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1.
Medicina (Kaunas) ; 60(4)2024 Mar 30.
Article in English | MEDLINE | ID: mdl-38674213

ABSTRACT

Background and Objectives: There are many surgical techniques for oroantral communication treatment, one of which is the buccal fat pad. Of particular interest is the high reparative potential of the buccal fat pad, which may be contributed to by the presence of mesenchymal stem cells. The purpose of this work is to evaluate the reparative potential of BFP cells using morphological and immunohistochemical examination. Materials and Methods: 30 BFP samples were provided by the Clinic of Maxillofacial and Plastic Surgery of the Russian University of Medicine (Moscow, Russia) from 28 patients. Morphological examination of 30 BFP samples was performed at the Institute of Clinical Morphology and Digital Pathology of Sechenov University. Hematoxylin-eosin, Masson trichrome staining and immunohistochemical examination were performed to detect MSCs using primary antibodies CD133, CD44 and CD10. Results: During staining with hematoxylin-eosin and Masson's trichrome, we detected adipocytes of white adipose tissue united into lobules separated by connective tissue layers, a large number of vessels of different calibers, as well as the general capsule of BFP. The thin connective tissue layers contained neurovascular bundles. Statistical processing of the results of the IHC examination of the samples using the Mann-Whitney criterion revealed that the total number of samples in which the expression of CD44, CD10 and CD133 antigens was confirmed was statistically significantly higher than the number of samples where the expression was not detected (p < 0.05). Conclusions: During the morphological study of the BFP samples, we revealed statistically significant signs of MSCs presence (p < 0.05), including in the brown fat tissue, which proves the high reparative potential of this type of tissue and can make the BFP a choice option among other autogenous donor materials when eliminating OAC and other surgical interventions in the maxillofacial region.


Subject(s)
Adipose Tissue , Azo Compounds , Cheek , Immunohistochemistry , Humans , Immunohistochemistry/methods , Female , Male , AC133 Antigen/analysis , Hyaluronan Receptors/analysis , Neprilysin/analysis , Mesenchymal Stem Cells , Adult , Eosine Yellowish-(YS) , Hematoxylin , Methyl Green
2.
Trop Doct ; 54(2): 139-146, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38311979

ABSTRACT

Permanent stains such as trichrome have better sensitivity but are time-consuming and the fixative includes toxic mercuric chloride. Thus, a newer modification was tested and found to be a superior, faster and safer staining technique for intestinal parasitic detection. Our study lasted 9 months and a single stool sample was collected from each enrolled patient. We evaluated classical trichrome (T1 - using Schaudinn fixative) with newer modifications, which involved different fixatives with mordant combinations (T2 - acetic acid + hydrated aluminium sulphate, T3 - citric acid + copper sulphate hydrate). Conventional PCR targeting Entamoeba histolytica, Giardia lamblia and Cryptosporidium spp. was taken as the reference. Out of 175 stool samples, 25.1% protozoa were identified by wet mount, 24% by each T1 and T2, 25.7% by T3. Statistically, T3 and T2 had higher sensitivity as compared to T1 and wet mount when PCR was used as reference.


Subject(s)
Azo Compounds , Cryptosporidiosis , Cryptosporidium , Entamoeba histolytica , Eosine Yellowish-(YS) , Intestinal Diseases, Parasitic , Methyl Green , Parasites , Animals , Humans , Fixatives , Feces/parasitology , Intestinal Diseases, Parasitic/parasitology , Entamoeba histolytica/genetics , Coloring Agents
3.
Expert Rev Respir Med ; 17(11): 1079-1089, 2023.
Article in English | MEDLINE | ID: mdl-38058175

ABSTRACT

BACKGROUND: Diagnosis of pleural tuberculosis (TB) is tedious owing to its close resemblance with malignant pleural effusion and sparse bacterial load in clinical specimens. There is an immediate need to design a rapid and dependable diagnostic test to prevent unnecessary morbidity/mortality. RESEARCH DESIGN AND METHODS: A multi-targeted loop-mediated isothermal amplification (MT-LAMP) was deliberated using mpt64 and IS6110 to diagnose pleural TB within pleural fluids/biopsies. MT-LAMP products were analyzed by gel-based and visual detection methods, viz. SYBR Green I, SYBR Green I+deoxyuridine triphosphate uracil-N-glycosylase (dUTP-UNG), and dry methyl green reactions. RESULTS: In a pilot study, while assessing pleural TB/non-TB control subjects (n = 40), both SYBR Green I+dUTP-UNG/gel-based MT-LAMP assays exhibited better sensitivity/specificity than SYBR Green I and dry methyl green MT-LAMP. Since it is facile to work with SYBR Green I+dUTP-UNG than gel-based MT-LAMP, we validated the performance of SYBR Green I+dUTP-UNG in a higher number of specimens (n = 97), which revealed somewhat higher sensitivity (85.2 vs. 81.5%) and specificity (97.7 vs. 90.7%) than SYBR Green I MT-LAMP. Furthermore, the sensitivity attained by SYBR Green I+dUTP-UNG MT-LAMP was significantly higher (p < 0.001) than GeneXpert. CONCLUSIONS: Our SYBR Green I+dUTP-UNG MT-LAMP is a simple and reliable method to diagnose pleural TB, which may translate into a point-of-care test.


Subject(s)
Mycobacterium tuberculosis , Tuberculosis, Pleural , Humans , Tuberculosis, Pleural/diagnosis , Methyl Green , Pilot Projects , Sensitivity and Specificity , Mycobacterium tuberculosis/genetics
4.
Biosystems ; 232: 105011, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37657748

ABSTRACT

Qualia have traditionally been considered difficult to measure objectively, but with the recent spread of fMRI (functional Magnetic Resonance Imaging) and other techniques, various experimental efforts have been made. In this paper, focusing on the qualia for color, we created 6 colors with different RGB values for reference colors of RED, light GREEN, BLUE, YELLOW, and PURPLE, and showed them to 306 subjects. For example, for RED and 5 generated colors, we asked them, "Choose a color that you feel is RED," and asked them to choose. A probability density function was defined for each of the five generated colors and the reference color, which is the primary color of RED, light GREEN, BLUE, YELLOW, and PURPLE, and the Kullback-Leibler divergence between the probability density function of the reference color and the generated color was calculated, the relationship between the number of samples of the selected color and the Kullback-Leibler divergence was obtained, and the difference in color sensation-qualia was calculated accordingly. As a result, it was confirmed that the larger the distance of the Kullback-Leibler divergence, the smaller the number of samples, but that the distribution shape in which the number of samples decreased for each color differed greatly. This suggests that if we see a color such as RED to PURPLE, we are randomly choosing a color that "feels."


Subject(s)
Emotions , Methyl Green , Humans , Likelihood Functions , Mental Processes
5.
Indian J Pathol Microbiol ; 66(2): 400-402, 2023.
Article in English | MEDLINE | ID: mdl-37077096

ABSTRACT

Chronic intestinal pseudo-obstruction (CIPO) is caused by a plethora of primary and secondary causes, dominantly involving the neuromuscular tissue, interstitial cells of Cajal, or the connective tissue framework. The lack of the connective tissue framework, known as desmosis, is evaluated by Masson's trichrome (MT) or picrosirius red stains, both of which are recommended in the London classification. We evaluated the orcein stain in detecting desmosis in comparison to the MT stain. We performed both orcein and MT stains in six previously published cases of complete or partial desmosis along with six age-matched controls. Our results showed comparable results of the orcein stain as compared to the MT stain. Additional advantages of lower cost and a clearer background in orcein stain were noteworthy, whereas MT stain can be used for the detection of additional pathology. We believe that orcein stain can be used as a cheap alternative in resource-limited settings.


Subject(s)
Intestinal Pseudo-Obstruction , Methyl Green , Humans , Staining and Labeling , Intestinal Pseudo-Obstruction/diagnosis , Chronic Disease
6.
PLoS One ; 18(4): e0284165, 2023.
Article in English | MEDLINE | ID: mdl-37036886

ABSTRACT

INTRODUCTION: Health workers' failure to adhere to guidelines for screening, diagnosis and management of HIV-associated cryptococcal meningitis (CM) remains a significant public health concern. We aimed to assess adherence to the standards of care and management of HIV patients at risk of CM per the MoH guidelines and assess stock management of CM supplies in the period of January to June 2021 at selected public health facilities (HFs) in Uganda. METHODS: The study employed an observational cross-sectional design to assess the level of adherence of health workers to standards of clinical care and management of HIV positive patients at risk of CM as per the clinical guidelines for Uganda, and stock management of CM supplies in the period of January to June 2021in selected public health facilities. The study team used a survey guide designed by MoH to assess and score the screening, diagnosis and management practices of Health Facilities towards CM. Scoring was categorized as red (< 80%), light green (80%-95%), and dark green (˃95%) in the order from worst to best adherence. The data was transcribed into a spread sheet and analysed using STATA-v15. RESULTS: The study team visited a total of 15 public health facilities including 5 general hospitals, 9 regional referral hospitals (RRHs) and 1 National Referral hospital (NRH). The mean score for adherence to screening and management of CM for all the combined facilities was 15 (64.7%) classified as red. 10 (66.7%) HFs had not performed a baseline CD4 test for eligible patients within 2 weeks of ART initiation. With regards to treatment, 9 (60%) of the HFs were scored as light green on knowledge of the procedure for reconstituting intravenous Liposomal Amphotericin B. None of the HFs visited had potassium chloride tablets in stock. CONCLUSION: Major MoH guidelines are generally not being adhered to by health workers while managing cryptococcal meningitis. It is vital that government and implementing partners regularly support HFs with training, mentorship, and support supervision on CM management to improve adherence to CM screening and treatment guidelines.


Subject(s)
HIV Infections , Meningitis, Cryptococcal , Humans , Meningitis, Cryptococcal/diagnosis , Meningitis, Cryptococcal/drug therapy , HIV Infections/complications , HIV Infections/diagnosis , Uganda , Cross-Sectional Studies , Methyl Green
7.
Int J Low Extrem Wounds ; 22(3): 454-458, 2023 Sep.
Article in English | MEDLINE | ID: mdl-33998845

ABSTRACT

MolecuLight i:X is a noninvasive, portable device that captures images, measures wound areas, and allows the evaluation of the bacterial environment in real time. The aim of the study was to correlate the different fluorescence (light green, red, cyan) and dark red-purple-black color areas with average pH values in these areas and with average wound bed score (WBS). During a 4-week period, we enrolled 43 adult patients (23 females and 20 males) with clinically infected and uninfected chronic ulcers. In our study, the mean age was 68 years old. The etiologies were 21 venous ulcers, 3 arterial ulcers, 4 vasculitis, 7 pyoderma gangrenosum, 7 traumatic ulcers, and 1 neoplastic ulcer. The average area was 16.92 cm2 and the average WBS was 9.17. A total of 16 ulcers (37%) were positive for clinical signs and symptoms of infection and 27 ulcers were negative (63%). Thirty-six ulcers emitted a single fluorescence: cyan (n = 13), red (n = 1), light green (n = 14), and dark red-purple-black (n = 8). Six wounds showed a double fluorescence area: red and cyan (n = 1) and cyan and light green (n = 5). One ulcer emitted a triple fluorescence area: red, cyan, and light green. Overall in 43 ulcers, we found 43 fluorescence and 8 dark red-purple-black color. We found significant data between pH and fluorescence. pH values on wound bed confirm in a noninvasive way the correlation between fluorescence and bacterial burden. Moreover, MolecuLight i:X is able to detect objectively the bacterial proliferation, in contrast with pH which cannot distinguish different types of bacteria.


Subject(s)
Methyl Green , Ulcer , Adult , Male , Female , Humans , Aged , Fluorescence , Bacteria , Hydrogen-Ion Concentration
8.
Environ Res ; 216(Pt 4): 114730, 2023 01 01.
Article in English | MEDLINE | ID: mdl-36372145

ABSTRACT

Owing to the toxicity and difficulty in degradation, how to the effective separation for the residual dyes in the aqueous solution is still an issue with great challenge in the area of environmental protection. Now, to high-efficiency removal of organic dyes from the aqueous solution, we design a unique AlOOH/CoFe2O4 adsorbent with porous CoFe2O4 nanoparticles embedded on the AlOOH fibers using a simple hydrothermal technique and calcination process. The structural properties and surface characteristics of the AlOOH/CoFe2O4 composites are detailedly analyzed by XRD, FTIR, XPS, TEM and SEM. Here, the high SBET and specific porous structure are beneficial to improve the adsorption performance of AlOOH/CoFe2O4 adsorbents. Especially, when the molar ratio of AlOOH to CoFe2O4 in the AlOOH/CoFe2O4 fibers is 1:1, an optimal performance on adsorbing anionic Congo red (CR) and cationic methyl green (MG) dyes can be obtained at pH = 6.29, where the corresponding maximum adsorption capacities reach up to 565.0 and 423.7 mg g-1, respectively. Factors leading to the change in the ability of adsorbing CR and MG dyes are systematically discussed, including contact time, temperature, initial concentrations, and pH values of the solutions. Meanwhile, the uptake of CR and MG dyes can best conform to Langmuir isotherm model and pseudo-second-order adsorption kinetics. The thermodynamic analysis verifies that the dye adsorption process is spontaneous and endothermic. Moreover, from the point view of practical application, the good reusability further makes the as-synthesized magnetic AlOOH/CoFe2O4 composite be a perfect adsorbent with efficiently removing both anionic and cationic dyes from aqueous solutions.


Subject(s)
Nanoparticles , Water Pollutants, Chemical , Congo Red/analysis , Congo Red/chemistry , Adsorption , Coloring Agents/chemistry , Methyl Green , Porosity , Water Pollutants, Chemical/analysis , Hydrogen-Ion Concentration , Kinetics , Nanoparticles/chemistry , Anions/chemistry
9.
Molecules ; 27(23)2022 Dec 01.
Article in English | MEDLINE | ID: mdl-36500484

ABSTRACT

Organic dyes with enduring colors which are malodorous are a significant source of environmental deterioration due to their virulent effects on aquatic life and lethal carcinogenic effects on living organisms. In this study, the adsorption of methyl green (MG), a cationic dye, was achieved by using ZIF-67, which has been deemed an effective adsorbent for the removal of contaminants from wastewater. The characterization of ZIF-67 was done by FTIR, XRD, and SEM analysis. The adsorption mechanism and characteristics were investigated with the help of control batch experiments and theoretical studies. The systematical kinetic studies and isotherms were sanctioned with a pseudo-second-order model and a Langmuir model (R2 = 0.9951), confirming the chemisorption and monolayer interaction process, respectively. The maximum removal capacities of ZIF-67 for MG was 96% at pH = 11 and T = 25 °C. DFT calculations were done to predict the active sites in MG by molecular electrostatic potential (MEP). Furthermore, both Molecular dynamics and Monte Carlo simulations were also used to study the adsorption mechanism.


Subject(s)
Water Pollutants, Chemical , Water Purification , Wastewater , Methyl Green , Kinetics , Water Pollutants, Chemical/chemistry , Water/chemistry , Adsorption , Models, Molecular
10.
Theor Appl Genet ; 135(12): 4277-4288, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36098750

ABSTRACT

KEY MESSAGE: Cp4.1LG15g03420 (CpDsc-1), which encodes a two-component response regulator-like protein (APRR2) in the nucleus, influences dark green stem formation in Cucurbita pepo by regulating the chlorophyll content. Stem color is an important agronomic trait in zucchini (Cucurbita pepo) for robust seeding and high yield. However, the gene controlling the stem color has not been characterized. In this study, we identified a single locus accounting for the dark green stem color of C. pepo (CpDsc-1). Genetic analysis of this trait in segregated populations derived from two parental lines (line 296 with dark green stems and line 274 with light green stems) revealed that stem color was controlled by a single dominant gene (dark green vs. light green). In bulked segregant analysis, CpDsc-1 was mapped to a 2.09-Mb interval on chromosome 15. This region was further narrowed to 65.2 kb using linkage analysis of the F2 population. Sequencing analysis revealed a 14 kb deletion between Cp4.1LG15g03420 and Cp4.1LG15g03360; these two genes both encoded a two-component response regulator-like protein (APRR2). The incomplete structures of the two APRR2 genes and abnormal chloroplasts in line 274 might be the main cause of the light green phenotype. Gene expression pattern analysis showed that only Cp4.1LG15g03420 was upregulated in line 296. Subcellular localization analysis indicated that Cp4.1LG15g03420 was a nuclear gene. Furthermore, a co-dominant marker, G4563 (93% accuracy rate), and a co-segregation marker, Fra3, were established in 111 diverse germplasms; both of these markers were tightly linked with the color trait. This study provided insights into chlorophyll regulation mechanisms and revealed the markers valuable for marker-assisted selection in future zucchini breeding.


Subject(s)
Cucurbita , Cucurbita/genetics , Chromosome Mapping , Methyl Green , Plant Breeding , Genetic Linkage
11.
Anal Methods ; 14(36): 3539-3551, 2022 09 22.
Article in English | MEDLINE | ID: mdl-36018243

ABSTRACT

A simple visual colorimetric method based on arsenomolybdic acid-crystal violet ion-associate pair formation is described for the detection of As in groundwater at about 10, 25 and 50 µg L-1 levels. The pair exhibits light green coloration at ≤5 µg L-1 and blue colorations of distinctly different intensities at about 10, 25 and 50 µg L-1 concentrations of arsenic. High sensitivity is achieved by the preconcentration of As that entails simultaneous sorption of both As(III) and As(V) from groundwater on covellite (CuS) and, later, their elution as As(V), which subsequently participates in the formation of arsenomolybdic acid. The interference in the color development from PO43-ions that are as efficiently sorbed on CuS and eluted as the oxyanions of As is eliminated by their selective removal by Ce4+ ions under basic (pH ∼ 8.5) conditions. The removal is caused by the formation of cerium phosphate and its co-precipitation with calcium hydroxide. SiO42- ions do not interfere in the process as they are not sorbed by CuS. Groundwater containing ≤0.5 mg L-1 P and ≥200 mg L-1 total dissolved solid can be conveniently analysed by the method. The direct sensing of As(III) as well as As(V), the use of benign and easily available chemicals, the absence of any hazardous by-product, undiminished applicability in sunlight, the testing procedure lasting only for about 30 min, and rapidity are the major advantages of the method. Thus, the method is potentially well-suited for the on-site testing of groundwater potability under different regulations.


Subject(s)
Arsenic , Groundwater , Water Pollutants, Chemical , Arsenic/analysis , Arsenic/chemistry , Calcium Hydroxide/analysis , Copper , Gentian Violet , Groundwater/chemistry , Ions , Methyl Green , Water Pollutants, Chemical/analysis
12.
Chemosphere ; 308(Pt 1): 136212, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36041524

ABSTRACT

Two-dimensional (2D) materials have fascinated the researchers to exploit their properties including large surface area, ability to act as a support and to form face-to-face interfacial contact with other 2D materials for fabricating efficient photocatalytic materials. In this work, Bi2WO6, TiO2 and Ti3C2 nanosheets have been used synthesizing different series of binary Bi2WO6-TiO2 and ternary Bi2WO6-TiO2-Ti3C2 2D nanocomposites by an electrostatic self-assembly synthesis route. The as-prepared pristine materials and binary and ternary nanocomposites were characterized by different structural, morphological and compositional characterization techniques to confirm their successful synthesis and 2D morphology. It was found that the optimized Bi2WO6-TiO2 (20 wt%) and Bi2WO6-TiO2 (20 wt%)-Ti3C2 (5 wt%) nanocomposites showed 97.0% and 98.5% degradation of methyl green in 80 min and 40 min, respectively, which was higher than their pristine counterparts. The enhanced activity was credited to the large surface area offered by 2D nanocomposites, pollutant adsorption and enhanced photogenerated charge separation and transfer facilitated by S-scheme mechanism and face-to-face interfacial contact of different components of these nanocomposites. This work delivers an example of highly efficient 2D nanocomposites and discusses the role of Ti3C2 as an electron acceptor in S-scheme photocatalytic system.


Subject(s)
Environmental Pollutants , Nanocomposites , Methyl Green , Nanocomposites/chemistry , Sunlight , Titanium/chemistry
13.
Water Sci Technol ; 85(4): 1040-1052, 2022 Feb.
Article in English | MEDLINE | ID: mdl-35228352

ABSTRACT

Cadmium sulphide (CdS) nanoparticles (NPs) were synthesized through hydrothermal route and characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), Energy dispersive X-ray analysis, Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy and Thermo gravimetric analysis (TGA).The band gap of CdS nanoparticles was found to be 2.38 eV. CdS NPs are crystalline aggregates with hexagonal structure as shown by SEM and XRD analysis. TGA study revealed that the synthesized nanomaterials were very stable to temperature and only 6.54% total loss occurred during heating range (25 °C-600 °C).The CdS NPs were used for the first time against the degradation of Eosin B (EB) and Methyl green (MG) dyes in aqueous solution.The degradation of EB and MG over CdS nanocatalysts followed second order kinetics. The predicted activation energies for both the dyes' reactions were 61.1 kJ/mol and 32.11 kJ/mol, respectively. About 95% and 90% dye degradation was observed at the time interval of 160 minutes for EB and MG, respectively. High percent degradation of EB was observed at high pH (pH 0) while at low pH (pH 4) high percent degradation was found for MG dye. Maximum dye degradation was found at the optimal dose (0.03 g/L) of the catalyst and at low dye concentration. The rate of EB and MG dye degradation was found to increase with increase in temperature up to 45 °C. The recyclability study showed that CdS nanoparticles could be reused for the degradation of the given dyes. Good antibacterial activity against Staphylococcus aureus was shown by CdS NPs. From the biocompatibility it was confirmed that CdS NPS are bioincompatible compatible.


Subject(s)
Coloring Agents , Methyl Green , Cadmium Compounds , Catalysis , Coloring Agents/chemistry , Eosine I Bluish , Kinetics , Sulfides , Thermodynamics , X-Ray Diffraction
14.
Environ Sci Pollut Res Int ; 29(53): 79995-80004, 2022 Nov.
Article in English | MEDLINE | ID: mdl-35199267

ABSTRACT

In this study, a simple, one-pot, and eco-friendly biosynthesis of silver nanoparticles (AgNPs) was accomplished with the use of aqueous leaves extract of Cestrum nocturnum L.(AECN). Different techniques like ultraviolet-visible (UV-Vis) spectrophotometry, Fourier transform infrared (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning area electron diffraction were used to investigate the optical, operational, and physical properties of the green synthesized AECN-AgNPs.The AECN-AgNPs were further used for the detection of Hg2+ by UV-Vis and electrochemical methods. The disintegration of the AECN-AgNPs solution caused the formation of an Ag-Hg amalgam, which caused discoloration of the solution. Sensing performance for a variety of metals such as Na+, K+, Mg2+, Ca2+, Ni2+, Cu 2+, Fe3+, Zn2+, Co2+, Cd2+, Pb2+, As3+, and Mn2+ at 10-mM concentrations was measured in order to determine the selectivity of the sensor towards the Hg2+. For the electrochemical determination of 2 + Hg2+ , AECN-AgNPs were immobilized on a glassy carbon (GC) electrode, and the resulting modified electrode (GC/AECN-AgNPs) was characterized by cyclic voltammetry. This phenomenon is advantageously used for the sensitive determination of trace level Hg2+. GC/AECN-AgNPs demonstrated a linear calibration range of 100 nM to 10 µM and a limit of detection of 21 nM for Hg2+ determination.


Subject(s)
Cestrum , Mercury , Metal Nanoparticles , Silver/chemistry , Metal Nanoparticles/chemistry , Methyl Green , Cadmium , Lead , Water/chemistry , X-Ray Diffraction , Plant Extracts/pharmacology , Carbon , Spectroscopy, Fourier Transform Infrared
15.
Arch Pathol Lab Med ; 146(10): 1268-1272, 2022 10 01.
Article in English | MEDLINE | ID: mdl-35104313

ABSTRACT

CONTEXT.­: There is no standardized process for utilization of periodic acid-Schiff during intraoperative frozen sections to identify fungal organisms. OBJECTIVE.­: To develop a rapid staining process for fresh tissue with periodic acid-Schiff during intraoperative consultation and develop an appropriate control block. DESIGN.­: Muscle tissue was inoculated with 2 species of fungus (Aspergillus fumigatus and Paecilomyces spp) and grown at 3 different temperatures for 72 hours. Inoculated tissue was embedded in optimal cutting temperature compound, cut, and stained using a modified periodic acid-Schiff stain. The optimal control was determined for future use as the standard control. Multiple control slides were cut and stained, using successively shorter time intervals for each step. The staining process that provided accurate results in the shortest amount of time was deemed ultra-rapid periodic acid-Schiff. This method was validated by carryover studies and clinical specimens. RESULTS.­: Paecilomyces spp incubated at 30°C for 72 hours was the most optimal positive control, with numerous yeast and hyphal forms. The fastest staining process involved 2 minutes of periodic acid and Schiff reagent and 10 dips of light green solution. Tap water was as effective as distilled water. Validation was successfully achieved. Clinical cases all stained identical to formalin-fixed, paraffin-embedded tissue stained with hematoxylin-eosin and periodic acid-Schiff. CONCLUSIONS.­: Ultra-rapid periodic acid-Schiff provides fast and reliable identification of fungal organisms on fresh tissue. Development of a concurrent positive control allows for quality control and validation.


Subject(s)
Frozen Sections , Methyl Green , Coloring Agents , Eosine Yellowish-(YS) , Formaldehyde , Fungi , Hematoxylin , Humans , Periodic Acid , Staining and Labeling , Water
16.
Int Microbiol ; 25(4): 669-678, 2022 Nov.
Article in English | MEDLINE | ID: mdl-33835333

ABSTRACT

Colpodella spp. are free-living flagellates closely related to the apicomplexans. Human infections by Colpodella sp. have been reported. A biflagellated trophozoite and cyst stage comprise the known life cycle stages of Colpodella sp. However, the process of encystation and excystation within the life cycle is unclear. Life cycle stages initiating human infections are unknown. We performed a detailed investigation of the life cycle of Colpodella sp. (ATCC 50594) in culture using Sam-Yellowe's trichrome stains and differential interference contrast (DIC) for light microscopy and fluorescence microscopy of Congo red-stained cells and investigated ultrastructure using transmission electron microscopy (TEM). We report previously undocumented stages of Colpodella sp. Asymmetric and asynchronous division was detected inside cysts by trichrome staining and by TEM. Odd-numbered juveniles and cysts containing more than four juvenile trophozoites were identified. Live imaging of active cultures captured the excystation and egress of juvenile trophozoites and confirmed the presence of multinucleate cysts. The ultrastructure of the multinucleate cyst is reminiscent of apicomplexan schizonts. Insights gained from the life cycle stages observed in culture allowed the construction of the life cycle of Colpodella sp. Knowledge of the life cycle will aid biochemical and molecular characterization of Colpodella sp. and help identify stages in human infections.


Subject(s)
Apicomplexa , Congo Red , Animals , Azo Compounds , Eosine Yellowish-(YS) , Humans , Life Cycle Stages , Methyl Green , Microscopy, Electron , Trophozoites
17.
Am J Surg Pathol ; 46(2): 200-212, 2022 02 01.
Article in English | MEDLINE | ID: mdl-34411028

ABSTRACT

Venous invasion (VI) is a powerful yet underreported prognostic factor in colorectal cancer (CRC). Its detection can be improved with an elastin stain. We evaluated the impact of routine elastin staining on VI detection in resected CRC and its relationship with oncologic outcomes. Pathology reports from the year before (n=145) and the year following (n=128) the implementation of routine elastin staining at our institution were reviewed for established prognostic factors, including VI. A second review, using elastin stains, documented the presence/absence, location, number, and size of VI foci. The relationship between VI and oncologic outcomes was evaluated for original and review assessments. VI detection rates increased from 21% to 45% following implementation of routine elastin staining (odds ratio [OR]=3.1; 95% confidence interval [CI]: 1.8-5.3; P<0.0001). The second review revealed a lower VI miss rate postimplementation than preimplementation (22% vs. 48%, respectively; P=0.007); this difference was even greater for extramural VI-positive cases (9% vs. 38%, respectively; P=0.0003). Missed VI cases postimplementation had fewer VI foci per missed case (P=0.02) and a trend towards less extramural VI than those missed preimplementation. VI assessed with an elastin stain was significantly associated with recurrence-free survival (P=0.003), and cancer-specific survival (P=0.01) in contrast to VI assessed on hematoxylin and eosin alone (P=0.053 and 0.1, respectively). The association between VI and hematogenous metastasis was far stronger for elastin-detected VI (OR=11.5; 95% CI: 3.4-37.1; P<0.0001) than for hematoxylin and eosin-detected VI (OR=3.7; 95% CI: 1.4-9.9; P=0.01). Routine elastin staining enhances VI detection and its ability to stratify risk in CRC and should be considered for evaluation of CRC resection specimens.


Subject(s)
Colorectal Neoplasms/chemistry , Elastin/analysis , Veins/chemistry , Adult , Aged , Aged, 80 and over , Azo Compounds , Biomarkers, Tumor , Biopsy , Colectomy , Colorectal Neoplasms/mortality , Colorectal Neoplasms/pathology , Colorectal Neoplasms/surgery , Coloring Agents , Eosine Yellowish-(YS) , Female , Humans , Male , Methyl Green , Middle Aged , Neoplasm Invasiveness , Predictive Value of Tests , Risk Assessment , Risk Factors , Staining and Labeling , Treatment Outcome , Veins/pathology , Young Adult
18.
Int J Biol Macromol ; 192: 1-6, 2021 Dec 01.
Article in English | MEDLINE | ID: mdl-34619269

ABSTRACT

New chitosan Schiff base (CS-NB) and its CS-NB-NiFe nanocomposite have been prepared and characterized by FTIR spectroscopy, XRD, SEM and DSC. FT-IR spectra and XRD patterns revealed the preparation of chitosan Schiff base CS-NB and its CS-NB-NiFe nanocomposite. DSC demonstrated the endo and exothermic correspondence the evaporation of solvent and decomposition of pyranose ring, respectively. Antibacterial activities was evaluated for the as-prepared compounds against two Gram-positive (Staphylococcus aureus and Bacillus cereus) and two Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria and the results shows that the antibacterial activities of the compounds are found to be stronger than that of chitosan. The order of antibacterial effect according to inhibitory zone around is as follows: S. aureus > E. coli > B. cereus > P. aeruginosa. In addition, the removal of methyl green (MG) dye using CS-NB and its CS-NB-NiFe nanocomposite were analyzed and results showed that the compounds can be effectively used to remove of MG from aqueous solution. Results show that the percentage removal of MG by nanocomposite is higher than Schiff base.


Subject(s)
Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Chitosan/chemistry , Methyl Green/chemistry , Nanocomposites/chemistry , Schiff Bases/chemistry , Adsorption , Chemical Phenomena , Kinetics , Microbial Sensitivity Tests , Molecular Structure , Nanocomposites/ultrastructure , Nanoparticles , Spectroscopy, Fourier Transform Infrared , X-Ray Diffraction
19.
Am J Surg Pathol ; 45(10): 1419-1427, 2021 10 01.
Article in English | MEDLINE | ID: mdl-33756495

ABSTRACT

Venous invasion (VI) is a powerful prognostic factor in colorectal cancer (CRC) that is widely underreported. The ability of elastin stains to improve VI detection is now recognized in several international CRC pathology protocols. However, concerns related to the cost and time required to perform and evaluate these stains in addition to routine hematoxylin and eosin (H&E) stains remains a barrier to their wider use. We therefore sought to determine whether an elastin trichrome (ET) stain could be used as a "stand-alone" stain in CRC resections, by comparing the sensitivity, accuracy, and reproducibility of detection of CAP-mandated prognostic factors using ET and H&E stains. Representative H&E- and ET-stained slides from 50 CRC resections, including a representative mix of stages and prognostic factors, were used to generate 2 study sets. Each case was represented by H&E slides in 1 study set and by corresponding ET slides from the same blocks in the other study set. Ten observers (3 academic gastrointestinal [GI] pathologists, 4 community pathologists, 3 fellows) evaluated each study set for CAP-mandated prognostic factors. ET outperformed H&E in the assessment of VI with respect to detection rates (50% vs. 28.6%; P<0.0001), accuracy (82% vs. 59%, P<0.0001), and reproducibility (k=0.554 vs. 0.394). No significant differences between ET and H&E were observed for other features evaluated. In a poststudy survey, most observers considered the ease and speed of assessment at least equivalent for ET and H&E for most prognostic factors, and felt that ET would be feasible as a stand-alone stain in practice. If validated by others, our findings support the use of ET, rather than H&E, as the primary stain for the evaluation of CRC resections.


Subject(s)
Azo Compounds , Biomarkers, Tumor/analysis , Colorectal Neoplasms/chemistry , Coloring Agents , Elastin/analysis , Eosine Yellowish-(YS) , Methyl Green , Staining and Labeling , Veins/chemistry , Colorectal Neoplasms/pathology , Colorectal Neoplasms/surgery , Feasibility Studies , Humans , Neoplasm Invasiveness , Observer Variation , Predictive Value of Tests , Reproducibility of Results , Veins/pathology
20.
Mod Pathol ; 34(4): 808-822, 2021 04.
Article in English | MEDLINE | ID: mdl-33299110

ABSTRACT

Non-alcoholic steatohepatitis (NASH) is a fatty liver disease characterized by accumulation of fat in hepatocytes with concurrent inflammation and is associated with morbidity, cirrhosis and liver failure. After extraction of a liver core biopsy, tissue sections are stained with hematoxylin and eosin (H&E) to grade NASH activity, and stained with trichrome to stage fibrosis. Methods to computationally transform one stain into another on digital whole slide images (WSI) can lessen the need for additional physical staining besides H&E, reducing personnel, equipment, and time costs. Generative adversarial networks (GAN) have shown promise for virtual staining of tissue. We conducted a large-scale validation study of the viability of GANs for H&E to trichrome conversion on WSI (n = 574). Pathologists were largely unable to distinguish real images from virtual/synthetic images given a set of twelve Turing Tests. We report high correlation between staging of real and virtual stains ([Formula: see text]; 95% CI: 0.84-0.88). Stages assigned to both virtual and real stains correlated similarly with a number of clinical biomarkers and progression to End Stage Liver Disease (Hazard Ratio HR = 2.06, 95% CI: 1.36-3.12, p < 0.001 for real stains; HR = 2.02, 95% CI: 1.40-2.92, p < 0.001 for virtual stains). Our results demonstrate that virtual trichrome technologies may offer a software solution that can be employed in the clinical setting as a diagnostic decision aid.


Subject(s)
Azo Compounds , Coloring Agents , Eosine Yellowish-(YS) , Image Interpretation, Computer-Assisted , Liver Cirrhosis/diagnosis , Liver/pathology , Methyl Green , Microscopy , Neural Networks, Computer , Non-alcoholic Fatty Liver Disease/diagnosis , Staining and Labeling , Adolescent , Adult , Aged , Aged, 80 and over , Biopsy , Child , Clinical Decision-Making , Decision Support Techniques , Female , Hematoxylin , Humans , Liver Cirrhosis/pathology , Male , Middle Aged , Non-alcoholic Fatty Liver Disease/pathology , Predictive Value of Tests , Reproducibility of Results , Retrospective Studies , Severity of Illness Index , Software , Young Adult
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