Staining Properties Research Articles

SiglecF+Gr1hi eosinophils are a distinct subpopulation within the lungs of allergen-challenged mice.

Although eosinophils as a group are readily identified by their unique morphology and staining properties, flow cytometry provides an important means for identification of subgroups based on differential expression of distinct surface Ags. Here, we characterize an eosinophil subpopulation defined by high levels of expression of the neutrophil Ag Gr1 (CD45+CD11c-SiglecF+Gr1hi). SiglecF+Gr1hi eosinophils, distinct from the canonical SiglecF+Gr1- eosinophil population, were detected in allergen-challenged wild-type and granule protein-deficient (EPX-/- and MBP-1-/-) mice, but not in the eosinophil-deficient ΔdblGATA strain. In contrast to Gr1+ neutrophils, which express both cross-reacting Ags Ly6C and Ly6G, SiglecF+Gr1hi eosinophils from allergen-challenged lung tissue are uniquely Ly6G+ Although indistinguishable from the more-numerous SiglecF+Gr1- eosinophils under light microscopy, FACS-isolated populations revealed prominent differences in cytokine contents. The lymphocyte-targeting cytokines CXCL13 and IL-27 were identified only in the SiglecF+Gr1hi eosinophil population (at 3.9 and 4.8 pg/106 cells, respectively), as was the prominent proinflammatory mediator IL-13 (72 pg/106 cells). Interestingly, bone marrow-derived (SiglecF+), cultured eosinophils include a more substantial Gr1+ subpopulation (∼50%); Gr1+ bmEos includes primarily a single Ly6C+ and a smaller, double-positive (Ly6C+Ly6G+) population. Taken together, our findings characterize a distinct SiglecF+Gr1hi eosinophil subset in lungs of allergen-challenged, wild-type and granule protein-deficient mice. SiglecF+Gr1hi eosinophils from wild-type mice maintain a distinct subset of cytokines, including those active on B and T lymphocytes. These cytokines may facilitate eosinophil-mediated immunomodulatory responses in the allergen-challenged lung as well as in other distinct microenvironments.

Isolation, molecular identification and antibiotic susceptibility profile of Aeromonas hydrophila from cultured indigenous Koi (Anabus testudineus) of Bangladesh

Fish play a crucial role in the Bangladeshi diet, providing more than 60% of animal source food, representing a crucial source of micro-nutrients and possessing an extremely strong cultural attachment. In this study isolation and identification of Aeromionas hydrophila was done by studying cultural properties, Gram’s staining and biochemical properties of isolates of diseased indigenous Koi fish (Anabas testudineus) of different upazillas of Mymensingh district. Antibiogram profile of the isolated bacteria was studied by using wide range of commercially available antibiotics. Quantitative study of bacteria isolated from diseased indigenous Koi fish showed variation of number in different organ. Total bacterial load was found to be 1.90 × 105, 1.19 × 105, 3.21 × 105, 2.18 × 106 and 3.14 × 105 cfu/g in lesions; 2.52 × 107, 2.34 × 108, 5.41 × 108, 2.54 × 109 and 5.21 × 109 cfu/g in liver; 2.54 × 108, 2.41 × 108, 1.90 × 107, 3.65 × 107 and 3.45 × 108 cfu/g in spleen; 3.51 × 107, 5.28 × 107, 3.14 × 106, 1.85 × 107 and 4.52× 107 cfu/g in kidney in diseased Koi of Mymensingh sadar, Muktagacha, Tarakanda, Gouripur and Fulpur upazillas, respectively under Mymensingh districts. Aeromonas hydrophila was initially identified by their specific morphological, physiological and biochemical characteristics. Then molecular detection of A. hydrophila was done by PCR. PCR products of desired 760 bp were obtained for A. hydrophila. The results of the antibiotic sensitivity test is exhibited that most of the bacterial samples were sensitive against ciprofloxacin (92%) and levofloxacin (84%), intermediate resistant against gentamicin (40%) and resistant against novobiocin (84%), ampicillin (100%) and penicillin (92%).Asian J. Med. Biol. Res. June 2016, 2(2): 332-340

HBME-1 is expressed by erythroid precursors in early maturation stage and can be a valuable tool for evaluation of dyserythropoiesis in bone marrow core biopsy specimens.

The reaction of Hector Battifora mesothelial epitope-1 (HBME-1) antibody with scattered pronormoblasts in normal bone marrow core biopsy specimens has been reported. This study evaluated the immunohistochemical profile of HBME-1 in a panel of 52 normal, dyserythropoietic and neoplastic marrow samples. We compared the staining property of HBME-1 with that of the commonly used erythroid marker, glycophorin A (CD235a) and in each case, we semi-quantitatively evaluated the HBME-1/CD235a-positive cells ratio. In normal samples, HBME-1 labelled scattered immature erythroid precursors. In dyserythropoietic specimens, HBME-1 stained nucleated erythroid precursors in varying degrees, from pronormoblast through normoblast stages, with the highest intensity in immature forms. Overall, the cellular background of non-erythroid progenitors, erythrocytes and neoplastic cells did not react with HBME-1, except in leukaemia cases with myelodysplasia-related changes. Our study shows that HBME-1 is a useful marker to identify immature erythroid precursors and that an HBME-1/CD235a-positive cells ratio ≥10% is associated with dyserythropoiesis.

A Red to Near-IR Fluorogen: Aggregation-Induced Emission, Large Stokes Shift, High Solid Efficiency and Application in Cell-Imaging.

A tetraphenylethene (TPE) derivative modified with the strong electron acceptor 2-dicyano-methylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran (TCF) was obtained in high yield by a simple two-step reaction. The resultant TPE-TCF showed evident aggregation-induced emission (AIE) features and pronounced solvatochromic behavior. Changing the solvent from apolar cyclohexane to highly polar acetonitrile, the emission peak shifted from 560 to 680 nm (120 nm redshift). In an acetonitrile solution and in the solid powder, the Stokes shifts are as large as 230 and 190 nm, respectively. The solid film emits red to near-IR (red-NIR) fluorescence with an emission peak at 670 nm and a quantum efficiency of 24.8 %. Taking the advantages of red-NIR emission and high efficiency, nanoparticles (NPs) of TPE-TCF were fabricated by using tat-modified 1,2-distearoylsn-glycero-3-phosphor-ethanol-amine-N-[methoxy-(polyethyl-eneglycol)-2000] as the encapsulation matrix. The obtained NPs showed perfect membrane penetrability and high fluorescent imaging quality of cell cytoplasm. Upon co-incubation with 4,6-diamidino-2-phenylindole (DAPI) in the presence of tritons, the capsulated TPE-TCF nanoparticles could enter into the nucleus and displayed similar staining properties to those of DAPI.

Peculiarities of human pancreatolienal hemolymph node structure and cellular composition.

Background. Hemolymph nodes (HLNs) are well described in animals, while in humans they are still poorly characterized; little is known about the role of HLNs in red blood cells destruction, erythropoiesis, iron metabolism etc. Objective. To determine the peculiarities of HLN structure and cellular composition. Methods. Pancreatolienal HLNs were taken during autopsy from patients who died due to cardiovascular or respiratory pathology. Standard histological and histochemical methods were used to detect iron, bile components, lipofuscin. Results. Inside afferent vessels of HLNs equal numbers of red and white blood cells were found. The same ratio was in subcapsular and cortical sinuses, while in medullary sinus erytrhrocytes were absent. Some RBCs enter lymphoid follicles through opened blood capillaries. Inside HLNs parenchyma large lipofuscin-laden cells are frequently observed, by their staining properties they resemble sea-blue histiocytes found under normal circumstances in red bone marrow. Conclusions. HLNs receive mixture of blood and lymph; during circulation through the system of sinuses all erythrocytes are destroyed via phagocytosis by local macrophages. During hemoglobin degradation these macrophages release iron outside, but undigested membranes of RBCs undergo lipid peroxidation giving rise to lipofuscin stored inside the cell.

MUSE: A New, Fast, Simple Microscopy Method for Slide‐Free Histology and Surface Topography

IntroductionConventional brightfield or fluorescence microscopy works best with thin, stained specimens mounted on glass slides, but to prepare these requires hours of processing and the help of highly skilled technical personnel. We describe a new, inexpensive form of light microscopy that can generate high‐quality histology and histopathology images directly from cut surfaces of fresh (or fixed) tissue samples of any thickness, with less than 1 minute of preparation.BackgroundWhile alternative microscopy‐scale methods, such as confocal or two‐photon, or optical coherence tomography approaches, can generate reasonable images from thick tissues, the instrumentation is typically complex, expensive and not widely available. We are developing an alternative technique, MUSE (Microscopy with UV Surface Excitation), that relies on simple properties of light, tissue, and fluorescent stains, and can be implemented at relatively low cost. Thick pieces of tissue obtained by biopsy, surgery or necropsy can be imaged directly. The method is non‐destructive, and eliminates requirement for conventional histology processing, formalin fixation, paraffin‐embedding, or thin‐sectioning.MethodUltraviolet light at about 280 nm from an LED arranged to provide oblique illumination, is used to excite just the surface layer of tissue that has been briefly (~10 seconds) stained with common and inexpensive fluorescent dyes. Unlike light of longer wavelength, 280‐nm light only penetrates to a depth of 10 microns or less, and thus excites fluorescent signals, conveniently in the visible range, only from the cut specimen surface. The images, which can be diffraction‐limited, are captured using conventional microscope optics and a standard color cameraResultsMUSE samples from fresh (or fixed) tissues stained with general tissue stains such as rhodamine and Hoechst generate high‐resolution fluorescence images that can be converted in real time from fluorescence to H&E‐like brightfield appearance, for ease of interpretation. However, fluorescence display mode preserves surface shading and depth cues that allow for appreciation of surface profiles important in understanding 3D organization of complex specimens. We have obtained high‐quality MUSE images from normal and diseased tissues from multiple organisms, as well as from plants, and manufactured materials. Moreover, surface profile information provides views beyond what can be seen with in standard thin‐sectioned material, generating results that can resemble those obtainable with scanning electron microscopy (SEM).ConclusionWe have developed a new form of optical microscopy that generates diagnostic‐quality histological images, with enhanced content, from fresh or fixed, but unsectioned tissue, rapidly, with high resolution, simply and inexpensively. We anticipate that there could be widespread adoption in preclinical and clinical research facilities, as well as hospital‐based and stand‐alone medical settings.

Synthesis and Preliminary Biological Evaluation of Fluorescent Glycofused Tricyclic Derivatives of Amyloid β‐Peptide Ligands

AbstractFluorescent glycofused tricyclic compounds were synthesized through the domino conjugate oxa‐Michael addition/aldol condensation of 2‐hydroxybenzaldehydes with protected 3‐oxoglucal by exploiting activation with an organocatalyst. Stereoselectivity was obtained by using (R)‐(+)‐α,α‐diphenyl‐2‐pyrrolidinemethanol trimethylsilyl ether as the catalyst. All synthesized compounds presented good optical properties. All compounds were able to bind to synthetic amyloid β 1–42 peptide aggregates and to label amyloid plaques from brain sections of transgenic mice affected with Alzheimer's disease with staining properties comparable to thioflavin T.

Effects of the Aqueous Extract of Aloe vera on the Morphological and Physiological Properties of E. coli

Effects of the Aqueous Extract of Aloe vera on the Morphological and Physiological Properties of E. coli ________________________________________ Mahdi Kargaran 1 ; Ali Reza Moradabadi 1 ; Mohammad Arjomandzadegan 1, * ; Hossein Hosseini 1 ; Ghasem Habibi 1 ; Maryam Tayeboon 1 ; Hadi Karami 1 ; and Ahmad Akbari 1, * 1 Tuberculosis and Pediatric Infectious Research Center, Department of Microbiology, Arak University of Medical Sciences, Arak, IR Iran *Corresponding Authors: Mohammad Arjomandzadegan, Tuberculosis and Pediatric Infectious Research Center, Department of Microbiology, Arak University of Medical Sciences, Arak, IR Iran, E-mail: arjomandzadegan@arakmu.ac.ir; Ahmad Akbari, Tuberculosis and Pediatric Infectious Research Center, Department of Microbiology, Arak University of Medical Sciences, Arak, IR Iran, E-mail: dr.akbari@arakmu.ac.ir. Abstract Background: Resistance to antibiotics is a growing worldwide problem. The inappropriate use of antibiotics has rendered some strains of bacteria resistant to antimicrobial drugs, making the treatment of infections much more difficult. However, many herbs and their derivatives are effective against drug resistant microbes. Objectives: The aim of this research was to evaluate the effects of an Aloe vera extract on the morphological and physiological properties of Escherichia coli. Patients and Methods: A clinical isolate of E. coli was obtained from a patient harboring a urinary tract infection (UTI), and was identified using biochemical and microbiological methods. Aqueous extracts of Aloe vera were prepared, and the minimum inhibitory concentration (MIC) of the extract was determined for E. coli via the microdilution method. The bacterium, at a concentration of 1.5 × 108 cells/mL, was exposed to the MIC of the extract. Then, the morphology of the cells was studied using optical microscopy, and the physiological properties were studied using biochemical testing and differential culture media. Results: The MIC of the Aloe vera extract was 2.23 mg/mL, and was able to prevent the growth of E. coli. The morphological examination of the bacteria exposed to the extract at the MIC revealed that the cells were shrunken, the concentration was reduced, the number of coccobacilli was increased, and the staining property of this bacteria changed (P < 0.0001). In addition, the extract caused a 76% reduction in the bacterial cell number, in comparison to the control (P < 0.0001), and a change in the physiological properties and growth of the bacteria. Conclusions: This study showed that Aloe vera has antibacterial effects against E. coli, and can result in structural and physiological changes in this bacterium. Keywords: Morphological Changes; Physiological Changes; Aloe vera; E. coli

The Impact of the Fourth Edition of the WHO Classification of Lung Tumours on Histological Classification of Resected Pulmonary NSCCs

IntroductionHistopathological classification of lung cancer is of central importance in the diagnostic routine, and it guides therapy in most patients. The fourth edition of the World Health Organization (WHO) Classification of Lung Tumours was recently published and includes changes to the diagnostic procedure for non–small cell carcinomas (NSCCs), with more emphasis on immunohistochemical (IHC) staining. MethodsA total of 656 unselective cases of resected pulmonary NSCC were diagnosed according to the 2004 WHO classification. After IHC staining with cytokeratin 5, p40, p63, thyroid transcription factor 1 (clones 8G7G3/1 and SPT24), and napsin A, the diagnoses were revised in accordance with the new fourth edition of the WHO classification. ResultsReclassification led to a new histological annotation in 36 of the 656 cases (5%). Most notable was the decrease in cases previously classified as large cell carcinomas (56 versus 12 cases). This was partially due to the exclusion of 21 neuroendocrine tumors from this group, with 20 cases ascribed to the adenocarcinoma group on the basis of IHC markers. Only seven cases of adenocarcinoma or squamous cell carcinoma were reclassified after the addition of IHC staining. There was a substantial overlap in staining properties between different markers of squamous and adenocarcinomatous differentiation, respectively, but in 17 to 31 cases (3%–5%), the diagnosis depended on the choice of markers. ConclusionsThe fourth edition of the WHO Classification of Lung Tumours leads to changes in histological type in 5% of resected NSCCs. The incorporation of IHC staining into NSCC diagnostics demands awareness that the choice of ancillary stains has an effect on diagnosis.

Molecular Engineering of Thiazole Orange Dye: Change of Fluorescent Signaling from Universal to Specific upon Binding with Nucleic Acids in Bioassay.

The universal fluorescent staining property of thiazole orange (TO) dye was adapted in order to be specific for G-quadruplex DNA structures, through the introduction of a styrene-like substituent at the ortho-position of the TO scaffold. This extraordinary outcome was determined from experimental studies and further explored through molecular docking studies. The molecular docking studies help understand how such a small substituent leads to remarkable fluorescent signal discrimination between G-quadruplex DNA and other types of nucleic acids. The results reveal that the modified dyes bind to the G-quadruplex or duplex DNA in a similar fashion as TO, but exhibit either enhanced or quenched fluorescent signal, which is determined by the spatial length and orientation of the substituent and has never been known. The new fluorescent dye modified with a p-(dimethylamino)styryl substituent offers 10-fold more selectivity toward telomeric G-quadruplexes than double-stranded DNA substrates. In addition, native PAGE experiments, FRET, CD analysis, and live cell imaging were also studied and demonstrated the potential applications of this class of thiazole-orange-based fluorescent probes in bioassays and cell imaging.

The effect of dietary pigmentation on the esthetic appearance of clear orthodontic elastomeric modules.

Objective:To compare the stain resistance of three types of clear elastomeric modules exposed to several common dietary substances through the assessment of the perception of a group of dentists to discoloration using visual analog scale (VAS).Materials and Methods:Elastomeric modules from Unitek (AU), Ormco (OR), and dentaurum (DE) were immersed in the following food substances: Coffee, black tea, chocolate, energy drink, ketchup, and Coca-Cola for 72 h. VAS was used to reflect the module staining severity.Results:Significant difference was found among the three types of modules examined in this study. OR modules showed the least mean staining ratings by the examiners. There was no statistical difference in the staining properties between AU and DE modules. Coffee and tea showed higher staining potential as compared to all staining media. Furthermore, there was no difference in the staining characteristics of coffee and black tea.Conclusions:Coffee and tea are strong staining media that should be avoided by patients who opted to have esthetic appliances for their orthodontic treatment. Elastomeric modules manufactured by AU showed higher staining optical properties as compared to the other two companies, which could be related to the manufacturing processing of these modules.

Ectropion and Conjunctival Mass in a Patient with Primary Bilateral Conjunctival Amyloidosis.

Background. Amyloidosis is a group of disorders characterized by deposition of an extracellular protein, known as amyloid, in an abnormal fibrillar form with highly characteristic histopathologic staining properties. The clinical presentation can vary from a focal, localized lesion where amyloidosis has minor clinical consequences to extensive systemic disease that can involve any organ system of the body. Ocular amyloidosis can occur as a localized lesion or as a part of a systemic disorder. Conjunctival amyloidosis is an uncommon condition that is rarely associated with systemic disease. It may be a manifestation of an immunologic disorder. Case Report. We report the case of a patient with bilateral conjunctival amyloidosis who was referred to us with the suspicion of a malignant conjunctival lesion. Examination of both eyes showed a yellow-pink mass with prominent intrinsic vessels, subconjunctival hemorrhage, and ectropion of the left lower eyelid. Diagnosis of primary localized conjunctival amyloidosis was made based on histopathologic evaluation of incisional biopsy and negative systemic work-up. Conclusion. Ocular amyloidosis is a rare disease that is slowly progressive and has a wide variety of clinical presentations. Consequently, the clinical diagnosis is often overlooked or delayed. Definitive diagnosis is achieved through histopathologic evaluation of biopsy specimen.

DNA-histone complexes as ligands amplify cell penetration and nuclear targeting of anti-DNA antibodies via energy-independent mechanisms.

We have generated three monoclonal cell-penetrating antibodies (CPAbs) from a non-immunized lupus-prone (NZB × NZW)F1 mouse that exhibited high anti-DNA serum titres. These CPAbs are polyreactive because they bind to DNA and other cellular components, and localize mainly in the nucleus of HeLa cells, albeit with a distinct nuclear labelling profile. Herein, we have examined whether DNA-histone complexes (DHC) binding to CPAbs, before cell entry, could modify the cell penetration of CPAbs or their nuclear staining properties. By applying confocal microscopy and image analysis, we found that extracellular binding of purified CPAbs to DHC significantly enhanced their subsequent cell-entry, both in terms of percentages of positively labelled cells and fluorescence intensity (internalized CPAb amount), whereas there was a variable effect on their nuclear staining profile. Internalization of CPAbs, either alone or bound to DHC, remained unaltered after the addition of endocytosis-specific inhibitors at 37° or assay performance at 4°, suggesting the involvement of energy-independent mechanisms in the internalization process. These findings assign to CPAbs a more complex pathogenetic role in systemic lupus erythematosus where both CPAbs and nuclear components are abundant.

Do we need to demonstrate amyloid in tissue for hereditary ATTR amyloidosis ?

Amyloidosis is by definition deposition of aggregates of proteins in a characteristic β-pleated sheet fibrillar conformation. Amyloid is recognized by its histological appearance particularly after some specific stainings among which Congo red most commonly is used. A green birefringence is the hallmark of all kinds of amyloid. There is a number of emerging diagnostic techniques based on labelling with antibodies, other proteins and some other ligands that are at least to some degree specific for amyloid and may be used for in vivo detection of amyloid. With some exceptions, such techniques are still not sensitive enough to detect small deposits. Biopsy stained with Congo red and examined in a polarization microscope is necessary presently. Hereditary ATTR amyloidosis constitutes a heterogeneous disease group with varying organ consequences, not only from peripheral nerves and heart. Amyloid staining properties vary and Congo red affinity and birefringence may be very weak, particularly in amyloid consisting of TTR fragments which is common (1). Therefore is amyloid sometimes difficult to recognize also in biopsies. Given the varying clinical manifestation of ATTR amyloidosis it is my opinion that a histological proof should be obtained if possible. There may be exceptions sometimes depending on local conditions. In endemic areas with one specific TTR mutation, high penetrance and a relative monomorphous presentation of ATTR amyloidosis, diagnosis may sometimes be safe without a histological proof. This may be the case in some areas of Portugal. In most parts of the world, however, cases of systemic amyloidosis are usually scattered and a histological proof should be recommended. The site of biopsy may be discussed. We perform subcutaneous adipose tissue biopsies (surgical or by punch technique) with good results. Fine needle biopsy is not recommended since with this technique very little connective tissue is obtained, which is the site of ATTR amyloid containing TTR fragments (2). In other centers, labial salivary gland biopsies are preferred. Gastrointestinal tract is another option. A substantial advantage with biopsy techniques is that the biochemical type can be determined directly. A question to discuss is whether indirect techniques such as DPD scintigraphy, SAP scintigraphy or PET with various ligands are safe enough for a definite amyloid diagnosis. Also alternatives to Congo red are under development but not yet validated enough for clinical use.

Combined Spectroscopic And Molecular Docking Study Of Binding Interaction Of Pyrano [3, 2-f] Quinoline Derivatives With Bovine Serum Albumins And Its Application In Mammalian Cell Imaging

The interaction between pyrano [3, 2-f] quinoline derivatives (TPQ) and bovine serum albumin (BSA) was studied using spectroscopic techniques. The TPQ quench the fluorescence of BSA through dynamic quenching. According to Van’t Hoff equation, the thermodynamic parameters were calculated and which indicated hydrogen bonds and van der waals forces played a prime role in stabilizing the BSA–TPQ complexes. Also, the average binding distance (r) and the critical energy transfer distance (Ro) between TPQ and BSA were also evaluated according to Förster’s non-radiative energy transfer (FRET) theory. What is more, UV-visible and circular dichroism results showed that the addition of TPQ changed the secondary structure of BSA and led to a reduction in content α-helix (%) content. It was also observed that TPQ shows cell staining property to the cultured HeLa cell line. Theoretical docking study of interaction between BSA and TPQ also supported the experimental results. All the results suggested that BSA experienced substantial conformational changes induced by TPQ; this may be useful to study synthetic organic molecules for their application as pharmaceuticals.

Accuracy of a New Platelet Count System (PLT-F) Depends on the Staining Property of Its Reagents.

BackgroundPlatelet count is essential for the diagnosis and management of hemostasis abnormalities. Although existing platelet count methods installed in common hematology analyzers can correctly count platelets in normal blood samples, they tend to miscount platelets in some abnormal samples. The newly developed PLT-F channel in the XN-Series hematology analyzer (Sysmex) has been reported to be a reliable platelet count system, even in abnormal samples. However, how the PLT-F platelet counting system achieves such accuracy has not been described in scientific articles.MethodsIsolated platelets, erythrocytes, and fragmented erythrocytes were examined using an automated hematology analyzer. The samples were labeled by combining PLT-F reagents and anti-CD62p, CD63, Grp75, Calreticulin, CD41, or CD61 antibody, and analyzed using confocal laser scanning microscopy or flow cytometry.ResultsThe PLT-F system correctly discriminated platelets in erythrocytes. Its reagents strongly stained some intraplatelet organelles labeled with anti-Grp75, but only faintly stained the plasma membrane of both platelets and erythrocytes. Microscopic observation and flow cytometric examination revealed that all of these strongly stained cells were also labeled with platelet-specific anti-CD41 and anti-CD61 antibodies.ConclusionsThis study revealed that the staining property of the PLT-F reagents, by which platelets and fragmented erythrocytes are clearly distinguished, contributes to the platelet-counting accuracy of the PLT-F system.

Tight junction defects are seen in the buccal mucosa of patients receiving standard dose chemotherapy for cancer.

Oral mucositis is one of the most common and debilitating side effects of chemotherapy treatment. Patients are often unable to eat and drink, which can lead to poor clinical outcomes and extensive resource utilisation. The primary aim of this study was to determine the molecular integrity of oral epithelial tight junctions in patients undergoing chemotherapy. The secondary aim was to correlate these changes with proinflammatory cytokines and matrix metalloproteinase profiles. Patients (n = 23) were recruited from the Royal Adelaide Hospital between 2000 and 2003. Reach patient underwent two oral buccal mucosa biopsies (4 mm): one prior to chemotherapy treatment and a second one after chemotherapy treatment. Oral buccal mucosa biopsies were also taken from seven healthy volunteers with no history of cancer, chemo- or radiotherapy treatment or inflammatory disorders. Routine haematoxylin and eosin staining was performed to determine epithelial thickness. Immunohistochemical staining was performed for claudin-1, zonular occludens-1, occludin, interleukin-1β, tumour necrosis factor, interleukin-6, matrix metalloproteinase-2 and metalloproteinase-9. Patients receiving standard dose chemotherapy had significant epithelial atrophy. Elevations in all cytokines and matrix metalloproteinases were seen, with significant lamina propria staining for interleukin-6 and tumour necrosis factor. Matrix metalloproteinase-2 appeared most upregulated within the oral epithelium. These changes coincided with altered tight junction staining properties. Changes in the staining intensity and localisation were both noted, with clear cytoplasmic staining for zonular occludens-1 and claudin-1 in patients treated with chemotherapy. Chemotherapy causes defects in oral tight junctions, coupled with altered cytokine and matrix metalloproteinase profiles. Tight junction disruption in the epithelium may contribute to ulcer development or lead to poor tissue integrity, and the timing of these events may be a target for preventative treatment.

Amyloid P Immunohistochemistry in Myocardial Amyloidosis: Comparison With Congo Red

Amyloid refers to a family of abnormal proteins that share a common fibrillar β-pleated sheet structure that gives it unique optical and staining properties. In addition to the fibrillar component, amyloid deposits contain a universal amyloid P component. Detection of amyloid in myocardial tissue with Congo red alone is difficult and tedious. We evaluated …

Improvement of the standard method for assessing colour fastness to wet rubbing

Colour fastness to rubbing is evaluated by the degree of staining of a standard adjacent cotton cloth. However, some kinds of dye usually stain other textiles to a greater extent than cotton under wet conditions. In this work, rubbing fastness tests were carried out with various kinds of adjacent cloth in order to examine the factors affecting the wet rubbing fastness, and a more practical evaluation method using adjacent rubbing cloth was proposed. Dye transfer from the dyed fabric to the adjacent cloth depended on the affinity of the dyes for the fibre material of the dyed fabric and the adjacent cloth under wet conditions, in addition to the friction coefficient between the fabrics. A standard adjacent cloth should have a high staining property. The nylon staple cloth is preferred for fabrics dyed with vat, disperse, and acid dyes because it receives the most dye from rubbing with them.

A Historical Perspective on the Identification of Cell Types in Pancreatic Islets of Langerhans by Staining and Histochemical Techniques.

Before the middle of the previous century, cell types of the pancreatic islets of Langerhans were identified primarily on the basis of their color reactions with histological dyes. At that time, the chemical basis for the staining properties of islet cells in relation to the identity, chemistry and structure of their hormones was not fully understood. Nevertheless, the definitive islet cell types that secrete glucagon, insulin, and somatostatin (A, B, and D cells, respectively) could reliably be differentiated from each other with staining protocols that involved variations of one or more tinctorial techniques, such as the Mallory-Heidenhain azan trichrome, chromium hematoxylin and phloxine, aldehyde fuchsin, and silver impregnation methods, which were popularly used until supplanted by immunohistochemical techniques. Before antibody-based staining methods, the most bona fide histochemical techniques for the identification of islet B cells were based on the detection of sulfhydryl and disulfide groups of insulin. The application of the classical islet tinctorial staining methods for pathophysiological studies and physiological experiments was fundamental to our understanding of islet architecture and the physiological roles of A and B cells in glucose regulation and diabetes.