Role of glycolytically generated ATP for CaMKII-mediated regulation of intracellular Ca2+ signaling in bovine vascular endothelial cells

The role of glycolytically generated ATP in Ca²⁺/calmodulin-dependent kinase II (CaMKII)-mediated regulation of intracellular Ca²⁺ signaling was examined in cultured calf pulmonary artery endothelial (CPAE) cells. Exposure of cells (extracellular Ca²⁺ concentration = 2 mM) to glycolytic inhibitors 2-deoxy-D-glucose (2-DG), pyruvate (pyr) + -hydroxybutyrate (-HB), or iodoacetic acid (IAA) caused an increase of intracellular Ca²⁺ concentration ([Ca²⁺]_i). CaMKII inhibitors (KN-93, W-7) triggered a similar increase of [Ca²⁺]_i. The rise of [Ca²⁺]_i was characterized by a transient spike followed by a small sustained plateau of elevated [Ca²⁺]_i. In the absence of extracellular Ca²⁺ 2-DG caused an increase in [Ca²⁺]_i, suggesting that inhibition of glycolysis directly triggered release of Ca²⁺ from intracellular endoplasmic reticulum (ER) Ca²⁺ stores. The inositol-1,4,5-trisphosphate receptor (IP₃R) inhibitor 2-aminoethoxydiphenyl borate abolished the KN-93- and 2-DG-induced Ca²⁺ response. Ca²⁺ release was initiated in peripheral cytoplasmic processes from which activation propagated as a [Ca²⁺]_i wave toward the central region of the cell. Focal application of 2-DG resulted in spatially confined elevations of [Ca²⁺]_i. Propagating [Ca²⁺]_i waves were preceded by [Ca²⁺]_i oscillations and small, highly localized elevations of [Ca²⁺]_i (Ca²⁺ puffs). Inhibition of glycolysis with 2-DG reduced the KN-93-induced Ca²⁺ response, and vice versa during inhibition of CaMKII 2-DG-induced Ca²⁺ release was attenuated. Similar results were obtained with pyr + -HB and W-7. Furthermore, 2-DG and IAA caused a rapid increase of intracellular Mg²⁺ concentration, indicating a concomitant drop of cellular ATP levels. In conclusion, CaMKII exerts a profound inhibition of ER Ca²⁺ release in CPAE cells, which is mediated by glycolytically generated ATP, possibly through ATP-dependent phosphorylation of the IP₃R. Ca²⁺/calmodulin-dependent kinase II; glycolysis; calcium regulation     

Switching between microtubule- and actin-based transport systems in melanophores is controlled by cAMP levels

BACKGROUND: Intracellular transport involves the movement of organelles along microtubules (MTs) or actin filaments (AFs) by means of opposite-polarity MT motors or actin-dependent motors of the myosin family. The correct delivery of organelles to their different destinations involves a precise coordination of the two transport systems. Such coordination could occur through regulation of the densities of the two cytoskeletal systems or through regulation of the activities of the cytoskeletal motors by signaling mechanisms. RESULTS: To investigate the mechanisms of switching between MT and AF-dependent transport, we examine the influence of the densities of the MT and AF network on pigment transport in fish melanophores. We also change signaling by using activators and inhibitors of Protein Kinase A (PKA). We find that the key parameters characterizing pigment granule transport along MTs do not depend on MT density and are not significantly altered by complete disruption of AFs. In contrast, the kinetics of changes in these parameters correlate with the kinetics of changes in the intracellular levels of cAMP and are affected by the inhibitors of PKA, suggesting the regulation of MT- and AF-dependent motors by cAMP-induced signaling. Furthermore, perturbation of cAMP levels prevents the transfer of pigment granules from MTs onto AFs. CONCLUSIONS: We conclude that the switching of pigment granules between the two major cytoskeletal systems is independent of the densities of MT or AF but is tightly controlled by signaling events.     

Silver-gold alloy nanoparticles biofabricated by fungal xylanases exhibited potent biomedical and catalytic activities

The search for biocompatible nanoparticles with vast applicability has impacted on exploration of various biomaterials for the synthesis of mono and bimetallic nanoparticles. Xylanase is widely regarded as an industrially important enzyme but its potentials in nanotechnological applications are yet to be fully explored. The current study investigates the exploit of xylanases of Aspergillus niger L3 (NE) and Trichoderma longibrachiatum L2 (TE) produced through valorization of corn-cob, to synthesize silver-gold alloy nanoparticles (Ag-AuNPs). Characterization of the Ag-AuNPs involved UV–vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), and field emission scanning electron microscopy and transmission electron microscopy, while their prospective use as antimicrobial, antioxidant, catalytic, anticoagulant, and thrombolytic agents were studied. The biosynthesized Ag-AuNPs were ruby red and light purple with surface plasmon resonance at 520 and 534 nm for NEAg-AuNPs and TEAg-AuNPs, respectively; while FTIR showed that protein molecules capped and stabilized the nanoparticles. The Ag-AuNPs were anisotropic with spherical, oval, and irregular shapes having sizes ranging from 6.98 to 52.51 nm. The nanoparticles appreciably inhibited the growth of tested clinical bacteria (23.40–90.70%) and fungi (70.10–89.05%), and also scavenged 2,2-diphenyl-1-picrylhydrazyl (48.51–53.79%) and hydrogen peroxide (80.5–95.50%). Furthermore, the Ag-AuNPs degraded malachite green (91.39%) and methylene blue (47.10%). Moreover, the Ag-AuNPs displayed outstanding anticoagulant and thrombolytic activities using human blood. This study further emphasizes the significance of xylanases in nanobiotechnology as it has established the potential of xylanases to synthesize Ag-AuNPs, which is being reported for the first time.     

Cervical cytology of recurrent psammoma bodies with atypical cells in a postmenopausal woman: a case report

BACKGROUND: Psammoma bodies (PBs) are an unusual finding in cervical cytology preparations. They have been identified in association with a wide range of benign and malignant conditions within the female genital tract. Portents of a significant underlying pathology include their occurrence in postmenopausal patients, the presence of unexplained vaginal bleeding and their occurrence in association with atypical cells. CASE: PBs associated with atypical cells were detected in repeated cervical cytology smears of an asymptomatic, 55-year-old postmenopausal woman over a 4-year period. She was extensively investigated, and, in the absence of a bleeding) have definitive cause, she underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy. Histologic examination of the specimen demonstrated the presence of bilateral benign ovarian serous cystadenofibromas with large numbers of PBs. Focally the PBs were closely apposed to the serosal surface and invested in a blanket of mesothelial cells. CONCLUSION: Caution is required when assessing the significance of PBs associated with atypical cells in a cervical cytology specimen. Our case demonstrates the presence of ovarian mesothelial cells mimicking atypical glandular cells.     

Species Distribution Modelling predicts habitat suitability and reduction of suitable habitat under future climatic scenario for Sclerophrys perreti: A critically endangered Nigerian endemic toad

Sclerophrys perreti is a critically endangered Nigerian native frog currently imperilled by human activities. A better understanding of its potential distribution and habitat suitability will aid in conservation; however, such knowledge is limited for S. perreti. Herein, we used a species distribution model (SDM) approach with all known occurrence data (n = 22) from our field surveys and primary literature, and environmental variable predictors (19 bioclimatic variables, elevation and land cover) to elucidate habitat suitability and impact of climate change on this species. The SDM showed that temperature and precipitation were the predictors of habitat suitability for S. perreti with precipitation seasonality as the strongest predictor of habitat suitability. The following variable also had a significant effect on habitat suitability: temperature seasonality, temperature annual range, precipitation of driest month, mean temperature of wettest quarter and isothermality. The model predicted current suitable habitat for S. perreti covering an area of 1,115 km². However, this habitat is predicted to experience 60% reduction by 2050 owing to changes in temperature and precipitation. SDM also showed that suitable habitat exists in south-eastern range of the inselberg with predicted low impact of climate change compared to other ranges. Therefore, this study recommends improved conservation measures through collaborations and stakeholder's meeting with local farmers for the management and protection of S. perreti.     

Amphibian assemblages and diversity patterns in two forest ecosystems of South-Eastern Nigeria

Despite recent anthropogenic interference, south-eastern (SE) Nigeria historically hosts a myriad of pristine forest habitats that could sustain diverse amphibian assemblages. Currently, land conversion of primary forests through human activities has led to modified habitats, such as agricultural fields and secondary forest. This calls for biodiversity documentation and initiation of conservation plans. However, the dearth of herpetological studies has limited knowledge of the amphibian species richness and distribution patterns in some forest ecosystems of SE Nigeria. Thus, we surveyed two less-explored intact forests in SE Nigeria—Afi Forest Reserve (AFR) and Ikpan forest ecosystem (IFE)—to document the patterns of amphibian diversity and distribution. Specimens were searched for opportunistically using visual and acoustic survey methods. Literature searches were conducted to document additional species that were not encountered. Using the observational records and arrays of diversity indices, the patterns of species richness and abundance across habitat types were evaluated. The result of our study revealed the presence of 30 and 31 amphibian species from AFR and IFE, respectively. Our field collections included two unidentified species (Sclerophrys sp. and Hyperolius sp.) from AFR and one (Phrynobatrachus sp.) from IFE. Analyses revealed high species diversity and richness in forest and swamps compared with the agricultural field and savannah. Given the increased human activities and the antecedent threats to forest ecosystems of SE Nigeria, we offer recommendations for the initiation of conservation plans for these two surveyed localities.     

Survey of aflatoxin M1 in cows’ milk from free-grazing cows in Abeokuta,Nigeria

Aflatoxin M₁ (AFM₁) in milk from 100 different herds of free-grazing cows in Abeokuta, Nigeria, was analysed by immunoaffinity column cleanup and HPLC with fluorescence detection. AFM₁ was found in 75 % of the samples, the toxin levels in positive samples ranged from 9.0 to 456.0 ng/l. The mean AFM₁ level in positive samples was 108.15 ng/l, exceeding, for example, the European Union maximum level by a factor of two. These results indicated that there is an urgent need to more closely control the milk of free-grazing cows for AFM₁ in order to protect the health of humans consuming milk and milk products.