To elucidate the cytokine-producing capacity of alveolar macrophages (AMs), we have introduced a method of flow cytometry combined with saponin treatment to detect the cell-associated cytokines. We ...studied bronchoalveolar lavage fluid cells from six patients with sarcoidosis (SAR) and six control (CTL) subjects. Cells were either left uncultured, or cultured with and without lipopolysaccharide (LPS), then treated with paraformaldehyde and saponin and analysed for cell-associated interleukin-1 beta (IL-1 beta) and tumour necrosis factor-alpha (TNF-alpha) by flow cytometry. Cell-associated IL-1 beta and TNF-alpha were also analysed by immunoassays. The flow cytometric cytokine values were correlated with the immunoreactive cell-associated cytokines (IL-1 beta: r = 0.45, p < 0.05; TNF-alpha: r = 0.82, p < 0.001). The histograms of cell-associated IL-1 beta yielded a single peak both in the patients and controls, whereas the histograms of cell-associated TNF-alpha exhibited two peaks in SAR patients, but just a single peak in the CTL subjects. The mean value of the cell-associated TNF-alpha in LPS+ AMs was higher in the SAR patients than in the CTL subjects (p < 0.001). In conclusion, the flow cytometric method can be applied to the semiquantitative detection of cell-associated cytokines in alveolar macrophages at the single cell level.
The purpose of this study was to determine the effects of professional subgingival irrigation, together with subsequent patient administered home marginal irrigation, on the incidence of bacteremia ...after scaling and root planing (Sc/RP). A total of 60 periodontal maintenance patients were assigned to either Group 1: subgingival irrigation, with 0.12% CHX and daily marginal irrigation with 0.04% CHX; Group 2: subgingival irrigation with 0.12% CHX and daily marginal irrigation with water; Group 3: subgingival and daily marginal irrigation with water; Group 4: Non-irrigation (control). Patients entered the study after receiving a thorough periodontal maintenance appointment including a complete examination, Sc/RP, and standard oral hygiene instruction. Blood samples were taken at the 3-month visit before and after Sc/RP. Microbiological culturing was done using the Septi-Chek system, selective and non-selective media. Results from 54 patients showed that bacteremia was detected prior to Sc/RP in 2 patients and after Sc/RP in 10 patients. No significant effect by treatment regimens on post Sc/RP bacteremia could be detected. The organisms isolated included Eubacterium lentum, Propionibacterium acnes, Streptococcus species, Neisseria species, Candida albicans, Staphylococcus species, and un-identified Gram-negative rods.
The clinical and immunoregulatory effects of long‐term macrolide antibiotic therapy for patients with chronic lower respiratory tract infections (CLRTI) were investigated.
Clinical parameters and ...neutrophil chemotactic mediators in the epithelial lining fluid (ELF) of CLRTI patients (n=10) were examined before and after 3 months oral administration of roxithromycin (RXM). The in vitro effects of RXM were also examined on the release of these mediators from alveolar macrophages (AM) and neutrophils.
Arterial oxygen tension (p<0.05), vital capacity (VC) (p<0.001), %VC (p<0.05) and forced expiratory volume in one second (p<0.01) were improved after RXM treatment, but airway bacteria were not eradicated. Among the mediators, the levels of interleukin (IL)‐8, neutrophil elastase (NE) and leukotriene B4 (LTB4) were higher in ELF than in plasma of CLRTI patients and they decreased after RXM treatment (n=7, p<0.05 for each). RXM concentrations were significantly increased in the bronchoalveolar lavage cells of the treated patients. In in vitro experiments, RXM showed inhibitory effects on IL‐8 release from AM and neutrophils.
In conclusion, interleukin‐8, neutrophil elastase and leukotriene B4 contribute to the neutrophilic inflammation in the airways of chronic lower respiratory tract infection patients and the clinical effects of roxithromycin may, in part, be attributable to the suppression of excess release of the chemotactic mediators from inflammatory cells.
SDZ MRL 953 (SDZ), a novel immunostimulatory lipid A analog, has been reported to have immunopharmacological activities similar to those of lipopolysaccharide (LPS) but to have little of the toxicity ...of LPS. We investigated the effects of pretreatment with SDZ on Escherichia coli endotoxin-induced acute lung injury in guinea pigs. Four experimental groups consisted of saline control (n = 16), SDZ (-12 h) plus LPS (2 mg/kg of SDZ per kg of body weight injected intravenously 12 h before intravenous injection of 2 mg of LPS per kg; n = 15), SDZ (-10 min) plus LPS (SDZ injected 10 min before LPS injection; n = 10), and LPS alone (n = 16). The animals were sacrificed, and lung tissue was sampled 4 h after LPS or saline infusion. Lung injury was assessed by measuring the wet weight-to-dry weight ratio and the level of 125I-labeled albumin accumulation in bronchoalveolar lavage fluid relative to that in plasma. In the SDZ (-12 h) plus LPS group, these two parameters of acute lung injury were decreased compared with those in the LPS alone group. However, they were not decreased in the SDZ (-10 min) plus LPS group. We conclude that SDZ attenuates endotoxin-induced acute lung injury when it is administered 12 h before LPS injection. The attenuating effects of SDZ are speculated to be due to down regulation of the response to endotoxin rather than to receptor blocking.
ONOUE, S., WAKI, Y., NAGANO, Y., SATOH, S., KASHIMOTO, K.
Neuromodulatory Effects of VIP/PACAP on PC-12 Cells Are Associated with Their N-terminal Structures. PEPTIDES xx(xx) 000–000, 200x.– The ...current study explored whether the differences in biological activities in PC-12 cells between vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are attributable to the sequence difference in their N-terminal portions and are correlated with the solution structures of the peptides. In the neurite outgrowth assay, N-terminal modification of VIP to PACAP-like sequences altered its effect, the activity was confirmed even at a low concentration (10
−10 M). On the contrary, N-terminal modification of PACAP 27 to VIP-like sequences reduced its activity. These relationships were also confirmed for the inhibitory effects of the peptide analogues on PC-12 cells growth at 10
−7 M. The present results combined with our previously reported data, including binding assay, support that the N-termini of VIP/PACAP plays an important role in their activities.
In the past, various tracheotomy incisions have been used at the Children's Hospital of Michigan with the occurrence of complications related to accidental decannulation and immediate recannulation. ...Since that time the inferior based tracheal cartilage flap has been used to minimize early complications. A retrospective study of 126 pediatric tracheotomies performed at the Children's Hospital of Michigan from June 1986 to January 1991 was reviewed. Only tracheotomies performed by a staff otolaryngologist utilizing the inferior based tracheal cartilage flaps were reviewed. This study includes patients with a 6 month to 5 year follow up. The early complication rate was 4%, while the late was 50%. Stomal granulation tissue was comparatively increased in this series of patients but did not hinder decannulation. We consider the use of the inferior based tracheal cartilage flap in the pediatric population a safe and effective technique without increasing the morbidity of long-term tracheotomy.
M. Kanazawa, N. Hasegawa, T. Urano, K. Sayama, S. Tasaka, F. Sakamaki, H. Nakamura, Y. Waki, T. Terashima, S. Fujishima and al. et
Department of Medicine, School of Medicine, Keio University, Tokyo, ...Japan.
Estimating blood content in the lung remains a key step in calculating lung
water volume and microvascular permeability. We studied the effect of
regional lung hematocrit (Hct) variation on assessment of acute lung
injury. Escherichia coli endotoxin was administered in guinea pigs
intravenously. Lung injury was evaluated by measuring the wet-to-dry weight
ratio (W/D) and transvascular 125I-labeled albumin leakage for 3 h
tissue-to-plasma 125I-albumin ratio (T/P) in five tissue samples from
each animal. Residual blood content was corrected using either 51Cr-red
blood cells as a blood cell marker, 99mTc-albumin as a plasma marker, or
both, injected 10 min before the guinea pigs were killed. Lung Hct,
estimated from the marker counts of lung and peripheral blood samples, was
lower than peripheral blood Hct; intraindividual variation, represented by
the standard deviation in each subject, was 0.024 +/- 0.015 for the control
group (coefficient of variation 8.0 +/- 5.1%) and 0.026 +/- 0.013 for the
endotoxin group (coefficient of variation 8.5 +/- 4.1%). Uncorrected W/D
for residual blood content was greater than the corrected W/D.
99mTc-albumin correction gave values closer to the W/D corrected by both
markers. T/P corrected by 99mTc-albumin showed smaller data variations than
the values obtained with 51Cr-red blood cell correction, which was affected
by variations in lung Hct. We recommend using a plasma marker to correct
for blood content in assessing acute lung injury by W/D and T/P.
S. Tasaka, A. Ishizaka, T. Urano, K. Sayama, F. Sakamaki, H. Nakamura, T. Terashima, Y. Waki, K. Soejima, S. Fujishima and al. et
Department of Medicine, School of Medicine, Keio University, Tokyo, ...Japan.
Hyperoxic lung injury is attributable to oxygen radicals produced under
hyperoxic conditions. The 21-aminosteroid (AS), U-74389G, is a potent
antioxidant. We examined the effect of U-74389G on lung injury in guinea
pigs during exposure to 90% O2 for 48 h. We injected either vehicle or 10
mg/kg of U-74389G 30 min before the O2 exposure and injected the same dose
12, 24, and 36 h later. We performed two series of experiments after
exposure. In the first series, we measured the clearance rate of
99mTc-labeled dialdehyde starch (DAS) from the lungs as an index of
pulmonary epithelial damage in three experimental groups consisting of 1)
control (n = 6) O2 alone (n = 6), and 3) O2 + AS (n = 6). In the second
series, pulmonary endothelial injury was estimated by using 28 guinea pigs
divided into four experimental groups consisting of 1) control (n = 8), 2)
AS alone (n = 5), 3) O2 alone (n = 6), and 4) O2 + AS (n = 9). In the
second series, we measured the wet-to-dry weight ratio (W/D) as an index of
lung water and the concentration ratio of 125I-labeled albumin in lung
tissue and bronchoalveolar lavage (BAL) fluid compared with plasma (T/P and
BAL/P, respectively) as indexes of pulmonary endothelial damage. Cell
accumulation in BAL fluid and lung tissue samples was also assessed in the
second series.
This paper discusses the stress dependence of magnetizing properties in non-oriented electrical steel sheets (NO). Compression stresses deteriorate the magnetic properties at low flux density, but ...improve them at high flux density, where magnetization rotations are induced. Tensile stresses reduce magnetizing fields at low flux density, but increase them at high flux density. The stress dependence of NO is affected by the exciting direction, as the rolling direction of NO is easy to magnetize and sensitive to stresses. High-grade types of NO (50A290, etc), which have high Si content and large crystalline grains, are more sensitive to stresses than low grades (50A1300, etc), and the magnetizing fields of high grades at high compression are higher than those of low grades. This stress dependence may depend on magnetization behaviors (domain wall displacement and magnetization rotation) and magnetostriction. Domain wall displacement mainly occurs at lower flux density, and in this case tensile stresses improve magnetic properties and compression deteriorate them because magnetic domains orient the magnetization to and λ100 is positive. However, magnetizations rotate to exciting directions at high flux densities, with the result that the component of magnetization is induced, and in this case compression improves the magnetizing properties, because λ111 is negative, whereas tensile stress deteriorates them. Therefore, stress dependences may be modeled by magnetization behaviors and magnetostriction, and analysis of the magnetic properties using this model of stress dependence may improve NO steels and their applications.
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