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Objective: To cross-validate estimates of the size of the illicit cigarette trade based on the results of four different survey methods., Methods: In 2018/2019, four non-industry-funded, large-scale studies were conducted in selected Brazilian cities: packs discarded in household garbage/PDG (1 city), packs littered in the streets/PLS (5 cities), a phone survey of tobacco users' purchase behaviors/VIGITEL (5 cities), and a face-to-face household survey of tobacco users' purchase behaviors/FTF-household (2 cities). The proportions of illicit cigarettes consumed were based on the price paid by smokers in their last purchase (VIGITEL or FTF-household) and/or direct observation of brand names and health warnings (PDG, PLS or FTF-household)., Results: Based on PLS, the share of packs that avoided taxation ranged from 30.4% (95% CI 25.6% to 35.7%) in Rio de Janeiro to 70.1% (95% CI 64.6% to 75.0%) in Campo Grande; and PDG conducted in Rio de Janeiro found an even lower proportion point estimate of illicit cigarette use (26.8%, 95% CI 25.1% to 28.6%). In FTF-household, the share of illicit cigarette consumption based on the self-reported price ranged from 29.1% (95% CI 22.4% to 35.7%) in Rio de Janeiro to 37.5% (95% CI 31.2% to 43.7%) in São Paulo, while estimates based on pack observation ranged from 29.9% (95% CI 23.3% to 36.5%) in Rio de Janeiro to 40.7% (95% CI 34.3% to 47.0%) in São Paulo. For all cities, VIGITEL presented the lowest levels of illicit consumption, and most illicit brands were produced in Paraguay., Conclusions: Small differences in the estimated levels of illicit trade across methods were found, except for the phone survey. The cross-validation of estimates from independent studies is important to help effectively implement tobacco excise tax policy in Brazil and other low-income and middle-income countries., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

The illicit trade of tobacco products, by enabling access to cheaper cigarettes, favors smoking initiation and hinders its cessation, minimizing the effects of price policy and taxes on reducing the demand for tobacco. This is especially the case among populations with lower income and schooling, where smoking is concentrated. Its confrontation requires multisectoral actions, aligned with the World Health Organization Framework Convention on Tobacco Control, supported by estimates of the illicit trade magnitude and the analysis of its characteristics. This study analyzes, based on samples of household solid waste collected in 2018 in the city of Rio de Janeiro, Brazil, the conformity of discarded cigarette packages with criteria for their regular commercialization in Brazil, classifying them as legal or illegal. The evaluation was also carried out for the 15 Planning Areas (PA) of the municipality. The Social Progress Index (SPI) was chosen to represent heterogeneities among PA. The average percentage of illegal cigarette packs found was 26.79%, ranging from 3.36% to 46.29% among PA. The PAs with high illegality presented lower Social Progress Index and lower percentages of legal cigarette packages with a price equal to or greater than BRL 7.25. Among the illegal packages, 98.07% were manufactured in Paraguay. The study contributes methodologically to measure the consumption of illegal cigarettes in the second economic capital of Brazil, supporting the National Tobacco Control Policy in the struggle against illicit trade of tobacco products and in the effective implementation of the pricing and tax policy on these products.

Acute coronary syndromes without coronary lesions have gained relevance in recent years. However, local data on this condition is scarce. We aimed to explore this entity in a National registry of acute myocardial infarction that was carried out prospectively in hospitals with cardiology residence programs from Argentina. We included 1182 patients from 45 centers, where 33 did not present coronary lesions on angiography. The mean age was 64.5 ±13.0 and 69.7% were male, without differences compared to participants with epicardial disease. The most common electrocardiographic presentation was ST segment deviation. In addition, presented lower biomarkers (peak CPK 203.5 IU / l, range [IQR] 102-422.5 vs. 895.5 IU / l IQR 350-1891, p < 0.0001). The median hospitalization was 4.0 days (IQR 3-5.5), lower than the group with intermediate and severe coronary disease (5.5 days, RIC 4-7, and 6, RIC 4-7, p = 0.003). At discharge, less use of ACE/ARB (54.6% vs.78.0% y 79.7%, p = 0.002) and statins (78.8% vs. 87.9% y 91.9%, p = 0.017). No deaths during hospitalization were reported. Our data suggested that infarcts without significant coronary lesions are frequent, although they are probably underdiagnosed. Their prognosis seems to be more favorable, but they receive fewer drugs to prevent recurrence. New studies are necessary to deepen the knowledge of the disease.

The tobacco industry normally overestimates the illegal market's size in order to reinforce the idea of its direct relationship to the choice to increase taxes by the internal revenue administration. In Brazil, the last increase in taxes on tobacco products was in 2016. The growth of demand for illegal cigarettes has other macrosocial determinants that the industry does not take into account, such as the increase in the economic capacity to purchase legal cigarettes. The article aims to test the hypothesis of the "Brazilian consumer's economic reason" from 2015 to 2019 by comparing the estimated consumptions of illegal cigarettes, based on official government data on legal production and cigarette consumption with an "extraofficial estimate" furnished by the industry. The study also used official national data on monthly income from work. The Brazilian population's "purchasing capacity for legal cigarettes" increased systematically from 2016 to 2019, from 412 packs/month to 460 packs/month. The absolute difference between the estimate by the tobacco industry and the estimate based on official data on the volume of illegal cigarettes that were consumed increased over time, reaching +30.2 billion units in 2019. Meanwhile, legal cigarette consumption, calculated with official data, increased from 2016 through 2019 (+7.8 billion), while the industry estimated a reduction in this consumption (-9.5 billion). Policymakers should seek to base their decisions on estimates generated from official data sources, including macroeconomic data on employment and income, rather than to use estimates produced by the tobacco industry that aim to interfere in public policies.

Background: A street cross-sectional survey in 2016 with a representative sample of 1697 smokers in five Colombian cities was used to estimate the penetration of illicit cigarettes (PIC). The first wave was collected 3 months before a 100% increase in tobacco excise tax, and a second wave collected data 9 months after tax reform was effective. OBJECTIVE : Analyse changes after a cigarette tax increase in PIC, prices and smoking behaviour patterns for five Colombian cities (63% of the market). Smoking behaviour includes consumption intensity, presentation (stick/pack) and place of purchase. METHODS : Repeated street cross-sectional survey with smokers' self-report on smoking behaviour, last purchase information and direct observation of smokers' packs. Sampling frame: smokers, men and women, aged 12 years or older, all income levels, resident in the five cities with the highest number of smokers representing 63% of cigarette market share (Bogotá, Medellín, Cali, Cartagena and Cúcuta) with 1 733 316 smokers in 2013. Sample size was 1697 per wave, with confidence level 95%, margin of error 3.5% for Bogotá and Medellín and 5% for the other three cities. Smokers in second wave match first wave's location, sex and age group. Illicit cigarettes were identified based on brand, health warnings and price. RESULTS : After the tax hike, the average real price of a 20-stick pack increased by 28.2% and by 23.1% for loose cigarettes. Illicit cigarettes represented 3.4% of total cigarette consumption in 2016 and increased to 6.4% in 2017, lower than the current industry estimate of 18%. Consumption intensity decreased: the proportion of heavy smokers (more than 10 cigarettes per day) wentdown from 37% in 2016 to 26% in 2017., Conclusion: After the tax increase, Colombia's PIC remained at low levels, and there is enough space for new tobacco tax hikes., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

A time-series is any set of N time-ordered observations of a process. In veterinary epidemiology, our focus is generally on disease occurrence (the "process") over time, but animal production, welfare or other traits might also be of interest. A common source of time-series datasets are animal disease monitoring and surveillance systems. Here, we scan the application of methods to analyse time-series data in the peer-reviewed, published literature. Based on this literature scan we focus on autocorrelation and illustrate the recommended steps using ARIMA (Autoregressive Integrated Moving Average Models) methods via analysis of a time-series of canine parvovirus (CPV) events in a pet dog population in Australia, 2009 to 2015. We conclude by identifying the barriers to the application of ARIMA methods in veterinary epidemiology and suggest some possible solutions. In the literature scan the selected 37 studies focused mostly on infectious and parasitic diseases, predominantly for analytical, rather than descriptive or predictive, purposes. Trends and seasonality were investigated, and autocorrelation analyzed, in most studies, most commonly using R software. An approach to analyzing autocorrelation using ARIMA methods was then illustrated using a time-series (week and month units) of CPV events in a pet dog population in Australia, reported to a national companion animal disease surveillance system. This time-series was derived by summing veterinarian reports of confirmed CPV diagnoses. We present data analysis output generated via the R statistical environment, and make this code available for the reader to apply to this or other time-series datasets. We also illustrate prediction of CPV events by rainfall as a covariate. Time-series analysis using ARIMA methods to understand and explore autocorrelation appears to be relatively uncommon in veterinary epidemiology. Some of the reasons might include limited availability of data of sufficient time unit length, lack of familiarity with analytical methods and available software, and how to best use the information generated. We recommend that wherever feasible, such time-series data be made available both for analysis and for methods development., (Copyright © 2020 Ward, Iglesias and Brookes.)

Background: Paraguay has reportedly been a major transit hub for illicit tobacco products since the 1960s, initially to supply markets in Argentina and Brazil and, more recently, other regional markets and beyond. However, to date there has been no systematic analysis, notably independent of the tobacco industry, of this trade including the roles of domestic production and transnational tobacco companies (TTCs). This article fills that gap by detailing the history of Paraguay's illicit cigarette trade to Brazil and Argentina of TTC products and Paraguayan production between 1960 and 2003. The effective control of illicit cigarette flows, under Article 15 of the World Health Organization (WHO) Framework Convention on Tobacco Control (FCTC) and the Protocol to Eliminate the Illicit Trade in Tobacco Products, requires fuller understanding of the changing nature of the illicit trade., Methods: We systematically searched internal industry documents to understand the activities and strategies of leading TTCs in Paraguay and subregion over time. We also mapped illicit trade volume and patterns using US government and UN data on the cigarette trade involving Paraguay. We then estimated Paraguay's cigarette production from 1989 to 2003 using tobacco leaf flows from the United Nations Commodity Trade Statistics Database (UN Comtrade)., Results: We identify four phases in the illicit tobacco trade involving Paraguay: 1) Paraguay as a transit hub to smuggle BAT and PMI cigarettes from the U.S. into Argentina and Brazil (from the 1960s to the mid-1970s); 2) BAT and PMI competing in north-east Argentina (1989-1994); 3) BAT and PMI competing in southern and southern-east Brazil (mid to late 1990s); and 4) the growth in the illicit trade of Paraguayan manufactured cigarettes (from the mid- 1990s onwards). These phases suggest the illicit trade was seeded by TTCs, and that the system of supply and demand on lower priced brands they developed in the 1990s created a business opportunity for manufacturing in Paraguay. Brazil's efforts to fight this trade, with a 150% tax on exports to Latin American countries in 1999, further prompted supply of the illicit trade to shift from TTCs to Paraguayan manufacturers., Conclusion: This paper extends evidence of the longstanding complicity of TTCs in the illicit trade to this region and the consequent growth of Paraguayan production in the 1990s. Our findings confirm the need to better understand the factors influencing how the illicit tobacco trade has changed over time, in specific regional contexts, and amid tobacco industry globalization. In Paraguay, the changing roles of TTC and domestic production have been central to shifting patterns of illicit supply and distribution since the 1960s. Important questions are raised, in turn, about TTCs efforts to participate as legitimate partners in global efforts to combat the problem, including a leading role in data gathering and analysis.

Background: Leading transnational tobacco companies (TTCs) began to expand their operations in Latin America in the 1960s. This included legally exporting their cigarettes to Paraguay during the 1960s which, in turn, were illegally re-exported to Argentina and Brazil. By the 1990s, competition between BAT and PMI for this lucrative illicit market, focusing on low-priced brands, prompted manufacturing in Paraguay. Paraguayan manufacturing rapidly grew after the introduction of a new cigarette export tax in Brazil in 1999., Methods: We systematically searched Truth Tobacco Industry Documents (TTID) to understand the activities and strategies of leading TTCs in Paraguay and subregion over time. We applied the analytical framework of Lee and Eckhardt (2017) to understand Tabesa's global business strategy. We searched the websites of TTCs and Tabesa for activities since the mid 2000s to understand how the companies publicly describe these strategies. We used the United Nations Commodity Trade Statistics Database (UN Comtrade) as an independent source to crosscheck statements by Tabesa executives about export markets. We contextualized and triangulated our findings with 42 key informant interviews., Results: Tabesa became the largest cigarette manufacturer in Paraguay, and one of the largest companies in the country, through complicity in the illicit trade. Enabled by market conditions created by leading TTCs, and a permissive regulatory environment in Paraguay, evidence suggests Tabesa had become a major source of illicit cigarettes across Latin America and beyond by the late 2000s. Although Brazil continues to account for the bulk of Tabesa's revenues, findings suggest that the company is aspiring to compete with TTCs in markets worldwide through legal and illegal sales., Conclusion: There is a need for fuller understanding of the risks to global tobacco control from local companies aspiring to compete with TTCs. The rise of Tabesa is part of the changing nature of the illicit trade in tobacco products which must be taken into account in implementing the Framework Convention on Tobacco Control (FCTC) and its Protocol to Eliminate Illicit Trade in Tobacco Products. Potential conflicts of interest concerning Tabesa illustrate the importance of FCTC Article 5.3 on industry interference. There is also an urgent need to address the lack of independent and rigorous data on the illicit tobacco trade in the region.

Background: By 2016, tobacco industry provided the only illicit trade estimates in Colombia and used these to discourage tax increases since the 1990s. To establish the viability of a threefold hike in the excise tax, policy makers needed unbiased estimates of the illicit cigarette., Objective: To estimate the size of illicit cigarette trade in five Colombian cities (63% of the market), analyse characteristics of smokers of illicit cigarettes and compare market share results with one industry-funded survey., Methods: Street cross-sectional survey with smokers' self-report on consumption pattern, last purchase information and direct observation of smoker's packs. Sampling frame: smokers, men and women, 12 years old or older, all income levels, resident in five Colombian cities (Bogotá, Medellín, Cali, Cartagena and Cúcuta) with 1 733 316 smokers in 2013. Sample size 1697, simple random sample by city, sampling weights based on age groups and cities. Confidence level 95%, margin of error 3.5% for Bogotá and Medellín and 5% for the other three cities. Data collection period: 24 August-14 September 2016., Results: Illicit cigarettes represent 3.5% of consumption in the five cities, a much lower estimate than the industry data. There are significant differences across cities, with Bogotá at the bottom (1.5%) and Cúcuta at the top (22.8%)., Conclusion: The low overall penetration of illicit cigarettes in Colombia indicates that the industry's warnings against tax increases are not justified. The limited importance of tax levels as determinant of consumption of illicit cigarettes is also suggested by the differences across cities, all of them with the same tax regime., Competing Interests: Competing interests: None declared., (© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

Objectives: To estimate the proportions of illicit cigarette consumption in Brazil from 2012 to 2016, a period of sharp increases in cigarette taxes., Methods: We used an approach based on legal sales provided by the Secretariat of Federal Revenues and self-reported consumption data from an annually conducted telephone survey (VIGITEL) to estimate the changes over time in illegal cigarette use in Brazil. For that purpose, we also used available information on the proportion of illegal cigarette consumption from a nationwide household survey conducted in 2013 to calculate a constant proportion of underreporting from VIGITEL in relation to total consumption and sales in Brazil., Results: There was an increase in the estimated proportion of illicit cigarette use from 2012 to 2013 (from 28.6% to 32.3%), then a decrease from 2013 to 2014 (32.3% to 28.8%), and then a sustained trend of increase from 2014 to 2016 (28.8% to 42.8%)., Conclusions: Novel and feasible approaches to estimate changes over time in the illegal market are important for helping the effective implementation of tobacco excise tax policy.

Background: Brazil experienced a large decline in smoking prevalence between 2008 and 2013. Tax rate increases since 2007 and a new tobacco tax structure in 2012 may have played an important role in this decline. However, continuous tax rate increases pushed up cigarette prices over personal income growth and, therefore, some consumers, especially lower income individuals, may have migrated to cheaper illicit cigarettes., Objective: To use tobacco surveillance data to estimate the size of illicit tobacco consumption before and after excise tax increases., Methods: We defined a threshold price and compared it with purchasing prices obtained from two representative surveys conducted in 2008 and 2013 to estimate the proportion of illicit cigarette use among daily smokers. Generalised linear model was specified to understand whether the absolute difference in proportions over time differed by sociodemographic groups and consumption levels. Our findings were validated using an alternative method., Results: Total proportion of illicit daily consumption increased from 16.6% to 31.1% between 2008 and 2013. We observed a pattern of unadjusted absolute decreases in cigarette smoking prevalence and increases in the proportion of illicit consumption, irrespective of gender, age, educational level, area of residence and amount of cigarettes consumed., Conclusions: The strategy of raising taxes has increased government revenues, reduced smoking prevalence and resulted in an increased illicit trade. Surveillance data can be used to provide information on illicit tobacco trade to help in the implementation of WHO Framework Convention on Tobacco Control (FCTC) article 15 and the FCTC Protocol to Eliminate Illicit Trade in Tobacco Products., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.)

Brazil has experienced a large decline in cigarette consumption in the last 25years. However, the most recent annual reports from the tobacco industry market leader in Brazil did not show a decrease in its gross profits. This is particularly important because tobacco industry donations/sponsorships come directly from the industry's reported gross-profits and are used to subvert health policies. The aim of the present study was to estimate (i) tobacco industry's gross-profit from legal cigarettes sales, and (ii) all-cause smoking-attributable deaths (SADs) among current Brazilian smokers who consumed legal cigarettes in 2013. We collected information on prevalence of legal cigarette use, cigarette consumption, price per cigarette pack among individuals aged ≥35years from the Global Adult Tobacco Survey, legal cigarettes sales (e.g., average costs and total volume of sales) provided by the Federal Secretariat of Revenues, and population mortality from the available vital statistics. With a gross-profit of US$1.378 billion (1.307-1.434) from sales of 54.6 billion sticks of cigarettes (53.4-55.5) to 8,424,510 smokers aged 35years and older in Brazil in 2013, cumulative SADs were estimated at 96,012 (85,647-107,654) (around 34% of cumulative SADs also including current smoking of illegal cigarettes and past smoking), i.e., one SAD was equivalent to a gross-profit of US$14,352 (12,140-16,743). Our results revealed the association between sales of cigarettes, gross-profits, and deaths in Brazil. As tobacco industry donations/sponsorships originate from industry's gross profits, which, in turn, depend on cigarette sales, our findings may be useful for increasing "moral pressure" on individuals and institutions and help countries in stopping tobacco industry interference in health policies., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The Brazilian cigarette excise tax reform of 2011 increased tax rates significantly in the presence of a high proportion of illegal and cheap cigarettes contributing to total consumption. Prior to 2011, tobacco tax policy in Brazil had reduced excise tax share on consumer prices, for fear of smuggling. This report examines two hypotheses explaining why tax authorities changed direction. The first is related to lack of concern regarding smuggling in tobacco industry pricing behavior before 2011 (rather than reducing prices following tax reduction, legal companies increased net of tax prices above inflation and key costs). The second hypothesis regards inconsistent industry assessments of the size of the illicit market, which ultimately undermined the credibility of the industry with tax authorities. The author concludes that the 2011 reform was designed to revert the weakness of previous policies, and did indeed succeed. The post-2011 experience in Brazil indicates that increased cigarette excise taxes can increase government revenues and reduce smoking prevalence and consumption despite widespread smuggling of tobacco products.

The time delay to detection of an outbreak of an emergency animal disease directly affects the size of the outbreak at detection and the likelihood that the disease can be eradicated. This time delay is a direct function of the efficacy of the surveillance system in the country involved. Australia has recently completed a comprehensive review of its general surveillance system examining regional variation in both the behaviour of modelled outbreaks of foot and mouth disease and the likelihood that each outbreak will be detected and reported to government veterinary services. The size of the outbreak and the time delay from introduction to the point where 95% confidence of detection was reached showed significant (p < 0.05) regional variation with the more remote northern areas experiencing smaller outbreaks that are less likely to spread and less likely to be reported to government services than outbreaks in the more developed southern areas of Australia. Outbreaks in the more densely populated areas may take up to 43 days until a 95% confidence of detection is achieved and at that time, the outbreak may involve up to 53 farms., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Australia, as a relatively isolated country with a high level of agricultural production, depends on, and has the opportunity to maintain, freedom from a range of important diseases of livestock. Occasional incursions of such diseases are generally detected by 'passive', general surveillance (GS). In current surveillance planning, a risk-based approach has been taken to optimising allocation of resources to surveillance needs, and having mapped the relative risk of introduction and establishment of diseases of concern, a means of mapping the efficacy of GS for their detection was required, as was a means of assessing the likely efficacy of options for improving GS efficacy if needed. This paper presents the structure and application of a tool for estimating the efficacy of Australia's GS, using the example of foot and mouth disease (FMD). The GS assessment tool (GSAT) is a stochastic spreadsheet model of the detection, diagnosis and reporting of disease on a single infected farm. It utilises the output of an intraherd disease spread model to determine the duration and prevalence of infection on different types of farm. It was applied separately to each of twelve regions of Australia, demarcated by dominant livestock production practices. Each region supplied estimates of probabilities relevant to the detection of FMD, for each of fourteen farm types and all species susceptible to the disease. Outputs of the GSAT were the average probability that FMD on the farm would be detected (single farm sensitivity), the average time elapsed from incursion of the disease to the chief veterinary officer (CVO) being notified (time to detection), and the number of average properties that would need to be infected before the CVO could be 95% confident of detecting at least one. The median single farm sensitivity for FMD varied among regions from 0.23 to 0.52, the median time to detection from 20 to 33 days, and the number of properties infected for 95% confidence of detecting at least one from 4 to 12. The GSAT has proved a valuable tool in planning surveillance for detection of exotic livestock disease in Australia, and it provides a practical example of the use of probabilistic modelling to answer important questions in the face of imperfect information., (Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.)

When induced to ovulate during anoestrus, ewes, does and cows frequently develop a short-lived corpora lutea (SLCL) syndrome associated to lack of previous progesterone. Exogenous progesterone precludes SLCL by blocking oxytocin endometrial receptors, thus inducing normal life-span CL (NLCL). Paradoxically, circa 50% of unprimed ewes do not develop SLCL. We report results from 3 trials assessing follicular, oestrous, ovulatory, and luteal end-points after 17β-oestradiol or MAP treatments. Oestradiol benzoate (50μg) induced follicular turnover, provoked ovulation in 40% (24/60) of ewes treated (93% of which developed SLCL), but did not affect the incidence of SLCL (26/53) after an allogenic sexual stimulation (ASS) by rams and oestrous ewes. By the onset of the ASS, most NLCL ewes (26/27) had already experienced turnover of their largest follicle, had smaller largest and second largest follicles, and ovulated their largest follicle more frequently than SLCL ewes did. Most SLCL ewes (19/25) did not ovulate their largest follicles, ovulating instead smaller follicles of identical size to those of NLCL ewes. Priming (40mg of MAP for 12 days) was partially effective at preventing SLCL even when terminated 14 days in advance of an ASS, but failed at completely preventing SLCL when terminated 6 or more days in advance. The coupling of a timed acquisition of full steroidogenic capability before ovulation with a system of endometrial oestradiol-progesterone-oxytocin receptors linked in an unstable equilibrium controlling the amplification of the luteolytic feed-forward loop of oxytocin and prostaglandin F(2)α explains occurrence and relative incidences of both NLCL and SLCL, and links proximate and ultimate causes of the SLCL syndrome., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Pannexin1 (Panx1), a protein related to the gap junction proteins of invertebrates, forms nonjunctional channels that open upon depolarization and in response to mechanical stretch and purinergic receptor stimulation. Importantly, ATP can be released through Panx1 channels, providing a possible role for these channels in non-vesicular signal transmission. In this study we expressed exogenous human and mouse Panx1 in the gap junction deficient Neuro2A neuroblastoma cell line and explored the contribution of Panx1 channels to cell-cell communication as sites of ATP release. Electrophysiological (patch clamp) recordings from Panx1 transfected Neuro2A cells revealed membrane conductance that increased beyond 0 mV when applying voltage ramps from -60 to +100 mV; threshold was correlated with extracellular K+, so that at 10 mM K+, channels began to open at -30 mV. Evaluation of cell-cell communication using dual whole cell recordings from cell pairs revealed that activation of Panx1 current in one cell of the pair induced an inward current in the second cell after a latency of 10-20 s. This paracrine response was amplified by an ATPase inhibitor (ARL67156, 100 μM) and was blocked by the ATP-degrading enzyme apyrase (6.7 U/ml), by the P2 receptor antagonist suramin (50 μM) and by the Panx1 channel blocker carbenoxolone. These results provide additional evidence that ATP release through Panx1 channels can mediate nonsynaptic bidirectional intercellular communication. Furthermore, current potentiation by elevated K+ provides a mechanism for enhancement of ATP release under pathological conditions.

Domestic ruminant selectivity induces floristic changes in pasturelands, risking sustainability and limiting the subsequent availability of susceptible plant species. Development of preferences for species of lower nutritional quality may help to overcome those problems. In this study, we tested the hypothesis that early experience of sheep with a low-quality food (LQF) in a nutritional enriched context increases preference for LQF in adulthood. We predicted a higher proportional consumption of LQF in experienced lambs (EL) than in inexperienced lambs (IL) in choice situations involving LQF and alternative foods. Additionally, we determined intake of LQF by EL and IL at different levels of high-quality food (HQF) availability. From 60 to 210 days of age, EL were fed in separated feed bunks mature oat hay (LQF) simultaneously with sunflower meal (SM) and corn grain (CG), whereas IL were fed alfalfa hay (HQF) simultaneously with SM and CG. After exposure, EL and IL were offered LQF in free choice situations involving alternative foods, and also at five levels of HQF availability (100%, 75%, 50%, 25% and 0% of ad libitum intake). Proportional consumption of LQF was lower or similar in EL than IL. Intake of LQF was also lower or similar in EL than IL at all levels of HQF availability, except when the LQF was the only food available. Our results did not support the hypothesis that early experience with a LQF in a nutritional enriched context increases preference for LQF in adulthood. On the contrary, experience with LQF diminished subsequent preference for LQF in adulthood. It is proposed that, in the conditions of our study, continuous comparison between the LQF and the high-quality supplements (CG and SM) during the early exposure period lead to devaluation of LQF by EL through a simultaneous negative contrast effect.

Genotype effects on lamb growth, survival, and commercial finishing were estimated in a 5-yr study aimed at assessing potential benefits from introducing meat breeds into extensive sheep systems of northeastern Patagonia. Five ram [Corriedale: CO; Border Leicester: BL; Ile de France: IF; Texel: TX; and synthetic CRIII (25% Merino, 37.5% IF, 37.5% TX)] and 5 dam (CO; synthetic CRIII; BLCO: BL x CO; IFCO: IF x CO; and TXCO: TX x CO) genotypes were represented in the study. Data were collected from 1,426 born and 1,258 weaned lambs of 9 resulting genotypes (CO x CO, BL x CO, IF x CO, TX x CO, CRIII x CO, CRIII x BLCO, CRIII x IFCO, CRIII x TXCO, and CRIII x CRIII). Birth weight was recorded on all lambs; subsequently, BW and BCS (scale 1 to 5) were recorded at regular intervals until weaning. Body weights were adjusted to 60 and 90 (weaning) d of age, and ADG were calculated from the adjusted BW for the periods 0 to 60 d and 60 to 90 d; BCS was adjusted to 90 d. Survival to weaning and percentage of lambs reaching commercial finishing (BW >or= 23 kg and BCS >or= 2.5 points) were also recorded. Significant (P < 0.05) genotype x litter size interactions were detected for birth weight and ADG 60 to 90 d. With the exception of CRIII x CO, crossbred and synthetic genotypes presented greater (P < 0.05) ADG 0 to 60 d and BW at 60 and 90 d than CO x CO lambs. Second cross lambs reared as singles presented greater ADG 60 to 90 d (P < 0.05) than BL x CO, TX x CO, and CRIII x CO, but less (P < 0.05) ADG 60 to 90 d, and no differences were observed for twins. The IF x CO, CRIII x BLCO, CRIII x IFCO, and CRIII x CRIII genotypes showed greater BCS at 90 d (P < 0.05) than CO x CO. The probability of greater commercial finishing for crossbred and synthetic genotypes relative to CO x CO was at least 79%. Probabilities of greater survival to weaning in CO x CO and CRIII x BLCO lambs relative to IF x CO, TX x CO, CRIII x CO, and CRIII x CRIII lambs were greater than 81%. Results indicate significant improvements in lamb BW and saleable lambs from the introduction of meat genotypes. Among the terminal sire breeds evaluated, BL and IF would produce the greatest impact. Differential nutritional management of pregnant ewes carrying twins and of twin lambs beyond 60 d should be implemented to mitigate litter size x genotype interactions constraining growth potential benefits, which may be critical for northeastern Patagonia conditions.

Two experiments were conducted in consecutive years in which recently (Experiment 1) or temporarily (Experiment 2) weaned ewes and matched post-partum non-lactating flockmates (DRY) were exposed to a stimulus group of rams and oestrous ewes (10 and 20 in Experiment 1, 20 and 20 in Experiment 2) for 28 days in spring. Lactating ewes (n = 130) in Experiment 1 were isolated from their lambs 4 (W-4), 2 (W-2), 1 (W-1) or 0 (W-0) days in advance and exposed along with a group of 32 DRY flockmates. Lactating ewes in Experiment 2 (n = 230) were allocated to an unreplicated factorial of two levels of temporary weaning before stimulation (B0: control; B24: lambs removed 24 h before stimulation) by four levels of ewe-lamb contact imposed at the start of the stimulation (A0: control; A12, A24 and A36: lamb-ewe separation during the initial 12, 24 or 36 h of exposure); DRY ewes (n = 54) acted as an augmented factorial control. Oestrus (rump marks) and ovulation (laparoscopy on day 5 and on day 28 (Experiment 1) or day 32 (Experiment 2)) were recorded. Ovulation and oestrous responses in Experiment 1 were similar for DRY (90.6% and 55.2%, respectively) and recently weaned ewes (83.8% and 53.7%, respectively). Amongst recently weaned ewes, the immediate ovulation response to the rams and the proportion of ewes still cycling by day 28 tended to be lower (P = 0.065 and P = 0.011) in ewes weaned on the day of ram exposure (71.9% and 54.8% v. 87.8% and 80.0%, respectively). Ovulation rate was lower (P < 0.003) in W-2 ewes (1.3 ± 0.10) than in the other recently weaned groups. In Experiment 2, ovulation (83.3%) and oestrous (68.9%) responses in DRY ewes were higher (P = 0.022 and P = 0.053, respectively) than in lactating ewes (66.2% and 51.0%, respectively). More ewes ovulated (P = 0.036) in B24 (70.5%) than in B0 (61.8%). Ewes having their lambs returned 12 h after the onset of stimulation (A12) had poorer ovulation responses (54.9%) than control ewes (A0, 72.9%, P < 0.05); this was probably associated to lamb restitution after the sunset. Main conclusions were that (i) the presence of the lambs is a depressing factor of both ovulation and oestrous responses to the ram effect in lactating ewes, (ii) the ovulation response of lactating ewes will probably benefit from removing lambs for a period of 24 h before the onset of stimulation, (iii) until additional information becomes available, temporary weaning protocols should be designed avoiding lamb restitution during the night.

Serial blood sampling before and after exposing four anovular Corriedale ewes to a group of rams and estrous ewes during the non-breeding season revealed a pattern of LH secretion similar to that previously observed in Merinos. Mean LH values doubled (P<0.001) from 0.24+/-0.06 microgL(-1) (mean+/-s.e.m.) before to 0.55+/-0.05 microgL(-1) after 2h of visual, auditory, and odor exposure to rams and estrous ewes in an indoor facility. A non-significant (P<0.17) increase of LH pulses per hour was also observed (0.7+/-0.3 pulses per hour before compared with 1.3+/-0.3 during stimulation). All four ewes had recently formed corpora lutea by five days after stimulation. Results are consistent with the pattern of sudden increase and sustained release of LH observed in other sheep breeds, particularly the Merino.

Zoning is a key prescriptive tool for administration and management of protected areas. However, the lack of zoning is common for most protected areas in developing countries and, as a consequence, many protected areas are not effective in achieving the goals for which they were created. In this work, we introduce a quantitative method to expeditiously zone protected areas and we evaluate its ecological implications on hypothetical zoning cases. A real-world application is reported for the Talampaya National Park, a UNESCO World Heritage Site located in Argentina. Our method is a modification of the zoning forest model developed by Bos [Bos, J., 1993. Zoning in forest management: a quadratic assignment problem solved by simulated annealing. Journal of Environmental Management 37, 127-145.]. Main innovations involve a quadratic function of distance between land units, non-reciprocal weights for adjacent land uses (mathematically represented by a non-symmetric matrix), and the possibility of imposing a connectivity constraint. Due to its intrinsic spatial dimension, the zoning problem belongs to the NP-hard class, i.e. a solution can only be obtained in non-polynomial time [Nemhausser, G., Wolsey, L., 1988. Integer and Combinatorial Optimization. John Wiley, New York.]. For that purpose, we applied a simulated annealing heuristic implemented as a FORTRAN language routine. Our innovations were effective in achieving zoning designs more compatible with biological diversity protection. The quadratic distance term facilitated the delineation of core zones for elements of significance; the connectivity constraint minimized fragmentation; non-reciprocal land use weightings contributed to better representing management decisions, and influenced mainly the edge and shape of zones. This quantitative method can assist the zoning process within protected areas by offering many zonation scheme alternatives with minimum cost, time and effort. This ability provides a new tool to improve zoning within protected areas in developing countries.