Lignan derivatives from the liverwort Bazzania trilobata

Eight lignan derivatives trilobatin D-K, as well as jamesopyrone were isolated from the liverwort Bazzania trilobata. Their structures have been elucidated based on extensive NMR spectral evidence.     

The tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate enhances the proliferative response of Balb/c-3T3 cells to hormonal growth factors.

Stimulation of Balb/c-3T3 cell growth by TPA requires factors found in serum. We examined the interaction between TPA and serum growth factors in the stimulation of cell growth. The number of cells synthesizing DNA (incorporating 3H-thymidine) within 24 to 30 hours after the addition of TPA and the growth factors to density-inhibited Balb/c-3T3 cultures in serum-free medium was determined by autoradiography. With no additions or with TPA (30--300 ng/ml) alone, only 3--7% of cells synthesized DNA. However, TPA synergistically promoted DNA synthesis in combination with each of the defined serum growth fractions, platelet derived growth factor and platelet poor plasma. TPA also synergistically promoted DNA synthesis in combination with purified growth factors including fibroblast growth factor, insulin (10(-6)--10(-5)M), and epidermal growth factor. In all conditions, TPA enhancement of DNA synthesis also resulted in an increase in cell number. Because TPA synergistically enhanced the activity of each growth factor tested, it did not act identically to any of the growth factors.     

The lipid bilayer of acetylcholine receptor clusters of cultured rat myotubes is organized into morphologically distinct domains

We have studied the composition and organization of the lipid bilayer at the large, substrate-associated clusters of acetylcholine receptors (AChR) that form in cultured rat myotubes. These clusters have a characteristic morphology consisting of alternating linear domains of AChR-rich and AChR-poor membrane, the latter involved in attaching the myotube to the substrate. We partially purified AChR clusters by extracting cultured rat myotubes with the cholesterol-specific detergent, saponin. The lipid bilayer of the cluster preparation was analyzed biochemically and the substructure of the bilayers was studied morphologically using the fluorescent probes, dansyl polymyxin B, and 3,3'-di(C12H25 and C18H37) indocarbocyanine iodide (C12- and C18-diI). Our results demonstrate that preparations of AChR clusters have a lipid composition biochemically similar to that of the surrounding plasma membrane. Morphologically, however, the lipid bilayer appears to be arranged into domains that resemble the interdigitating pattern seen for the AChR. This distinctive lipid organization is not due to the use of saponin to purify clusters, as we obtained similar results with clusters isolated by physically shearing myotube cultures. The domain-like organization of the bilayer at clusters is disrupted by treatments that disperse AChR clusters in intact myotubes or that remove peripheral membrane proteins from isolated clusters. This suggests that such proteins may contribute to the organization of the bilayer. Two additional factors may also contribute to the organization of the bilayer: physical constraints imposed by sites of substrate attachment and, to a lesser extent, "boundary" lipid associated with AChR.     

A histological and immunohistochemical study of the effects of N-acetyl cysteine on retinopathy of prematurity by modifying insulin-like growth factor-1

Retinopathy of prematurity (ROP) is a vasoproliferative disorder that occurs in premature infants and may lead to permanent visual impairment. We investigated both the possible protective role of N-acetyl cysteine (NAC) for preventing ROP and the role of IGF-1 in the disorder. Forty-five newborn rats were divided into three groups. Group 1 was raised in room air as controls. Group 2 was exposed to 60% oxygen for 14 days after birth, then transferred to room air. Group 3 was exposed to the same conditions as group 2, but received intraperitoneal injections of NAC on postnatal days 7–17. After 35 days, both eyes of all rats were processed for histology. Some sections were stained with hematoxylin and eosin to assess structural changes and other sections were immunostained to determine the location of IGF-1. Frozen sections also were prepared and stained for adenosine triphosphatase to detect retinal blood vessels. Compared to the controls, more blood vessels, many of which were abnormal, and increased IGF-1 expression were observed in group 2. In group 3, abnormal blood vessels and IGF-1 expression were less evident. NAC appeared to be an effective vascular-protective agent for ROP by decreasing IGF-1 expression.     

Measuring the spermosphere colonizing capacity (spermosphere competence) of bacterial inoculants

Spermosphere establishment by bacteria which were coated onto seeds was studied using soybean seeds treated with four bacterial strains at levels of log10 1 to 4 colony-forming units (cfu) per seed planted in a field soil mix, and incubated 48 h. Each strain at every inoculum level developed spermosphere population densities of log10 4 to 8 cfu/seed, demonstrating an average multiplication of log10 3 cfu/seed. An alternative method was developed to differentially rank bacteria for spermosphere colonizing capacity, based upon incorporation of bacteria into a soil and monitoring the resulting spermosphere population densities around noninoculated seeds after 4 days at 14 degrees C. Fifty-seven bacterial strains which were isolated from soybean roots or from water samples, including Pseudomonas putida, P. putida biovar B, P. fluorescens, Serratia liquefaciens, Enterobacter aerogenes, and Bacillus spp. were tested in the spermosphere colonization assay. Average spermosphere population densities for the 57 strains ranged from 0 to log10 7.0 cfu/seed. Strains of a given taxon demonstrated marked diversity with ranges from 0 to log10 6.0 cfu/seed for Bacillus spp. and from log10 1.4 to 7.0 cfu/seed for Pseudomonas putida. The relative ranking of representative strains was consistent in repeating experiments. The potential usefulness of the assay for efforts to develop competitive bacterial inoculants for crop seeds is discussed.     

Nitrogen-fixing pseudomonads isolated from roots of plants grown in the canadian high arctic

Root-associated bacteria capable of reducing acetylene to ethylene (biological nitrogen fixation) were isolated from various native plants grown in the Canadian High Arctic. All the strains belonged to the genus Pseudomonas but varied in several physiological characteristics. The rates of acetylene reduction at 14 or 20 degrees C were higher than at 25 or 9 degrees C. Six strains reduced acetylene at 4 degrees C. All the strains exhibited chemotaxis to l-asparagine in semisolid agar at 4 to 25 degrees C. Eleven strains colonized roots of canola (Brassica campestris cv. Tobin) in field soil at population densities of log 4.3 to log 5.1 CFU/g of fresh root at 14 degrees C and log 4.0 to log 5.2 CFU/g of fresh root at 25 degrees C. Some of these nitrogen-fixing pseudomonad strains demonstrated a competitive advantage for root colonization over other rhizosphere bacteria at low temperatures. The combined capabilities of nitrogen fixation and root colonization by diazotrophic pseudomonads may be useful for the development of a biofertilizer inoculant for temperate and cold regions.     

Patterns of inflammation and the use of reversibility testing in smokers with airway complaints

Background

Although both smoking and respiratory complaints are very common, tools to improve diagnostic accuracy are scarce in primary care. This study aimed to reveal what inflammatory patterns prevail in clinically established diagnosis groups, and what factors are associated with eosinophilia.

Method

Induced sputum and blood plasma of 59 primary care patients with COPD (n = 17), asthma (n = 11), chronic bronchitis (CB, n = 14) and smokers with no respiratory complaints ('healthy smokers', n = 17) were collected, as well as lung function, smoking history and clinical work-up. Patterns of inflammatory markers per clinical diagnosis and factors associated with eosinophilia were analyzed by multiple regression analyses, the differences expressed in odds ratios (OR) with 95% confidence intervals.

Results

Multivariately, COPD was significantly associated with raised plasma-LBP (OR 1.2 [1.04–1.37]) and sTNF-R55 in sputum (OR 1.01 [1.001–1.01]), while HS expressed significantly lowered plasma-LBP (OR 0.8 [0.72–0.95]). Asthma was characterized by higher sputum eosinophilic counts (OR 1.3 [1.05–1.54]), while CB showed a significantly higher proportion of sputum lymphocytic counts (OR 1.5 [1.12–1.9]). Sputum eosinophilia was significantly associated with reversibility after adjusting for smoking, lung function, age, gender and allergy.

Conclusion

Patterns of inflammatory markers in a panel of blood plasma and sputum cells and mediators were discernable in clinical diagnosis groups of respiratory disease. COPD and so-called healthy smokers showed consistent opposite associations with plasma LBP, while chronic bronchitics showed relatively predominant lymphocytic inflammation compared to other diagnosis groups. Only sputum eosinophilia remained significantly associated with reversibility across the spectrum of respiratory disease in smokers with airway complaints.     

Prediction and course of symptoms and lung function around an exacerbation in chronic obstructive pulmonary disease

Background

Frequent exacerbations induce a high burden to Chronic Obstructive Pulmonary Disease (COPD). We investigated the course of exacerbations in the published COSMIC study that investigated the effects of 1-year withdrawal of fluticasone after a 3-month run-in treatment period with salmeterol/fluticasone in patients with COPD.

Methods

In 373 patients, we evaluated diary cards for symptoms, Peak Expiratory Flow (PEF), and salbutamol use and assessed their course during exacerbations.

Results

There were 492 exacerbations in 224 patients. The level of symptoms of cough, sputum, dyspnea and nocturnal awakening steadily increased from 2 weeks prior to exacerbation, with a sharp rise during the last week. Symptoms of cough, sputum, and dyspnea reverted to baseline values at different rates (after 4, 4, and 7 weeks respectively), whereas symptoms of nocturnal awakening were still increased after eight weeks. The course of symptoms was similar around a first and second exacerbation. Increases in symptoms and salbutamol use and decreases in PEF were associated with a higher risk to develop an exacerbation, but with moderate predictive values, the areas under the receiver operating curves ranging from 0.63 to 0.70.

Conclusions

Exacerbations of COPD are associated with increased symptoms that persist for weeks and the course is very similar between a first and second exacerbation. COPD exacerbations are preceded by increased symptoms and salbutamol use and lower PEF, yet predictive values are too low to warrant daily use in clinical practice.     

Actigraphic motor activity during sleep from infancy to adulthood

A secondary analysis of longitudinal and cohort studies was carried out to quantitatively investigate the motor activity pattern, recorded through actigraphy, during the first six hours of nocturnal sleep. The first study was of longitudinal nature. Ten healthy participants (four females) were monitored three times, at baseline (T1) when they were infants (mean age 7.10 ± 0.32 months), at the first follow-up examination (T2) around 4 months later (mean age 11.20 ± 0.63 months) and at the second follow-up (T3) around three years later, when they were preschoolers (mean age 4.68 ± 0.14 years). At T1, T2 and T3 each participant wore the actigraph Basic Mini-Motionlogger (Ambulatory Monitoring, Inc., Ardsley, NY, USA) over at least two consecutive nycthemeral cycles, with the aim to measure the mean hourly motor activity count. Seven- and 11-month-old infants had a higher level of motor activity over the night compared to preschoolers. Furthermore, motor activity increased as the night progressed, with a pronounced increment at both T1 and T2, while at T3 such an increase was less marked. The second study was cross-sectional and aimed to explore the motor activity pattern, using actigraphy, during the first six hours of nocturnal sleep in multiple-age healthy groups, from infancy to adulthood. We assigned participants to eight groups according to age: 20 (five females) aged around 10 months old (mean age 10.65 ± 0.67 months); 13 (nine females) aged around 4 years (mean age 4.38 ± 0.51 years); 21 (10 females) aged around 10 years (mean age 9.67 ± 0.91 years); 21 (nine females) aged around 20 years (mean age 19.33 ± 2.44 years); 20 (10 females) aged around 30 years (mean age 29.80 ± 1.99 years); 20 (15 females) aged around 40 years (mean age 40.70 ± 1.26 years); 20 (11 females) aged around 50 years (mean age 50.15 ± 2.80 years) and 20 (nine females) aged around 60 years (mean age 59.25 ± 3.23 years). The participants aged between 10 and 60 years wore the actigraph Basic Mini-Motionlogger over seven consecutive nycthemeral cycles (infants and preschoolers wore the actigraph over at least two consecutive nycthemeral cycles), with the aim to measure the mean hourly motor activity count. The results indicated a significantly higher motor activity count in 10-month-old infants compared to all the remaining age groups. Moreover, the pattern of motor activity of 10-month-old infants was different from that of all other groups, with the highest motor activity counts from the second to the sixth hour of sleep. Considered as a whole, the results of both studies converge regarding the high motor activity detected among infants, which could be explained by the presence of a maturational process that has not yet been fully completed at this stage of life. In both studies, only the motor activity of infants was above the cutoff level established for normal adults, highlighting the need to establish a specific cutoff value for infants.