Digitonin reaction in electron microscopy

Summary The ultrastructural localization of cholesterol (3-hydroxysterols) has been studied by the digitonin reaction adapted to the electron microscope. The reaction was undertaken in the adrenal gland, liver, and seminiferous tubules of adult albino rats. The digitonincholesterol crystals formed in the reaction proved to be an osmiophilous cylindrical structure made up of coaxial lamellae.     

Further studies of urea-catalyzed phosphorylation reactions

Summary We have analyzed the products formed when mixtures of a nucleoside and ammonium dihydrogen phosphate are heated with an excess of urea. If there is more phosphate than nucleoside in the mixture, compounds containing pyrophosphate bonds are obtained. If uridine, as nucleoside, is in excess over phosphate, di- and oligonucleotides are formed.On leave from Department of Medical Biochemistry, University of Göteborg, Sweden.     

Sequence variation in nuclear ribosomal small subunit,internal transcribed spacer and large subunit regions of Rhizophagus irregularis and Gigaspora margarita is high and isolate‐dependent

Arbuscular mycorrhizal (AM) fungi are known to exhibit high intra‐organism genetic variation. However, information about intra‐ vs. interspecific variation among the genes commonly used in diversity surveys is limited. Here, the nuclear small subunit (SSU) rRNA gene, internal transcribed spacer (ITS) region and large subunit (LSU) rRNA gene portions were sequenced from 3 to 5 individual spores from each of two isolates of Rhizophagus irregularis and Gigaspora margarita. A total of 1482 Sanger sequences (0.5 Mb) from 239 clones were obtained, spanning ~4370 bp of the ribosomal operon when concatenated. Intrasporal and intra‐isolate sequence variation was high for all three regions even though variant numbers were not exhausted by sequencing 12–40 clones per isolate. Intra‐isolate nucleotide variation levels followed the expected order of ITS > LSU > SSU, but the values were strongly dependent on isolate identity. Single nucleotide polymorphism (SNP) densities over 4 SNP/kb in the ribosomal operon were detected in all four isolates. Automated operational taxonomic unit picking within the sequence set of known identity overestimated species richness with almost all cut‐off levels, markers and isolates. Average intraspecific sequence similarity values were 99%, 96% and 94% for amplicons in SSU, LSU and ITS, respectively. The suitability of the central part of the SSU as a marker for AM fungal community surveys was further supported by its level of nucleotide variation, which is similar to that of the ITS region; its alignability across the entire phylum; its appropriate length for next‐generation sequencing; and its ease of amplification in single‐step PCR.     

Immunogenicity and protective efficacy of recombinant iron superoxide dismutase protein from Bordetella pertussis in mice models

Whooping cough (pertussis) is a highly contagious respiratory infection caused by Bordetella pertussis. Although availability of effective pertussis vaccines reportedly decreases the incidence of the disease, B. pertussis circulation in populations has not been eliminated. Thus, it is necessary to find new protein candidates with greater immune protective capacities than the currently available acellular pertussis vaccines. In this study, iron superoxide dismutase (FeSOD) gene (sodB) was cloned, expressed in Escherichia coli and recombinant FeSOD protein thence purified. The recombinant protein (rFeSOD) was formulated with aluminum hydroxide (Alum) or monophosphoryl lipid A (MPLA) and injected intraperitoneally to immunize mice, after which IgG1, IgG2a and IFN‐γ titers were measured to assess humoral and cellular responses, respectively, to these immunizations. The extent of bacterial colonization in lungs of intranasally challenged mice was determined 5, 8 and 14 days post‐challenge. IgG1 and IgG2a responses were significantly stronger in mice that had been immunized with rFeSOD–MPLA than in those that had received rFeSOD‐Alum (P < 0.05). Additionally, IgG2a titers were higher in mice vaccinated with recombinant protein FeSOD (rFeSOD) formulated with MPLA, especially after the second immunization. Immunization with rFeSOD–MPLA also provided a modest, but significant decrease in bacterial counts in lungs of mice (P < 0.05). Antigen specific‐IFN‐γ responses were significantly stronger in the group vaccinated with rFeSOD–MPLA, which could account for the lower bacterial counts. These findings suggest that rFeSOD protein formulated with MPLA has potential as an acellular pertussis vaccine candidate component.     

Ameliorative Effect of Carvacrol on Cisplatin‐Induced Reproductive Damage in Male Rats

Cisplatin (CP) treatment causes the damage in male reproductive system. Carvacrol (CARV) is an antioxidant that is naturally found in some plants. We aimed to investigate the effect of CARV on CP‐induced reproductive toxicity in male rats. Eighteen adult male Sprague–Dawley rats were used. The control group (n = 6) was treated orally with physiological saline (PS) daily for 14 days and a single intraperitoneal (IP) PS injection on day 10. The CP group (n = 6) was administered with daily oral PS for 14 days and a single IP injection of 10 mg/kg CP on day 10. The CARV + CP group (n = 6) was treated with daily 75 mg/kg oral CARV for 14 days and a single IP injection of 10 mg/kg CP on day 10. CP treatment caused the damage on some spermatological parameters (motility, live sperm rate, and abnormal sperm rate), increased the oxidative stress, and induced testicular degeneration and apoptosis. However, CARV treatment mitigates CP‐induced reproductive toxicity.     

Isolation and characterization of long-chain alkane–degrading Acinetobacter sp. BT1A from oil-contaminated soil in Diyarbakır,in the Southeast of Turkey

A strain of long-chain alkane–degrading bacteria, BT1A, was isolated from oil-contaminated soil in Diyarbak?r, in the southeast of Turkey. Morphological, biochemical, and physiological characterization and 16S rRNA gene sequence analysis showed that the strain BT1A was a member of Acinetobacter genus, and it was found to be closely related to Acinetobacter baumannii. The strain BT1A was able to utilize crude petroleum as carbon and energy sources in order to grow. Among the aliphatic hydrocarbons, growth was observed only in the medium containing long-chain alkanes (tridecane, pentadecane, and hexadecane) and squalene. Hexadecane was the most preferred hydrocarbon among the long-chain alkanes. Gas chromatography–mass spectrometry (GC-MS) analysis showed that BT1A degraded 83% of n-alkanes of 1% crude oil in 7 days. The present study indicates that the isolated strain can well be used for biodegradation of hydrocarbons in oil-contaminated sites.     

Net Zero Fort Carson: Integrating Energy,Water, and Waste Strategies to Lower the Environmental Impact of a Military Base

Military bases resemble small cities and face similar sustainability challenges. As pilot studies in the U.S. Army Net Zero program, 17 locations are moving to 100% renewable energy, zero depletion of water resources, and/or zero waste to landfill by 2020. Some bases target net zero in a single area, such as water, whereas two bases, including Fort Carson, Colorado, target net zero in all three areas. We investigated sustainability strategies that appear when multiple areas (energy, water, and waste) are integrated. A system dynamics model is used to simulate urban metabolism through Fort Carson's energy, water, and waste systems. Integrated scenarios reduce environmental impact up to 46% from the 2010 baseline, whereas single‐dimension scenarios (energy‐only, water‐only, and waste‐only) reduce impact, at most, 20%. Energy conserving technologies offer mutual gains, reducing annual energy use 18% and water use 15%. Renewable energy sources present trade‐offs: Concentrating solar power could supply 11% of energy demand, but increase water demand 2%. Waste to energy could supply 40% of energy demand and reduce waste to landfill >80%, but increase water demand between 1% and 22% depending on cooling system and waste tonnage. Outcomes depend on how the Fort Carson system is defined, because some components represent multiple net zero areas (food represents waste and energy), and some actions require embodied resources (energy generation potentially requires water and off‐base feedstock). We suggest that integrating multiple net zero goals can lead to lower environmental impact for military bases.     

Gross primary production controls the subsequent winter CO2 exchange in a boreal peatland

In high‐latitude regions, carbon dioxide (CO₂) emissions during the winter represent an important component of the annual ecosystem carbon budget; however, the mechanisms that control the winter CO₂ emissions are currently not well understood. It has been suggested that substrate availability from soil labile carbon pools is a main driver of winter CO₂ emissions. In ecosystems that are dominated by annual herbaceous plants, much of the biomass produced during the summer is likely to contribute to the soil labile carbon pool through litter fall and root senescence in the autumn. Thus, the summer carbon uptake in the ecosystem may have a significant influence on the subsequent winter CO₂ emissions. To test this hypothesis, we conducted a plot‐scale shading experiment in a boreal peatland to reduce the gross primary production (GPP) during the growing season. At the growing season peak, vascular plant biomass in the shaded plots was half that in the control plots. During the subsequent winter, the mean CO₂ emission rates were 21% lower in the shaded plots than in the control plots. In addition, long‐term (2001–2012) eddy covariance data from the same site showed a strong correlation between the GPP (particularly the late summer and autumn GPP) and the subsequent winter net ecosystem CO₂ exchange (NEE). In contrast, abiotic factors during the winter could not explain the interannual variation in the cumulative winter NEE. Our study demonstrates the presence of a cross‐seasonal link between the growing season biotic processes and winter CO₂ emissions, which has important implications for predicting winter CO₂ emission dynamics in response to future climate change.