Matrix metalloproteinases are involved in C-terminal and interglobular domain processing of cartilage aggrecan in late stage cartilage degradation.

Monoclonal antibody (MAb) technology was used to examine aggrecan metabolites and the role of aggrecanases and matrix metalloproteinases (MMPs) in proteolysis of the interglobular domain (IGD) and C-terminus of aggrecan. An in vitro model of progressive cartilage degradation characterized by early proteoglycan loss and late stage collagen catabolism was evaluated in conjunction with a broad-spectrum inhibitor of MMPs. We have for the first time demonstrated that IGD cleavage by MMPs occurs during this late stage cartilage degeneration, both as a primary event in association with glycosaminoglycan (GAG) release from the tissue and secondarily in trimming of aggrecanase-generated G1 metabolites. Additionally, we have shown that MMPs were responsible for C-terminal catabolism of aggrecan and generation of chondroitin sulfate (CS) deficient aggrecan monomers and that this aggrecan truncation occurred prior to detectable IGD cleavage by MMPs. The onset of this later stage MMP activity was also evident by the generation of MMP-specific link protein catabolites in this model culture system. Recombinant MMP-1, -3 and -13 were all capable of C-terminally truncating aggrecan with at least two cleavage sites N-terminal to the CS attachment domains of aggrecan. Through analysis of aggrecan metabolites in pathological synovial fluids from human, canine and equine sources, we have demonstrated the presence of aggrecan catabolites that appear to have resulted from similar C-terminal processing of aggrecan as that induced in our in vitro culture systems. Finally, by developing a new MAb recognizing a linear epitope in the IGD of aggrecan, we have identified two novel aggrecan metabolites generated by an as yet unidentified proteolytic event. Collectively, these results suggest that C-terminal processing of aggrecan by MMPs may contribute to the depletion of cartilage GAG that leads to loss of tissue function in aging and disease. Furthermore, analysis of aggrecan metabolites resulting from both C-terminal and IGD cleavage by MMPs may prove useful in monitoring different stages in the progression of cartilage degeneration.     

Sexual dimorphism in healthy aging and mild cognitive impairment: a DTI study

Previous PET and MRI studies have indicated that the degree to which pathology translates into clinical symptoms is strongly dependent on sex with women more likely to express pathology as a diagnosis of AD, whereas men are more resistant to clinical symptoms in the face of the same degree of pathology. Here we use DTI to investigate the difference between male and female white matter tracts in healthy older participants (24 women, 16 men) and participants with mild cognitive impairment (21 women, 12 men). Differences between control and MCI participants were found in fractional anisotropy (FA), radial diffusion (DR), axial diffusion (DA) and mean diffusion (MD). A significant main effect of sex was also reported for FA, MD and DR indices, with male control and male MCI participants having significantly more microstructural damage than their female counterparts. There was no sex by diagnosis interaction. Male MCIs also had significantly less normalised grey matter (GM) volume than female MCIs. However, in terms of absolute brain volume, male controls had significantly more brain volume than female controls. Normalised GM and WM volumes were found to decrease significantly with age with no age by sex interaction. Overall, these data suggest that the same degree of cognitive impairment is associated with greater structural damage in men compared with women.     

Sensitivity of yeast strains with long G-tails to levels of telomere-bound telomerase

The Saccharomyces cerevisiae Pif1p helicase is a negative regulator of telomere length that acts by removing telomerase from chromosome ends. The catalytic subunit of yeast telomerase, Est2p, is telomere associated throughout most of the cell cycle, with peaks of association in both G1 phase (when telomerase is not active) and late S/G2 phase (when telomerase is active). The G1 association of Est2p requires a specific interaction between Ku and telomerase RNA. In mutants lacking this interaction, telomeres were longer in the absence of Pif1p than in the presence of wild-type PIF1, indicating that endogenous Pif1p inhibits the active S/G2 form of telomerase. Pif1p abundance was cell cycle regulated, low in G1 and early S phase and peaking late in the cell cycle. Low Pif1p abundance in G1 phase was anaphase-promoting complex dependent. Thus, endogenous Pif1p is unlikely to act on G1 bound Est2p. Overexpression of Pif1p from a non-cell cycle-regulated promoter dramatically reduced viability in five strains with impaired end protection (cdc13–1, yku80Δ, yku70Δ, yku80–1, and yku80–4), all of which have longer single-strand G-tails than wild-type cells. This reduced viability was suppressed by deleting the EXO1 gene, which encodes a nuclease that acts at compromised telomeres, suggesting that the removal of telomerase by Pif1p exposed telomeres to further C-strand degradation. Consistent with this interpretation, depletion of Pif1p, which increases the amount of telomere-bound telomerase, suppressed the temperature sensitivity of yku70Δ and cdc13–1 cells. Furthermore, eliminating the pathway that recruits Est2p to telomeres in G1 phase in a cdc13–1 strain also reduced viability. These data suggest that wild-type levels of telomere-bound telomerase are critical for the viability of strains whose telomeres are already susceptible to degradation.     

Pollution characteristics and health risk assessment of potentially toxic elements in school playground soils: A case study of Lagos,Nigeria

Abstract

Soil samples were collected in February 2014 from 25 school playgrounds in Lagos, Nigeria to assess the potential adverse effects of the exposure of children to potentially toxic element (PTE). In each of the playgrounds, about 500?g of bulked soil samples were collected, dried, sieved, acid digested, and analyzed by inductively coupled plasma mass spectrometry (ICP‐MS). Results showed that soils studied were characteristically unpolluted as the average PTE concentration at each site did not exceed the soil guideline values. Considering the pollution assessment tools employed, some soil samples showed some form of anthropogenic input from PTE. Health risk assessment was employed to assess PTE exposure from ingestion, inhalation, and dermal contact. Result indicated that the highest risk is associated with ingestion followed by dermal contact and inhalation. For non‐carcinogenic effects, exposure to school playground soils poses no threat to children as the overall value of hazard index is less than the safe level of 1. For carcinogenic effects, only Cr and Ni were considered and were below the threshold of 1?×?10^–6. This study has demonstrated that minimal risk arises from the investigated playgrounds and that regular monitoring is required to keep the PTE contents low in soils to avoid risk to human health.

Abbreviations: SH, school playground; PTE, potentially toxic elements     

Production of Hyperalgesic Prostaglandins by Sympathetic Postganglionic Neurons

Prostaglandin E2 and prostacyclin (prostaglandin I2) produce hyperalgesia in animals and humans. Because there is evidence that prostaglandins contribute to pain maintained by sympathetic nervous system activity, we evaluated whether sympathetic postganglionic neurons synthesize these hyperalgesic prostaglandins, and whether production of prostaglandins by these neurons can contribute to sensitization of primary afferent nociceptors. Intradermal injection of arachidonic acid but not linoleic acid, in the rat hindpaw, produces a decrease in mechanical nociceptive threshold. This hyperalgesic effect is prevented by indomethacin, an inhibitor of prostaglandin synthesis or by prior surgical removal of the lumbar sympathetic chain. To test the hypothesis that sympathetic postganglionic neurons are the source of prostaglandins, we measured production of prostaglandin E2 and 6-keto-prostaglandin F1 alpha (the stable metabolite of prostacyclin) by homogenates of adult rat sympathetic postganglionic neurons from superior cervical ganglia. These homogenates produced significant amounts of prostaglandin E2 and 6-keto-prostaglandin F1 alpha, and most of this production is eliminated by neonatal administration of 6-hydroxydopamine which selectively destroys sympathetic postganglionic neurons. These results demonstrate that sympathetic postganglionic neurons produce prostaglandins, and supports further the hypothesis that the release of prostaglandins from sympathetic postganglionic neurons contributes to the hyperalgesia associated with sympathetically maintained pain.     

Development of a pigtail macaque model of sexually transmitted infection/HIV coinfection using Chlamydia trachomatis, Trichomonas vaginalis, and SHIV(SF162P3)

Background Sexually transmitted infections (STIs) are associated with an increased risk of HIV infection. To model the interaction between STIs and HIV infection, we evaluated the capacity of the pigtail macaque model to sustain triple infection with Trichomonas vaginalis, Chlamydia trachomatis, and SHIV_SF162P3. Methods Seven SHIV_SF162P3‐infected pigtail macaques were inoculated with T. vaginalis only (n = 2), C. trachomatis only (n = 1), both T. vaginalis and C. trachomatis (n = 2), or control media (no STI; n = 2). Infections were confirmed by culture and/or nucleic acid testing. Genital mucosa was visualized by colposcopy. Results Characteristic gynecologic signs were observed for both STIs, but not in control animals. Manifestations were most prominent at days 7–10 post‐infection. STIs persisted between 4 and 6 weeks and were cleared with antibiotics. Conclusions These pilot studies demonstrate the first successful STI‐SHIV triple infection of pigtail macaques, with clinical presentation of genital STI symptoms similar to those observed in humans.     

A review of thermochemical upgrading of pyrolysis bio‐oil: Techno‐economic analysis,life cycle assessment,and technology readiness

Technologies for upgrading fast pyrolysis bio‐oil to drop‐in fuels and coproducts are under development and show promise for decarbonizing energy supply for transportation and chemicals markets. The successful commercialization of these fuels and the technologies deployed to produce them depend on production costs, scalability, and yield. To meet environmental regulations, pyrolysis‐based biofuels need to adhere to life cycle greenhouse gas intensity standards relative to their petroleum‐based counterparts. We review literature on fast pyrolysis bio‐oil upgrading and explore key metrics that influence their commercial viability through life cycle assessment (LCA) and techno‐economic analysis (TEA) methods together with technology readiness level (TRL) evaluation. We investigate the trade‐offs among economic, environmental, and technological metrics derived from these methods for individual technologies as a means of understanding their nearness to commercialization. Although the technologies reviewed have not attained commercial investment, some have been pilot tested. Predicting the projected performance at scale‐up through models can, with industrial experience, guide decision‐making to competitively meet energy policy goals. LCA and TEA methods that ensure consistent and reproducible models at a given TRL are needed to compare alternative technologies. This study highlights the importance of integrated analysis of multiple economic, environmental, and technological metrics for understanding performance prospects and barriers among early stage technologies.     

A comparative study of the wild and mutated heavy metal resistant Klebsiella variicola generated for cadmium bioremediation

Ten indigenous heavy metals resistant bacteria were isolated from the discharged effluent of Biological Sciences building at Obafemi Awolowo University, Ile-Ife, Osun State, Nigeria. The bacterial strains were isolated using enrichment method in tryptone soya agar (TSA) supplemented with 100 mg/L of Pb, Cd, As, and Ni. All the isolated bacteria showed multiple tolerances to the heavy metals; however, strain GBB 220, which showed a maximum tolerable concentrations (MTCs) of 2000, 1200, 4700, and 1000 mg/L for Pb, Cd, As, and Ni, respectively, was selected for further studies. The bacterium was identified by 16S rRNA sequences as Klebsiella variicola and was subjected to mutational enhancement by acridine orange and ethidium bromide. Eight mutants were recovered, strains K. variicola MutAa–Ad showed improvement in their MTCs of 2500, 2200, and 5000 mg/L for Pb, Cd, and As whereas K. variicola MutEa–Ed had same MTCs as the wild type except for cadmium which increased from 1200 to 1500 mg/L. The antibiotic susceptibility patterns showed that all the strains of K. variicola had multiple resistances to some of the antibiotics. The K. variicola mutants had their optimum pH at 5 and 8 while the wild type had optimum pH at 7 and 8. The cadmium removal efficiency of 97.9% at pH 7, 97.7% at pH 8, and 99.4% at pH 7 was observed for K. variicola Wt, K. variicola Mut Eb, and K. variicola Mut Ac, respectively. These findings suggested that the artificially mutated strains of K. variicola may be applied to remove cadmium from polluted environment.     

Multiple indices of diffusion identifies white matter damage in mild cognitive impairment and Alzheimer's disease

The study of multiple indices of diffusion, including axial (DA), radial (DR) and mean diffusion (MD), as well as fractional anisotropy (FA), enables WM damage in Alzheimer's disease (AD) to be assessed in detail. Here, tract-based spatial statistics (TBSS) were performed on scans of 40 healthy elders, 19 non-amnestic MCI (MCIna) subjects, 14 amnestic MCI (MCIa) subjects and 9 AD patients. Significantly higher DA was found in MCIna subjects compared to healthy elders in the right posterior cingulum/precuneus. Significantly higher DA was also found in MCIa subjects compared to healthy elders in the left prefrontal cortex, particularly in the forceps minor and uncinate fasciculus. In the MCIa versus MCIna comparison, significantly higher DA was found in large areas of the left prefrontal cortex. For AD patients, the overlap of FA and DR changes and the overlap of FA and MD changes were seen in temporal, parietal and frontal lobes, as well as the corpus callosum and fornix. Analysis of differences between the AD versus MCIna, and AD versus MCIa contrasts, highlighted regions that are increasingly compromised in more severe disease stages. Microstructural damage independent of gross tissue loss was widespread in later disease stages. Our findings suggest a scheme where WM damage begins in the core memory network of the temporal lobe, cingulum and prefrontal regions, and spreads beyond these regions in later stages. DA and MD indices were most sensitive at detecting early changes in MCIa.