Allosteric interactions between the membrane-bound acetylcholine receptor and chemical mediators: equilibrium measurements

An approach to equilibrium dialysis measurements has been developed which enables one to study the interaction of chemical mediators with the membrane-bound acetylcholine receptor and to gain information of a type previously obtainable only with soluble proteins. Equilibrium dialysis experiments conducted at pH 7.0,4 degrees C, and mu = 0.18 M, with electroplax membrane preparations from Electrophorus electricus revealed apparently homogeneous binding isotherms for decamethonium with dissociation constants in the range of 0.2-0.4 muM. The following new information has been obtained. (1) The activators of neural transmission, decamethonium and carbamylcholine, occupy overlapping binding sites. (2) These activators and the inhibitors, alpha-bungarotoxin and d-tubocurarine, compete for only one-half of the sites available to them even through the stoichiometry of these is 1:1 as measured with decamethonium (a reversibly binding activator) and alpha-bungarotoxin (an irreversible specific inhibitor). Different receptor molecules, preexisting nonequivalent binding sites, or an allosteric mechanism involving ligand-induced conformational changes are often considered to account for such observations.     

Early repair of diaphragmatic hernia to correct associated anomalies

The case of a seven-month-old infant is reported who had an eventration of the left hemidiaphragm associated with two bronchopulmonary foregut anomalies, duplication of the esophagus, and intralobar sequestration of the left upper lobe. The literature indicates that eventration of the diaphragm is associated with pulmonary sequestration in up to 50% of patients. Since the eventration itself obscures roentgenographic evidence of other foregut anomalies, the concomitant diagnosis and treatment of possible foregut anomalies is another justification for surgical repair of eventration of the diaphragm.     

Autoantibodies to the insulin receptor. Effect on the insulin-receptor interaction in IM-9 lymphocytes

The serum of some patients with insulin-resistant "diabetes" contains antibodies that bind to and block the cell membrane receptors for insulin. In this report, we have characterized the effects of the antireceptor antibodies on the interaction of (125)I-insulin with its receptor on the human lymphoblastoid cell line IM-9. Up to 95% of specific insulin binding can be inhibited by pretreatment of the cells with these immunoglobulins. The onset of the inhibitory effect is time- and temperature-dependent, and the effect is reversed extremely slowly if the cells are suspended in a large excess of antibody-free buffer. These features of antibody binding can be easily distinguished from those for insulin binding to its receptor. The inhibitory effect of the antibodies can be reversed by exposure of the cells to conditions known to elute surface immunoglobulins. The three antireceptor sera studied appear to alter the insulin-receptor interaction in different ways. Two antisera markedly reduce receptor affinity through combined effects on the insulin association and dissociation rates, and, additionally, have smaller effects on available receptor number. A third antiserum primarily affects available receptor number and has little effect on receptor affinity. All three antisera inhibit the capacity of insulin to promote negatively cooperative site-site interactions among insulin receptors. The data suggest that these autoantibodies to the insulin receptor bind to different determinants on the receptor and may therefore be useful as unique probes of insulin receptor structure and function.     

Pancreatic cancer screening. Analysis of the problem and the role of radionuclide imaging

The clinical, surgical, and pathologic findings in a five year prospective study of 192 patients referred with a high probability of pancreatic cancer are reported. We have defined the requirements of any pancreatic imaging procedure as its ability to distinguish a normal pancreas from pancreatic cancer or chronic pancreatitis and the capability of detecting tumors less than 5 cm in diameter. There was a 47 percent incidence of pancreatic disease (27 percent pancreatic cancer and 20 percent chronic pancreatitis). Prospective radionuclide imaging as routinely performed was found to be of little clinical value in this patient population; it was neither specific nor sensitive to pancreatic cancer or chronic pancreatitis. Preliminary data with longitudinal multiplane emission tomography show an improved diagnostic accuracy and the ability to detect resectable tumors, but its efficacy needs to be prospectively compared with other screening tests on a carefully defined patient population.     

Mucinous biliary obstruction associated with a cystic adenocarcinoma of the pancreas

Obstruction of the biliary tree by large amounts of mucinous material is reported in a patient with a mucin-producing cystic adenocarcinoma of the pancreas invading the common bile duct. Although preoperative endoscopic retrograde cholangiography suggested bile duct invasion, mucinous obstruction, as in the previous cases in which it has been reported, was not suspected. Mucinous obstruction, although unusual, should be considered in patients with cystic carcinomas of the pancreas in the presence of jaundice or episodes of cholangitis.     

A comparison of three strains of Holstein-Friesian cows grazed on pasture: growth, development, and puberty

With the introduction of a protein milk payment system in New Zealand in 1988, there was an influx of North American (NA) Holstein-Friesian (HF) genetics into New Zealand (NZ) dairy herds, leading to an increase in the average percentage of NA genetics in NZ HF cows—from 2% in 1980 to 38% in 1999. Of interest has been the effect this change has had on farm profitability and on the management required for these animals, as well as the phenotypic changes that have occurred within the national herd under the breeding programs operated in NZ from 1970 to 1990. The objective of this study was to quantify differences in body dimensions, body weights, and puberty-related parameters among 3 strains of HF, representing animals of NZ origin representative of the genetics present in 1970 and 1990 and of NA origin with 1990s genetics. A total of 172 animals born in 1999 were compared. The strains were 1) NZ70, a strain of NZ Friesian (average 7% NA genetics) equivalent to high-genetic-merit (high Breeding Worth) cows farmed in the 1970s; 2) NZ90, a strain of HF of NZ origin (average 24% NA genetics) typical of the animals present in the 1990s; and 3) NA90, a strain of HF of NA origin (average of 91% NA genetics) typical of animals present in the 1990s. The differences in BW among all strains were significant at 6 and 12 mo of age. At 15 and 24 mo, the 2 NZ strains were significantly lighter than the NA90 animals. At 24 mo of age (i.e., prior to first calving), the NA90 strain animals (BW = 515 kg) were 22 and 34 kg heavier than the NZ90 and NZ70 strains. The body length of the NA90 strain was greater than either of the 2 NZ strains; the differences among the NA90 strain and the 2 NZ strains varied from 2 to 6 cm, with the differences generally being greater at older ages. The trend in heart girth difference among strains was similar to that observed for body length. The wither height of the NA90 animals was greater than that of the NZ strains by 1 to 7 cm, although there was no significant difference between the NA90 and NZ90 strains at birth. At puberty the NA90 heifers were 20 d older and 20 kg heavier than the NZ90 heifers, which in turn were 25 kg and 25 d older than the NZ70 heifers. The NA90 strain had a heavier mature body weight, and their older age at puberty suggested either that they mature later or that, under pastoral conditions, their growth rate is limited by their inability to consume sufficient metabolizable energy as grazed pasture, with a consequent delay in puberty. Results from this study will be useful in revising target BW in growing heifers of different germplasm.     

Assessment of regional left ventricular long-axis motion with MR velocity mapping in healthy subjects

We present a case of trichothiodystrophy associated with a hypereosinophilic syndrome. This case had been followed for 30 years. Trichothiodystrophy was characterized by lamellar ichthyosis, hair and nail dystrophies, kyphoscoliosis, congenital luxation of the hip. The hypereosinophilic syndrome was characterized by an itching urticaria-like eruption. Although the patient's general condition had remaining stable for 30 years, during the last year it worsened and the patient suddenly died. The authors discuss about the significance of this association.     

Enhanced T-cell immunogenicity and protective efficacy of a human immunodeficiency virus type 1 vaccine regimen consisting of consecutive priming with DNA and boosting with recombinant fowlpox virus

The induction of human immunodeficiency virus (HIV)-specific T-cell responses is widely seen as critical to the development of effective immunity to HIV type 1 (HIV-1). Plasmid DNA and recombinant fowlpox virus (rFPV) vaccines are among the most promising safe HIV-1 vaccine candidates. However, the immunity induced by either vaccine alone may be insufficient to provide durable protection against HIV-1 infection. We evaluated a consecutive immunization strategy involving priming with DNA and boosting with rFPV vaccines encoding common HIV-1 antigens. In mice, this approach induced greater HIV-1-specific immunity than either vector alone and protected mice from challenge with a recombinant vaccinia virus expressing HIV-1 antigens. In macaques, a dramatic boosting effect on DNA vaccine-primed HIV-1-specific helper and cytotoxic T-lymphocyte responses, but a decline in HIV-1 antibody titers, was observed following rFPV immunization. The vaccine regimen protected macaques from an intravenous HIV-1 challenge, with the resistance most likely mediated by T-cell responses. These studies suggest a safe strategy for the enhanced generation of T-cell-mediated protective immunity to HIV-1.     

Participation of phosphoinositide-specific phospholipase Cgamma1 and STAT1 protein in the formation of latent complexes of signal proteins

It is known that the growth factor activates appropriate membrane receptors which become starting points of cascades of protein-protein interactions leading to cellular response. Recent data suggest that different signalling pathways may cross-talk during the cellular response. Here we show that phosphoinositide-specific phospholipase C gamma 1, one of the key elements in phosphoinositide pathway of signal transduction, is physically associated with members of the STAT pathway. The precipitation of phospholipase C gamma 1, using polyclonal antibody in A-431 cells, leads to co-immunoprecipitation of STAT1 alpha and STAT1 beta, as well as STAT3. The formation of such complexes was observed in both unstimulated and EGF stimulated cells. The participation of SH3-domains in the formation of such complexes is discussed.