Hydrographic Study of Peirce Island Wastewater Treatment Plant Effluent in the Piscataqua River of Portsmouth, New Hampshire: Report of Findings from the December 10 – 14, 2012 Study Period

In order to assist the New Hampshire Department of Environmental Services (NHDES) evaluate the impact of treated wastewater effluent from Peirce Island Wastewater Treatment Plant (WWTP) to the Lower Piscataqua River and Portsmouth Harbor a hydrographic dye study was conducted in December 2012 in Portsmouth, NH. Eight (8) shellfish cages with American oysters (Crassostrea virginica) and blue mussels (Mytilus edulis) were deployed both upstream and downstream of the Peirce Island WWTP in the Piscataqua River, Little Harbor, and the entrance of Little Bay. Eight (8) mini CTDs that monitor conductivity/salinity, temperature, and depth, and six (6) moored fluorometers, which measure dye tagged effluent from the Peirce Island WWTP were attached to the subsurface cages. A fifty (50) gallon mixture of Rhodamine WT dye and distilled water was injected into WWTP on December 11, 2012 for a half tidal cycle (approximately 12.4 hours). Additionally, boat tracking fluorometers connected with a mobile geographic information system (GIS) were used to measure dye levels on the surface in situ and in real time. Microbiological analyses of fecal coliform (FC), male-specific coliphage (MSC), Norovirus (NoV) genogroup I (GI) and genogroup II (GII), and Adenovirus (AdV) were conducted on WWTP influent and effluent composite samples collected with automated samplers to determine the WWTP efficiency in reducing indicator bacteria and viruses. Microbiological sampling and testing of oysters and mussels from the eight (8) sentinel cages was conducted to assess the impact of WWTP effluent on shellfish growing areas and growing area classifications. Prior to conducting the study, the assumption was that the FDA’s recommended minimum dilution of 1000:1was not applicable in this situation because the recommended dilution is based on a WWTP having at least secondary treatment. The microbiological findings in shellfish samples, wastewater samples from the Peirce Island WWTP, and the results of the dye study, confirm that a minimum of 1,000:1 dilution with respect to Peirce Island WWTP is currently not applicable for this WWTP. The FDA and NHDES recommend continued MSC testing of wastewater samples from the WWTP before and after the WWTP upgrade. The FDA and NHDES recommend a future field study after the WWTP upgrade in order to delineate the 1,000:1 dilution zone

Transplantation of Ciliary Neurotrophic Factor-Expressing Adult Oligodendrocyte Precursor Cells Promotes Remyelination and Functional Recovery after SpinalCord Injury

Demyelination contributes to the dysfunction after traumatic spinal cord injury (SCI). We explored whether the combination of neurotrophic factors and transplantation of adult rat spinal cord oligodendrocyte precursor cells (OPCs) could enhance remyelination and functional recovery after SCI. Ciliary neurotrophic factor (CNTF) was the most effective neurotrophic factor to promote oligodendrocyte (OL) differentiation and survival of OPCs in vitro. OPCs were infected with retroviruses expressing enhanced green fluorescent protein (EGFP) or CNTF and transplanted into the contused adult thoracic spinal cord 9 d after injury. Seven weeks after transplantation, the grafted OPCs survived and integrated into the injured spinal cord. The survival of grafted CNTF-OPCs increased fourfold compared with EGFP-OPCs. The grafted OPCs differentiated into adenomatus polyposis coli (APC+) OLs, and CNTF significantly increased the percentage of APC+ OLs from grafted OPCs. Immunofluorescent and immunoelectron microscopic analyses showed that the grafted OPCs formed central myelin sheaths around the axons in the injured spinal cord. The number of OL-remyelinated axons in ventrolateral funiculus (VLF) or lateral funiculus (LF) at the injured epicenter was significantly increased in animals that received CNTF-OPC grafts compared with all other groups. Importantly, 75% of rats receiving CNTF-OPC grafts recovered transcranial magnetic motor-evoked potential and magnetic interenlargement reflex responses, indicating that conduction through the demyelinated axons in VLF or LF, respectively, was partially restored. More importantly, recovery of hindlimb locomotor function was significantly enhanced in animals receiving grafts of CNTF-OPCs. Thus, combined treatment with OPC grafts expressing CNTF can enhance remyelination and facilitate functional recovery after traumatic SCI

An organic-inorganic hybrid scaffold with honeycomb-like structures enabled by one-step self-assembly-driven electrospinning

Electrospun organic/inorganic hybrid scaffolds have been appealing in tissue regeneration owing to the integrated physicochemical and biological performances. However, the conventional electrospun scaffolds with non-woven structures usually failed to enable deep cell infiltration due to the densely stacked layers among the fibers. Herein, through self-assembly-driven electrospinning, a polyhydroxybutyrate/poly(e-caprolactone)/58S sol-gel bioactive glass (PHB/PCL/58S) hybrid scaffold with honeycomb-like structures was prepared by manipulating the solution composition and concentration during a one-step electrospinning process. The mechanisms enabling the formation of self-assembled honeycomb-like structures were investigated through comparative studies using Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) between PHB/PCL/58S and PHB/PCL/sol-gel silica systems. The obtained honeycomb-like structure was built up from nanofibers with an average diameter of 370 nm and showed a bimodal distribution of pores: large polygonal pores up to hundreds of micrometers within the honeycomb-cells and irregular pores among the nanofibers ranging around few micrometers. The cell-materials interactions were further studied by culturing MG-63 osteoblast-like cells for 7 days. Cell viability, cell morphology and cell infiltration were comparatively investigated as well. While cells merely proliferated on the surface of non-woven structures, MG-63 cells showed extensive proliferation and deep infiltration up to 100-200 mu m into the honeycomb-like structure. Moreover, the cellular spatial organization was readily regulated by the honeycomb-like pattern as well. Overall, the newly obtained hybrid scaffold may integrate the enhanced osteogenicity originating from the bioactive components, and the improved cell-material interactions brought by the honeycomb-like structure, making the new scaffold a promising candidate for tissue regeneration.Peer reviewe

Astrocytes from the contused spinal cord inhibit oligodendrocyte differentiation of adult oligodendrocyte precursor cells by increasing the expression of bone morphogenetic proteins.

Promotion of remyelination is an important therapeutic strategy to facilitate functional recovery after traumatic spinal cord injury (SCI). Transplantation of neural stem cells (NSCs) or oligodendrocyte precursor cells (OPCs) has been used to enhance remyelination after SCI. However, the microenvironment in the injured spinal cord is inhibitory for oligodendrocyte (OL) differentiation of NSCs or OPCs. Identifying the signaling pathways that inhibit OL differentiation in the injured spinal cord could lead to new therapeutic strategies to enhance remyelination and functional recovery after SCI. In the present study, we show that reactive astrocytes from the injured rat spinal cord or their conditioned media inhibit OL differentiation of adult OPCs with concurrent promotion of astrocyte differentiation. The expression of bone morphogenetic proteins (BMP) is dramatically increased in the reactive astrocytes and their conditioned media. Importantly, blocking BMP activity by BMP receptor antagonist, noggin, reverse the effects of active astrocytes on OPC differentiation by increasing the differentiation of OL from OPCs while decreasing the generation of astrocytes. These data indicate that the upregulated bone morphogenetic proteins in the reactive astrocytes are major factors to inhibit OL differentiation of OPCs and to promote its astrocyte differentiation. These data suggest that manipulation of BMP signaling in the endogenous or grafted NSCs or OPCs may be a useful therapeutic strategy to increase their OL differentiation and remyelination and enhance functional recovery after SCI

Exploring the ‘middle ground’ between state and market: the example of China

Studies of housing systems lying in the ‘middle ground’ between state and market are subject to three important shortcomings. First, the widely used Esping-Andersen (EA) approach assesses only a subset of the key housing outcomes and may be less helpful for describing changes in housing policy regimes. Second, there is too much emphasis on tenure transitions, and an assumed close correspondence between tenure labels and effective system functioning may not be valid. Third, due attention has not been given to the spatial dimensions in which housing systems operate, in particular when housing policies have a significant devolved or localised emphasis. Updating EA’s framework, we suggest a preliminary list of housing system indicators in order to capture the nature of the housing systems being developed and devolved. We verified the applicability of this indicator system with the case of China. This illustrates clearly the need for a more nuanced and systematic basis for categorising differences and changes in welfare and housing policies

Dynamic changes of cytotoxic T lymphocytes (CTLs), natural killer (NK) cells, and natural killer T (NKT) cells in patients with acute hepatitis B infection

<p>Abstract</p> <p><b>Background</b></p> <p>The goal of this study is to observe changes in HBcAg-specific cytotoxic T lymphocytes (CTLs), natural killer (NK) and natural killer T (NKT) cells from peripheral blood and to relate such changes on viral clearance and liver injury in patients with acute hepatitis B (AHB).</p> <p><b>Methods</b></p> <p>Dynamic profiles on the frequency of HLA-A0201-restricted HBcAg18-27 pentamer complex (MHC-Pentamer)-specific CTLs and lymphocyte subsets in AHB patients were analyzed in addition to liver function tests, HBV serological markers, and HBV DNA levels. ELISPOT was used to detect interferon-gamma (INF-γ) secretion in specific CTLs stimulated with known T cell epitope peptides associated with HBV surface protein, polymerase, and core protein.</p> <p><b>Results</b></p> <p>HBV-specific CTL frequencies in AHB patients were much higher than in patients with chronic hepatitis B (CHB) (p < 0.05). HBeAg and HBV DNA disappeared earlier in AHB patients with a high frequency of HBV-specific CTLs compared with those with a low frequency of HBV-specific CTLs (p = 0.001 and 0.024, respectively). INF-γ spots of effector cells stimulated by Pol575-583, Env348-357, or Core18-27 epitope peptides were significantly greater in AHB patients than in CHB patients (p < 0.01). CD3⁺CD8⁺T cell numbers in AHB patients was more than observed in the healthy control group from the first to the fourth week after admission (p <it>= </it>0.008 and 0.01, respectively); the number of CD3⁺CD8⁺T cells and frequency of HBcAg18-27-specific CTLs in AHB patients reached peak levels at the second week after admission. NK and NKT cell numbers were negatively correlated with the frequency of HBcAg-specific CTLs (<it>r </it>= -0.266, p = 0.05).</p> <p><b>Conclusions</b></p> <p>Patients with AHB possess a higher frequency of HBcAg-specific CTLs than CHB patients. The frequency of specific CTLs in AHB patients is correlated with HBeAg clearance indicating that HBV-specific CTLs play an important role in viral clearance and the self-limited process of the disease. Furthermore, NK and NKT cells are likely involved in the early, non-specific immune response to clear the virus.</p

A type VII-secreted lipase toxin with reverse domain arrangement

Funding: This study was supported by the Wellcome Trust (through Investigator Awards 10183/Z/15/Z and 224151/Z/21/Z to TP), the Intramural Research Program of the NIH, NCI, Center for Cancer Research (awarded to SML), the German Centre of Infection Research (DZIF) to SH (TTU 08.708). NM holds a Walter Benjamin Fellowship (M2871/1-1), funded by the DFG (German Research Foundation). Additionally, we acknowledge infrastructural funding by the DFG in the frame of Germany's Excellence Strategy—EXC 2124—390838134 (SH). SG is funded by the Newcastle-Liverpool-Durham BBSRC DTP2 Training Grant, project reference number BB/M011186/1 and YY by the China Scholarship Council.The type VII protein secretion system (T7SS) is found in many Gram-positive bacteria and in pathogenic mycobacteria. All T7SS substrate proteins described to date share a common helical domain architecture at the N-terminus that typically interacts with other helical partner proteins, forming a composite signal sequence for targeting to the T7SS. The C-terminal domains are functionally diverse and in Gram-positive bacteria such as Staphylococcus aureus often specify toxic anti-bacterial activity. Here we describe the first example of a class of T7 substrate, TslA, that has a reverse domain organisation. TslA is widely found across Bacillota including Staphylococcus, Enterococcus and Listeria. We show that the S. aureus TslA N-terminal domain is a phospholipase A with anti-staphylococcal activity that is neutralised by the immunity lipoprotein TilA. Two small helical partner proteins, TlaA1 and TlaA2 are essential for T7-dependent secretion of TslA and at least one of these interacts with the TslA C-terminal domain to form a helical stack. Cryo-EM analysis of purified TslA complexes indicate that they share structural similarity with canonical T7 substrates. Our findings suggest that the T7SS has the capacity to recognise a secretion signal present at either end of a substrate.Publisher PDFPeer reviewe

Calcium/calmodulin-dependent kinase kinase 2 regulates hematopoietic stem and progenitor cell regeneration

Hematopoietic stem and progenitor cells (HSPCs) are predominantly quiescent in adults, but proliferate in response to bone marrow (BM) injury. Here, we show that deletion of Ca2+/calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2) promotes HSPC regeneration and hematopoietic recovery following radiation injury. Using Camkk2-enhanced green fluorescent protein (EGFP) reporter mice, we found that Camkk2 expression is developmentally regulated in HSPC. Deletion of Camkk2 in HSPC results in a significant downregulation of genes affiliated with the quiescent signature. Accordingly, HSPC from Camkk2 null mice have a high proliferative capability when stimulated in vitro in the presence of BM-derived endothelial cells. In addition, Camkk2 null mice are more resistant to radiation injury and show accelerated hematopoietic recovery, enhanced HSPC regeneration and ultimately a prolonged survival following sublethal or lethal total body irradiation. Mechanistically, we propose that CaMKK2 regulates the HSPC response to hematopoietic damage by coupling radiation signaling to activation of the anti-proliferative AMP-activated protein kinase. Finally, we demonstrated that systemic administration of the small molecule CaMKK2 inhibitor, STO-609, to irradiated mice enhanced HSPC recovery and improved survival. These findings identify CaMKK2 as an important regulator of HSPC regeneration and demonstrate CaMKK2 inhibition is a novel approach to promoting hematopoietic recovery after BM injury

Joint profiling of DNA methylation and chromatin architecture in single cells.

We report a molecular assay, Methyl-HiC, that can simultaneously capture the chromosome conformation and DNA methylome in a cell. Methyl-HiC reveals coordinated DNA methylation status between distal genomic segments that are in spatial proximity in the nucleus, and delineates heterogeneity of both the chromatin architecture and DNA methylome in a mixed population. It enables simultaneous characterization of cell-type-specific chromatin organization and epigenome in complex tissues