Role of phospholipase A2 (group I secreted) in the genesis of basal tone in the internal anal sphincter smooth muscle

The role of phospholipase A(2) (PLA(2)) in the genesis of basal tone in the internal anal sphincter (IAS) is not known. We determined the effects of PLA(2) and inhibitors on the basal tone and intraluminal pressures (IASP) in the rat IAS vs. rectal smooth muscles (RSM). In addition, we determined the correlations between the IAS tone, PLA(2) levels, and the actual enzymatic activity. Inhibition of PLA(2) by 4-bromophenacyl bromide (universal inhibitor of PLA(2)) and MJ33 [selective inhibitor of secreted isoform of PLA(2) (sPLA(2))] caused concentration-dependent decrease in the IAS tone and in the IASP. Maximal decreases in the IAS tone and IASP by 4-bromophenacyl bromide and MJ33 were 58.8 +/- 6.9 and 51.5 +/- 6.3%, and 66.7 +/- 5.1 and 79.8 +/- 8.2%, respectively. The sPLA(2) inhibitors were approximately 100 times more potent in decreasing the IASP than the mean blood pressure. Conversely, the selective inhibitors of the cytosolic and calcium-independent PLA(2) arachidonyl trifluoromethyl ketone and bromoenol lactone, respectively, produced no significant effect. The IAS had characteristically higher levels of sPLA(2) activity (26.5 +/- 4.9 micromol.min(-1).ml(-1)) vs. the RSM (3.2 +/- 0.4 micromol.min(-1).ml(-1)), and higher levels of sPLA(2) as shown by Western blot and RT-PCR. Interestingly, administration of sPLA(2) transformed RSM into the tonic smooth muscle like that of the IAS: it developed basal tone and relaxed in response to the electrical field stimulation. From the present data, we conclude that sPLA(2) plays a critical role in the genesis of tone in the IAS. PLA(2) inhibitors may provide potential therapeutic target for treating anorectal motility disorders.     

Integration of gene-based markers in a pearl millet genetic map for identification of candidate genes underlying drought tolerance quantitative trait loci

Background  

Identification of genes underlying drought tolerance (DT) quantitative trait loci (QTLs) will facilitate understanding of molecular mechanisms of drought tolerance, and also will accelerate genetic improvement of pearl millet through marker-assisted selection. We report a map based on genes with assigned functional roles in plant adaptation to drought and other abiotic stresses and demonstrate its use in identifying candidate genes underlying a major DT-QTL.     

Fungal Infections in COVID-19 Intensive Care Patients

Opportunistic fungal infections increase morbidity and mortality in COVID-19 patients monitored in intensive care units (ICU). As patients’ hospitalization days in the ICU and intubation period increase, opportunistic infections also increase, which prolongs hospital stay days and elevates costs. The study aimed to describe the profile of fungal infections and identify the risk factors associated with mortality in COVID-19 intensive care patients. The records of 627 patients hospitalized in ICU with the diagnosis of COVID-19 were investigated from electronic health records and hospitalization files. The demographic characteristics (age, gender), the number of ICU hospitalization days and mortality rates, APACHE II scores, accompanying diseases, antibiotic-steroid treatments taken during hospitalization, and microbiological results (blood, urine, tracheal aspirate samples) of the patients were recorded. Opportunistic fungal infection was detected in 32 patients (5.10%) of 627 patients monitored in ICU with a COVID-19 diagnosis. The average APACHE II score of the patients was 28 ± 6. While 25 of the patients (78.12%) died, seven (21.87%) were discharged from the ICU. Candida parapsilosis (43.7%) was the opportunistic fungal agent isolated from most blood samples taken from COVID-19 positive patients. The mortality rate of COVID-19 positive patients with candidemia was 80%. While two out of the three patients (66.6%) for whom fungi were grown from their tracheal aspirate died, one patient (33.3%) was transferred to the ward. Opportunistic fungal infections increase the mortality rate of COVID-19-positive patients. In addition to the risk factors that we cannot change, invasive procedures should be avoided, constant blood sugar regulation should be applied, and unnecessary antibiotics use should be avoided.     

Carbon Sequestration in Reclaimed Minesoils

Minesoils are drastically influenced by anthropogenic activities. They are characterized by low soil organic matter (SOM) content, low fertility, and poor physicochemical and biological properties, limiting their quality, capability, and functions. Reclamation of these soils has potential for resequestering some of the C lost and mitigating CO₂ emissions. Soil organic carbon (SOC) sequestration rates in minesoils are high in the first 20 to 30 years after reclamation in the top 15 cm soil depth. In general, higher rates of SOC sequestration are observed for minesoils under pasture and grassland management than under forest land use. Observed rates of SOC sequestration are 0.3 to 1.85 Mg C ha^{? 1} yr^{? 1} for pastures and rangelands, and 0.2 to 1.64 Mg C ha^{? 1} yr^{? 1} for forest land use. Proper reclamation and postreclamation management may enhance SOC sequestration and add to the economic value of the mined sites. Management practices that may enhance SOC sequestration include increasing vegetative cover by deep-rooted perennial vegetation and afforestation, improving soil fertility, and alleviation of physical, chemical and biological limitations by fertilizers and soil amendments such as biosolids, manure, coal combustion by-products, and mulches. Soil and water conservation are important to SOC sequestration. The potential of SOC sequestration in minesoils of the US is estimated to be 1.28 Tg C yr^?1, compared to the emissions from coal combustion of 506 Tg C yr^{? 1}.     

Increased molecular damage and heterogeneity as the basis of aging

Aging at the molecular level is characterized by the progressive accumulation of molecular damage. The sources of damage act randomly through environmental and metabolically generated free radicals, through spontaneous errors in biochemical reactions, and through nutritional components. However, damage to a macromolecule may depend on its structure, localization and interactions with other macromolecules. Damage to the maintenance and repair pathways comprising homeodynamic machinery leads to age-related failure of homeodynamics, increased molecular heterogeneity, altered cellular functioning, reduced stress tolerance, diseases and ultimate death. Novel approaches for testing and developing effective means of intervention, prevention and modulation of aging involve means to minimize the occurrence and accumulation of molecular damage. Mild stress-induced hormesis by physical, biological and nutritional methods, including hormetins, represents a promising strategy for achieving healthy aging and for preventing age-related diseases.     

Bioethanol Potentials and Life-Cycle Assessments of Biofuel Feedstocks

A wide range of bioenergy crops has been proposed as feedstocks that can serve as renewable and ecologically sound substitutes to fossil fuels. In the United States, corn grain (Zea mays) ethanol is the primary biofuel, with over 49 billion liters produced in 2010. Along with the Energy Independence and Security Act (EISA) of 2007 mandate, concerns about competition for food, land availability, nutrient and water requirements, energy balances, and greenhouse gas (GHG) emissions have prompted researchers to investigate other potential feedstocks. These include second-generation lignocellulosic feedstock and third-generation biodiesel from microalgae and cyanobacteria. However, each feedstock option has associated benefits and consequences for its use. One technique used to evaluate the energy efficiency of bioenergy production systems is the life-cycle assessment (LCA), where system inputs and outputs are computed in terms of either C or energy equivalents to assess the net gains in energy or C offsets. This article collates and synthesizes information about feedstock production options. Results show a wide range of calculated energy and GHG balances, even for the same feedstock species. Discrepancies in LCA and uncertainty thus make direct comparisons difficult and prevent a consensus in determining feedstock suitability. Recommendations must be based upon LCA model assumptions, crop species, cultivation methods, management practices, and energy conversion choices. Currently lignocellulosic feedstock, while a better alternative than corn grain, is not a long-term viable energy source. New feedstocks and technologies are necessary if bioenergy is to be C-neutral and efficient in energy production and land use. Although C fluxes are considered in LCA, one important ecosystem C stock that has previously been left out of many LCA models is changes to soil organic carbon (SOC). Future research, developments, and priorities are discussed for options to produce low C fuel sources and stabilize the climate.     

Changes in succinate dehydrogenase activity of rats after intraventricular injections of GABA, muscimol and picrotoxin

Effects of intraventricular injections of GABA, and a GABA agonist, muscimol and an antagonist, picrotoxin on succinate dehydrogenase (SDH) enzyme activity in plasma and a few hypothalamic nuclei of brain of rats have been investigated using biochemical, histochemical and cytophotometric techniques. Results show that SDH decreased by GABA and muscimol treatment, and increased after picrotoxin injection. From the above findings, it is apparent that GABA, muscimol and picrotoxin influence SDH activity of plasma and hypothalamic nuclei.     

Comparison of Soil Quality Index Using Three Methods

Assessment of management-induced changes in soil quality is important to sustaining high crop yield. A large diversity of cultivated soils necessitate identification development of an appropriate soil quality index (SQI) based on relative soil properties and crop yield. Whereas numerous attempts have been made to estimate SQI for major soils across the World, there is no standard method established and thus, a strong need exists for developing a user-friendly and credible SQI through comparison of various available methods. Therefore, the objective of this article is to compare three widely used methods to estimate SQI using the data collected from 72 soil samples from three on-farm study sites in Ohio. Additionally, challenge lies in establishing a correlation between crop yield versus SQI calculated either depth wise or in combination of soil layers as standard methodology is not yet available and was not given much attention to date. Predominant soils of the study included one organic (Mc), and two mineral (CrB, Ko) soils. Three methods used to estimate SQI were: (i) simple additive SQI (SQI-1), (ii) weighted additive SQI (SQI-2), and (iii) statistically modeled SQI (SQI-3) based on principal component analysis (PCA). The SQI varied between treatments and soil types and ranged between 0–0.9 (1 being the maximum SQI). In general, SQIs did not significantly differ at depths under any method suggesting that soil quality did not significantly differ for different depths at the studied sites. Additionally, data indicate that SQI-3 was most strongly correlated with crop yield, the correlation coefficient ranged between 0.74–0.78. All three SQIs were significantly correlated (r = 0.92–0.97) to each other and with crop yield (r = 0.65–0.79). Separate analyses by crop variety revealed that correlation was low indicating that some key aspects of soil quality related to crop response are important requirements for estimating SQI.     

The phylogenetic potential of entire 26S rDNA sequences in plants

18S ribosomal RNA genes are the most widely used nuclear sequences for phylogeny reconstruction at higher taxonomic levels in plants. However, due to a conservative rate of evolution, 18S rDNA alone sometimes provides too few phylogenetically informative characters to resolve relationships adequately. Previous studies using partial sequences have suggested the potential of 26S or large-subunit (LSU) rDNA for phylogeny retrieval at taxonomic levels comparable to those investigated with 18S rDNA. Here we explore the patterns of molecular evolution of entire 26S rDNA sequences and their impact on phylogeny retrieval. We present a protocol for PCR amplification and sequencing of entire (approximately 3.4 kb) 26S rDNA sequences as single amplicons, as well as primers that can be used for amplification and sequencing. These primers proved useful in angiosperms and Gnetales and likely have broader applicability. With these protocols and primers, entire 26S rDNA sequences were generated for a diverse array of 15 seed plants, including basal eudicots, monocots, and higher eudicots, plus two representatives of Gnetales. Comparisons of sequence dissimilarity indicate that expansion segments (or divergence domains) evolve 6.4 to 10.2 times as fast as conserved core regions of 26S rDNA sequences in plants. Additional comparisons indicate that 26S rDNA evolves 1.6 to 2.2 times as fast as and provides 3.3 times as many phylogenetically informative characters as 18S rDNA; compared to the chloroplast gene rbcL, 26S rDNA evolves at 0.44 to 1.0 times its rate and provides 2.0 times as many phylogenetically informative characters. Expansion segment sequences analyzed here evolve 1.2 to 3.0 times faster than rbcL, providing 1.5 times the number of informative characters. Plant expansion segments have a pattern of evolution distinct from that found in animals, exhibiting less cryptic sequence simplicity, a lower frequency of insertion and deletion, and greater phylogenetic potential.      

Reduced levels of ADP-ribosylatable elongation factor-2 in aged and SV40-transformed human cell cultures

The elongation step is involved in the regulation of protein synthesis during the cell cycle, environmental stress, ageing and transformation. Using a diphtheria toxin-mediated assay for measuring the levels of ADP-ribosylatable elongation factor EF-2, we have observed an irreversible decrease of up to 64% in the amount of ADP-ribosylatable EF-2 in normal diploid human fibroblasts MRC-5 undergoing ageing in vitro. However, a similar decrease in low serum-associated G0/G1-arrested cells is reversible both in MRC-5 cells and in their SV40-transformed counterparts. Reduced levels of ADP-ribosylatable EF-2 could account for the slowing-down of protein synthesis during cell cycle arrest and during cellular ageing in culture.