Attacking Latent HIV with convertibleCAR-T Cells, a Highly Adaptable Killing Platform.

Current approaches to reducing the latent HIV reservoir entail first reactivating virus-containing cells to become visible to the immune system. A critical second step is killing these cells to reduce reservoir size. Endogenous cytotoxic T-lymphocytes (CTLs) may not be adequate because of cellular exhaustion and the evolution of CTL-resistant viruses. We have designed a universal CAR-T cell platform based on CTLs engineered to bind a variety of broadly neutralizing anti-HIV antibodies. We show that this platform, convertibleCAR-T cells, effectively kills HIV-infected, but not uninfected, CD4 T cells from blood, tonsil, or spleen and only when armed with anti-HIV antibodies. convertibleCAR-T cells also kill within 48 h more than half of the inducible reservoir found in blood of HIV-infected individuals on antiretroviral therapy. The modularity of convertibleCAR-T cell system, which allows multiplexing with several anti-HIV antibodies yielding greater breadth and control, makes it a promising tool for attacking the latent HIV reservoir.

Variations in Groundwater Level and Microtopography Influence Desert Plant Communities in Shallow Aquifer Areas.

An improved understanding of the relationships among vegetation, groundwater level, and microtopography is crucial for making well-informed management decisions in areas with shallow groundwater resources. We measured plant species abundance/composition and richness in relation to depth to groundwater (DTW) and microtopography in Owens Valley, California, particularly in areas where DTW ranged from 0 to 4 m. Sampling occurred along 67 vegetation transects across three community types. Relationships between DTW and community composition were evaluated using non-metric multidimensional scaling (NMS), while non-parametric multiplicative regression was used to relate DTW and microtopography to species abundance. The dominant gradient in species composition (NMS Axis 1) explained ~51% of variation in our distance matrix and was most strongly associated (r = 0.55) with DTW. The graminoids Juncus arcticus, Leymus triticoides, and Distichlis spicata had strong affinities toward areas with the shallowest DTW (<1.5 m). One salt-adapted species Sporobolus airoides and one shrub Ericameria nauseosa dominated areas with intermediate DTW (1.5-2.0 m), whereas the shrubs Atriplex torreyi, Sarcobatus vermiculatus, and Artemisia tridentata were dominant in areas with deeper DTW (>2.0 m). Variation in microtopography affected species abundance and increased species richness for vegetation communities at either extreme of the water table gradient, shallow, and deep DTW but not the intermediate DTW. Findings indicate that desert plant communities from shallow aquifers have adapted to different DTW and microtopography conditions and that considering those adaptations may be important to manage groundwater and vegetation resources in these areas.

Heterozygous deletion of Atbf1 by the Cre-loxP system in mice causes preweaning mortality.

ATBF1 is a large nuclear protein that contains multiple zinc-finger motifs and four homeodomains. In mammals, ATBF1 regulates differentiation, and its mutation and/or downregulation is involved in tumorigenesis in several organs. To gain more insight into the physiological functions of ATBF1, we generated and validated a conditional allele of mouse Atbf1 in which exons 7 and 8 were flanked by loxP sites (Atbf1(flox) ). Germline deletion of a single Atbf1 allele was achieved by breeding to EIIa-cre transgenic mice, and Atbf1 heterozygous mice displayed reduced body weight, preweaning mortality, increased cell proliferation, and attenuated cytokeratin 18 expression, indicating haploinsufficiency of Atbf1. Floxed Atbf1 mice will help us understand such biological processes as neuronal differentiation and tumorigenesis.

Synthesis and structural characterization of an unprecedented nonmetal cation polyborate salt containing two different "isolated" polyborate anions: [H2en]2[B4O5(OH)4][B7O9(OH)5]·3H2O (en = H2NCH2CH2NH2).

The nonmetal cation polyborate salt of stoichiometry [H(2)en](2)[B(11)O(18)(OH)]·7H(2)O is obtained from the reaction of 1,2-diaminoethane and boric acid (1:5 ratio) in H(2)O/MeOH. An X-ray crystallographic study of the product reveals that the polyborate moiety is composed of two isolated hydrated polyborate anions: [B(4)O(5)(OH)(4)](2-) and [B(7)O(9)(OH)(5)](2-). The structure is templated by the cations with the anions forming a supramolecular H-bonded network, augmented by additional H-bonds involving the waters of crystallization and the cations.

convertibleCARs: A chimeric antigen receptor system for flexible control of activity and antigen targeting.

We have developed a chimeric antigen receptor (CAR) platform that functions as a modular system to address limitations of traditional CAR therapies. An inert form of the human NKG2D extracellular domain (iNKG2D) was engineered as the ectodomain of the CAR to generate convertibleCARTM-T cells. These cells were specifically directed to kill antigen-expressing target cells only in the presence of an activating bispecific adapter comprised of an iNKG2D-exclusive ULBP2-based ligand fused to an antigen-targeting antibody (MicAbodyTM). Efficacy against Raji tumors in NSG mice was dependent upon doses of both a rituximab-based MicAbody and convertibleCAR-T cells. We have also demonstrated that the exclusive ligand-receptor partnering enabled the targeted delivery of a mutant form of IL-2 to selectively promote the expansion of convertibleCAR-T cells in vitro and in vivo. By altering the Fv domains of the MicAbody or the payload fused to the orthogonal ligand, convertibleCAR-T cells can be readily targeted or regulated.

Diagnostic stewardship of C. difficile testing: a quasi-experimental antimicrobial stewardship study.

OBJECTIVE: We evaluated whether a diagnostic stewardship initiative consisting of ASP preauthorization paired with education could reduce false-positive hospital-onset (HO) Clostridioides difficile infection (CDI). DESIGN: Single center, quasi-experimental study. SETTING: Tertiary academic medical center in Chicago, Illinois. PATIENTS: Adult inpatients were included in the intervention if they were admitted between October 1, 2016, and April 30, 2018, and were eligible for C. difficile preauthorization review. Patients admitted to the stem cell transplant (SCT) unit were not included in the intervention and were therefore considered a contemporaneous noninterventional control group. INTERVENTION: The intervention consisted of requiring prescriber attestation that diarrhea has met CDI clinical criteria, ASP preauthorization, and verbal clinician feedback. Data were compared 33 months before and 19 months after implementation. Facility-wide HO-CDI incidence rates (IR) per 10,000 patient days (PD) and standardized infection ratios (SIR) were extracted from hospital infection prevention reports. RESULTS: During the entire 52 month period, the mean facility-wide HO-CDI-IR was 7.8 per 10,000 PD and the SIR was 0.9 overall. The mean ± SD HO-CDI-IR (8.5 ± 2.0 vs 6.5 ± 2.3; P < .001) and SIR (0.97 ± 0.23 vs 0.78 ± 0.26; P = .015) decreased from baseline during the intervention. Segmented regression models identified significant decreases in HO-CDI-IR (Pstep = .06; Ptrend = .008) and SIR (Pstep = .1; Ptrend = .017) trends concurrent with decreases in oral vancomycin (Pstep < .001; Ptrend < .001). HO-CDI-IR within a noninterventional control unit did not change (Pstep = .125; Ptrend = .115). CONCLUSIONS: A multidisciplinary, multifaceted intervention leveraging clinician education and feedback reduced the HO-CDI-IR and the SIR in select populations. Institutions may consider interventions like ours to reduce false-positive C. difficile NAAT tests.

Antibacterial efficacy of R-type pyocins towards Pseudomonas aeruginosa in a murine peritonitis model.

R-type pyocins are high-molecular-weight bacteriocins carried within the chromosomes of some bacterial species, such as Pseudomonas aeruginosa, and almost certainly evolved from lysogenic bacteriophages of the Myoviridae family. They contain no head structures and no DNA and are used as defense systems, usually against other strains of the same bacterial species. They bind with their tail fibers to targeted bacterial surface molecules and then kill by inserting a core or needle that dissipates the bacterial membrane potential. Their mechanism of action, high bactericidal potency (one pyocin particle can kill one bacterium), and focused spectrum suggest that R-type pyocins could be developed as antibacterial agents. In a lethal mouse peritonitis model, submicrogram quantities of pyocin prevent death from 90% lethal dose inocula of a pyocin-sensitive, clinical isolate of P. aeruginosa. We show here the dose response curves, treatment windows, or periods of response after infection and the several-log-unit acute reduction of bacterial load in blood and spleen samples, suggesting that R-type pyocins have several characteristics that one would expect from an effective therapeutic.

Retargeting R-type pyocins to generate novel bactericidal protein complexes.

R-type pyocins are high-molecular-weight bacteriocins that resemble bacteriophage tail structures and are produced by some Pseudomonas aeruginosa strains. R-type pyocins kill by dissipating the bacterial membrane potential after binding. The high-potency, single-hit bactericidal kinetics of R-type pyocins suggest that they could be effective antimicrobials. However, the limited antibacterial spectra of natural R-type pyocins would ultimately compromise their clinical utility. The spectra of these protein complexes are determined in large part by their tail fibers. By replacing the pyocin tail fibers with tail fibers of Pseudomonas phage PS17, we changed the bactericidal specificity of R2 pyocin particles to a different subset of P. aeruginosa strains, including some resistant to PS17 phage. We further extended this idea by fusing parts of R2 tail fibers with parts of tail fibers from phages that infect other bacteria, including Escherichia coli and Yersinia pestis, changing the killing spectrum of pyocins from P. aeruginosa to the bacterial genus, species, or strain that serves as a host for the donor phage. The assembly of active R-type pyocins requires chaperones specific for the C-terminal portion of the tail fiber. Natural and retargeted R-type pyocins exhibit narrow bactericidal spectra and thus can be expected to cause little collateral damage to the healthy microbiotae and not to promote the horizontal spread of multidrug resistance among bacteria. Engineered R-type pyocins may offer a novel alternative to traditional antibiotics in some infections.

An Evaluation of the Proposed Worker Protection Standard with Respect to Pesticide Exposure and Parkinson's Disease.

Citing a lack of information, the U.S. Environmental Protection Agency prudently did not account for the benefits of averting many chronic diseases in analyzing the Worker Protection Standards (WPS) revisions. We demonstrate that sufficient information can exist, using the example of the benefits to agricultural workers of reduced Parkinson's disease (PD) due to reduced pesticide exposure. We define the benefits as the monetary value gained by improving quality of lives of people who would otherwise develop PD, plus the value of medical care cost averted and income not lost due to being healthy. For estimation, we use readily available parameters and obtain odds ratios of developing PD by conducting a meta-analysis of studies linking pesticide exposure to PD. The sensitivity analysis varies the number of agricultural workers affected by the regulation, the probability of being diagnosed with PD, the measurement and the timing of the benefits. Our initial assessment is that the reduced PD benefits would be a small fraction of the total WPS revision costs. However, if we define benefits as the common environmental economics willingness to pay to avoid PD incidence, then they become a substantial fraction of the costs. Our analysis demonstrates that the benefits of averting PD from the WPS revisions can be estimated using existing information, and that the results are most sensitive to the choice of valuation of benefits to the worker. We encourage other researchers to extend our framework to other chronic ailments.

Nonsteroidal anti-inflammatory drug-activated gene-1 over expression in transgenic mice suppresses intestinal neoplasia.

BACKGROUND & AIMS: The nonsteroidal anti-inflammatory drug-activated gene (NAG-1) was identified as a proapoptotic, antitumorigenic protein in vitro, induced by many antitumorigenic and chemopreventive drugs including cyclooxygenase inhibitors. However, its antitumorigenic activity has not been elucidated in vivo. METHODS: Transgenic mice were generated that ubiquitously overexpress human NAG-1 under the control of a chicken beta-actin promoter (CAG). The NAG-1 transgenic mice (NAG-(Tg+)) were characterized, and then the antitumorigenic activity was evaluated with 2 colorectal carcinogenesis models: chemical induction with azoxymethane and genetic induction using the Apc(Min+) mutation. RESULTS: NAG-(Tg+) showed no apparent phenotype other than a reduction in body weight, particularly in males. To examine whether NAG-1 expression would suppress intestinal tumorigenesis, the NAG-(Tg+) mice were treated with the colorectal carcinogen azoxymethane. NAG-(Tg+) mice developed 50% fewer aberrant crypt foci and no tumors, in comparison with nontransgenic littermates. This result demonstrates that expression of this human protein in vivo can suppress chemically induced carcinogenesis in the colon. The NAG-(Tg+) mice were also crossed with Apc(Min+) mice to determine the effect of the transgene on intestinal polyp formation. NAG-(Tg+) mice heterozygous for the Apc(Min+) mutation had a significantly reduced polyp load (60%) compared with nontransgenic Apc(Min+) littermates. CONCLUSIONS: Our results support NAG-1 as an important regulator of intestinal adenoma growth in vivo and suggest that NAG-1 may act as a tumor suppressor gene.

Phreatophytic vegetation and groundwater fluctuations: a review of current research and application of ecosystem response modeling with an emphasis on great basin vegetation.

Although changes in depth to groundwater occur naturally, anthropogenic alterations may exacerbate these fluctuations and, thus, affect vegetation reliant on groundwater. These effects include changes in physiology, structure, and community dynamics, particularly in arid regions where groundwater can be an important water source for many plants. To properly manage ecosystems subject to changes in depth to groundwater, plant responses to both rising and falling groundwater tables must be understood. However, most research has focused exclusively on riparian ecosystems, ignoring regions where groundwater is available to a wider range of species. Here, we review responses of riparian and other species to changes in groundwater levels in arid environments. Although decreasing water tables often result in plant water stress and reduced live biomass, the converse is not necessarily true for rising water tables. Initially, rising water tables kill flooded roots because most species cannot tolerate the associated low oxygen levels. Thus, flooded plants can also experience water stress. Ultimately, individual species responses to either scenario depend on drought and flooding tolerance and the change in root system size and water uptake capacity. However, additional environmental and biological factors can play important roles in the severity of vegetation response to altered groundwater tables. Using the reviewed information, we created two conceptual models to highlight vegetation dynamics in areas with groundwater fluctuations. These models use flow charts to identify key vegetation and ecosystem properties and their responses to changes in groundwater tables to predict community responses. We then incorporated key concepts from these models into EDYS, a comprehensive ecosystem model, to highlight the potential complexity of predicting community change under different fluctuating groundwater scenarios. Such models provide a valuable tool for managing vegetation and groundwater use in areas where groundwater is important to both plants and humans, particularly in the context of climate change.

Advancing the theory and measurement of collective empowerment: a qualitative study.

Grounded Theory, a qualitative research method, was used to fully describe the construct of empowerment and its measurement in racially and ethnically diverse urban and rural neighborhoods. Forty-nine grass roots experts, primarily from six communities in Texas, participated via semi-structured interviews and focus groups. Observational data were also collected. Results revealed that the framework of collective empowerment has two main dimensions: ten processes necessary for developing greater empowerment and eight long-term outcomes that result from the process strategies. The two main dimensions of collective empowerment are cyclically, rather than linearly, related. While the results are unique to the communities studied because of the nature of the analysis, the process by which they were elucidated can be replicated in any setting. These findings suggest that the construct of collective empowerment can be both operationalized and evaluated at the community level.

Herpesvirus papio 2 encodes a virion host shutoff function.

Infection of baboons with herpesvirus papio 2 (HVP-2) produces a disease that is similar to human infection with herpes simplex viruses (HSV). Molecular characterization of HVP-2 has demonstrated that the virion contains a factor which rapidly shuts off host cell protein synthesis after infection. Reduction of host cell protein synthesis occurs in parallel with the degradation of mRNA species. A homolog of the HSV virion host shutoff (vhs) gene was identified by Southern and DNA sequence analysis. The sequence of the HVP-2 vhs gene homolog had greater than 70% identity with the vhs genes of HSV 1 and 2. Disruption of the HVP-2 vhs open reading frame diminished the ability of the virus to shut off protein synthesis and degrade cellular mRNA, indicating that this gene was responsible for the vhs activity. The HVP-2 model system provides the opportunity to study the biological role of vhs in the context of a natural primate host. Further development of this system will provide a platform for proof-of-concept studies that will test the efficacy of vaccines that utilize vhs-deficient viruses.

The genome of herpesvirus papio 2 is closely related to the genomes of human herpes simplex viruses.

Infection of baboons (Papio species) with herpesvirus papio 2 (HVP-2) produces a disease that is clinically similar to herpes simplex virus (HSV-1 and HSV-2) infection of humans. The development of a primate model of simplexvirus infection based on HVP-2 would provide a powerful resource to study virus biology and test vaccine strategies. In order to characterize the molecular biology of HVP-2 and justify further development of this model system we have constructed a physical map of the HVP-2 genome. The results of these studies have identified the presence of 26 reading frames that closely resemble HSV homologues. Furthermore, the HVP-2 genome shares a collinear arrangement with the genome of HSV. These studies further validate the development of the HVP-2 model as a surrogate system to study the biology of HSV infections.

A Limited Capacity Information Processing Interpretation of Conditioned Emotional Responding.

This investigation assessed the usefulness of a limited capacity information processing model for evaluation of stress. The assumption is that the task of anticipating an impending aversive event requires a portion of the limited capacity. Thus, such stress can be measured by a performance decrement in a concurrent task. Stress was manipulated in 14 male and female college students by means of a conditioned emotional response (CER) technique. Two tasks, motor tracking and counting backwards, were systematically studied as both primary and secondary tasks, depending on the presence or absence of reinforcement. The cognitive task of counting backwards was more sensitive to the processing demands imposed by stress than the motor task of tracking.