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Taste-sensing type 2 receptors (TAS2Rs) have been implicated in extraoral functions. Airway smooth muscle expresses TAS2Rs and is strongly relaxed by TAS2R agonists. We hypothesised that TAS2R agonists might affect vascular smooth muscle as well. Moreover, the general pharmacological profile of TAS2R agonists, which are used to investigate the functions of TAS2R׳s, are undefined. The aim of this study was to pharmacologically characterise the effects of five prototype TAS2R agonists in vascular smooth muscle. Responses to the TAS2R agonists were investigated in guinea-pig aorta and taenia coli, mouse aorta (wild-type and caveolin-1(-/-) mice) and human pulmonary arteries. Chloroquine, denatonium, dextromethorphan, noscapine and quinine, agonists for TAS2R3, TAS2R4, TAS2R10 and TAS2R14, induced strong endothelium-independent relaxations (responses between 82-96% of maximal relaxations) in phenylephrine pre-contracted guinea-pig aorta that persisted in the presence of L-type Ca(2+) and KCa1.1-channel blockers. Experiments in guinea-pig taenia coli revealed that denatonium and quinine also inhibited relaxations to phenylephrine, indicating antagonism of α-adrenoceptors. Only chloroquine and noscapine mediated relaxations when the guinea pig aorta was pre-contracted by U-46619 or PGF2α. Relaxations to chloroquine and noscapine after U-46619 pre-contractions were however markedly impaired in aortae from caveolin-1(-/-) mice. Chloroquine and noscapine mediated relaxations of human pulmonary arteries that expressed also mRNA for TAS2R3, TAS2R4, TAS2R10 and TAS2R14, at levels similar to that of the α1A adrenoceptor. Notwithstanding whether TAS2Rs are involved or not, TAS2R agonists have profound effects on vascular smooth muscle. Chloroquine and noscapine are of special interest as their effects cannot be accounted for by conventional pathways. (Less)

Atherosclerosis of the thoracic aorta is an independent risk factor for stroke after cardiac surgery. No attention had so far been paid to its topography. The relationship between the topography of aortic atherosclerosis and stroke was studied in patients admitted for coronary surgery.The extent and location of atherosclerosis in the ascending aorta and arch was assessed intraoperatively with epiaortic ultrasound and transesophageal echocardiography in 611 consecutive patients. They were followed for 5.5 +/- 1.7 years (mean +/- SE), amounting to 3,358 patient-years.The incidence of early postoperative (30-day) stroke was 6.4% in patients with atherosclerosis of the ascending aorta and 1.5% in those without (p = 0.004). The five-year stroke-free survival rates (30 days after the operation) for patients without aortic disease, with less than 50%, and with greater than 50% of the ascending aorta affected, were 95.3 +/- 0.9%, 91.8 +/- 2.1%, and 65.0 +/- 14.6%, respectively (p0.0001).Atherosclerosis of the ascending aorta stands out as a predictor of late stroke. High risk is predominantly linked to atheromas in its distal part and lesser curvature.

Background In contrast to the extensive knowledge about human natural killer (NK) cells in peripheral blood, relatively little is known about NK cells in the human lung. Knowledge about the composition, differentiation, and function of human lung NK cells is critical to better understand their role in diseases affecting the lung, including asthma, chronic obstructive pulmonary disease, infections, and cancer. Objective We sought to analyze and compare the phenotypic and functional characteristics of NK cells in the human lung and peripheral blood at the single-cell level. Methods NK cells in human lung tissue and matched peripheral blood from 132 subjects were analyzed by using 16-color flow cytometry and confocal microscopy. Results CD56 dim CD16 + NK cells made up the vast majority of NK cells in human lungs, had a more differentiated phenotype, and more frequently expressed educating killer cell immunoglobulin-like receptors compared with NK cells in peripheral blood. Despite this, human lung NK cells were hyporesponsive toward target cell stimulation, even after priming with IFN-α. Furthermore, we detected a small subset of NK cells expressing CD69, a marker of tissue residency. These CD69 + NK cells in the lung consisted predominantly of immature CD56 bright CD16 − NK cells and less differentiated CD56 dim CD16 + NK cells. Conclusion Here, we characterize the major NK cell populations in the human lung. Our data suggest a model in which the majority of NK cells in the human lung dynamically move between blood and the lung rather than residing in the lung as bona fide tissue-resident CD69 + NK cells.

OBJECTIVESThe aim of the study was to evaluate the risk from calcified atheromas in the ascending aorta, and the extent and topography of the disease in the development of stroke after cardiac surgery.BACKGROUNDPostoperative stroke constitutes a serious problem in cardiac surgery, and atherosclerosis of the ascending aorta is an important risk factor.METHODSBefore surgical manipulation epiaortic echocardiographic ultrasound was performed to evaluate the ascending aorta in 921 consecutive patients undergoing cardiac surgery. The presence of calcification, location of atheroma, extent of the disease and clinical variables including postoperative stroke were recorded prospectively.RESULTSA total of 26.2% of the patients had atherosclerosis of the ascending aorta, and in 44.4% of them more than one of 12 possible segments was involved. Logistic regression showed that atherosclerotic disease in the ascending aorta was the most important predictive factor for postoperative stroke. The incidence of stroke was 1.8% in patients without atherosclerotic disease of the ascending aorta, and 8.7% in patients with the disease (p < 0.0001). Diabetes mellitus was also a predictive factor (p = 0.04). A new and unique finding of this study was that the middle-lateral segment is an independent predictive factor for postoperative stroke, with a relative risk of 26% (p = 0.04).CONCLUSIONSPatients with atheromatosis in the ascending aorta had an 8.7% incidence of postoperative stroke, in spite of minor surgical modifications. The risk depended on the presence, location and extent of the disease. Randomized trials evaluating alternative surgical strategies in coronary surgery are urgently needed in high risk patients.

Summary Microprojectile bombardment of Brassica napus microspores was investigated based on transient gene expression of the marker genes β-glucuronidase (gus) and green fluorescent protein (gfp). Biological and physical bombardment parameters to maximize gene delivery were established. Microspore viability after bombardment was also determined. Evaluation of 4 different promoters, sunflower polyubiquitin, napin A, 1' and CaMV-35S led to the use of the polyubiquitin promoter for standardization of bombardment parameters. This promoter showed the highest transient gene expression in early microspore stages. Developmental stage and quality ofthe microspores were the main factors found to influence transgene expression. The highest transient expression and viability frequencies were observed after bombardment of microspores incubated for 48 h or 72 h at 32ΰC after isolation. Filtration of the 72 h-incubated microspores through cell strainers (sieves) retained only expanded and embryogenic microspores for bombardment. As a result of the enrichment of viable microspores, transient gene expression frequencies were considerably improved compared to microspores bombarded over filter papers or agarose plates. Under the best experimental conditions, transient GUS expression was observed in 0.17% of the total bombarded microspores. However, the drastic drop in microspore viability observed after bombardment was the largest drawback, resulting in a low regeneration rate of 0.06% microspore-derived embryos. The use of the marker gene, S65T-gfp, for microspore transformation is promising, since it allows detection and potential selection of viable transgenic microspores and their regeneration into embryos.

Red cell deformability is an important determinant of peripheral blood flow. In this study the red cell filtration rate (RFR) was measured in 17 patients admitted for acute embolic lower limb ischaemia. The severity of the limb ischaemia on admission was scored from 0 to 6 depending on the degree of pain, and loss of motor and sensibility functions in the ischaemic extremity. Patients who presented with very severe ischaemia on admission underwent early embolectomy whereas those with less severe ischaemia (range from 0 to approximately 3 in ischaemic score) initially were treated conservatively with heparin. Red cell filtration rate was assessed with a standard microfiltration method and expressed in microliters/sec. On admission the RFR averages 30 +/- 5 microliters/sec (mean +/- S.D.), significantly lower than the corresponding value in a normal population of similar age (50 +/- 5 microliters/sec). There was an inverse relationship (r = 0.74, P less than 0.001 by use of linear regression analysis) between the ischaemic score and the RFR on admission, with low RFR values being related to high ischaemic scores. Forty-eight hours after revascularisation, the RFR had improved significantly in the patients having undergone successful revascularisation, whereas it was unchanged in conservatively treated patients. It is concluded that patients with acute embolic lower limb ischaemia have an impaired red cell filtration rate suggesting decreased red cell deformability, the impairment being related to the severity of the ischaemia. A poor red cell filtration rate on admission is related to increased risk of postoperative gangrene and/or cardiac death. Successful revascularisation, but not conservative heparin treatment is associated with increasing deformability.

The nucleotide sequence of the early (transforming) region from the European elk papillomavirus (EEPV) double-stranded DNA has been determined together with flanking regions. The established sequence, which is 5732 bp long, shows that the genome of EEPV is closely related to the previously sequenced bovine papillomavirus type 1 (BPV-1) and deer papillomavirus (DPV) genomes. Seven open reading frames (ORFs), designated E1–E7, were identified in similar positions as in the BPV-1 genome. The E1 and E5 regions were best conserved. The strong homology between the E5 ORFs of EEPV, BPV-1 and DPV is interesting in the light of the recent proposal that these ORFs encode a major transforming function (Schiller et al., 1986; DiMaio et al., 1986). A set of mRNAs, comprising six size classes, were identified in EEPV-transformed cells. At least two different promoters appear to control EEPV transcription in transformed cells.

With freshly ejaculated semen from 15 donors, 1017 inseminations were performed in 247 women. One hundred conceptions were achieved. The degree of crystallization of the cervical mucus and the characteristics of the donor semen were examined at each insemination. One insemination only from one single ejaculate was performed in each cycle. This procedure made it possible to describe the quality of the cervical mucus and the sperm sample characteristics on each occasion of conception. The conception rate was significantly reduced (P less than 0.001) when the degree of crystallization of the cervical mucus was impaired. The conception rate was low when the sperm density was less than 20 million/ml, the percentage of abnormal sperm was above 50, and the degree of motility was graded 2 or less. The in vitro capillary sperm penetration test gave reliable information on the progressive motility of the sperm, showing a good correlation between penetration ability and conception rate.