压电基因传感器阵列的初步构建

目的　初步构建适于临床基因检测的新方法即压电基因传感器阵列。方法　制作 2× 5型基因传感器阵列 ,在阵列各检测池的银膜上通过巯基修饰法固定合成大肠杆菌肠毒素基因探针 ,而后与各靶序列进行杂交 ,以自制集成电路及电脑软件对频率数据进行采集与分析。结果　 12h内 ,气相对照检测池频率变化± 2Hz ,液相对照检测池频率变化± 4Hz ;基因探针固定后频率下降 ( 4 8± 5 )Hz ;与完全配对的靶序列杂交 ,0 .5h达到稳定状态 ,并使频率下降 ( 4 3± 5 )Hz ;与有 1～ 2个碱基错配的序列杂交 ,频率下降仅分别为 ( 2 1± 5 )、( 7± 5 )Hz ,与 3个碱基错配的序列及不相关序列杂交 ,频率无明显变化。结论　初步构建的基因传感器阵列有很好的稳定性和特异性 ,敏感性尚待进行一步提高。     

非甲至非戊型肝炎患者TTV检测和部分基因序列分析

目的：研究新近报道的一种与输血后肝炎有关的病毒在中国郑州地区正常人群、HBsAg阳性者、非甲至非戊型肝炎患者中的感染和基因序列变异情况。方法：采用TTV基因组ORF1区的套式聚合酶链反应(nested-PCR)方法对血清标本进行检测,并对非甲至非戊型肝炎患者中的TTV分离株进行序列测定。结果：TTV DNA在非甲至非戊型肝炎患者中检出率为45％(18／40);在正常体检者中检出率为17．7％(17／96);在HBsAg阳性者(ALT≤34 IU／L)中检出率22．7％(10／44)。TTV分离株序列与日本株(CLON22)相对应位置的核苷酸同源性为．1％。结论：TTV在非甲至非戊型肝炎患者TTV的感染率明显高于其他人群,可能是非甲至非戊型肝炎的主要致病因子。     

单链构象多态性分析技术检测结核杆菌利福平耐药基因

目的：研究本地区临床分离结核杆菌ｒｐｏＢ基因突变与利福平（ＲＦＰ）耐药性的关系。方法：对３６株ＲＦＰ敏感株和４４株ＲＦＰ耐药株ｒｐｏＢ基因的３２８ｂｐ的ＰＣＲ扩增产物进行单链构象多态性（Ｓｉｎｇｌｅ－ｓｔｒａｎｄ ｃｏｍｆｏｒｍａｔｉｏｎｐｏｌｙｍｏｒｐｈｉｓｍ,ＳＳＣＰ）分析。结果：３６株ＲＦＰ敏感株的ＳＳＣＰ带谱均与参考株Ｈ３７Ｒｖ相同,４４株ＲＦＰ耐药株中,２５株高度ＲＦＰ耐药株和１１株低度ＲＦＰ耐药株的ＳＳＣＰ带谱与Ｈ３７Ｒｖ带谱有差异;另８株低度ＲＦＰ耐药株的ＳＳＣＰ带谱与Ｈ３７Ｒｖ带谱相同。结论：ＳＳＣＰ分析可检测出ｒｐｏＢ基因突变,该基因突变与本地区临床分离结核杆菌对ＲＦＰ耐药有关。     

In vivo labeling of sigma receptors in mouse brain with [3H]4-phenyl-1-(4-phenylbutyl)piperidine

4-Phenyl-1-(4-phenylbutyl)piperidine(4-PPBP) is a very potent ligand for σ (Sigma) receptors. The present study was undertaken to evaluate [³H]4-PPBPas a radioligand for in vivo labeling of cerebral σ receptors. After intravenous administration of [³H]4-PPBP to mice, there is high uptake of radioactivity in the brain. The regional distribution of radioactivity in the brain 2 h after intravenous injection of [³H]4-PPBP parallels the in vitro binding of the radioligand in rat brain (pons/medulla > cerebellum ≥ prefrontal cortex ≥ parietal cortex > hypothalamus > olfactory tubercle ≥ thalamus > hippocampus > striatum). Pretreatment with haloperidol (2 mg/kg) significantly decreases the radioactivity measured in the brain 30–120 min after injection of [³H]4-PPBP. Pretreatment with unlabeled 4-PPBP or ifenprodil also significantly decreases radioactivity in the brain 2 h after injection of [³H]4-PPBP, in a dosedependent manner. The in vivo binding of [³H]4-PPBP in the brain also is significantly inhibited by SL 82.0715, BMY 14802, 1,3-di-o-tolylguanidine (DTG), and (+)-enantiomers of pentazocine, SKF 10,047, and 3-PPP, but not by the corresponding (?)-enantiomers, consistent with stereoselectivity of inhibition obtained in in vitro binding studies. In contrast, pretreatment with dizocilpine and spiperone does not inhibit in vivo binding of [³H]4-PPBP. The results indicate that [³H]4-PPBP would be a suitable radioligand for in vivo labeling of σ receptors in brain. © 1995 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America

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Homosexuality,type 1: An Xq28 phenomenon

Despite the absence of phenotypic manifestations in alternating generations characteristic of X-linked disorders, a thesis is presented that a major type of Kinsey grades 5 and 6 male homosexuality is determined by a gene in the Xq28 region. A total of 133 families in 78 kinships of male and female homosexual probands, in addition to 116 families (including those of 40 famous homosexuals) from the literature, revealed an unbalanced secondary sex ratio in the maternal generation of male, but not of female, homosexuals. On the maternal side, in this study, the ratio of all uncles to all aunts of 90 males homosexuals was 132/209, ² = 8.52, p = 0.004. On the maternal side for the total of all sources, the ratio of uncles to aunts of male homosexuals was 241/367, ² = 13.20; p < 0.0001. The male/female ratio of the total number of maternal sibships bearing homosexuals (310/628: 0.491) was a measure of fetal wastage of the mothers' male sibs: 49%. This ratio was very close to that of the total number of children born to fathers affected with any one of nine Xq28-linked male semilethal conditions (255/508: ratio 0.556); for the difference between the two populations ² = 0.859, p = 0.354. The male/female ratio of the total number of children born to female carriers of any one of these same conditions (1,232/1,062: ratio 1.16), ² = 13.8 p 0.0001, is close to that of the total number of children in homosexual sibships: 511/413, ² = 10.4, p = 0.005. Between the number of children born to Xq28 mothers and to those born of mothers of homosexuals ² = 0.581, p = 0.446. One may readily surmise that the maternal influence so often related to homosexuality may lie in the mother being a genetic carrier, with traits thereto associated. In this study, 65% of the mothers of homosexuals had no or only one live-born brother. Additional support for a genetic hypothesis is found in the occurrence of multiple instances—almost exclusively among maternal relatives—of infertility, spontaneous abortions, miscarriages, stillbirths, remaining single past age 30, and suicide. Of 109 male and 43 female homosexual index cases in the present series there were 6 instances of brother/sister homosexual sibships. Instances of homosexual parent-to-homosexual child transmission occurred as follows: one father-to-son; one father-to-daughter; one bisexual father-son; one father/mother-to-2 sons; one of mother-to-son, and one of father-to-son and father-to-bisexual daughter. There were 16 instances of presumptive transmissions from heterosexual father-to-homosexual son and 5 of heterosexual father-to-homosexual daughter. A hypothesis is proposed: Homosexuality is due to a gene at Xq28 characterized by (i) elongated cytosine-containing trinucleotide repeats upstream to translation of a gene, (ii) elongated CpG islands upstream of the trinucleotides, and (iii) cytosine methylation of CpG islands and of the cytosine-containing trinucleotides.A limited number of long tables of data, and pedigree charts, which provide the details from which this paper was developed, may be obtained on paper or on disc from the author or, for a modest fee, from the Librarian, Health Sciences Center, State University of New York at Stony Brook, Stony Brook, New York 11794.     

Stimulus duration modulates the interaction between opioids and nitric oxide in hypoxic pial artery dilation

Since recent studies show that pial artery dilation during a 20 or 40 min hypoxic exposure was less than that observed during a 5 or 10 min exposure, stimulus duration determines the nature of the vascular response to hypoxia. Decremented hypoxic pial dilation during longer exposure periods results, at least in part, from decreased release of methionine enkephalin (Met), an opioid known to contribute to dilation during hypoxia. Nitric oxide and cGMP contribute to both release and the vascular response to this opioid. The present study was designed to determine if the stimulus duration modulates the interaction between opioids and NO in hypoxic pial dilation using newborn pigs equipped with a closed cranial window. Elevation of CSF cGMP during hypoxia (Po2 approximately 35 mmHg) was dependent on stimulus duration (435+/-31, 934+/-46, 747+/-25, and 623+/-17 fmol/ml cGMP during normoxia and after 10, 20, and 40 min of hypoxia). Met-induced pial dilation during hypoxia was also stimulus duration dependent (7+/-1, 10+/-1, and 15+/-1, vs. 4+/-1, 6+/-1, and 8+/-2 vs. 2+/-1, 3+/-1, and 5+/-1% for 10(-10), 10(-8), 10(-6) M Met during normox and after 20, and 40 min of hypoxia). Additionally, the release of cGMP by Met during hypoxia was also stimulus duration dependent (1.8+/-0.1 vs. 1.6+/-0.1 vs. 1.3+/-0.1 fold change in CSF cGMP for 10(-8) M Met during normoxia and after 20 and 40 min of hypoxia). These data indicate that the diminished role of Met in pial dilation during longer hypoxic exposure periods results from a diminished capacity of this opioid to elicit dilation. Such impaired dilation is correlated with diminished stimulated cGMP release. These data also suggest that diminished CSF cGMP release during prolonged hypoxia contributes to decreased release of Met during longer hypoxic periods. Therefore, stimulus duration modulates the interaction between opioids and NO in hypoxic pial artery dilation.     

Effects of selective phosphodiesterase inhibitors on platelet-activating factor- and antigen-induced airway hyperreactivity, eosinophil accumulation, and microvascular leakage in guinea pigs

There is currently interest in the potential use of selective inhibitors of cyclic nucleotide phosphodiesterases (PDE) in the treatment of asthma. In this study we examined the effects of three selective PDE inhibitors, milrinone (PDE III), rolipram (PDE IV) and zaprinast (PDE V), on the broncoconstriction produced by antigen and histamine, the airway hyperreactivity and microvascular leakage after aerosol exposure to platelet-activating factor (PAF) and antigen, and the antigen-induced eosinophil infiltration in guinea-pig lung. Inhaled rolipram (0.01–10 mg ml^–1) inhibited dose dependently the bronchospasm produced by aerosol antigen (5 mg ml^–1) an anaesthetised, ventilated guinea-pigs. Rolipram (10 mg ml^–1) produced maximal inhibition of antigen-induced bronchoconstriction but only partial inhibition of the response to aerosol histamine (1 mg ml^–1). Milrinone and zaprinast (each 10 mg ml^–1) showed weak, or no, inhibitory effects against bronchoconstriction produced by aerosol antigen or histamine. Pretreatment with rolipram (10 mg kg^–1, i.p.) prevented airway hyperreactivity to histamine which develops 24 h after exposure of conscious guinea-pigs to aerosol PAF (500 g ml^–1) or antigen (5 mg ml^–1). The pulmonary eosinophil infiltration obtained with 24 h of antigen-exposure was inhibited by rolipram. In contrast, milrinone and zaprinast (each 10 mg kg^–1, i.p.) failed to reduce either the airway hyperreactivity of the eosinophil accumulation in these animals. Rolipram (1–10 mg ml^–1) reduced the extravasation of Evans blue after aerosol PAF (500 g ml^–1) at all airway levels while a lower dose (0.1 mg ml^–1) was only effective at intrapulmonary airways. Rolipram (0.01–1 mg ml^–1) markedly reduced airway extravasation produced by inhaled antigen (5 mg ml^–1). Zaprinast (1–10 mg ml^–1) was also effective against airway microvascular leakage produced by aerosol PAF or antigen while milrinone (10 mg ml^–1) had no antiexudative effect. These data support previous suggestions that pharmacological inhibition of PDE IV results in anti-spasmogenic and anti-inflammatory effects in the airways and may be useful in the treatment of asthma.     

Improvement of brain single photon emission tomography (SPET) using transmission data acquisition in a four-head SPET scanner

Attenuation coefficient maps (-maps) are a useful way to compensate for non-uniform attenuation when performing single photon emission tomography (SPET). A new method was developed to record single photon transmission data and a-map for the brain was produced using a four-head SPET scanner. Transmission data were acquired by a gamma camera opposite to a flood radioactive source attached to one of four gamma cameras in the four-head SPET scanner. Attenuation correction was performed using the iterative expectation maximization algorithm and the-map. Phantom studies demonstrated that this method could reconstruct the distribution of radioactivity more accurately than conventional methods, even for a severely non-uniform-map, and could improve the quality of SPET images. Clinical application to technetium-99m hexamethylpropylene amine oxime (HMPAO) brain SPET also demonstrated the usefulness of this method. Thus, this method appears to be promising for improvement in the image quality and quantitative accuracy of brain SPET.This work was presented in part at the World Congress on Medical Physics and Biomedical Engineering, 7–12 July 1991, Kyoto, Japan     

Effects of metoprolol on action potential and membrane currents in guinea-pig ventricular myocytes

Summary The effects of the beta-adrenoceptor antagonist metoprolol on action potentials and membrane currents were studied in single guinea-pig ventricular myocytes. The experiments were carried out using the nystatin-method of whole-cell technique. This method was used in order to prevent the run-down of the calcium current. Metoprolol at concentrations of 10–100 mol/l shortened action potential in a dose-dependent way. The drug only decreased resting membrane potential at a concentration of 100 mol/1 in two out of five cells. Under voltage-clamp conditions, metoprolol blocked the high threshold calcium current at concentrations of 30 and 100 mol/l to 82 ± 4% and 73 ± 5% from control, respectively. The drug decreased the inward rectifying potassium current in a concentration-dependent manner. This effect was evident for inward current at voltages negative to the apparent reversal potential and for outward current at voltages between –30 and –80 mV. This blocking effect on the inward rectifying potassium current can explain the effect on resting membrane potential. At voltages positive to –30 mV metoprolol increased a time-independent outward current. This metoprolol-enhanced outward current was blocked by barium and cesium. This result suggests that the metoprolol-enhanced current is carried by potassium. The current component enhanced by metoprolol was not sensitive to glibenclamide and tetraethylammonium applied externally, which suggests that the adenosine triphosphate-sensitive channel is not the target of metoprolol. The activation of this time-independent outward current by metoprolol and the blocking effects on the calcium current seem to explain the shortening in action potential induced by the drug. Send offprint requests to J. Sánchez-Chapula at the above address     

Modulation of the cutaneous responsiveness of neurones in the primary somatosensory cortex during conditioned arm movements in the monkey

Summary The present experiments were designed to investigate the neuronal mechanisms, at the level of the primary somatosensory cortex, which underlie the observation that somatosensory cortical potentials evoked by air puff stimuli directed at the forearm are decreased, in a nonspecific and widespread manner, during voluntary movements about the elbow. Unitary discharge was recorded from 131 cells receiving cutaneous input from the hairy skin of the forearm or hand (areas 3b and 1) of two monkeys trained to perform rapid movements of the contralateral arm (elbow flexion or extension). Evoked unitary responses to air puff stimuli applied to the centre of the cell's receptive field, at various delays before and after the onset of movement, were recorded. Movement produced a significant decrease in the short latency excitatory response to the air puff in 89% of the cells (117/131); the remaining 11% were not modulated by movement. This movement-related gating of cutaneous inputs occurred regardless of the response pattern of the cells to movement alone, being observed in 91% of the cells with no movement-related discharge, and 89% of those with movement-related discharge. The air puff responses of cells with inputs from the forearm and the dorsum of the hand were all similarly modulated by movement and the modulation was clearly present prior to the onset of movement (mean onset, -66 ms). Variation in the depth of modulation as a function of the direction of the movement, flexion or extension, was observed in only a very small proportion of the modulated units (16/117); most showed no relationship to direction. It is suggested that, in this experimental situation, much of the modulation appears to occur at a pre-cortical level since there was no relationship between the pattern of discharge of cells in relation to movement alone and the pattern of movement-related gating of their responses to the air puff. Effects which might be consistent with a cortical origin for the modulation were only infrequently observed. The present results are strikingly similar to those obtained using the evoked potential method, and thus support the hypothesis that, in this task of rapid elbow movements, movement modulates the transmission of cutaneous signals from the hairy skin of the distal forelimb to primary somatosensory cortex in a nonspecific and widespread fashion.