Assessing Adverse Health Effects of Long-Term Exposure to Low Levels of Ambient Air Pollution: Implementation of Causal Inference Methods.

This report provides a final summary of the principal findings and key conclusions of a study supported by an HEI grant aimed at "Assessing Adverse Health Effects of Long-Term Exposure to Low Levels of Ambient Air Pollution." It is the second and final report on this topic. The study was designed to advance four critical areas of inquiry and methods development. First, it focused on predicting short- and long-term exposures to ambient fine particulate matter (PM2.5), nitrogen dioxide (NO2), and ozone (O3) at high spatial resolution (1 km × 1 km) for the continental United States over the period 2000-2016 and linking these predictions to health data. Second, it developed new causal inference methods for estimating exposure-response (ER) curves (ERCs) and adjusting for measured confounders. Third, it applied these methods to claims data from Medicare and Medicaid beneficiaries to estimate health effects associated with short- and long-term exposure to low levels of ambient air pollution. Finally, it developed pipelines for reproducible research, including approaches for data sharing, record linkage, and statistical software. Our HEI-funded work has supported an extensive portfolio of analyses and the development of statistical methods that can be used to robustly understand the health effects of short- and long-term exposure to low levels of ambient air pollution. Our Phase 1 report (Dominici et al. 2019) provided a high-level overview of our statistical methods, data analysis, and key findings, grouped into the following five areas: (1) exposure prediction, (2) epidemiological studies of ambient exposures to air pollution at low levels, (3) sensitivity analysis, (4) methodological contributions in causal inference, and (5) an open access research data platform. The current, final report includes a comprehensive overview of the entire research project.Considering our (1) massive study population, (2) numerous sensitivity analyses, and (3) transparent assessment of covariate balance indicating the quality of causal inference for simulating randomized experiments, we conclude that conditionally on the required assumptions for causal inference, our results collectively indicate that long-term PM2.5 exposure is likely to be causally related to mortality. This conclusion assumes that the causal inference assumptions hold and, more specifically, that we accounted adequately for confounding bias. We explored various modeling approaches, conducted extensive sensitivity analyses, and found that our results were robust across approaches and models. This work relied on publicly available data, and we have provided code that allows for reproducibility of our analyses.Our work provides comprehensive evidence of associations between exposures to PM2.5, NO2, and O3 and various health outcomes. In the current report, we report more specific results on the causal link between long-term exposure to PM2.5 and mortality, even at PM2.5 levels below or equal to 12 µg/m3, and mortality among Medicare beneficiaries (ages 65 and older). This work relies on newly developed causal inference methods for continuous exposure.For the period 2000-2016, we found that all statistical approaches led to consistent results: a 10-µg/m3 decrease in PM2.5 led to a statistically significant decrease in mortality rate ranging between 6% and 7% (= 1 - 1/hazard ratio [HR]) (HR estimates 1.06 [95% CI, 1.05 to 1.08] to 1.08 [95% CI, 1.07 to 1.09]). The estimated HRs were larger when studying the cohort of Medicare beneficiaries that were always exposed to PM2.5 levels lower than 12 µg/m3 (1.23 [95% CI, 1.18 to 1.28] to 1.37 [95% CI, 1.34 to 1.40]).Comparing the results from multiple and single pollutant models, we found that adjusting for the other two pollutants slightly attenuated the causal effects of PM2.5 and slightly elevated the causal effects of NO2 exposure on all-cause mortality. The results for O3 remained almost unchanged.We found evidence of a harmful causal relationship between mortality and long-term PM2.5 exposures adjusted for NO2 and O3 across the range of annual averages between 2.77 and 17.16 µg/m3 (included >98% of observations) in the entire cohort of Medicare beneficiaries across the continental United States from 2000 to 2016. Our results are consistent with recent epidemiological studies reporting a strong association between long-term exposure to PM2.5 and adverse health outcomes at low exposure levels. Importantly, the curve was almost linear at exposure levels lower than the current national standards, indicating aggravated harmful effects at exposure levels even below these standards.There is, in general, a harmful causal impact of long-term NO2 exposures to mortality adjusted for PM2.5 and O3 across the range of annual averages between 3.4 and 80 ppb (included >98% of observations). Yet within low levels (annual mean ≤53 ppb) below the current national standards, the causal impacts of NO2 exposures on all-cause mortality are nonlinear with statistical uncertainty.The ERCs of long-term O3 exposures on all-cause mortality adjusted for PM2.5 and NO2 are almost flat below 45 ppb, which shows no statistically significant effect. Yet we observed an increased hazard when the O3 exposures were higher than 45 ppb, and the HR was approximately 1.10 when comparing Medicare beneficiaries with annual mean O3 exposures of 50 ppb versus those with 30 ppb.institutions, including those that support the Health Effects Institute; therefore, it may not reflect the views or policies of these parties, and no endorsement by them should be inferred.A list of abbreviations and other terms appears at the end of this volume.

Assessing Adverse Health Effects of Long-Term Exposure to Low Levels of Ambient Air Pollution: Phase 1.

INTRODUCTION: This report provides a summary of major findings and key conclusions supported by a Health Effects Institute grant aimed at "Assessing Adverse Health Effects of Long-Term Exposure to Low Levels of Ambient Pollution." Our study was designed to advance four critical areas of inquiry and methods development. METHODS: First, our work focused on predicting short- and long-term exposures to ambient PM2.5 mass (particulate matter ≤ 2.5µm in aerodynamic diameter) and ozone (O3) at high spatial resolution (1 km × 1 km) for the continental United States during the period 2000-2012 and linking these predictions to health data. Second, we developed new causal inference methods for exposure-response (ER) that account for exposure error and adjust for measured confounders. We applied these methods to data from the New England region. Third, we applied standard regression methods using Medicare claims data to estimate health effects that are associated with short- and long-term exposure to low levels of ambient air pollution. We conducted sensitivity analyses to assess potential confounding bias due to lack of extensive information on behavioral risk factors in the Medicare population using the Medicare Current Beneficiary Survey (MCBS) (nationally representative sample of approximately 15,000 Medicare enrollees per year), which includes abundant data on individual-level risk factors including smoking. Finally, we have begun developing tools for reproducible research - including approaches for data sharing, record linkage, and statistical software. RESULTS: Our HEI-funded work has supported an extensive portfolio of analysis and the development of statistical methods that can be used to robustly understand the health effects of long- and short-term exposure to low levels of ambient air pollution. This report provides a high-level overview of statistical methods, data analysis, and key findings, as grouped into the following four areas: (1) Exposure assessment and data access; (2) Epidemiological studies of ambient exposures to air pollution at low levels; (3) Methodological contributions in causal inference; and (4) Open science research data platform. CONCLUSION: Our body of work, advanced by HEI, lends extensive evidence that short- and long-term exposure to PM2.5 and O3 is harmful to human health, increasing the risks of hospitalization and death, even at levels that are well below the National Ambient Air Quality Standards (NAAQS).

Preservation of root cementum: a comparative evaluation of power-driven versus hand instruments.

OBJECTIVES: The purpose of this study was to evaluate the effects of three distinct periodontal treatment methods in comparison with hand instrumentation on residual cementum of periodontal diseased teeth. Cementum can influence the activities of periodontal cells and may play an important regulatory role in periodontal treatment. The ideal method for periodontal therapy involves removal of biofilm, calculus and endotoxin while preserving root cementum. MATERIAL AND METHODS: Forty-eight caries free, single-rooted teeth in patients diagnosed with severe chronic periodontitis were treated using four different methods prior to extraction. The teeth were instrumented subgingivally at one approximal site either by hand curettes (HC), piezoelectric ultrasonic scalers (U), piezoelectric ultrasonic scalers following air polishing (U + AP) or air polishing (AP) alone. Following extraction of teeth, instrumented and non-instrumented sites were analysed with a dissecting microscope and SEM for measurement of the amount of and surface characteristics of residual cementum. RESULTS: The percentage of coronal cementum remaining following subgingival instrumentation was 84% for U, 80% for U + AP, 94% for AP and 65% for HC. Although subgingival instrumentation of apical portions of the cementum demonstrated 6% less retained cementum in comparison with coronal portions, the amount of retained cementum with AP was still significantly greater than with HC. SEM results found the smoothest root surfaces were produced by the HC followed by the AP, while root surfaces instrumented by U or U + AP presented grooves and scratches. CONCLUSIONS: This study demonstrated that AP was superior to U devices in preserving cementum, whereas HC were the most effective instruments in removing cementum.

A preliminary comparison of the effect of 0.3% versus 0.2% chlorhexidine mouth rinse on de novo plaque formation: a monocentre randomized double-blind crossover trial.

OBJECTIVE: Chlorhexidine (CHX) is considered the gold standard against gram-negative microorganisms. Little has been written about the simultaneous influence that both time and concentration could have on antiplaque formation effectiveness of CHX. The aim of this study is to compare the clinical and microbiological effectiveness of two different CHX concentrations and time applications in a 4-day plaque regrowth study model. MATERIAL AND METHODS: Twenty volunteers were enrolled in a randomized double-blind crossover study comparing the effectiveness of CHX 0.3% and CHX 0.2% mouth rinses applied for 15 and 30 s, respectively. Plaque index (PII), total bacterial counts and the detection of specific periopathogens were measured at the 5th day of each mouth rinse application. Taste acceptance was evaluated using a questionnaire. RESULTS: Chlorhexidine 0.3% resulted in a statistically greater reductions (10%) in PIl and periopathogens compared to CHX 0.2%. Furthermore, patients reported comparable taste acceptance in both groups. CONCLUSION: Chlorhexidine is an effective oral antiseptic. The CHX 0.3% mouth rinse formulation used for 15 s resulted in superior clinical and microbiological outcomes compared to the CHX 0.2% formulation, used for 30 s.

Health disparities among older adults following tropical cyclone exposure in Florida.

Tropical cyclones (TCs) pose a significant threat to human health, and research is needed to identify high-risk subpopulations. We investigated whether hospitalization risks from TCs in Florida (FL), United States, varied across individuals and communities. We modeled the associations between all storms in FL from 1999 to 2016 and over 3.5 million Medicare hospitalizations for respiratory (RD) and cardiovascular disease (CVD). We estimated the relative risk (RR), comparing hospitalizations during TC-periods (2 days before to 7 days after) to matched non-TC-periods. We then separately modeled the associations in relation to individual and community characteristics. TCs were associated with elevated risk of RD hospitalizations (RR: 4.37, 95% CI: 3.08, 6.19), but not CVD (RR: 1.04, 95% CI: 0.87, 1.24). There was limited evidence of modification by individual characteristics (age, sex, or Medicaid eligibility); however, risks were elevated in communities with higher poverty or lower homeownership (for CVD hospitalizations) and in denser or more urban communities (for RD hospitalizations). More research is needed to understand the potential mechanisms and causal pathways that might account for the observed differences in the association between tropical cyclones and hospitalizations across communities.

Morphological observations and Fractological considerations on orthodontics miniscrews.

AIM: The purpose of this study was to evaluate morphological and biomechanical characteristics of a commercial mini-implant. METHODS: Orthodontic low head mini implants, in the two available sizes of 1.5 and 2 mm, have been used. The electronic scanning microscope evaluation has been tested on 3 groups each one composed of 4 samples, observed in numerous enlargements and in the three dimensions of the space. The groups were composed of new mini implants, undamaged mini implants observed after their clinical use and mini-implants broken at the moment of their insertion or removal after their therapeutical use. RESULTS: The microscope analysis of unused mini implants has shown how the diameter reduction of the neck represents the area of greater weakness; such parameter is very important in determining screw resistance to fracture. In fact the fracture observed in vivo always happened to this degree. Some micro carvings and cracks were shown on the surface, these irregularities could facilitate or promote the enlargement of the cracking itself. CONCLUSION: From the results obtained it is clear that the steel is adequate for permanence in the oral cavity, and moreover the material itself is adapted to bear the pressure put on it during therapy. However, it remains advisable to make certain changes to better the surface and geometry of the miniscrews.

Air pollution and COVID-19 mortality in the United States: Strengths and limitations of an ecological regression analysis.

Assessing whether long-term exposure to air pollution increases the severity of COVID-19 health outcomes, including death, is an important public health objective. Limitations in COVID-19 data availability and quality remain obstacles to conducting conclusive studies on this topic. At present, publicly available COVID-19 outcome data for representative populations are available only as area-level counts. Therefore, studies of long-term exposure to air pollution and COVID-19 outcomes using these data must use an ecological regression analysis, which precludes controlling for individual-level COVID-19 risk factors. We describe these challenges in the context of one of the first preliminary investigations of this question in the United States, where we found that higher historical PM2.5 exposures are positively associated with higher county-level COVID-19 mortality rates after accounting for many area-level confounders. Motivated by this study, we lay the groundwork for future research on this important topic, describe the challenges, and outline promising directions and opportunities.

Evaluating the impact of long-term exposure to fine particulate matter on mortality among the elderly.

Many studies link long-term fine particle (PM2.5) exposure to mortality, even at levels below current U.S. air quality standards (12 micrograms per cubic meter). These findings have been disputed with claims that the use of traditional statistical approaches does not guarantee causality. Leveraging 16 years of data-68.5 million Medicare enrollees-we provide strong evidence of the causal link between long-term PM2.5 exposure and mortality under a set of causal inference assumptions. Using five distinct approaches, we found that a decrease in PM2.5 (by 10 micrograms per cubic meter) leads to a statistically significant 6 to 7% decrease in mortality risk. Based on these models, lowering the air quality standard to 10 micrograms per cubic meter would save 143,257 lives (95% confidence interval, 115,581 to 170,645) in one decade. Our study provides the most comprehensive evidence to date of the link between long-term PM2.5 exposure and mortality, even at levels below current standards.

Bayesian distributed lag models: estimating effects of particulate matter air pollution on daily mortality.

A distributed lag model (DLagM) is a regression model that includes lagged exposure variables as covariates; its corresponding distributed lag (DL) function describes the relationship between the lag and the coefficient of the lagged exposure variable. DLagMs have recently been used in environmental epidemiology for quantifying the cumulative effects of weather and air pollution on mortality and morbidity. Standard methods for formulating DLagMs include unconstrained, polynomial, and penalized spline DLagMs. These methods may fail to take full advantage of prior information about the shape of the DL function for environmental exposures, or for any other exposure with effects that are believed to smoothly approach zero as lag increases, and are therefore at risk of producing suboptimal estimates. In this article, we propose a Bayesian DLagM (BDLagM) that incorporates prior knowledge about the shape of the DL function and also allows the degree of smoothness of the DL function to be estimated from the data. We apply our BDLagM to its motivating data from the National Morbidity, Mortality, and Air Pollution Study to estimate the short-term health effects of particulate matter air pollution on mortality from 1987 to 2000 for Chicago, Illinois. In a simulation study, we compare our Bayesian approach with alternative methods that use unconstrained, polynomial, and penalized spline DLagMs. We also illustrate the connection between BDLagMs and penalized spline DLagMs. Software for fitting BDLagM models and the data used in this article are available online.

Multiple arm lipomatosis and posterior interosseus nerve palsy.

BACKGROUND: Lipomas are common benign soft tissue tumours which tend to be indolent and risk free. Lipomas rarely spread in the deep soft tissue causing posterior interosseous nerve (PIN) neuropathy. METHODS (CASE DESCRIPTION): We present two patients with multiple lipomatosis of the arms and PIN paralysis, with a brief review of the cases reported in literature. RESULTS AND CONCLUSION: We emphasize the role of electromyographic study as unique methodical capable to reveal an early radial nerve damage, permitting an optimal post-surgical nerve function recovering.

Changes in the recruitment curve of the soleus H-reflex associated with chronic low back pain.

OBJECTIVE: We investigated whether patients with chronic low back pain (CLBP) manifest changes in the excitability of the soleus H-reflex. METHODS: H-reflex stimulus-response curve was studied in 14 CLBP patients and 14 age-matched healthy subjects. H-threshold, H-maximum size, H-steepness and H-latency were determined for both legs. Homosynaptic depression (HD), following a train of H-reflexes, and presynaptic inhibition (PI) from flexor afferents onto soleus Ia afferents were also evaluated. RESULTS: H-threshold was significantly increased, H-size as a function of stimulus intensity was significantly different, and H-recruitment curve steepness was significantly lower in CLBP patients compared to healthy subjects. No significant difference in the amount of HD and PI of the H-reflex was found between the two groups. H-latency and Hmax/Mmax ratio was comparable between the subjects groups. CONCLUSIONS: In CLBP there is a reduced excitability of group Ia afferent fibres from the soleus muscle to which presynaptic factors do not seem to contribute and that presumably depend on changes in the peripheral sensory input. SIGNIFICANCE: Changes in H-reflex excitability may underlie a decrease in the gain of a peripheral sensor in CLBP. Estimation of soleus H-threshold and H-recruitment curve may contribute to the diagnostic evaluation of CLBP and may be used to monitor the efficacy of treatment.

Evidence of altered motor axon properties of the ulnar nerve in carpal tunnel syndrome.

OBJECTIVE: To analyse recruitment properties of ulnar nerve motor axons in 60 CTS patients with negative ulnar nerve electrodiagnostic tests. METHODS: Recruitment properties of the ulnar nerve were studied by analysing the relationship between the intensity of electrical stimulation and the size of motor response, i.e. the stimulus-response curve. Parameters of the curve (threshold, slope and plateau) were compared with those of the corresponding curve of the median nerve and both with parameters of 30 control curves. RESULTS: The ulnar nerve stimulus-response curve was strikingly abnormal and, except for severity, closely resembled that of the median nerve. The slope of the curve was significantly less than that of controls and decreased with increasing abnormalities of the median nerve. This suggested that the pathological process involving the ulnar nerve was contingent with the severity of median nerve involvement. CONCLUSIONS: We propose that the ulnar nerve may be subject to compression in Guyon's canal as a consequence of high pressure in the carpal tunnel of CTS patients. SIGNIFICANCE: Ectopic activity from ulnar axons may contribute to clinical spread of symptoms outside the median nerve territory in CTS. This does not exclude possible involvement of central plasticity mechanisms in producing extra-median symptoms in CTS patients.

[Smoke habit in a population of apprentices in middle of Italy]. / Abitudine al fumo in una popolazione di apprendisti dell'Italia centrale.

This study assessed the smoking habit in 7,159 apprentices (females: 53.3%) in central Italy. In the period 1996-2006, each apprentice underwent a compulsory health examination to assess fitness for work carried out by specialists in Occupational Medicine. Demographic data and information on schooling, smoking habit, alcohol and coffee intake were collected by a questionnaire. Overall, 51.9% of apprentices were smokers. Smokers were fewer among female apprentices than among the males (48.3% vs 55.9%; chi-square test, p < 0.001). No decrease in the prevalence of smokers was evident during the study period. Low educational level, and daily intake of alcohol and coffee significantly increased the risk of being a smoker. Finding a high percentage of smokers among Italian apprentices with a low educational level who are employed in jobs for which few qualifications are required indicates the need for work-place activities against tobacco smoking habit. Specialists in Occupational Medicine who perform regular health surveillance of workers could play a role in the anti-smoking campaign by integrating their routine work with brief counselling sessions aimed at preventing smoking in young workers and helping smokers to stop and by collaborating in work-place health promotion programmes.

Chronic blockade of the AT2 receptor with PD123319 impairs insulin signaling in C57BL/6 mice.

The renin-angiotensin system modulates insulin action. Angiotensin type 1 receptor exerts a deleterious effects while the angiotensin type 2 receptor (AT2R) appears to have beneficial effects providing protection against insulin resistance and type 2 diabetes. Although recent reports indicate that agonism of AT2R ameliorates diabetes and insulin resistance, the phenotype of AT2R-knockout mice seems to be controversial relating this aspect. Thus, in this study we have explored the role of AT2R in the control of insulin action. To that end, C57Bl/6 mice were administered the synthetic AT2R antagonist PD123319 for 21days (10mg/kg/day ip); vehicle treated animals were used as control. Glucose tolerance, metabolic parameters, in vivo insulin signaling in main insulin-target tissues as well as levels of adiponectin, TNF-α, MCP-1 and IL-6 in adipose tissue were assessed. AT2R blockade with PD123319 induced a marginal effect on glucose homeostasis but an important reduction in the insulin-induced phosphorylation of the insulin receptor and Akt in both liver and adipose tissue. Insulin signaling in skeletal muscle remained unaltered after treatment with PD123319, which could explain the minimal effect on glucose homeostasis induced by PD123319. Our current results reinforce the notion that the AT2R has a physiological role in the conservation of insulin action.

Changes in motor axon recruitment in the median nerve in mild carpal tunnel syndrome.

OBJECTIVE: To determine whether patients with mild carpal tunnel syndrome (CTS) and conventional electrodiagnostic evidence of selective involvement of sensory conduction show changes in motor axon recruitment in the median nerve. METHODS: Wrist-to-abductor pollicis brevis (APB) motor axon conduction was studied by analysing the relationship between the intensity of electrical stimulation and the size of motor response (input-output curve) in 30 CTS patients with conventional electrodiagnostic evidence of selective involvement of sensory conduction. Parameters (threshold, slope and plateau) of input-output curves were compared with those obtained in 30 controls. RESULTS: The slope of the input-output curve of CTS patients was less steep than that of controls. For stimulus intensity above M-wave threshold (MTh), fewer motor axons were recruited in patients than controls. CONCLUSIONS: Motor fibres are affected in CTS when conventional electrodiagnostic tests show normal motor conduction. Altered recruitment of motor axons could mainly be due to impairment of energy-dependent processes which affect temporal dispersion of the compound volley or axonal conduction block. SIGNIFICANCE: In mild CTS, motor fibres are more often affected than was originally thought. The sensitivity of wrist-to-APB motor conduction studies may be increased by using submaximal stimulus intensities.

Effect of cellulose and lignin on disintegration, antimicrobial and antioxidant properties of PLA active films.

This study reports the effects on antimicrobial, antioxidant, migration and disintegrability activities of ternary nanocomposite films based on poly(lactic acid) incorporating two biobased nanofillers, (cellulose nanocrystals (CNC) and lignin nanoparticles (LNP)), in two different amounts (1 and 3% wt.). Results from antimicrobial tests revealed a capacity to inhibit the Gram negative bacterial growth of Xanthomonas axonopodis pv. vesicatoria and Xanthomonas arboricola pv. pruni along the time, offering innovative opportunities against dangerous bacterial plant pathogens. LNP proved to be highly efficient in antioxidation activity, based on the disappearance of the absorption band at 517nm of the free radical, 2,2-diphenyl-1-picrylhydrazyl (DPPH) upon reduction by an antiradical compound; moreover the combination of LNP and CNC generates a synergistic positive effect in the antioxidation response of PLA ternary films. Furthermore, all the studied formulations showed a disintegrability value up to 90% after 15days of incubation in composting conditions. Migration results showed that the films can be considered suitable for application in food packaging field.

A measurement error model for time-series studies of air pollution and mortality.

One barrier to interpreting the observational evidence concerning the adverse health effects of air pollution for public policy purposes is the measurement error inherent in estimates of exposure based on ambient pollutant monitors. Exposure assessment studies have shown that data from monitors at central sites may not adequately represent personal exposure. Thus, the exposure error resulting from using centrally measured data as a surrogate for personal exposure can potentially lead to a bias in estimates of the health effects of air pollution. This paper develops a multi-stage Poisson regression model for evaluating the effects of exposure measurement error on estimates of effects of particulate air pollution on mortality in time-series studies. To implement the model, we have used five validation data sets on personal exposure to PM10. Our goal is to combine data on the associations between ambient concentrations of particulate matter and mortality for a specific location, with the validation data on the association between ambient and personal concentrations of particulate matter at the locations where data have been collected. We use these data in a model to estimate the relative risk of mortality associated with estimated personal-exposure concentrations and make a comparison with the risk of mortality estimated with measurements of ambient concentration alone. We apply this method to data comprising daily mortality counts, ambient concentrations of PM10measured at a central site, and temperature for Baltimore, Maryland from 1987 to 1994. We have selected our home city of Baltimore to illustrate the method; the measurement error correction model is general and can be applied to other appropriate locations.Our approach uses a combination of: (1) a generalized additive model with log link and Poisson error for the mortality-personal-exposure association; (2) a multi-stage linear model to estimate the variability across the five validation data sets in the personal-ambient-exposure association; (3) data augmentation methods to address the uncertainty resulting from the missing personal exposure time series in Baltimore. In the Poisson regression model, we account for smooth seasonal and annual trends in mortality using smoothing splines. Taking into account the heterogeneity across locations in the personal-ambient-exposure relationship, we quantify the degree to which the exposure measurement error biases the results toward the null hypothesis of no effect, and estimate the loss of precision in the estimated health effects due to indirectly estimating personal exposures from ambient measurements.

Increased sensitivity to GH in liver of Ames dwarf (Prop1df/Prop1df) mice related to diminished CIS abundance.

To investigate the influence of chronic GH deficiency on GH signaling in vivo, we have analyzed Janus kinase (JAK) 2/signal transducers and activators of transcription (STAT) 5 GH signaling pathway, and its regulation by the suppressors of the cytokine signaling SOCS and by the JAK2-interacting protein SH2-Bbeta, in liver of Ames dwarf (Prop1df/Prop1df) mice, which are severely deficient in GH, prolactin and TSH, and of their normal littermates. Prop1df/Prop1df mice displayed unaltered GH receptor, JAK2 and STAT5a/b protein levels. No significant differences in the basal tyrosine-phosphorylation levels of JAK2 and STAT5a/b were found between both groups of animals. After in vivo administration of a high GH dose (5 microg/g body weight (BW)), the tyrosine-phosphorylation levels of JAK2 and STAT5a/b increased significantly, reaching similar values in normal and dwarf mice. However, after stimulation with lower GH doses (50 and 15 ng/g BW) the tyrosine-phosphorylation level of STAT5a/b was higher in dwarf mice. The protein content of CIS, a SOCS protein that inhibits STAT5 signaling, was approximately 80% lower in dwarf mice liver, while SOCS-2 and SOCS-3 levels were unaltered. The content of SH2-Bbeta, a modulator of JAK2 activity, was reduced by approximately 30% in dwarf mice, although this was associated with normal JAK2 response to a high GH dose. In summary, Prop1df/Prop1df mice display increased hepatic sensitivity to GH, an effect that could be related to the lower abundance of CIS in this tissue. Furthermore, the lower CIS content found in this model of GH deficiency suggests that CIS protein levels are regulated by GH in vivo.

Nitrosative Stress and Apoptosis by Intravenous Ferumoxytol, Iron Isomaltoside 1000, Iron Dextran, Iron Sucrose, and Ferric Carboxymaltose in a Nonclinical Model.

Iron is involved in the formation as well as in the scavenging of reactive oxygen and nitrogen species. Thus, iron can induce as well as inhibit both oxidative and nitrosative stress. It also has a key role in reactive oxygen and nitrogen species-mediated apoptosis. We assessed the differences in tyrosine nitration and caspase 3 expression in the liver, heart, and kidneys of rats treated weekly with intravenous ferumoxytol, iron isomaltoside 1000, iron dextran, iron sucrose and ferric carboxymaltose (40 mg iron/kg body weight) for 5 weeks. Nitrotyrosine was quantified in tissue homogenates by Western blotting and the distribution of nitrotyrosine and caspase 3 was assessed in tissue sections by immunohistochemistry. Ferric carboxymaltose and iron sucrose administration did not result in detectable levels of nitrotyrosine or significant levels of caspase 3 vs. control in any of the tissue studied. Nitrotyrosine and caspase 3 levels were significantly (p<0.01) increased in all assessed organs of animals treated with iron dextran and iron isomaltoside 1000, as well as in the liver and kidneys of ferumoxytol-treated animals compared to isotonic saline solution (control). Nitrotyrosine and caspase 3 levels were shown to correlate positively with the amount of Prussian blue-detectable iron(III) deposits in iron dextran- and iron isomaltoside 1000-treated rats but not in ferumoxytol-treated rats, suggesting that iron dextran, iron isomaltoside 1000 and ferumoxytol induce nitrosative (and oxidative) stress as well as apoptosis via different mechanism(s).

Bionanocomposite films based on plasticized PLA-PHB/cellulose nanocrystal blends.

Optically transparent plasticized poly(lactic acid) (PLA) based bionanocomposite films intended for food packaging were prepared by melt blending. Materials were plasticized with 15wt% of acetyl(tributyl citrate) (ATBC) to improve the material processability and to obtain flexibile films. Poly(hydroxybutyrate) (PHB) was used to increase PLA crystallinity. The thermal stability of the PLA-PHB blends was improved by the addition of 5 wt% of cellulose nanocrystals (CNC) or modified cellulose nanocrystals (CNCs) synthesized from microcrystalline cellulose. The combination of ATBC and cellulose nanocrystals, mainly the better dispersed CNCs, improved the interaction between PLA and PHB. Thus, an improvement on the oxygen barrier and stretchability was achieved in PLA-PHB-CNCs-ATBC which also displayed somewhat UV light blocking effect. All bionanocomposite films presented appropriate disintegration in compost suggesting their possible applications as biodegradable packaging materials.