Human influenza virus hemagglutinin with high sensitivity to proteolytic activation.

To examine the prerequisites for cleavage activation of the hemagglutinin of human influenza viruses, a cDNA clone obtained from strain A/Port Chalmers/1/73 (serotype H3) was subjected to site-directed mutagenesis and expressed in CV-1 cells by using a simian virus 40 vector. The number of basic residues at the cleavage site, which consists of a single arginine with wild-type hemagglutinin, was increased by inserting two, three, or four additional arginines. Like wild-type hemagglutinin, mutants with up to three additional arginines were not cleaved in CV-1 cells, but insertion of four arginines resulted in activation. When the oligosaccharide at asparagine 22 of the HA1 subunit of the hemagglutinin was removed by site-directed mutagenesis of the respective glycosylation site, only three inserted arginines were required to obtain cleavage. Mutants containing a series of four basic residues were also generated by substituting arginine for uncharged amino acids immediately preceding the cleavage site. The observation that these mutants were not cleaved, even when the carbohydrate at asparagine 22 of HA1 was absent, underscores the fact that the basic peptide had to be generated by insertion to obtain cleavage. The data show that the hemagglutinin of a human influenza virus can acquire high cleavability, a property known to be an important determinant for the pathogenicity of avian influenza viruses. Factors important for cleavability are the number of basic residues at the cleavage site, the oligosaccharide at asparagine 22, and the length of the carboxy terminus of HA1.     

Effect of high altitude on sensitivity to the taste of phenylthiocarbamide

Sensitivity to the taste of phenylthiocarbamide (PTC) was studied using the Harris-Kalmus method in healthy human volunteers at sea level and then subsequently at an altitude of 3500 m over a period of 3 weeks, after which they were brought back to sea level. Blood sugar, insulin and blood cortisol levels were estimated weekly. The results indicated that, out of 51 subjects studied, 26 (55%) were PTC tasters at sea level. Eight of those unable to taste PTC at sea level tested as tasters at high altitude, and 2 of them reverted to being non-tasters on return to sea level. In the blood, an increase in cortisol and blood insulin levels was seen without any significant change in sugar levels. All the changes recorded at high altitude tended to return to basal values after re-induction to sea level. The study suggests that high-altitude hypoxia in some way, possibly involving changes in hormonal profile among other factors, causes an alteration in sensitivity to the taste of PTC, resulting in some of the individuals shifting to lower PTC sensitivity. Received: 24 November 1999 / Accepted: 25 January 2000     

一种基于数字免疫原理的伤寒沙门菌高灵敏度检测方法

目的 设计一种基于数字化免疫显色原理的检测方法,实现伤寒沙门菌的高灵敏度快速检测.方法 以伤寒沙门菌为检测对象,选取氨基磁珠对目标细菌进行富集,以偶联特异性抗体的量子点对目标细菌进行特异性荧光标记;设计制作反应腔室,在液相条件下完成对目标细菌的富集和特异性荧光标记;通过荧光显微成像和荧光特征自动提取,实现对目标细菌的"...     

A new approach to high sensitivity differential hybridization

We describe a new approach to differential hybridization, designed to identify cDNA clones representing rare mRNA species. Duplicate filters carrying a library of cDNA from phorbolmyristate acetate (PMA)-induced EL-4 cells in λgt11 were hybridized with high concentrations of unlabeled, cloned, single-stranded cDNA from induced and control EL-4 cells, respectively. Plaques binding single-stranded cDNA were revealed by a second round of hybridization with ³⁵S-labeled DNA complementary to the vector moiety of the single-stranded cDNA. Plaques corresponding to PMA-induced mRNAs occurring at a level of about 1 part in 15000 were isolated. We believe the method is at least ten times more sensitive than conventional differential hybridization.     

Enhanced mitochondrial sensitivity to creatine in rats bred for high aerobic capacity.

Qualitative and quantitative measures of mitochondrial function were performed in rats selectively bred 15 generations for intrinsic aerobic high running capacity (HCR; n = 8) or low running capacity (LCR; n=8). As estimated from a speed-ramped treadmill exercise test to exhaustion (15 degrees slope; initial velocity of 10 m/min, increased 1 m/min every 2 min), HCR rats ran 10 times further (2,375+/-80 m) compared with LCR rats (238+/-12 m). Fiber bundles were obtained from the soleus and chemically permeabilized. Respiration was measured 1) in the absence of ADP, 2) in the presence of a submaximally stimulating concentration of ADP (0.1 mM ADP, with and without 20 mM creatine), and 3) in the presence of a maximally stimulating concentration of ADP (2 mM). Although non-ADP-stimulated and maximally ADP-stimulated rates of respiration were 13% higher in HCR compared with LCR, the difference was not statistically significant (P>0.05). Despite a similar rate of respiration in the presence of 0.1 mM ADP, HCR rats demonstrated a higher rate of respiration in the presence of 0.1 mM ADP+20 mM creatine (HCR 33% higher vs. LCR, P<0.05). Thus mitochondria from HCR rats exhibit enhanced mitochondrial sensitivity to creatine (i.e., the ability of creatine to decrease the Km for ADP). We propose that increased respiratory sensitivity to ADP in the presence of creatine can effectively increase muscle sensitivity to ADP during exercise (when creatine is increased) and may be, in part, a contributing factor for the increased running capacity in HCR rats.     

Observational evidence of the sensitivity of some fish species to environmental stressors in Mediterranean rivers

We describe a tolerance classification based on the physicochemical parameters of water and habitat quality indices for native and introduced fish species in northeastern Spain. Although fish are widely used as biological indicators, the tolerance categories currently available for Spanish Mediterranean fish species are uncertain or subjective. We sampled 430 sites covering a wide range of river conditions, allowing us to develop tolerance scores based on a quantitative assessment. We used indirect ordination methods to establish stress gradients between the environmental variables compiled during the monitoring surveys carried out from 2002 to 2008. We then drew up a tolerance classification for the most common species in our dataset using weighted average inference models based on the following gradients: water quality and the state of physical habitat. The results were corroborated by multivariate gradient analysis. The fish scores obtained were applied to an independent dataset and a positive correlation was found between assessment protocols. Our results corroborate the use of fish, including exotic fish species, as biological indicators. Our findings may be used in a regional Index of Biotic Integrity and for the use of fish in bioassessment studies based on scores.     

Spectral sensitivity of the African cichlid fish,Haplochromis burtoni

Summary Spectral sensitivity of the cichlid fishHaplochromis burtoni was measured under both scotopic and photopic conditions using a two-choice, food reward, operant conditioning paradigm. The highest absolute sensitivity (scotopic) is one quantum for every 5 to 50 rods measured at 475 nm (equivalent to a corneal irradiance of 3.8×10⁶ Q s^–1 cm^–2). A P500₁ photopigment apparently mediates spectral sensitivity over most of the visible spectrum; microspectrophotometric studies of rods had previously shown them to contain this photopigment. However, the scotopic behavioral action spectrum shows a sensitivity to short wavelength light higher than is consistent with a P500₁ photopigment alone mediating the scotopic visual process. Determinations made under photopic conditions reveal a behavioral action spectrum broader than that found under scotopic conditions and consistent with mediation by interaction of the three known cone types in an opponent processing manner. The calculated photopic threshold value of approximately 10⁴ Q s^–1 (receptor)^–1 is in agreement with results from other species and corresponds to a corneal irradiance of about 7×10¹⁰Q s^–1cm^–2.     

Photopic spectral sensitivity of a teleost fish,the roach (Rutilus rutilus), with special reference to its ultraviolet sensitivity

Summary This study reports photopic spectral sensitivity curves (351–709 nm) for four individual roach,Rutilus rutilus, determined by two choice appetitive training. All four curves show four sensitivity maxima at 361–398 nm, 421–448 nm, 501–544 nm and 634–666 nm which are related to the four known roach photopic visual pigments (Avery et al. 1982). The overall shape of the curves at long wavelengths indicates inhibitory interactions between the red and green cone mechanisms. That the high behavioural sensitivity in the UV is caused by a specific ultraviolet visual pigment and is not due to aberrant stimulation of the other cone types is shown by the redetermination of spectral sensitivity at short wavelengths (351–501 nm) following the selective bleaching of the three longer wavelength visual pigments. This depresses the blue sensitivity to a greater degree than the relatively unaffected UV sensitivity maximum. Spectral transmission data from two corneas and four lenses show that they transmit considerable amounts of light in the near UV.     

Trait structure and redundancy determine sensitivity to disturbance in marine fish communities

Trait diversity is believed to influence ecosystem dynamics through links between organismal traits and ecosystem processes. Theory predicts that key traits and high trait redundancy—large species richness and abundance supporting the same traits—can buffer communities against environmental disturbances. While experiments and data from simple ecological systems lend support, large‐scale evidence from diverse, natural systems under major disturbance is lacking. Here, using long‐term data from both temperate (English Channel) and tropical (Seychelles Islands) fishes, we show that sensitivity to disturbance depends on communities’ initial trait structure and initial trait redundancy. In both ecosystems, we found that increasing dominance by climatically vulnerable traits (e.g., small, fast‐growing pelagics/corallivores) rendered fish communities more sensitive to environmental change, while communities with higher trait redundancy were more resistant. To our knowledge, this is the first study demonstrating the influence of trait structure and redundancy on community sensitivity over large temporal and spatial scales in natural systems. Our results exemplify a consistent link between biological structure and community sensitivity that may be transferable across ecosystems and taxa and could help anticipate future disturbance impacts on biodiversity and ecosystem functioning.     

Mechanisms of high sensitivity of Mongolian pika spp. (Pallas) to Yersinia pestis

The regularities of the infectious process of plague in a sensible Pallas' pika species, are presented. The specific infectious process was shown to pass a number of phases. These phases are characterized by certain changes in the composition of the leukocytic pool of the peripheral blood, the immune system and the activity of the bactericidal systems of professional phagocytes. The infectious process in sensitive animals is accompanied by the formation of secondary relative immunodeficiency with a pronounced hypersuppressive component, which leads to disturbances in the cooperative interaction of immunocompetent cells; in its turn, it results in dysbalance in the cytokine system and, as a consequence, to irreversible disturbances in homeostasis, leading to the death of the infected animal.     

Selective determination of fish aspartate aminotransferase isoenzymes by their differential sensitivity to proteases

Various proteases (proteinase K, subtilisin, trypsin and chymotrypsin) were used to study the selective inactivation of the aspartate aminotransferase (EC 2.6.1.1) isoenzymes of grey mullet (Mugil auratus Risso; Osteichthyes). The cytosolic isoenzyme was significantly inactivated by proteinase K, subtilisin and chymotrypsin, while the mitochondrial isoenzyme was sensitive only to proteinase K and to high doses of trypsin. Further identification of the aspartate aminotransferase isoenzymes was based on their discrete sensitivity toward chymotrypsin. Chymotrypsin (1 mg/ml) successfully inhibited purified cytosolic aspartate aminotransferase as well as cytosolic isoenzyme from plasma, whereas the mitochondrial form persisted unaffected. Similar results were obtained when examining liver and red muscle homogenates. This method revealed that the increased total activity of aspartate aminotransferase in fish plasma with induced acute liver injury, was partially a result of the mitochondrial isoenzyme leakage from damaged tissue.     

Chemoreceptor sensitivity and maladaptation to high altitude in man

Studies were carried out to find out the role of chemoreceptor sensitivity in the causation of maladaptation syndromes on acute exposure to altitude. The experiments were done in two phases. In phase I, the responses in chemoreceptor sensitivity were studied in altitude acclimatized subjects and compared with those who suffered from either High Altitude Pulmonary Oedema (HAPO) or Acute Mountain Sickness (AMS). In Phase II, a similar comparison was done in two groups of subjects, one representing normal sojourners at 3,500 m and the other being subjects who had just recovered from HAPO. The first phase was done at Delhi; and the second at an altitude of 3,500 m. Parameters of assessment were hypoxic sensitivity, carbon dioxide sensitivity, ventilation (VE), respiratory frequency (Rf), forced vital capacity (FVC), forced expiratory volume at the first second (FEV1), heart rate (HR), blood pressure (BP), and oral temperature (Tor). The results showed significantly lower sensitivity to both hypoxia and carbon dioxide in maladapted subjects, as compared to those who were acclimatized in both the categories suggesting thereby that reduced chemoreceptor sensitivity might be an initiating factor in the causation of maladaptation syndromes at altitude.