The hemodynamic effects of AT-112, an analog of ketanserin, in portal hypertensive rats

Proteins that are actively secreted by Mycobacterium tuberculosis generate immune responses in the infected host. This has prompted the characterization of protein components of mycobacterial culture filtrates to develop subunit vaccines and immunodiagnostic reagents. Fractionation of filtrates of M. tuberculosis cultures has yielded an abundant protein called MPT63, which has an apparent molecular mass of 18 kDa. We report the molecular cloning and nucleotide sequence of the mpt63 gene, purification of recombinant MPT63 antigen from Escherichia coli cells, and serological characterization of MPT63. Nucleotide sequence analysis of mpt63 identified an open reading frame encoding a protein of 159 amino acids (aa) consisting of a 29-aa secretion signal peptide and a 130-aa mature MPT63 protein. Recombinant MPT63 protein, purified from E. coli cells, and native MPT63, purified from M. tuberculosis culture filtrates, were indistinguishable in serological assays. Thus, the recombinant protein constitutes a valuable reagent for immunological studies. MPT63 evoked humoral immune responses in guinea pigs infected with virulent M. tuberculosis by the aerosol route. The mpt63 gene is found only in species of the M. tuberculosis complex, as shown by DNA hybridization experiments. Moreover, polyclonal antibody against MPT63 does not cross-react with proteins of a common environmental mycobacterial species, Mycobacterium avium. The absence of cross-reactive epitopes makes MPT63 an attractive candidate as an M. tuberculosis complex-specific diagnostic reagent. In particular, evaluation of MPT63 as an M. tuberculosis complex-specific reagent for diagnostic skin testing is under way.     

微型计算机在微粒检测中的应用

微型计算机在微粒检测中的应用＊周鑫玲曲丹丹俞岳何阳（天津大学天津３０００７２）１引言注射药物普鲁卡因青霉素的颗粒大小和分布状态是衡量该药物质量的主要内容之一。采用ＰＣ微型计算机控制和库尔特计数原理设计的ＷＪ－１Ａ型智能微粒检测仪，可用于测量普鲁卡因青...     

Postoperative epidural infusion of morphine and bupivacaine is safe on surgical wards. Organisation of the treatment, effects and side-effects in 2000 consecutive patients

BACKGROUND: There was an obvious need to improve the quality and safety of our postoperative pain treatment and to introduce an improved routine service on surgical wards. METHODS: It was decided to use postoperative epidural infusion of morphine 0.04 mg/ml and bupivacaine 2.5 mg/ml (0.25%) 4-8 ml/h as pain relief after major surgery. An education programme was run emphasising the benefits, side-effects and the importance of regular monitoring of pain intensity, vital functions (respiratory rate, blood pressure, heart rate), motor function of the legs and the need for additional drugs in order to detect side-effects as well as lack of adequate analgesic effect. A detailed observation sheet was used collecting information every 2 h throughout the epidural treatment period in order to secure monitoring and adjustment of the treatment. Close contact was maintained with the wards. RESULTS: We present a detailed analysis of our first 2000 postoperative patients, mainly after orthopaedic (46.1%), gastrointestinal (32.0%), urologic (8.7%) and vascular (8.5%) surgery. Duration of the treatment was less than 24 h in 41.4% and more than 48 h in 29.7%. Pain relief was adequate in most patients, best after vascular surgery in the lower extremities (mean VAS 0.15/10.0 (95% confidence interval 0.09-0.23)) and less after gastrointestinal (mean VAS 0.49/10 (0.43-0.54)) and thoracic surgery (mean VAS 0.59/10 (0.38-0.81)). The infusion was stopped due to respiratory depression in 3 patients (0.15%). Four (0.2%) had systolic blood pressure < 80 mmHg and had to be treated with vasopressors. A total of 56 (2.8%) patients were considered to be problem patients due to excessive sedation (0.4%), hypotension (0.7%), respiratory depression (1.6%) or lower extremity paralysis (0.05%). All patients had urinary catheter until 6 h after termination of the epidural treatment. One patient had the epidural catheter accidentally placed subarachnoidally and experienced severe respiratory depression. No permanent sequelae were recorded in the postoperative patients, but 2 traumatised patients developed epidural abscesses after 3 weeks of treatment, which resulted in lower extremity paralysis. Late response to the warning signs might have contributed to the irreversible paraplegia. CONCLUSION: Our experience with this postoperative epidural analgesia regime is favourable. It has been easy to administer and monitor. Pain relief was excellent, side-effects were few and picked up by the established routines followed by the ward staff except in the 2 trauma patients who developed epidural abscesses. The staff on the surgical wards were motivated for this kind of work. Education and strict surveillance routines are mandatory in order to secure prompt action when warning signs develops (e.g. lower limb paralysis).     

Experimental erbium: YAG laser photoablation of trabecular meshwork in rabbits: an in-vivo study

Photoablative laser trabecular surgery has been proposed as an outflow-enhancing treatment for open-angle glaucoma. The aim of the study was to investigate the time course of repair response following low-thermal Erbium: YAG laser trabecular ablation. In 20 anaesthetized rabbits gonioscopically controlled ab-interno photoablation of the ligamenta pectinata and underlying trabecular meshwork (TM) was performed with a single-pulsed (200 microseconds) Erbium: YAG (2.94 microns) laser. The right eye received 12-15 single laser pulses (2 mJ) delivered through an articulated zirconium fluoride fiberoptic and a 200 microns (core diameter) quartz fiber tip, the left unoperated eye served as control. At time intervals of 30 minutes, 2, 10, 30, and 60 days after laser treatment, eyes were processed for light- and scanning electron microscopy. The applied energy density of 6-4 J cm-2 resulted in visible dissection of the ligamenta pectinata and reproducible microperforations of the TM exposing scleral tissue accompanied by blood reflux from the aqueous plexus. The initial ablation zones measured 154 +/- 36 microns in depth and 45 +/- 6 microns in width. Collateral thermal damage zones were 22 +/- 8 microns. At two days post-operative, ablation craters were still blood- and fibrin-filled. The inner surface of the craters were covered with granulocytes. No cellular infiltration of the collateral thermal damage zone was observed. At 10 days post-operative, progressive fibroblastic proliferation was observed, resulting in dense scar tissue formation with anterior synechiae, proliferating capillaries and loss of intertrabecular spaces inside the range of former laser treatment at 60 days post-operative. Trabecular microperforations were closed 60 days after laser treatment in all rabbits. IOP in treated and contralateral eyes did not significantly change its level during whole period of observation. Low-thermal infrared laser energy with minimal thermal damage to collateral structures could not effectively prevent early scarring of trabecular surgery in rabbits.     

Citrobacter sedlakii meningitis and brain abscess in a premature infant

Citrobacter sedlakii was isolated from blood and cerebrospinal fluid cultures of a 5-day-old premature infant with sepsis, meningitis, and brain abscess. This newly described organism was difficult to identify due to discrepancies between the Vitek and API 20E identification systems. To our knowledge, this is the first report of the isolation of C. sedlakii from cerebrospinal fluid.     

Secretory IgA inhibits Pseudomonas aeruginosa binding to cornea and protects against keratitis

PURPOSE: To determine whether secretory IgA (SIgA) antibody inhibits Pseudomonas aeruginosa binding to cornea in vitro and if boosting SIgA antibody in tears using heat-killed P. aeruginosa as an immunizing antigen is protective in vivo in experimentally induced bacterial keratitis in the mouse. METHODS: SIgA, immunoglobulin-G, immunoglobulin-M, and an undiluted crude human milk preparation were tested in vitro for their ability to inhibit P. aeruginosa binding to the scarified corneas of adult (6 weeks to 6 months of age) mice by topical application of each before similar delivery of the bacterial inoculum. Scanning electron microscopy (scanning EM) was used to quantitate bacterial adherence. In vivo mice were immunized topically with heat-killed P. aeruginosa or sham immunized by application of a similar volume of phosphate-buffered saline (PBS). Tears were collected from both groups of mice and levels of immunoglobulins (Igs) measured by enzyme-linked immunosorbent assay (ELISA). After the second immunization, the same two groups were challenged ocularly with 5.0 x 10(7) colony forming units P. aeruginosa and the response to infection graded. RESULTS: In vitro, after a 30-minute preincubation with Igs, SIgA (250 micrograms/ml) significantly decreased P. aeruginosa binding to cornea in vitro when compared to the number of bacteria bound in PBS control specimens, and binding reduction was concentration dependent. In vivo, 15 days after a second ocular topical immunization, tear SIgA was elevated significantly and was specific for P. aeruginosa when measured by ELISA. In vivo, corneal disease response grades in the heat-killed antigen immunized mice also were significantly less severe when compared to sham-immunized mice. CONCLUSIONS: SIgA significantly inhibits binding of P. aeruginosa to the wounded mouse cornea in vitro, and inhibition is concentration dependent. In vivo, specific antipseudomonal SIgA in mouse tears can be elicited by topical ocular immunization with heat-killed P. aeruginosa, and a significant number of immunized animals with elevated levels of SIgA in their tears exhibited less severe ocular disease after bacterial challenge.     

Congenital cholesteatomas in the tympanic membrane

The etiology of congenital middle ear (ME) cholesteatomas is unclear. One etiologic possibility of ME cholesteatoma may be progression of a congenital tympanic membrane (TM) cholesteatoma. We recently have encountered three cases of congenital tympanic membrane cholesteatoma. Each child, ages 1, 3, and 14 years, presented with cholesteatoma of the tympanic membrane extending into the middle ear. These children have not had previous otologic surgery including myringotomy, nor had they had repeated middle ear infections, perforation, or trauma. Neither the 3-year-old nor 14-year-old child complained of hearing loss. Audiograms demonstrated only a mild conductive loss. Each child underwent excision with tympanoplasty. Although the middle ear component of the cholesteatoma was always more extensive than the pearl seen, the point of attachment was the TM and not the middle ear. This demonstrates one possible source for congenital cholesteatomas.     

Differential effects of DNA supercoiling on radical-mediated DNA strand breaks

Supercoiling is an important feature of DNA physiology in vivo. Given the possibility that the reaction of genotoxic molecules with DNA is affected by the alterations in DNA structure and dynamics that accompany superhelical tension, we have investigated the effect of torsional tension on DNA damage produced by five oxidizing agents: gamma-radiation, peroxynitrite, Fe2+/ EDTA/H2O2, Fe2+/H2O2, and Cu2+/H2O2. With positively supercoiled plasmid DNA prepared by a recently developed technique, we compared the quantity of strand breaks produced by the five agents in negatively and positively supercoiled pUC19. It was observed that strand breaks produced by gamma-radiation, peroxynitrite, and Fe2+/EDTA/H2O2 were insensitive to DNA superhelical tension. These results are consistent with a model in which chemicals that generate highly reactive intermediates (e.g., hydroxyl radical), but do not interact directly with DNA, will be relatively insensitive to the changes in DNA structure and dynamics caused by superhelical tension. In the case of Fe2+ and Cu2+, metals that bind to DNA, only Cu2+/H2O2 proved to be sensitive to DNA superhelical tension. Strand breaks produced by Cu2+/H2O2 in the positively supercoiled substrate occurred at lower Cu concentrations than in negatively supercoiled DNA. Furthermore, a sigmoidal Cu2+/H2O2 damage response was observed in the negatively supercoiled substrate but not in positively supercoiled DNA. The results with Cu2+ suggest that the redox activity, DNA binding orientation, or DNA binding affinity of Cu1+ or Cu2+ is sensitive to superhelical tension, while the results with the other oxidizing agents warrant further investigation into the role of supercoiling in base damage.     

Developmental pathways leading to externalizing behaviors in 5 year olds born before 29 weeks of gestation

Recent studies have improved our understanding of the cytogenesis, biology, and therapy of chronic lymphocytic leukemia (CLL). This review highlights this recent progress reported over the past year. We have improved our understanding of the cytogenetic abnormalities in CLL and soon may see identification of new tumor suppressor genes that may be deleted in the leukemia cells of a large number of patients with this disease. We have achieved a better understanding of the surface antigens that help govern the pattern of tissue-infiltration of leukemia cells in vivo. Studies on the immune pathophysiology of CLL are providing clues to potential mechanisms leading to the immunodeficiency associated with this disease. Combination chemotherapy with purine analogues is showing promise for improved efficacy in CLL. Finally, new therapies incorporating bone marrow transplantation, and possibly gene therapy, increasingly are being considered for the therapy of patients with this disease.     

delta-Aminolevulinate dehydratase inhibition by 2,3-dimercaptopropanol is mediated by chelation of zinc from a site involved in maintaining cysteinyl residues in a reduced state

Phospholipase A2 (PLA2) hydrolyses membrane phospholipids (PL) and it may release arachidonic acid (AA)--the precursor of eicosanoids--from the sn-2 position. PLA2 and metabolites of its catalytic activity participate in many processes in the organism: metabolism of lipids, inflammation and immune reactions, membrane and tissue reparation, proliferation, and others. PLA2 as also an important element in the signal transduction. In the present article, PLA2 is characterised, classified into several types, and its mechanism of action together with its possible role in the disease processes are described. The attention is aimed at two forms of PLA2: The secretory (PLA2) of the type II which is associated with the inflammation injury, and the cytosolic PLA2 which is the main catalyst in the liberation of AA and which participates in the signal transduction. Other forms of PLA2 has been also described.