Production and Evaluation of Toxoplasma gondii Recombinant Surface Antigen 1 (SAG1) for Serodiagnosis of Acute and Chronic Toxoplasma Infection in Human Sera

Background

The assays currently available for the detection of specific anti-Toxoplasma antibodies may vary in their abilities to detect serum immunoglobulins, due to the Lack of a purified standardized antigen. The aim of this study was evaluation the recombinant Toxoplasma gondii SAG1 antigen for the serodiagnosis of acute and chronic toxoplasmosis.

Methods

This study describes an ELISA using recombinant SAG1 for detection of IgM and IgG antibodies against Toxoplasma gondii in human sera. Genomic DNA of T. gondii (RH Strain) was isolated and PCR reaction was performed. Recovered DNA was cloned into PTZ57R cloning vector. The recombinant plasmid was detected by restriction analysis. The SAG1 gene was subcloned in the pET- 28a expression vector. Protein production was then induced with 1 mM isopropyl-D – thiogalactopyranoside (IPTG). A total of 204 sera were tested using a commercial IgG and IgM ELISA kit (Trinity, USA) as gold standard prior to testing them with the recombinant antigen.

Results

Tested sera were divided into the following groups:(a) The 74 T. gondii IgG positive (b) 70 T.gondii IgM positive (c) 60 sera who had no serological evidence of toxoplasmosis as negative sera.To determine the specificity of the test, we used other parasitic diseases including echinococusis (N=5), malaria (N=14), leishmaniasis (N=7),fasciolasis (N=4), sterengyloidiasis (N=1). Sensitivity and specificity of the generated recombinant IgG ELISA in comparison with commercial ELISA (Com ELISA) were 93% and 95%, and the sensitivity and specificity of the generated recombinant IgM ELISA were 87% and 95% respectively.

Conclusion

The results acquired here show that this antigen is useful for diagnostic purposes and could be replaced by lysed, whole cell antigens for diagnosis of chronic toxoplasmosis.     

Storage of pools of six and eight platelet concentrates

Platelet concentrates (PCs) prepared from units of whole blood are routinely stored singly at 20 to 24 degrees C and pooled prior to transfusion. Studies have been conducted to evaluate the in vitro properties of pools of six (n = 19) and eight (n = 17) ABO-identical PCs after storage, with comparative studies involving single units (n = 33). The pools were prepared using the sterile connecting device. One- day-old and 3-day-old PCs were pooled and stored for a total of 5 days in a container system consisting of two 1000-mL polyolefin containers. The pooled platelet suspension was divided approximately equally between the two containers. The platelet count was reduced by less than 5 percent during storage of the pools, which is similar to the reduction found with storage of control units of single PCs. The volume loss due to pooling was 9.6 +/− 1.9 percent (mean +/− 1 SD). The pH of the PC pools was approximately 7.0 after 5 days of storage, with no pool having a pH below 6.2. In vitro platelet properties, such as morphology score, extent of shape change induced by ADP, total ATP, aggregation response to ADP and collagen, response to hypotonic stress, lactate dehydrogenase discharge, and beta-thromboglobulin release, were similar for pools and control single PCs. In addition, comparable low levels of thymidine uptake were detected in the mononuclear leukocyte fraction of pooled and unpooled PCs that were stored for 5 days at 20 to 24 degrees C, which indicates that the mixing of lymphocytes in the pool did not stimulate in vitro immunologic reactions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cardiac β-adrenoceptor sensitivity and Parkinson''s disease

Certain clinical manifestations of Parkinson's disease (PD) (speech or/and balance disturbances) are not linked to brain dopamine deficiency. The purpose of the present study was to search for a possible relationship between those so-called "non-dopamine-dependent" extrapyramidal manifestations and the sensitivity of cardiac beta-adrenoceptors. Fourteen patients aged 51 to 69 were included in the study after having given their informed consent. Any factor or pathology susceptible to modify receptor sensitivity entailed exclusion. In the absence of a reference model for measuring the reactivity of central beta-adrenoceptors, a computation of the isoprenalin dose necessary to increase the resting heart rate by 20 bpm was used as an index for beta-adrenergic system reactivity. In addition to that test, other parameters were recorded: disease duration, motor status scale (Columbia), some cognitive functions (MMS and image differed recall). The cardiac beta-receptor decrease in reactivity to isoproterenol is correlated to PD duration (r = 0.8, P less than 0.001). Conversely, the sensitivity of these receptors appeared to be unrelated to the extrapyramidal severity of the disease, hence to the degree of the so-called "non dopamine-dependent" disturbances. Furthermore, such results raise the meaning of the impairment of peripheral aminergic receptors in the cognitive disturbances linked to ageing and/or PD.     

同种异体胚胎神经干细胞移植修复脊髓损伤的时效性观察

目的:观察同种异体胚胎神经干细胞移植对脊髓损伤大鼠双后肢运动功能的修复作用,并分析其移植时效性。方法:实验于2003-07/08在大连医科大学分子生物学实验室和大连理工大学环境与生命学院生物医学实验室完成。①取孕14￣16d的大白鼠1只,引颈处死,剖腹,取出胎鼠,钳取大脑皮质及皮质下脑室旁的脑组织,体外培养大鼠胚胎神经干细胞。②将30只成年SD大鼠按随机数字表法分为3组,即损伤对照组、早期移植组和延期移植组,每组10只。3组大鼠均制作脊髓横断损伤模型,造成大鼠下肢瘫痪,早期移植组、延期移植组大鼠分别在伤后3d及3周移植胎鼠脑组织神经干细胞。观察移植后大鼠双后肢的运动功能恢复情况,通过胶质原纤维酸性蛋白及神经元特异性烯醇化酶免疫组化染色法观察各组大鼠的脊髓组织形态学变化,胶质原纤维酸性蛋白是星形胶质细胞特异性标志蛋白,神经元特异性烯醇化酶是神经元特异性蛋白。结果:脊髓损伤建模实验纳入大鼠30只,全部进入结果分析,无脱失。①各组大鼠神经干细胞移植后双后肢的功能恢复情况:早期移植组和延期移植组大鼠双后肢的运动功能均有明显改善,但早期移植组表现尤为显著。早期移植组和延期移植组细胞移植后五六天即可见瘫痪大鼠的后肢肌力开始恢复,二三周后可出现爬行,4周后后肢活动活跃;损伤对照组大鼠的瘫痪肢体无任何恢复。②移植4周后早期移植组大鼠移植区脊髓组织的形态学变化:脊髓移植区肉眼可见有增生的组织充填,镜下有大量新生的细胞,表现为神经元和神经胶质细胞阳性染色(胶质原纤维酸性蛋白( ),神经元特异性烯醇化酶( ))。结论:神经干细胞移植对脊髓横断大鼠运动功能恢复有促进作用,早期移植疗效更好。     

CD79a: a novel marker for B-cell neoplasms in routinely processed tissue samples

The CD79 molecule, comprising two polypeptide chains, mb-1 (CD79a) and B29 (CD79b), is physically associated in the B-cell membrane with immunoglobulin. It transmits a signal after antigen binding and may, therefore, be considered the B cell equivalent of CD3. It appears before the pre-B-cell stage, and the mb-1 (CD79a) chain can still be present at the plasma cell stage. In this report, we describe a new anti-CD79a monoclonal antibody, JCB117, which reacts with human B cells in paraffin embedded tissue sections, including decalcified bone marrow trephines. When tested on a total of 454 paraffin embedded tissue biopsies, gathered from a number of different institutions, it reacted with the great majority (97%) of B-cell neoplasms, covering the full range of B-cell maturation, including 10 of 20 cases of myeloma/plasmacytoma. It is of interest that the antibody labels precursor B-cell acute lymphoblastic leukemia samples, making it the most reliable B-cell marker detectable in paraffin-embedded specimens in this disorder. All neoplasms of T cell or nonlymphoid origin were negative, indicating that antibody JCB117 may be of value to diagnostic histopathologists for the identification of B-cell neoplasms of all maturation stages.     

Quality of life in mothers of children with psoriasis

Background

Patients' and their mothers' quality of life is severely affected by childhood psoriasis. Almost all children have a chronic illness that lasts until adulthood, which puts them at risk for lifelong difficulties like stigma, psychiatric comorbidity, and suicide.

Objective

Evaluation of the effects of childhood psoriasis on the mothers' quality of life was the project's primary objective.

Subjects and Methods

100 mothers of children with various kinds of psoriasis participated in the study. The Family Dermatology Life Quality Index (FDLQI) was used to evaluate the mothers' quality of life.

Results

The mother's FDLQI score was between 3 and 25, with a mean of 13. In terms of how the FDLQI was interpreted, 8 moms had an incredibly enormous impact, 63 mothers had a very significant impact, 26 mothers had a moderate impact, and 3 mothers had a modest impact. We discovered a substantial direct link between the mother's FDLQI and the children's PASI scores. Furthermore, we discovered that scalp and pustular psoriasis had the highest FDLQI scores, indicating a poor quality of life.

Conclusion

Both the quality of life for affected children and their cares may be negatively impacted by childhood psoriasis. Age of the children, PASI score, and kind of psoriasis can all have an impact on how psoriasis in childhood affects the mother.