Plasma as a scaffold for regeneration of neural precursor cells after transplantation into rats with spinal cord injury

The present study investigated whether plasma could be useful as a scaffold for cell transplantation in rats with spinal cord injury (SCI). Transplantation of cells with plasma promoted the recovery of SCI-induced motor dysfunction. Immunohistochemical analysis revealed that the grafted cells had differentiated into the neural lineage. When dissociated neural precursor cells were cultured with plasma, extensive neurite outgrowth was observed along with increased expression of p35 and NF68. Neural markers were also expressed by the cultured cells. Culture with plasma reduced thymidine incorporation, but promoted cell growth and increased the RNA contents. These findings suggest that the cells underwent differentiation into neurons in the presence of plasma. In conclusion, plasma could be a promising scaffold for cell transplantation therapy.     

Effects of sepsis on the neuromuscular blocking actions of d-tubocurarine on rat adductor and abductor laryngeal muscles

Purpose

We evaluated the effects of sepsis on the neuromuscular blocking actions of d-tubocurarine (dTc) in the lateral cricoarytenoid (LCA) and posterior cricoarytenoid (PCA) muscles, an adductor muscle and an abductor muscle of the vocal cords, respectively, in vitro.

Methods

Sepsis was induced in rats by cecal ligation and puncture (CLP) to elicit panperitonitis. Electromyograms (EMGs) and endplate potentials (EPPs) were recorded from the LCA and PCA muscles of CLP-operated septic rats and sham-operated nonseptic rats, using extracellular and intracellular microelectrodes, respectively.

Results

EMG and EPP (amplitude and quantum content) were depressed by dTc, but the dTc-induced neuromuscular blocking effects were attenuated by sepsis. The suppressive effects of dTc on EMG and EPP (amplitude and quantum content) were less intense in the LCA muscle than in the PCA muscle under both sepsis and nonsepsis conditions.

Conclusion

Our study shows that sepsis has a depressive effect on dTc-induced neuromuscular blocking actions at both the adductor and abductor muscles of vocal cords in the larynx.     

Salt-losing nephrogenic diabetes insipidus caused by fetal exposure to angiotensin receptor blocker

The administration of angiotensin-converting enzyme inhibitors (ACEIs) or angiotensin type 1 receptor blockers (ARBs) to pregnant women has been reported to cause ACEI/ARB fetopathy, including oligohydramios, pulmonary hypoplasia, renal insufficiency, limb contracture, and fetal hypotension in the child. Most of the patients die or develop end-stage renal failure during the neonatal period. The long-term prognosis of renal dysfunctions of patients with ARB fetopathy has not been reported. We report two pediatric cases, a 6- and 2-year-old boy, respectively, with ARB fetopathy whose renal functions were thoroughly evaluated after recovery from neonatal renal failure. Both patients showed (1) mildly decreased glomerular filtration rate, (2) no significant proximal tubular dysfunctions, and (3) salt-losing nephrogenic diabetes insipidus, while the excretion of arginine vasopressin and urine level of cyclic AMP were increased. The data on these two patients indicate that the administration of ARB to the fetus profoundly impairs the urine concentrating ability, probably due to papillary atrophy and the disturbed formation of the osmotic gradient in the medulla, which have been confirmed in neonatal rats administered with ACEIs or ARBs. ACEIs/ARBs must not be administered to pregnant women.     

1～3岁幼儿面孔表情认知发展特征研究

目的探讨1～3岁幼儿对5种面孔表情的认知反应发展特征。方法选择2006年11月至2008年11月,在中山大学附属第三医院和佛山市妇幼保健院常规保健的1～3岁幼儿180例作为观察对象,让幼儿观看电脑屏幕上呈现的5种标准面孔表情图,记录、编码和分析幼儿对面孔表情图的视觉注意、自身情绪和行为反应。结果1～2岁组幼儿对5种不同表情图的初次注视时间比较,差异无显著意义（P〉O．05）,2～3岁组幼儿对5种不同表情图的初次注视时间比较,差异无显著意义（P〈O．01）。幼儿对5种表情图的回看次数和组别效应均不显著。1～1.5岁组幼儿对5种表情的初次注视时间和总注视时间短于其他两组（P〈O．01）。各组幼儿对5种不同表情图的积极和消极情绪得分比较,差异有显著意义（P〈0．05）。各组幼儿回看母亲、手指指向行为发生率比较,差异无显著意义（P〉0．05）,模仿表情行为发生率和语言表达能力比较,差异有显著意义（P〈o．001）。结论1岁以后,幼儿基本能感知和识别不同情绪意义的面孔表情,并开始逐渐对表情进行分类和概念化,但很粗浅。     

Noteworthy Cardiovascular Involvement with Sporadic Late-onset Nemaline Myopathy

Cardiac involvement has recently been the focus of sporadic late-onset nemaline myopathy (SLONM). However, right ventricular failure and pulmonary hypertension, in addition to repetitive cardiac arrest, are noteworthy characteristics of SLONM. We herein report a 66-year-old woman with SLONM whose main symptoms were cardiac arrest, right ventricular failure, and pulmonary hypertension. Despite permanent pacemaker replacement, cardiac arrest occurred repetitively, and even with continuous positive airway pressure, right ventricular failure and pulmonary hypertension persisted. The patient was finally diagnosed with SLONM by a muscle biopsy. Our case suggests the possibility of cardiovascular involvement in SLONM, especially right ventricular failure and pulmonary hypertension.     

Identification of Secreted Antigen 3 from Babesia gibsoni

A cDNA expression library of Babesia gibsoni was screened with the serum collected from a dog experimentally infected with B. gibsoni. A novel antigen sharing homology with secreted antigen 1 of B. gibsoni was isolated. The genomic analysis indicated that the BgSA3 gene exists as multicopies in the genome of B. gibsoni. The putative peptide encoded by the BgSA3 gene showed some characteristics of secreted proteins. The serum raised in mice immunized with the recombinant BgSA3 expressed in Escherichia coli could recognize a native parasite protein with a molecular mass of 70 kDa. Moreover, a sandwich enzyme-linked immunosorbent assay with anti-BgSA3 antibodies could detect the circulating BgSA3 in the blood plasma of dogs experimentally infected with B. gibsoni. The identification of BgSA3 provided a useful target for the development of a diagnostic test for detecting specific antibodies and circulating antigens.Babesia canis and Babesia gibsoni are recognized as the two species that cause canine babesiosis, a clinically significant hemolytic disease of dogs. Individual Babesia species are typically classified by the size and morphological appearance of the intraerythrocytic forms, often referred to as piroplasms. B. gibsoni is considered to be a small babesial parasite with round or oval intraerythrocytic piroplasms. The infection caused by B. gibsoni is characterized by serious clinical problems, including remittent fever, progressive anemia, hemoglobinuria, and sometimes death (4, 6). This disease is epidemic in many areas of Asia, North America, and Northern and Eastern Africa but rarely so in Europe (4, 12, 18).For the diagnosis of this disease, several serological methods have been developed, such as indirect fluorescent antibody tests (IFAT) (24), enzyme-linked immunosorbent assays (ELISAs) (13, 22), and immunochromatographic tests (14). However, the detection of antibodies is unreliable for determining the infection status of dogs because the titer of the antibodies against the parasites can remain very high for a long time, even when the parasites have been completely eliminated. However, the secreted proteins, which include a broad variety of antigens released by the parasites and circulating in the bloodstream of the hosts during the asexual stage, can be used as diagnostic targets in antigen detection tests in order to avoid such problems. In an attempt to identify these circulating antigens, a method to screen a cDNA library was designed in a previous study (13). Several antigen candidates were isolated, and one of them, named secreted antigen 1 of B. gibsoni (BgSA1), was identified and evaluated as a useful antigen for further serologically diagnostic tests (13). Circulating BgSA1 could be detected in the plasma of a dog infected with B. gibsoni. In addition, the ELISA using the recombinant BgSA1 expressed in Escherichia coli showed advantages in sensitivity and specificity when it was used in field samples.In this study, we describe the identification of another member of secreted antigens, which share homology with BgSA1, here designated as secreted antigen 3 of B. gibsoni (BgSA3). The gene encoding BgSA3 was isolated from a cDNA library by immunoscreening with serum from a dog experimentally infected with B. gibsoni. As expected, the native BgSA3 also circulated in the bloodstream of the dogs infected with B. gibsoni. Our data indicate that BgSA3 could be useful as an antigenic marker of active B. gibsoni infection.     

Cell labeling for magnetic resonance imaging with the T1 agent manganese chloride

There is growing interest in using MRI to track cellular migration. To date, most work in this area has been performed using ultra-small particles of iron oxide. Immune cells are difficult to label with iron oxide particles. The ability of adoptively infused tumor specific T cells and N cells to traffic to the tumor microenvironment may be a critical determinant of their therapeutic efficacy. We tested the hypothesis that lymphocytes and B cells would label with MnCl2 to a level that would allow their detection by T1-weighted MRI. Significant signal enhancement was observed in human lymphocytes after a 1 h incubation with 0.05-1.0 mM MnCl2. A flow cytometry-based evaluation using propidium iodide and Annexin V staining showed that lymphocytes did not undergo apoptosis or necrosis immediately after and 24 h following a 1 h incubation with up to 1.0 mM MnCl2. Importantly, NK cells and cytotoxic T cells maintained their in vitro killing capacity after being incubated with up to 0.5 mM MnCl2. This is the first report to describe the use of MnCl2 to label lymphocytes. Our data suggests MnCl2 might be an alternative to iron oxide cell labeling for MRI-based cell migration studies.     

Clodronate inhibits PGE(2) production in compressed periodontal ligament cells

Periodontal ligament (PDL) cells play an essential role in orthodontic tooth movement. We recently reported that clodronate, a non-N-containing bisphosphonate, strongly inhibited tooth movement in rats, and thus could be a useful adjunct for orthodontic treatment. However, it is not clear how clodronate affects the responses of PDL cells to orthodontic force. In this study, we hypothesized that clodronate prevents the mechanical stress-induced production of prostaglandin E(2) (PGE(2)), interleukin-1beta (IL-1beta), and nitric oxide (NO) in human PDL cells. A compressive stimulus caused a striking increase in PGE(2) production, while the responses of IL-1beta and NO were less marked. Clodronate concentration-dependently inhibited the stress-induced production of PGE(2). Clodronate also strongly inhibited stress-induced gene expression for COX-2 and RANKL. These results suggest that the inhibitory effects of clodronate on tooth movement and osteoclasts may be due, at least in part, to the inhibition of COX-2-dependent PGE(2) production and RANKL expression in PDL cells.