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Constitutive Host Resistance
Published in Julius P. Kreier, Infection, Resistance, and Immunity, 2022
The polymorphonuclear leukocytes are also known as granulocytes because of the many granules found in their cytoplasm. The contents of the granules determine the cell′s staining properties and help to distinguish the different ceils of the granulocytic series. Three types of granulocytes based on staining characteristics have been described. These are neutrophils, eosinophils, and basophils. The stain most commonly used to distinguish the ceils is the Giemsa stain, an azure B eosinate. It is prepared by interaction between two basic dyes, azure B and methylene blue, and an acidic dye, eosin. The basophilic, or cationic, components react with the negatively charged molecules of the granules, staining them blue. The acidophilic, or anionic, component reacts with the positively charged molecules, staining them red. The granules of the neutrophil are not ionically charged at physiological pHs and thus do not stain; they assume a grey or neutral tint.
Understanding Microbiology Culture Results
Published in Firza Alexander Gronthoud, Practical Clinical Microbiology and Infectious Diseases, 2020
Intestinal parasites can be detected in stool using a direct wet preparation or an iodine stain (i.e. Giardia lamblia). With microscopy, trophozoites can be detected in the stool if either the stool is processed within 1 hour (‘hot stool’) or if the stool is fixated. A modified Ziehl–Neelsen stain can also be used to detect Cryptosporidium oocysts. A Giemsa stain is commonly used for microscopy of sterile samples, i.e. malaria parasites in blood. For moulds including Aspergillus spp., a lactophenol cotton blue stain or calcofluor-white stain is commonly used. India ink is used for Cryptococcus spp. in CSF. A rapid method to diagnose dermatophytes is a KOH test on skin scrapings (i.e. dermatophytes). Microscopy can also be performed on histology tissue sections with various stains including periodic acid-Schiff stain or a Grocott silver stain.
Pneumocystis carinii
Published in Peter D. Walzer, Robert M. Genta, Parasitic Infections in the Compromised Host, 2020
Peter D. Walzer, C. Kurtis Kim, Melanie T. Cushion
The second type of P. carinii stains stain trophozoites, intracystic bodies, and all intermediate forms. Giemsa stain is the prototype, but similar results can be obtained with Wright-Giemsa and polychrome methylene blue (305,310, 311). We favor Diff Quik, a variant of the Wright-Giemsa stain, because it is very rapid (can be accomplished in less than 1 min) and gives staining results similar to those achieved with the standard Giemsa stain (137,261). Recent studies indicate that P. carinii can also be seen with Wright's and Gram's stains in lung imprint smears and bronchoalveolar lavage fluid (311-316), but it is unclear what specific role these stains will have in diagnosis of this infection.
Retinoic acid (all-trans) presents antioxidant properties within human ovary and reduces progesterone production by human granulosa cells
Published in Systems Biology in Reproductive Medicine, 2023
Bruno M. Fonseca, Rebeca Cruz, Beatriz Pinto, Lia Costa, Eduarda Felgueira, Pedro Oliveira, Susana Casal, Irene Rebelo
For cell viability determination, COV434 and hGC were seeded in 96-well plates at a density of 3.5 × 105 and 7.0 × 105, respectively. After 24 h, cells were exposed to either all-trans-retinoic acid (atRA) or α-tocopherol (0.01–50 µM) for 24, 48 and 72 h and the tetrazolium-based MTT assay was used to determine cell viability. For morphology evaluation, cells were exposed to either atRA or α-tocopherol for 48 h in 24-well plates. For Giemsa and Höechst staining, cells were fixed with a 4% paraformaldehyde solution. Giemsa stain was added to the cells for 30 min, then cells were observed by light microscopy. For Höechst and Acridine Orange staining, cells were incubated with 0.5 μg/ml Höechst 33342 or 5 μg/mL Acridine Orange (AO) at 37°C for 20 min and were observed by fluorescence microscopy (Eclipse Ci, Nikon, Japan).
Macrophage targeting with the novel carbopol-based miltefosine-loaded transfersomal gel for the treatment of cutaneous leishmaniasis: in vitro and in vivo analyses
Published in Drug Development and Industrial Pharmacy, 2021
Sibgha Batool, Fatima Zahid, Fakhar- Ud-Din, Syeda Sohaila Naz, Muhammad Junaid Dar, Muhammad Waseem Khan, Alam Zeb, Gul Majid Khan
HePC solution and HePCT were evaluated for their anti-leishmanial activity using intramacrophage amastigote model [16]. HePC solution was used a positive control. Macrophage cells were taken in 24-well culture plate with coverslips (2 × 104 cells/well), incubated for 24 h in 5% CO2 incubator and allowed to fix. Fixed cells were infected with promastigotes of Leishmania major at a ratio of 10:1 (promastigotes: macrophages). Infected cells were incubated at 37 °C for 24 h followed by washing to remove free promastigotes. HePC solution and HePCT were added in various quantities (100, 50, 25, 12.5, 6.25, 3.125 µgmL−1) into the plate followed by incubation for 24 h. Subsequently, staining with 10% Giemsa stain was done and observed under microscope. The % inhibition and selectivity index (SI) were calculated by using the following equations and half maximal inhibitory concentration (IC50) was determined using regression analysis [16].
Protective Effect of Chrysin, a Dietary Flavone against Genotoxic and Oxidative Damage Induced by Mitomycin C in Balb/C Mice
Published in Nutrition and Cancer, 2021
Aïcha Sassi, Jihed Boubaker, Amira Loussaief, Khaoula Jomaa, Kamel Ghedira, Leila Chekir-Ghedira
The CA assay was carried out as described by Soumaya et al. (28) with minor modifications. The animals within the different treatment groups received a single intraperitoneal (i.p) injection of the tested substance solution (MMC, chrysin or corn oil). Then vinblastine (200 μl; 250 μg/ml) was injected into the animals 45 min before sacrifice in order to block dividing cells in metaphasis. Bone marrow cells were collected by flushing with 0.56% KCl (pre-warmed at 37 °C) from the femur and tibia of mice and incubated for 20 min at 37 °C. The material was centrifuged at 1500 rpm for 5 min, fixed in freshly prepared aceto-methanol buffer (acetic acid and methanol in the ratio 1:3, v/v) followed by refrigeration for 30 min. The material was centrifuged and re-suspended in aceto-methanol buffer. The slides were prepared by dropping the sample on chilled slides and run over the flame. Staining was done in 5% buffered Giemsa stain (pH 7.0) for 20 min. Then the slides were air-dried for 24 h and covered with cover slips. After coding the slides, the chromosomes of 100 cells blocked in metaphase were examined for chromosome abnormalities at a magnification of 100 × using an optical microscope (Olympus, France). Three replicates (300 metaphases per dose level) for each control and treated group were conducted.