In vitro inhibition of matrix metalloproteinase activity in tracheal epithelial lining fluid from horses with recurrent airway obstruction

OBJECTIVE: To evaluate inhibitory effects of synthetic matrix metalloproteinase (MMP) inhibitors in vitro on gelatinolytic and collagenolytic activities in tracheal epithelial lining fluid (TELF) of horses with recurrent airway obstruction (RAO). ANIMALS: 10 horses with RAO and 5 healthy control horses. PROCEDURES: Substrate-based functional assays, collagen I and gelatin degradation, were used to measure endogenous collagenolytic and gelatinolytic activities in TELF. In vitro inhibition of MMP activity in TELF with 2 chemically modified tetracyclines (CMTs; CMT-3 and CMT-8) and 2 bisphosphonates (BPs; zoledronate and pamidronate) was evaluated. RESULTS: CMT-3, CMT-8, zoledronate, and pamidronate in a dose-dependent manner inhibited TELF type I collagenolytic and gelatinolytic activities, although no complete inhibition of TELF type I collagenolytic and gelatinolytic activities was achieved with the inhibitor concentrations of 25 to 500 microM tested. The CMTs inhibited pathologically induced collagen I degradation more effectively than BPs. Of the tested CMTs, CMT-3 was the most effective inhibitor of gelatinolytic activity, and the efficiency of CMT-3 corresponded with that of the BPs. CONCLUSIONS AND CLINICAL RELEVANCE: An increase in MMP activity in the equine respiratory tract may potentially be inhibited by administration of CMTs or BPs. Distinct synthetic MMP inhibitors may eventually provide an additional means for pharmacologic treatment by decreasing ongoing active tissue destructive inflammation associated with chronic lung disease. The MMP inhibitors such as CMTs and BPs that are targeted to solely inhibit a pathologic increase in MMP activities provide the advantage of minimal adverse effects that are characteristics of other excessively potent MMP inhibitors.     

Direct experimental evidence for the contribution of lime to CO2 release from managed peat soil

Liming is a common management practice used to achieve optimum pH for plant growth in agricultural soils. Addition of lime to the soil, however, may cause CO₂ release when the carbonates in lime dissolve in water. Although lime may thereby constitute a significant carbon source, especially under acidic soil conditions, experimental data on the CO₂ release are lacking so far. We conducted a split-plot experiment within a cut-away peatland cultivated with a bioenergy crop (reed canary grass, Phalaris arundinacea L.) with lime and fertilizer treatments to determine effects of lime on the CO₂ emissions from soil and to better understand mechanisms underlying liming effects. Carbon dioxide release was measured over two growing seasons in the field after liming, and complementary laboratory studies were conducted. To differentiate CO₂ derived from lime and biotic respiration the δ¹³C of CO₂ released was determined and the two-pool mixing model was applied. The results showed that lime may contribute significantly to CO₂ release from the soil. In the laboratory, more than 50% of CO₂ release was attributable to lime-carbonates during short-term incubation. Lime-derived CO₂ emissions were much lower in the field, and were only detected during the first (2–4) months after the application. However, a maximum of 12% of monthly CO₂ emissions from the cultivated peatland originated from the lime. Biotic respiration rates were similar in limed and unlimed soils, suggesting that higher pH did not, at least in the short-term, increase carbon losses from cultivated peat soils. Additional fertilization and acidification did not contribute to further CO₂ release from the lime. According to our first estimations about one sixth of the lime applied would be released as CO₂ from the managed peatland, with all lime-derived emissions occurring during the first year of application (equivalent to about 4.6% of the total annual CO₂ losses from the soil in the first year). This suggests that the mass-balance approach as proposed by the IPCC Tier 1 methodology, which assumes that all carbon in lime ends up as CO₂ in the atmosphere, overestimates the emissions from lime. Our study further shows that there is a great risk to overestimate heterotrophic microbial activity in limed soils by measuring the CO₂ release without separating abiotic and biotic CO₂ production.     

Effect of temperature and glassy states on the molecular mobility of solutes in frozen tuna muscle as studied by electron spin resonance spectroscopy with spin probe detection

The mobility of solutes in frozen food systems (tuna muscle, sarcoplasmic protein fraction of tuna muscle, and carbohydrate-water) has been studied using the temperature dependence of the shape of electron spin resonance (ESR) spectra of the spin probe 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL). The spin probe was incorporated into the tuna meat from an aqueous solution of TEMPOL or by contact with a layer of TEMPOL crystals. The melting/freezing of freeze-concentrated solutes in frozen tuna meat was observed to take place over a range of temperatures from -25 to -10 degrees C. Lower temperatures gave ESR powder spectra due to the decreased mobility of the spin probe, and the temperature dependence of the mobility of the spin probe did not show abrupt changes at the glass transition temperatures of the systems. The mobility of nonglass forming solutes is concluded to be decoupled from the glass forming components. Similar behavior was also observed for TEMPOL in frozen, aqueous carbohydrate systems. The temperature dependence of the mobility of TEMPOL in the frozen systems was analyzed using the Arrhenius equation, and the logarithm of the Arrhenius preexponential factor tau(a) was found to be linearly correlated with the activation energy for all of the tuna and carbohydrate samples, indicating a common molecular mechanism for the observed mobility of TEMPOL in all of the systems. The linear correlation also suggests that the observed mobility of TEMPOL in the frozen aqueous systems is dominated by enthalpy-entropy compensation effects, where the mobility of TEMPOL is thermodynamically strongly coupled to the closest surrounding molecules.     

Fruit development in artificially self- and cross-pollinated strawberries (Fragaria × ananassa) and raspberries (Rubus idaeus)

Tunnel cultivation of berries is increasing on a global basis, and suitable management techniques are being studied for growing berry crops in a protected environment. Soil mulching and irrigation are among the techniques that significantly affect plant growth and yield. Tunnel cultivation is often limited to growing single cultivars, and little attention has been paid to the effects of growing several cultivars in the same tunnel. There are indications that crop yield and quality might benefit from cross-pollination. In this study, the effects of artificial self- and cross-pollination were studied in 15 strawberry and 6 red raspberry cultivars grown in a protected environment in a greenhouse. Four strawberry cultivars benefited from cross-pollination and one required it. In the case of raspberries, self- and cross-pollination had no effect on yield and fruit quality. In conclusion, particular berry cultivars are better suited to tunnel cultivation.     

Survey of macroalgal mats in the Gulf of Finland,Baltic Sea

1. The occurrence of macroalgal mats in shallow coastal waters was studied in the Gulf of Finland, Baltic Sea. 2. Altogether, 89 sites were visited and the near‐shore waters of the islands were investigated, mainly by scuba diver, to about 10 m depth. In 26 places, drifting algal mats were recorded. 3. The macroalgal mats occurred in patches of up to 10 m² in area, in a range of depths from 0.5 to 10 m, with the optimum depth being less than 5 m. The coverage of the mat varied between 10–70%, and the thickness of the mat was found to vary between 2 and 30 cm. The most frequently recorded, dominant species was Cladophora glomerata. 4. In some cases, anoxic conditions were observed as black sediment underneath the mat. This phenomenon could be a sign of very severe eutrophication of the Baltic Sea. Copyright © 2001 John Wiley & Sons, Ltd.     

Occurrence and characterization of methicillin-resistant staphylococci from bovine mastitis milk samples in Finland

Background

Methicillin-resistant staphylococci (MRS) are increasingly being isolated in bovine mastitis. The aim of our study was to evaluate the occurrence of MRS in Finnish mastitis milk samples and characterize the MRS isolates using molecular methods.

Results

Methicillin-resistant S. aureus (MRSA) was a rare finding in bovine mastitis in Finland. Only two out of 135 (1.5%) S. aureus isolates were positive for mec genes. One of these carried mecA and was of spa type t172, SCCmec type IV and ST375, and the other harboured mecC, being spa type t3256, and ST130. MRSA ST375 is common among human MRSA isolates in Finland, but this is the first report in the country of bovine mecC MRSA. In coagulase-negative staphylococci (CoNS) originating from bovine mastitis, methicillin resistance was more common. In the two CoNS collections studied, 5.2% (17/324) and 1.8% (2/110) of the isolates were mecA positive. Eighteen of these were methicillin-resistant S. epidermidis (MRSE), which were divided into 6 separate PFGE clusters. One pulsotype was detected in different parts of the country, indicating clonal spread. Most MRSE (13/18) were of SCCmec type IV, one was of type V and four were non-typeable. Comparison with a human staphylococcal database indicated that bovine MRSE strains were not closely related to human MRSE isolates.

Conclusions

The occurrence of MRS, especially MRSA, in bovine mastitis in Finland was low. Most methicillin-resistant bovine CoNS are MRSE, and we found evidence of a bovine MRSE strain that may spread clonally. This is the first report of a Finnish bovine isolate of MRSA_mecC ST130. The study provides a baseline for further MRS monitoring.     

Microbial communities, biomass, and activities in soils as affected by freeze thaw cycles

Two Finnish agricultural soils (peat soil and loamy sand) were exposed to four freeze-thaw cycles (FTC), with a temperature change from −17.3±0.4 °C to +4.1±0.4 °C. Control cores from both soils were kept at constant temperature (+6.6±2.0 °C) without FTCs. Soil N₂O and CO₂ emissions were monitored during soil thawing, and the effects of FTCs on soil microbes were studied. N₂O emissions were extremely low in peat soil, possibly due to low soil water content. Loamy sand had high N₂O emission, with the highest emission after the second FTC. Soil freeze-thaw increased anaerobic respiration in both soil types during the first 3-4 FTCs, and this increase was higher in the peat soil. The microbial community structure and biomass analysed with lipid biomarkers (phospholipid fatty acids, 3- and 2- hydroxy fatty acids) were not affected by freezing-thawing cycles, nor was soil microbial biomass carbon (MIB-C). Molecular analysis of the microbial community structure with temperature gradient gel electrophoresis (TGGE) also showed no changes due the FTCs. These results show that freezing and thawing of boreal soils does not have a strong effect on microbial biomass or community structure.