• Journal of Veterinary Science
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• v.19 no.6
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• pp.744-749
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• 2018

Dapsone, an antibiotic, has been used to cure leprosy. It has been reported that dapsone has anti-inflammatory activity in hosts; however, the anti-inflammatory mechanism of dapsone has not been fully elucidated. The present study investigated the anti-inflammatory effects of dapsone on bone marrow cells (BMs), especially upon exposure to lipopolysaccharide (LPS). We treated BMs with LPS and dapsone, and the treated cells underwent cellular activity assay, flow cytometry analysis, cytokine production assessment, and reactive oxygen species assay. LPS distinctly activated BMs with several characteristics including high cellular activity, granulocyte changes, and tumor necrosis factor alpha ($TNF-{\alpha}$) production increases. Interestingly, dapsone modulated the inflammatory cells, including granulocytes in LPS-treated BMs, by inducing cell death. While the percentage of Gr-1 positive cells was 57% in control cells, LPS increased that to 75%, and LPS plus dapsone decreased it to 64%. Furthermore, dapsone decreased the mitochondrial membrane potential of LPS-treated BMs. At a low concentration ($25{\mu}g/mL$), dapsone significantly decreased the production of $TNF-{\alpha}$ in LPS-treated BMs by 54%. This study confirmed that dapsone has anti-inflammatory effects on LPS-mediated inflammation via modulation of the number and function of inflammatory cells, providing new and useful information for clinicians and researchers.

• Biomedical Science Letters
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• v.1 no.1
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• pp.19-25
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• 1995

Characterization of immune cell subpopulations in the bovine was performed using a direct immunofluorescence technique adaptable for routine and repeated monitoring. This whole blood procedure is faster and requires less volume than conventional density gradient isolation methods. Low inter-and intra-animal variations were seen in hematology parameters and in CD4, CD8 and CD2 lymphocyte subtypes. CD4 values were 30% of lymphocytes in male and 32% in females. Thriteen percent were CD8 in males and 13% in females. CD4: CD8 ratios were approximately two in both sexes. Fiffty three percent were CD2 in males and 54% in females. The mean RBC counts of peripheral blood were 7.20$\times10^6/{mm}^3$ for male cattle and 6.36$\times10^6/{mm}^3$ for females. The mean WBC counts were 8.09$\times10^3/{mm}^3$ for males and 7.09$\times10^3/{mm}^3$ for females. The percent of lymphocytes(63-65%) was higher than the percent of neutrophils(17-18%), the percent of eosinophils(11-15%), te percent of monocytes(4-5%), and percent of basophiles(1%).

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.4
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• pp.801-809
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• 2002

CSBHT is known to improve immunological response in mice and humans. In this study, CSBHT effect was examined in the context of CD4+ T cells' survival and TCR/CD3 induced activation responses. Spleen cells from 8 week BALB/c mice were cultured in CSBHT containing medium without activation for 24, 48 hr. The MTS assay and revealed that CSBHT did not stimulate spleen lymphocytes as mitogen. Spleen lymphocytes were treated with anti-CD3e/anti-CD28 antibodies for 48hr. Flow cytometry revealed that activity of T cell decreased with CSBHT concentration. CD4+ T cells were isolated and cultured ,in CSBHT containing medium for 48 hr. CSBHT did not affect survival of sorted CD4+ T cells without any involvement of APC. In order to evaluate the direct effect of CSBHT on helper T cells's proliferative capacity prior to activation, CD4+ T cells are isolated after 24hr of culture in CSBHT containing medium and activated with and without anti-CD3e/anti-CD28 activation for 48hr. A higher level of CD69 was observed in 1 ㎍/㎖ of CSBHT treatment than control using flow cytometry. But low CD69 expression was observed in 5㎍/㎖ of CSBHT treatment. Expression of mRNA for cytokines in CD4+ T cell revealed that IL-2 expression was increased in 1 ㎍/㎖. The expression of IL-2R α, INF- γ were increased with concentration. On the other hand mRNA of IL-4 was decreased in dose dependent manner. Results suggest that CSBHT may be desirable for CD4+ T cell's activity in immune responses. Further more, CSBHT may relatively activate Th1 and inactivate Th2.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.8
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• pp.3727-3731
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• 2016

Background: The most common type of ocular lymphoma is non-Hodgkin lymphoma (NHL), categorized into two groups: indolent (slow growing) and aggressive (rapid growing). Differentiating benign reactive lymphoid hyperplasia (RLH) from malignant ocular adnexal lymphoma (OAL) is challenging. Histopathology, immunohistochemistry (IHC) and flow cytometry have been used as diagnostic tools in such cases. Materials and Methods: In this retrospective case series, from 2002 to 2013 at Farabi Eye Center, 110 patients with ocular lymphoproliferative disease were enrolled. Prevalence, anatomical locations, mean age at diagnosis and the final diagnosis of the disease with IHC were assessed. Comparison between previous pathologic diagnoses and results of IHC was made. Immunoglobulin light chains and B-cell and T-cell markers and other immuno-phenotyping markers including CD20, CD3, CD5, CD23, CD10, CYCLIND1 and BCL2 were evaluated to determine the most accurate diagnosis. The lymphomas were categorized based on revised European-American lymphoma (REAL) classification. Results: Mean age ${\pm}$ SD (years) of the patients was $55.6{\pm}19.3$ and 61% were male. Patients with follicular lymphoma, large B-cell lymphoma or chronic lymphocytic leukemia/small cell lymphoma (CLL/SLL) tended to be older. Nine patients with previous diagnoses of low grade B-cell lymphoma were re-evaluated by IHC and the new diagnoses were as follows: extranodal marginal zone lymphoma(EMZL) (n=1), SLL(n=1), mantle cell lymphoma (MCL) (n=3), reactive lymphoid hyperplasia RLH (n=2). Two cases were excluded due to poor blocks. Flow cytometry reports in these seven patients revealed SLL with positive CD5 and CD23, MCLwith positive CD5 and CyclinD1 and negative CD23, EMZL with negative CD5,CD23 and CD10. One RLH patient was negative for Kappa/Lambda and positive for CD3 and CD20 and the other was positive for all of the light chains, CD3 and CD20. Orbit (49.1%), conjunctiva (16.1%) and lacrimal glands (16.1%) were the most common sites of involvement. Conclusions: Accurate pathological classification of lesions is crucial to determine proper therapeutic approaches. This can be achieved through precise histologic and IHC analyses by expert pathologists.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.1
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• pp.215-217
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• 2016

Background: Chronic lymphocytic leukemia (CLL) is the most frequent form of leukemia in Europe and North America, and it mainly affects older individuals. Many approaches are implemented in the management of CLL from a watch and wait (W&W) strategy to chemotherapeutic regimens. We here reviewed our clinical practice for the relevance of the W&W strategy in Lebanese CLL patients. Materials and Methods: A total of 95 patients with CLL diagnosed in four institutions in Lebanon, between 1992 and 2013, were selected and their files were reviewed. Characteristics of these patients were noted including age, sex, RAI and Binet scores, CBC values, presence of hepatomegaly or splenomegaly, performance of bone marrow biopsy or peripheral blood flux cytometry for diagnosis, adoption of W&W strategy, different chemotherapeutic regimens and the indications for treatment. Results: Some 38 patients (40%) diagnosed with CLL were women and 57 (60%) were men with a mean age of 65.1 years [36-89]. Of the total, 50.5%, 17.2%, 14%, 7.5% and 10.8% had an RAI score at diagnosis of 0, 1, 2, 3 and 4, respectively, while 65.6%, 17.2% and 17.2% had Binet scores of I, II and III. The mean lymphocyte count at diagnosis was $39885/mm^3$ [1596-290000], the mean hemoglobin level was 12.7 g/dl [6.2-17] and the mean platelet count was $191255/mm^3$ [14000-458000]. While 26.3% of patients with CLL had splenomegaly, only 7.4% had hepatomegaly. Some 33.7% had undergone a bone marrow biopsy, 66.3% flow cytometry of circulating blood and 5.3% a lymph node biopsy. Overall, the W&W was adopted in 62.4% (58) of patients with a mean duration of 37.7 months [3-216]. The W&W was used in 82.6%, 73.3%, 46.2%, 14.3% and 0% of patients having RAI scores of 0, 1, 2, 3 and 4, respectively, and, it was used in 80%, 46.7% and 6.25% with Binet scores of I, II and III. The most frequent indication for treatment was anemia and thrombocytopenia, accounting for 32.7% of cases. The most frequently used chemotherapeutical regimens were chlorambucil until the end of the last century and flufarabine-cyclophosphamide-rituximab during the last decade. Conclusions: This retrospective review of CLL clinical practice showed an important implementation of the W&W strategy with a long duration, especially in early stage cases with low RAI or Binet scores.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.382-389
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• 2017

Currently, reticulocyte experimental calculation technology used in clinical laboratories are divided two types: manual and automated. Manual reticulocyte counting using a microscopy lacks accuracy due particularly to its low reproducibility, affecting the accuracy of manual reticulocyte count. Moreover, Automatic blood corpuscle analyzer flow cytometry is difficult to be used in underdeveloped countries and small scale laboratories due to relatively high cost. Therefore, this study tried to find a new method to complement these drawbacks. The aim of this study was to compare the stained reticulocytes count by spectrophotometer and also to analyze the statistics of spectrophotometer and flow cytometer. The same 8 EDTA samples were repeated 36 times to compare the agreement between spectrophotometer and flow cytometer. This study measured the specimen diluted 600 times at 700~780 nm by 10 differences. Wavelength between 710 to 730 by absorbance showed a positive correlation between standard data and test data (r=0.967, p<0.01), presenting a correlation between variables. Statistical analyses of regression for test and standard parametric data, the optimal dilution factor was 600 times. Therefore, this study tried to technical utilizes such as contributing economical for the reticulocyte absorbance apply from the auto spectrophotometer, a monitoring system for the reticulocyte relation anemia, etc. Therefore, more extensive studies, including an auto chemical analyzer application, will be needed.

• Korean Journal of Food Science and Technology
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• v.48 no.5
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• pp.454-460
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• 2016

In this study, we investigated the possibility of Droplet digital PCR (ddPCR) for detection of foodborne pathogens. ddPCR combines partitioning of PCR reactions into several thousands or millions of individual droplets in a water-oil emulsion, and counting of positive PCR reaction using flow cytometry. Four species of foodborne pathogens, Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli O157:H7, were used to quantify the target sequence with each of the designed primers and double stranded DNA-binding Evagreen dye. All tested foodborne pathogens showed a detection limit ranging from $100fg/{\mu}L$ to $10ng/{\mu}L$. It was concluded that ddPCR could be used to detect very low concentrations of foodborne pathogens from complex food matrices. For multi-detection of target pathogens, we also tested the samples using multiplex ddPCR and obtained successful results.

• The Journal of the Korean bone and joint tumor society
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• v.5 no.1
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• pp.70-75
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• 1999

We present a case of parosteal osteosarcoma of the femur with secondary dedifferentiation. This 57-year-old woman was radiologically diagnosed as a parosteal osteosarcoma in 1987. In 1992, excisional biopsy revealed a classical parosteal osteosarcoma with diploidy DNA pattern. In 1998, she revisited due to a recurrent tumor with pathologic fracture. The resected specimen showed a classic parosteal osteosarcoma with area of dedifferentiation, showing high-grade spindle cell sarcoma. This dedifferentiated area revealed aneuploidy cell population on DNA flow cytometry. This case reminds us that not all parosteal osteosarcomas are low-grade lesions. Some low-grade lesions may dedifferentiate to become high-grade tumors after inadequate excision.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.12
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• pp.4849-4852
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• 2015

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been investigated as an effective agent to treat various cancers. Cancer stem cells are resistant to TRAIL treatment, but the mechanism of TRAIL resistance remains unknown. In this study, brain cancer stem cells were isolated by CD133 magnetic sorting, and the number of CD133 positive cells detected by flow cytometry. The self-renewing capacity of brain cancer stem cells was examined by a neurosphere formation assay, and the percentage of cell death after TRAIL treatment was examined by an MTS assay. Expression of DR5, FADD, caspase 8 and BCL2 proteins was detected by western blot. The amount of CD133 positive cells was enriched to 71% after CD133 magnetic sorting. Brain cancer stem cell neurosphere formation was significantly increased after TRAIL treatment. TRAIL treatment also reduced the amount of viable cells and this decrease was inhibited by a caspase 8 inhibitor or by the pan-caspase inhibitor z-VAD (P<0.05). Brain cancer stem cells expressed lower levels caspase 8 protein and higher levels of BCL2 protein when compared with CD133 negative cells (P<0.05). Our data suggest that TRAIL resistance is related to overexpression of BCL2 and low expression of caspase 8 which limit activation of caspase 8 in brain cancer stem cells.

• Biomedical Science Letters
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• v.7 no.4
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• pp.181-190
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• 2001

This study was designed to investigate the effects of reactive oxygen species (ROS) on DNA stability in human spermatozoa. To verify human spermatozoa were incubated with xanthine-xanthine oxidase (X 100$\mu$M-XO 50 mlU ~ 400 mIU), $H_2O_2$ (125 $\mu$M ~ 1 mM), sodium nitroprusside (SNP 0.1 $\mu$M ~ 100 $\mu$M) or lymphocyte. Otherwise, spermatozoa were incubated under low $O_2$ (5%) condition. Damage of sperm DNA was analyzed by single cell electrophoresis (Comet assay) and flow cytometry after acridine orange staining. In the presence of ROS, there was increase in DNA damage. The rate of DNA single strand breakage (9.0$\pm$1.0% ~ 46.0$\pm$4.6%) and DNA fragmentation (7.51$\pm$1.0% ~ 29.5$\pm$4.6%) were similar regardless of the kinds of ROS and exposure time. DNA damage in the lower $O_2$ condition (5%) was lower than ambient $O_2$ condition (20%). Taken together, it suggested that sperm DNA might be damaged by ROS. In the presence of ROS, increase in DNA damage and chromatin instability was obvious in spite of short exposure. Although present study reconfirmed that sperm incubation in the low concentration of ROS have the benefit m the induction of capacitation and Ah, the increase in DNA damage by ROS and possible genetic problem should be considered before the human trials.