A comparative study of leaf epidermal microstructures in the tribe Sorbarieae (Adenostoma: 3 spp., Chamaebatiaria: 1 sp., Sorbaria: 11 spp., Spiraeanthus: 1 sp.) including related genera Gillenia (2 spp.) and Lyonothamnus (2 spp.) was carried out using scanning electron microscopy (SEM) in order to evaluate their significance in taxonomy. The leaves of Adenostoma, Chamaebatiaria, and Spiraeanthus were amphistomatic, whereas Gillenia, Lyonothamnus, and Sorbaria were hypostomatic. The size range of the guard cells is $7.84-48.7{\times}5.86-38.6{\mu}m$; the smallest one was found in Sorbaria tomentosa var. tomentosa ($7.84-11.8{\times}6.84-10.5{\mu}m$), while the largest measured example was Adenostoma fasciculatum var. obtusifolium ($30.3-48.7{\times}18.8-38.6{\mu}m$). Anomocytic stomata complex were the most frequent type (rarely cyclocytic), with usually both anomocytic and actinocytic types occurring in one leaf. On the surfaces, both the adaxial and abaxial anticlinal walls of the subsidiary cells vary (e.g., straight/curved, undulate, sinuate). Four types (unicellular non-glandular trichome, stellate, glandular trichome, pustular glandular trichome) of trichomes are found in the leaves. The epicuticular wax can be divided two types: membraneous platelets (Lyonothamnus) and platelets (Sorbaria arborea var. arborea, S. arborea var. subtomentosa, S. kirilowii, S. tomentosa var. tomentosa, Spiraeanthus schrenkianus). The trichome diversity (in particular, stellate, gland) and the existence of epicuticular wax may have taxonomic significance, although the leaf epidermal micromorphological characteristics do not provide synapomorphy in this tribe. These leaf micromorphological features are most likely better understood in the Sorbarieae when used in conjunction with external morphological characters.
To examine the leaf epidermal microstructure of three genera (Scopolia s.s., Anisodus, AtroPanthe, including Przewalskia as an outgroup) in the genera Scopolia Jacq. s.l., leaves of 10 species (37 specimens) were investigated by the light microscopy (LM) and scanning electron microscopy (SEM). The stomata of studied taxa were 'amphistomatic type' and the size (guard cell) range was $18-64{\times}11-48{\mu}m$. The size of stomata is slightly differed from between the taxa; the smallest size of stomata were found in the monotypic genus, Przewalskia ($24-27{\times}16-17{\mu}m$), on the other hand the largest one was found in Anisodus carniolicoides ($62-64{\times}43-48{\mu}m$). The stomatal complex was mostly anomocytic (in Scopolia s.s., Anisodus taxa : A. luridus, A. carniolicoides, A. acutangulus) and sometimes anisocytic (in Anisodus tanguticus, Przewalskia, Atropanthe). The stomata is mostly crescent in shape, but rarely circular, especially in Przewalskia tangutica. The shapes of epidermal cells are similar in both adaxial and abaxial sides, and mostly undulate/sinuate polygonal anticlinal wall, but rarely arched in Przewalskia tangutica. The epicuticular wax was not well developed in most studied taxa, except Anisodus tanguticus which is well developed cuticular striae around the stomatal complex. The elongate-headed glandular trichomes were found in Scopolia s.s. and Przewalskia. While the taxa of Anisodus and Atropanthe have not any trichomes (i. e., glabrous), except Anisodus luridus, which has simple or sometimes branched (dendritic- type) non-glandular trichome. Finally, the systematic and ecological significance of the leaf micromorphological features (stomata complex, trichome, etc.) in identification and elucidation of Scopolia s.l. including Przewalskia, especially between or within the genera including among the species is also discussed.
This study, consisting of two experiments, was conducted to determine the effects of feeding heat treated protein and mineral complex (HPM) on milk production and composition, and ruminal fermentation of Holstein dairy cows. In in vitro experiment, HPM levels were 0, 0.2, 1 and 2%, and Timothy hay, which was substrate, was milled as 1 mm size, and the effects of HPM on pH, ammonia and VFA were analyzed after incubation times of 0, 6, 12, 24 and 48 h, respectively. The pH and ammonia production were not significantly different between treatments during the incubation. In addition, generally, total VFA and individual VFA were not affected by HPM on 0, 6 and 24 h. While, total VFA and individual VFA were increased in 0.2% and 1% of HPM supplemented treatments, but decreased in 2% of HPM treatment compared with control on 12 h. On 48 h, total VFA and individual VFA were increased in HPM treatments compared to control (P<0.05). However, A/P ratio was not affected by HPM supplementation. Gas production was higher in HPM treatment compared to control on 24 h (P<0.05) and 48 h (P<0.05). In lactating experiment, fourteen lactating Holstein cows were used for 4 months in a cross over experimental design. There were two treatments; no added HPM as a control and 0.2% of HPM added as a test treatment. Daily milk yield (P<0.001), 4% FCM (P<0.001), milk protein (P<0.05) and SNF (solid not fat; P<0.05) were increased in HPM treatment compared to control. While, milk fat, MUN (milk urea nitrogen) and SCC (somatic cell count) were not significantly different between treatments.
Jeong, Il Cheon;Kim, Ji Tae;Hwang, You Sik;Kim, Jung A;Lee, Jae Seung
Clinical and Experimental Pediatrics
/
v.50
no.2
/
pp.178-181
/
2007
Purpose : The renal manifestations of tuberous sclerosis complex (TSC) are remarkably diverse, including polycystic kidney disease, simple renal cysts, renal cell carcinomas, and angiomyolipomas. All of these occur in children as well as adults in TSC. Angiomyolipomas, which can cause spontaneous life-threatening hemorrhages, are by far the most prevalent and the greatest source of morbidity. Here, we will address our experience, adding to the literature on pediatric patients with TSC requiring evaluation and treatment for renal manifestations. Methods : A retrospective analysis was made on 19 patients in whom TSC was diagnosed between May 2001 and Oct. 2005 at Severance Hospital. All patients had clinical diagnoses of TSC as defined by the 1998 tuberous sclerosis complex consensus conference. Results : The patients consisted of 13 boys and 6 girls with a mean age of 7.3 years (range 1 to 22). The renal disease associated with TSC included angiomyolipoma in nine patients (47.4 percent), renal simple cyst in one (5.3 percent), hydronephrosis in one (5.3 percent) patient. Eight patients (42.1 percent) presented with normal kidney contours at abdominal ultrasonography. One patient underwent renal replacement therapy due to chronic renal insufficiency after nephrectomy. Hemorrhage from angiomyolipoma was not detected. Conclusion : In our review of 19 cases of TSC, renal manifestations are reported in 57.9 percent of patients. Asymptomatic angiomyolipoma associated with TSC grow gradually, although severe hemorrhages are rare. So patients with TSC should be followed up with serial computerized tomography or abdominal ultrasonography. And also, renal function should be monitored conservatively.
H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.
Lee, Kyoung-Hee;Lee, Choon-Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yoo, Chul-Gyu
Tuberculosis and Respiratory Diseases
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v.57
no.5
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pp.449-460
/
2004
Background : PS-341 is a novel, highly selective and potent proteasome inhibitor, which showed cytotoxicity against some tumor cells. Its anti-tumor activity has been suggested to be associated with modulation of the expression of apoptosis-associated proteins, such as p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax and Bcl-2. c-Jun N-terminal kinase (JNK) and glycogen synthase kinase-$3{\beta}$ (GSK-$3{\beta}$) are important modulators of apoptosis. However, their role in PS-341-induced apoptosis is unclear. This study was undertaken to elucidate the role of JNK and GSK-$3{\beta}$ in the PS-341-induced apoptosis in lung cancer cells. Method : NCI-H157 and A549 cells were used in the experiments. The cell viability was assayed using the MTT assay and apoptosis was evaluated by proteolysis of PARP. The JNK activity was measured by an in vitro immuno complex kinase assay and by phosphorylation of endogenous c-Jun. The protein expression was evaluated by Western blot analysis. Dominant negative JNK1 (DN-JNK1) and GSK-$3{\beta}$ were overexpressed using plasmid and adenovirus vectors, respectively. Result : PS-341 reduced the cell viability via apoptosis, activated JNK and increased the c-Jun expression. Blocking of the JNK activation by overexpression of DN-JNK1, or pretreatment with SP600125, suppressed the apoptosis induced by PS-341. The activation of caspase 3 was mediated by JNK activation. Blocking of the caspase 3 activation suppressed PS-341-induced apoptosis. PS-341 activated the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, but its blockade enhanced the PS-341-induced cell death via apoptosis. GSK-$3{\beta}$ was inactivated by PS-341 via the PI3K/Akt pathway. Overexpression of constitutively active GSK-$3{\beta}$ enhanced PS-341-induced apoptosis; in contrast, this was suppressed by dominant negative GSK-$3{\beta}$ (DN-GSK-$3{\beta}$). Inactivation of GSK-$3{\beta}$ by pretreatment with lithium chloride or the overexpression of DN-GSK-$3{\beta}$ suppressed both the JNK activation and c-Jun up-regulation induced by PS-341. Conclusion : The JNK/caspase pathway is involved in PS-341-induced apoptosis, which is negatively regulated by the PI3K/Akt-mediated inactivation of GSK-$3{\beta}$ in lung cancer cells.
Background and Objectives: Acute lymphoblastic leukemia (ALL) is a complex genetic disease involving many fusion oncogenes (FO) having prognostic significance. The frequency of various FO can vary in different ethnic groups, with important implications for prognosis, drug selection and treatment outcome. Method: We studied fusion oncogenes in 101 pediatric ALL patients using interphase FISH and RT-PCR, and their associations with clinical features and treatment outcome. Results: Five most common fusion genes i.e. BCR-ABL t (22; 9), TCF3-PBX1 (t 1; 19), ETV6-RUNX1 (t 12; 21), MLL-AF4 (t 4; 11) and SIL-TAL1 (del 1p32) were found in 89/101 (88.1%) patients. Frequency of BCR-ABL was 44.5% (45/101). BCR-ABL positive patients had a significantly lower survival ($43.7{\pm}4.24$ weeks) and higher white cell count as compared to others, except patients with MLL-AF4. The highest relapse-free survival was documented with ETV6-RUNX1 (14.2 months) followed closely by those cases in which no gene was detected (13.100). RFS with BCR-ABL, MLL-AF4, TCF3-PBX1 and SIL-TAL1 was less than 10 months (8.0, 3.6, 5.5 and 8.1 months, respectively). Conclusions: This is the first study from Pakistan correlating molecular markers with disease biology and treatment outcome in pediatric ALL. It revealed the highest reported frequency of BCR-ABL FO in pediatric ALL, associated with poor overall survival. Our data indicate an immediate need for incorporation of tyrosine kinase inhibitors in the treatment of BCR-ABL+ pediatric ALL in this population and the development of facilities for stem cell transplantation.
Background: Cell growth is a balance between cell proliferation and cell death. Insulin-like growth factor-I(IGF-I), which binds IGF-I receptor(IGF-IR), mediates cellular proliferation as a potent mitogen. IGF binding protein-3(IGFBP-3) as a circulating major IGFBP can inhibit or enhance the effects of IGF-I on cellular growth by binding IGFs. Methods: We investigated the expressions of mRNA of IGF-I and IGF-IR by northern blot and phosphorylation of IGF-IR with the treatment of IGF-I by western blot in 3T3 fibroblast cells. The cellular proliferations of 3T3 cells with the treatments of IGF-I were evaluated using $^3H$-thymidine incorporation and MTT assay. Also to observe the effect of IGFBP-3 on cellular proliferation, 3T3 cells were treated with anti-IGFBP-3 and ${\alpha}IR_3$(monoclonal antibody to IGF-IR) alone or in combination. Results: Our results demonstrated that 3T3 cells showed mRNA expressions of IGF-I and IGF-IR and the IGF-I increased phosphorylation of IGF-IR. The treatments of 3T3 cells with IGF-I increased cellular proliferation in 5 % and 1 % seruma-containing media, not in serum-free media. The addition of anti-IGFBP-3 to neutralize IGFBP-3 showed 2-fold increase of cellular proliferation, and also co-incubation of anti-IGFBP-3 and ${\alpha}IR_3$ together showed similar increase of cellular proliferation in 3T3 cells. Interestingly, when the cells were pretreated with ${\alpha}IR_3$ for 4 hr, prior to the simultaneous addition of ${\alpha}IR_3$ and anti-IGFBP-3, anti-IGFBP-3-mediated cellular proliferation was decreased to control level. All of these results suggest that free IGF-I released from IGF-I/IGFBP-3 complex would be involved in the cellular proliferation. Conclusion: IGF-I is a mitogen through the activation of IGF-IR in 3T3 cells, and IGFBP-3 could be a potent inhibitor for IGF-I action by binding IGF-I.
Background : Anti-apoptotic proteins may be involved in tumor development, progression and the response to treatment, Bcl-2 is by far the most studied anti-apoptotic protein. A novel inhibitor of apoptosis, designated survivin, and the heat shock proteins (HSPs) have recently been found in many human cancers. Immunohistochemical methods were used to determine the expression level of survivin, HSP70 and bcl-2 in non-small cell lung cancer (NSCLC) to evaluate their clinical significance. Materials and Methods : Tissue array slides were obtained from 99 surgically resected NSCLCs. Immunohistochemical staining was performed by an immuno-peroxidase technique using an avidin-biotinylated horseradish peroxidase complex. Anti-survivin rabbit polyclonal antibodies, anti-HSP70 mouse monoclonal antibodies and anti-bcl-2 mouse monoclonal antibodies were used as the primary antibodies. Results : Positive staining of survivin was detected in 33.3% of the cases. Survivin positivity is associated with to females and recurrence. A nonstatistically significant trend toward increased survivin expression was observed in non-smokers, and its expression inversely correlated with the number of cigarettes smoked in smokers. HSP70 was detected in 84.8% but this did not correlated with the clinicopathologic characteristics. Bcl-2 was detected in 18.2% and its expression correlated to tumor recurrence. No significant difference in the median survival time was noted in a comparison of all cases with survivin expression and those without. There was no association between HSP70 or bcl-2 expression and survival. Conclusion : Survivin expression was significantly associated with females and tumor recurrence. In addition its expression was inversely associated with the number of cigarettes smoked. However, HSP70 and bcl-2 expression were not associated with the clinical parameters or survival. This suggests that measuring the survivin levels may be useful in identifying patients at high risk for disease recurrence. Therefore, survivin might be a new diagnostic/therapeutic target in cancer.
Cyclin I plays a pivotal role in the regulation of G1-S transition and could consequently be a deregulated molecule in tumors. The activity of the cdk2-cyclin E complex is increased by degradation of cdk inhibitor p27kip1. Little is known about the expression and prognostic significance of cyclin E and p27 in non-small cell lung cancer(NSCLC). Material and Method: The expression of cyclin E and p27 in eighty-one cases of resected stage I NSCLC tissues and its relation to major clinico-pathological factors, including histology, differentiation, size of tumor, pleural invasion and survival rate were studied and analyzed. Immunohistochemical analysis with monoclonal antibodies specific for cyclin E and p27 were performed by ABC method. Result: Expression rates of cyclin E and p27 in stage I NSCLC tissues were 29.6% and 28.4% respectively. Cyclin E was expressed higher in cases of pleural invasion(p=0.04), and p27 was expressed higher in diameter of tumor less than 3cm(p=0.015). The 5-years survival rate was lower in cases of Positive expression of cyclin E than in cases of negative expression of cyclin E(44.4% vs 68.2%, p=0.015), and the 5-years survival rate was 72.2% in positive expression of p27 and 56.2% in negative expression of p27(p=0.09). The 5-years survival rate was higher in negative expression of cyclin E and positive expression of p27 than in cases of positive expression of cyclin I and negative expression of p27 (73.5% vs 36.3%, p=0.0029). In multivariate analysis, expression of cyclin I was an unfavorable prognostic factor(RR=3.578, p=0.006) and p27 was a favorable prognostic factor(RR=0.183, p=0.019) independently. Conclusion: Cyclin E and p27 may play a pivotal role for the biological behavior of stage I NSCLC, so that the expressions of cyclin I and p27 nay be new prognostic markers.
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