Evaluation of trophoblast HLA-G antigen with a specific monoclonal antibody

A monoclonal antibody to HLA-G has been generated by immunizing HLA-A2.1/human β²-microglobulin (β²m) double transgenic mice with murine L cells transfected with both human β²m and HLA-G. This monoclonal antibody, designated as G233, has been found not to cross-react with other HLA class I antigens when tested on numerous cell lines by flow cytometry. With immunohistology, all populations of extravillous trophoblast (cell columns, interstitial trophoblast, endovascular trophoblast, placental bed giant cells) were stained. An extensive range of adult and fetal tissues was also tested but none reacted with monoclonal antibody G233, including those previously reported to express HLA-G mRNA, indicating that the protein has a highly restricted distribution. Failure to detect HLA-G in the fetal thymus raises the question as to how T-cell tolerance to this antigen is induced. Immunoprecipitation of trophoblast surface proteins with monoclonal antibody G233 revealed a heavy chain of 39 kDa and a light chain of 12 kDa, indicating that HLA-G expressed on the surface of trophoblast is complexed with p2m. However, sequential immunoprecipitation with monoclonal antibody W6/32 followed by monoclonal antibody G233 continued to detect a residual band of 39 kDa, suggesting that trophoblast surface HLA-G may also occur as free heavy chains not associated with p2m. Immunoprecipitation followed by two dimensional gel electrophoresis showed that monoclonal antibody G233 recognizes several iso-forms of HLA-G from trophoblast similar to the characteristic spot array previously described for HLA-G. This monoclonal antibody G233 will be highly useful in future experiments to elucidate the function of HLA-G.     

Differences in leisure-time physical activity levels between blacks and whites in population-based samples: The Minnesota heart survey

Energy expenditure in leisure-time physical activity (LTPA) was measured using the Minnesota LTPA Questionnaire in 35- to 74-year-old black and white residents of Minneapolis-St. Paul, Minnesota. Estimates of the geometric mean LTPA energy expenditure were 129 and 204 kcal per day for black and white men (p<.05) and 91 and 123 kcal per day for black and white women (p<.05). The percentage of individuals expending 2000 kcal or more per week in LTPA was significantly lower in black men than white men (25 vs. 35%; p=.01) but was not different in black versus white women (18 vs. 17%). Although black men and women reported greater occupational physical activity than their white counterparts, LTPA and job activity were unrelated in all race and sex groups. In both races, LTPA energy expenditure declined with age. LTPA increased with level of formal education, and the largest LTPA difference between blacks and whites was observed in those who had a high-school diploma or less. Blacks had lower participation rates than whites in most of the individually assessed physical activities. Additional research is needed on the determinants and promoters of LTPA in population subgroups.The research was funded by National Heart, Lung, and Blood Institute Research Grant R01 HL 23727 and National Research Award T32 HL 07328-10.     

Thoraskopische Koronarchirurgie

Thorascopic coronary surgery has achieved great progress in the past few years. After computer-directed instrument systems were introduced, which enabled high precision and expanded mobility in close quarters, the limitations of conventional endoscopic instruments were largely overcome. Employing modern surgical telemanipulators and endoscopic stabilizers, entirely endoscopic myocardial revascularization of the anterior wall is being successfully performed during off-pump surgery. Longer operating times and conversion rates in about one-fifth of the patients due to technical and anatomical limitations still restrict the use of these new surgical techniques compared to conventional surgery. Development of further technologies and integration of video-assisted navigation systems will broaden the application of computer-assisted instrument systems in the near future.     

In-vitro fertilization and the ovarian hyperstimulation syndrome.

Eight patients who developed severe ovarian hyperstimulation syndrome (OHSS) were identified among 1302 patients undergoing in-vitro fertilization (IVF) over a 1 year period (prevalence of 0.6%); 63% had ultrasonically diagnosed polycystic ovaries (PCO) and 75% were undergoing their first attempt at IVF. Pretreatment with a superactive luteinizing hormone-releasing hormone (LHRH) analogue significantly increased the prevalence of severe OHSS (1.1% versus 0.2%, P less than 0.05) compared with ovarian stimulation with clomiphene citrate and human menopausal gonadotrophin (HMG). The mean serum oestradiol concentration on the day of human chorionic gonadotrophin (HCG) administration was 8200 +/- 2300 pmol/l. A mean of 19.6 +/- 6.8 follicles had been aspirated and 13.1 +/- 7.7 oocytes recovered at transvaginal ultrasound-directed oocyte recovery. All patients had an embryo transfer and luteal support in the form of HCG. The clinical pregnancy rate was 88%, multiple pregnancy rate 71% and implantation rate 63.5 +/- 41.3%. In a group of seven patients who were hospitalized for moderate OHSS during the same period, peak oestradiol levels were significantly lower than in those with severe OHSS (P less than 0.05). Of the group with moderate OHSS, 57% had PCO, the clinical pregnancy rate was 100% and multiple pregnancy rate 43%. Patients with ultrasound-diagnosed PCO have an increased risk of developing OHSS and the dose of HMG administered to them should be minimized. In patients at risk of developing OHSS, progesterone instead of HCG should be used for luteal support. Transfer of a maximum of two embryos or freezing all embryos for transfer in a subsequent cycle may reduce the likelihood of multiple pregnancy.     

Community-wide prevention of cardiovascular disease: education strategies of the Minnesota Heart Health Program

The Minnesota Heart Health Program (MHHP) is a research and demonstration project of population-wide primary prevention of cardiovascular disease. Study goals are to achieve reductions in cardiovascular disease risk factors and morbidity and mortality in three education communities compared with three reference communities. The program in the first of the three intervention communities, Mankato, has been operating for 3 of the planned 5 years. Early objectives of the program have been achieved based on data obtained from population-based random samples surveyed in education and comparison communities. After 2 years of participation, Mankato was significantly more exposed to activities promoting cardiovascular disease prevention. In this town of 38,000 inhabitants, 190 community leaders were directly involved as program volunteers, 14,103 residents (over 60% of adults) attended a screening education center, 2,094 attended MHHP health education classes, 42 of 65 physicians and 728 other health professionals participated in continuing education programs offered by MHHP, and distribution of printed media averaged 12.2 pieces per household. These combined educational strategies have resulted in widespread awareness of MHHP and participation by the majority of the Mankato adult population in its education activities.     

FM sonography in diffuse liver disease: prospective assessment and blinded analysis

FM sonography - a signal-processing technique that uses frequency and phase information as well as amplitude data - shows promise in evaluation of patients with diffuse liver disease. In a prospective blinded review of 37 patients with biopsy-proved liver disease and 42 healthy volunteers, FM sonography was clearly superior to traditional amplitude-based (AM) sonography in distinguishing healthy from diseased subjects. Statistically significant differences were seen in accuracy (FM, 98.7%; AM, 84.8%), sensitivity (FM, 97.3%; AM, 70.3%), and negative predictive value (FM, 97.7%; AM, 78.8%). Our data also suggest that current FM sonographic techniques cannot differentiate among histologic findings associated with different hepatic parenchymal abnormalities. It is unclear, therefore, whether FM imaging can reduce the numbers of patients who require biopsy for diagnosis or the frequency of biopsy procedures in patients with known disease.     

Color vision polymorphism and its photopigment basis in a callitrichid monkey (Saguinus fuscicollis)

The color vision of five saddle-backed tamarins (Saguinus fuscicollis) was studied. Behavioral tests of color discrimination and spectral sensitivity indicate that this species has a color vision polymorphism. Individual monkeys have either dichromatic or trichromatic color vision. Measurements of the spectral sensitivity of cones on this species were made on nine animals with the technique of electroretinogram (ERG) flicker photometry. Both the electrophysiological and the behavioral results suggest that there are four classes of cone pigment in this species. In addition to a short wavelength sensitive cone, apparently common to all tamarins, there are three classes of middle to long wavelength cone (lambda max = 545, 557 and 562 nm). Individual animals have either one or two of the latter types. The color vision variation in this species differs for males and females.     

Entrapment of platelets in the penis during and after erection

Because of the development of hypercoagulability and the deposition of fibrin in the penis during erection a study of the possible role of platelets in this process was undertaken. Platelet response was studied in 9 adult chacma baboons (Papio ursinus) using autologous in vitro indium-111-labelled platelets and sequential scintigraphy of the penis during erection. The blood pooling pattern was obtained using in vivo technetium-99m-labelled red cells in a similar investigation. A statistically significant retention of platelets occurred during and after erection, which could not be attributed to blood pooling (P less than 0,05). Entrapment of platelets could lead to enhanced activity and activation, and might play a significant role in hypercoagulability and fibrin deposition during erection. Therefore platelets could be an important factor in the pathogenesis of ageing impotence.     

The clinical utility of provocative radionuclide oesophageal transit in the evaluation of non-cardiac chest pain.

We combined edrophonium provocative testing with the technique of radionuclide oesophageal transit (RET) in 30 consecutive patients with non-cardiac chest pain (NCCP) and 12 controls. The oesophageal transit time of aqueous technetium-99m sulfur colloid was determined before and after intravenous infusion of 80 micrograms/kg edrophonium chloride (ED). Patient symptoms during provocative RET (P-RET) were recorded. Thirteen (43%) of the patients had abnormal study results, whereas all control subjects had normal results. Three groups considered abnormal were observed: (a) in two patients (6%), the pain was reproduced and transit pre- and post-ED administration was prolonged (greater than 15 s); (b) in six patients (20%), the pain was reproduced, but transit was normal pre- and post-ED; (c) in five patients (17%), transit pre- and post-ED was prolonged, but no pain was reproduced. In five patients (17%), ED prolonged the transit time greater than 15 s without pain, but the baseline transit was normal. Transit time was measurable in 23 patients. Mean pre-ED transit time was 10.2 +/- 7.4 s (mean +/- SD) and post-ED, 12.4 +/- 8.0 s (P = 0.3). We conclude that ED has no significant effect on transit time, and the pain induced by ED rarely correlates with an abnormal transit; P-RET provides additional information to baseline RET, increasing sensitivity, and may be a useful screening method in the evaluation of patients with NCCP.     

Quantitative effects of cerebral infarction on spatial learning in rats.

Outcome following stroke is difficult to measure because the behavioral response to infarction is variable. We hypothesized that cognitive function, such as spatial learning, may be a reproducible and sensitive outcome variable. We developed an animal model of multifocal cerebral ischemia in order to study the effects of infarction on learning. To cause ischemia, several hundred microspheres were injected into the internal carotid arteries of rats. After ischemia, behavior was measured using a global rating and a Morris water maze. Postmortem serial brain sections were stained and the size of the infarctions was measured. We found that intracerebral microspheres caused cortical infarction and an impairment of spatial learning. This impairment was not due to occlusion of the internal carotid artery and was not found in animals who received a sham injection of saline. The degree of learning impairment was not correlated with the volume density of the infarctions or with the volume density of the remaining cerebral hemisphere. The learning impairment clearly differentiated normal from lesioned animals, and the impairment was probably due to a delay in acquisition of spatial information rather than a defect in retention or retrieval. Measurement of learning deficit after cerebral ischemia is an efficient and sensitive method for evaluating new stroke treatments and possibly for exploring structure function relationships.