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Rationale: Fetal growth and survival depend critically on proper development and integrity of the vascular system. Fli1 (Friend leukemia integration 1), a member of the Ets family of transcription factors, plays critical roles in vascular morphogenesis and homeostasis at mid-gestation, the developmental stage at which expression of its upstream regulator, Etv2, ceases. However, molecular mechanisms of Fli1 action in vascular morphogenesis remain incompletely understood. Objective: To dissect molecular mechanisms of vascular morphogenesis governed by Fli1. Methods and Results: Utilizing Fli1 promoter-driven lineage-specific LacZ expression, Fli1 loss-of-function strategies, and a series of molecular techniques, we demonstrate that Fli1 expression in multipotent myogenic progenitor cells (MPCs) occurs independent of Etv2, and loss of Fli1 expression results in a significant increase in LacZ + cells in mesoderm within somites and limb buds, leading to reciprocal regulation of the expression of several key endothelial and myogenic genes and vascular abnormalities. Conversely, embryos harboring conditional Fli1 gain-of-function in MPCs manifested aberrant vasculogenesis with lack of myogenesis. Mechanistically, elevated Fli1 activity in myoblasts and in adult MPCs (also called satellite cells) of X-linked muscular dystrophic mdx mice markedly induced endothelial, but attenuated myogenic, gene expression and differentiation. Importantly, ectopic expression of Myf5 or MyoD, two key myogenic regulators, in Fli1-expressing myoblasts restored their differentiation potential, indicating that levels of Fli1 and myogenic regulators in MPCs inversely regulate their endothelial versus myogenic potential. Conclusions: Fli1 governs vascular morphogenesis by regulating endothelial potential by inversely regulating endothelial versus myogenic programs in MPCs. Our data uncover an important and previously unrecognized mechanism of vascular morphogenesis governed by Fli1 and highlight the physiological significance of the fine tuning of Fli1 activity in multipotent progenitors for proper vascular and muscle morphogenesis during development and disease.

Particle-induced X-ray emission (PIXE) method was used in this present research to identify the elements present in selected vegetable samples to show the possible influence in the metal absorption by the vegetables grown in a saline region of Rampal area of Munshiganj District, Bangladesh. The data acquisition setup is calibrated using a 2.5-MeV proton beam in the current ranges of 5nA to 15nA. The detector was used to measure the X-rays emitted during the irradiation. Data acquisition system MAESTRO-32 was used to measure the spectrum picks, and concentration calculation has been done by GUPIX/DAN-32 software. The purpose of this study is to determine the concentration of heavy and trace elements in these samples and to give current information on their safety for consumption. The result shows that K, Ca, and Fe have the highest concentrations, while lead exhibits the lowest but alarming rates compared to reference materials. The findings were likened to IAEA-V-10, IAEA-359, SRM 1515 (apple leaf), and SRM 1573a (tomato leaf). The outcomes of the present investigation demonstrate that these samples are not devoid of health risks in intake.

CK2 is a master regulator protein kinase which demonstrates heightened expression in diverse cancer types and is considered a promising target for therapy. Given its ubiquitous expression and potent influence on cell survival, cancer cell-directed targeting of the CK2 signal is an important factor for development of an anti-CK2 therapeutic. We previously reported on the malignant cell specificity and effect on CK2 signaling of a tenfibgen (TBG) based nanocapsule for delivery of the CK2 small molecule inhibitor 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT) in cultured prostate cancer cells. Here we tested the ability of TBG-DMAT to affect the growth of prostate xenograft tumors in mice. Our results show that treatment of PC3-LN4 xenograft tumors with TBG-DMAT caused loss of proliferative Ki-67 signal as well as Nuclear Factor-kappa B (NF-κB) expression in the tumors. Further, the TBG-DMAT nanocapsule was detected in tumors and not in liver or testis. In conclusion, TBG-based nanocapsule delivery of anti-CK2 small molecule drugs holds significant promise for treatment of prostate cancer.

Multiword detection is very difficult task in Language Processing. There has been a great change in the field of Natural Language Processing. Manual encoding of linguistic information is being challenged by automated corpus based learning methodologies for NLP with linguistic Knowledge. Although, Corpus based approaches have been successful in many different areas of Natural Language Processing. Multiword Detection using human evaluation method and machine evaluation method is a matter of discussion in present NLP research areas. Languages are not use not only to gather information but for communication as well. The problem is to take the information provided by the outside the world and translate the information into precise internal representation .This internal representation is called semantic representation. In this paper, we deals with how MWEs are generalized to optimized expensive evaluation of Multiword detection from a WordNet.

Deceleration in rice ( Oryza sativa L.) yield over time under fixed management conditions is a concern for countries like Bangladesh, where rice is the primary source of calories for the human population. Field experiments were conducted from 1990 to 1999 on a Chhiata clay loam soil (Hyperthermic Vertic Endoaquept) in Bangladesh, to determine the effect of different doses of chemical fertilizers alone or in combination with cow dung (CD) and rice husk ash (ash) on yield of lowland rice. Two rice crops—dry season rice (December–May) and wet season rice (July–November) were grown in each year. Six treatments—absolute control (T 1 ), one-third of recommended fertilizer doses (T 2 ), two-thirds of recommended fertilizer doses (T 3 ), full doses of recommended fertilizers (T 4 ), T 2 +5 t CD and 2.5 t ash ha −1 (T 5 ) and T 3 +5 t CD and 2.5 t ash ha −1 (T 6 ) were compared. The CD and ash were applied on dry season rice only. The 10-year mean grain yield of rice with T 1 was 5.33 t ha −1 per year, while the yield with T 2 was 6.86 t ha −1 per year. Increased fertilizer doses with T 3 increased the grain yield to 8.07 t ha −1 per year, while the application of recommended chemical fertilizer doses (T 4 ) gave 8.87 t ha −1 per year. The application of CD and ash (T 5 and T 6 ) increased rice yield by about 1 t ha −1 per year over that obtained with chemical fertilizer alone (T 2 and T 3 , respectively). Over 10 years, the grain yield trend with the control plots was negative, but not significantly, both in the dry and wet seasons. Under T 3 through T 6 , the yield trend was significantly positive in the dry season, but no significant trend was observed in the wet season. The treatments, which showed positive yield trend, also showed positive total P uptake trend. Positive yield trends were attributed to the increasing P supplying power of the soil.

Galectin-9 (Gal-9) induced the apoptosis of not only T cell lines but also of other types of cell lines in a dose- and time-dependent manner. The apoptosis was suppressed by lactose, but not by sucrose, indicating that β-galactoside binding is essential for Gal-9-induced apoptosis. Moreover, Gal-9 required at least 60 min of Gal-9 binding and possibly de novo protein synthesis to mediate the apoptosis. We also assessed the apoptosis of peripheral blood T cells by Gal-9. Apoptosis was induced in both activated CD4+ and CD8+ T cells, but the former were more susceptible than the latter. A pan-caspase inhibitor (Z-VAD-FMK) inhibited Gal-9-induced apoptosis. Furthermore, a caspase-1 inhibitor (Z-YVAD-FMK), but not others such as Z-IETD-FMK (caspase-8 inhibitor), Z-LEHD-FMK (caspase-9 inhibitor), and Z-AEVD-FMK (caspase-10 inhibitor), inhibited Gal-9-induced apoptosis. We also found that a calpain inhibitor (Z-LLY-FMK) suppresses Gal-9-induced apoptosis, that Gal-9 induces calcium (Ca2+) influx, and that either the intracellular Ca2+ chelator BAPTA-AM or an inositol trisphosphate inhibitor 2-aminoethoxydiphenyl borate inhibits Gal-9-induced apoptosis. These results suggest that Gal-9 induces apoptosis via the Ca2+-calpain-caspase-1 pathway, and that Gal-9 plays a role in immunomodulation of T cell-mediated immune responses.

Introduction Targeted therapies for aggressive breast cancers like triple negative breast cancer (TNBC) are needed. The use of small interfering RNAs (siRNAs) to disable expression of survival genes provides a tool for killing these cancer cells. Cyclin dependent kinase 11 (CDK11) is a survival protein kinase that regulates RNA transcription, splicing and mitosis. Casein kinase 2 (CK2) is a survival protein kinase that suppresses cancer cell death. Eliminating the expression of these genes has potential therapeutic utility for breast cancer. Methods Expression levels of CDK11 and CK2 mRNAs and associated proteins were examined in breast cancer cell lines and tissue arrays. RNA expression levels of CDC2L1, CDC2L2, CCNL1, CCNL2, CSNK2A1, CSNK2A2, and CSNK2B genes in breast cancer subtypes were analyzed. Effects following transfection of siRNAs against CDK11 and CK2 in cultured cells were examined by viability and clonal survival assays and by RNA and protein measures. Uptake of tenfibgen (TBG) nanocapsules by TNBC cells was analyzed by fluorescence-activated cell sorting. TBG nanocapsules delivered siRNAs targeting CDK11 or CK2 in mice carrying TNBC xenograft tumors. Transcript cleavage and response parameters were evaluated. Results We found strong CDK11 and CK2 mRNA and protein expression in most human breast cancer cells. Immunohistochemical analysis of TNBC patient tissues showed 100% of tumors stained positive for CDK11 with high nuclear intensity compared to normal tissue. The Cancer Genome Atlas analysis comparing basal to other breast cancer subtypes and to normal breast revealed statistically significant differences. Down-regulation of CDK11 and/or CK2 in breast cancer cells caused significant loss of cell viability and clonal survival, reduced relevant mRNA and protein expression, and induced cell death changes. TBG nanocapsules were taken up by TNBC cells both in culture and in xenograft tumors. Treatment with TBG- siRNA to CDK11 or TBG- siRNA to CK2αα’ nanocapsules induced appropriate cleavage of CDK11 and CK2α transcripts in TNBC tumors, and caused MDA-MB-231 tumor reduction, loss of proliferation, and decreased expression of targeted genes. Conclusions CDK11 and CK2 expression are individually essential for breast cancer cell survival, including TNBC. These genes serve as promising new targets for therapeutic development in breast cancer.

The use of arsenic (As) contaminated groundwater for irrigation of crops has resulted in elevated concentrations of arsenic in agricultural soils in Bangladesh, West Bengal (India), and elsewhere. Paddy rice (Oryza sativa L.) is the main agricultural crop grown in the arsenic-affected areas of Bangladesh. There is, therefore, concern regarding accumulation of arsenic in rice grown those soils. A greenhouse study was conducted to examine the effects of arsenic-contaminated irrigation water on the growth of rice and uptake and speciation of arsenic. Treatments of the greenhouse experiment consisted of two phosphate doses and seven different arsenate concentrations ranging from 0 to 8 mg of As L(-1) applied regularly throughout the 170-day post-transplantation growing period until plants were ready for harvesting. Increasing the concentration of arsenate in irrigation water significantly decreased plant height, grain yield, the number of filled grains, grain weight, and root biomass, while the arsenic concentrations in root, straw, and rice husk increased significantly. Concentrations of arsenic in rice grain did not exceed the food hygiene concentration limit (1.0 mg of As kg(-1) dry weight). The concentrations of arsenic in rice straw (up to 91.8 mg kg(-1) for the highest As treatment) were of the same order of magnitude as root arsenic concentrations (up to 107.5 mg kg(-1)), suggesting that arsenic can be readily translocated to the shoot. While not covered by food hygiene regulations, rice straw is used as cattle feed in many countries including Bangladesh. The high arsenic concentrations may have the potential for adverse health effects on the cattle and an increase of arsenic exposure in humans via the plant-animal-human pathway. Arsenic concentrations in rice plant parts except husk were not affected by application of phosphate. As the concentration of arsenic in the rice grain was low, arsenic speciation was performed only on rice straw to predict the risk associated with feeding contaminated straw to the cattle. Speciation of arsenic in tissues (using HPLC-ICP-MS) revealed that the predominant species present in straw was arsenate followed by arsenite and dimethylarsinic acid (DMAA). As DMAA is only present at low concentrations, it is unlikely this will greatly alter the toxicity of arsenic present in rice.

Elevated soil arsenic levels resulting from long-term use of arsenic contaminated ground for irrigation in Bangladesh may inhibit seed germination and seedling establishment of rice, the country's main food crop. A germination study on rice seeds and a short-term toxicity experiment with different concentrations of arsenite and arsenate on rice seedlings were conducted. Percent germination over control decreased significantly with increasing concentrations of arsenite and arsenate. Arsenite was found to be more toxic than arsenate for rice seed germination. There were varietal differences among the test varieties in response to arsenite and arsenate exposure. The performance of the dry season variety Purbachi was the best among the varieties. Germination of Purbachi was not inhibited at all up to 4 mg l−1 arsenite and 8 mg l−1 arsenate treatment. Root tolerance index (RTI) and relative shoot height (RSH) for rice seedlings decreased with increasing concentrations of arsenite and arsenate. Reduction of RTI caused by arsenate was higher than that of arsenite. In general, dry season varieties have more tolerance to arsenite or arsenate than the wet season varieties.

Long-term use of arsenic contaminated groundwater to irrigate crops, especially paddy rice (Oryza sativaL.) has resulted in elevated soil arsenic levels in Bangladesh. There is, therefore, concern regarding accumulation of arsenic in rice grown on these soils. A greenhouse pot experiment was conducted to evaluate the impact of arsenic-contaminated irrigation water on the growth and uptake of arsenic into rice grain, husk, straw and root. There were altogether 10 treatments which were a combination of five arsenate irrigation water concentrations (0–8 mg As l−1) and two soil phosphate amendments. Use of arsenate containing irrigation water reduced plant height, decreased rice yield and affected development of root growth. Arsenic concentrations in all plant parts increased with increasing arsenate concentration in irrigation water. However, arsenic concentration in rice grain did not exceed the maximum permissible limit of 1.0 mg As kg−1. Arsenic accumulation in rice straw at very high levels indicates that feeding cattle with such contaminated straw could be a direct threat for their health and also, indirectly, to human health via presumably contaminated bovine meat and milk. Phosphate application neither showed any significant difference in plant growth and development, nor in As concentrations in plant parts.

The experiment was conducted to measure crop yields, nutrient concentration, nutrient uptake and balance by using different nutrient management practices for mustard-mungbean-T. aman rice cropping system in calcareous soil of Madaripur, Bangladesh. Different nutrient management practices were absolute nutrient control (T1); farmer’s practice (T2); AEZ based nutrient application (T3) and soil test based nutrient application (T4). The practices were compared in a randomized completely block design with three replications over two consecutive years. The average yield through application of soil test based nutrient (T4) was showed effective to get highest yields of mustard (1530 kg ha-1), mungbean (1632 kg ha-1) and T. aman rice (4729 kg ha-1). The same practices (T4) exhibited the greatest nutrients uptake by the test crops. The apparent balance of N and K was negative; however it was less negative and less deficiency detect in T4 treatment. Positive balance of P observed in all practices except in T1. There was a positive S balance (7.60 kg ha-1) in T4 but negative in T1, T2 and T3. Zinc balance was found positive in T3 and T4 and negative in T1 and T2. Boron balance in the system was neutral or slightly positive in T1 and negative in T2 but positive in T3 and T4. Organic matter, N, P, S, Zn and B status in soil was improved by T4 treatment. The results suggested that the soil test based nutrient application is viable and sustainable for mustard-mungbean-T. aman rice cropping system in calcareous soils of Bangladesh.

Rationale: Cardiovascular health depends on proper development and integrity of blood vessels. Ets variant 2 (Etv2), a member of the E26 transforming–specific family of transcription factors, is essential to initiate a transcriptional program leading to vascular morphogenesis in early mouse embryos. However, endothelial expression of the Etv2 gene ceases at midgestation; therefore, vascular development past this stage must continue independent of Etv2. Objective: To identify molecular mechanisms underlying transcriptional regulation of vascular morphogenesis and homeostasis in the absence of Etv2. Methods and Results: Using loss- and gain-of-function strategies and a series of molecular techniques, we identify Friend leukemia integration 1 ( Fli1 ), another E26 transforming–specific family transcription factor, as a downstream target of Etv2. We demonstrate that Etv2 binds to conserved Ets-binding sites within the promoter region of the Fli1 gene and governs Fli1 expression. Importantly, in the absence of Etv2 at midgestation, binding of Etv2 at Ets-binding sites in the Fli1 promoter is replaced by Fli1 protein itself, sustaining expression of Fli1 as well as selective Etv2-regulated endothelial genes to promote endothelial cell survival and vascular integrity. Consistent with this, we report that Fli1 binds to the conserved Ets-binding sites within promoter and enhancer regions of other Etv2-regulated endothelial genes, including Tie2 , to control their expression at and beyond midgestation. Conclusions: We have identified a novel positive feed-forward regulatory loop in which Etv2 activates expression of genes involved in vasculogenesis, including Fli1 . Once the program is activated in early embryos, Fli1 then takes over to sustain the process in the absence of Etv2.

A field experiment was conducted to study the effect of phosphorus (P) deficiency on the mineral nutrition of five modern, high-yielding rice varieties, Purbachi, BR1, BR3, BR14, and BRRIDHAN29. The available P in the soils of different plots of the experimental field varied from 2.8 to 16.4 mg kg−1. Such a wide range of available P in the different plots resulted from differences in the amounts of P (0 to 480 kg ha−1) applied over a period of 8 years during a previous long-term P fertilizer trial in the same field. The relationship among the P, potassium (K), and magnesium (Mg) concentrations in rice straw and grain in a soil P deficiency situation were especially noteworthy. The K concentration in rice straw was negatively correlated with the P concentration (r = −0.42 to −0.60), but in the case of grain the coefficient of correlation between P and K concentration were positive (r = 0.60 to 0.74). There was a strong positive relationship between the P and Mg concentration in grain (r = 0.70 to 0.97). Th...

A field experiment was conducted on an Aerie Haplaquept soil to study the effect of phosphorus (P) deficiency in soil on the P nutrition and yield of five modern varieties of rice, viz., Purbachi, BR1, BR3, BR14, and BR29, popular with the rice farmers of Bangladesh. Soil‐available P in the different plots of the experimental field varied widely, from 2.8 to 16.4 ppm. This plot to plot variation in soil‐available P content resulted from differences in the total amounts (0 to 480 kg ha‐1) of P the plots had received over a period of 8 years in a long‐term P fertilizer trial conducted previously in the same field. Phosphorus deficiency in soil drastically reduced the grain yield of all the rice varieties. In severely P deficient plots, where soil‐available P was around 3 ppm, the yield was less than 1 ton ha‐1 while in plots containing an adequate P level, i.e., >6 ppm, the yield was more than 41 ha‐1. Rice yield increased linearly with an increase in soil P content up to 6 ppm, and the highest gra...

Two popular concepts of soil fertilization, basic cation saturation ratio (BCSR) and sufficiency level of available nutrients (SLAN), were tested on a calcareous soil (Aeric haplaquept) during 1995-1996 at the Bangladesh Rice Research Institute (BRRI) Regional Station Rajshahi using wheat as a test crop. According to BCSR concept the soil was deficient in potassium (K) and according to SLAN concept it was deficient in phosphorus (P), respectively. Potassium dose of 120 kg ha-1 [to attain 2% saturation of total cation exchange capacity (CEC) according to BCSR] along with other two doses (0 and 60 kg K ha-1) and P dose of 50 kg ha-1 (to attain available P at sufficiency level) along with other two doses (0 and 100 kg P ha-1) were compared in a randomized complete block design. The application of 50 kg P ha-1 significantly increased plant height, spikes m-2, grains per spike, grain and straw yields of wheat over 0 kg P ha-1 with or without K but increasing P dose from 50 to 100 kg P ha-1 did not give additional yields. The agronomic parameters and yields were not affected significantly by K application. Similar results were also observed in nutrient content and nutrient uptake. Thus, SLAN concept appeared as an effective tool for fertilizer recommendation for the calcareous soil while BCSR gave no apparent result there.

A laboratory experiment was conducted to evaluate the effects of water and temperature regimes on availability of soil phosphorus (P). Four soil samples with variable levels of available P collected from a long‐term P frequency trial were incubated under two moisture regimes, continuous waterlogging (CWL) and alternate wetting and drying (AWD) at 20°C and 30°C temperature for 90 days. The samples were extracted with 0.5M NaHCO3 for available P determination after incubation. Increase in temperature increased available P content and this increase was more prominent in CWL conditions. Available P in all the soils tested was higher under CWL than AWD conditions, however, the increase in available P due to CWL environment was more pronounced in soils with higher available P.

In this paper a new coding strategy is proposed for discrete Bidirectional Associative Memories (BAMs). This enables the BAM to operate on real-valued inputs and thus extends its applicability to industrial environments where analog patterns are commonly encountered. An example of the application of the new coding method for encoding fuzzy rules is given.

This paper describes a local area network access protocol for integrated voice and data communications in a token passing ring. The transmission scheme is decentralized and suitable for both voice and data services. In this system access delay for voice packets can be bounded to a desired upper limit, even at heavy network load. The quality of speech remains unaffected at any level of offered data load and all voice packets are served within the specified delay limit. The protocol provides equal priority to pure data at normal network load while voice is given priority over pure data at heavy network load. The delay-throughput characteristic of both voice and data traffic has been evaluated at a transmission speed of 10 Mb s -1 and it is shown that the mean delay for voice packets, at heavy data load, can be limited to a value equal to half the threshold token rotation time.

The objectives of the study were the changes in water tension, redox potential, and the pH value of a silty Eutric Gleysol from young river sediments near Mymensingh, the nutrient concentration of soil solution, as well as the supply of nutrients by rain, irrigation and fertilizing, and the uptake of nutrients by rice during the year. In the case of nitrogen, a moderate loss was observed, caused by the leaching of NH4, and significant losses due to volatilisation and denitrification. Fertilized P was enriched in the soil. The leaching of Ca, K, Mg, Na, Cu, Fe, Mn, and Zn was clearly higher than the addition by rain, irrigation and fertilization. It is concluded, that the loss of nutrients was caused by weathering. Wasser- und Nahrstoffdynamik eines Reisbodens in Bangladesh Untersucht wurden die Wasserspannungen, Redoxpotentiale und pH-Werte eines schluffreichen Eutric Gleysol aus jungen Flussedimenten bei Mymensingh, auserdem die Nahrstoffgehalte der Bodenlosung sowie die Nahrstoffzufuhren mit Niederschlag, Bewasserungswasser und Dungung, und die Nahrstoffentzuge durch Reis im Jahreslauf. Beim Stickstoff ergaben sich masige Verluste durch NH4-Auswaschung und starke Verluste durch Volatilisation und Denitrifikation. Gedungtes P wurde im Boden angereichert. Ansonsten lagen die Auswaschungsverluste an Ca, K, Mg, Na, Cu, Fe, Mn und Zn deutlich hoher als die Zufuhren durch Niederschlag, Bewasserung und Dungung, woraus letztlich auf starke Nahrstoff-Freisetzung durch Verwitterung geschlossen werden kann.

Little is known about the roles of galectins, a family of β-galactoside-binding lectins, in myeloid cell differentiation. In the present experiments, we used HL-60 cells as a model of myeloid cell differentiation. The HL-60 cells were differentiated into eosinophil-, monocyte-, and neutrophil-like cells by coculture with sodium butyrate under a mild alkaline condition, phorbol 12-myristate 13-acetate, and dimethyl sulfoxide, respectively. Thus, the expression of galectins in HL-60 cells during differentiation into three different lineages was assessed. Reverse transcriptase-polymerase chain reaction analyses revealed that undifferentiated HL-60 cells expressed galectin-1, -3, -8, -9, and -10 (identical to Charcot Leyden crystal) mRNAs, and galectin-2, -4, and -7 were negligible before and after the differentiations. We failed to detect evident changes in the mRNA levels of galectin-1 and -8 during the diferentiations. However, during the eosinophilic differentiation, galectin-9 mRNA expression was gradually decreased, whereas galectin-10 mRNA expression was increased. During the course of monocytic differentiation, galectin-9 mRNA expression was down-regulated, whereas galectin-3 mRNA expression was up-regulated. Moreover, only galectin-10 mRNA expression was enhanced in the process of neutrophilic differentiation. These changes in galectin expressions were confirmed by Western blot and flow cytometry analyses. It is thus suggested that changes in the expressions of galectin-3, -9, and -10 are potentially important for myeloid cell differentiation into specific lineages.

Galectin-9 expression was examined in 6 human melanoma cell lines. Among them, MM-BP proliferated with colony formation, but MM-RU failed. RT-PCR analysis revealed evident expression of galectin-9 mRNA in MM-BP but not in MM-RU. MM-BP expressed galectin-9 protein both on the surface and in the cytoplasm, whereas MM-RU expressed it only weakly in the cytoplasm. Exogenous galectin-9 induced in vitro both cell aggregation and apoptosis of MM-RU proliferating without colony formation. Association of galectin-9 expression in melanoma cells with prognosis of the patients bearing melanocytic tumors was further examined. Galectin-9 protein was strongly and homogeneously expressed in melanocytic nevi, but down-regulated in melanoma cells especially in metastatic lesions. High galectin-9 expression was inversely correlated with the progression of this disease, suggesting that high galectin-9 expression in primary melanoma lesions links to a better prognosis.

We have shown previously that vascular endothelial growth factor (VEGF) synthesized by the cellular constituents of small vessels per se, viz. endothelial cells and pericytes, participates in the hypoxia-driven proliferation of both cell types (Nomura, M., Yamagishi, S., Harada, S., Hayashi, Y., Yamashima, T., Yamashita, J., Yamamoto, H. (1995) J. Biol. Chem. 270, 28316-28324; Yamagishi, S., Yonekura, H., Yamamoto, Y., Fujimori, H., Sakurai, S., Tanaka, N., and Yamamoto, H. (1999) Lab. Invest. 79, 501-509). In this study, we examined the expression of the recently isolated VEGF gene family members (placenta growth factor (PlGF), VEGF-B, and VEGF-C) in human dermal microvascular endothelial cells and bovine retinal pericytes cultured under various oxygen tensions. Quantitative reverse transcription-polymerase chain reaction analyses demonstrated that the two cell types possess not only VEGF (VEGF-A) mRNA, but also VEGF-B, VEGF-C, and PlGF mRNAs. Among them, only VEGF-A mRNA was induced under hypoxia. Competitive reverse transcription-polymerase chain reaction showed that, under normoxic conditions, the rank order of mRNA content in endothelial cells was PlGFVEGF-BVEGF-CVEGF-A and that mRNA coding for PlGF was expressed at100-fold higher levels than VEGF-A mRNA. In pericytes, the rank order was VEGF-CVEGF-AVEGF-BPlGF, and approximately 7-fold higher levels of VEGF-C mRNA compared with VEGF-A mRNA were noted in this cell type. Furthermore, antisense inhibition of PlGF protein production lowered the endothelial cell synthesis of DNA under hypoxic conditions. The results suggest that these VEGF family members may also take active parts in angiogenesis.

Introduction: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system characterized by multifocal demyelination and axonal loss. In addition to the role of the immune response in targeting oligodendrocytes and myelin in MS lesions, the extensive inflammatory infiltration is likely to lead to free radical production and oxidative damage to both oligodendrocytes/myelin and neurons. In this study we investigated the extent of free radical damage to DNA in postmortem MS spinal cord. Materials and methods: Serial 10 µm tissue sections were cut from 26 fixed and cryoprotected spinal cord blocks from 11 MS cases (age 64 ± 4 years, pmd 13 ± 2 h) and two controls (age 70 ± 9 years, pmd 22 ± 4 h). Lesions were characterized using HE supplied by Dr S Toyokuni, Kyoto), the most abundant modified DNA base. Results: Quantitative analysis revealed a 150% increase in 8OHdG+ cells relative to normal appearing WM and a 400% increase relative to control WM. A positive correlation was found between the level of macrophage infiltration (HLA-DR+) and 8OHdG-expressing cells. Discussion: These results suggest a role for oxidative damage to DNA due to free radicals in the evolution of the MS lesion.

Among galectin family members, galectin-9 was first described as a potent eosinophil chemoattractant derived from Ag-stimulated T cells. In the present study a role of galectin-9 in the interaction between eosinophils and fibroblasts was investigated using a human lung fibroblast cell line, HFL-1. RT-PCR, real-time PCR, and Western blot analyses revealed that both galectin-9 mRNA and protein in HFL-1 cells were up-regulated by IFN-γ stimulation. On the one hand, IL-4, known as a Th2 cytokine, did not affect the galectin-9 expression in HFL-1 cells. We further confirmed that IFN-γ up-regulated the expression of galectin-9 in primary human dermal fibroblasts. Flow cytometric analysis revealed that IFN-γ up-regulated surface galectin-9 expression on HFL-1 cells. Stimulation of HFL-1 cells with IFN-γ up-regulated adhesion of eosinophils, but not neutrophils, to HFL-1 cells. This adherence of eosinophils to HFL-1 cells was inhibited by both lactose and anti-galectin-9 Ab. These findings demonstrate that IFN-γ-induced galectin-9 expression in fibroblasts mediates eosinophil adhesion to the cells, suggesting a crucial role of galectin-9 in IFN-γ-stimulated fibroblasts as a physiological modulator at the inflammatory sites.