Can the adapted arcometer be used to assess the vertebral column in children?

BACKGROUND: The adapted arcometer has been validated for use in adults. However, its suitability for use in children can be questioned given the structural differences present in these populations. OBJECTIVE: To verify the concurrent validity, repeatability, and intra- and inter-reproducibility of the adapted arcometer for the measurement of the angles of thoracic kyphosis and lumbar lordosis in children. METHOD: Forty children were evaluated using both sagittal radiography of the spine and the adapted arcometer. The evaluations using the arcometer were carried out by two trained evaluators on two different days. In the statistical treatment, the intraclass correlation coefficient (ICC), Pearson's product moment correlation, Spearman's rho, the paired t test, and Wilcoxon's test were used (α=.05). RESULTS: A moderate and significant correlation was found between the x-ray and the adapted arcometer regarding thoracic kyphosis, but no correlation was found regarding lumbar lordosis. Repeatability and intra-evaluator reproducibility of the thoracic kyphosis and lumbar lordosis were confirmed, which was not the case of inter-evaluator reproducibility. CONCLUSION: The adapted arcometer can be used to accompany postural alterations in children made by the same evaluator, while its use for diagnostic purposes and continued evaluation by different evaluators cannot be recommended. Further studies with the aim of adapting this instrument for use in children are recommended. .

Rhipicephalus (Boophilus) microplus: expression and characterization of Bm86-CG in Pichia pastoris / Rhipicephalus (Boophilus) microplus: expressão e caracterização da Bm86-CG em Pichia pastoris

The cattle tick Rhipicephalus (Boophilus) microplus is responsible for great economic losses. It is mainly controlled chemically, with limitations regarding development of resistance to the chemicals. Vaccines may help control this parasite, thereby reducing tick pesticide use. In this light, we performed subcloning of the gene of the protein Bm86-GC, the homologue protein that currently forms the basis of vaccines (GavacTM and TickGardPLUS) that have been developed against cattle ticks. The subcloning was done in the pPIC9 expression vector, for transformation in the yeast Pichia pastoris. This protein was characterized by expression of the recombinant Mut+ strain, which expressed greater quantities of protein. The expressed protein (rBm86-CG) was recognized in the Western-blot assay using anti-Gavac, anti-TickGard, anti-larval extract and anti-rBm86-CG polyclonal sera. The serum produced in cattle vaccinated with the antigen CG rBm86 presented high antibody titers and recognized the native protein. The rBm86-GC has potential relevance as an immunogen for vaccine formulation against cattle ticks.

O carrapato-do-boi Rhipicephalus (Boophilus) microplus é responsável por grandes perdas econômicas. Seu controle é principalmente químico e apresenta limitações quanto ao desenvolvimento de resistência aos princípios ativos. As vacinas podem auxiliar no controle deste parasita diminuindo as aplicações de carrapaticidas. Considerando isso, foi realizada a subclonagem do gene da proteína Bm86-CG, proteína homologa a que atualmente é a base das vacinas desenvolvidas (GavacTM e TickGardPLUS) contra o carrapato-do-boi, no vetor de expressão pPIC9, para ser transformado em levedura, Pichia pastoris. Esta proteína foi caracterizada pela expressão da cepa recombinante Mut+ que expressou maior quantidade de proteína. A proteína expressa, rBm86-CG, foi reconhecida no ensaio de Western-blot pelos soros policlonais anti-Gavac, anti-TickGard, anti-Extrato de larva e anti-rBm86-CG. O soro produzido em bovinos vacinados com o antígeno rBm86-CG apresentou altos títulos de anticorpo e reconheceu a proteína nativa. A rBm86-CG possui potencial relevância como imunógeno para formulação vacinal contra o carrapato de bovinos.

A study of the expression of Fas after fluid percussion brain injury in rats / 法医学杂志

OBJECTIVE@#To study the pathologic diagnosis and the injury time estimation in light closed encephalon injury.@*METHODS@#Mice were hurt by fluid percussion, and were killed at 15, 30 min, 1, 3 , 6, 12 h, 1, 4, 7, 14 d respectively after injury. The expression of Fas-L in the cerebral cortex, thalamus, and hippocampi was detected by immunohistochemistry and the results were assessed by image analysis system.@*RESULTS@#It is showed that the expression of Fas-L could be detected in 1 h after injury, and increased significantly in three hours, and it reached apex 12 h after injury, and decreased gradually four days after injury, and returned normal 14 days after injury.@*CONCLUSION@#This research demonstrated that Fas-L mediated apoptosis appeared not only around brain trauma but also in the brain tissue far away from the traumatic area. It indicted that the expression of Fas-L is a useful target for diagnosis of early brain injury; the regularity of Fas-L expression could be used as one of indication to date the time of brain injury.

Expression of Osteoprotegerin and RANK Ligand in Breast Cancer Bone Metastasis

Bone destruction is primarily mediated by osteoclastic bone resorption, and cancer cells stimulate the formation and activation of osteoclasts next to metastatic foci. Accumulating evidences indicate that receptor activator of NF-kB ligand (RANKL) is the ultimate extracellular mediator that stimulates osteoclast differentiation into mature osteoclasts. In contrast, osteoprotegerin (OPG) inhibits osteoclast development. In order to elucidate a mechanism for cancer-induced osteoclastogenesis, cells from a human breast cancer line, MDA-MB-231, were directly co-cultured with ST2, MC3T3-E1, or with primary mouse calvarial cells. Osteoclast-like cells and tartarate resistant acid phosphatase (TRAP) activities were then quantitated. We examined these cell lines and samples from breast cancer by RT-PCR for the expressions of OPG and RANKL mRNA. Compared to controls, co-culture of MDA-MB-231 cells with stromal or osteoblastic cells induced an increase in number of osteoclasts and TRAP activities. MDA-MB-231 cells alone or breast cancer samples did not express RANKL mRNA. However, co-culture of these cancer cells with stromal or osteoblastic cells induced RANKL mRNA expression and decreased OPG mRNA expression. These experiments demonstrate that direct interactions between breast cancer and stromal or osteoblastic cells induce osteoclastogenesis in vitro through modulating RANKL expression.

High extracellular Ca2+ alone stimulates osteoclast formation but inhibits in the presence of other osteoclastogenic factors

High ambient Ca2+ at bone resorption sites have been implicated to play an important role in the regulation of bone remodeling. The present study was performed to clarify the mode of high extracellular Ca2+ (Ca2+e)-induced modulation of osteoclastogenesis and the expression of receptor activator of nuclear factor-kB ligand (RANKL) and osteoprotegerin (OPG), thereby to define its role in osteoclast formation. Mouse bone marrow cells were cocultured with osteoblastic cells in the absence or presence of osteoclastogenic factors such as 1,25-dihydroxyvitaminD3 (1,25-(OH)2vitD3) and macrophage colony-stimulating factor/soluble RANKL. Ca2+ concentration in media (1.8 mM) was adjusted to 3, 5, 7 or 10 mM. Osteoclast formation was confirmed by the appearance of tartrate-resistant acid phosphatase (TRAP)-positive multinuclear cells and the expression of osteoclast phenotypic markers (calcitonin receptor, vitronectin receptor, cathepsin K, matrix metalloproteinase-9, carbonic anhydrase 2). High Ca2+e alone significantly stimulated osteoclast formation in a dose-dependent manner. However, in the presence of highly osteoclastogenic factors, high Ca2+e significantly inhibited osteoclastogenesis. High Ca2+e alone continuously up-regulated RANKL expression while only transiently increased OPG expression. However, in the presence of 1,25-(OH)2vitD3, high Ca2+e did not change the 1,25-(OH)2vitD3- induced RANKL expression while increased OPG expression. Taken together, these findings suggest that high Ca2+e alone increase osteoclastogenesis but inhibit in the presence of other osteoclastogenic factors. In addition, high Ca2+e-induced osteoclastogenesis may be mediated by osteoblasts via up-regulation of RANKL expression. Meanwhile up-regulated OPG might participate in the inhibitory effect of high Ca2+e on 1,25-(OH)2vitD3-induced osteoclastogenesis.

Differential Expressions of Fas and Fas Ligand in Human Placenta

To investigate the expressions of Fas and Fas ligand (FasL) in human placenta, we studied the expressions of Fas and FasL in placenta with RT-PCR, immunoblotting and immunostaining. We observed amplified products of Fas and FasL transcripts, the band of Fas (52 kDa) and multiple bands of FasL (42-52 kDa) in pla-centa. Fas and FasL localized mainly on fetal vessels and on syncytiotrophoblasts respectively. The differential distribution of Fas and FasL in human placenta may reflect intrinsic expressions of them by trophoblasts during differentiation. The increased expression of Fas in trophoblasts may promote apoptosis of placenta in pathologic condition such as preeclampsia.

Study on expression of the glycoprotein in myocardial cell membrane of the rat's early myocardial ischemic / 法医学杂志

OBJECTIVE@#To explore expression of the glycoprotein in early myocardial ischemic.@*METHODS@#The glycoprotein changes occurred at the early acute cardiac ischemic area induced experimentally by ligation of left coronary artery of 32 SD rats. 6 lectins were measured by means of immunohistochemical methods.@*RESULTS@#Positive staining of PNA could be observed in ischemic area at 5 min after ischemia, and the positive area increased with the prolongation of ischemic period. It became the strongest for 2 h and then decreased.@*CONCLUSION@#This experiment proved that myocardial cell membrane in ischemia expressed D-galactose. This may be of some value in forensic medicine practice.

-Melanocyte Stimulating Hormone (MSH) decreases cyclosporine A induced apoptosis in cultured human proximal tubular cells

The pathogenesis of chronic cyclosporine A (CsA) nephrotoxicity has not been elucidated, but apoptosis is thought to play an important role in CsA induced tubular atrophy. Recently Fas-Fas ligand system mediated apoptosis has been frequently reported in many epithelial cells as well as in T lymphocytes. We investigated the ability of CsA to induce apoptosis in cultured human proximal tubular epithelial cells and also the effect of -MSH on them. Fas, Fas ligand, and an intracellular adaptor protein, Fas-associating protein with death domain (FADD) expression, and poly-ADP ribose polymerase (PARP) cleavage were also studied. CsA induced apoptosis in cultured tubular epithelial cells demonstrated by increased number of TUNEL positive cells and it was accompanied by a significant increase in Fas mRNA and Fas ligand protein expressions. FADD and the cleavage product of PARP also increased, indicating the activation of caspase. In -MSH co-treated cells, apoptosis markedly decreased with downregulation of Fas, Fas ligand and FADD expressions and also the cleavage product of PARP. In conclusion, these data suggest that tubular cell apoptosis mediated by Fas system may play a role in tubular atrophy in chronic CsA nephrotoxicity and pretreatment of -MSH may have a some inhibitory effect on CsA induced tubular cell apoptosis.

Trifluoperazine reduces the expression of CD69 in phytohemagglutinin-activated lymphocytes

Trifluoperazine (TFP) is a phenothiazine capable of inhibiting lymphocyte proliferation as well as natural killer cells (NK) and lymphokine-activated killer cells (LAK) cytotoxic activity. CD69 is a surface molecule induced by various mechanisms of cellular activation. In the present work the modulation of CD69 expression by TFP was investigated on PHA-stimulated peripheral blood mononuclear cells and compared to that of CD25 (IL-2 receptor) expression. Determination of surface molecules was performed in an indirect immunofluorescence assay using anti-CD69 or anti-CD25 monoclonal antibodies, and analyzed by flow cytometry. The time course of the expression of these two molecules differed: CD69 expression was already declining at 48 h, whereas CD25 was still increasing at 72 h after stimulation. TFP (10 muM) reduced CD69 expression by 71.8 per cent at 24 h, 68.4 per cent at 48 h and 24 per cent at 72 h following activation. In contrast, the same dose of TFP did not significantly affect CD25 expression at 24 h but showed an inhibitory effect at later times. These results suggest that different activation pathways are involved in the expression of CD25 and CD69.

Turnover and shedding of the Tc-85 surface glycoprotein of Trypanosoma cruzi trypomastigotes

Tc-85 is an 85-kDa surface glycoprotein specific for the trypomastigote stage of Tripanosoma cruzi which has been implicated in the invasion of host cells by the parasite. Tc-85 has a half-life of 3.5-4 h and is synthesized as a 95-kDa precursor. Processing of the 95-kDa precursor is inhibited by N-p-tosyl-L-lysine chloromethyl ketone, p-chloromercuriphenylsulfonic acid, iodoacetamide or N-ethylmaleimide, but not by aprotinin, antipain or phenylmethylsulfonil fluoride. Tc-85, but not the precursor, is rapidly shed into the medium, allowing a correlation between the decrease of Tc-85 in trypomastigotes and its increase in the culture medium. The shedding of Tc-85 was inhibited 50 per cent by 1 muM tunicamycin, but not by 10 muM swainsonine or 10 muM 1-deoxynojirimycin under the experimental conditions employed. This suggests that N-linked oligosaccharides are important for the shedding phenomenon, although it appears that they do not have to be fully processed for shedding to occur.

Biochemical analysis of glycophospholipids from wild-type mouse lymphoma cells and from Thy-1 negative mutants which do not add glycolipid-anchored proteins.

Thy-1 is a major glycophospholipid (GPL)-anchored protein found on the surface of neurons, epithelial cells, fibroblasts and murine T-lymphomas. Biochemical studies were undertaken to determine if murine T-lymphomas contain glycolipids which may be on the path of GPL-anchor biosynthesis. Biosynthetic labeling experiments on Thy-1-positive (wild-type) cells followed by battery of chemical and enzymatic diagnostics on the isolated [3H]mannolipids have, for the first time, led to description of a set of glycolipids which have properties consistent with their being GPL-anchor precursors. Using these results as a guide, major differences have been observed upon analysis of the radiolabeled mannolipids of Thy-1-negative mutants from 7 complementation classes, A-C, E, F, H and I. The biosynthetic lesions in anchor synthesis have been identified in some of the mutants.