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Drug Delivery and Translational Research Mar 2023This study is aimed to fabricate tetanus toxoid laden microneedle patches by using a polymeric blend comprising of polyvinyl pyrrolidone and sodium carboxymethyl...
This study is aimed to fabricate tetanus toxoid laden microneedle patches by using a polymeric blend comprising of polyvinyl pyrrolidone and sodium carboxymethyl cellulose as base materials and sorbitol as a plasticizer. The tetanus toxoid was mixed with polymeric blend and patches were prepared by using vacuum micromolding technique. Microneedle patches were evaluated for physical attributes such as uniformity of thickness, folding endurance, and swelling profile. Morphological features were assessed by optical and scanning electron microscopy. In vitro performance of fabricated patches was studied by using bicinchoninic acid assay (BCA). Insertion ability of microstructures was studied in vitro on model skin parafilm and in vivo in albino rat. In vivo immunogenic activity of the formulation was assessed by recording immunoglobulin G (IgG) levels, interferon gamma (IFN-γ) levels, and T-cell (CD4 and CD8) count following the application of dosage forms. Prepared patches, displaying sharp-tipped and smooth-surfaced microstructures, remained intact after 350 ± 36 foldings. Optimized microneedle patch formulation showed ~ 74% swelling and ~ 85.6% vaccine release within an hour. The microneedles successfully pierced parafilm. Histological examination of microneedle-treated rat skin confirmed disruption of epidermis without damaging the underneath vasculature. A significant increase in IgG levels (~ 21%), IFN-γ levels (~ 30%), CD4 (~ 41.5%), and CD8 (~ 48.5%) cell count was observed in tetanus vaccine-loaded microneedle patches treated albino rats with respect to control (untreated) group at 42nd day of immunization. In conclusion, tetanus toxoid-loaded microneedle patches can be considered as an efficient choice for transdermal delivery of vaccine without inducing pain commonly experienced with hypodermic needles.
Topics: Administration, Cutaneous; Drug Delivery Systems; Immunoglobulin G; Needles; Paraffin; Polymers; Tetanus Toxoid; Transdermal Patch; Animals; Rats
PubMed: 36253518
DOI: 10.1007/s13346-022-01249-9 -
Andrology Nov 2022The unique anatomy of the male reproductive organ reflects its complex function from sperm maturation to their storage for months until emission. Since light microscopy...
BACKGROUND
The unique anatomy of the male reproductive organ reflects its complex function from sperm maturation to their storage for months until emission. Since light microscopy in two dimensions (2d) cannot sufficiently demonstrate its complex morphology, a comprehensive visualization is required to identify pathologic alterations in its entire anatomical context.
OBJECTIVES
Aim of this study was to use three-dimensional (3d) light sheet fluorescence microscopy (LSFM) to visualize entire murine testes in 3d, label-free and at subcellular resolution, and to assign local autofluorescence to testicular and deferent structures.
MATERIALS AND METHODS
Murine testes were fixed with four different fixatives and subsequently cleared with benzoic acid/benzyl benzoate. Hereafter, complete murine testes were scanned with LSFM with different fluorescence filter sets and subsequently embedded in paraffin for further conventional planar histology.
RESULTS
Autofluorescence signals of the murine reproductive organ allowed the unambiguous identification of the testicular anatomy from the seminiferous tubules to the vas deferens with their specific stratification independent of the used fixative. Blood vessels were visualized from the pampiniform plexus to the small capillaries of single tubules. Moreover, due to the specific intrinsic fluorescence properties of the efferent ducts and the epididymis, luminal caliber, the epithelial stratification and retronuclear cytoplasmic inclusions gave a unique insight into the interface of both morphological structures. Subsequent 2d histology confirmed the identified morphological structures.
DISCUSSION
LSFM analysis of the murine reproductive organ allows due to its intrinsic fluorescence a simple, label-free 3d assessment of its entire duct morphology, the epithelial composition, and the associated blood supply in its anatomical relation.
CONCLUSION
LSFM provides the technical basis for comprehensive analyses of pathologically altered murine testes in its entirety by depicting specific autofluorescence. Thereby it facilitates mouse studies of testicular disease or their drug-related alterations in more detail potentially for clinical translation assessing human testicular biopsies.
Topics: Animals; Benzoic Acid; Epididymis; Fixatives; Humans; Male; Mice; Microscopy, Fluorescence; Paraffin; Semen; Testis
PubMed: 36082398
DOI: 10.1111/andr.13292 -
Laboratory Investigation; a Journal of... May 2023Tissue structures, phenotypes, and pathology are routinely investigated based on histology. This includes chemically staining the transparent tissue sections to make...
Tissue structures, phenotypes, and pathology are routinely investigated based on histology. This includes chemically staining the transparent tissue sections to make them visible to the human eye. Although chemical staining is fast and routine, it permanently alters the tissue and often consumes hazardous reagents. On the other hand, on using adjacent tissue sections for combined measurements, the cell-wise resolution is lost owing to sections representing different parts of the tissue. Hence, techniques providing visual information of the basic tissue structure enabling additional measurements from the exact same tissue section are required. Here we tested unstained tissue imaging for the development of computational hematoxylin and eosin (HE) staining. We used unsupervised deep learning (CycleGAN) and whole slide images of prostate tissue sections to compare the performance of imaging tissue in paraffin, as deparaffinized in air, and as deparaffinized in mounting medium with section thicknesses varying between 3 and 20 μm. We showed that although thicker sections increase the information content of tissue structures in the images, thinner sections generally perform better in providing information that can be reproduced in virtual staining. According to our results, tissue imaged in paraffin and as deparaffinized provides a good overall representation of the tissue for virtually HE-stained images. Further, using a pix2pix model, we showed that the reproduction of overall tissue histology can be clearly improved with image-to-image translation using supervised learning and pixel-wise ground truth. We also showed that virtual HE staining can be used for various tissues and used with both 20× and 40× imaging magnifications. Although the performance and methods of virtual staining need further development, our study provides evidence of the feasibility of whole slide unstained microscopy as a fast, cheap, and feasible approach to producing virtual staining of tissue histology while sparing the exact same tissue section ready for subsequent utilization with follow-up methods at single-cell resolution.
Topics: Male; Humans; Hematoxylin; Eosine Yellowish-(YS); Paraffin; Microscopy; Staining and Labeling
PubMed: 36801642
DOI: 10.1016/j.labinv.2023.100070 -
Cell and Tissue Research Sep 2021The bone is a dynamic and metabolically active organ in which growth and resorption of the osteochondral matrix is orchestrated by osteoblasts and osteoclasts. For...
The bone is a dynamic and metabolically active organ in which growth and resorption of the osteochondral matrix is orchestrated by osteoblasts and osteoclasts. For decalcified paraffin-embedded specimens, decalcifying agents alter the staining intensity, and excess decalcification interferes with bone staining. Robust bone staining methods independent of the decalcification conditions and animal species are lacking. In this study, we have developed a novel polychrome staining method, named JFRL staining, which stains the components of osteochondral tissue in different colors. With this staining we could visualize the hyaline cartilage as blue by alcian blue, osteoid as red by picrosirius red, and mineralized bone as green by picro-light green SF or picro-naphthol green B and easily distinguished osteoblasts, osteocytes, and osteoclasts. In mineralized bone, this staining revealed the obvious lamellar structures and woven bone. Notably, this staining was independent of the decalcification conditions and experimental animal species examined. To verify the usefulness of JFRL staining, we observed cotton rat tail which has shorter length and shows a false autotomy. The caudal vertebrae were normally developed via endochondral ossification without a fracture plane. At 6 months of age, the number of chondrocytes declined and the hypertrophic zone was absent at the epiphyseal plate, which might reflect the shorter tail. In conclusion, JFRL staining is the first method to simultaneously distinguish osteochondral matrix and bone cells in one section regardless of decalcifying conditions. This robust staining will provide new information for a wide number of biomedical fields, including bone development, physiology, and pathology.
Topics: Animals; Bone Development; Male; Mice; Osteochondritis; Paraffin
PubMed: 34410480
DOI: 10.1007/s00441-021-03516-6 -
Chemosphere Mar 2022Technical chlorinated paraffins (CPs) are produced via radical chlorination of n-alkane feedstocks with different carbon chain-lengths (∼C-C). Short-chain CPs (SCCPs,...
Technical chlorinated paraffins (CPs) are produced via radical chlorination of n-alkane feedstocks with different carbon chain-lengths (∼C-C). Short-chain CPs (SCCPs, C-C) are classified as persistent organic pollutants (POPs) under the Stockholm Convention. This regulation has induced a shift to use longer-chain CPs as substitutes. Consequently, medium-chain (MCCPs, C-C) and long-chain (LCCPs, C) CPs have become dominant homologues in recent environmental samples. However, no suitable LCCP-standard materials are available. Herein, we report on the chemical synthesis of single-chain C-CP-materials, starting with a pure n-alkane and sulfuryl chloride (SOCl). Fractionation of the crude product by normal-phase liquid-chromatography and pooling of suitable fractions yielded in four C-CP-materials with different chlorination degrees (m = 39-52%). In addition, polar side-products, tentatively identified as sulfite-, sulfate- and bis-sulfate-diesters, were separated from CPs. The new single-chain materials were characterized by LC-MS, H-NMR and EA. LC-MS provided Relative retention times for different C-CP homologues and side-products. Mathematical deconvolution of full-scan mass spectra revealed the presence of chloroparaffins (57-93%) and chloroolefins (COs, 7-26%) in the four single-chain C-CP-materials. Homologue distributions and chlorination degrees were deduced for CPs and COs. H-NMR revealed chemical shift ranges of mono-chlorinated (δ = 3.2-5.3 ppm) and non-chlorinated (δ = 1.0-3.2 ppm) hydrocarbon moieties. The synthesized C-single-chain standard materials and respective spectroscopic data are useful to identify and quantify LCCPs in various materials and environmental samples. CP- and CO-distributions resemble the ones of existing SCCP and MCCP reference materials and technical mixtures. Furthermore, these materials now allow specific studies on the environmental fate and the transformation of long-chain chloroparaffins and chloroolefins.
Topics: China; Environmental Monitoring; Halogenation; Hydrocarbons, Chlorinated; Mass Spectrometry; Paraffin
PubMed: 34798110
DOI: 10.1016/j.chemosphere.2021.132938 -
Journal of Microscopy Jun 2022Infrared thermography has shown after roots of grasses freeze, ice spreads into the crown and then acropetally into leaves initially through vascular bundles. Leaves...
Infrared thermography has shown after roots of grasses freeze, ice spreads into the crown and then acropetally into leaves initially through vascular bundles. Leaves freeze singly with the oldest leaves freezing first and the youngest freezing later. Visualising the vascular system in its native 3-dimensional state will help in the understanding of this freezing process. A 2 cm section of the crown that had been infiltrated with aniline blue was embedded in paraffin and sectioned with a microtome. A photograph of the surface of the tissue in the paraffin block was taken after the microtome blade removed each 20 μm section. Two hundred to 300 images were imported into Adobe After Effects and a 3D volume of the region infiltrated by aniline blue dye was constructed. The reconstruction revealed that roots fed into what is functionally a region inside the crown that could act as a reservoir from which all the leaves are able to draw water. When a single root was fed dye solution, the entire region filled with dye and the vascular bundles of every leaf took up the dye; this indicated that the vascular system of roots was not paired with individual leaves. Fluorescence microscopy suggested the edge of the reservoir might be composed of phenolic compounds. When plants were frozen, the edges of the reservoir became leaky and dye solution spread into the mesophyll outside the reservoir. The significance of this change with regard to freezing tolerance is not known at this time. Thermal cameras that allow visualisation of water freezing in plants have shown that in crops like wheat, oats and barley, ice forms first at the bottom of the plant and then moves upwards into leaves through water conducting channels. Leaves freeze one at a time with the oldest leaves freezing first and then younger ones further up the stem freeze later. To better understand why plants freeze like this, we reconstructed a 3-dimensional view of the water conducting channels. After placing the roots of a wheat plant in a blue dye and allowing it to pull the dye upwards into leaves, we took a part of the stem just above the roots and embedded it in paraffin. We used a microtome to slice a thin layer of the paraffin containing the plant and then photographed the surface after each layer was removed. After taking about 300 images, we used Adobe After Effects software to re-construct the plant with the water conducting system in three dimensions. The 3D reconstruction showed that roots fed into a roughly spherical area at the bottom of the stem that could act as a kind of tank or reservoir from which the leaves pull up water. When we put just one root in dye, the entire reservoir filled up and the water conducting channels in every leaf took up the dye. This indicates that the water channels in roots were not directly connected to specific leaves as we had thought. When plants were frozen, the dye leaked out of the reservoir and spread into cells outside. Research is continuing to understand the significance of this change during freezing. It is possible that information about this effect can be used to help breeders develop more winter-hardy crop plants.
Topics: Freezing; Ice; Paraffin; Plants; Triticum; Water
PubMed: 35319110
DOI: 10.1111/jmi.13101 -
The Cochrane Database of Systematic... Jan 2011Constipation is common in palliative care; it can generate considerable suffering due to the unpleasant physical symptoms. In the first Cochrane Review on effectiveness... (Meta-Analysis)
Meta-Analysis Review
BACKGROUND
Constipation is common in palliative care; it can generate considerable suffering due to the unpleasant physical symptoms. In the first Cochrane Review on effectiveness of laxatives for the management of constipation in palliative care patients, published in 2006, no conclusions could be drawn because of the limited number of evaluations. This article describes the first update of this review.
OBJECTIVES
To determine the effectiveness of laxatives or methylnaltrexone for the management of constipation in palliative care patients.
SEARCH STRATEGY
We searched databases including MEDLINE and CENTRAL (The Cochrane Library) in 2005 and in the update to August 2010.
SELECTION CRITERIA
Randomised controlled trials (RCTs) evaluating laxatives for constipation in palliative care patients. In the update we also included RCTs on subcutaneous methylnaltrexone; an opioid-receptor antagonist that is now licensed for the treatment of opioid-induced constipation in palliative care when response to usual laxative therapy is insufficient.
DATA COLLECTION AND ANALYSIS
Two authors assessed trial quality and extracted data. The appropriateness of combining data from the studies depended upon clinical and outcome measure homogeneity.
MAIN RESULTS
We included seven studies involving 616 participants; all under-reported methodological features. In four studies the laxatives lactulose, senna, co-danthramer, misrakasneham, and magnesium hydroxide with liquid paraffin were evaluated. In three methylnaltrexone.In studies comparing the different laxatives evidence was inconclusive. Evidence on subcutaneous methylnaltrexone was clearer; in combined analysis (287 participants) methylnaltrexone, in comparison with a placebo, significantly induced laxation at 4 hours (odds ratio 6.95; 95% confidence interval 3.83 to 12.61). In combined analyses there was no difference in the proportion experiencing side effects, although participants on methylnaltrexone suffered more flatulence and dizziness. No evidence of opioid withdrawal was found. In one study severe adverse events, commonly abdominal pain, were reported that were possibly related to methylnaltrexone. A serious adverse event considered to be related to the methylnaltrexone also occurred; this involved a participant having severe diarrhoea, subsequent dehydration and cardiovascular collapse.
AUTHORS' CONCLUSIONS
The 2010 update found evidence on laxatives for management of constipation remains limited due to insufficient RCTs. However, the conclusions of this update have changed since the original review publication in that it now includes evidence on methylnaltrexone. Here it found that subcutaneous methylnaltrexone is effective in inducing laxation in palliative care patients with opioid-induced constipation and where conventional laxatives have failed. However, the safety of this product is not fully evaluated. Large, rigorous, independent trials are needed.
Topics: Analgesics, Opioid; Anthraquinones; Cathartics; Constipation; Humans; Lactulose; Magnesium Hydroxide; Naltrexone; Palliative Care; Paraffin; Quaternary Ammonium Compounds; Randomized Controlled Trials as Topic; Senna Extract
PubMed: 21249653
DOI: 10.1002/14651858.CD003448.pub3 -
European Review For Medical and... Apr 2023Nasal polyps are non-cancerous, soft painless growth of nasal mucosa. In this study, our aim was to investigate the Ki-67 expression level in nasal polyps by...
OBJECTIVE
Nasal polyps are non-cancerous, soft painless growth of nasal mucosa. In this study, our aim was to investigate the Ki-67 expression level in nasal polyps by immunohistochemical method.
PATIENTS AND METHODS
30 patients with nasal polyps were included in this study. Nasal polyps were processed for paraffin wax embedding protocol. Samples were fixed and embedded in paraffin blocks. 5 µm sections were stained with Hematoxylin-Eosin dye and immune stained with Ki-67 antibody. Sections were analyzed under light microscope.
RESULTS
Blood parameters showed that white blood cells, hematocrit and platelet were higher than normal range. In sections of hematoxylin-eosin staining, elevated basal cells, thin basement membrane, leukocyte infiltration, collagen fibers degeneration were observed. Masson trichrome staining revealed that degenerative epithelial cells, detached basement membrane and edema were observed. Ki-67 expression was observed in mucosal epithelial cells, vascular endothelial cells and plasma cells in immune staining.
CONCLUSIONS
Epithelial degeneration in nasal polyps and leukocyte infiltration induce nasal adenoma. Ki-67 expression may be a diagnostic tool for epithelial leukocyte formation.
Topics: Humans; Nasal Polyps; Ki-67 Antigen; Endothelial Cells; Eosine Yellowish-(YS); Hematoxylin; Paraffin; Nasal Mucosa
PubMed: 37070875
DOI: 10.26355/eurrev_202304_31904 -
Environmental Science and Pollution... Jan 2023Paraffin (petroleum) oils have been used for many years for their insecticidal properties, but relatively little research had been conducted towards their introduction... (Review)
Review
Paraffin (petroleum) oils have been used for many years for their insecticidal properties, but relatively little research had been conducted towards their introduction into the agricultural praxis, due to their potential phytotoxic effects. In the recent years, however, there has been an increased interest in petroleum-based pesticides due to their compatibility with integrated pest management (IPM) programs. Various improvements in the refinement methods have enhanced the manufacture of commercial products with many advantageous features over the original oil formulas. However, literature is still lacking of a general overview about the applicability of newly introduced commercial petroleum oils in agriculture and their compatibility with modern pest management practices. Therefore, the present work aims to depict the current status of petroleum oils in arboriculture and beyond, providing an in-depth analysis of their insecticidal properties with respect to the knowledge gained over the years about the factors responsible for the pesticidal efficacy and the phytotoxic activity of petroleum-derived oil insecticides. Moreover, commercial aspects of petroleum oil formulations and their toxicological profile to non-target organisms have also been addressed through the current legislation in EU and the USA.
Topics: Paraffin; Agriculture; Oils; Pesticides; Insecticides; Petroleum
PubMed: 36396765
DOI: 10.1007/s11356-022-24059-5 -
Nephron 2023The three primary sites of acute T-cell-mediated rejection (TCMR) in transplanted kidneys are the tubular epithelial cells, interstitium, and the vascular endothelial... (Review)
Review
The three primary sites of acute T-cell-mediated rejection (TCMR) in transplanted kidneys are the tubular epithelial cells, interstitium, and the vascular endothelial cells. The pathology of acute lesions is characterized by inflammatory cell infiltration; the final diagnosis suggested by the Banff 2019 classification is guided by grading of tubulitis (the t score), interstitial inflammation (the i score), and endarteritis (the v score). Consistent major issues when using the Banff classification are the etiological classifications of interstitial fibrosis and tubular atrophy (IFTA). From 2015 to 2019, technological advances (i.e., genetic analysis in paraffin sections) increased our understanding of IFTA status in patients with smoldering acute TCMR and the roles played by inflammatory cell infiltration (the i-IFTA score) and tubulitis (the t-IFTA score) in IFTA. These two scores were introduced when establishing the diagnostic criteria for chronic active TCMR. Despite the increase in complexity and the lack of a consensus treatment for chronic active TCMR, the Banff classification may evolve as new techniques (i.e., genetic analysis in paraffin sections and deep learning of renal pathology) are introduced. The Banff conference proceeded as follows. First, lesions were defined. Next, working groups were established to better understand the lesions and to derive better classification methods. Finally, the new Banff classification was developed. This approach will continue to evolve; the Banff classification will become a very useful diagnostic standard. This paper overviews the history of TCMR diagnosis using the Banff classification, and the clinical importance, treatment, and prospects for acute and chronic active TCMR.
Topics: Humans; Kidney Transplantation; T-Lymphocytes; Endothelial Cells; Paraffin; Kidney; Kidney Diseases; Graft Rejection; Biopsy
PubMed: 36928337
DOI: 10.1159/000530158