Monocyclicity in the Function of the Erythroid Hematopoietic Lineage in Teleost Fish Exemplified by Platichthys Flesus (Linnaeus, 1758).

The erythroid lineage was studied in the flounder-gloss (Platichthys flesus Linnaeus, 1758) during the annual cycle. The erythrocyte count in the blood was determined along with the contents of immature erythroid forms (basophilic and polychromatophilic normoblasts) in the head kidney (pronephros) and the blood. Cell proliferative activity was inferred from the [3H]thymidine inclusion in circulating immature erythrocytes. Irregularity was observed in erythropoiesis occurring in flounder-gloss hematopoietic tissue. Intense production of erythroid mass was mainly associated with a post-spawning period. This was evident from an increase in the contents of immature erythroid forms in the pronephros and circulating blood and an increase in their proliferative activity. The changes were associated with peculiarities of the erythroid system organization, which precludes regular production of erythropoietin in the kidney in teleost fish.

Comparison of [3H]-Thymidine, Carboxyfluorescein Diacetate Succinimidyl Ester and Ki-67 in Lymphocyte Proliferation.

Background: Patients with T cell deficiency <10% of normal proliferation are indicated to receive immune reconstruction by hematopoietic stem cell transplantation (HSCT). This study aimed to investigate whether non-radioactive assays can be used to quantitatively detect the lymphocyte proliferation <10% of normal as radioactive [3H]-thymidine." Methods: Radioactive [3H]-thymidine, non-radioactive carboxyfluorescein diacetate succinimidyl ester (CFSE), and Ki-67 protein expressions were used to measure the lymphocyte proliferation as calculated using the stimulation index (SI), subtraction percentage, and proliferation index (FlowJo software). Normal references were established for comparison in the absence of parallel healthy controls. Results: Normal ranges of mitogen-stimulated lymphocyte proliferation were established as a SI of 15-267 (CSFE 47-92%, Ki-67 42-79%) with phytohemagglutinin (PHA) 5 µg/ml stimulation; 19-139 (CFSE 62-83%, 45-74% Ki-67) with concanavalin-A (ConA) 5 µg/ml stimulation; 7-53 (CFSE 6-23%, Ki-67 10-24%) with pokeweed mitogen (PWM) 0.1 ug/ml stimulation; 3-28 (CFSE 4-10%, Ki-67 5-14%) with candida 10 ug/ml stimulation; and 2-27 (CFSE 6-41%, Ki-67 6-30%) with bacille Calmette-Guerin (BCG) 0.02 ng/ml stimulation. The normalized CFSE-proliferation index was between 2.1 and 3.0. Although there was no significant correlation between these three assays in the healthy controls, the SI value for <10% [3H]-thymidine proliferation in those with T cell deficiency was compatible with CFSE- and Ki-67-stained lymphocyte percentages, and validated in patients with IL2RG, RAG1, and ZAP70 mutations. When calculating [3H]-thymidine <10% of normal lymphocyte proliferation, the threshold of parallel controls was more reliable than previously established normal references. Conclusion: The large quantitative value of radioactive [3H]-thymidine was more easily recognizable than that for non-radioactive CFSE and Ki-67. Even though the correlation was not significant, those identified to have <10% of normal proliferation by [3H]-thymidine could be consistently detected by CFSE and Ki-67, and consequently indicated for HSCT.

Comparative Study of the γH2AX Foci Forming in Human Lung Fibroblasts Incubated in Media Containing Tritium-Labeled Thymidine or Amino Acids.

We compared the formation of γH2AX foci (marker of DNA double-strand breaks) in human lung fibroblasts (MRC-5 line) during their 24-h incubation in a medium containing 3H-labeled thymidine or amino acids (glycine, alanine, and proline) with specific radioactivity from 100 to 400 MBq/liter. A linear dependence of changes in the number of γH2AX foci on the specific radioactivity of the medium was revealed. The quantitative yield of DNA double-strand breaks under the influence of 3H-thymidine was more than 2-fold higher than under the influence of 3H-labeled amino acids. Comparative analysis of the yields of DNA double-strand breaks during cell incubation with 3H-labeled amino acids showed that 3H-alanine produced more pronounced effect that 3H-proline, which is consistent with the data on the content of their non-radioactive analogs in chromatin proteins.

Curcumin stimulates angiogenesis through VEGF and expression of HLA-G in first-trimester human placental trophoblasts.

Curcumin has a protective role in placental diseases like preeclampsia and preterm birth. Very little is known about its functional effects on growth, angiogenesis, and epigenetic activities of human first trimester placenta. HTR8/SVneo trophoblasts cells were used as model for human first trimester placenta. Effects of curcumin (≥80%) in these cells were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), radioactive thymidine uptake, quantitative real-time polymerase chain reaction (qRT-PCR), promoter DNA methylation, qRT-PCR array, tube formation, wound healing, and immunoblot assays. PC3 (prostate cancer), JEG-3 (trophoblast), and HMEC-1 (endothelial) cells were used as control in various experiments. Unlike in PC3 cells, curcumin stimulated growth, proliferation, and viability in HTR8/SVneo cells. Curcumin increased tube formation, and messenger RNA (mRNA) expression of angiogenic factors such as vascular endothelial growth factor A (VEGFA) and protein expression of proangiogenic factor VEGF receptor-2 and fatty acid-binding protein-4 (FABP4) in these cells. Curcumin-stimulated tube formation was associated with an increased expression of VEGFR2 and FABP4. The stimulatory effects of curcumin were inhibited by VEGFR2 (SU5416) and FABP4 (BMS309403) inhibitors. Curcumin also significantly increased both mRNA and protein expression of HLA-G in HTR8/SVneo cells. Curcumin increased mRNA expression of DNMT3A and NOTCH signaling system whereas down-regulated mRNA expression of HSD11ß2. Curcumin enhanced hypomethylation of gene promoters against oxidative stress and DNA damage pathway mediators. Curcumin promotes cell growth, migration, and thus angiogenic potential of these cells. Increased expression of HLA-G by curcumin, hitherto unknown, is a novel finding since HLA-G not only favors the immune environment for invasive trophoblasts but also positively modulates angiogenesis.

Comparative Analysis of the Formation of γH2AX Foci in Human Mesenchymal Stem Cells Exposed to 3H-Thymidine, Tritium Oxide, and X-Rays Irradiation.

We performed a comparative study of the formation of γН2АХ foci (a marker of DNA doublestrand breaks) in human bone marrow mesenchymal stem cells after 24-h incubation with 3Н-thimidin and tritium oxide with low specific activities (50-800 MBq/liter). The dependence of the number of γH2AX foci on specific activity of 3H-thymidine was described by a linear equation y=2.21+43.45x (R2=0.96), where y is the number of γH2AX foci per nucleus and x is specific activity in 1000 MBq/liter. For tritium oxide, the relationship was described by a linear equation y=2.52+6.70x (R2=0.97). Thus, the yield of DNA double-strand breaks after exposure to 3H-thymidine was 6.5-fold higher than after exposure to tritium oxide. Comparison of the effects of tritium oxide and X-ray radiation on the yield of DNA double-strand breaks showed that the relative biological efficiency of tritium oxide in a dose range of 3.78-60.26 mGy was 1.6-fold higher than that of X-ray radiation. Improvement of the methods of analysis of DNA double-strand breaks repair foci is highly promising in the context of creation of highly sensitive biodosimetry technologies for tritium compounds in humans.

Clinical Correlation of the Histoculture Drug Response Assay for Head and Neck Cancer.

Gelfoam® histoculture was utilized to develop the histoculture drug response assay (HDRA) for head and neck cancer. Specimens of head and neck tumors were evaluated for sensitivity to the following drugs: cisplatinum (CDDP), 5-fluorouracil (5-FU), and the combination of CDDP and 5-FU. In the first clinical study at UCSD, 10 of 12 patients with tumors that were drug sensitive in Gelfoam® histoculture had either complete or partial response clinically. Comparisons of HDRA results, obtained with [3H]thymidine incorporation as the endpoint were made with clinical responses, i.e., complete response, partial response, or no response. The overall accuracy of the HDRA was 74% in this correlative clinical trial; the predictive positive value was 83%, the sensitivity was 71%, and the specificity was 78%. Seven of 11 patients with HDRA-resistant tumors demonstrated no response for a predictive negative value of 64%. In a subsequent study at Memorial Sloan Kettering Cancer Center, tumor specimens from 41 to 42 patients undergoing treatment for head and neck cancer were successfully evaluated by the HDRA. The histocultured tumors were treated with 5-FU and/or CDDP and a control group received no drug treatment. After completion of drug treatment, the relative cell survival in the tumors was determined using the MTT endpoint. Sensitivity was defined as a tumor inhibition rate (IR) of greater than 30%. Survival comparisons were performed using the generalized Wilcoxon test for the comparison of Kaplan-Meier survival curves. Resistance to 5-FU was observed in 13 cases (32%), to CDDP in 13 cases (32%), and to both agents in 11 cases (27%). The 2-year cause-specific survival was significantly greater for patients sensitive to 5-FU than patients who were resistant (85% vs. 64%), CDDP (86% vs. 64%), or both agents (85% vs. 63%). These results demonstrate the clinical usefulness of the HDRA for head and neck cancer.

Dual Antibacterial and Anticancer Activity of 4-Benzoyl-1-dichlorobenzoylthiosemicarbazide Derivatives.

OBJECTIVE/METHOD: A group of 4-benzoyl-1-dichlorobenzoylthiosemicarbazides endowed with antibacterial activity was evaluated for its cytotoxic properties against breast cancer cells (MCF-7, MDA-MB-231) and head and neck squamous cell carcinomas (FaDu, SCC-25). Cytotoxicity of the investigated compounds was measured using MTT and [3H]-thymidine incorporation bioassays. RESULT: 1-(2,3-Dichlorobenzoyl)-4-(2-methylbenzoyl)thiosemicarbazide (TA-4), 1-(2,4-dichlorobenzoyl)- 4-(2-methylbenzoyl)thiosemicarbazide (TA-18), and 1-(2,4-dichlorobenzoyl)-4-(4-nitrolbenzoyl)- thiosemicarbazide (TA-20) were found to possess anticancer activity equipotent or even stronger than that of reference drug - etoposide. In order to clarify the molecular mode of action of the mentioned compounds, the relaxation assay kit for human DNA topoisomerase II was used. It turned out that reduction of viability of cancer cells was a result of inhibition of human DNA topoII. Molecular docking studies proved that 4-benzoyl-1-dichlorobenzoylthiosemicarbazides strongly interact with DNAdependent subunit of that enzyme.

In vitro studies on the inhibition of colon cancer by amino acid derivatives of bromothiazole.

The investment in cancer research is critical to find more and better treatments, but essentially to save lives. Here, we describe the synthesis and characterization on new bromothiazole derivatives with amino acids and with core of nitazoxanide, an FDA-approved antiprotozoal drug. Using a human adenocarcinoma-derived cell line (the Caco-2 cell line), we then investigated the antiproliferative (3H-thymidine incorporation) and cytotoxic (extracellular lactate dehydrogenase activity) effect of these derivatives. All the derivatives caused a concentration-dependent decrease in cell proliferation and viability. At their highest concentration, all compounds were able to reduce 3H-thymidine incorporation by more than 80%, corresponding to a more marked antiproliferative effect than butyrate. As to their cytotoxic effect, it was comparable to that of butyrate. The ability of bromo substituent in thiazole ring with new sequences of amino acids in inducing cell death and apoptosis in Caco-2 cells (and other cell lines) is now being studied.

Assessment of lymphocyte proliferation for diagnostic purpose: Comparison of CFSE staining, Ki-67 expression and 3H-thymidine incorporation.

The capability of lymphocytes to respond to antigenic or mitogenic stimulation is an important feature in the diagnosis of various immunodeficiencies and immune disorders. We used large cohorts of both immune compromised patients and healthy controls to measure lymphocyte proliferations by means of three methods: CFSE staining, Ki-67 expression and 3H-thymidine incorporation. The advantages and disadvantages of each method was then evaluated for use in routine clinical diagnostic. The statistical analysis was performed between the outcomes and the correlation between all three methods was computed. CFSE and Ki-67 assay correlated well with the r=0.767, correlation between Ki-67 expression and 3H-thymidine incorporation was 0.546 and correlation between CFSE staining and 3H-thymidine incorporation was 0.337. The differences between these three methods concerning complexity, sensitivity and reliability as well as the financial aspects are discussed hereafter. CFSE and its analogues provide the cheapest and reasonable choice for measuring lymphocyte proliferation, while Ki-67 represents a more expensive, but more sensitive and robust method. The original 3H-thymidine assay does not bring any advantages and cannot compare to the competition presented by modern flow cytometric methods available today.

Effects of Malignant Melanoma Initiating Cells on T-Cell Activation.

Although human malignant melanoma is a highly immunogenic cancer, both the endogenous antitumor immune response and melanoma immunotherapy often fail to control neoplastic progression. Accordingly, characterizing melanoma cell subsets capable of evading antitumor immunity could unravel optimized treatment strategies that might reduce morbidity and mortality from melanoma. By virtue of their preferential capacity to modulate antitumor immune responses and drive inexorable tumor growth and progression, malignant melanoma-initiating cells (MMICs) warrant closer investigation to further elucidate the cellular and molecular mechanisms underlying melanoma immune evasion and immunotherapy resistance. Here we describe methodologies that enable the characterization of immunoregulatory effects of purified MMICs versus melanoma bulk populations in coculture with syngeneic or allogeneic lymphocytes, using [3H]thymidine incorporation, enzyme-linked immunosorbent spot (ELISPOT), or ELISA assays. These assays were traditionally developed to analyze alloimmune processes and we successfully adapted them for the study of tumor-mediated immunomodulatory functions.

Generation and vulnerability of deep cerebellar nuclei neurons in the weaver condition along the anteroposterior and mediolateral axes.

Production and death of deep cerebellar nuclei (DCN) neurons were investigated in the weaver condition at appropriate anatomical levels throughout the mediolateral (medial, intermediate and lateral) and rostrocaudal (rostral, middle and caudal) axes of three DCN-cell groups: the fastigial, the interposed and the dentate nuclei. Current results have denoted that the deficit of DCN neurons is always more important in the homozygous weaver than in the heterozygous weaver mice. No loss of neurons was found in the dentate nucleus. In the mediolateral axis, an intranuclear gradient of depletion was observed in the mutant mice; in a given deep nucleus, neurodegeneration was more prominent in the medial pars than in lateral ones. In the rostrocaudal axis, on the other hand, when each deep nucleus was studied and compared as a whole, neuron loss was higher in the fastigial nucleus than in the interposed nucleus, which, in turn, was more important than in the dentate nucleus. These data suggest that, in the weaver condition, an internuclear gradient of neurodegeneration exists. Moreover, neurons located in rostral parts of a given nucleus appear to be more vulnerable than those settled in middle parts and these, in turn, are more than the caudal ones. These results seem to indicate the presence of an intranuclear gradient of depletion. Current autoradiographic results have revealed that, in the rostrocaudal axis, deep neurons are settled in the weaver cerebellum following three neurogenetic gradients. The first of these is internuclear; if each deep nucleus is analyzed and compared as a whole, the fastigial nucleus has more late-generated neurons than the interposed nucleus, and this, in turn, has more than the dentate nucleus. The second gradient is also internuclear; if the proportion of late-born neurons is compared throughout the rostral levels from each deep nucleus, it is observed that proportions increase from the fastigial to the dentate nucleus. A similar picture emerges when the middle and caudal regions are taken into account. The third gradient is intranuclear; in a given deep nucleus, the rostral region always presents more late-produced neurons than the middle region and these, in turn, more than in the caudal level.

Systematic differences in time of cerebellar-neuron origin derived from bromodeoxyuridine immunoperoxidase staining protocols and tritiated thymidine autoradiography: A comparative study.

As exogenous markers of DNA synthesis, 5-bromo-2'-deoxyuridine (BrdU) and tritiated thymidine ([(3)H]TdR) have revolutionized our ability to identify proliferating neuroblasts and follow their fate during the development of the central nervous system. The effect of the incorporation of these molecules into DNA on cell proliferation, migration and differentiation is frequently neglected (Duque and Rakic, 2011. J. Neurosci. 31, 15205-15217). By a progressively delayed cumulative labeling method, the current paper analyzes the development of the cerebellum in mice exposed to either BrdU or [(3)H]TdR as embryos and collected at postnatal day 90. We observed that, in comparison to the saline group, several parameters of the cerebellum such as length of the cerebellar cortex, the area of the molecular layer, Purkinje cell (PCs) number, the areas of the cerebellar nuclei, and the number of the deep cerebellar nuclei (DCN) neurons were lower in the BrdU injected group. No consequence of [(3)H]TdR administration was observed. On the other hand, we also studied whether immunohistochemical methods, including BrdU antibodies from different vendors (Sigma and Dako), partial DNA denaturation procedures and trypsin pretreatments, alter the neurogenetic timetables of PC and DCN neurons that resulted from analysis of these tissue specimens. Our analysis revealed that the generative programs of these macroneurons were unrelated to differences in the sensibility of BrdU antibodies but were dependent on the partial denaturation of DNA and trypsin digestion protocols. Finally, we also compare the generation and spatial distribution of PC and DCN neurons in mice exposed to either BrdU or [(3)H]TdR to assess whether the results obtained by these two markers are quantitatively similar. The data presented here show that systematic differences exist in the pattern of neurogenesis and the spatial location of cerebellar neurons between mice injected with BrdU or [(3)H]TdR. These findings have implications for the interpretation of results obtained by both exogenous makers as an index of the production, migration and settling of neurons in the developing central nervous system.

Flow cytometric evaluation of lymphocyte transformation test based on 5-ethynyl-2'deoxyuridine incorporation as a clinical alternative to tritiated thymidine uptake measurement.

In clinical laboratories, the evaluation of lymphocyte proliferative response (lymphocyte transformation test-LTT) is routinely performed by the measurement of [(3)H]-thymidine uptake after stimulation. In this study we evaluated the performances of a recently developed non-radioactive test based on the detection by flow cytometry of 5-ethynyl-2'deoxyuridine (EdU) incorporation for the measurement of LTT in routine lab conditions. After definition of optimal protocol parameters, EdU incorporation test showed good repeatability and reproducibility. Moreover, this assay was flexible enough to fit important clinical laboratory constraints (delayed stimulation, low number of cells and delayed analysis after staining). Importantly, correlations between results obtained with EdU and [(3)H]-thymidine incorporation assays were excellent both in healthy volunteers and pediatric and septic patients. In particular, the two techniques identified patients presenting with altered LTT. Upon confirmation in a larger cohort of patients, EdU incorporation assay may be a relevant non-radioactive candidate for LLT in clinic.

Administering the Optimum Dose of l-Arginine in Regional Tumor Therapy.

The purpose of this study is optimizing the l-arginine (l-Arg) doses on the basis of chemical structure in regional accessible tumor therapy to settle down a new protocol for the treatment of cancer. (3)H-thymidine-based cell proliferation assay was performed in vitro on tumor cell lines of fibrosarcoma (FS), lymphosarcoma-ascitic and on normal cell line of NIH 3T3 after treatment with different concentrations of l-Arg in phosphate buffered saline (PBS). The cultures were harvested after 22 h and the incorporated radioactivity was counted to identify their histologic grades as described in earlier studies. In vivo therapy of murine tumors was conducted where FS cells injected subcutaneously at ventro-lateral position of mice. Various drug delivery schedules were injected into the centre of tumor base, once a day for 4 days. Tumor diameter and survivals were monitored where the day of sacrifice was considered for monitoring the survival period. By identifying the histologic grades of the treated cultures in vitro and in vivo by different concentrations of l-Arg, the corresponding energy of such concentrations were determined. An efficient model with a good fit (R(2) = 0.98) was established to describe the energy yield by l-Arg dose. The equivalence between the tumor histologic grade and energy of the l-Arg dose delivered in saline (PBS) environment is the optimum condition for regional tumor therapy achieves higher survival rate. The selective cytotoxicity to tumor cells with minimal damage to normal cells by l-Arg due to its chemical structure suggests to be considered the most promising drug for regional therapy of the accessible tumors like breast cancers of early stage with no distant metastasis.

The effects of arsenic trioxide on DNA synthesis and genotoxicity in human colon cancer cells.

Colon cancer is the third leading cause of cancer-related deaths worldwide. Recent studies in our laboratory have demonstrated that arsenic trioxide is cytotoxic in human colon cancer (HT-29), lung (A549) and breast (MCF-7) carcinoma cells. The purpose of the present study is to investigate the effects of arsenic trioxide on DNA synthesis and the possible genotoxic effects on human colon cancer cells. HT-29 cells were cultured according to standard protocol, followed by exposure to various doses (0, 2, 4, 6, 8, 10, and 12 microg/mL) of arsenic trioxide for 24 h. The proliferative response (DNA synthesis) to arsenic trioxide was assessed by [(3)H]thymidine incorporation. The genotoxic effects of arsenic-induced DNA damage in a human colon cancer cell line was evaluated by the alkaline single cell gel electrophoresis. Results indicated that arsenic trioxide affected DNA synthesis in HT-29 cells in a biphasic manner; showing a slight but not significant increase in cell proliferation at lower levels of exposure (2, 4 and 6 microg/mL) followed by a significant inhibition of cell proliferation at higher doses (i.e., 8 and 10 microg/mL). The study also confirmed that arsenic trioxide exposure caused genotoxicity as revealed by the significant increase in DNA damage, comet tail-lengths, and tail moment when compared to non-exposed cells. Results of the [(3)H]thymidine incorporation assay and comet assay revealed that exposure to arsenic trioxide affected DNA synthesis and exhibited genotoxic effects in human colon cancer cells.

Effects of Adriamycin or CP -2 on the Spleen of Mouse Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 체질인류학회지

In this experiment, side effects of two anticancer drugs (adriamycin and CP -2) on the structure of spleen were histologically studied. Each of ICR mice was inoculated with 1 x10 7 Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline solution, adriamycin (2 mg/kg) or CP -2 (30 mg/kg) were injected subcutaneously every other day. The day following the 7th injection of adriamycin or CP -2, each mouse was injected with a single dose of 0.7 micro Ci/gm of methyl -3 H -thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and splenic tissues were collected and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM -1 (Amersham Lab., England) in the dark room and dried, and were kept in a light -tight box. The sections were exposured for 5 weeks in the dark room, and were developed in D -19 developer. The number of the labeled cells in the areas of the white pulp, the red pulp and the marginal zone (mean number of labeled cells per 0.21 mm 2 ) were observed and calculated. In the spleen of adriamycin treated group, vacuoles containing pyknotic nuclei were observed frequently. Whereas in the CP -2 treated group, morphological changes of the spleen were not observed. The number of the labeled cells of normal control, experimental control, CP -2 treated and adriamycin treated groups were 240.3 +/-53.28, 252.3+/- 58.24, 216.7 +/-55.17 and 45.4 +/-15.46, respectively, and most of the labeled cells were located near the marginal zone of the spleen. In the adriamycin treated group, labeled cells containing a few silver grains of 3 H -thymidine were observed more frequently than in those of the normal and experimental control groups. From the above results, adriamycin and CP -2 may suppress the DNA synthesis of the splenic tissues. Especially, CP -2 does not results any histological defect on the splenic tissues. These result suggest that CP -2 is expected as one of effective anticancer drugs.

The Effect of Antimetabolites for Inhibiting the Proliferation of Rabbit Lens Epithelial Cells in Vitro

The most common cause of blurred vision after extracapsular cataract extraction is known to be an opacification of the posterior lens capsule. The pathogenesis of posterior lens capsule opacification is primarily caused by residual lens epithelial cells. For the prevention of posterior capsular opacification, several kinds of anti-mitotic drugs is being actively investigated. But the antimitotic drugs are not clinically used due to toxicity towards the intraocular tissues. The objectives of this study is to evaluate the effect of mitomycin C and tirilazad mesylate(FREEDOX(TM)) respectively for inhibiting the proliferation of rabbit lens epithelial cells when it is administered in a short period. Lens epithelial cells from white rabbits were harvested andcultured for 4 passages. Mitomycin C was applied for 3 minutes with 0.025mg/ml and 0.05mg/ml in concentration respectively. The proliferation assay was performed by [(3)H]-thymidine uptake test. Significant decrease of lens epithelial cell proliferation appeared in both drugs.When Mitomycin-C was applied with 0.025mg/ml for 3 minutes, cell proliferation was reduced to 31.5% compared with control and in 0.05mg/ml concentration, to 12.5%. When tirilazad mesylate was applied 0.15mg/ml for 3 minutes, cell proliferation was reduced to 46.5% compared with control and in 1.5mg/ml concentration, to 7.5%. If futher investigation would show the effectives and safety of these drugs, these agents could be applied into the lens capsular bad at the time of surgery to prevent the posterior capsular opacification after cataract surgery.

A histologic and autoradiographic study on the early changes in the movements of rat incisors / 대한치과교정학회지

The purpose of this study was to quantify the biologic effects of the tensile forces from helical springs across the maxillary incisors on the periodontal tissues of rats. 39 Sprague-Dawlely rats were divided into a control group(3 rats) and three experimental groups(36 rats)-group 1, pressured with a light force(50-75g), group 2, with a heavy force(250-300g) and group 3, with a heavy force(250-300g) plus laser irradiation. Autoradiographic and histopathologic observations were performed in 12, 24, 48 and 96 hours after force application. The result were as follow: 1. Hyalinized zone of periodontal ligament began to appear at pressure side in 12 hours in group 2 and group 3 ; all decreased in 96 hours except in group 2. 2. Alveolar bone resorption began to appear in 12 hours in group 2 and group 3 ; Group 2 showed more resorption than group 1 and no difference to group 3. 3. Tearing of periodontal ligament and vascular dilatation began to appear at tension side in 12 hours in all groups ; Group 2 showed more changes than group 1 and no difference to group 3 ; Decrease began to appear in 96 hours. 4. New bone formation began to appear at tension side in 12 hours and increased more and more; No differences were shown of groups 5. New capillary proliferation began to appear at pressure side in 12 hours; The changes were the greatest in group 3, group 2, group 1, in that order. 6. Positive reaction of cells to [3H]-thymidine was the greatest in 24 hours of all groups and decreased with times ; Group 2 showed more reaction than group 1 and no difference to group 3.

Effects of IL-7 on human lymphocytes

Interleukin-7 (IL-7) is known as a growth factor for pre B-cell and mature T-cells in human. But in leukemic cells, IL-7 effect is variously reported. To investigate the effect of IL-7 on the cells of childhood acute leukemia we used 3H-Thymidine assay. Twelve Acute lymphoblastic leukemia (ALL), seven T-ALL and three Acute myelogenous leukemia (AML) were involved in this study. Two out of twelve ALL and three out of seven T-ALL bone marrow (BM) cells were stimulated by IL-7 in 3H-Thymidine incorporation. In normal and AML BM cells, IL-7 had no stimulatory activity as in various leukemic cell lines. Two normal peripheral blood T-cells responded to IL-7 dose dependently. We have seen the effect of IL-7 to stimulate T-lineage cells but, for precise conclusion, further study using more purified samples will be needed.

EFFECTS OF VERAPAMIL ON DNA, RNA AND PROTEIN SYNTHESIS OF HUMAN FIBROBLASTS / 中国药理学通报

In order to investigate the action of calcium antagonists on metabolism of intracellular macromolecules, The authors observed the effects of verapamil on the incor-poration of 〔 3H 〕 TdR, 〔 3H 〕 UR and 〔 3H 〕 Leu into human fibroblasts. The inhibitory effects on DNA and RNA synthesis were concentration dependent, ID50 was l2.3mg/L and 22mg/L, respectively. The inhibitory effect on the synthesis of protein was weak.