Immunohistochemical localization of tissue factor pathway inhibitor-1 (TFPI-1), a Kunitz proteinase inhibitor, in Alzheimer''s disease

Senile plaques in Alzheimer's disease (AD) are composed principally of Aβ, a 4 kDa fragment of the amyloid precursor protein (APP). Longer forms of APP which contain a Kunitz proteinase inhibitor (KPI) domain are elevated in aged and in AD brains. Tissue factor pathway inhibitor-1 (TFPI) contains three tandem KPI domains and has been well characterized for its role as a natural anticoagulant in the extrinsic coagulation pathway. Functionally, the first two KPI domains of TFPI bind and inhibit the activity of factor Xa and VIIa respectively. In addition, TFPI and APP-KPI share a common clearance mechanism through the low density lipoprotein receptor-related protein (LRP). As part of an ongoing study of the role of KPI-containing proteins in AD, the current study examines TFPI localization in the brain. We report here that TFPI is immunohistochemically localized to microglia in both AD and non-AD individuals and is localized to some senile plaques in AD. Western blot analyses indicate that the amount of TFPI is elevated in frontal cortex samples from AD brains. We propose that TFPI may play a cell specific role in proteinase regulation in the brain.     

Loss of lipopolysaccharide receptor CD14 from the surface of human macrophage-like cells mediated by Porphyromonas gingivalis outer membrane vesicles

Porphyromonas gingivalis, the major etiologic agent of chronic periodontitis, produces a broad spectrum of virulence factors, including outer membrane vesicles. In this study, we investigated the capacity of P. gingivalis vesicles to promote the shedding or cleavage of the lipopolysaccharide (LPS) receptor CD14 from the surface of human U937 macrophage-like cells. SDS-PAGE/Western immunoblotting analysis of gingival crevicular fluid samples from patients affected by moderate or advanced periodontitis revealed the presence of soluble CD14 and CD14 fragments, thus supporting the hypothesis of an in vivo shedding and cleavage of CD14 receptors. Flow cytometry analysis of macrophage-like cells treated with a vesicle-containing culture supernatant of P. gingivalis showed a significant decrease in the binding of anti-human CD14 to the cell surface. However, no accumulation of soluble CD14 or immunoreactive CD14 fragments in the assay supernatant could be demonstrated by ELISA. Treatment of macrophage-like cells with various concentrations of P. gingivalis vesicles substantially suppressed TNF-alpha production triggered by Escherichia coli LPS. This suppressive effect was much less important using heat-treated vesicles or in the presence of leupeptin, a gingipain inhibitor, during the treatment. Recombinant human CD14 receptors were found to be susceptible to proteolytic degradation by P. gingivalis vesicles. A purified Arg-gingipain preparation produced much more degradation than a Lys-gingipain preparation. This study provides evidence that P. gingivalis outer membrane vesicles contribute to the loss of membrane-bound CD14 receptors and that gingipains degrade this LPS receptor. Such a phenomenon, which results in an hyporesponsiveness of macrophages to LPS stimulation, may contribute to an increased capacity of P. gingivalis, and other periodontopathogens, to evade the host immune system mechanisms.     

Enzyme activities associated with the demyelinating phase of canine distemper

Summary Canine distemper, a naturally occurring viral disease of dogs which often terminates in parainfectious demyelination, was used as a model to study the role of acid proteinase, neutral proteinase and beta-glucuronidase in demyelination. These enzymes were higher in cerebella of dogs with distemper-associated demyelination than in age-matched controls. The highest elevations corresponded with the most severely demyelinated cerebella. The source of the increased enzymes activities was apparently unrelated to the lymphocytes present in areas of demyelination.The direct effect of distemper virus and serum on these enzymes was tested in canine glial monolayers. Virus infection resulted in lower enzyme activities in cells concomitant with the appearance of cellular lesions. There was a relative increase of beta glucuronidase activity in the media suggesting that distemper virus released pre-formed lysosomal enzymes. Serum which was obtained from dogs with distemper-associated demyelination and had previously demyelinated cerebellar explants, also decreased activities of all 3 enzymesin vitro.The 3 enzymes were measured in gerbil brains at various time intervals following unilateral cerebral infarction to determine if processes other than demyelination also caused these enzymes to be increased. Uncomplicated ischemic necrosis (24 h post infarction) did not alter the activities of these enzymes. Invasion of macrophages to ingest and digest necrotic tissue 10 days after infarction resulted in greatly increased acid proteinase and beta-glucuronidase, but unchanged neutral proteinase, activities.It was concluded that the increased activities of acid proteinase and beta-glucuronidase in demyelinated tissue probably are derived from macrophages ingesting damaged tissue. Neutral proteinase may be more specifically involved in the demyelinating process since this is partially located within myelin and can degrade the basic protein of myelin.Supported in part by Research grant Nos. GM 1052 and Al-09022 from National Institutes of Health Service, U.S. Public Health Service.     

Peri-operative elastase-α-1 proteinase inhibitor in patients with postoperative intervertebral discitis

Summary In a prospective study of 251 patients operated upon for lumbar disc herniation it has been investigated whether the preoperative and early postoperative values of Elastase--1 Proteinase Inhibitor (EPI) — an indicator of inflammatory processes — and C-reactive Protein (CRP) — a well known predictor of some postoperative complications — were correlated to the later development of discitis. Postoperatively discitis developed in 14 patients. A randomly choosen group of 15 complication-free patients out of the total of 251 cases was used as control group.Elevated EPI plasma values, especially in the pre-operative and first postoperative days, turned out to be significantly related to the likelyhood of later discitis development, but no such relation for the CRP plasma values could be established. Thus and early prediction of patients at risk for this complication seems to be possible by pre- and postoperative measurement of EPI. It could be justified —but its usefulness has yet to be proven — to give antibiotics prophylactically and other anti-inflammatory medication in patients with elevated pre- and postoperative EPI values.     

The secretion of aspartyl proteinase,a virulence enzyme,by isolates of Candida albicans from the oral cavity of HIV-infected subjects

Prevalence, serotype and in vitro secretion of aspartyl proteinase, a virulence enzyme, were studied in Candida isolates from the oral cavity of 337 HIV-infected subjects. Controls were 95 age-sex-matched HIV- (seronegative) subjects, belonging to either HIV-risk categories (47) or to the normal, general population (48). Fungi were isolated from 155 HIV+ subjects. C. albicans was the most prevalent species (85.8% of all isolates). 94.6% of C. albicans isolates were serotype A and all were agglutinated by a monoclonal antibody (AF1) directed against a major mannoprotein immunogen of the candidal cell wall, confirming previous results with C. albicans isolates from non-immunodeficient subjects. With regard to the stage of HIV infection, there were no statistically significant differences in the incidence of oral Candida carriage between asymptomatic (stage II) HIV+ and HIV- subjects, and between stage II and lymphadenopathic (stage III) individuals. Also, the low (3.8%) incidence of oral candidiasis in the subjects of the latter stage was insignificant with respect to stage II subjects. However, the incidence of C. albicans in stage IV (AIDS) subjects (46.8%) was significantly higher than in all other subjects, and in almost all cases, fungal isolation was accompanied by oral thrush and lower CD4+ lymphocyte counts (< 400 × 10°/L).All isolates of C. albicans were proteolytic in vitro, as assessed by scoring the proteinase activity on BSA agar and monitoring the secreted proteinase antigen by a highly sensitive (1 ng) and specific immunoenzymatic assay. However, by both methods, the isolates from subjects at stages III and IV of infection produced more secretory proteinase than the isolates from either HIV+ asymptomatic subjects or HIV- controls. The differences could not be attributed to particular culture media or source of Candida isolation (carriage versus active infection). Thus, the isolates of C. albicans from advanced HIV infection are serologically similar but more proteolytic than the isolates from earlier stages of HIV infection or those from HIV-uninfected subjects. The apparently higher virulence of C. albicans from AIDS subjects may represent a co-factor in determining and/or aggravating oral candidiasis in these patients.     

酸枣仁汤合小柴胡汤治疗甲状腺功能亢进症50例

目的:观察酸枣仁汤合小柴胡汤治疗甲状腺功能亢进症的临床疗效。方法:选取甲状腺功能亢进症患者100例,按照随机平行分组法分为治疗组与对照组各50例,对照组给予西药甲硫咪唑治疗,治疗组给予中药酸枣仁汤合小柴胡汤治疗,两组疗程均为8周。结果:治疗8周后,两组主要中医证候及血清TT3、TT4、FT3、FT4、TSH水平均明显改善(P0.05),且治疗组均优于对照组(P均0.05)。结论:酸枣仁汤合小柴胡汤治疗甲状腺功能亢进症疗效显著。     

Impact of proteolytic enzymes in colorectal cancer development and progression

Tumor invasion and metastasis is a highly complicated, multi-step phenomenon. In the complex event of tumor progression, tumor cells interact with basement membrane and extracellular matrix components. Proteolytic enzymes (proteinases) are involved in the degradation of extracellular matrix, but also in cancer invasion and metastasis. The four categories of proteinases (cysteine-, serine-, aspartic-, and metalloproteinases) are named and classified according to the essential catalytic component in their active site. We and others have shown that proteolytic enzymes play a major role not only in colorectal cancer (CRC) invasion and metastasis, but also in malignant transformation of precancerous lesions into cancer. Tissue and serum-plasma antigen concentrations of proteinases might be of great value in identifying patients with poor prognosis in CRC. Our results, in concordance with others indicate the potential tumor marker impact of proteinases for the early diagnosis of CRC. In addition, proteinases may also serve as potential target molecules for therapeutic agents.     

The effect of Dodonaea viscosa var. angustifolia on Candida albicans proteinase and phospholipase production and adherence to oral epithelial cells

Aim of the study

This study investigated effect of a crude extract of Dodonaea viscosa on the proteinase and phospholipase production and adherence to epithelial cells by Candida albicans isolated from HIV positive and HIV negative patients.

Materials and methods

Twenty Candida albicans strains isolated from HIV positive and 20 from HIV negative patients were investigated. The isolates were exposed to subinhibitory concentration of crude plant extract and adherence, proteinase and phospholipase production were assessed. The results were analysed using Student's t-test and a two-way ANOVA.

Results

Dodonaea viscosa var. angustifolia inhibited the adherence of Candida albicans to oral epithelial cells (p = <0.01) but no significant effect of the plant extract on proteinase and phospholipase production was observed. Results from Candida albicans strains isolated from HIV positive and HIV negative patients were similar.

Conclusions

Dodonaea viscosa var. angustifolia inhibited the adherence of Candida albicans to oral epithelial cells, which is the initial step of colonization in the infection process. This plant has a therapeutic potential at subinhibitory concentration.     

以肉苁蓉为主要原料的软胶囊抗疲劳抗辐射实验研究

目的探讨以内苁蓉、刺五加、大枣为主要原料制成的软胶囊抗疲劳和抗辐射作用。方法设立3个实验组，每天分别以0．18、0．36和1．08g／kg．bw剂量的软胶囊给小鼠灌胃一次，同时设立空白对照组，连续灌胃30d后，测定小鼠的负重游泳时间、肝糖原含量、运动后血清尿素水平、血乳酸曲线下面积等指标，以测定其抗疲劳作用；每天以上述剂量给小鼠灌胃，同时设立一个辐射对照组，连续灌胃21d后，不同指标选择不同的照射剂量，各组均以同一剂量7射线全身照射一次，照射后继续给予样品。照射前、后第3d和第14d进行外周血白细胞计数，辐照后第3d进行骨髓有核细胞计数和骨髓细胞微核实验，第7d测定血清超氧化物歧化酶（SOD）活力，第14d测定血清溶血素，以测定其抗辐射作用。结果0.36和1．08g／kg·bw剂量能延长小鼠的负重游泳时间，1．08g／kg·bw剂量能显著降低小鼠运动后血清尿素水平、明显降低小鼠的血乳酸曲线下面积；以3GY剂量照射后第3d和第14d中、高剂量组的外周血白细胞计数明显高于辐射模型对照组；第3d，高剂量组小鼠的骨髓有核细胞数明显高于对照组，骨髓细胞微核率明显低于对照组；以6GY剂量照射后第7d，高剞量组小鼠的血清超氧化物歧化酶（SOD）活性明显高于对照组。差异有统计学意义（P〈0．05或P〈0．01）。结论以内苁蓉、刺五加、大枣为主要原料的软胶囊具有抗疲劳、抗辐射作用。