Isolated human hepatocytes in culture display markedly different gene expression patterns depending on attachment status

In vitro human hepatocyte cultures are a key tool in the investigation of xenobiotic toxicity and metabolism. In most in vitro hepatocyte studies, the cells are allowed to adhere to an extracellular matrix, such as collagen. Unfortunately, the ability of freshly isolated hepatocytes to adhere to collagen varies from donor to donor. We used microarray analysis to determine what gene expression differences exist between hepatocytes in suspension and hepatocytes attached to collagen. Results from different donors showed a considerable difference in gene expression patterns between the two hepatocyte populations. In addition, we also compared the gene expression profiles of hepatocytes in culture with liver tissue. The results showed that both hepatocytes in suspension and hepatocytes attached to collagen display significant gene expression differences compared with liver tissue. Finally, we show that both populations of hepatocytes are responsive to dexamethasone and regulate some of the same genes. Overall, our results suggest that either significant gene expression changes occur in isolated hepatocytes or that suspended and attached cells represent different populations of hepatocytes found in intact livers.     

Promotion of mouthguards among amateur football players in Victoria

Abstract: Wearing mouthguards while playing contact sports reduces the prevalence and severity of orofacial injuries. We report on a 1991–92 promotion of mouthguard use among amateur football players in Melbourne. Four questionnaires were completed by players (1991 season: middle: n =638, 49 per cent response; late: n =770, 51 per cent; 1992: early: n =505, 26 per cent; late: n =698, 54 per cent), in four age groups (16–19, 20–23, 24–29, 30–44 years), about perceived mouthguard value and usage, injury experience, and awareness of a promotional message. Over the two seasons, mouthguard ownership and consistent wear increased and dental injuries decreased. Ownership of mouthguards increased among: 20–23 year olds and 30–44 year olds (13 and 16 per cent); managers, tradespersons, salespersons, labourers and the unemployed (14–19 per cent); residents of northern and western regions (12 and 13 per cent); those with 6 to 10 years of education (29 per cent); players in old boys' networks (13 per cent); and those with English as a first language (8 per cent). Consistent wear was high at matches (89 per cent) but low at training (13 per cent), owing to perceived lack of physical contact. Of dental injuries reported by 25–31 per cent, fractures predominated over luxations and avulsions. Fractures and avulsions were more common at matches than training; luxations predominated at training. When a mouthguard was not worn, the likelihood of a fractured or avulsed tooth was at least twice that when a mouthguard was worn. Some injuries occurred despite the wearing of mouthguards. The promotional message appeared effective in increasing mouthguard use; newspapers and football journals provided effective message locations for players, and ground signs and the electronic scoreboard for spectators.     

Diagnostic, prognostic and therapeutic value of Doppler echocardiography in pulmonary embolism. Apropos of 41 cases

In 41 patients with acute or recent pulmonary embolus (PE), a prospective study was conducted with cardiac Doppler sonography (CDS). The test is performed first in all patients, then immediately following a treatment fibrinolytics in 10 of them. The PE is confirmed by pulmonary angiography in 39 patients and embolectomy for the 2 patients who presented a mobile thrombus in the right atrium. A tricuspid insufficiency (TI) is almost always recorded by continuous Doppler: the systolic pulmonary artery pressure (sPAP) is calculated from the maximum velocity of the regurgitating tricuspid flow, using Bernouilli's equation. The correlation between Doppler sPAP and sPAP obtained by catheterization is: r = 0.95 (n = 34; p less than 0.001). A severe pulmonary hypertension, corresponding to a TI exceeding 3.5 m/s is related to a pulmonary migration occurring on a pathological heart. There is a satisfactory relationship between the elevation of the Doppler sPAP and the degree of pulmonary obstruction in the "Pe on healthy heart" population: r = 0.6 (n = 29; p less than 0.001). The dilated right ventricle observed on the sonogram (RV) is proportional to the severity of the angiographic image: correlation between the RV/LV ratio and the percentage of vascular obstruction is r = 0.73 (n = 27; p less than 0.001). The drop in the sPAP and the regression of the symptoms of pulmonary heart noted on CDS after fibrinolysis, correspond to a significant improvement of the pulmonary perfusion. The CDS seems to be absolutely necessary in PE because of the major and reliable informations it provides.     

Image-directed percutaneous biopsies with a biopsy gun

Core tissue for histologic study is believed by many pathologists to be more diagnostic than material from needle aspiration. Recently, a biopsy "gun" has been introduced, which simplifies core biopsies. With this device, 182 biopsies of multiple anatomic sites were performed with ultrasonic, computed tomographic, and fluoroscopic guidance and 18-gauge needles. High-quality histopathologic specimens were obtained in 177 of the biopsies, and diagnostic target tissue was obtained in 167. Only three significant complications occurred: one bleeding complication that required transfusion and two cases of pneumothorax that necessitated placement of chest tubes. The biopsy gun eliminated the disjointed movements of conventional "skinny" needle biopsies, and none of the samples demonstrated significant "crush" artifact or obscuring blood, problems that are commonly associated with manual biopsy techniques. Patient discomfort was decreased with this system compared with that of manual biopsies, and the total procedure time was reduced. Because of these distinct advantages, the authors now use the biopsy gun exclusively for all percutaneous biopsies and recommend that other institutions consider the use of this biopsy method.     

新生期大白鼠皮下注射谷氨酸单钠对成年后下丘脑α-促黑素细胞激素神经元免疫反应性的影响

本文用免疫组化方法研究了新生期大白鼠注射谷氨酸单钠(MSG)对成年后下丘脑α-促黑素细胞激素(α-MSH)免疫反应神经元的影响,结果显示MSG处理以后下丘脑弓状核区α-MSII免疫反应神经元减少甚至完全消失,但不影响下丘脑背外侧区的α-MSH神经元群。文中还讨论了这两群α-MSH神经元的生理作用。     

Linkage of the MHC to familial multiple sclerosis suggests genetic heterogeneity. The Multiple Sclerosis Genetics Group

Multiple sclerosis (MS) is a demyelinating autoimmune disease of the central nervous system. While its etiology is not well understood, genetic factors are clearly involved. Until recently, most genetic studies in MS have been association studies using the case-control design testing specific candidate genes and studying only sporadic cases. The only consistently replicated finding has been an association with the HLA-DR2 allele within the major histocompatibility complex (MHC) on chromosome 6. Using the genetic linkage design, however, evidence for and against linkage of the MHC to MS has been found, fostering suggestions that sporadic and familial MS have different etiologies. Most recently, two of four genomic screens demonstrated linkage to the MHC, although specific allelic associations were not tested. Here, a dataset of 98 multiplex families was studied to test for an association to the HLA-DR2 allele in familial MS and to determine if genetic linkage to the MHC was due solely to such an association. Three highly polymorphic markers (HLA-DR, D6S273 and TNFbeta) in the MHC demonstrated strong genetic linkage (parametric lod scores of 4.60, 2.20 and 1.24, respectively) and a specific association with the HLA-DR2 allele was confirmed (TDT; P < 0.001). Stratifying the results by HLA-DR2 status showed that the linkage results were limited to families segregating HLA-DR2 alleles. These results demonstrate that genetic linkage to the MHC can be explained by the HLA-DR2 allelic association. They also indicate that sporadic and familial MS share a common genetic susceptibility. In addition, preliminary calculations suggest that the MHC explains between 17 and 62% of the genetic etiology of MS. This heterogeneity is also supported by the minority of families showing no linkage or association with loci within the MHC.      

CTLA-4 is required for the induction of high dose oral tolerance

Mucosal and systemic administrations of high dose antigens induce long- lasting peripheral T cell tolerance. We and others have shown that high dose peripheral T cell tolerance is mediated by anergy or deletion and is preceded by T cell activation. Co-stimulatory molecules B7-1 (CD80)/B7-2 (CD86) and their counter-receptors CD28/CTLA-4 play pivotal roles in T cell activation and immune regulation. In the present study, we examined the roles of the B7 co-stimulation pathway in the generation of high dose peripheral T cell tolerance. We found that blocking B7:CD28/CTLA-4 interaction at the time of tolerance induction partially prevented T cell tolerance, whereas selective blockade of B7:CTLA-4 interaction completely abrogated peripheral T cell tolerance induced by either oral or i.p. antigens. These results suggest that CTLA-4-mediated feedback regulation plays a crucial role in the induction of high dose peripheral T cell tolerance.      

Screening for proteins with polyglutamine expansions in autosomal dominant cerebellar ataxias

Expansion of trinucleotide CAG repeats coding for polyglutamine has been implicated in five neurodegenerative disorders, including spinocerebellar ataxia (SCA) 1 and SCA3 or Machado-Joseph disease (SCA3/MJD), two forms of type I autosomal dominant cerebellar ataxias (ADCA). Using the 1C2 antibody which specifically recognizes large polyglutamine tracts, particularly those that are expanded, we recently reported the detection of proteins with pathological glutamine expansions in lymphoblasts from another form of ADCA type I, SCA2, as well as from patients presenting with the distinct phenotype of ADCA type II. We now have screened a large series of patients with ADCA or isolated cases with cerebellar ataxia, for the presence of proteins with polyglutamine expansions. A 150 kDa SCA2 protein was detected in 16 out of 40 families with ADCA type I. This corresponds to 24% of all ADCA type I families, which is much more frequent than SCA1 in this series of patients (13%). The signal intensity of the SCA2 protein was negatively correlated to age at onset, as expected for an expanded and unstable trinucleotide repeat mutation. The disease segregated with markers closely linked to the SCA2 locus in all identified SCA2 families. In addition, a specific 130 kDa protein, which segregated with the disease, was detected in lymphoblasts of patients from nine families with ADCA type II. It was also visualized in the cerebral cortex of one of the patients, demonstrating its translation in the nervous system. Finally, no new disease-related proteins containing expanded polyglutamine tracts could be detected in lymphoblasts from the remaining patients with ADCA or isolated cases with cerebellar ataxia.      

Improvement in ischemic myocardial blood flow following a new calcium antagonist

The effect of FR 7534, a new dihydropyridine CA2+ antagonist, nitroglycerin, and dipyridamole have been compared on coronary collateral function in pentobarbital-anesthetized open-chest dogs following acute ligation of the left anterior descending coronary artery. Measurements of tissue blood flow using radioactive microspheres were made 60 min postligation after low and high doses of each drug and at the higher dose with methoxamine added to to return mean aortic pressure. FR 7534 treatment increased subepicardial, subendocardial, and transmural tissue flow by 100% in the central ischemic zone when perfusion pressure was controlled by methoxamine. Nitroglycerin also increased ischemic tissue flow, but to a lesser degree, 18% transmurally. Dipyridamole produced no significant change. Tissue flow in normal myocardium was similarly increased by FR 7534 and dipyridamole but slightly reduced by nitroglycerin. FR 7534 and nitroglycerin also increased retrograde coronary pressure when aortic presure was maintained constant. FR 7534, but not nitroglycerin, increased ischemic contractile force. In this model, FR 7534 may produce greater effects than nitroglycerin in increasing blood supply to ischemic myocardium delivered by endogenous collaterals especially when aortic perfusion pressure was controlled.