The venom proteome of the poorly studied desert coral snake Micrurus tschudii tschudii was unveiled using a venomic approach, which identified ≥38 proteins belonging to only four snake venom protein families. The three-finger toxins (3FTxs) constitute, both in number of isoforms (~30) and total abundance (93.6% of the venom proteome), the major protein family of the desert coral snake venom. Phospholipases A2 (PLA2s; seven isoforms, 4.1% of the venom proteome), 1–3 Kunitz-type proteins (1.6%), and 1–2 l-amino acid oxidases (LAO, 0.7%) complete the toxin arsenal of M. t. tschudii. Our results add to the growing evidence that the occurrence of two divergent venom phenotypes, i.e., 3FTx- and PLA2-predominant venom proteomes, may constitute a general trend across the cladogenesis of Micrurus. The occurrence of a similar pattern of venom phenotypic variability among true sea snake (Hydrophiinae) venoms suggests that the 3FTx/PLA2 dichotomy may be widely distributed among Elapidae venoms.

We report a structural and functional proteomics characterization of venoms of the two subspecies (Bothrops bilineatus bilineatus and B. b. smaragdinus) of the South American palm pit viper from the Brazilian state of Rondônia and B. b. smaragdinus from Perú. These poorly known arboreal and mostly nocturnal generalist predators are widely distributed in lowland rainforests throughout the entire Amazon region, where they represent an important cause of snakebites. The three B. bilineatus spp. venom samples exhibit overall conserved proteomic profiles comprising components belonging to 11 venom protein classes, with PIII (34–40% of the total venom proteins) and PI (8–18%) SVMPs and their endogenous tripeptide inhibitors (SVMPi, 8–10%); bradykinin-potentiating-like peptides (BBPs, 10.7–15%); snake venom serine proteinases (SVSP, 5.5–14%); C-type lectin-like proteins (CTL, 3–10%); phospholipases A2 (PLA2, 2.8–7.6%); cysteine-rich secretory proteins (CRISP, 0.9–2.8%); l-amino acid oxidases (LAO, 0.9–5%) representing the major components of their common venom proteomes. Comparative analysis of the venom proteomes of the two geographic variants of B. b. smaragdinus with that of B. b. bilineatus revealed that the two Brazilian taxa share identical molecules between themselves but not with Peruvian B. b. smaragdinus, suggesting hybridization between the geographically close, possibly sympatric, Porto Velho (RO, BR) B. b. smaragdinus and B. b. bilineatus parental populations. However, limited sampling does not allow determining the frequency of this event. The toxin arsenal of the South American palm pit vipers may account for the in vitro recorded collagenolytic, caseinolytic, PLA2, l-amino acid oxidase, thrombin-like and factor X-activating activities, and the clinical features of South American palm pit viper envenomings, i.e., local and progressively ascending pain, shock and loss of consciousness, spontaneous bleeding, and profound coagulopathy. The remarkable cross-reactivity of the Brazilian pentabothropic SAB antivenom toward the heterologous B. b. bilineatus venom suggests that the paraspecific antigenic determinants should have been already present in the venom of the last common ancestor of the Bothrops ″jararaca″ and ″taeniatus″ clades, about 8.5 Mya in the mid-late Miocene epoch of the Cenozoic era. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifiers PXD020043, PXD020026, and PXD020013.

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Abstract

Introduction

In 2004, Peru ratified the Health Organization (WHO) Framework Convention on Tobacco Control (FCTC) and in 2006 passed Law 28705 for tobacco consumption and exposure reduction. The Global Youth Tobacco Survey (GYTS) provides data on youth tobacco use for development of tobacco control programs. Findings from the GYTS conducted in four main cities in Peru in 2000 and 2003 are reported in this paper and can be used to monitor provisions of the WHO FCTC.

Methods

The GYTS is a school-based survey that uses a standardized methodology for sampling, questionnaire construction, field procedures, and data management. In total, 5,332 and 7,824 students aged 13 to 15 years participated in the 2000 and 2003 surveys conducted in Huancayo, Lima, Tarapoto and Trujillo.

Results

In both years, Lima had the highest lifetime (54.6% and 59.6%) and current use of tobacco (18.6% and 19.2%) of the four cities. According to gender, boys smoked more than girls and less than 20% of students initiated smoking before the age of 10. Among smokers, more than 60% bought their cigarettes in a store with no restriction for their age, and approximately 12% had ever been offered "free cigarettes". Around 90% of students were in favor of banning smoking in public places. Changes between 2000 and 2003 included an increase in the percentage of smokers who wanted to have a cigarette first thing in the morning in Tarapoto (from 0% to 1.2%) and a decrease in exposure to tobacco at home in Huancayo (from 23.7% to 17.8%) and Trujillo (from 27.8% to 19.8%)

Conclusion

While few changes in tobacco use among youth have been observed in the GYTS in Peru, the data in this report can be used as baseline measures for future evaluation efforts. At this time, tobacco control efforts in Peru need to focus on enhancing Law 28705 to include enforcement of existing provisions and inclusion of new laws and regulations. Most of these provisions are required of all countries, such as Peru, that have ratified the WHO FCTC.

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