ALKALINE PHOSPHATASE ACTIVITIES AMONG POPULATIONS OF THE COLONY-FORMING DIAZOTROPHIC CYANOBACTERIUM TRICHODESMIUM SPP. (CYANOBACTERIA) IN THE RED SEA

Alkaline phosphatase activities of the diazotrophic marine cyanobacterium Trichodesmium were studied among natural populations in the northern Red Sea and in laboratory cultures of Trichodesmium sp. strain WH9601. Open-water tuft-shaped colonies of Trichodesmium showed high alkaline phosphatase activities with 2.4–11.7 μmol p-nitrophenylphosphate (PNPP) hydrolyzed·μg chl a ^{− 1}·h ^{− 1}, irrespective of date or origin of the sample. Coastal populations of the Trichodesmium tuft colonies had low alkaline phosphatase activities with 0.2–0.5 μmol PNPP·μg chl a ^{− 1}·h ^{− 1}. An exception was the Trichodesmium fall maximum, when both tuft colonies and the plankton community (<100 μm) had alkaline phosphatase activities of 0.6–7.4 μmol PNPP·μg chl a ^{− 1}·h ^{− 1}. Likewise, the more rare puff and bow-tie colonies of Trichodesmium spp. in coastal waters had elevated alkaline phosphatase activities (0.8–1.6 μmol PNPP·μg chl a ^{− 1}·h ^{− 1}) as compared with tuft colonies coinhabiting the same waters. Intact filaments of tuft-forming Trichodesmium sp. strain WH9601 from phosphate-replete cultures had a base alkaline phosphatase activity of 0.5 μmol PNPP·μg chl a ^{− 1}·h ^{− 1}. This activity underwent a 10-fold increase in phosphate-deplete cultures and in cultures supplied with glycerophosphate as the sole P source. The elevated level of alkaline phosphatase activity was sustained in P-deplete cultures, but it declined in cultures with glycerophosphate. The decline is suggested to result from feedback repression of alkaline phosphatase synthesis by the phosphate generated in the glycerophosphate hydrolysis. The enhanced alkaline phosphatase activities of Trichodesmium spp. populations provide evidence that P stress is an important factor in the ecology of Trichodesmium in the northern Red Sea.     

Differential Response of Lymphocytes and Neutrophils to High Intensity Physical Activity and to Vitamin C Diet Supplementation

We have determined the effects of chronic vitamin C intake on neutrophil and lymphocyte antioxidant defences during the acute phase immune response induced by intense exercise. Blood samples were taken from 16 voluntary athletes in basal conditions, both immediately after and 1 h after a duathlon competition. Sportsmen's nutrient intakes were determined before the competition. After determining the basal plasmatic ascorbate levels, the results were analysed taking into account the vitamin C intake and their plasmatic levels. Two groups were constituted, the vitamin C supplemented group and the control group, with the dietary vitamin C intake as the only statistical difference between groups. The duathlon competition induced a significant neutrophilia, which was higher in the supplemented group. Lymphocyte antioxidant enzyme activities increased after the competition, with a higher increase in SOD activity in the control group than in the supplemented one. The competition decreased neutrophil antioxidant enzyme activities and neutrophil ascorbate concentration. The decrease in the SOD activity in the supplemented group was higher than in the control group. Finally, the duathlon competition increased the expression of MAC-1 neutrophil adhesion molecule in the supplemented group. High vitamin C intake influenced the response of neutrophils and lymphocytes to oxidative stress induced by exercise, increasing the neutrophil activation.     

Global and grain-specific accumulation of glycoside hydrolase family 10 xylanases in transgenic maize (Zea mays)

In planta expression of cell wall degrading enzymes is a promising approach for developing optimized biomass feedstocks that enable low-cost cellulosic biofuels production. Transgenic plants could serve as either an enzyme source for the hydrolysis of pretreated biomass or as the primary biomass feedstock in an autohydrolysis process. In this study, two xylanase genes, Bacillus sp. NG-27 bsx and Clostridium stercorarium xynB, were expressed in maize (Zea mays) under the control of two different promoters. Severe phenotypic effects were associated with xylanase accumulation in maize, including stunted plants and sterile grains. Global expression of these xylanases from the rice ubiquitin 3 promoter (rubi3) resulted in enzyme accumulation of approximately 0.01 mg enzyme per gram dry weight, or approximately 0.1% of total soluble protein (TSP). Grain-specific expression of these enzymes from the rice glutelin 4 promoter (GluB-4) resulted in higher-level accumulation of active enzyme, with BSX and XynB accumulating up to 4.0% TSP and 16.4% TSP, respectively, in shriveled grains from selected T0 plants. These results demonstrate the potential utility of the GluB-4 promoter for biotechnological applications. The phenotypic effects of xylanase expression in maize presented here demonstrate the difficulties of hemicellulase expression in an important crop for cellulosic biofuels production. Potential alternate approaches to achieve xylanase accumulation in planta without the accompanying negative phenotypes are discussed.     

Clinical Audit of COPD Patients Requiring Hospital Admissions in Spain: AUDIPOC Study

BACKGROUNDS: AUDIPOC is a nationwide clinical audit that describes the characteristics, interventions and outcomes of patients admitted to Spanish hospitals because of an exacerbation of chronic obstructive pulmonary disease (ECOPD), assessing the compliance of these parameters with current international guidelines. The present study describes hospital resources, hospital factors related to case recruitment variability, patients' characteristics, and adherence to guidelines. METHODOLOGY/PRINCIPAL FINDINGS: An organisational database was completed by all participant hospitals recording resources and organisation. Over an 8-week period 11,564 consecutive ECOPD admissions to 129 Spanish hospitals covering 70% of the Spanish population were prospectively identified. At hospital discharge, 5,178 patients (45% of eligible) were finally included, and thus constituted the audited population. Audited patients were reassessed 90 days after admission for survival and readmission rates. A wide variability was observed in relation to most variables, hospital adherence to guidelines, and readmissions and death. Median inpatient mortality was 5% (across-hospital range 0-35%). Among discharged patients, 37% required readmission (0-62%) and 6.5% died (0-35%). The overall mortality rate was 11.6% (0-50%). Hospital size and complexity and aspects related to hospital COPD awareness were significantly associated with case recruitment. Clinical management most often complied with diagnosis and treatment recommendations but rarely (<50%) addressed guidance on healthy life-styles. CONCLUSIONS/SIGNIFICANCE: The AUDIPOC study highlights the large across-hospital variability in resources and organization of hospitals, patient characteristics, process of care, and outcomes. The study also identifies resources and organizational characteristics associated with the admission of COPD cases, as well as aspects of daily clinical care amenable to improvement.     

An investigation of the resolution of inflammation (catabasis) in COPD

Background

Chronic Obstructive Pulmonary Disease (COPD) is characterized by an enhanced inflammatory response to smoking that persists despite quitting. The resolution of inflammation (catabasis) is a complex and highly regulated process where tissue resident macrophages play a key role since they phagocytose apoptotic cells (efferocytosis), preventing their secondary necrosis and the spill-over of their pro-inflammatory cytoplasmic content, and release pro-resolution and tissue repair molecules, such as TGFβ, VEGF and HGF. Because inflammation does not resolve in COPD, we hypothesized that catabasis may be abnormal in these patients.

Methods

To explore this hypothesis, we studied lung tissue samples obtained at surgery from 21 COPD patients, 22 smokers with normal spirometry and 13 non-smokers controls. In these samples we used: (1) immunohistochemistry to assess the expression of CD44, CD36, VEGF and TGFβ in lung macrophages; (2) real time PCR to determine HGF, PPARγ, TGFβ, VEGF and MMP-9 gene expression; and, (3) ELISA to quantify lipoxin A4, a lipid mediator of catabasis.

Results

We found that current and former smokers with COPD showed: (1) more inflammation (higher MMP-9 expression); (2) reduced macrophage surface expression of CD44, a key efferocytosis receptor; and, (3) similar levels of TGFβ, VEGF, HGF, PPARγ, and lipoxin A4 than smokers with normal spirometry, despite the presence of inflammation and disease.

Conclusions

These results identify several potential abnormalities of catabasis in patients with COPD.     

Coat Gene Profiles of Several Cat Populations in Cuba,Costa Rica,Colombia, Paraguay,Chile and Argentina,and Possible Genetic Origins of These Cat Populations

Until the present moment, only a scarce number of Latin American domestic cat populations have been studied from a population genetic standpoint. For this reason, the cat populations of La Havana (Cuba), San José (Costa Rica), Bogotá and Ibagué (Colombia), Asunción (Paraguay), Santiago (Chile) and Buenos Aires (Argentina) were sampled for several coat genes. The results obtained were as follows: (1) there was a strong genetic resemblence between several Hispanic American cat populations (especially, those of Buenos Aires, San José and the two Colombian populations studied) and those from South Western United States (California, Texas and Colorado), which adds suport to the suspicion that these populations probably have a common origin; (2) The cat population of Santiago (Chile), contrarily to the other Hispanic American populations studied, showed a strong genetic resemblance with some Anglo North American populations; and (3) The I (long hair) and d(dilution) alleles showed systematic higher frequencies in the Hispanic American populations than those observed in Spain. Although the Hispanic American populations were not identical to the current Spanish populations (with the exception of Asuncion), this historic genetic experiment was very different to that found for the British populations and their overseas colonies.     

A comparison of linear and daily undulating periodized programs with equated volume and intensity for local muscular endurance

The purpose of this study was to compare linear periodization (LP), daily undulating periodization (DUP), and reverse linear periodization (RLP) for gains in local muscular endurance and strength. Sixty subjects (30 men, 30 women) were randomly assigned to LP, DUP, or RLP groups. Maximal repetitions at 50% of the subject's body weight were recorded for leg extensions as a pretest, midtest, and posttest. Training involved 3 sets (leg extensions) 2 days per week. The LP group performed sets of 25 repetition maximum (RM), 20RM, and 15RM changing every 5 weeks. The RLP group progressed in reverse order (15RM, 20RM, 25RM), changing every 5 weeks. The DUP group adjusted training variables between each workout (25RM, 20RM, 15RM repeated for the 15 weeks). Volume and intensity were equated for each training program. No significant differences were measured in endurance gains between groups (RLP = 73%, LP = 56%, DUP = 55%; p = 0.58). But effect sizes (ES) demonstrated that the RLP treatment (ES = 0.27) was more effective than the LP treatment (control) and the DUP treatment (ES = -0.02) at increasing muscular endurance. Therefore, it was concluded that making gradual increases in volume and gradual decreases in intensity was the most effective program for increasing muscular endurance.     

Nitric oxide synthase type I (nNOS), vascular endothelial growth factor (VEGF) and myoglobin-like expression in skeletal muscle of Antarctic icefishes (Notothenioidei: Channichthyidae)

The Antarctic icefishes Channichthyidae lack haemoglobin and are thought to lack myoglobin (Mb) in their skeletal muscle as well. Due to the absence of both respiratory pigments, icefishes may present a variety of physiological adaptations in their skeletal muscles. In mammals, molecular responses to limiting oxygen availability in the skeletal muscle include, among others, the over expression of nitric oxide synthases (NOS), such as type I (neuronal nNOS) and type III (endothelial eNOS), as well as the vascular endothelial growth factor (VEGF). In this paper, we evaluated by western blot analysis whether the skeletal muscle of haemoglobin-less icefishes expresses in a constitutive manner higher levels of the type I and type III NOS isoforms and VEGF. Our results demonstrate that haemoglobin-less icefish of the family Channichthyidae do indeed present higher expression of the type I NOS isoform compared with red-blooded Antarctic fish species of other families of the same suborder Notothenioidei. In contrast, VEGF was not over-expressed. Moreover, we show that some icefish species, thought previously to lack Mb in oxidative muscles, actually present Mb-like immunoreactivity in their skeletal muscle.     

Golgi enzymes are enriched in perforated zones of golgi cisternae but are depleted in COPI vesicles

In the most widely accepted version of the cisternal maturation/progression model of intra-Golgi transport, the polarity of the Golgi complex is maintained by retrograde transport of Golgi enzymes in COPI-coated vesicles. By analyzing enzyme localization in relation to the three-dimensional ultrastructure of the Golgi complex, we now observe that Golgi enzymes are depleted in COPI-coated buds and 50- to 60-nm COPI-dependent vesicles in a variety of different cell types. Instead, we find that Golgi enzymes are concentrated in the perforated zones of cisternal rims both in vivo and in a cell-free system. This lateral segregation of Golgi enzymes is detectable in some stacks during steady-state transport, but it was significantly prominent after blocking endoplasmic reticulum-to-Golgi transport. Delivery of transport carriers to the Golgi after the release of a transport block leads to a diminution in Golgi enzyme concentrations in perforated zones of cisternae. The exclusion of Golgi enzymes from COPI vesicles and their transport-dependent accumulation in perforated zones argues against the current vesicle-mediated version of the cisternal maturation/progression model.     

Barley hordothionin accumulates in transgenic oat seeds and purified protein retains anti-fungal properties in vitro

Summary Transgenic anti-fungal gene expression in heterologous species provides a means to test resistance protein combinations across species barriers. This is the first report of transgenic anti-fungal seed storage protein accumulation in oat seed. An anti-fungal barley (Hordeum vulgare L.) hordothionin (Hthl) gene was genetically engineered into oat (Avena sativa L.) to determine the effect of hordothionin on pathogen resistance. The transgene was expressed in both leaf and seed tissue, with transgenic protein accumulation occurring only in the seed. Transgenic oat line HTH-Av5 expressed c. 94 μg HTH/g seed, 19% of native barley seed levels. The anti-fungal activities of HTH fractions from barley cv. Morex and oat (transgenic and control) were tested in an in vitro growth assay against an important small grain pathogen. Fusarium graminearum. The partially purified HTH fractions from control oat seeds did not inhibit fungal growth, while HPLC-purified HTH positive control, as well as partially purified barley and transgenic oat HTH inhibited growth similarly over a range of concentrations. These results indicate hordothionin can be expressed in a heterologous cereal species and still maintain its anti-fungal properties. Future studies with HTH targeted to additional tissues are planned to test for increased fungal resistance. The University of Wisconsin and the USDA neither guarantee nor warrant the standard of the products named herein, and the use of the name by University of Wisconsin or USDA implies no approval of the product to the exclusion of others that may also be suitable.