Resumo A hibernação é uma condição natural dos animais que vivem na zona temperada, embora alguns lagartos tropicais também experenciem hibernação anualmente, como é o caso do lagarto nativo da América do Sul, Salvator merianae ou “teiú”. Embora as características fisiológicas e metabólicas associadas à hibernação tenham sido amplamente estudadas, possíveis alterações na integridade das hemácias durante esse período ainda permanecem obscuras. A desidratação e o jejum são consequências naturais da hibernação por vários meses e podem estar relacionadas a algumas modificações celulares. Neste estudo, investigamos se a tolerância osmótica de hemácias do lagarto teiú sob hibernação são diferentes das células obtidas de animais em atividade normal. Além disso, investigamos indiretamente por meio da quantificação de biomoléculas oxidadas e da atividade de enzimas antioxidantes se a membrana das hemácias dos teiús em hibernação poderia estar associada à oxidação. Nossos resultados sugerem que as hemácias possuem maior fragilidade durante o período de hibernação, embora não tenhamos encontrado evidências de um cenário de estresse oxidativo associado à essa fragilidade acentuada. Embora não tenhamos excluído a possibilidade de dano oxidativo durante a hibernação, sugerimos que um aumento no volume das hemácias como consequência de sangue hipoosmótico durante a hibernação também poderia afetar a integridade de hemácias, tal como foi observado.

Abstract Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or “tegu” lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

ABSTRACT Arenas Blancas is a poorly known fossiliferous site located in the lower reach of the Chasicó creek (Buenos Aires Province, Argentina), with great relevance from a biostratigraphic viewpoint. The Macrochorobates scalabrinii Biozone was defined in this site, proposed as the biostratigraphic basis of the early Huayquerian Stage/Age (Late Miocene); however, the geological context and faunal record of this site have never been studied in detail. In this work, we perform a multi-proxy analysis of the Arenas Blancas site, as well as a nearby site here called Curva de la Vaca, and provide new interpretations on their origin, age, and biostratigraphy. Sedimentological, stratigraphic, and geomorphological characteristics of both sites suggest that the sequences include fluvial/alluvial deposits that represent Quaternary terraces. The taxonomic analysis of the Arenas Blancas faunal assemblage evidences the presence of 14 mammal taxa, together with some fishes, reptiles, and birds; the assemblage is correlated with the assemblage from the Cerro Azul Formation assigned to the Chasicoan Stage/Age (Late Miocene), also recovered from the lower reach of the Chasicó creek. The use of the taxa proposed in previous works to characterize the Macrochorobates scalabrinii Biozone (including this species) is not supported due to their taxonomic status and/or temporal distribution. Taphonomic evaluation indicates that the assemblage is constituted by reworked specimens; in this frame, it is proposed that fluvio/alluvial events, occurred during the Late Pleistocene-Holocene, eroded the Upper Miocene substrate and reworked fossil remains. Based on the present evidence, the Macrochorobates scalabrinii Biozone is here rejected as a valid biostratigraphic unit.

RESUMEN Arenas Blancas es una localidad fosilífera poco conocida, ubicada en el curso inferior del arroyo Chasicó (provincia de Buenos Aires, Argentina), pero con gran relevancia desde el punto de vista bioestratigráfico. La Biozona Macrochorobates scalabrinii , propuesta como base bioestratigráfica del Piso/Edad Huayqueriense temprano (Mioceno tardío) fue definida en este sitio, aunque su contexto geológico y registro faunístico nunca han sido estudiados en detalle. En este trabajo analizamos el sitio Arenas Blancas desde diferentes perspectivas, como así también un sitio cercano aquí denominado Curva de la Vaca, a fin de aportar nuevas interpretaciones sobre su origen, edad y bioestratigrafía. Las características sedimentológicas, estratigráficas y geomorfológicas de ambos sitios sugieren que las secuencias aflorantes corresponden a depósitos fluviales/aluviales que representan terrazas cuaternarias. El análisis taxonómico de la asociación faunística de Arenas Blancas permitió reconocer 14 taxones de mamíferos, como así también peces, reptiles y aves. Esta asociación muestra una clara correlación con la asociación de la Formación Cerro Azul asignada al Piso/Edad Chasiquense (Mioceno tardío), también registrada en el curso inferior del arroyo Chasicó. Por otra parte, se considera que el uso de los taxones que caracterizan la Biozona Macrochorobates scalabrinii (incluyendo esta especie) no está avalado por su estatus taxonómico y/o su distribución temporal. El estudio tafonómico muestra que la asociación está constituida por especímenes retrabajados; en este contexto, se propone que los eventos fluviales/aluviales ocurridos durante el Pleistoceno tardío-Holoceno erosionaron el sustrato del Mioceno tardío y retrabajaron los restos fósiles. Sobre la base de la evidencia disponible, se rechaza la validez de la Biozona Macrochorobates scalabrinii como unidad bioestratigráfica.

Abstract Introduction Studies have shown that the loss of the FOXO3 transcriptional function is involved in the pathophysiology of some chronic erythroid disorders, including beta-thalassemia (β-thal). Therefore, the single nucleotide polymorphism (SNP) rs3800231 (35-2764A > G) could contribute to alterations in its transcriptional activity, acting as a modifier of β-thal phenotypic manifestations. Objective and method In order to better understand the genotypic and/or allelic distributions among β-thal patients, we evaluated 83 β-thal heterozygous and 20 homozygous, compared to 117 individuals without hemoglobinopathies (control group). Additionally, we verified any influence of the FOXO3 polymorphism on clinical manifestations among β-thal homozygotes. Results We obtained higher frequencies of the wild-type homozygous (AA) and the wild-type allele (A) in the β-thal group (p< 0.0001 and p= 0.00014, respectively). The most common clinical manifestations found among β-thal homozygotes were iron overload (90%), splenomegaly (65%) and bone complications (35%), e.g., osteopenia/osteoporosis. We observed that close to 80% of the patients presenting such manifestations had the genotype AA. However, we did not find any significant involvement of the FOXO3 polymorphism in clinical manifestation occurrences. Conclusion Thus, we concluded that the SNP rs3800231 did not play a significant role as a modifier of the clinical manifestations observed in the β-thal homozygotes studied.

Abstract Two allopatric populations of Brazilian diploid and tetraploid Odontophrynus americanus species complex, both from São Paulo state, had their blood hemoglobin biochemically analyzed. In addition, these specimens were cytogenetically characterized. Biochemical characterization of hemoglobin expression showed a distinct banding pattern between the allopatric specimens. Besides this, two distinct phenotypes, not linked to ploidy, sex, or age, were observed in adult animals of both populations. Phenotype A exhibits dark-colored body with small papillae, ogival-shaped jaw with reduced interpupillary distance and shorter hind limbs. Phenotype B shows yellowish-colored body with larger papillae, arch-shaped jaw with broader interpupillary distance and longer hind limbs. Intermediate phenotypes were also found. Considering the geographical isolation of both populations, differences in chromosomal secondary constrictions and distinct hemoglobins banding patterns, these data indicate that 2n and 4n populations represent cryptic species in the O. americanus species complex. The observed phenotypic diversity can be interpreted as population genetic variability. Eventually future data may indicate a probable beginning of speciation in these Brazilian frogs. Such inter- and intrapopulational differentiation/speciation process indicates that O. americanus species complex taxonomy deserves further evaluation by genomics and metabarcoding communities, also considering the pattern of hemoglobin expression, in South American frogs.

RESUMO O sistema de integração lavoura-pecuária (SILP) é uma alternativa que vem atraindo grande interesse dos agricultores nas regiões produtoras de grãos, devido ao suporte à pecuária por meio da produção de alimento, além de melhoria nas propriedades físicas do solo. O objetivo deste trabalho foi avaliar o efeito do pastejo e da adubação nitrogenada em atributos físicos do solo e na produtividade da cultura do milho, em um sistema de integração lavoura-pecuária. O delineamento foi em blocos ao acaso, com parcelas subdivididas, sendo as parcelas as doses de nitrogênio, e as subparcelas, a presença e ausência de pastejo. Foram utilizadas três repetições por tratamento. Os tratamentos foram doses de nitrogênio de 0; 75; 150 e 225 kg ha-1 aplicados na pastagem de azevém (Lolium multiflorum Lam.) e aveia-branca (Avena sativa Lam.) no inverno. Posteriormente à pastagem, foi implantada a cultura do milho (Zea mays Lam.) no verão. O pastejo com controle da taxa de lotação animal de ovinos e manutenção da massa de forragem consorciada com aveia e azevém não alterou a densidade, a macroporosidade, a microporosidade e a porosidade total no Latossolo Bruno. O pastejo nas áreas que receberam adubação nitrogenada na pastagem, não prejudicou a produtividade da cultura do milho. A adubação nitrogenada da pastagem de inverno, igual ou superior a 150 kg N ha-1, garante elevada produtividade da cultura do milho na ausência da aplicação de N.

ABSTRACT Grain farmers have seen integrated crop-livestock systems (ICLS) as an alternative of great interest by providing cattle food supply, as well as improvement of the phisical properties of soil. This study aimed at evaluating the effect of grazing and nitrogen levels on soil physical properties and on corn yield in an integrated crop-livestock farming. The experimental design was a randomized complete block design with a split plot arrangement of treatments and three replications per treatment; nitrogen doses were the main plots, and grazing presence or absence were the subplots. We applied four doses of nitrogen (0, 75, 150, and 225 kg ha-1) in soils grown with ryegrass (Lolium multiflorum Lam.) and oat (Avena sativa Lam.) during winter. After the grazing period, the area was planted with corn (Zea mays Lam.) within summer. Sheep grazing with stocking rate control and herbage mass maintenance using oats and ryegrasses did not change soil density, macro, micro and total porosity in a Bruno Latosol (Oxisol). Grazing in nitrogen-fertilized areas did not impair corn yield. Previous nitrogen fertilization in winter grazing, in amounts equal to or higher than 150 kg N ha-1, ensures high corn yields in the absence of N applications.

O entendimento da interação entre os fatores que compõem o sistema de integração lavoura-pecuária pode direcionar o uso sustentável do solo e das pastagens. Foi estudada a influência do pastejo e doses de nitrogênio em atributos físicos do solo (densidade, macroporosidade, microporosidade, porosidade total e resistência à penetração) e na produtividade da cultura do feijoeiro, em um sistema de integração lavoura-pecuária. O delineamento foi em blocos ao acaso, com parcelas subdivididas e 3 repetições. Os tratamentos foram as doses de nitrogênio de 0; 75; 150 e 225 kg ha-1, na pastagem de azevém (Lolium multiflorum Lam.) no inverno; e na sequência o cultivo do feijoeiro (Phaseolus Vulgaris Lam.), no verão. As subparcelas caracterizaram presença ou ausência de pastejo. No pastejo com ovinos, em sistema de lotação contínua, a manutenção da massa de forragem de azevém e a adubação nitrogenada não comprometem a qualidade física do solo. A adubação nitrogenada, antecipada na pastagem de inverno (azevém) garante elevada produtividade do feijoeiro como cultura sucessora, sem necessidade de reaplicação de N, sendo recomendada a dose de 150 kg ha-1 de N na pastagem. O efeito do pisoteio do ovino na resistência do solo à penetração ficou restrito a 0,10 m superficiais do solo. O Pastejo durante o inverno permite alta produtividade do feijoeiro mesmo em baixas doses de nitrogênio.

Knowing the interaction among factors that compound integrated crop-livestock system can provide a sustainable use of soil and pasture. It was evaluated grazing and nitrogen doses influence on soil physical properties including density, macroporosity, microporosity, total porosity, mechanical penetration resistance, and beans yield of an integrated crop-livestock system. The experiment was performed in a split-plot randomized block design, with three replications. Treatments were four nitrogen levels: 0, 75, 150 and 225 kg ha-1 on ryegrass (Lolium multiflorum Lam) in winter with common bean (Phaseolus vulgaris Lam.) in summer. Additionally, subplots were grazing presence or absence. Sheep continuous grazing under adequate ryegrass forage maintenance and nitrogen fertilization did not compromise soil physical quality. Nitrogen fertilization at 150 kg ha-1 rate during winter pasture (ryegrass) allows subsequent high bean yield, without the need of another nitrogen fertilization. Sheep treading effect on soil penetration resistance was greater at 0.10 m of depth. The winter grazing during allows high bean yield even at low nitrogen doses.

As hemoglobinopatias e talassemias constituem as afecções genéticas mais comuns, apresentando-se, na maioria dos casos, em heterozigose. Diante da diversidade de hemoglobinas variantes encontrada na população brasileira, metodologias específicas e complementares para um diagnóstico laboratorial preciso, capaz de elucidar possíveis interações entre estas variantes genéticas, são necessárias. Este relato de caso descreve a interação entre hemoglobina B2 e a hemoglobina S em um indivíduo do sexo feminino, caucasoide, proveniente da região Sudeste do Brasil, identificada por meio de técnicas eletroforéticas em diferentes pH, cromatografia líquida de alta performance e PCR- RFLP. Visto que a hemoglobina B2 coelui com a hemoglobina S na análise cromatográfica e dificilmente é visualizada em eletroforese pH alcalino, devido à sua baixa concentração, justifica-se a necessidade da associação de testes laboratoriais, inclusive moleculares, na rotina do diagnóstico de hemoglobinas para a correta identificação do perfil de hemoglobinas do indivíduo e real frequência na população brasileira. Rev. Bras. Hematol. Hemoter.

Hemoglobinopathies and thalassemias are the most common genetic diseases, and in most cases, present as heterozygous. Due to the diversity of hemoglobin variants, specific and complementary methodologies are necessary for a precise laboratorial diagnosis, able to elucidate possible interactions between genetic polymorphisms. This case report describes an interaction between hemoglobin B2 and hemoglobin S in a Caucasian woman from the southeastern region of Brazil. This interaction was identified by electrophoresis in different pHs, high performance liquid chromatography and PCR-RFLP. As hemoglobin B2 is eluted in the same window as hemoglobin S in automatic HPLC systems and is hardly seen in alkaline electrophoresis due to its low concentration, the association must be confirmed using additional laboratorial tests, including molecular biology techniques. These tests should be included in the routine practice of hemoglobinopathy diagnosis in order to correctly identify hemoglobin variants and to know the real frequency of these mutations in the Brazilian population. Rev. Bras. Hematol. Hemoter.

Variations in the phenotypic expression of heterozygous beta thalassemia reflect the formation of different populations. To better understand the profile of heterozygous beta-thalassemia of the Brazilian population, we aimed at establishing parameters to direct the diagnosis of carriers and calculate the frequency from information stored in an electronic database. Using a Data Mining tool, we evaluated information on 10,960 blood samples deposited in a relational database. Over the years, improved diagnostic technology has facilitated the elucidation of suspected beta thalassemia heterozygote cases with an average frequency of 3.5% of referred cases. We also found that the Brazilian beta thalassemia trait has classic increases of Hb A2 and Hb F (60%), mainly caused by mutations in beta zero thalassemia, especially in the southeast of the country.

A low iron level, the commonest nutritional deficiency in the world, is a public health problem in developing countries. On the other hand, an excessive amount of iron is toxic, causing several organic dysfunctions, such as diabetes, cirrhosis, endocrinopathies and heart disease. Researchers have reported an association of iron overload with beta-thalassemia. The aim of this paper was to compare the serum ferritin levels of women with the beta-thalassemia trait. The results of serologic tests of 137 women of childbearing age were analyzed; 63 had the beta-thalassemia trait and 74 had Hb AA. In the beta-thalassemia carriers, the median ferritin value was 51.90 ng/mL and in the non-carriers 31.60 ng/mL (p = 0.0052). Levels of less than 20 and above 150 ng/mL were observed in 28% and 3% of the non-carriers and in 16% and 11% of the carriers, respectively. With these results it is possible to conclude that women in the reproductive age with the beta-thalassemia trait present higher ferritin levels in the northeastern region of São Paulo State. Further studies are necessary to clarify possible genetic and/or environment factors which interfere in iron absorption.

As hemoglobinopatias são um grupo de afecções genéticas que representam problema de saúde pública em muitos países em que sua incidência é alta, com significativa morbidade. Objetivamos identificar defeitos moleculares que pudessem explicar o perfil laboratorial obtido por eletroforese e HPLC com Hb F elevada, em um grupo de indivíduos adultos sem sinais ou sintomas de anemia. Encontramos cinco diferentes mutações que originam beta talassemia por PCR-ASO: três casos com CD 6 (-A), um CD 39, um IVS 1-5, um -87 todas de origem mediterrânea, e um IVS II-654 de origem asiática. As mutações CD 6 (-A), -87 e IVS II-654 foram descritas pela primeira vez na população brasileira.

Hemoglobinopathies are a heterogeneous group of genetic disorders which represent a public health problem, with significant morbidity, in countries where the prevalence is high. This study aimed at identifying molecular abnormalities that might explain the laboratorial profile obtained using electrophoresis and high performance liquid chromatography in a group of individuals without signs or clinical symptoms of anemia. Five different mutations for beta-thalassemia were found using PCR-ASO: three cases with CD 6 (-A), one CD 39, one IVI I-6, one -87 (mutations originating in the Mediterranean region) and one IVS II-654 (mutation originating in Asia). This is the first time that the CD 6 (-A), -87 and IVS II-654 mutations have been described in the Brazilian population.

RESUMO A utilização de plantas inseticidas tem se mostrado uma ferramenta promissora para controle de insetos, contudo ainda há muito pouco no que diz respeito ao efeito sobre Alphitobius diaperinus (Panzer). Portanto, o objetivo do trabalho foi avaliar a atividade inseticida de extratos vegetais visando o controle desse inseto em aviários. Foram testados extratos etanólicos (E), diclorometânicos (D), hexânicos (H) e aquosos (A) de Annonamuricata, Chenopodiumambrosioides, Eucalyptus grandis, Melia azedarach, Ocimum basilicum, Ruta graveolens e Tagetes erecta, e um óleo comercial à base de nim, Azadirachta indica (Dalneem®). Os extratos e o óleo foram diluídos na concentração de 10% e pulverizados sobre os insetos, mantidos em placas de Petri. O controle recebeu aplicação de água destilada. Os tratamentos mais eficientes foram óleo comercial de A. indica (97,5%), extrato de R. graveolens (E) (61,3%), C. ambrosioides (H) (32,5%) e de folhas de M. azedarach (E e D) (ambos 26,3%). Comparou-se também a aplicação direta do óleo sobre o inseto e aplicação na superfície com a qual o inseto ficaria em contato, sendo maior a eficiência quando pulverizado sobre os insetos (100%) em relação ao contato com superfície tratada (40,9%). Comparando-se diferentes concentrações do óleo pulverizado nos insetos, observou-se relação direta entre atividade inseticida e concentração utilizada, não havendo efeito apenas na menor concentração (0,1%). Contudo, o óleo não apresentou atividade sobre os insetos quando aplicado na cama do aviário.

ABSTRACT The use of insecticidal plants seems to be a promising method for insect control, however there are few studies in relation to their effect on Alphitobius diaperinus (Panzer). The present study was carried out to evaluate the insecticidal activity of plant extracts for the alternative control of this insect in poultry houses. Tests were made using ethanolic (E), dichlorometanic (D), hexanic (H) and aqueous (A) extracts from Annona muricata, Chenopodium ambrosioides, Eucalyptus grandis, Melia azedarach, Ocimum basilicum, Ruta graveolens andTagetes erecta, and a commercial oil from Azadirachta indica (Dalneem®). Extracts and oil were diluted at 10% and sprayed on the insects, kept in Petri dishes, and the control consisted of distilled water. The most effective treatments were: A. indica commercial oil (97.5%), extracts from R. graveolens (E) (61.3%), C. ambrosioides (H) (32.5%) and leaf extracts from M. azedarach (E and D) (both 26.3%). The direct application of the neem oil on the insect was also tested in comparison to its application on the surface with which the insect would be in contact;in this case the efficiency was higher when sprayed on the insects (100%) in relation to the contact with treated surface (40.9%). Comparing different concentrations of the neem oil sprayed on the insects, a direct relation between insecticidal activity and concentration was observed, although no effect was observed at the lowest concentration (0.1%). However, the neem oil did not present activity on the insects when applied in the poultry litter.

The most highly prevalent inherited disease in Brazil and in the world, sickle cell anemia, is considered a public health problem. Characterized by homozygosis for the hemoglobin S gene, the individual has a range of signs and symptoms that require careful treatment. The sickle cell trait is characterized by heterozygosis for the hemoglobin S gene, however the carrier does not express the disease. In the current study we aimed at verifying the presence of the sickle cell trait in 1000 blood donors of the Hematology and Hemotherapy Center of the State of Piauí (Hemopi) in the period from October 2007 to April 2008. After analysis by alkaline and acid electrophoresis, positive cases were confirmed by molecular biology. We obtained rates of 3.4% for hemoglobin AS and 5% for hemoglobin AC, with a total frequency of 3.9% in the total of 1,000 blood donors.