at 16% O2 compared to 21% O2. In the hilus region, the firing frequency of unidentified interneurons decreased. In the CA3 region, the firing frequency of some unidentified interneurons decreased while the activity of other interneurons increased. The activity of pyramidal cells increased both in the CA1 and CA3 regions. In addition, the regularity of CA1, CA3 pyramidal cells' and CA3 type II and hilar interneuron activity has significantly changed in hypoxic conditions. In summary, a low O2 environment caused profound changes in the state of hippocampal excitatory and inhibitory neurons and network activity, indicating potential changes in information processing caused by mild short-term hypoxia.

Staphylococcus aureus Adhesion and Biofilm Formation on Vascular Polyester Grafts are Inhibited In Vitro by Triclosan

This study evaluated Staphylococcus aureus adhesion and biofilm formation on vascular grafts, which has seldom been investigated. Adhesion and biofilm formation capabilities of three methicillin susceptible S.aureus strains (one biofilm forming reference strain and two clinical isolates) on five different vascular biomaterials were evaluated invitro, including polyester (P), P+ gelatin (PG), P+ collagen (PC), PC+ silver (PCS), and PCS+ triclosan (PCST). Staphylococcus aureus adhesion on grafts was evaluated after one hour of culture and biofilm formation after 24 hours of culture by four different methods: spectrophotometry after crystal violet staining; sonicate fluid culture; metabolic assay; and scanning electron microscopy (SEM). Optical density was compared using Mann-Whitney pairwise test, and bacterial counts using Wilcoxon pairwise test. PCST grafts were most efficient in preventing S.aureus adhesion and biofilm formation, regardless of the method used. Bacterial counts and metabolic activity were significantly lower on PCST grafts after 24 hours (5.65 vs. 9.24 [PCS], 8.99 [PC], 8.82 [PG], and 10.44 log10 CFU/mL [P]; p < .015), and only PCST grafts were bactericidal. Biofilm formation was significantly diminished on PCST grafts compared with all other grafts (p < .001). Bacterial viability and metabolic activity after 24 hours were more impaired on PG compared with PC graft, and were surprisingly higher on PCS compared with PC grafts. Biofilm biomass formed after exposure to P, PG, PC, and PCS grafts was also reduced after 24 hours of incubation with PCST grafts (p < .001). After 24 hours, few bacteria were visible by SEM on PCST grafts, whereas bacterial biofilm colonies were clearly identified on other graft surfaces. Triclosan impregnated PCST grafts appeared to interfere with S.aureus adhesion from early stages of biofilm formation invitro. Silver impregnation was not efficient in preventing biofilm formation, and collagen coating promoted S.aureus biofilm formation more than gelatin coating.