Microbial removal of nutrients from anaerobic digestate: assessing product-coupled and non-product-coupled approaches.

Although anaerobic digestate contains >90% water, the high nutrient content of digestate makes it economically and technically intractable to treatment by existing wastewater treatment technologies. This study separately assessed the feasibility of nutrient removal from digestate by Rhizopus delemar DSM 905 and a culture of phosphate-accumulating organisms (PAOs). With Rhizopus delemar DSM 905, we investigated concomitant nutrient removal from digestate-supplemented medium and fumaric acid production, as a potentially economical strategy for digestate treatment. Following the cultivation of R. delemar DSM 905 in a fermentation medium containing 25% (v/v) digestate, the concentrations of Al, Cr, Cu, Fe, K, Mg, Mn, Pb, and Zn reduced 40, 12, 74, 96, 12, 26, 23%, ~18, and 28%, respectively. Similarly, the concentrations of total phosphorus, total nitrogen, phosphate (PO4-P), ammonium (NH4-N), nitrate (NO3-N), and sulfur decreased 93, 88, 97, 98, 69, and 13%, respectively. Concomitantly, cultures supplemented with 25 and 15% (v/v) digestate produced comparable titers of fumarate (~11 and ~ 17 g/L, respectively) to the digestate un-supplemented control cultures. With PAOs, we assessed the removal of total phosphorus, total nitrogen, PO4-P, and NH4-N, of which the concentrations reduced 86, 90%, ~99, and 100%, respectively in 60% (v/v) digestate. This study provides additional bases for microbial removal of excess nutrients from anaerobic digestate, with the potential to engender future water recovery from this waste stream that is currently largely recalcitrant to treatment.

Whole-Genome Sequence and Fermentation Characteristics of Enterobacter hormaechei UW0SKVC1: A Promising Candidate for Detoxification of Lignocellulosic Biomass Hydrolysates and Production of Value-Added Chemicals.

Enterobacter hormaechei is part of the Enterobacter cloacae complex (ECC), which is widespread in nature. It is a facultative Gram-negative bacterium of medical and industrial importance. We assessed the metabolic and genetic repertoires of a new Enterobacter isolate. Here, we report the whole-genome sequence of a furfural- and 5-hydroxymethyl furfural (HMF)-tolerant strain of E. hormaechei (UW0SKVC1), which uses glucose, glycerol, xylose, lactose and arabinose as sole carbon sources. This strain exhibits high tolerance to furfural (IC50 = 34.2 mM; ~3.3 g/L) relative to Escherichia coli DH5α (IC50 = 26.0 mM; ~2.5 g/L). Furfural and HMF are predominantly converted to their less-toxic alcohols. E. hormaechei UW0SKVC1 produces 2,3-butanediol, acetoin, and acetol, among other compounds of industrial importance. E. hormaechei UW0SKVC1 produces as high as ~42 g/L 2,3-butanediol on 60 g/L glucose or lactose. The assembled genome consists of a 4,833,490-bp chromosome, with a GC content of 55.35%. Annotation of the assembled genome revealed 4586 coding sequences and 4516 protein-coding genes (average length 937-bp) involved in central metabolism, energy generation, biodegradation of xenobiotic compounds, production of assorted organic compounds, and drug resistance. E. hormaechei UW0SKVC1 shows considerable promise as a biocatalyst and a genetic repository of genes whose protein products may be harnessed for the efficient bioconversion of lignocellulosic biomass, abundant glycerol and lactose-replete whey permeate to value-added chemicals.

Development of an in vitro genotoxicity assay to detect retroviral vector-induced lymphoid insertional mutants.

Safety assessment in retroviral vector-mediated gene therapy remains challenging. In clinical trials for different blood and immune disorders, insertional mutagenesis led to myeloid and lymphoid leukemia. We previously developed the In Vitro Immortalization Assay (IVIM) and Surrogate Assay for Genotoxicity Assessment (SAGA) for pre-clinical genotoxicity prediction of integrating vectors. Murine hematopoietic stem and progenitor cells (mHSPCs) transduced with mutagenic vectors acquire a proliferation advantage under limiting dilution (IVIM) and activate stem cell- and cancer-related transcriptional programs (SAGA). However, both assays present an intrinsic myeloid bias due to culture conditions. To detect lymphoid mutants, we differentiated mHSPCs to mature T cells and analyzed their phenotype, insertion site pattern, and gene expression changes after transduction with retroviral vectors. Mutagenic vectors induced a block in differentiation at an early progenitor stage (double-negative 2) compared to fully differentiated untransduced mock cultures. Arrested samples harbored high-risk insertions close to Lmo2, frequently observed in clinical trials with severe adverse events. Lymphoid insertional mutants displayed a unique gene expression signature identified by SAGA. The gene expression-based highly sensitive molecular readout will broaden our understanding of vector-induced oncogenicity and help in pre-clinical prediction of retroviral genotoxicity.

A pro B cell population forms the apex of the leukemic hierarchy in Hoxa9/Meis1-dependent AML.

Relapse is a major challenge to therapeutic success in acute myeloid leukemia (AML) and can be partly associated with heterogeneous leukemic stem cell (LSC) properties. In the murine Hoxa9/Meis1-dependent (H9M) AML model, LSC potential lies in three defined immunophenotypes, including Lin-cKit+ progenitor cells (Lin-), Gr1+CD11b+cKit+ myeloid cells, and lymphoid cells (Lym+). Previous reports demonstrated their interconversion and distinct drug sensitivities. In contrast, we here show that H9M AML is hierarchically organized. We, therefore, tracked the developmental potential of LSC phenotypes. This unexpectedly revealed a substantial fraction of Lin- LSCs that failed to regenerate Lym+ LSCs, and that harbored reduced leukemogenic potential. However, Lin- LSCs capable of producing Lym+ LSCs as well as Lym+ LSCs triggered rapid disease development suggestive of their high relapse-driving potential. Transcriptional analyses revealed that B lymphoid master regulators, including Sox4 and Bach2, correlated with Lym+ LSC development and presumably aggressive disease. Lentiviral overexpression of Sox4 and Bach2 induced dedifferentiation of H9M cells towards a lineage-negative state in vitro as the first step of lineage conversion. This work suggests that the potency to initiate a partial B lymphoid primed transcriptional program as present in infant AML correlates with aggressive disease and governs the H9M LSC hierarchy.

Latent Medical Conditions of Drivers Involved in Road Traffic Collisions in Ghana: Implication from Autopsy Findings.

BACKGROUND: Unlike some neighboring countries like Nigeria, few studies on actual causes, impact and the prevention of road traffic collisions have been carried out in Ghana. There is the need for further research and this study sought to link injuries that caused the death of drivers involved in vehicular collisions to the latent possible contributing diseases and medical conditions in these drivers and how these conditions predispose them to the collisions. METHODS: This is a retrospective study that used the forensic autopsy records of driver fatalities and various injuries and medical conditions of drivers involved in road traffic collisions. Information on all drivers was retrieved from archives at the Komfo Anokye Teaching Hospital' Pathology Unit. Demographics and cause of death were used in selecting the cases, including all driver-related road traffic collisions from 2009 to 2014. RESULTS: A total of 1842 road traffic collisions were recorded with 127 of them being driver related. There were 31 cases (24.4%) in 2014; the highest cases recorded for this study, with the least recorded in 2009 with only 12 cases (9.4%). There were 121 (95.3%) male drivers and 6 female drivers (4.7%). Most of the male drivers were between the ages of 30 and 39 with 39 cases, while that of females was between 40 and 49 years with 6 cases. There was no significant statistical correlation between age and sex (P = .124). No statistical correlation also existed between sex and year (P = .331). Pathologies of all body systems were established. Cardiovascular diseases were the most prevalent systemic medical condition seen in the drivers with 44.1%. CONCLUSION: The study established that the drivers had various latent medical conditions and all these could lead to possible incapacitation, affecting driver judgment, leading to collisions on the road. The National Road Safety Commission (NRSC) should request medical screening before issuing driver licenses.

Detecting Ureaplasma urealyticum among HIV-infected women with or without human papillomavirus using real-time PCR with the ANYPLEX™ II STI-7 assay system.

OBJECTIVES: Human immunodeficiency virus (HIV) infections increase the chances of women contracting human papillomavirus (HPV), the prime causative agent of cervical cancer. Additionally, Ureaplasma urealyticum is the most frequent pathogen prevailing in sexually transmitted diseases in HIV-infected women. The aim of this study was to determine the presence of U. urealyticum among HIV-infected women with or without HPV in Ghana. METHODS: DNA samples were extracted from cervico-vaginal swabs obtained from 96 HIV-infected women attending the Kumasi South Hospital. U. urealyticum in the DNA samples was detected by real-time PCR with the Anyplex™ II STI-7 detection assay. Microsoft Excel was used to analyze the data, and the Chi-square test was used to determine associations and dependence among the variables. RESULTS: Among the study population, 93.75% (90/96) were positive for at least one pathogen.In total, 36.5% (35/96) of the women were infected by U. urealyticum, with 30.21% (29/96) being co-infected with HPV. There was no significant association (95% CI, p > 0.05) between U. urealyticum and HPV status among the HIV-infected women. CONCLUSION: U. urealyticum and HPV are highly pathogenic, and their prevalence in this study reiterates the need for their routine screening in HIV-infected patients.