The M3 Muscarinic Acetylcholine Receptor Can Signal through Multiple G Protein Families.

The M3 muscarinic acetylcholine receptor (M3R) is a G protein-coupled receptor (GPCR) that regulates important physiologic processes, including vascular tone, bronchoconstriction, and insulin secretion. It is expressed on a wide variety of cell types, including pancreatic beta, smooth muscle, neuronal, and immune cells. Agonist binding to the M3R is thought to initiate intracellular signaling events primarily through the heterotrimeric G protein Gq. However, reports differ on the ability of M3R to couple to other G proteins beyond Gq. Using members from the four primary G protein families (Gq, Gi, Gs, and G13) in radioligand binding, GTP turnover experiments, and cellular signaling assays, including live cell G protein dissociation and second messenger assessment of cAMP and inositol trisphosphate, we show that other G protein families, particularly Gi and Gs, can also interact with the human M3R. We further show that these interactions are productive as assessed by amplification of classic second messenger signaling events. Our findings demonstrate that the M3R is more promiscuous with respect to G protein interactions than previously appreciated. SIGNIFICANCE STATEMENT: The study reveals that the human M3 muscarinic acetylcholine receptor (M3R), known for its pivotal roles in diverse physiological processes, not only activates intracellular signaling via Gq as previously known but also functionally interacts with other G protein families such as Gi and Gs, expanding our understanding of its versatility in mediating cellular responses. These findings signify a broader and more complex regulatory network governed by M3R and have implications for therapeutic targeting.

Luminescent lanthanide mixed N-phthaloylglycinate and terephthalate coordination polymers: Structural and optical properties.

A new class of lanthanide mixed-carboxylate ligands compounds with formula {[Ln2 (phthgly)4 (bdc)(H2 O)6 ]·(H2 O)4 }∞ , labelled as Ln3+ : Eu (1) and Gd (2) coordination polymers (CP) were synthesized under mild reaction conditions between lanthanide nitrate salts and a solution of N-phthaloylglycine (phthgly) and terephthalic (bdc) ligands. The (1) and (2) coordination polymers were formed by symmetric binuclear units, in which phthgly and bdc carboxylate ligands are coordinated to the lanthanide ions by different coordination modes. Surprisingly, all organic ligands participate in hydrogen bonding interactions, forming an extremally rigid crystalline structure. The red narrow emission bands from the 5 D0 →7 FJ transitions of the Eu3+ ion show a high colour purity. The intramolecular energy transfer process from L→Eu3+ ion has been discussed. The experimental intensity parameters (Ω2,4 ) reflect lower angular distortion and polarizability of the chemical environment around the metal ion compared with other Eu3+ compounds reported in the literature. This novel class of coordination polymer offers a more attractive platform for developing luminescent functional materials for different applications.

Preoperative ultrasound findings as risk factors of recurrence of pelvic organ prolapse after laparoscopic sacrocolpopexy.

INTRODUCTION AND HYPOTHESIS: The objective of this study is to evaluate levator ani muscle avulsion (LAMA) and ballooning as risk factors for recurrence of pelvic organ prolapse (POP) after laparoscopic sacrocolpopexy (SCP). We hypothesize that these ultrasound findings are associated with a higher risk of POP recurrence. METHODS: Retrospective cohort study of patients who underwent laparoscopic SCP between January 2015 and December 2018. Baseline translabial 3D ultrasound of the pelvic floor was performed. Pelvic Floor Distress Inventory (PFDI-20) and the Pelvic Floor Impact Questionnaire (PFIQ-7) were applied. Both univariate and multivariate analyses were carried out. RESULTS: One hundred thirty-four patients were included. On ultrasound, 32% of patients had levator ani muscle avulsion, and 36.5% had ballooning. Mean follow-up time was 16 months. There was a 13.4% anatomic recurrence; five of them (3.7%) also had symptomatic recurrence. After multivariate analysis we found that LAMA and ballooning were not significant: OR 0.99 (95% CI 0.098-10.1; p = 0.99) and OR 1.1 (95% CI 0.99-1.2; p = 0.06), respectively. CONCLUSIONS: LAMA and ballooning on pelvic floor US are not significant risk factors for anatomic POP recurrence after laparoscopic SCP. Laparoscopic SCP has a 13.4% and 3.4% anatomic and symptomatic recurrence rate, respectively, and the majority of patients reported significant improvement in quality of life.

Two-Dimensional Cartesian Coordinate System Educational Toolkit: 2D-CACSET.

Engineering education benefits from the application of modern technology, allowing students to learn essential Science, Technology, Engineering, and Mathematics (STEM) related concepts through hands-on experiences. Robotic kits have been used as an innovative tool in some educational fields, being readily accepted and adopted. However, most of the time, such kits' knowledge level requires understanding basic concepts that are not always appropriate for the student. A critical concept in engineering is the Cartesian Coordinate System (CCS), an essential tool for every engineering, from graphing functions to data analysis in robotics and control applications and beyond. This paper presents the design and implementation of a novel Two-Dimensional Cartesian Coordinate System Educational Toolkit (2D-CACSET) to teach the two-dimensional representations as the first step to construct spatial thinking. This innovative educational toolkit is based on real-time location systems using Ultra-Wide Band technology. It comprises a workbench, four Anchors pinpointing X+, X-, Y+, Y- axes, seven Tags representing points in the plane, one listener connected to a PC collecting the position of the Tags, and a Graphical User Interface displaying these positions. The Educational Mechatronics Conceptual Framework (EMCF) enables constructing knowledge in concrete, graphic, and abstract levels. Hence, the students acquire this knowledge to apply it further down their career path. For this paper, three instructional designs were designed using the 2D-CACSET and the EMCF to learn about coordinate axes, quadrants, and a point in the CCS.

Implementation of a MEIoT Weather Station with Exogenous Disturbance Input.

Due to the emergence of the coronavirus disease (COVID 19), education systems in most countries have adapted and quickly changed their teaching strategy to online teaching. This paper presents the design and implementation of a novel Internet of Things (IoT) device, called MEIoT weather station, which incorporates an exogenous disturbance input, within the National Digital Observatory of Smart Environments (OBNiSE) architecture. The exogenous disturbance input involves a wind blower based on a DC brushless motor. It can be controlled, via Node-RED platform, manually through a sliding bar, or automatically via different predefined profile functions, modifying the wind speed and the wind vane sensor variables. An application to Engineering Education is presented with a case study that includes the instructional design for the least-squares regression topic for linear, quadratic, and cubic approximations within the Educational Mechatronics Conceptual Framework (EMCF) to show the relevance of this proposal. This work's main contribution to the state-of-the-art is to turn a weather monitoring system into a hybrid hands-on learning approach thanks to the integrated exogenous disturbance input.

In vitro leishmanicidal activity and theoretical insights into biological action of ruthenium(II) organometallic complexes containing anti-inflammatories.

Leishmaniasis, a neglected tropical disease caused by protozoans of the genus Leishmania, kills around 20-30 thousand people in Africa, Asia, and Latin America annually and, despite its potential lethality, it can be treated and eventually cured. However, the current treatments are limited owing to severe side effects and resistance development by some Leishmania. These factors make it urgent to develop new leishmanicidal drugs. In the present study, three ruthenium(II) organometallic complexes containing as ligands the commercially available anti-inflammatories diclofenac (dic), ibuprofen (ibu), and naproxen (nap) were synthesized, characterized, and subjected to in vitro leishmanicidal activity. The in vitro cytotoxicity assays against Leishmania (L.) amazonensis and Leishmania (L.) infantum promastigotes have shown that complexes [RuCl(dic)(η6-p-cymene)] (1) and [RuCl(nap)(η6-p-cymene)] (3) were active against both Leishmania species. Complex [RuCl(ibu)(η6-p-cymene)] (2) has exhibited no activity. The IC50 values for the two active complexes were respectively 7.42 and 23.55 µM, for L. (L.) amazonensis, and 8.57 and 42.25 µM, for L. (L.) infantum. Based on the toxicological results and computational analysis, we proposed a correlation between the complexes and their activity. Our results suggest both complexation to ruthenium(II) and ligands structure are key elements to leishmanicidal activity.

Acetylcholinesterase Inhibitor Ameliorates Early Cardiometabolic Disorders in Fructose-Overloaded Rat Offspring.

BACKGROUND: We investigate the role of galantamine on autonomic dysfunction associated with early cardiometabolic dysfunction in the offspring of fructose-overloaded rats. METHODS: Wistar rats received fructose diluted in drinking water (10%) or water for 60 days prior to mating. Fructose overload was maintained until the end of lactation. The offspring (21 days after birth) of control and fructose-overloaded animals were divided into three groups: control (C), fructose (F) and fructose + galantamine (GAL). GAL (5 mg/kg) was administered orally until the offspring were 51 days old. Metabolic, hemodynamic and cardiovascular autonomic modulation were evaluated. RESULTS: The F group showed decreased insulin tolerance (KITT) compared to the C and GAL groups. The F group, in comparison to the C group, had increased arterial blood pressure, heart rate and sympathovagal balance (LF/HF ratio) and a low-frequency band of systolic arterial pressure (LF-SAP). The GAL group, in comparison to the F group, showed increased vagally mediated RMSSD index, a high-frequency band (HF-PI) and decreased LF/HF ratio and variance in SAP (VAR-SAP) and LF-SAP. Correlations were found between HF-PI and KITT (r = 0.60), heart rate (r = -0.65) and MAP (r = -0.71). CONCLUSIONS: GAL treatment significantly improved cardiovascular autonomic modulation, which was associated with the amelioration of cardiometabolic dysfunction in offspring of parents exposed to chronic fructose consumption.

Insulin Treatment Does Not Prevent EARLY Autonomic Cardiovascular and Diastolic Dysfunctions in Streptozotocin-Induced Diabetic Rats.

Recent studies have found increased cardiovascular mortality risk in patients with type 1 diabetes when compared to normoglycemic people, even when they were kept under good glycemic control. However, the mechanisms underlying this condition have yet to be fully understood. Using streptozotocin (STZ)-induced diabetic rats, we evaluated the effects of insulin replacement therapy on cardiac, autonomic, inflammatory, and oxidative stress parameters. Daily treatment with insulin administrated subcutaneously in the STZ-diabetic rats showed a reduction in hyperglycemia (>250 mg/dL) to normalized values. The insulin treatment was effective in preventing alterations in cardiac morphometry and systolic function but had no impact on diastolic function. Also, the treatment was not able to prevent the impairment of baroreflex-tachycardic response and systolic arterial pressure variability (SAP-V). A correlation was found between improvement of these autonomic parameters and higher levels of IL-10 and lower levels of oxidized glutathione. Our findings show that insulin treatment was not able to prevent diastolic, baroreflex, and SAP-V dysfunction, suggesting an outstanding cardiovascular risk, even after obtaining a good glycemic control in STZ-induced diabetic rats. This study shed light on a relatively large population of diabetic patients in need of other therapies to be used in combination with insulin treatment and thus more effectively manage cardiovascular risk.

A proposed model to evaluate how changes in body condition score and the fatty acid profile of a supplement affect physiology and metabolic responses of nonlactating females.

Two experiments were designed to evaluate the effects of altering body condition score (BCS) and the profile of a fatty acid (FA) supplement on the metabolism of Bos indicus Nellore females. In experiment 1, 16 and 24 B. indicus heifers and nonlactating cows, respectively, were assigned to (1) maintenance diet (MNT-MNT; n = 10), (2) maintenance diet and BCS loss (MNT-LSS; n = 10), (3) maintenance diet supplemented with calcium salts of soybean oil for 30 d and BCS loss for 40 d (MNT+CFA-LSS; n = 10), and (4) maintenance diet for 30 d and BCS loss for 40 d with a diet containing calcium salts of soybean oil (MNT-LSS+CFA; n = 10). Following the BCS loss period, MNT-LSS, MNT+CFA-LSS, and MNT-LSS+CFA were fed a diet to promote the gain of BCS. In experiment 2, 40 Bos indicus nulliparous heifers were assigned to (1) maintenance diet (MNT-MNT; n = 10), (2) BCS loss followed by a BCS gain (LSS-REM; n = 10), (3) BCS loss followed by a BCS gain diet with CFA of palm oil (LSS-REM+PLM; n = 10), and (4) BCS loss followed by a BCS gain diet with CFA of soybean oil (LSS-REM+SOY; n = 10). Blood samples were obtained for serum haptoglobin and fecal samples for pH (experiment 2 only). In experiment 1, a treatment × day interaction was observed for BCS during the 60-d BCS loss and gain period. Animals assigned to MNT-MNT had a greater BCS than the other treatment groups on d 40 and 60 of the experiment, but no other differences were observed. Moreover, a treatment × day interaction was observed for serum haptoglobin, as on d 60, MNT-LSS had a greater mean serum haptoglobin concentration. In experiment 2, a treatment × day interaction was also observed for BCS. From d -4 to 0, LSS-REM and LSS-REM+SOY had a reduced BCS versus MNT-MNT, but also lower for LSS-REM versus MNT-MNT on d 1, and LSS-REM+PLM versus MNT-MNT on d -1 and 0. For serum haptoglobin, no treatment or treatment × day interaction was observed. A treatment × day interaction was observed for fecal pH. From d -10 to 0, MNT often had a lower fecal pH, but during realimentation, LSS-REM heifers had a reduced fecal pH on d 1, 4, and 10. In summary, we failed to demonstrate an increase in serum haptoglobin due to a BCS loss. Still, supplementation with calcium salts of FA alleviated the increase in haptoglobin and maintained fecal pH at more stable values during realimentation, regardless of the FA profile of the supplement.

Effects of supplementing narasin to Bos indicus heifers during late-gestation and lactation on development of the offspring.

This experiment evaluated the effects of supplementing narasin during late-gestation and lactation on productive and physiological responses of Bos indicus beef heifers and their offspring. Pregnant, nulliparous Nelore heifers (N = 88) that conceived under the same fixed-time artificial insemination protocol and to the same sire were used. Heifers were ranked by maternal ability genomic score, body weight (BW) and body condition score (BCS) and allocated to 44 drylot pens (2 heifers per pen; 10 × 25 m). Pens were ranked by these traits and alternatively assigned to receive (NAR) or not (CON) 0.260 mg of narasin/kg of heifer BW daily (Elanco Saúde Animal, São Paulo, Brazil). Narasin was mixed into a supplement offered at 0.30% of heifer BW from day 0 until heifers weaned their calves (day 316), whereas CON heifers received the same supplement without narasin addition. Heifers received Urochloa brizantha hay and water for ad libitum consumption (days 0 to 316) and calved between days 97 to 112 of the experiment. After calving, heifers and offspring had access to hay and supplement; hence, supplements and narasin were offered according to heifer + calf BW beginning on day 162. No treatment differences were detected (P ≥ 0.18) for heifer BW and BCS during the experiment, although BW loss from day 0 to calving was less (P = 0.04) in NAR compared to CON heifers. Hay intake during the experiment did not differ (P = 0.79) between treatments. Serum IGF-I concentrations were greater (P = 0.05) for NAR heifers on day 60 of the experiment and did not differ (P ≥ 0.28) between treatments 24 h and 30 d after calving (treatment × day interaction; P = 0.04). No treatment effects were detected (P ≥ 0.58) for calf birth BW. Serum concentrations of total protein 24 h after birth were greater (P = 0.04) in calves from NAR compared with CON heifers, and a tendency (P = 0.10) for a similar outcome was noted for serum IgG concentrations. Diarrhea incidence did not differ (P = 0.16) between treatments, although the number of total diarrhea cases per calf were greater (P = 0.03) in the CON offspring. Growth rate of calves from NAR heifers tended (P = 0.08) to be greater, resulting in heavier calves at weaning (P ≤ 0.04) compared with CON offspring. Collectively, these outcomes indicate narasin supplementation to beef heifers as a nutritional alternative to improve cow-calf productivity via developmental programming effects during gestation, as well as direct consumption by their nursing offspring.

New ruthenium(II) complexes with cyclic thio- and semicarbazone: evaluation of cytotoxicity and effects on cell migration and apoptosis of lung cancer cells.

We describe the synthesis, physicochemical characterization, and in vitro antitumor assays of four novel analogous ruthenium(II) complexes with general formula cis-[RuII(N-L)(P-P)2]PF6, where P-P = bis(diphenylphosphine)methane (dppm, in complexes 1 and 2) or bis(diphenylphosphine)ethane (dppe, in complexes 3 and 4) and N-L = 5,6-diphenyl-4,5-dihydro-2H-[1,2,4]triazine-3-thione (Btsc, in complexes 1 and 3) or 5,6-diphenyltriazine-3-one (Bsc, in complexes 2 and 4). The data were consistent with cis arrangement of the biphosphine ligands. For the Btsc and Bsc ligands, the data pointed to monoanionic bidentate coordination to ruthenium(II) through N,S and N,O, respectively. Single-crystal X-ray diffraction showed that complex 1 crystallized in the monoclinic system, space group P21/c. Determination of the cytotoxicity profiles of complexes 1-4 gave SI values ranging from 1.19 to 3.50 against the human lung adenocarcinoma cell line A549 and the non-tumor lung cell line MRC-5. Although the molecular docking studies suggested that the interaction between DNA and complex 4 was energetically favorable, the experimental results showed that they interacted weakly. Overall, our results demonstrated that these novel ruthenium(II) complexes have interesting in vitro antitumor potential and this study may contribute to further studies in medicinal inorganic chemistry.

The impact of global warming on the range distribution of different climatic groups of Aspidoscelis costata costata.

The ectothermic nature of reptiles makes them especially sensitive to global warming. Although climate change and its implications are a frequent topic of detailed studies, most of these studies are carried out without making a distinction between populations. Here we present the first study of an Aspidoscelis species that evaluates the effects of global warming on its distribution using ecological niche modeling. The aims of our study were (1) to understand whether predicted warmer climatic conditions affect the geographic potential distribution of different climatic groups of Aspidoscelis costata costata and (2) to identify potential altitudinal changes of these groups under global warming. We used the maximum entropy species distribution model (MaxEnt) to project the potential distributions expected for the years 2020, 2050, and 2080 under a single simulated climatic scenario. Our analysis suggests that some climatic groups of Aspidoscelis costata costata will exhibit reductions and in others expansions in their distribution, with potential upward shifts toward higher elevation in response to climate warming. Different climatic groups were revealed in our analysis that subsequently showed heterogeneous responses to climatic change illustrating the complex nature of species geographic responses to environmental change and the importance of modeling climatic or geographic groups and/or populations instead of the entire species' range treated as a homogeneous entity.

Efficacy and safety of tension-free vaginal tape compared with transobturator tape among obese women with stress urinary incontinence: a retrospective cohort study.

OBJECTIVE: To determine the efficacy and safety of tension-free vaginal tape (TVT) compared with transobturator tape (TOT) in obese women with stress urinary incontinence (SUI). METHODS: We performed a retrospective chart review of patients who underwent insertion of TVT or TOT for stress urinary incontinence between January 2003 and October 2009. Women were excluded if they had had previous surgery for SUI or had a diagnosis of intrinsic sphincter deficiency. RESULTS: One hundred eighty obese women (BMI > 30 kg/m²) with SUI and with follow-up for at least one year were identified (90 had TVT and 90 had TOT). The rates of success on the objective criteria were 91% for the TVT group and 88% for the TOT group (P = 0.46) and 87% versus 80% (P = 0.23) on subjective assessment. CONCLUSION: Our retrospective cohort study demonstrated similar rates of cure for obese women with SUI who underwent insertion of TVT and TOT.

An in silico method to assess antibody fragment polyreactivity.

Antibodies are essential biological research tools and important therapeutic agents, but some exhibit non-specific binding to off-target proteins and other biomolecules. Such polyreactive antibodies compromise screening pipelines, lead to incorrect and irreproducible experimental results, and are generally intractable for clinical development. Here, we design a set of experiments using a diverse naïve synthetic camelid antibody fragment (nanobody) library to enable machine learning models to accurately assess polyreactivity from protein sequence (AUC > 0.8). Moreover, our models provide quantitative scoring metrics that predict the effect of amino acid substitutions on polyreactivity. We experimentally test our models' performance on three independent nanobody scaffolds, where over 90% of predicted substitutions successfully reduced polyreactivity. Importantly, the models allow us to diminish the polyreactivity of an angiotensin II type I receptor antagonist nanobody, without compromising its functional properties. We provide a companion web-server that offers a straightforward means of predicting polyreactivity and polyreactivity-reducing mutations for any given nanobody sequence.

Randomized controlled trial of cystocele plication risks: a pilot study.

OBJECTIVE: We wished to determine the feasibility of a single-centre randomized controlled trial to investigate whether avoiding cystocele plication in women undergoing transvaginal repair of cystocele decreases the need for catheterization beyond the second postoperative day. METHODS: Patients undergoing transvaginal repair of cystocele were randomly assigned to either have or not have plication sutures during their procedure. We assessed the success of recruitment, physician acceptability, and adherence to protocol, and we also assessed the need for catheterization beyond the second postoperative day. RESULTS: Twenty-two women were randomized (a recruitment rate of 85%), and participating surgeons did not feel that one surgical technique was superior to the other. There were no protocol violations after randomization. No cases of postoperative voiding dysfunction were identified. CONCLUSION: A multicentre randomized controlled trial is required to determine the short-term and long-term risks and benefits of avoiding plication sutures in women undergoing transvaginal repair of cystocele. If patients and physicians support the study protocol, conducting such a trial is feasible.

In Silico Targeting of the Long Noncoding RNA MALAT1.

RNA targeting has gained traction over the past decade. It has become clear that dysregulation of RNA can be linked to many diseases, leading to a need for new scaffolds recognizing RNA specifically. Long noncoding RNAs are emerging as key controllers of gene expression and potential therapeutic targets. However, traditional targeting methods have overwhelmingly been focused on proteins. In this study, we used a protein computational tool and found several possible targetable pockets in a structurally characterized long noncoding RNA, MALAT1. Screening against those identified pockets revealed several hit compounds. We tested the binding of those compounds to MALAT1 RNA and tRNA as a negative control, using SPR. While several compounds were nonspecific binders, others were able to recognize MALAT1 specifically. One of them, MTC07, has an apparent affinity of 400.2 ± 14.4 µM. Although it has weak affinity, MTC07 is the first compound targeting MALAT1 originating from in silico docking.

Home and Neighborhood Physical Activity Location Availability among African American Adolescent Girls Living in Low-Income, Urban Communities: Associations with Objectively Measured Physical Activity.

Based on the ecological model of active living, the neighborhood environment may relate to individual physical activity (PA) behaviors. The purposes of this study were to (1) generate a replicable neighborhood-level physical activity location availability score (PALAS) from data variables associated with physical activity among adolescents and adults, and apply this score to Baltimore City, Maryland, and (2) determine if relationships exist between PA and PA location availability. Geographic information systems (GISs) were used to create the PALAS. Using linear regression models, we examined relations between objectively measured PA among low-income, urban, predominantly African American adolescent girls (n = 555, 2009-2012 data collection), and the PALAS rating of their neighborhood environment (neighborhood PALAS) and their home neighborhood area (PALAS variables/subcomponents within 0.25 miles of the home). A PALAS map of the study area was created, illustrating neighborhoods varying in availability and variety of PA locations. After adjusting for confounders, a higher neighborhood PALAS (ß = 0.10, p = 0.041) and the presence of a recreation center in the home neighborhood area (ß = 0.46, p = 0.011) were associated with more minutes per day spent in moderate to vigorous PA. Policy makers and stakeholders should consider increasing access to PA locations as a strategy to promote PA among adolescent girls.

Evaluation of different cooling management strategies for lactating Holstein × Gir dairy cows.

Heat stress negatively impacts production, reproduction, and health of ruminants and strategies to alleviate these losses are warranted. Therefore, four experiments evaluated different cooling strategies on vaginal temperature (VT) of Holstein × Gir cows. Experiment 1 compared different amounts of water (2- or 4-L) over a 1-hour period from 1000 to 1100 h and 1600 to 1700 h. Experiment 2 evaluated the effects of sprinkling duration (in hours; 1- or 2-H), whereas Experiment 3 evaluated the effects of water amount (4- or 8-L) applied for 1- or 2-H. Lastly, the effects of a cooling strategy on specific hours of the day, starting at either 0700 (T-1) or 1100 h (T-2; Experiment 4), were evaluated. In all experiments, 12 Holstein × Gir cows were used in a 2 × 2 Latin Square Design containing two periods of 6 days each. Temperature and humidity index (THI) were recorded hourly and VT was recorded every 10-min throughout the experiments. As expected, an hour effect was observed for THI (P < 0.0001), which peaked early in the afternoon. In Experiment 1, a treatment × hour interaction was observed (P < 0.0001) for VT, as animals assigned to receive 4-L had a reduced VT at 1100, 1600, 1700, and 2300 h (P ≤ 0.03). During the cooling applications, cows receiving 4-L for 1 h had a reduced VT from 60 to 150 min (P ≤ 0.04). In Experiment 2, a treatment × hour interaction was observed (P < 0.0001) for VT, as animals assigned to receive 4-L of water for 2-H had a reduced VT at 1200 h (P = 0.05). Moreover, during the cooling process, VT was reduced for 2-H cows from 140 to 170 min after the beginning of the cooling process (P ≤ 0.05). In Experiment 3, animals assigned to receive 4-L + 2H had a reduced VT at 1200, 1700, 1800, and 1900 h (P < 0.001). A treatment × hour interaction was observed (P < 0.0001), as VT was reduced for 4-L + 2-H cows from 130 to 180 min after the beginning of the cooling process (P ≤ 0.05). In Experiment 4, by the time when the first cooling cycle of T-1 was applied (0700 h), T-1 cows consistently had (P ≤ 0.05) a reduced VT up to the hottest hours and greatest THI of the day (1400 and 1500 h). This pattern was maintained until the end of the last cooling cycle, whereas T-2 cows had a reduced VT. In summary, 4 L of water over a 5-min cycle for a period of 2 hours twice a day maintained VT of Holstein × Gir cows at lower levels. Moreover, the hour at which the first cooling cycle starts also should be considered when evaluating the efficacy of a cooling strategy for an entire day.

Impact of air pollution on pulmonary function and respiratory symptoms in children. Longitudinal repeated-measures study.

BACKGROUND: Salamanca, Mexico occupied fourth place nationally in contaminating emissions. The aim of the study was to determine the impact of air pollution on the frequency of pulmonary function alterations and respiratory symptoms in school-age children in a longitudinal repeated-measures study. METHODS: We recruited a cohort of 464 children from 6 to 14 years of age, from two schools differing in distance from the major stationary air pollution sources. Spirometry, respiratory symptoms and air pollutants (O3, SO2, NO, NO2, NOx, PM10,) were obtained for each season. Mixed models for continuous variables and multilevel logistic regression for respiratory symptoms were fitted taking into account seasonal variations in health effects according to air pollution levels. RESULTS: Abnormalities in lung function and frequency of respiratory symptoms were higher in the school closer to major stationary air pollution sources than in the distant school. However, in winter differences on health disappeared. The principal alteration in lung function was the obstructive type, which frequency was greater in those students with greater exposure (10.4% vs. 5.3%; OR = 1.95, 95% CI 1.0-3.7), followed by the mixed pattern also more frequent in the same students (4.1% vs. 0.9%; OR = 4.69, 95% CI, 1.0-21.1). PM10 levels were the most consistent factor with a negative relationship with FVC, FEV1 and PEF but with a positive relationship with FEV1/FVC coefficient according to its change per 3-month period. CONCLUSIONS: Students from the school closer to major stationary air pollution sources had in general more respiratory symptoms than those from the distant school. However, in winter air pollution was generalized in this city and differences in health disappeared. PM10 levels were the most consistent factor related to pulmonary function according, to its change per 3-month period.

Inotropic and Antiarrhythmic Transmural Actions of Ranolazine in a Cellular Model of Type 3 Long QT Syndrome. / Ações Transmurais Inotrópicas e Antiarrítmicas da Ranolazina em um Modelo Celular da Síndrome do QT Longo Tipo 3.

Ranolazine (RANO) prevents cardiac arrhythmia by blocking the late sodium current (INaL). A transmural gradient of Nav1.5 is found in the left ventricular wall of the heart. Thus, we investigated the effects of RANO in healthy cardiomyocytes and in a cellular model of type 3 long QT syndrome (LQT3). We used isolated endocardium (ENDO) and epicardium (EPI) cells and a video edge detection system and fluorescence microscopy to monitor calcium transients. RANO (0.1, 1, 10 and 30 uM, at 25oC) at a range of pacing frequencies showed a minor impact on both cell types, but RANO at 30uM and 35oC for ENDO cells attenuated sarcomere shortening by~21%. Next, to mimic LQT3, we exposed ENDO and EPI cells to anemone toxin II (ATX-II), which augments INaL. Cellular arrhythmias induced by ATX-II were abrogated by RANO (30 µM) at 35oC. Based on our results we can conclude that RANO has a minor impact on sarcomere shortening of healthy ENDO and EPI cells and it abrogates arrhythmias induced by INaLto a similar level in ENDO and EPI cells.