Rotational and ro-vibrational spectroscopy of small nitrogen-containing species and their astrophysical applications

This thesis presents the study of small nitrogen-bearing molecules, from diatomic radicals to complex organic molecules, by means of rotational and ro-vibrational spectroscopy. Besides their theoretical relevance, which spans from anharmonic force field analyses to energetic and structural properties, I have chosen this family of species because of their astrochemical importance. After some basic knowledge of molecular spectroscopy and astrochemistry is introduced, the instrumentation used during the course of my PhD school is described. Then, the most relevant studies I conducted during the last three years are presented. Generally speaking, a number of molecules of astrophysical relevance have been characterized by means of rotational and ro-vibrational spectroscopy. The sample of studied species is constituted by small radicals (imidogen, amidogen, and titanium nitride), cyanopolyynes (cyanoacetylene) and pre-biotic molecules (aminoacetonitrile): these studies are presented in great detail. Among the results, the first astronomical detection of two deuterated radicals (NHD and ND2) is presented in this thesis.Thanks to our studies, it was possible to clearly identify molecular absorptions of these species towards the pre-stellar core IRAS16293-2422, as recorded by the Herschel Space Observatory mission. These observations confirm the strong deuterium enhancement generally observed in this cloud but they reveal that models underestimate the abundances of NHD and ND2. I also report the detection of vibrationally excited aminoacetonitrile (NH2CH2CN) in Sagittarius B2, as observed in the ReMoCa survey. This is the second detection of aminoacetonitrile in the interstellar medium and the first astronomical observation of its vibrationally hot lines. This represents a small step toward the comprehension on how complex organic molecules are formed and which processes can lead to the formation of glycine. Finally, few general remarks are discussed and the importance of future laboratory studies is pointed out, along with possible perspectives.

Preclinical evaluation of a MYCN-specific inhibitor for the treatment of small cell lung cancer

Small cell lung cancer (SCLC) is the most aggressive form of lung cancer, characterized by rapid growth, early metastasis and acquired drug resistance. SCLC is usually sensitive to initial treatment, however, most patients relapse within few months; thus more effective therapies are urgently needed. Key genetic alterations very frequently observed in SCLC include loss of TP53 and RB1 and mutations in the MYC family genes (MYC, MYCL or MYCN). One of them is amplified and overexpressed in a mutually exclusive manner and represents the most prominent activating oncogene alteration in this malignancy. In particular, MYCN amplification is associated with tumor progression, treatment failure and poor prognosis. Given the role of MYCN in SCLC and its restricted expression profile, MYCN represents a promising therapeutic target; although it is considered undruggable by traditional approaches. An innovative approach to target the oncogene concerns specific MYCN expression inhibition, acting directly at the level of DNA, through an antigene peptide nucleic acid (agPNA) oligonucleotide, called BGA002. This thesis focused on the study of BGA002, as a possible targeted therapeutic strategy for the treatment of MYCN-related SCLC. In this context, BGA002 proved to be a specific and highly effective inhibitor. Furthermore, MYCN silencing induced alterations in many downstream pathways and led to apoptosis, in concomitance with autophagy reactivation. Moreover, systemic administration of BGA002 was effective in vivo as well, significantly increasing survival in MNA mouse models, even in the scenario of multidrug-resistance. In addition, BGA002 treatment successfully reduced N-Myc protein expression and, more importantly, caused a massive diminishment in tumor vascularization in the multidrug-resistant model. Overall, these results proved that MYCN inhibition by BGA002 may represent a new promising precision medicine approach, to treat MYCN-related SCLC.

Acoustic backscatter of small pelagic fish species in the Adriatic Sea

The aim of the present study is to apply a broad range of techniques to increase the knowledge of acoustic properties of Sprattus sprattus, Scomber colias and Trachurus mediterraneus in the Adriatic Sea. A novel study using tethered live fish but not involving hooks and anesthetic was tested on T. mediterraneus and S. colias through several ex situ experiments using a split-beam scientific echosounder operating at 38, 120, and 200 kHz. The mean TS was estimated for 29 live specimens, resulting in a conversion factor b20 value of -71.4 dB re 1 m2 and -71.6 dB re 1 m2 respectively which is ~3 dB lower than the current one in use in the Mediterranean Sea. Successively, two monospecific trawl hauls were analyzed through the application of in situ approach for the computation of TS values of S. sprattus which led to six b20 values for sprat (range, -68.8 dB re 1 m2 to -65.6 dB re 1 m2), all higher than the current known value of -71.7 dB re 1 m2. The high difference up to 4.2 dB compared to the current value translates in a significant decrease of absolute sprat biomass along the time series un to 20%. Finally, 149 specimens of the three species were collected for backscattering model application(i.e. Kirchhoff-ray mode model (KRM) and Finite Element Method (FEM)) from digital images of the fish body and swimbladder obtained from Computer Tomography (CT) and X-Ray scans. The values resulting from the application of KRM and FEM are in agreement with empirical results. In general terms the present work proposes the acoustic backscatter characterization of S. colias, S. sprattus and T. mediterraneus in the Mediterranean Sea.

Comparative genetic study confirms exceptionally low genetic variation in the ancient and endangered relictual conifer, Wollemia nobilis (Araucariaceae)

The Wollemi pine, Wollemia nobilis (Araucariaceae), was discovered in 1994 as the only extant member of the genus, previously known only from the fossil record. With fewer than 100 trees known from an inaccessible canyon in southeastern Australia, it is one of the most endangered tree species in the world. We conducted a comparative population genetic survey at allozyme, amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) loci in W. nobilis, Araucaria cunninghatnii and Agathis robusta - representatives of the two sister genera. No polymorphism was detected at 13 allozyme loci, more than 800 AFLP loci or the 20 SSR loci screened in W. nobilis. In Ag. robusta only one of 12 allozyme loci, five of 800 AFLP loci and none of the 15 SSR loci were variable. For A. cunninghamii, 10 of > 800 AFLP loci and five of 20 SSR loci were variable. Thus low genetic diversity characterizes all three species. While not ruling out the existence of genetic variation, we conclude that genetic diversity is exceptionally low in the Wollemi pine. To our knowledge this is the most extreme case known in plants. We conclude that the combination of small population effects, clonality and below-average genetic variation in the family are probable contributing factors to the low diversity. The exceptionally low genetic diversity of the Wollemi pine, combined with its known susceptibility to exotic fungal pathogens, reinforces current management policies of strict control of access to the pines and secrecy of the pine locations.

Extensive variation at MHC DRB in the New Zealand sea lion (Phocarctos hookeri) provides evidence for balancing selection.

Marine mammals are often reported to possess reduced variation of major histocompatibility complex (MHC) genes compared with their terrestrial counterparts. We evaluated diversity at two MHC class II B genes, DQB and DRB, in the New Zealand sea lion (Phocarctos hookeri, NZSL) a species that has suffered high mortality owing to bacterial epizootics, using Sanger sequencing and haplotype reconstruction, together with next-generation sequencing. Despite this species' prolonged history of small population size and highly restricted distribution, we demonstrate extensive diversity at MHC DRB with 26 alleles, whereas MHC DQB is dimorphic. We identify four DRB codons, predicted to be involved in antigen binding, that are evolving under adaptive evolution. Our data suggest diversity at DRB may be maintained by balancing selection, consistent with the role of this locus as an antigen-binding region and the species' recent history of mass mortality during a series of bacterial epizootics. Phylogenetic analyses of DQB and DRB sequences from pinnipeds and other carnivores revealed significant allelic diversity, but little phylogenetic depth or structure among pinniped alleles; thus, we could neither confirm nor refute the possibility of trans-species polymorphism in this group. The phylogenetic pattern observed however, suggests some significant evolutionary constraint on these loci in the recent past, with the pattern consistent with that expected following an epizootic event. These data may help further elucidate some of the genetic factors underlying the unusually high susceptibility to bacterial infection of the threatened NZSL, and help us to better understand the extent and pattern of MHC diversity in pinnipeds.

Contemporary pollen flow, mating patterns and effective population size inferred from paternity analysis in a small fragmented population of the Neotropical tree Copaifera langsdorffii Desf. (Leguminosae-Caesalpinioideae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Discovery of the first wild population of the small red brocket deer Mazama bororo (Artiodactyla: Cervidae).

Mazama bororo was described from a few captive specimens in Brazil by cytogenetic and morphological characters. These specimens supposedly originated in the Southern Atlantic Forest; however, no wild population has been reported. This study was initiated in 1998 to investigate the presence of this species in forest remnants of the Paranapiacaba mountain range, south São Paulo State, Brazil. Five specimens were captured between 2000 and 2002. Cytogenetic analysis from blood samples confirmed its specific identification, documenting the first population of small red brocket deer at the Intervales State Park.

Consumption Diagnosis of Sweet Cassava in three Municipalities in Cuba

Cassava is an important staple food for human and animal feeding in Cuba. Despite its importance, there is little or nonexistent information to diagnose preferences and frequency of consumption of cassava in that country. In this sense, the present article characterizes the preferences and frequency of consumption of cassava in the municipalities of Plaza de la Revolución-La Habana province, El Salvador–Guantanamo province and San José de Las Lajas–Mayabeque province in Cuba. A survey was conducted through a questionnaire containing twelve closed and two open questions. The sample was determined based on the number of total population of each municipality considering 95% as confidence interval and 5% as error margin. The results were statistically analyzed by calculating the absolute and the relative frequencies of each question. It was observed that the acquisition of cassava in the municipalities of Plaza de la Revolución, El Salvador and San José de las Lajas in Cuba is done by purchase small quantities of fresh cassava for home consumption within one week, due to the extreme perishability of cassava, which limits consumers' ability to store fresh roots at home. The choice of cassava is made based on both skin colour (light brown) and pulp (white) and empirical knowledge about its ease of cooking, and that cassava is mostly consumed in boiled and fried forms up to four times a week in times where there is root market supply with the desirable culinary characteristics (cooking facility), that is, from September to December.

Small-scale genetic structure of Cerastoderma glaucum in a lagoonal environment: potential significance of habitat discontinuity and unstable population dynamics

Environmental heterogeneity in coastal lagoons is expected to facilitate local adaptation in response to different ecological conditions, causing significant genetic structuring within lagoon populations at a small scale and also differentiation between lagoons. However, these patterns and processes of genetic structuring are still poorly understood. The aims of our study were (1) to seek genetic structure at a small scale in Cerastoderma glaucum inside the Mar Menor coastal lagoon using a mitochondrial DNA marker (COI) that has previously detected genetic differentiation inside the lagoon in other species and (2) to evaluate the influence of extreme environmental conditions and habitat discontinuity on its genetic composition. The results indicate high levels of haplotype diversity and low values of nucleotide diversity. COI data provide evidence of significant population differentiation among some localities within the lagoon. Limited gene flow and unstable population dynamics (i.e. fluctuations in population size caused by local extinction and recolonization), probably due to the high environmental heterogeneity, could generate the small-scale genetic divergence detected between populations within the lagoon.

Population structure of the lancelet Branchiostoma caribaeum (Cephalochordata: Branchiostomidae) in the Baía de Guanabara, Rio de Janeiro, southeastern Brazil

Population structure of the lancelet Branchiostoma caribaeum Sandevall, 1853 was studied in four surveys, corresponding to austral seasons, in a tropical bay, southeast of Brazil. Abundance was higher in the spring and was positively correlated to coarse sediments, limiting its occurrence to some sectors of the sampling area. Body length and biomass differed seasonally but not between sexes. Sexually mature individuals occurred in all seasons, suggesting continuous breeding that is typical of tropical species. Variation in the frequency of small specimens indicates temporal differences in the intensity of breeding. The body length of recruits differed from other population of lancelets and the small length which B. caribaeum attained sexual maturity in Guanabara Bay may be related to local environmental stress or the great availability of food.

Successful maintenance of a stingless bee population despite a severe genetic bottleneck

Stingless bees play an important ecological role as pollinators of many wild plant species in the tropics and have significant potential for the pollination of agricultural crops. Nevertheless, conservation efforts as well as commercial breeding programmes require better guidelines on the amount of genetic variation that is needed to maintain viable populations. In this context, we carried out a long-term genetic study on the stingless bee Melipona scutellaris to evaluate the population viability consequences of prolonged breeding from a small number of founder colonies. In particular, it was artificially imposed a genetic bottleneck by setting up a population starting from only two founder colonies, and continued breeding from it for a period of over 10 years in a location outside its natural area of occurrence. We show that despite a great reduction in the number of alleles present at both neutral microsatellite loci and the sex-determining locus relative to its natural source population, and an increased frequency in the production of sterile diploid males, the genetically impoverished population could be successfully bred and maintained for at least 10 years. This shows that in stingless bees, breeding from a small stock of colonies may have less severe consequences than previously suspected. In addition, we provide a simulation model to determine the number of colonies that are needed to maintain a certain number of sex alleles in a population, thereby providing useful guidelines for stingless bee breeding and conservation efforts.

Small area population disease burden

Small area health statistics has assumed increasing importance as the focus of population and public health moves to a more individualised approach of smaller area populations. Small populations and low event occurrence produce difficulties in interpretation and require appropriate statistical methods, including for age adjustment. There are also statistical questions related to multiple comparisons. Privacy and confidentiality issues include the possibility of revealing information on individuals or health care providers by fine cross-tabulations. Interpretation of small area population differences in health status requires consideration of migrant and Indigenous composition, socio-economic status and rural-urban geography before assessment of the effects of physical environmental exposure and services and interventions. Burden of disease studies produce a single measure for morbidity and mortality - disability adjusted life year (DALY) - which is the sum of the years of life lost (YLL) from premature mortality and the years lived with disability (YLD) for particular diseases (or all conditions). Calculation of YLD requires estimates of disease incidence (and complications) and duration, and weighting by severity. These procedures often mean problematic assumptions, as does future discounting and age weighting of both YLL and YLD. Evaluation of the Victorian small area population disease burden study presents important cross-disciplinary challenges as it relies heavily on synthetic approaches of demography and economics rather than on the empirical methods of epidemiology. Both empirical and synthetic methods are used to compute small area mortality and morbidity, disease burden, and then attribution to risk factors. Readers need to examine the methodology and assumptions carefully before accepting the results.

Sexual maturity and egg production in an unexploited population of the red squat lobster Pleuroncodes monodon (Decapoda, Galatheidae) from Central America

The squat lobster Pleuroncodes monodon is one of the most common and abundant galatheid species in the deepwater fishery of Pacific coast of America. In contrast with Peru and Chile, the red squat lobster is not commercially exploited in Costa Rica. Here we estimated the size of sexual maturity and analysed egg production in P. monodon from the Pacific of Costa Rica; the results might be important for an adequate management strategy when initiating a sustainable fishery of this resource in Costa Rica. Analyses of allometric growth revealed that females and males reached their morphological sexual maturity at 26.9 and 30.4 mm CL. respectively. The percentage of immature females captured in the present study (54%) suggests that almost half of females are able to reproduce before they are caught. Egg production was positively correlated with body size; females that measured between 24.1 and 29.0 mm CL contributed roughly 90% to the offspring production of the population. Females from Costa Rica had a considerably lower average egg volume (0.039 mm(3)) and higher fecundity (9810) than females from Chile. In comparison to other American galatheids P. monodon from Costa Rica produces numerous but small eggs, a typical strategy of species with a prolonged larval development. An analysis between our results and information published about P. monodon from Chile also revealed differences in the size-frequency distribution shape of both populations. The non-exploited character of the Costa Rican P. monodon population and a latitudinal (temperature-related) cline in body size might be associated with these differences. (C) 2010 Elsevier B.V. All rights reserved.

Lack of access and continuity of adult health care: a national population-based survey

OBJECTIVE To describe the lack of access and continuity of health care in adults.METHODS A cross-sectional population-based study was performed on a sample of 12,402 adults aged 20 to 59 years in urban areas of 100 municipalities of 23 states in the five Brazilian geopolitical regions. Barriers to the access and continuity of health care and were investigated based on receiving, needing and seeking health care (hospitalization and accident/emergency care in the last 12 months; care provided by a doctor, by other health professional or home care in the last three months). Based on the results obtained by the description of the sample, a projection is provided for adults living in Brazilian urban areas.RESULTS The highest prevalence of lack of access to health services and to provision of care by health professionals was for hospitalization (3.0%), whilst the lowest prevalence was for care provided by a doctor (1.1%). The lack of access to care provided by other health professionals was 2.0%; to accident and emergency services, 2.1%; and to home care, 2.9%. As for prevalences, the greatest absolute lack of access occurred in emergency care (more than 360,000 adults). The main reasons were structural and organizational problems, such as unavailability of hospital beds, of health professionals, of appointments for the type of care needed and charges made for care.CONCLUSIONS The universal right to health care in Brazil has not yet been achieved. These projections can help health care management in scaling the efforts needed to overcome this problem, such as expanding the infrastructure of health services and the workforce.

Injuries and envenoming by aquatic animals in fishermen of Coxim and Corumbá municipalities, state of Mato Grosso do Sul, Brazil: identification of the causative agents, clinical aspects and first aid measures

INTRODUCTION: The fishes of continental Brazil have socioeconomic importance due to their potential for sport fishing and commercial and subsistence uses, as seen in the Upper Paraguay River Basin, particularly in the municipalities of the Pantanal region, where it is the second largest economic activity. Injuries caused in professional fishermen are common and poorly studied, as in other regions of the country. METHODS: Data were obtained from questionnaires and interviews with 100 professional fishermen, 50 in each municipality, between December 2008 and October 2009. RESULTS: All the fishermen reported some kind of injury caused by fish stings (78% of injuries) and fish, alligator and snake bites (22%) on the hands (46% of cases) and feet (35% of cases). Most of the patients had mild symptoms. The most severe cases were associated with secondary bacterial infections and required specific treatment and prolonged recovery associated with social and economic losses. CONCLUSIONS: The results of this study indicate that the stressful work conditions, inattention to basic preventive measures and carelessness were factors that contributed to accidents and that the toxicity and ability to inflict mechanical trauma of some aquatic species, plus the ineffective use of first aid and hospital treatment, contributed to the high morbidity and complications in many cases. Data from this study are relevant to the fishing communities of the Pantanal region, since they reveal high rates of accidents, lack of knowledge concerning first aid, initial treatment, injury prevention and lack of medical follow-up of the population.