Interaction of nanobacteria with cultured mammalian cells

Nanobacteria were recently isolated from human blood and commercial fetal bovine serum (FBS) and were located in the -2 subgroup of proteobacteria based upon their 16S rRNA gene sequence. They can be cultured even in the absence of mammalian cells, and have extraordinary properties, like very slow growth rate and an impermeable cell wall, making their detection difficult by standard microbiological techniques. Since they are present in FBS, and thus in cell cultures, it is essential to clarify their effects on cultured mammalian cells. In this study, we show that four out of six nanobacterial isolates from different sera exerted a cytotoxic effect on 3T6 fibroblasts verified by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] viability assay, lactate dehygrogenase (LDH) release and by direct microscopy. The cytotoxic effect of nanobacteria was attenuated after they had been subcultured several times. The cytotoxic effect was similar with all tested murine and human fibroblastoid cell lines. Differential interference contrast and electron microscopy, and FITC staining with specific monoclonal antibodies indicated selective, possibly receptor-mediated adherence, followed by internalization and cytotoxicity in the 3T6 fibroblasts used as a model in these interaction studies. Thus, nanobacteria have a special way of invading mammalian cells: they trigger cells that are not normally phagocytic to engulf them. These organisms seem to be an important cause for cell vacuolization, poor thriving and unexpected cell lysis, problems frequently encountered in mammalian cell culture.     

Effects of cilostazol on lipid and fatty acid metabolism

Abstract Cilostazol is a selective inhibitor of phosphodiesterase III with anti-platelet-aggregatory and vasodilating properties. Randomised, double-blind, placebo-controlled trials in 2702 patients with intermittent claudication demonstrated that cilostazol significantly increased walking distances compared with placebo. Furthermore, the agent has beneficial effects on the serum lipid profile and fatty acid composition in plasma. Consequently, cilostazol may be useful to prevent atherosclerosis from progressing by ameliorating lipid and fatty acid metabolism.     

脂质、血红蛋白、胆红素标本对乙肝病毒DNA荧光定量测定的影响

目的:探讨脂血、高胆红素和溶血标本对乙肝病毒DNA(HBVDNA)荧光定量测定结果的影响。方法:将乙肝大三阳高脂血和非脂血、溶血血清和未溶血血清同时作HBVDNA荧光定量检测;将HBVDNA阴性黄疸血清和HBVDNA阴性正常血清与来自同一份乙肝大三阳血清混合,在相同条件下进行HBVDNA荧光定量。结果:乙肝大三阳溶血与未溶血样本HBVDNA含量都在同一数量级。乙肝大三阳高脂血的HBVDNA含量明显低于对照标本。高黄疸血清、正常对照血清与相同的HBVDNA阳性模板组合后所测得的HBVDNA结果无差异。结论:脂血对HBVD-NA定量测定有严重干扰;溶血样本、高胆红素样本对HBVDNA测定结果无影响。     

Serum contains a potent factor that decreases β-adrenergic receptor-stimulated L-type Ca2+ current in cardiac myocytes

L-type Ca²⁺ current (I _Ca) was measured in cultured atrial myocytes from hearts of adult guinea-pigs using whole-cell voltage clamp. Potentiation of I _Ca induced by -adrenergic stimulation (isoprenaline 2· 10^–7 M) could be completely antagonized by diluted sera (1100 v/v). Half-maximal inhibition of -receptorstimulated I _Ca occurred at about 11000. Basal I _Ca was not affected by serum. Atropine in a concentration (10^–6M) that completely antagonized the anti-adrenergic effect of acetylcholine (ACh, 2·10^–6 M) did not interfere with the effect of serum. In cells dialysed with cyclic adenosine monophosphate (cAMP)-containing (10^–4 M) pipette solution, potentiated I _Ca was insensitive to both ACh and serum. Preincubation of the myocytes with pertussis toxin almost completely abolished the anti-adrenergic effects of both ACh and serum. The potency of serum was not reduced by dialysis. It is concluded that serum contains a factor which, like ACh, inhibits -receptor-stimulated adenylyl cyclase via G_iprotein.A preliminary report of this work has appeared in abstract form [11]     

Renal function in relation to three candidate genes in a Chinese population

We recently found in a white population that the genes encoding angiotensin-converting enzyme (ACE, I/D polymorphism), -adducin (Gly460Trp), and aldosterone synthase (–344C/T) jointly influence renal function. We therefore investigated in a Chinese population the associations between the serum concentrations of creatinine and uric acid and these three genetic polymorphisms. We genotyped 471 ethnic Han Chinese subjects from 125 nuclear families recruited in northern China via random population sampling (75%) and at specialized hypertension clinics (25%). We performed population-based and family-based association analyses using generalized estimating equations (GEE) and quantitative transmission disequilibrium test (QTDT), respectively, while controlling for covariables. The participants were 39.7 years old and included 235 women (49.9%). The blood pressure measured at the subjects homes averaged 126/80 mmHg. Mean values were 71 µmol/l for serum creatinine, 111 ml min^–1 1.73 m^–2 for calculated creatinine clearance, and 236 µmol/l for serum uric acid. With adjustment for covariables, GEE analyses of single genes demonstrated that serum uric acid, but not serum creatinine, was positively associated with the ACE D allele. Serum uric acid concentrations were 15.8 µmol/l (95% confidence interval 3.3–28.2) and 25.7 µmol/l (11.1–40.2) higher in DD homozygotes than in ID and II subjects, respectively. Further GEE analyses of the three genes combined showed that the association between serum uric acid and the ACE polymorphism was confined to carriers of the -adducin Gly and/or aldosterone synthase C alleles. Sensitivity analyses in parents and offspring separately as well as QTDT analyses were confirmatory. Among 114 informative offspring carrying the -adducin Gly allele serum uric acid was significantly and positively associated with the transmission of the ACE D allele (=20.7 µmol/l). In conclusion, the present study extends our previous findings on the combined effects of the three candidate genes and supports the concept that these genetic polymorphisms jointly influence renal function.     

动脉粥状硬化与血清中磷脂酰胆碱特异性磷脂酶C活性的相关性研究

炎症在动脉粥状硬化(AS)形成过程中起关键作用,PC-PLC在炎症反应中扮演重要角色。为探讨PC-PLC活性与动脉粥样硬化的关系,本研究建立了动脉粥状硬化家兔模型,并用解毒活血颗粒灌胃,分别获得正常家兔血清、造模家兔血清和药物家兔血清,检测其中PC-PLC的活性。结果表明,造模组中钙离子依赖性和非钙离子依赖性两种PC-PLC活性均明显高于正常组,而加药组中这两种PC-PLC活性受到显著抑制。说明伴随动脉粥样硬化的发生,PC-PLC活性明显升高,而活血解毒颗粒能减轻动脉粥样硬化的程度,同时能显著抑制PC-PLC活性。     

69,XXY伴严重生长迟缓及眼球突出病例的产前诊断

目的分析孕中期血清筛查数据和胎儿彩色多普勒与染色体异常之间的关系。方法对一孕中期血清产前筛查18三体风险1:10,血清Free-hCGβ异常减低(值为1.29ng/ml即0.08MOM),胎儿系统结构检查提示胎儿小于孕周,头腹围比(1.63)大于正常(1.25),上唇连续性中断延至鼻底的孕妇进行羊膜腔穿刺,抽取羊水进行细胞遗传学检查。结果羊水细胞染色体核型:69,XXY。结论中孕期妇女血清Free-hCGβ异常减低和胎儿头胸围发育不同步、胎儿唇腭裂提示胎儿染色体畸变可能,血清和超声联合筛查有助于染色体疾病的检出。     

新生儿血清瘦素水平变化与胰岛素和生长激素关系的研究

目的　探讨新生儿血清瘦素来源及其与胰岛素和生长激素的关系。方法　采用放射免疫法检测 80例新生儿静脉血和脐血瘦素水平 ,根据不同胎龄新生儿出生体重值分为大于胎龄儿 (LGA)组 2 8例 ,适于胎龄儿 (AGA)组 36例 ,小于胎龄儿 (SGA)组 16例 ;采用Rohrer′s指数 =出生体重 (g)× 10 0 /身长 (cm) 3 估测新生儿营养状态。结果　早产儿血清瘦素水平明显低于足月儿 (0 6 6± 1 0 3ng/mlvs 3 5 9± 2 16ng/ml,P <0 0 1) ;AGA儿血清瘦素水平 (3 0 6± 0 96ng/m1)明显低于LGA儿(4 0 3± 2 2 2ng/m1) ,而高于SGA儿 (1 13± 1 98ng/m1) ;足月新生儿血清瘦素水平与Rohrer′s指数、新生儿体重、胎龄 ,血清胰岛素、生长激素水平呈显著正相关。结论　新生儿体内瘦素主要来源于自身脂肪组织 ,它反映新生儿的生长营养状态 ,推测瘦素可能通过胰岛素、生长激素共同调节新生儿的生长发育 ,在胎儿和新生儿生长发育中起重要作用。     

Screening 34 Peanut Introductions for Allergen Content Using ELISA

Peanut is one of the most allergenic foods and reports of accidental ingestion of peanuts in unsuspected food are increasing. No information is available on the allergen content of peanut germplasm grown commercially and used in the food and confectionery industry. The objectives of this study were: (1) to determine the allergen contents of 34 peanut introductions (PI); and (2) to identify naturally occurring allergen-free and/or low or hypoallergenic peanuts germplasm. A basic ELISA protocol was utilized to detect the presence of antigens in the peanut lines using a pool of human sera from patients with documented history of peanut allergy. Two naturally occurring low, or hypo-allergenic germplasm were identified as PI 261942 and PI 338386. Both are Valencia market types with total allergen content significantly lower (P ≤0.05) than that of PI 119880 (0.550), PI 119876 (0.415) and PI 118991 (0.410) three Valencia market types and PI 262111 (0.485), a Virginia market type. No allergen-free PI was found. Allergen content of peanut lines from Bolivia and Paraguay were significantly (P ≤0.05) different to those from Venezuela. No significant difference was observed in the allergen content of the four market types.     

Serum protein concentration and oncotic pressure relationship in the rat

Summary A formula relating oncotic pressure torat serum protein concentration was derived and found to be in complete agreement with the Landis-Pappenheimer formula derived from measurements onhuman plasma.This study was supported by the Danish Medical Research Council and Johan and Hanne Weimann's legacy.