Hippocampal noradrenergic responses to CRF injected into the locus coeruleus of unanesthetized rats

Intracerebral administration of corticotropin-releasing factor (CRF) activates cerebral noradrenergic neurons. Direct infusion of CRF into the locus coeruleus (LC) increases norepinephrine (NE) release in the cortex and hippocampus as assessed by in vivo microdialysis. In a recent study using in vivo chronoamperometry in anesthetized rats, CRF injected into the LC increased apparent NE release in the hippocampus, but did so after a significant delay, much longer than observed following infusion of glutamate into the same site. Because this delay may have been an artifact of the urethane anesthesia, we developed a method for chronoamperometric recording from the hippocampus of unanesthetized rats. CRF infusion into the LC of such animals induced an increase in the apparent release of hippocampal NE after a mean delay of about 7 min, reached a peak around 16 min after CRF, and dissipated within 30 min. Thus the response closely resembled that previously reported in urethane-anesthetized rats. As in anesthetized rats, glutamate infused into the same site resulted in a much more rapid response (starting within 1 min and with a peak around 7 min). These results suggest that the urethane anesthesia does not substantially alter hippocampal NE release following infusion of CRF into the LC, and that the relatively long delay in the response is not an artifact of the anesthesia. The large differences in the responses to glutamate and CRF suggest that the effects of CRF are not exerted directly on receptors on LC neurons, and more likely reflect indirect actions on other cells in this region.     

The supraclavicular block with a nerve stimulator: to decrease or not to decrease, that is the question

Portable nerve stimulators for nerve blocks have been available for more than 40 yr. It is generally accepted that seeking a motor response at low outputs increases the chances of success. It is customary to start the procedure at a higher current with the goal of finding the nerve. After an adequate response is elicited, the current is decreased before the local anesthetic is injected. However, how low is low enough and, for that matter, how high is too high have not been adequately determined. Our experience seems to indicate that, in the supraclavicular block, the type of response obtained is as important as the output at which it is elicited, provided that this current is not higher than 1 mA. In this context, it is theoretically possible that our initial seeking current of 0.9 mA could be an adequate injection current if it is combined with an appropriate response. We designed this study to test the hypothesis that a response of the fingers in flexion or extension, elicited at 0.9 mA, could be followed immediately by the local anesthetic injection. We did not intend to compare 0.5 and 0.9 mA as minimum stimulating currents but rather as currents able to elicit an unmistakable motor twitch. Sixty patients were randomly assigned to one of two groups. Group 1 (n = 30) was injected with a motor twitch in the fingers that was still visible at 0.5 mA. Group 2 (n = 30) was injected after a similar response to that in Group 1 was elicited, but at the initial output of 0.9 mA, without any further decrease. The blocks were injected with 40 mL of local anesthetic solution. One patient was excluded from the study for failing to meet protocol criteria. The success rate in the remaining 59 patients was 100%; success was defined as complete sensory blockade at the median, ulnar, and radial nerve territories of the hand that was accomplished in 相似文献   

Engineering a reduced Escherichia coli genome

Our goal is to construct an improved Escherichia coli to serve both as a better model organism and as a more useful technological tool for genome science. We developed techniques for precise genomic surgery and applied them to deleting the largest K-islands of E. coli, identified by comparative genomics as recent horizontal acquisitions to the genome. They are loaded with cryptic prophages, transposons, damaged genes, and genes of unknown function. Our method leaves no scars or markers behind and can be applied sequentially. Twelve K-islands were successfully deleted, resulting in an 8.1% reduced genome size, a 9.3% reduction of gene count, and elimination of 24 of the 44 transposable elements of E. coli. These are particularly detrimental because they can mutagenize the genome or transpose into clones being propagated for sequencing, as happened in 18 places of the draft human genome sequence. We found no change in the growth rate on minimal medium, confirming the nonessential nature of these islands. This demonstration of feasibility opens the way for constructing a maximally reduced strain, which will provide a clean background for functional genomics studies, a more efficient background for use in biotechnology applications, and a unique tool for studies of genome stability and evolution.     

一种新的免疫抑制剂FTY720对小鼠心脏移植的救治作用(英文)

AIM: FTY720 is a new synthetic immunosuppressive agent which has a unique mechanism of action and induces long-term graft acceptance in rat and dog allotransplantation as prophylactic administration. The present study investigated whether FTY720 was able to rescue ongoing acute rejection of solid organ transplants in a mouse heterotopic cardiac transplantation model. METHODS: BALB/c hearts were heterotopically grafted in C57BL/6 mice. FTY720, at the doses of 0.5, 1, and 5 mg·kg~(-1)·d~(-1) or vehicle was administered to recipients once daily by oral gavage from d 3 to d 7 after transplantation. Histological changes of grafts, and the lymphocyte number in the peripheral blood and the peripheral lymph nodes were determined on d 5 after transplantation. RESULTS: FRY720 prolonged the median graft survival time dose-dependently and significantly. Histological evaluation revealed less lymphocytic infiltration in cardiac allografts treated with FTY720. Moreover, FTY720 remarkably lowered the number of periph     

Effect of UVA light on the activity of several aged human lens enzymes

PURPOSE: To determine the effect of UVA irradiation on the specific activities of several protective and metabolic enzymes in aged human lenses. METHODS: Intact human lenses (ages 55-75) in artificial aqueous humor were irradiated in a quartz cuvette with UVA light (925 J/cm(2) per hour) at +20 degrees C. The lenses were homogenized and the activities of enzymes in the water-soluble (WS) fraction were measured in irradiated and nonirradiated lenses. RESULTS: One hour of UVA photolysis of human lens resulted in a 70% loss in glutathione reductase (GR)-specific activity and a 24% loss in glyceraldehyde-3-phosphate dehydrogenase (G3PD)-specific activity. At the same time, glutathione peroxidase (GPx) and superoxide dismutase (SOD) showed little or no loss in specific activity. GR and G3PD showed similar losses when human lenses were photolyzed with the same dose of UVA light delivered to the lens over 8 hours, using a 12.5% neutral-density filter (ndf), or over 24 hours with a 4.25% ndf. One hour photolysis of the human lens WS fraction under anaerobic conditions yielded an almost complete inactivation of GR, but only an 18% loss of G3PD activity. Under aerobic conditions, however, both enzyme activities were almost completely lost. Clear fetal bovine lenses, photolyzed under the identical conditions, displayed essentially the same loss of GR activity. CONCLUSIONS: UVA light causes inactivation of GR in human and fetal calf lenses under both anaerobic and aerobic conditions. This suggests that flavine adenine dinucleotide (FAD), the prosthetic group of GR, may be responsible for the enzyme's self-sensitizing properties. WS proteins from aged human lens generate reactive oxygen species (ROS) during UVA irradiation, which may be responsible for the inactivation of G3PD.     

The effect of UVA light on the anaerobic oxidation of ascorbic acid and the glycation of lens proteins

PURPOSE: To determine whether UVA-excited human lens chromophores can cause the oxidation of ascorbic acid in the absence of oxygen, and whether these oxidation products are capable of glycating lens proteins. METHODS: The oxidation of ascorbic acid, mediated by UVA irradiation in the presence of aged human lens proteins, was measured in the absence of oxygen by the decrease in absorbance at 265 nm in vitro. An action spectrum from 320 to 400 nm was determined for both ascorbate oxidation and the photobleaching of the lens yellow pigments at lambda = 350 nm. The UVA-mediated oxidation products of [U-(14)C]ascorbate were quantified by HPLC. Glycation was assayed by the UVA-dependent incorporation of [U-(14)C]ascorbate into lens proteins with a water-insoluble (WI) fraction in vitro, with incubated whole human lenses, and with a WI fraction after a 5- to 7-day exposure to ambient sunlight. An enzymatic digest of [U-(14)C]ascorbate-labeled proteins was fractionated over HPLC columns and compared with the 330-nm absorbance profile of a proteolytic digest of aged human lens proteins. RESULTS: Aged human lens WI proteins absorbed UVA light (86 J/h per square centimeter) and oxidized 33 to 45 nanomoles of ascorbate over 1 hour in the absence of oxygen. No ascorbate oxidation was detected, however, in the dark control. An action spectrum showed that ascorbate oxidation occurred throughout the UVA region, with lambda(max) at 350 nm, which was similar to the action spectrum obtained for the photobleaching of the lens chromophores. Anaerobic UVA irradiation of aged human lens proteins for 2 hours with [U-(14)C]ascorbate resulted in a 40% loss of ascorbate with the accumulation of dehydroascorbic acid, diketogulonic acid, and oxalate. After subsequent incubation for 24 hours, the ascorbate oxidation products disappeared, with a corresponding incorporation of radioactivity into lens proteins. Chromatography of enzymatic digests of the labeled proteins produced peaks that coeluted with several of the 330-nm absorbing peaks in an aged human lens protein digest. Irradiation of whole human lenses for 2 hours caused a 33% loss of total lens ascorbate. UVA irradiation of aged human lenses for 2 hours resulted in the incorporation of ascorbate into lens proteins during the ensuing 24 hours in the dark. Exposure of aged human lens WI proteins to reflected ambient sunlight (1.1 J/h per square centimeter) for 5 to 7 days in the absence of oxygen also produced an increased incorporation of [(14)C]ascorbate into protein when compared with dark control samples. CONCLUSIONS: These data argue that UVA light can cause an oxidation of ascorbic acid in the absence of oxygen, due to the activation of the sensitizers present in aged human lens WI proteins. The oxidation products formed were the same as those seen in the presence of oxygen, and were rapidly incorporated into protein, apparently by Maillard-type chemistry. These data argue that ascorbate glycation can occur under the low oxygen levels thought to exist in the human lens nucleus in vivo.     

Polycomplexes of poly(acrylic acid) with streptomycin sulfate and their antibacterial activity.

Complex formation between streptomycin sulfate and poly(acrylic acid) has been studied in aqueous solutions by turbidimetric, potentiometric and viscometric methods as well as by FTIR spectroscopy. It was shown that these polycomplexes are stabilized by electrostatic interactions. The solubility of polycomplexes was examined as a function of pH and it was found that at pH values below 3.1 the polycomplexes undergo complete dissociation or dissolution. The antimicrobial activity of the drug and its polycomplex was evaluated using Sarcina sp. as a model organism. It was demonstrated that the polycomplexes have an antimicrobial activity on the same level as the free drug.     

Ambulatory blood pressure and arterial stiffness web‐based telemonitoring in patients at cardiovascular risk. First results of the VASOTENS (Vascular health ASsessment Of The hypertENSive patients) Registry

The VASOTENS Registry is an international telehealth‐based repository of 24‐hour ambulatory blood pressure monitorings (ABPM) obtained through an oscillometric upper‐arm BP monitor allowing combined estimation of some vascular biomarkers. The present paper reports the results obtained in 1200 participants according to different categories of CV risk. Individual readings were averaged for each recording and 24‐hour mean of brachial and aortic systolic (SBP) and diastolic blood pressure (DBP), pulse wave velocity (PWV), and augmentation index (AIx) obtained. Peripheral and central BP, PWV and AIx values were increased in older participants (SBP only) and in case of hypertension (SBP and DBP). BP was lower and PWV and AIx higher in females. PWV was increased and BP unchanged in case of metabolic syndrome. Our results suggest that ambulatory pulse wave analysis in a daily life setting may help evaluate vascular health of individuals at risk for CV disease.     

Hydration of Cement in the Presence of Biocidal Modifiers Based on Metal Hydrosilicates

This article presents the results of a study of the characteristics of hydration and properties of a composite biocidal cement binder containing hydrosilicates of barium, copper or zinc. It was found that copper hydrosilicates block hydration processes, and when zinc hydrosilicates are used, the rate of hydration is determined by the content of silicic acid. The limiting concentrations of biocidal modifiers have been established: zinc hydrosilicates—no more than 4% and copper hydrosilicates—no more than 0.5%, which are advisable to use for the manufacture of a biocidal composite binder. It is shown that modifying additives slow down the setting time, the amount of tricalcium silicate in cement stones increases, and their strength for some compositions decreases. Active binding of portlandite with the formation of calcium hydrosilicates occurs when the content of zinc hydrosilicates is 2%, which leads to an increase in the strength of the materials.     

Genetic Diversity of Porcine Circovirus 2 in Wild Boar and Domestic Pigs in Ukraine

Porcine circovirus type 2 (PCV2) is responsible for a number of porcine circovirus-associated diseases (PCVAD) that can severely impact domestic pig herds. For a non-enveloped virus with a small genome (1.7 kb ssDNA), PCV2 is remarkably diverse, with eight genotypes (a–h). New genotypes of PCV2 can spread through the migration of wild boar, which are thought to infect domestic pigs and spread further through the domestic pig trade. Despite a large swine population, the diversity of PCV2 genotypes in Ukraine has been under-sampled, with few PCV2 genome sequences reported in the past decade. To gain a deeper understanding of PCV2 genotype diversity in Ukraine, samples of blood serum were collected from wild boars (n = 107) that were hunted in Ukraine during the November–December 2012 hunting season. We found 34/107 (31.8%) prevalence of PCV2 by diagnostic PCR. For domestic pigs, liver samples (n = 16) were collected from a commercial market near Kharkiv in 2019, of which 6 out of 16 (37%) samples were positive for PCV2. We sequenced the genotyping locus ORF2, a gene encoding the PCV2 viral capsid (Cap), for 11 wild boar and six domestic pig samples in Ukraine using an Oxford Nanopore MinION device. Of 17 samples with resolved genotypes, the PCV2 genotype b was the most common in wild boar samples (10 out of 11, 91%), while the domestic pigs were infected with genotypes b and d. We also detected genotype b/d and b/a co-infections in wild boars and domestic pigs, respectively, and for the first time in Ukraine we detected genotype f in a wild boar from Poltava. Building a maximum-likelihood phylogeny, we identified a sublineage of PCV2 genotype b infections in both wild and domestic swine, suggesting a possible epizootic cluster and an ecological interaction between wild boar and domestic pig populations in northeastern Ukraine.