Criteria for Assessment of Functional Impairment in Patients with Cirrhosis of the Liver*

Abstract. Fourty-seven patients with cirrhosis of the liver and 58 normal subjects were studied in order to find quantitative criteria to assess the severity of the functional impairment in this disease. The special investigations included hepatic haemodynamics, partial functions and liver volume, which was measured in frontal and right lateral scans. – In contrast to normal subjects liver volumes in cirrhosis were not correlated significantly with the body weight (r=–0.05), with the galactose elimination capacity (Tygstrup's method, r= 0.15) or with the sulfobromophthalein transport maximum (Wheeler's method, r=–0.40). Large livers, however, were associated with corresponding increases in splanchnic oxygen uptake (r= 0.74, P < 0.001), even though hepatic blood flow estimated with the indocyanine green infusion and extraction technique appeared to vary independently (r= 0.40, P > 0.05). – It may be inferred from these data that patients with cirrhosis can be further characterized on the basis of liver volume. Compared to small livers hepatomegaly shows a dissociation between oxygen uptake and functional capacity. Thus, with existing methods the severity of functional impairment in cirrhosis may be quantitatively investigated. Such information should be relevant for prognostic stratification and endpoint definition of therapeutic trials.     

Xenotransplantation literature update June-October 2010

Schneider MKJ, Seebach JD. Xenotransplantation literature update June – October 2010. Xenotransplantation 2010; 17: 481–488. © 2010 John Wiley & Sons A/S.     

A recombinant probiotic for treatment and prevention of cholera

BACKGROUND & AIMS: We have developed a therapeutic strategy based on molecular mimicry of host receptors for bacterial toxins on the surface of harmless gut bacteria. In the present study, this has been applied to the development of a recombinant probiotic for treatment and prevention of cholera, caused by Vibrio cholerae. METHODS: We expressed glycosyltransferase genes from Neisseria gonorrhoeae and Campylobacter jejuni in a harmless Escherichia coli strain, resulting in production of a chimeric lipopolysaccharide terminating in a mimic of the ganglioside GM(1). RESULTS: The recombinant bacterium was capable of binding cholera toxin, a sine qua non of virulence, with high avidity; when tested with purified cholera toxin, it was capable of adsorbing >5% of its own weight of toxin in vitro. Administration of the GM(1)-expressing probiotic also protected infant mice against challenge with virulent V cholerae, even when treatment was delayed until after establishment of infection. When treatment commenced 1 hour after challenge, 12 of 12 mice given the probiotic survived, compared with only 1 of 12 for control mice (P < .00001). CONCLUSIONS: Toxin-binding probiotics such as that described here have considerable potential for prophylaxis and treatment of cholera in humans.     

Intestinal absorption in health and disease--sugars

Carbohydrates are mostly digested to glucose, fructose and galactose before absorption by the small intestine. Absorption across the brush border and basolateral membranes of enterocytes is mediated by sodium-dependent and -independent membrane proteins. Glucose and galactose transport across the brush border occurs by a Na(+)/glucose (galactose) co-transporter (SGLT1), whereas passive fructose transport is mediated by a uniporter (GLUT5). The passive exit of all three sugars out of the cell across the basolateral membrane occurs through two uniporters (GLUT2 and GLUT5). Mutations in SGLT1 cause a major defect in glucose and galactose absorption (glucose-galactose Malabsorption), but mutations in GLUT2 do not appear to disrupt glucose and galactose absorption. Studies on GLUT1 null mice and Fanconi-Bickel patients suggest that there is another exit pathway for glucose and galactose that may involve exocytosis. There are no known defects of fructose absorption.     

Rescue of Infectious Sindbis Virus by Yeast Spheroplast-Mammalian Cell Fusion

Sindbis virus (SINV), a positive-sense single stranded RNA virus that causes mild symptoms in humans, is transmitted by mosquito bites. SINV reverse genetics have many implications, not only in understanding alphavirus transmission, replication cycle, and virus-host interactions, but also in biotechnology and biomedical applications. The rescue of SINV infectious particles is usually achieved by transfecting susceptible cells (BHK-21) with SINV-infectious mRNA genomes generated from cDNA constructed via in vitro translation (IVT). That procedure is time consuming, costly, and relies heavily on reagent quality. Here, we constructed a novel infectious SINV cDNA construct that expresses its genomic RNA in yeast cells controlled by galactose induction. Using spheroplasts made from this yeast, we established a robust polyethylene glycol-mediated yeast: BHK-21 fusion protocol to rescue infectious SINV particles. Our approach is timesaving and utilizes common lab reagents for SINV rescue. It could be a useful tool for the rescue of large single strand RNA viruses, such as SARS-CoV-2.     

13C-methacetin and 13C-galactose breath tests can assess restricted liver function even in early stages of primary biliary cirrhosis

Objective. The ¹³C-methacetin breath test quantitatively evaluates cytochrome P450-dependent liver function. The ¹³C-galactose breath test non-invasively measures the galactose oxidation capacity of the liver. The aim of this study was to find out whether these breath tests are sensitive parameters also in non-cirrhotic patients with primary biliary cirrhosis.

Material and methods. Nineteen patients with early-stage primary biliary cirrhosis (no cirrhotic alterations in the liver biopsy, Ludwig stage I–III) and 20 healthy controls underwent the ¹³C-methacetin and ¹³C-galactose breath tests.

Results. Patients with primary biliary cirrhosis metabolized less ¹³C-methacetin than controls (cumulative recovery within 30 min 7.5±2.4% versus 14.0±2.6%; p<0.001). When a cut-off?>?9.8% was used for the cumulative recovery after 30 min, the methacetin breath test reached 84.2% sensitivity and 95.0 specificity. In the ¹³C-galactose breath test, the percentage recovery at 60 min in patients was 3.1±1.3%/h, and 6.3±1.1%/h in controls (p<0.001). Using a cut-off?>?4.7%/h, the galactose breath test reached 89.5% sensitivity and 95.0 specificity.

Conclusions. In non-cirrhotic, early-stage, primary biliary cirrhosis the ¹³C-methacetin breath test and the ¹³C-galactose breath test reliably indicate decreased liver function. The ¹³C-galactose breath test can also predict the histological score.     

移植培养人肝细胞对急性肝衰竭小鼠的保护作用

目的探讨腹腔移植微载体粘附培养人肝细胞对Ｄ氨基半乳糖（ＤＧａｌ）诱导的急性肝衰竭小鼠的保护作用．方法采用胶原酶灌流法分离人肝细胞，胶原被覆的微载体Ｃｙｔｏｄｅｘ３加以培养，经腹腔注射２×１０６个肝细胞及载体，观察急性肝衰竭小鼠（ｎ＝３２，２５ｇ／ｋｇＤＧａｌ诱导４ｈ）７２ｈ存活率、血清ＡＬＴ、总胆红素以及肝脏病理变化．结果与只接受微载体而无粘附肝细胞的对照组比较，腹腔移植培养肝细胞对肝衰竭小鼠的存活率有明显改善（６５％ｖｓ０％），其血清ＡＬＴ（ＩＵ／Ｌ，２１７４４±２６３０ｖｓ４２６３１±４９２８，Ｐ＜００５）及总胆红素（ｍｍｏｌ／Ｌ，２６９１±３７６ｖｓ４１６８±３８３，Ｐ＜００５）较低．肝组织病理改变较轻．结论腹腔移植微载体粘附培养肝细胞可提供代谢支持，使ＤＧａｌ损伤的小鼠肝功能恢复．     

实验性糖性与萘性白内障动物模型的制作及评价

目的:选择适当的动物、试剂及给药途径,制备能动态显示白内障形成的动物模型。方法:（1）豚鼠,0.4％d-半乳糖,0.2mL/（眼·d）,球后注射19天;（2）大鼠,50％　d-半乳糖腹腔注射,25g/（kg·d）,共24天;（3）家兔,30％萘混悬液灌胃,2.65mL/（kg·d）,共8天。应用裂隙灯显微镜观察不同动物模型晶状体混浊度变化并测定其生化指标。结果:成功获得了糖性及萘性白内障动物模型。结论:此两类模型是深入研究白内障的发病机理及药效学的可靠实验手段。     

利用兔模型研究糖基化对冷藏保存人血小板体内生存的影响

利用兔动物模型研究了糖基化对4℃冷藏保存人血小板体内生存的影响。静脉输注棕榈酸乙酯(0.75g/kg)抑制兔网状内皮系统以建立兔动物模型;应用核素标记法检测血小板生存时间;取浓缩人血小板悬液(2×1012/L),添加5g/L的尿嘧啶二磷酸半乳糖(UDGGal),置4℃冰箱冷藏保存,尔后测定输入兔模型体内的冷藏人血小板的生存率。结果表明:添加UDGGal的人血小板,冷藏保存10天后输入兔模型体内的生存时间显著高于空白对照组(P<0.01),而与新鲜血小板对照组相近(P>0.05);输注兔模型体内2小时时的血小板生存率在新鲜血小板对照组、冷藏糖基化血小板组和冷藏空白对照组分别为(68.9±8.5)%、(65.4±8.0)%和(5.0±2.6)%。结论:尿嘧啶二磷酸半乳糖能保护冷藏保存的人血小板,延长冷藏人血小板在兔模型体内的生存时间。