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The article discusses the study which examines the differences in storm water quality measured during sampling techniques at several locations in stream cross-sections. The researchers have observed different degrees of concentration variability within stream cross-sections for dissolved and particulate constituents. The study indicates some substantial temporal concentration variability for dissolved and particulate constituents, but fails to account for potential variability in cross-sectional concentrations.

Improving access to care, health outcomes, and patient satisfaction are primary objectives for healthcare practices. This article outlines benefits, concerns, and possible challenges of shared medical appointments (SMAs) for patients and providers. The SMA model was designed to support providers' demanding schedules by allowing patients with the same chronic condition to be seen in a group setting. By concentrating on patient education and disease management, interactive meetings provide an opportunity for patients to share both successes and struggles with others experiencing similar challenges. Studies demonstrated that SMAs improved patient access, enhanced outcomes, and promoted patient satisfaction. This article describes the potential benefits of SMAs for patients with chronic heart disease, which consumes a large number of healthcare dollars related to hospital admissions, acute exacerbations, and symptom management Education for self-management of chronic disease can become repetitive and time consuming. The SMA model introduces a fresh and unique of healthcare visits, allowing providers to devote more time and attention to patients and improve productivity. The SMA model provides an outstanding method for nurse practitioners to demonstrate their role as a primary care provider, by leading patients in group discussions and evaluating their current health status. Patient selection, preparation, and facilitation of an SMA are discussed to demonstrate the complementary nature of an SMA approach in a healthcare practice. [ABSTRACT FROM AUTHOR]

The article focuses on neutralization potential (NP) determination of siderite (FeCO3) with the use of selected oxidants. The determination of NP can be interfered by siderite. Neutrolize acid has an effect on siderite . The reaction of the siderite has an effect on the NP. An examination was conducted on the effects of hydrogen peroxide, oxygen, and potassium permanganate on the NP of siderate samples. Oxidation treatments on siderite samples have different results. Oxidation with oxygen is the most accurate in examining NP.

On-farm adoption of agricultural conservation practices or management alternatives depends on conservation ethic, social pressure, regulatory attention, and perceived impact on yield and economic return. Although agro-economic and environmental impacts are assumed to conflict, little research has been conducted to address potential tradeoffs and provide a scientific basis for decision-making. Thus, this 16-year evaluation of adaptive nutrient management was conducted in the Texas Blackland Prairies ecoregion on six fields with structural conservation practices already in place. Each field was randomly selected to receive either commercial fertilizer or poultry litter at rates of 4.5 to 13.4 Mg ha-1. Two major nutrient management adaptations were made (i.e., soil test nitrogen [N] rate recommendations in 2009, and reduction of fallow period length and cover cropping during prolonged fallow periods in 2013). Important results included (1) soil test recommendations that consider historical crop yields reduced N application 25% to 38% for low rates of poultry litter, but did not reduce profits; (2) interannual variability of economic and weather conditions contributed to the lack of statistically significant differences in profit, although profit reduction for high nutrient rate treatments was clear; and (3) litter application, especially at rates in excess of crop phosphorus (P) needs, also increased runoff P losses by 1 to 1.4 kg ha-1 indicating the need for careful management of organic nutrient sources. Results of this long-term study showed that maintaining or increasing economic return does not have to be sacrificed to improve environmental impacts, which is an important consideration as producers make on-farm management decisions. [ABSTRACT FROM AUTHOR]

Water quality experts have suggested that no-till induces phosphorus (P) stratification, which may exacerbate soluble P (SP) runoff from agricultural fields, contributing to eutrophication. Conservationists have been concerned about increased SP loading to Lake Erie, which has been partially blamed on adoption of no-till and the concomitant P stratification of no-till soils. This study was conducted to provide better insight into the potential link between P stratification from no-till soils and P losses via runoff with the objective of exploring P fertilization strategies on P stratification and P runoff from a corn (Zea mays L.)–soybean (Glycine max L.) rotation. Plots were established with nine treatments, including unfertilized, diammonium phosphate (DAP) applications, monoammonium phosphate (MAP) applications, surface applied, injecting fertilizer or tilling fertilizer in, and the use of cover crops. Fertilizer applications were made at 24.4 kg P ha−1 (21.8 lb P ac−1) every other year or at 9.6 kg P ha−1 (8.7 lb P ac−1) every year. Disking, which was intended to minimize P stratification, resulted in the greatest stratification, with significantly higher water SP and Mehlich 3 P in the 0 to 5 cm (0 to 2 in) soil layer compared to the other treatments. There were no differences in SP or total P (TP) runoff from rainfall simulations between fertilizer source (MAP versus DAP) or fertilizer rate (annual versus biennial). The highest SP concentrations observed were from DAP applied to cover crops at the high application rate (24.4 kg P ha−1 applied every other year). This may suggest cover crops are not the ideal practice to decrease SP losses from agricultural fields. Incorporation of fertilizer reduced SP but increased erosion and could potentially increase TP loss. Injecting liquid fertilizer (polyphosphate [Poly]) at the time of planting resulted in lower SP and TP loads than surface applied fertilizers. We encourage other researchers to confirm these results at the field-to-watershed scale to ensure there are not unintended consequences of adopting this fertilization strategy. Further, fertilizer dealers, crop consultants, and farmers should be encouraged to consider liquid fertilizer applications as one option to minimize P losses. [ABSTRACT FROM AUTHOR]

Application of litter and other organic by-products to agricultural land off site of animal production facilities has created both environmental concerns and agro-economic opportunities, but limited long-term, field-scale data are available to guide management decisions. Thus, the objective of this study was to determine the water quality effects of repeated annual poultry litter application as a cropland and pasture fertilizer. Eight years of data collected on ten field-scale watersheds indicated several significant water quality differences based on litter rate (0.0 to 13.4 Mg ha-1 [0 to 6 ton ac-1]) and land use (cropland and pasture). On cropland fields, increasing litter rates (with corresponding decreases in supplemental inorganic nitrogen [Ni) increased runoff orthophosphate phosphorus (PO4-P) concentrations but reduced extreme high nitrate nitrogen (NO3-N) concentrations. Whereas run off PO4-P concentrations were somewhat similar between land uses, NO3-N concentrations were much lower in pasture runoff because of supplemental inorganic N application, reduced nutrient uptake potential, and faster litter mineralization on cropland. Although considerable variability was observed, intra-annual runoff NO3-N and PO4-P concentrations generally exhibited curvilinear decay based on time since fertilizer application. In spite of repeated annual litter application and buildup of soil phosphorus (P) at high litter rates, few long-term trends in N and P runoff were evident due to the dynamic interaction between transport and source factors. These results support several practical implications, specifically: (1) combining organic and inorganic nutrient sources can be environmentally friendly and economically sound if application rates are carefully managed; (2) high runoff N and P concentrations can occur from well-managed fields, which presents difficulty in regulating edge-of-field water quality; and (3) change in the animal industry mindset to view by-products as marketable resources could mitigate environmental problems, provide alternative fertilizer sources, and enhance animal industry revenue opportunities. [ABSTRACT FROM AUTHOR]

The size of the harmful algal blooms in western Lake Erie is strongly correlated with agricultural phosphorus (P) loading from tributaries. Despite farmers' efforts to reduce sediment-bound P loadings and fertilize using current guidance, the media and public have singled them out as the culprit in Lake Erie re-eutrophication. In this paper, two farmer surveys were used to evaluate if farmers in the Lake Erie region follow P fertilizer recommendations, and we also review historic and current P management guidance provided by the scientific community and agricultural industry. The majority (56% to 80%) of farmers apply P fertilizers at or below the current fertility recommendations. Wholesale agronomic changes (e.g., no-tillage adoption, crop cultivar advances, and fertilizer application and formulation) have occurred since current fertilizer recommendations were developed. Although crop P uptake mechanisms have not changed, these agronomic changes have altered P cycling in soil and water. Based on these results, it is time that the scientific community and agricultural industry acknowledge that our current guidance may be contributing to eutrophication. We must ask whether or not we have (1) developed appropriate fertility guidance, (2) developed and recommended appropriate practices to protect water quality, (3) adequately considered "the law of unintended consequences" in conservation recommendations, and (4) focused too much on short-term economic outcomes while disregarding environmental quality. Improved understanding, reconsideration of traditional recommendations, and wider farmer adoption of the most effective practices are needed to develop a sustainable agricultural system in the Western Lake Erie Basin that produces needed commodities while preserving ecosystem integrity. [ABSTRACT FROM AUTHOR]

Excess nutrient loading from numerous sources (e.g., agricultural and urban runoff, treatment plant discharge, and streambank erosion) continues to adversely impact water resources, and determination of the cause(s) of accelerated nutrient enrichment has become a contentious and litigious issue in several US regions. This paper addresses one fundamental question: What are acceptable levels of nutrients in runoff from agricultural fields? It focuses on the field scale where farmers and ranchers make management decisions. Not answering this question limits the effectiveness of on-farm management and policy alternatives to address agriculture's contribution. To answer the question, some might suggest "direct comparison"with reference site data, existing criteria/standards, or measured data compilations. Alternatively, "indirect assessments"using soil test phosphorus (P) levels, P indices, field-scale models, or certainty programs might be suggested. Thus, to provide a scientific basis for policy debate and management decisions related to nutrient runoff from agricultural fields, we evaluated "direct comparisons"with measured data from case studies and evaluated "indirect assessment"alternatives. While acknowledging that scientific challenges and practical realities exist for each alternative, we concluded that certainty programs offer the most promise for ensuring acceptable nutrient runoff, and that field-scale models linked with watershed decision support tools are the most promising for assessing impacts on downstream water quality. Recognizing the reality that some nutrient loss is unavoidable from natural and anthropogenic sources, agriculture, industry, and municipalities are each encouraged to commit to implementing enhanced management where needed to minimize their sector's contribution to excess nutrients in our nation's waters. [ABSTRACT FROM AUTHOR]

The application of poultry litter to agricultural land has become a topic of interest for policy makers due to public concern about its effects on water quality. The Soil and Water Assessment Tool (SWAT) version 2005 is designed to assess nonpoint and point sources of pollution. In this study, six subwatersheds in Texas (HUC-8; 12070101) are used to evaluate the model's ability to simulate water quality at a small scale. Each of these subwatersheds randomly received poultry litter rates of 0.0 to 13.4 Mg ha-1. Monthly and daily data from 2002 were used for calibration purposes, while 2000, 2001, 2003, and 2004 were used for validation. The SCS runoff curve number for moisture condition II (CN2) and the soil evaporation compensation factor (ESCO) parameters were found to be more sensitive than the surface runoff lag time (SURLAG) and initial soil water content expressed as a fraction of field capacity (FFCB). The monthly and daily runoff model simulations for the six subwatersheds resulted in calibration Nash-Sutcliffe efficiency (NSE) values of 0.59 and 0.53 and validation NSE values 0.82 and 0.80, respectively. The monthly and daily R² runoff values for the six subwatersheds resulted in calibration values of at least 0.60 and 0.53 and validation R² values of 0.86 and 0.81, respectively. The observed trends included SWAT's overestimation of runoff in the dry periods and underestimation in the wet periods. The monthly NSE and R² values for sediment and nutrient losses were generally above 0.4 and 0.5, respectively. Paired t-tests for the monthly manually adjusted parameter simulation of sediment, organic N and P, NO3-IV, and soluble P for the 2000-2004 period losses showed that their respective SWAT means were not significantly different from the measured values (α = 0.05), except for NO3-N losses for the Y10 subwatershed (p-value 0.042). The control subwatershed's measured and simulated water quality results were significantly different (α = 0.05) from the treated subwatersheds, most likely due to the amount of inorganic N present. Almost all of the subwatersheds that had poultry litter applied resulted in higher sediment, organic N, organic P, and soluble P losses than the control subwatershed upon averaging thee monthly validation values. High NO3-N losses may have been a function of poultry litter and commercial fertilizers being applied before a large rainfall event occurred. The subwatersheds that received smaller amounts of commercial fertilizer and/or poultry litter lost more sediment, organic N, and organic P than the subwatersheds that received the higher litter and/or fertilizer treatments. Overall, the SWAT simulated the hydrology and the water quality constituents at the subwatershed scale more adequately when all of the data were used to simulate the model, as evidenced by statistical measures. [ABSTRACT FROM AUTHOR]

The scientific community has not established an adequate understanding of the uncertainty inherent in measured water quality data, which is introduced by four procedural categories: streamflow measurement, sample collection, sample preservation/storage, and laboratory analysis. Although previous research has produced valuable information on relative differences in procedures within these categories, little information is available that compares the procedural categories or presents the cumulative uncertainty in resulting water quality data. As a result, quality control emphasis is often misdirected, and data uncertainty is typically either ignored or accounted for with an arbitrary margin of safety. Faced with tile need for scientifically defensible estimates of data uncertainty to support water resource management, the objectives of this research were to: (1) compile selected published information on uncertainty related to measured stream, flow and water quality data for small watersheds, (2) use a root mean square error propagation method to compare the uncertainty introduced by each procedural category, and (3) use the error propagation method to determine the cumulative probable uncertainty in measured streamflow, sediment, and nutrient data. Best case, typical, and worst case "data quality" scenarios were examined. Averaged across all constituents, the calculated cumulative probable uncertainty (±%) contributed under typical scenarios ranged from 6% to 19% for streamflow measurement, from 4% to 48% for sample collection, from 2% to 16% for sample preservation/storage, and from 5% to 21% for laboratory analysis. Under typical conditions, errors in storm loads ranged from 8% to 104% for dissolved nutrients, from 8% to 110% for total N and P, and from 7% to 53% for TSS. Results indicated that uncertainty can increase substantially under poor measurement conditions and limited quality control effort. This research provides introductory scientific estimates of uncertainty in measured water quality data. The results and procedures presented should also assist modelers in quantifying tile "quality" of calibration and evaluation data sets, determining model accuracy goals, and evaluating model performance. [ABSTRACT FROM AUTHOR]

Phosphorus (P) indices were developed to address nonpoint source P losses from agricultural fields; however, only limited information on P index performance at the field- and watershed-scale is available. Evaluation of P indices is necessary to provide the basis of modification and improvement of their usefulness as P management tools. In this study, the ability of the Texas P index to estimate P loss potential was evaluated by comparison with measured annual P loads over three years on four new pasture and six new cultivated litter application sites in the Texas Blackland Prairie. The Arkansas and Iowa P indices were also evaluated. The Texas and Iowa versions were able to provide reasonable estimates of P loss potential as illustrated with significant linear relationships (p <0.01) between P index values and measured annual P loads. In general, the P index values, Mehlich 3 soil test P, and poultry litter application rate were better correlated with dissolved P concentrations and loads (r² ranged from 0.12 to 0.91) than with total P and particulate P loads (r² ranged from 0.00-0.31). A major source of error in P index load estimations was their inability to capture variability in annual soil erosion. This source of error was dramatically reduced by using measured erosion instead of estimated annual average erosion (average r² values increased from 0.24 to 0.58). While these results illustrate a potential for the P Indices to make relative P loss assessments, research on incentives to prevent buildup of soil P levels, linkages between P levels in soils and receiving waters, and other important issues related to the use of P indices is warranted. [ABSTRACT FROM AUTHOR]

Lenacapavir (LEN) is a highly potent, long-acting antiretroviral medication for treating people infected with muti-drug-resistant HIV-1 phenotypes. The inhibitor targets multifaceted functions of the viral capsid protein (CA) during HIV-1 replication. Previous studies have mainly focused on elucidating LEN's mode of action during viral ingress. Additionally, the inhibitor has been shown to interfere with mature capsid assembly during viral egress. However, the mechanism for how LEN affects HIV-1 maturation is unknown. Here, we show that pharmacologically relevant LEN concentrations do not impair proteolytic processing of Gag in virions. Instead, we have elucidated the primary mechanism for highly potent inhibition of HIV-1 maturation by sub-stoichiometric LEN:CA ratios. The inhibitor exerts opposing effects on formation of CA pentamers versus hexamers, the key capsomere intermediates in mature capsid assembly. LEN impairs formation of pentamers, whereas it induces assembly of hexameric lattices by imposing an opened CA conformation and stabilizing a dimeric form of CA. Consequently, LEN treatment results in morphologically atypical virus particles containing malformed, hyper-stable CA assemblies, which fail to infect target cells. Moreover, we have uncovered an inverse correlation between inhibitor potency and CA levels in cell culture assays, which accounts for LEN's ability to potently (with pM EC 50 values) inhibit HIV-1 maturation at clinically relevant drug concentrations., Author Summary: Lenacapavir (LEN) is the first-in-class HIV-1 capsid targeting antiretroviral that exhibits multimodal modality to inhibit both early and late steps of viral replication. Our studies here have elucidated previously undescribed structural and mechanistic bases for a highly potent antiviral activity of LEN during viral egress. These findings will inform clinical applications of LEN as a potent HIV-1 maturation inhibitor and aid the development of second-generation inhibitors targeting assembly of the mature viral capsid.

Lenacapavir (LEN) is the first in class viral capsid protein (CA) targeting antiretroviral for treating multi-drug-resistant HIV-1 infection. Clinical trials and cell culture experiments have identified resistance associated mutations (RAMs) in the vicinity of the hydrophobic CA pocket targeted by LEN. The M66I substitution conferred by far the highest level of resistance to the inhibitor compared to other RAMs. Here we investigated structural and mechanistic bases for how the M66I change affects LEN binding to CA and viral replication. The high-resolution X-ray structure of the CA(M66I) hexamer revealed that the β-branched side chain of Ile66 induces steric hindrance specifically to LEN thereby markedly reducing the inhibitor binding affinity. By contrast, the M66I substitution did not affect binding of Phe-Gly (FG)-motif-containing cellular cofactors CPSF6, NUP153, or SEC24C, which engage the same hydrophobic pocket of CA. In cell culture the M66I variant did not acquire compensatory mutations or replicate in the presence of LEN. Analysis of viral replication intermediates revealed that HIV-1 (M66I CA) predominantly formed correctly matured viral cores, which were more stable than their wildtype counterparts. The mutant cores stably bound to the nuclear envelope but failed to penetrate inside the nucleus. Furthermore, the M66I substitution markedly altered HIV-1 integration targeting. Taken together, our findings elucidate mechanistic insights for how the M66I change confers remarkable resistance to LEN and affects HIV-1 replication. Moreover, our structural findings provide powerful means for future medicinal chemistry efforts to rationally develop second generation inhibitors with a higher barrier to resistance., Importance: Lenacapavir (LEN) is a highly potent and long-acting antiretroviral that works by a unique mechanism of targeting the viral capsid protein. The inhibitor is used in combination with other antiretrovirals to treat multi-drug-resistant HIV-1 infection in heavily treatment-experienced adults. Furthermore, LEN is in clinical trials for preexposure prophylaxis (PrEP) with interim results indicating 100 % efficacy to prevent HIV-1 infections. However, one notable shortcoming is a relatively low barrier of viral resistance to LEN. Clinical trials and cell culture experiments identified emergent resistance mutations near the inhibitor binding site on capsid. The M66I variant was the most prevalent capsid substitution identified in patients receiving LEN to treat muti-drug resistant HIV-1 infections. The studies described here elucidate the underlying mechanism by which the M66I substitution confers a marked resistance to the inhibitor. Furthermore, our structural findings will aid future efforts to develop the next generation of capsid inhibitors with enhanced barriers to resistance.

Allosteric HIV-1 integrase (IN) inhibitors (ALLINIs) are investigational antiretroviral agents that potently impair virion maturation by inducing hyper-multimerization of IN and inhibiting its interaction with viral genomic RNA. The pyrrolopyridine-based ALLINI pirmitegravir (PIR) has recently advanced into phase 2a clinical trials. Previous cell culture-based viral breakthrough assays identified the HIV-1 (Y99H/A128T IN) variant that confers substantial resistance to this inhibitor. Here, we have elucidated the unexpected mechanism of viral resistance to PIR. Although both Tyr99 and Ala128 are positioned within the inhibitor binding V-shaped cavity at the IN catalytic core domain (CCD) dimer interface, the Y99H/A128T IN mutations did not substantially affect the direct binding of PIR to the CCD dimer or functional oligomerization of full-length IN. Instead, the drug-resistant mutations introduced a steric hindrance at the inhibitor-mediated interface between CCD and C-terminal domain (CTD) and compromised CTD binding to the CCD Y99H/A128T + PIR complex. Consequently, full-length IN Y99H/A128T was substantially less susceptible to the PIR-induced hyper-multimerization than the WT protein, and HIV-1 (Y99H/A128T IN) conferred >150-fold resistance to the inhibitor compared with the WT virus. By rationally modifying PIR, we have developed its analog EKC110, which readily induced hyper-multimerization of IN Y99H/A128T in vitro and was ~14-fold more potent against HIV-1 (Y99H/A128T IN) than the parent inhibitor. These findings suggest a path for developing improved PIR chemotypes with a higher barrier to resistance for their potential clinical use.IMPORTANCEAntiretroviral therapies save the lives of millions of people living with HIV (PLWH). However, the evolution of multi-drug-resistant viral phenotypes is a major clinical problem, and there are limited or no treatment options for heavily treatment-experienced PLWH. Allosteric HIV-1 integrase inhibitors (ALLINIs) are a novel class of antiretroviral compounds that work by a unique mechanism of binding to the non-catalytic site on the viral protein and inducing aberrant integrase multimerization. Accordingly, ALLINIs potently inhibit both wild-type HIV-1 and all drug-resistant viral phenotypes that have so far emerged against currently used therapies. Pirmitegravir, a highly potent and safe investigational ALLINI, is currently advancing through clinical trials. Here, we have elucidated the structural and mechanistic bases behind the emergence of HIV-1 integrase mutations in infected cells that confer resistance to pirmitegravir. In turn, our findings allowed us to rationally develop an improved ALLINI with substantially enhanced potency against the pirmitegravir-resistant virus., Competing Interests: Kyungjin Kim was the chief executive officer of ST Pharm Co. Ltd. No other authors declare a potential conflict of interest.

The HIV-1 integrase (IN) plays a critical role in the viral lifecycle by integrating the viral DNA into the host chromosome. The catalytic function of IN has been exploited as a target, with five drugs acting as active site binders (IN strand transfer inhibitors, INSTIs). However, IN mutations conferring low-level resistance to INSTIs have been reported. Therefore, new IN inhibitors with different mechanisms of action are needed. The allosteric inhibition of IN, exerted by allosteric IN inhibitors (ALLINIs), is gaining interest. ALLINIs inhibit IN by inducing aberrant IN multimerization with different mechanisms. Furthermore, recent discoveries unveiled that IN has an under-studied yet equally vital second function. This involves IN binding to the RNA genome in virions, necessary for proper virion maturation. In this work, we describe a series of quinolinonyl derivatives as inhibitors of both the IN catalytic functions and IN-RNA interactions, which impair both early and late steps of viral replication., Competing Interests: The authors declare no competing financial interest., (© 2024 American Chemical Society.)

Lenacapavir (LEN) is a long-acting, highly potent HIV-1 capsid (CA) inhibitor. The evolution of viral variants under the genetic pressure of LEN identified Q67H, N74D, and Q67H/N74D CA substitutions as the main resistance associated mutations (RAMs). Here, we determined high-resolution structures of CA hexamers containing these RAMs in the absence and presence of LEN. Our findings reveal that the Q67H change induces a conformational switch, which adversely affects the inhibitor binding. In the unliganded protein, the His67 side chain adopts the closed conformation by projecting into the inhibitor binding pocket and thereby creating steric hindrance with respect to LEN. Upon the inhibitor binding, the His67 side chain repositions to the open conformation that closely resembles the Gln67 side chain in the WT protein. We propose that the switch from the closed conformation to the open conformation, which is needed to accommodate LEN, accounts for the reduced inhibitor potency with respect to the Q67H CA variant. The N74D CA change results in the loss of a direct hydrogen bond and in induced electrostatic repulsions between CA and LEN. The double Q67H/N74D substitutions exhibited cumulative effects of respective single amino acid changes. An examination of LEN binding kinetics to CA hexamers revealed that Q67H and N74D CA changes adversely influenced the inhibitor binding affinity ( K D ) by primarily affecting the dissociation rate constant ( k off ). We used these structural and mechanistic findings to rationally modify LEN. The resulting analog exhibited increased potency against the Q67H/N74D viral variant. Thus, our studies provide a means for the development of second-generation inhibitors with enhanced barriers to resistance. IMPORTANCE LEN is an investigational long-acting agent for future HIV-1 treatment regimens. While ongoing clinical trials have highlighted a largely beneficial profile of LEN for the treatment of HIV-1 infected people with limited therapy options, one notable shortcoming is a relatively low barrier of viral resistance to the inhibitor. Cell culture-based viral breakthrough assays identified N74D, Q67H, and N74D/Q67H capsid changes as the main resistance associated mutations (RAMs). N74D and Q67H capsid substitutions have also emerged in clinical trials in some patients who received subcutaneous LEN. Understanding the structural basis behind viral resistance to LEN is expected to aid in the rational development of improved inhibitors with enhanced barriers to resistance. Here, we report high resolution structures of the main drug resistant capsid variants, which provide mechanistic insight into the viral resistance to LEN. We used these findings to develop an improved inhibitor, which exhibited enhanced activity against the viral Q67H/N74D capsid phenotype compared with that of parental LEN.

Cellular proteins CPSF6, NUP153 and SEC24C play crucial roles in HIV-1 infection. While weak interactions of short phenylalanine-glycine (FG) containing peptides with isolated capsid hexamers have been characterized, how these cellular factors functionally engage with biologically relevant mature HIV-1 capsid lattices is unknown. Here we show that prion-like low complexity regions (LCRs) enable avid CPSF6, NUP153 and SEC24C binding to capsid lattices. Structural studies revealed that multivalent CPSF6 assembly is mediated by LCR-LCR interactions, which are templated by binding of CPSF6 FG peptides to a subset of hydrophobic capsid pockets positioned along adjoining hexamers. In infected cells, avid CPSF6 LCR-mediated binding to HIV-1 cores is essential for functional virus-host interactions. The investigational drug lenacapavir accesses unoccupied hydrophobic pockets in the complex to potently impair HIV-1 inside the nucleus without displacing the tightly bound cellular cofactor from virus cores. These results establish previously undescribed mechanisms of virus-host interactions and antiviral action., (© 2022. The Author(s).)

Pantothenate (Pan) is an essential nutrient required by both the mosquito vector and malaria parasite. We previously demonstrated that increasing pantothenate kinase (PanK) activity and co-enzyme A (CoA) biosynthesis led to significantly decreased parasite infection prevalence and intensity in the malaria mosquito Anopheles stephensi. In this study, we demonstrate that Pan stores in A. stephensi are a limited resource and that manipulation of PanK levels or activity, via small molecule modulators of PanK or transgenic mosquitoes, leads to the conversion of Pan to CoA and an overall reduction in Pan levels with minimal to no effects on mosquito fitness. Transgenic A. stephensi lines with repressed insulin signaling due to PTEN overexpression or repressed c-Jun N-terminal kinase (JNK) signaling due to MAPK phosphatase 4 (MKP4) overexpression exhibited enhanced PanK levels and significant reductions in Pan relative to non-transgenic controls, with the PTEN line also exhibiting significantly increased CoA levels. Provisioning of the PTEN line with the small molecule PanK modulator PZ-2891 increased CoA levels while provisioning Compound 7 decreased CoA levels, affirming chemical manipulation of mosquito PanK. We assessed effects of these small molecules on A. stephensi lifespan, reproduction and metabolism under optimized laboratory conditions. PZ-2891 and Compound 7 had no impact on A. stephensi survival when delivered via bloodmeal throughout mosquito lifespan. Further, PZ-2891 provisioning had no impact on egg production over the first two reproductive cycles. Finally, PanK manipulation with small molecules was associated with minimal impacts on nutritional stores in A. stephensi mosquitoes under optimized rearing conditions. Together with our previous data demonstrating that PanK activation was associated with significantly increased A. stephensi resistance to Plasmodium falciparum infection, the studies herein demonstrate a lack of fitness costs of mosquito Pan depletion as a basis for a feasible, novel strategy to control parasite infection of anopheline mosquitoes., Competing Interests: Declaration of competing interest None., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: Many of the leading causes of infant mortality are diagnosed prenatally, presenting providers with the ability to present perinatal palliative care planning as an option., Objective: Our study adds to the literature both by describing infant interaction with the health care system and by gaining deeper understanding of the maternal experience after being offered perinatal palliative care., Methods: The study was conducted at a public university-based medical center in the Midwest. Phase 1 consisted of a retrospective review of electronic medical records of 27 mother-infant pairs offered perinatal palliative care, 18 of whom elected to develop a perinatal palliative care. Phase 2 consisted of a focus group and interviews of seven of the mothers., Results: In the initial phase of this study, results revealed differences regarding the infant's end-of-life trajectory, including location of death, number of invasive procedures, and death in the setting of withholding versus withdrawing life-sustaining treatment. Highlighting that without a perinatal palliative care plan in place, the default treatment for infants with prenatally diagnosed life-limiting conditions is likely to be invasive and painful with often times minimal likelihood of long-term survival. Analysis of interview and focus group data revealed three themes: care, choice, and legacy., Conclusion: The authors used their experience with the health care system to draw implications for practice from the focus group and interview data, which care can serve to promote women feeling cared for and cared about, as well as promote opportunities for hope during a fragile pregnancy.

Objective: The current study examined pain and neurogenic inflammation responses to topical capsaicin during the interictal period (between headache) and their relationship with plasma oxytocin in individuals with migraine., Background: Individuals with migraine can experience generalized (extracephalic) hyperalgesia, which can persist even between headache attacks. Elevated levels of plasma and cerebrospinal fluid oxytocin have been observed during migraine attacks, oxytocin levels being positively associated with the intensity of migraine symptoms. However, whether oxytocin plays a role in the mechanisms of generalized pain sensitization and neurogenic inflammation during the interictal period has not been studied yet. Understanding migraineurs' interictal pain phenotype and endogenous oxytocin might help identify individuals who would benefit from intranasal oxytocin treatment., Methods: Thirty-two subjects with migraine and 26 healthy controls underwent pain testing. The current study compared capsaicin-induced pain, central sensitization (areas of secondary mechanical allodynia and hyperalgesia), and neurogenic inflammation (capsaicin-induced flare) responses on the nondominant volar forearm between migraineurs and healthy controls. Additionally, we studied plasma oxytocin levels and their relationship to migraine symptoms, experimental pain and affect., Results: The results indicated a significant group effect (P = .019): Migraineurs reported greater capsaicin-induced pain unpleasantness (M = 1.2, SD = 1.4) on a 0-10 scale and showed larger areas of flare (LnM = 2.8, SD = 0.4) than healthy controls (M = 0.5, SD = 0.8; LnM = 2.6, SD = 0.4; ps < .032). In a subgroup analysis, enhanced capsaicin-induced pain unpleasantness was found in the chronic (P = .007), but not the episodic (Ps > .200), migraineurs. The oxytocin levels were elevated in migraineurs and accounted for 18% of the group difference in capsaicin-induced pain unpleasantness. Within migraineurs, interictal oxytocin levels were negatively associated with psychological distress (Ps < .030). However, during the interictal period, pain sensitivity in extracephalic regions and plasma oxytocin levels were unrelated to migraine symptom parameters (Ps > .074). Lastly, the results found no group difference in areas of secondary mechanical allodynia and hyperalgesia (Ps >.298)., Conclusion: The current study revealed that individuals with migraine exhibit enhanced extracephalic capsaicin-induced pain unpleasantness and flare responses during interictal periods. In addition, migraineurs, especially those with chronic migraine, had slightly elevated interictal oxytocin levels compared to controls, which was associated with their affective component of experimental pain. Therefore, treatment targeting affective pain during the interictal period may help to reduce generalized pain in migraine. Furthermore, endogenous increases in oxytocin may be a compensatory mechanism that may help decrease affective distress in migraineurs. The therapeutic effects of intranasal oxytocin may benefit migraineurs by reducing their affective distress., (© 2017 American Headache Society.)

Increasing evidence suggests thrombospondin-1 (TSP-1), a potent proatherogenic matricellular protein, as a putative link between hyperglycemia and atherosclerotic complications in diabetes. We previously reported that the micronutrient chromium picolinate (CrP), with long-standing cardiovascular benefits, inhibits TSP-1 expression in glucose-stimulated human aortic smooth muscle cells in vitro. Here, we investigated the atheroprotective action of orally administered CrP in type 1 diabetic apolipoprotein E-deficient (ApoE -/- ) mice and elucidated the role of TSP-1 in this process. CrP decreased lipid burden and neointimal thickness in aortic root lesions of hyperglycemic ApoE -/- mice; also, smooth muscle cell (SMC), macrophage and leukocyte abundance was prevented coupled with reduced cell proliferation. Attenuated lesion progression was accompanied with inhibition of hyperglycemia-induced TSP-1 expression and reduced protein O-glycosylation following CrP treatment; also, PCNA and vimentin (SMC synthetic marker) expression were reduced while SM-MHC (SMC contractile marker) levels were increased. To confirm a direct role of TSP-1 in diabetic atherosclerosis, hyperglycemic TSP-1 -/- /ApoE -/- double knockout mice were compared with age-matched hyperglycemic ApoE -/- littermates. Lack of TSP-1 prevented lesion formation in hyperglycemic ApoE -/- mice, mimicking the atheroprotective phenotype of CrP-treated mice. These results suggest that therapeutic TSP-1 inhibition may have important atheroprotective potential in diabetic vascular disease.

Multiple sclerosis (MS) is a complex disease influenced by genetic and environmental contributing factors. Endocrine disrupting compounds (EDCs) such as bisphenol A (BPA) affect gene expression and hormone-regulated systems throughout the body. We investigated the effects of BPA on Theiler's-virus induced demyelination (TVID), a mouse model of MS. Perinatal BPA exposure, combined with viral infection, resulted in a decreased level of viral antibodies, accelerated the onset of TVID symptoms, increased inflammation in both the spinal cord and digestive tract, and amplified immune-related gene expression changes induced by viral infection. These results demonstrate the effect of BPA on the trajectory of TVID, and illustrate how multiple factors collectively influence autoimmune disease.

The combined use of ZnO, Mg, MgO, and silk provides routes to classes of thin-film transistors and mechanical energy harvesters that are soluble in water and biofluids. Experimental and theoretical studies of the operational aspects and dissolution properties of this type of transient electronics technology illustrate its various capabilities. Application opportunities range from resorbable biomedical implants, to environmentally dissolvable sensors, and degradable consumer electronics., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Reversibility of aberrant methylation via pharmacological means is an attractive target for therapies through epigenetic reprogramming. To establish that pharmacologic reversal of methylation could result in functional inhibition of angiogenesis, we undertook in vitro and in vivo studies of thrombospondin-1 (TSP1), a known inhibitor of angiogenesis. TSP1 is methylated in several malignancies, and can inhibit angiogenesis in melanoma xenografts. We analyzed effects of 5-Aza-deoxycytidine (5-Aza-dC) on melanoma cells in vitro to confirm reversal of promoter hypermethylation and restoration of TSP1 expression. We then investigated the effects of TSP1 expression on new blood vessel formation and tumor growth in vivo. Finally, to determine potential for clinical translation, the methylation status of TSP1 promoter regions of nevi and melanoma tissues was investigated., Results: 5-Aza-dC reduced DNA (cytosine-5)-methyltransferase 1 (DNMT1) protein, reversed promoter hypermethylation, and restored TSP1 expression in five melanoma cell lines, while having no effect on TSP1 protein levels in normal human melanocytes. In in vivo neovascularization studies, mice were implanted with melanoma cells (A375) either untreated or treated with 5Aza-dC. Vessels at tumor sites were counted by an observer blinded to treatments and the number of tumor vessels was significantly decreased at pretreated tumor sites. This difference occurred before a significant difference in tumor volumes was seen, yet in further studies the average tumor volume in mice treated in vivo with 5-Aza-dC was decreased by 55% compared to untreated controls. Knockdown of TSP1 expression with shRNA enhanced tumor-induced angiogenesis by 68%. Analyses of promoter methylation status of TSP1 in tumors derived from untreated and treated mice identified 67% of tumors from untreated and 17% of tumors from treated mice with partial methylation consistent with the methylation specific PCR analysis of A375 cells. Examination of methylation patterns in the promoter of TSP1 and comparison of aberrantly methylated TSP1 in melanoma with non-malignant nevi identified a significantly higher frequency of promoter methylation in tumor samples from melanoma patients., Conclusions: Pharmacological reversal of methylation silenced TSP1 had functional biological consequences in enhancing angiogenesis inhibition and inducing antitumor effects to decrease murine melanoma growth. Angiogenesis inhibition is an additional mechanism by which epigenetic modulators can have antitumor effects., (Copyright © 2012 International Society of Matrix Biology. Published by Elsevier B.V. All rights reserved.)

Background: Regional genetic connectivity models are critical for successful conservation and management of marine species. Even though rocky shore invertebrates have been used as model systems to understand genetic structure in some marine environments, our understanding of connectivity in Caribbean communities is based overwhelmingly on studies of tropical fishes and corals. In this study, we investigate population connectivity and diversity of Cittarium pica, an abundant rocky shore trochid gastropod that is commercially harvested across its natural range, from the Bahamas to Venezuela., Methodology/principal Findings: We tested for genetic structure using DNA sequence variation at the mitochondrial COI and 16S loci, AMOVA and distance-based methods. We found substantial differentiation among Caribbean sites. Yet, genetic differentiation was associated only with larger geographic scales within the Caribbean, and the pattern of differentiation only partially matched previous assessments of Caribbean connectivity, including those based on larval dispersal from hydrodynamic models. For instance, the Bahamas, considered an independent region by previous hydrodynamic studies, showed strong association with Eastern Caribbean sites in our study. Further, Bonaire (located in the east and close to the meridional division of the Caribbean basin) seems to be isolated from other Eastern sites., Conclusions/significance: The significant genetic structure and observed in C. pica has some commonalities in pattern with more commonly sampled taxa, but presents features, such as the differentiation of Bonaire, that appear unique. Further, the level of differentiation, together with regional patterns of diversity, has important implications for the application of conservation and management strategies in this commercially harvested species.