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Animal models for visual deprivation-induced strabismus and nystagmus.

The development of gaze-stabilizing systems depends on normal vision during infancy. Monkeys reared with binocular lid suture (BLS) for the first 25-40 days of life have strabismus, optokinetic nystagmus deficits, latent nystagmus, and decreased binocular cells in the visual cortex and nucleus of the optic tract. When BLS is extended to 55 days, pendular and congenital nystagmus also occurs. Eyelids in infant monkeys are hairless and thin, but BLS still degrades sensory fusion, motion, and form perception. To determine to what extent these visual properties are critical in the development of normal gaze stabilization, we examined infant monkeys reared with one opaque contact lens over one eye, alternated to the fellow eye every other day (AMO); and monkeys reared in a 3-Hz strobe environment. Monkeys reared with AMO develop strabismus, but have normal optokinetic nystagmus and no spontaneous nystagmus. Area 17 is monocular, but the medial temporal area and the nucleus of the optic tract are binocular. Monkeys reared in strobe light develop pendular nystagmus but not strabismus. We were puzzled by the results of the AMO monkeys until we examined infant monkeys with BLS that were prevented from seeing form through the lids. This was done by leaving the tarsal plate intact behind the eyelid. They developed similar to the AMO monkeys. These results suggest that disruption of sensory fusion during infancy (BLS, AMO) causes strabismus. If strabismus occurs while the monkeys have some form vision from each eye (BLS without tarsal plate), then the nucleus of the optic tract becomes monocular, which causes optokinetic nystagmus deficits and latent nystagmus. Infant monkeys reared without visual motion develop pendular nystagmus.

Animals↗

Bacteriocin-like substance inhibits potato soft rot caused by Erwinia carotovora.

Soft rot is a major problem encountered in potatoes during postharvest storage. The soft rot bacterium Erwinia carotovora was inhibited by a novel bacteriocin-like substance (BLS) produced by Bacillus licheniformis P40. The BLS caused a bactericidal effect on E. carotovora cells at 30 microg mL(-1). Transmission electron microscopy showed that BLS-treated cells presented wrinkled bacterial surfaces and shrinkage of the whole cell, indicating plasmolysis. Erwinia carotovora cells treated with BLS were analyzed by FTIR showing differences in the 1390 cm(-1) and 1250-1220 cm(-1) bands, corresponding to assignments of membrane lipids. BLS was effective in preventing E. carotovora spoilage on potato tubers, reducing the symptoms of soft rot at 240 microg mL(-1) and higher concentrations. Soft rot development was completely blocked at 3.7 mg mL(-1). This BLS showed potential to protect potato tubers during storage.

Bacillus↗

Deficient antigen-presenting cell function in multiple genetic complementation groups of type II bare lymphocyte syndrome.

The absence of HLA class II gene expression in type II bare lymphocyte syndrome (BLS) results from defective transcriptional activation of class II histocompatibility genes. Genetic studies have revealed that distinct defects in multiple trans-acting factors result in the immunodeficient BLS phenotype. We studied antigen-presenting cell (APC) function in DR-transfected BLS cells derived from multiple complementation groups. Each BLS cell line displayed the same defective APC phenotype: an inability to mediate class II-restricted presentation of exogenous protein antigens, and structurally altered class II alpha beta dimers. Expression of the HLA class II-like genes DMA and DMB, previously implicated in antigen presentation, was reduced or absent in the BLS cells. Fusion of BLS cells with cell line 721.174, which has a genomic deletion of HLA class II genes, coordinately restores class II structural gene and DM gene expression and a wild-type APC phenotype. Thus each of the molecular defects that silences class II structural gene transcription also results in a defective APC phenotype, providing strong evidence for coregulation of these two functionally linked pathways.

Amino Acid Sequence↗

Cardiopulmonary resuscitation: basic life support skills: 1.

Basic life support (BLS) is potentially life-saving and unnecessary delays in starting BLS must be avoided. Sudden cardiac death occurs within 1 hour of symptoms in 40% of patients, and in 60% of young/middle-aged patients. BLS can support the casualty and save lives. Bystanders, including nurse aiders, should summon professional help before commencing BLS. Although technical expertise and equipment are necessary in specific situations, BLS 'buys time' while waiting for the equipment to arrive. Seventy per cent of cardiac arrests occur in the home and as BLS skills are often the only skills immediately available, their acquisition is essential.

Clinical Competence↗

A polymeric bacterial protein activates dendritic cells via TLR4.

The enzyme lumazine synthase from Brucella spp. (BLS) is a highly immunogenic protein that folds as a stable dimer of pentamers. It is possible to insert foreign peptides and proteins at the 10 N terminus of BLS without disrupting its general folding, and these chimeras are very efficient to elicit systemic and oral immunity without adjuvants. In this study, we show that BLS stimulates bone marrow dendritic cells from mice in vitro to up-regulate the levels of costimulatory molecules (CD40, CD80, and CD86) and major histocompatibility class II Ag. Furthermore, the mRNA levels of several chemokines are increased, and proinflammatory cytokine secretion is induced upon exposure to BLS. In vivo, BLS increases the number of dendritic cells and their expression of CD62L in the draining lymph node. All of the observed effects are dependent on TLR4, and clearly independent of LPS contamination. The described characteristics of BLS make this protein an excellent candidate for vaccine development.

Animals↗

[Educational packages: adult education about basic and advanced methods of life support measures. Analysis of some educational systems].

Educational courses are increasingly being offered in Italy to health care workers who want to acquire or improve their basic (BLS) or advanced (ACLS) resuscitation skills. Since 1994 the Italian Resuscitation Council planned and implemented educational BLS and ACLS packages. Their content, methodology and implementation strategies was analysed on the basis of the educational and informational materials offered to the learners and instructors. The duration of the courses varies (never exceeding the three days): the performance of the participants is evaluated and certified. The courses are open to critical care doctors, nurses, technicians and in some cases to the lay people. They are based on the adult learning principles and the learning strategies are both formal and interactive. The course instructors are doctors and nurses specifically trained after a successful result (with optimal performance) of a BLS and ACLS course. At least 50,000 people attended to a BLS course and a lower number (1270) to an ACLS course. To date, the number of recognised BLS instructors is 229 and 79 for the ACLS courses. The recommended costs are around 120 US $ for a BLS course and 400 US $ for an ACLS course. Although the debate on the effectiveness of such a training on the survival of people treated for a cardiac arrest is still ongoing, the learning packages show the feasibility of a multi-professional approach to a systematic training in a critical health area.

Adult↗

Outcome of cardiac arrests in a Portuguese hospital--evaluation of a hospital cardiopulmonar resuscitation program at one year.

OBJECTIVE: Evaluation of a hospital-wide resuscitation program at one year. METHODS: All records of cardiac arrest calls were collected, logged in a database by the same operator, and analyzed. The cardiac arrest teams consist of a physician and a nurse with ALS (Advanced Life Support) training. Each team has a radio that is activated by a single emergency number. Only cardiac arrest calls were analyzed. RESULTS: Between March 1999 and March 2000 there were 173 emergency team calls. Of these, 120 were cardiac arrest calls (90 in-hospital and 30 from the emergency room--out-of-hospital cardiac arrests). Of the 90 in-hospital cardiac arrests, 61% were male, and median age was 73 years. In 90% of the calls, basic life support (BLS) was started before the arrival of the cardiac arrest team. The immediate cause was cardiac in 39% of the patients. Initial rhythm was ventricular fibrillation in 8%, asystole in 60% and other rhythms in 24% of the patients. Thirty percent presented return of spontaneous circulation (ROSC). There were no differences between those in whom BLS was started before the arrival of the cardiac arrest team and those in whom BLS had not been started. Ten patients (11%) were discharged from hospital. Of the 30 out-of-hospital cardiac arrests, 70% were male, and median age was 69 years. In 97% BLS was started before the arrival of the cardiac arrest team. The immediate cause was cardiac in 30% of patients. Initial rhythm was ventricular fibrillation in 10%, asystole in 73% and other rhythms in 17% of the patients. ROSC was achieved in 27% of the patients. Three patients (10%) were discharged from hospital. There were no differences either in ROSC or in survival to hospital discharge between in-hospital and out-of-hospital cardiac arrests. The state of health previous to cardiac arrest was significantly different between in- and out-of-hospital cardiac arrests: 3% versus 32% healthy, with no functional limitation. The authors conclude that: first, the current records do not enable all the desired goals of the "Utstein style" to be achieved and need to be reviewed; second, 90 to 97% of BLS previous to the arrival of the cardiac arrest team is a good indication of the efficiency of the hospital-wide program, which included training in BLS for all the hospital staff; third, the survival rate, although in accordance with much of the literature, could be improved.

Adolescent↗

Interleukin-6 production in high-grade B lymphomas: correlation with the presence of malignant immunoblasts in acquired immunodeficiency syndrome and in human immunodeficiency virus-seronegative patients.

The mechanisms leading to malignant cell proliferation may differ between the different histologic forms of high-grade non-Hodgkin's lymphomas. To analyze the potential role of interleukin-6 (IL-6) as a growth factor for lymphomatous cells in these different forms, the in situ production of this cytokine was analyzed in lymphomatous samples taken from 24 patients, 18 of whom were human immunodeficiency virus (HIV) infected. Eleven Burkitt's lymphomas (BLs), seven diffuse large-cell lymphomas, and six immunoblastic lymphomas were studied. In situ hybridization experiments showed that the IL-6 gene was expressed in all tissues. The number of IL-6 gene-expressing cells was 7 times higher in the non-BLs than in the BLs, and it was 17 times higher than that of 14 control lymph nodes displaying a benign follicular hyperplasia. Analysis of individual cases indicated that the level of IL-6 gene expression was strongly correlated with the presence of immunoblasts within the malignant clone. In contrast, this level was not correlated with the presence of Epstein-Barr virus genome in the lymphoma or with the HIV status of patients. Immunohistochemical studies with an anti-IL-6 monoclonal antibody showed that IL-6 was produced in non-BLs, but not in BLs. In the former, IL-6 mainly originated from reactive, nonmalignant cells. Immunohistochemical analyses of non-BLs also showed that malignant cells produced the 80-Kd chain of the IL-6 receptor. Taken together, these results suggest that IL-6 may act as a growth factor in some forms of high-grade B lymphomas. The presence of immunoblasts may be an indicator of such forms.

Acquired Immunodeficiency Syndrome↗

Mutant p53 detected in a majority of Burkitt lymphoma cell lines by monoclonal antibody PAb240.

The status of the p53 gene in lymphoblastoid cell lines (LCLs) and Burkitt lymphoma cell lines (BLs) was investigated. Southern blot analysis demonstrated that no major deletions or rearrangements had occurred in the p53 gene in any of the cell lines. The p53 protein was examined by immunoprecipitation using two monoclonal anti-p53 antibodies. PAb1801 recognizes both wild-type and mutant p53. PAb240 reacts exclusively with mutant p53. Fourteen LCLs reacted with PAb1801, but not with PAb240, suggesting that none of them expressed mutant p53. However, one LCL had mutant p53. This LCL differs from other LCLs in that it grows to higher cell densities and has a higher agarose clonability. All BLs expressed p53. Out of 15 BLs, nine (60%) carried mutant p53, as indicated by their reactivity with PAb240. Among the nine BLs with mutant p53, eight Epstein-Barr virus (EBV)-positive. Three out of the six BLs with wild-type p53 were EBV-positive. Multiple EBV-converted sublines all exhibited the same p53 status as the parental line. Our results indicate that the p53 gene is mutated in a majority of Burkitt lymphoma cell lines (BLs), and suggest that p53 mutation contributes to the malignant phenotype of these cell lines.

Antibodies, Monoclonal↗

Blade and balloon atrial septostomy for creation of an adequate interatrial communication.

During a 7 year period from 1984 to 1991, 100 infants underwent either balloon atrial septostomy (BAS) (n = 92) or blade septostomy (BLS) (n = 8). The indication was complete transposition of the great arteries in all the patients. The mean age in the BAS group was 1.8 +/- 1.5 months while that in the infants requiring BLS due to a thick septum was 3.03 +/- 2.29 months. The degree of improvement in arterial oxygen saturation in both groups was satisfactory -27.16 +/- 14.06% in the BAS group vs 23.5 +/- 12.18% in the BLS group. There were no procedure related deaths in the BLS group, through only monoplane fluoroscopy was used in 6/8 patients. Three patients however died following balloon septostomy. We conclude that BLS is a safe and effective alternative to surgical septostomy when performed with due care. Two dimensional echocardiography during BAS enhances the speed and safety of the procedure and helps to identify patients who may require BLS due to a thick interatrial septum.

Cardiac Surgical Procedures↗

Construction injury rates may exceed national estimates: evidence from the construction of Denver International Airport.

BACKGROUND: Construction of Denver International Airport (DIA) provided a unique opportunity to describe the magnitude of injury on a major construction project for which complete data on injury and hours at risk were available for over 32,000 employees working 31 million hours. METHODS: Comprehensive payroll data for all workers, who were paid standard Davis-Bacon wages, allowed calculation of person-hours at risk by job classification. Complete reporting, facilitated by a single workers' compensation plan covering all contracts and by an on-site medical clinic and designated provider system, allowed us to determine both total and lost-work-time (LWT) injury rate per 200,000 hours at risk by industrial sector, company size, and year of construction. Workers' compensation payment rates were calculated and compared with expected loss rates, derived by the National Council on Compensation Insurance, by sector, company size, and year. RESULTS: DIA's overall total injury rates were over twice those published by the Bureau of Labor Statistics (BLS) for the construction industry for each year of DIA construction. Differences in LWT injury rates were more modest. Total injury rates were also at least twice BLS's rates for all contractor sizes. The injury rate pattern by company size at DIA differed from BLS's in that small firms had injury rates that were lower than or comparable to most other size categories; BLS's rates for small firms were lower than those for all but the very largest (250 or more employees) contractors. DIA's total workers' compensation (WC) payment rate of $7.06 per $100 payroll was only 11% higher than Colorado-specific expected loss rates reported by the National Council on Compensation Insurance. DISCUSSION: Complete reporting, facilitated by the existence of a single WC plan, an on-sites medical clinic, and designated medical providers, yielded injury rates significantly higher than previously reported. The relatively small difference between DIA payment rates significantly higher than previously that the discrepancy between DIA's injury rates and national estimates is due to underreporting of non-LWT injuries of the BLS. The burden of on-site work-related construction injury may be higher and more costly than has been evident from national data.

Accidents, Occupational↗

Engineering of a polymeric bacterial protein as a scaffold for the multiple display of peptides.

Protein assemblies with a high degree of repetitiveness and organization are known to induce strong immune responses. For that reason they have been postulated for the design of subunit vaccines by means of protein engineering. The enzyme lumazine synthase from Brucella spp. (BLS) is highly immunogenic, presumably owing to its homodecameric arrangement and remarkable thermodynamic stability. Structural analysis has shown that it is possible to insert foreign peptides at the ten amino terminus of BLS without disrupting its general folding. These peptides would be displayed to the immune system in a highly symmetric three-dimensional array. In the present work, BLS has been used as a protein carrier of foreign peptides. We have established a modular system to produce chimeric proteins decorated with ten copies of a desired peptide as long as 27 residues and have shown that their folding and stability is similar to that of the wild-type protein. The knowledge about the mechanisms of dissociation and unfolding of BLS allowed the engineering of polyvalent chimeras displaying different predefined peptides on the same molecular scaffold. Moreover, the reassembly of mixtures of chimeras at different steps of the unfolding process was used to control the stoichiometry and spatial arrangement for the simultaneous display of different peptides on BLS. This strategy would be useful for vaccine development and other biomedical applications.

Amino Acid Sequence↗

[Differences in nutrient intake using different nutrient databases-- an example].

Mean dietary intake calculated from 25 7-day-food records by means of the three nutrient data bases modified Souci/Fachmann/Kraut (mSFK) 1986/87, extract of Bundeslebensmittelschlüssel (BLS) version 2.1, and extract of BLS version 2.2 revealed comparable results only for four of 27 nutrients considered. The greatest deviations were found for zink, fluoride, iodine, vitamin D, vitamin C, and dietary fiber. Comparing the revised BLS version 2.2 and mSFK, the differences in fluoride, iodine and dietary fiber intake data were markedly lower than found with the comparison of BLS 2.1 and mSFK; statistically significant differences no longer existed for the vitamins C and A (equivalents). As expected, using the mSFK data base with some missing fields for analytical data underestimation of nutrient intake could be shown for the trace elements zink, fluoride and iodine. With regard to the given results of the investigated group, care has to be taken with some nutrient intake data gathered by means of BLS 2.1, too.

Diet Records↗

Basal lamina-associated heparan sulphate proteoglycan in the rat PNS: characterization and localization using monoclonal antibodies.

Cultured rat Schwann cells produce a basal lamina (BL)-associated heparan sulphate proteoglycan (HSPG). The HSPG has an apparent molecular weight of greater than 450 kD, is sensitive to both heparinase and heparitinase and contains a core protein of approximately 400 kD. Two independently derived monoclonal antibodies, B3 and C17, recognize this HSPG. Using B3 and C17, we found that this HSPG, or immunologically related material, is present in BLs throughout the body and in a small number of connective tissue sites without a formed BL. In the PNS it is present in BLs of Schwann cell-axon units, in synaptic and extrasynaptic portions of muscle fibre BL, and in the BLs of satellite cells that ensheath neurons in sympathetic and sensory ganglia. This HSPG is not detectable in the neuropil of the brain and spinal cord. Neurons, Schwann cells and fibroblasts cultured alone do not assemble a BL or accumulate immunocytochemically detectable amounts of this HSPG, but it is present in BLs assembled in myotube and in Schwann cell-neuron cultures. Thus, this HSPG is a component of most, if not all, BLs in the PNS.

Animals↗

Towards early defibrillation--a nurse training programme in the use of automated external defibrillators.

It is essential that all health care professionals are regularly trained in the practice of basic life support (BLS). In most cases of cardiac arrest, the chances of a favourable outcome depend not only on efficient BLS, but also on the early use of defibrillation. In a hospital environment, the first responders are most likely to be members of the nursing staff. The potential advantage of these personnel being able to perform early defibrillation, as well as BLS, is considerable. We describe an initiative in which instruction in the use of automated external defibrillators (AEDs) was introduced into routine BLS training sessions, with the aim of developing this capability. Using specifically-acquired training aids, 43 nurses and nine other members of the non-medical staff were trained in 12 teaching sessions over a period of 4 months. Refresher training was carried out after 6 months and 37 nurses were retrained during six sessions using a similar syllabus. The programme was well-received by all students, and a higher than expected level of motivation and competence was achieved. Throughout retraining all nurses appreciated the key importance of early defibrillation. We conclude that, although the initial workload is high, it is entirely feasible to augment BLS training for health care professionals with instruction in the use of AEDs. We recommend that this potentially life-saving tuition programme be widely adopted.

Automation↗

Future directions for resuscitation research. II. External cardiopulmonary resuscitation basic life support.

In sudden cardiac deaths outside hospitals, the present performance of external cardiopulmonary resuscitation-basic life support (CPR-BLS), as a bridge to advanced life support (ALS) attempts for restoration of spontaneous circulation (ROSC), still yields suboptimal results. Therefore, future education research should develop more effective, simpler and quicker ways to enable everyone to acquire the necessary BLS skills. Individualized self-training by lay persons is being revived. Although airway control and direct mouth-to-mouth ventilation skills are difficult to acquire, they must continue to be taught to the lay public and health professionals, primarily for use on relatives and friends where infection risk is not a problem. In children and trauma victims, steps A and B alone may be lifesavers. The best way to ventilate and oxygenate during the initiation of brief external CPR-BLS should be re-evaluated. There is a great difference between animals and humans in the behavior of the airway and thorax during coma, and thus in the need for added positive pressure ventilation. During chest compressions in humans, steps A and B are needed. Details deserve re-evaluation. The low perfusion pressures (borderline blood flows) produced by standard external CPR remain the most serious limitation of this method. In spite of extensive efforts so far, novel laboratory research to remedy this limitation is important for the development of more effective emergency artificial circulation. The results of such studies are greatly influenced by different details in animal models. Active compression-decompression (ACD) external CPR, also called 'push-pull' CPR, with a plunger-type device used by hand or a machine, and intermittent abdominal compression (IAC) external CPR are both promising modifications of standard external CPR. Both need further experimental and clinical clarification. For BLS, developing a more effective purely manual CPR-BLS method for help in rapid ROSC should be given high priority. Portable external CPR machines need improvements. They will serve for bridging ROSC-resistant cases through transport and ALS attempts, primarily by freeing the hands of health professionals for more effective sophisticated ALS measures.

Cardiopulmonary Resuscitation↗

An antigen detection immunoassay for big liver and spleen disease agent.

An enzyme-linked immunosorbent assay (ELISA) able to detect an antigen associated with infection with big liver and spleen (BLS) agent was developed. The assay is a capture ELISA (C-ELISA) procedure using ELISA plates coated with mouse monoclonal antibody (mAb 1H4) for antigen capture and a polyclonal chicken Ig preparation, extracted from yolk of BLS antibody positive birds, for detection of captured BLS antigen. mAb 1H4 was produced to a partially purified antigen preparation from liver suspensions of naturally BLS infected birds. The C-ELISA was evaluated for specificity and sensitivity in naturally and experimentally BLS infected and uninfected broiler and layer breeds of domestic fowl.

Animals↗

Possible precursors to new hair cells, support cells, and Schwann cells in the ear of a post-embryonic fish.

The sources of new hair cells, support cells, and Schwann cells were identified in the statoacoustic end organs of normal post-embryonic fish (Astronotus ocellatus). S-phase cells, defined as cells that take up 3H-thymidine in preparation for mitosis, and their progeny were visualized using autoradiography. Two types of S-phase cells were found: 'embryonic-like neuroepithelial' (NE) cells and 'basally located S-phase' (BLS) cell. The NE cells had elongated nuclei and processes extending basally and apically. Thirty minutes after the thymidine injection labeled NE cell nuclei were found between hair cell nuclei and support cell nuclei. After two and four hours survival some labeled NE nuclei were closer to the lumen, where they divided. One labeled support cell was seen after four hours survival and one labeled hair cell after nine hours survival. Significant numbers of labeled support and hair cells were not seen until 24 h survival. These results led us to identify NE cells as the immediate source of new hair cells and support cells. The BLS cells had small nuclei with little surrounding cytoplasm. Shortly after the thymidine injection BLS cells were found between the hair cell nuclei and the basement membrane, and they underwent mitosis in this position. The BLS cells resemble intra-epithelial Schwann cells, except that they are not curved around axonal profiles. We suggest that the BLS cells are Schwann cell precursors.

Animals↗