Interventions to prevent urinary catheter-associated infections in children and neonates: a systematic review.

INTRODUCTION: Few data are available to inform strategies for the prevention of catheter-associated urinary tract infection (CAUTI) in children and neonates. Many recommendations are derived from studies in adults and cannot be applied to the paediatric population. OBJECTIVE: This study was aimed to identify all studies that measured the efficacy of an intervention for the prevention of CAUTI in children and neonates. METHODS: A systematic review using the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-analyses) was conducted. Eligible studies published between January 1st, 1995 and December 31st, 2017, were identified in PubMed, the Cochrane Database of Systematic Reviews, LILACS, SciELO and DOAJ if applying an intervention with the aim of CAUTI prevention in inpatient children, infants or neonates. The following study designs were included: controlled and non-controlled before-and-after studies, (controlled) interrupted time series analyses and randomized controlled trials. Quantitative or qualitative studies on interventions in both adults and children were eligible if data on children could be extracted. Reviews, case series, letters, notes, conference abstracts and opinion articles were excluded. RESULTS: Of 99 articles identified, six were included in the final analysis, after consensus from three independent investigators. Four studies used a multimodal strategy (using at least four or more different components at the same time) as follows: aseptic rules during catheter insertion and removal; cleaning the urethral meatus with sterile water; use of a new silicone catheter per insertion with a closed sterile drainage system by a sterile technique; daily evaluation of catheter requirement; placement of indwelling urinary catheters only for approved indications; reducing of urinary catheter days and positioning of the patient and collection device to assist in urine drainage. One study tested periurethral cleaning intervention to reduce CAUTI. One study described the association of the presence of a physician safety champion with urinary catheter device utilization ratios. Catheter-associated UTI reduction rates were reported in four studies; three achieved statistically significant decreases in CAUTI rates. Positive results were achieved only when a multimodal strategy was used with at least four or more components. This strategy could be adopted for paediatric healthcare institutions to reduce CAUTI rates in children and neonates. CONCLUSION: Evidence exists to support the use of a multimodal strategy for CAUTI reduction in hospitalized children and neonates.

Role of antimicrobial stewardship programmes in children: a systematic review.

The United Nations and the World Health Organization have designated antimicrobial resistance (AMR) as a major health priority and developed action plans to reduce AMR in all healthcare settings. Establishment of institutional antimicrobial stewardship programmes (ASPs) is advocated as a key intervention to reduce antibiotic consumption in hospitals and address high rates of multi-drug-resistant (MDR) bacteria. PUBMED and the Cochrane Database of Systematic Reviews (January 2007-March 2017) were searched to identify studies reporting the effectiveness of ASPs in general paediatric wards and paediatric intensive care units (PICUs) for reducing antibiotic consumption, use of broad-spectrum/restricted antibiotics, and antibiotic resistance and healthcare-associated infections (HAIs). Neonatal units and antifungal agents were excluded. Of 2509 titles and abstracts, nine articles were eligible for inclusion in the final analysis. All studies reported a reduction in the use of broad-spectrum/restricted antibiotics or antibiotic consumption. One study reported a reduction in HAIs in a PICU, and another study evaluated bacterial resistance, showing no effect following ASP implementation. Prospective audit on antibiotic use was the most common ASP core component (eight of nine studies). Antibiotic pre-authorization was described in two studies. Other described interventions were the provision of guidelines or written information (five of nine studies), and training of healthcare professionals (one study). There is limited evidence for a reduction in antibiotic consumption and use of broad-spectrum/restricted agents following ASP implementation specifically in PICUs. Data evaluating the impact of ASPs on HAIs and AMR in PICUs are lacking. In addition, there is limited information on effective components of a successful ASP in PICUs.

Genetic diversity of arboreal cotton populations of the Brazilian semiarid: a remnant primary gene pool for cotton cultivars.

Mocó cotton belong to the same species as the cultivated species, Gossypium hirsutum, and cultivated forms were mainly landraces but also developed as cultivars, bearing good fiber quality and drought tolerant when cropped as a perennial species. The northeast Brazil crop system based on this cotton type is finished, with a few small area planted in the three main States, where it was previously cultivated (Ceará, Paraíba, and Rio Grande do Norte), but in others, maintenance is accomplished by single dooryard plants. Plants were found in all visited Northeast Brazil municipalities, sometimes in the North of the country, and were collected for ex situ preservation and evaluation. Most of seeds had no fuzz (62.2%) and 94.6% of the genotypes presented spot in flowers. Seventy-one alleles were revealed in 12 loci. The genetic structure of the population evaluated by microsatellite markers shows two main groups, one comprising the Seridó region where landraces were originated and other comprising the state of Ceará, where a specific breeding program was developed. Genotypes collected in North Brazil States as well as those collected in Bahia, Alagoas, and Sergipe grouped with those collected in Ceará. The Mantel correlogram indicates a significant (P < 0.05) correlation between genetic and geographical distances up to 77 km. The ex situ maintenance and agronomical evaluation are the main concerns for mocó, as the use of the agricultural interesting traits, possibly introgressed to other genotypes, is predicted. The in situ preservation is still of interest since there is more diversity there than in the collected plants and some should be continued due to use as medicinal plant.

Using atmospheric plasma to increase wettability, imbibition and germination of physically dormant seeds of Mimosa Caesalpiniafolia.

In this study, we analyzed seed wettability as well as imbibition and germination after treatment with atmospheric pressure cold plasma (APCP) using dielectric barrier discharge (DBD) in seeds that have very low germination rates. To aid industrial applications, several seeds were simultaneously treated with plasma within a space between two coaxial glass tubes sandwiched by two metal mesh screens that produced high-voltage pulses at 17.5kV with a frequency of 990Hz. Three treatment times (3min, 9min and 15min) as well as untreated seeds were used to conduct the wettability, imbibition and germination tests. The wettability and imbibition were found to be directly related to the treatment duration, but saturation of the imbibition was found for treatment durations greater than 9min. Plasma treatment was also effective in improving germination, but shorter treatment duration presented greater germination. This apparent contradiction is explained by the cell damage caused by the increased exposure to plasma, as observed in other studies. The results suggest that there must be an optimal wettability and imbibition condition that ensures that excessive moisture does not harm the germination process.

Biotools: an R function to predict spatial gene diversity via an individual-based approach.

The gene diversity or expected heterozygosity (HE) is based on the allele frequency and is often used as a measure of genetic variability of populations. Knowing the pattern of spatial distribution of HE can be useful for determining strategies of conservation and sampling of collections of individuals. In addition, it can allow one to detect genetic boundaries in a landscape. We adapted a Wombling method based on assignment tests in a circular moving window extensively sampled over the study area in order to estimate HE at points of a prediction grid. The function sHe(), package biotools, is an easy-to-use and flexible implementation in R language that accepts as input geographical and genotyping data. The package biotools is distribution-free under the GPL-2/3 license and currently available from the Comprehensive R Archive Network (CRAN) at . The R platform and all R dependencies are similarly available from CRAN.

Most international guidelines on prevention of healthcare-associated infection lack comprehensive recommendations for neonates and children.

International infection prevention and control (IPC) guidelines provide standardized recommendations for healthcare-associated infection (HCAI) prevention in adults, but often lack specific information about neonates and children. We reviewed ten international IPC/HCAI guidelines to identify paediatric-specific recommendations for HCAI prevention. Hand hygiene, bloodstream infection, ventilator-associated pneumonia, environmental control and outbreak management were frequently reported with recommendations applicable to children and newborns, but documents on catheter-associated urinary tract infection and surgical site infection were lacking.

Estudo preliminar toxicológico, antibacteriano e fitoquímico do extrato etanólico das folhas de Jatropha mollissima (Pohl) Baill. (pinhão-bravo, Euphorbiaceae), coletada no Município de Tauá, Ceará, Nordeste Brasileiro / Toxicological, antibacterial, and phytochemical preliminary study of the ethanolic extract of Jatropha mollissima (Pohl) Baill (pinhão-bravo, Euphorbiaceae) leaves, collected in Tauá, Ceará, Northeastern Brazil

RESUMO A cada dia, cepas bacterianas estão tornando-se resistentes a diversos antibióticos, o que faz necessária a busca de novas substâncias eficazes para o tratamento de doenças. Desta forma, este trabalho reporta o estudo preliminar toxicológico, antibacteriano e fitoquímico do extrato etanólico das folhas de Jatropha mollissima (pinhão-bravo, Euphorbiaceae), coletada no Município de Tauá, Ceará, Nordeste Brasileiro. Inicialmente, realizou-se o teste de toxicidade do extrato contra Artemia salina. Na sequencia, foi realizado o ensaio antibacteriano contra quatro cepas bacterianas Gram-negativas (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883) e uma cepa Gram-positiva (Enterococcus faecalis ATCC 29212). Finalmente, fez-se a análise fitoquímica preliminar do extrato ativo para detecção das principais classes de metabólitos especiais. Como resultado, o extrato etanólico das folhas de J. mollissima se mostrou tóxico para Artemia salina, pois apresentou CL50 igual a 406,02 μg/mL. Quanto à ação antibacteriana, o extrato se mostrou ativo contra a bactéria Gram-positiva Enterococcus faecalis ATCC 29212, apresentando moderada atividade antibacteriana (halo de inibição igual a 7,03 mm). Evidenciou-se no extrato bioativo a presença de cumarinas, fenóis, taninos, flavonoides (flavonóis e flavanonas), alcaloides e esteroides, ambas as classes reportadas como antimicrobianos. Portanto, esse extrato tem potencial para ser usado na produção de fármacos contra infecções causadas por bactérias Gram-positivas. No entanto, as informações direcionam estudos futuros para o isolamento e identificação dos compostos bioativos, monitorados sob a ação antibacteriana mais expressiva.

ABSTRACT Each day, bacterial strains are becoming more resistant to various antibiotics, which requires the search for new effective substances for the treatment of diseases. Thus, this study reports the toxicological, antibacterial, and phytochemical preliminary study of the ethanolic extracts of Jatropha mollissima (pinhão-bravo, Euphorbiaceae) leaves, collected in Tauá, Ceará, Northeast of Brazil. Initially, we performed the toxicity testing of the extract against Artemia salina. Then, we conducted the antibacterial assay against four Gram-negative bacterial strains (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883), and one Gram-positive strain (Enterococcus faecalis ATCC 29212). Finally, we carried out the preliminary phytochemical analysis of the active extract to detect the main classes of special metabolites. As a result, the ethanolic extract of J. mollissima leaves was toxic to Artemia salina, because it presented LC50 equal to 406.02 µg/mL. Regarding antibacterial action, the extract was active against the Gram-positive bacteria Enterococcus faecalis ATCC 29212, with moderate antibacterial activity (inhibition zone equal to 7.03 mm). The bioactive extract had the presence of coumarins, phenols, tannins, flavonoids (flavanols and flavonones), alkaloids and steroids, both classes reported as antimicrobials. Therefore, this extract has the potential to be used in the production of drugs against infections caused by Gram-positive bacteria. However, these information require further studies for the isolation and identification of bioactive compounds, monitored under the more expressive antibacterial action.

Adaptability and stability of genotypes of sweet sorghum by GGEBiplot and Toler methods.

Sweet sorghum has considerable potential for ethanol and energy production. The crop is adaptable and can be grown under a wide range of cultivation conditions in marginal areas; however, studies of phenotypic stability are lacking under tropical conditions. Various methods can be used to assess the stability of the crop. Some of these methods generate the same basic information, whereas others provide additional information on genotype x environment (G x E) interactions and/or a description of the genotypes and environments. In this study, we evaluated the complementarity of two methods, GGEBiplot and Toler, with the aim of achieving more detailed information on G x E interactions and their implications for selection of sweet sorghum genotypes. We used data from 25 sorghum genotypes grown in different environments and evaluated the following traits: flowering (FLOW), green mass yield (GMY), total soluble solids (TSS), and tons of Brix per hectare (TBH). Significant G x E interactions were found for all traits. The most stable genotypes identified with the GGEBiplot method were CMSXS643 for FLOW, CMSXS644 and CMSXS647 for GMY, CMSXS646 and CMSXS637 for TSS, and BRS511 and CMSXSS647 for TBH. Especially for TBH, the genotype BRS511 was classified as doubly desirable by the Toler method; however, unlike the result of the GGEBiplot method, the genotype CMSXS647 was also found to be doubly undesirable. The two analytical methods were complementary and enabled a more reliable identification of adapted and stable genotypes.

Improving the therapeutic potential of endostatin by fusing it with the BAX BH3 death domain.

Endostatin (ES) inhibits angiogenesis, reducing tumor growth in animal models. However, it has low therapeutic effect in human clinical trials. BAX is a member of the BCL-2 family of proteins; its proapoptotic (BH3) domain interacts with other members of the family in the cytoplasm, to induce apoptosis. Here, we fused the BAX BH3 domain with murine ES, to enhance ES potency. Endothelial cells specifically internalize the fusion protein ES-BAX. The presence of the BAX domain enhances endothelial cell death by apoptosis by 1.8-fold and diminishes microvessel outgrowth in the rat aortic ring assay by 6.5-fold. Daily injections of 15 µg of ES-BAX/g in tumor-bearing mice reduce tumor weight by 86.9% as compared with ES-treated animals. Co-immunoprecipitation assays confirmed that ES-BAX interacts with members of the BCL-2 family. Also, ES interacts with BCL-2, BCL-XL, and BAK in endothelial cell lysates, suggesting a potential new mechanism for the apoptosis induction by ES. The superiority of the ES-BAX antiangiogenic effect indicates that this fusion protein could be a promising therapeutic alternative to treat cancer.

Cloning of serine protease cDNAs from Crotalus durissus terrificus venom gland and expression of a functional Gyroxin homologue in COS-7 cells.

Gyroxin is one of main serine proteases of Crotalus durissus terrificus venom, representing about 2% of the protein content in the crude venom. It is a 33 kDa glycoprotein with 3.8% by weight of sugar moiety. This toxin induces hemotoxicity in mice and a neurological condition called barrel rotation syndrome. In the present work, we report the molecular cloning of five new nucleotide sequences from a cDNA library of the venom glands of a single specimen of C. d. terrificus. These sequences have been analyzed in silico with respect to their cDNA organization and similarity with other snake venom serine proteases (SVSPs). We also describe a rapid and efficient method for screening vectors for mammalian cell expression, based on the fact that SVSPs are difficult-to-express toxins due to the presence of several disulfide bonds and glycosylation in their structures. Thus, one of the Gyroxin cDNAs was subcloned into pSectag2 HygroA and pED vectors and used to transfect COS-7 cells. Expression of the functional recombinant Gyroxin isoform was achieved with this cell line with esterase activity in the conditioned culture medium, as revealed by immunoblot of secreted protein and standard anti-crotalic serum from Butantan Institute.

Cloning of a novel acidic phospholipase A2 from the venom gland of Crotalus durissus cascavella (Brazilian northeastern rattlesnake)

The phospholipase A2 superfamily encompasses 15 groups that are classified into: secreted PLA2 (sPLA2); cytosolic PLA2 (cPLA2); Ca2+-independent intracellular PLA2 (iPLA2); platelet-activating factor acetylhydrolase (PAF-AH); and lysosomal PLA2. Currently, approximately 700 PLA2 sequences are known, of which 200 are obtained from the venom gland of Crotalinae snakes. However, thus far, little information is available on cloning, purification and structural characterization of PLA2 from Crotalus durisssus cascavela venom gland. In the present work, we report the molecular cloning of a novel svPLA2 from C. d. cascavella (Cdc), a predominant rattlesnake subspecies in northeastern Brazil. The Cdc svPLA2 cDNA precursor is 689 nucleotides long and encodes a protein of 138 amino acid residues, with a calculated molecular mass of approximately 13,847 Da and an estimated isoelectric point of 5.14. Phylogenetic analysis of Crotalinae PLA2 reveals that Cdc PLA2 clustered with other acidic type IIA PLA2 homologues is also present in the venom of North American rattlesnakes. Hitherto, this study presents a novel PLA2 cDNA precursor from C. d. cascavella and data reported herein will be useful for further steps in svPLA2 purification and analysis.

Expression of mRNAs coding for VAP1/crotastatin-like metalloproteases in the venom glands of three South American pit vipers assessed by quantitative real-time PCR.

Snake venom metalloproteases encompass a large family of toxins, with approximately 200 members already catalogued, which exhibit a diversity of structures and biological functions. From this relatively large number, only a dozen examples of apoptosis-inducing metalloproteases, like VAP1 and 2 from the venom of Crotalus atrox, are known. Since most VAP1-like toxins ever characterized were purified from the venom of Viperidae species inhabiting diverse places on earth, we investigate the expression of VAP-like metalloproteases in the venom gland of three representative pit vipers of the Brazilian territory. By molecular cloning and quantitative real-time polymerase chain reaction, using as calibrator gene the Crotalus durissus terrificus homolog of VAP1, named crotastatin, it is reported here that VAP1/crotastatin-like homologues in the venom gland of Bothrops atrox, C. d. cascavella and Lachesis m. rhombeata are expressed at different levels. Hence, batroxstatins, the crotastatin-like precursors from B. atrox, are expressed 87 times more than crotastatin-1, from C. d. cascavella, and 7.5-fold that lachestatins, from L. m. rhombeata. Moreover, in silico structural analysis of amino acid sequences indicates that batroxstatin-2, crotastatins and lachestatin-1 and -2 which share the archetypal motifs and metal- binding sites of VAP1, are subgrouped in a branch that comprises some apoptosis-inducing toxins.

Identification of crotasin, a crotamine-related gene of Crotalus durissus terrificus.

Crotamine is a cationic peptide (4.9 kDa, pI 9.5) of South American rattlesnake, Crotalus durissus terrificus' venom. Its presence varies according to the subspecies or the geographical locality of a given species. At the genomic level, we observed the presence of 1.8 kb gene, Crt-p1, in crotamine-positive specimens and its absence in crotamine-negative ones. In this work, we described a crotamine-related 2.5 kb gene, crotasin (Cts-p2), isolated from crotamine-negative specimens. Reverse transcription coupled to polymerase chain reaction indicates that Cts-p2 is abundantly expressed in several snake tissues, but scarcely expressed in the venom gland. The genome of crotamine-positive specimen contains both Crt-p1 and Cts-p2 genes. The present data suggest that both crotamine and crotasin have evolved by duplication of a common ancestor gene, and the conservation of their three disulfide bonds indicates that they might adopt the same fold as beta-defensin. The physiological function of the crotasin is not yet known.

Characterization of a Paramyxovirus from a Fer de Lance viper (Bothrops jararaca): partial nucleotide sequence of the putative fusion protein.

During the generation of Expressed Sequence Tags (ESTs) from the Fer de Lance viper (Bothrops jararaca) venom glands, a partial cDNA (clone H8) coding for a protein with all the features of a paramyxovirus fusion protein was characterized. It has 920 bp and codes for a partial protein of 279 amino acids. Two potential N-glycosylation sites are present in the sequence which also possesses a typical membrane anchoring domain made of a stretch of hydrophobic amino acids. The polyadenylation signal sequence was identified. When compared to other fusion proteins, it showed the highest sequence similarity (37-39%) with those of human parainfluenza 3 and Sendai virus.

Cross reactivity of different specific Micrurus antivenom sera with homologous and heterologous snake venoms.

Coral snakes are the only Elapids in America. They are represented by three genera: Leptomicrurus, Micruroides and Micrurus, of which the latter are the most abundant and diversified group. Little is known about the biochemistry of Micrurus venoms due to low availability. Here, we present a study on the cross reactivity of different specific Micrurus antivenom with homologous and heterologous snake venoms in order to contribute to the generation of more efficient antiserum for therapeutic purposes. The three specific antisera tested, anti-Micrurus corallinus, anti-Micrurus frontalis, and anti-Micrurus spixii, as well as the bivalent anti-elapid venom sera, raised against a mixture (50% each) of Micrurus frontalis and Micrurus corallinus venoms, were assayed by Western Blot against Micrurus and non-Micrurus elapid venoms. An antisera raised against a recombinant alpha-neurotoxin-like protein from Micrurus corallinus venom, only reacted in Western blot with its homologous venom, indicating that this protein is specific for Micrurus corallinus coral snake.

[Phlebotominae (Diptera: Psychodidae) of Lagoas, a city of Buriticupu, Amazonia of Maranhão. I. The richness and relative abundance of the species in an area of recent colonization]. / Phlebotominae (Diptera: Psychodidae) de Lagoas, município de Buriticupu, Amazônia Maranhense. I--Riqueza e abundância relativa das espécies em área de colonização recente.

In this study the diversity of the species and relative abundance of the sand flies in the Amazonian community of Lagoas, in Buriticupu, Maranhão State, was determined. The study consisted of adult sandflies captured with CD light-traps, Shannon-trap and manual aspiration tube, between 18 PM and 6 AM, once a month, from December/95 to January/97, in a forest fragment and in the domestic environment (peri and intra). In all, 9,393 specimens were captured (4,302 males and 5,140 females) distributed among 38 species (1 Brumptomyia and 37 Lutzomyia). The diversity and abundance of species was higher in the forest, followed by the peri and intra. The predominance in the wild ambient is a consequence of the potentialities found in the remaining fragments of the forest which have survived the lumber yard exploitation and agricultural activities. In the forest environment Lutzomyia whitmani (64.9%), L. migonei (27%) and L. serrana (3.6%) were the most frequent species. In the peri and intra domicile L. evandroi predominated (55.4% and 97.9%). The species, L. whitmani (33.4%) and L. migonei (6.4%), were important in the peri domicile. The peridomestic habit of 7.7% of the sand flies showed the greatest epidemiologic importance, due to the existence of patients with leishmaniasis ulcers, who believe they were infected near their habitations.

Quantification of crotamine, a small basic myotoxin, in South American rattlesnake (Crotalus durissus terrificus) venom by enzyme-linked immunosorbent assay with parallel-lines analysis.

Intraspecific variation in Crotalus durissus terrificus venom composition was studied in relation to crotamine activity. Crotamine induces paralysis in extension of hind legs of mice and myonecrosis in skeletal muscle cells. To determine whether the venom of crotamine-negative rattlesnake contains a quantity of myotoxin incapable of inducing paralysis, we have developed a very sensitivity immunological assay method, an enzyme-linked immunoabsorbent assay (ELISA), capable of detecting 0.6 ng of purified crotamine. The parallel-lines analysis of ELISA data showed to be useful because it shows the reliability of the experimental conditions. A variation in the amount of myotoxin in the crotamine-positive venom was observed, but not less than 0.1 mg of crotamine per mg of venom. It was not possible to detect it in crotamine-negative venom even at high venom concentrations.

[Comparative study of American tegumentary leishmaniasis between childhood and teenagers from the endemic areas Buriticupu, Maranhao and Corte de Pedra, Bahia, Brazil]. / Estudo comparativo da leishmaniose tegumentar americana em crianças e adolescentes procedentes das áreas endêmicas de Buriticupu (Maranhão) e Corte de Pedra (Bahia), Brasil.

A comparative study on children aged 0-15 years, with American tegumentary leishmaniasis (ATL), in the endemic regions of Buriticupu (MA) and Corte de Pedra (BA), whereby 214 cases were detected between 1982 and 1993, 78 (36.4%) of them originated from Corte de Pedra and 136 (63.6%) from Buriticupu. In Corte de Pedra, most cases were observed in patients aged between 0-5 years. Twenty nine (37.2%) cases; 62% of those were male. In the Buriticupu region, 88 (64.7%) cases occurred in patients of 11-15 years of age, where in 73.8% consisted of male. In both researched regions, brunetts were predominant, with a ratio of 65.4% in Corte de Pedra and 75% in Buriticupu. Twenty six (33.3%) children in the village of Corte de Pedra were farmers, predominantly male (57.7%), generating statistical significance (chi 2 = 11.21). Twenty one (80.8%) farmers were aged 11-15 years. Thirty seven and a half per cent of the children from Buriticupu were students, however, 41 (30.2%) were farmers, representing 39 (44.3%) cases; all of them male. Both in Buriticupu and Corte de Pedra, the unique wound was predominant, corresponding to 57.7% and 53.7%, respectively. The wounding time stood out from one to three months, with 45 (69.2%) cases in Corte de Pedra and 73 (61%) in Buriticupu (chi 2 = 11.82). As to the wound locations, it has been observed that they were most constantly present on the lower limbs, with 77.2% in Corte de Pedra and 58.9% in Buriticupu (chi 2 = 27.9). The cutaneous case mostly found in Corte de Pedra was the ulcerous one (91%). IDRM was positive in 61 (78.2%) children originated from Corte de Pedra, wherein no statistical difference was detected between age ratio and positivity of the test (chi 2 = 0.0669).

Cloning and characterization of a repetitive 1.9 Kb HindIII DNA fragment from Crotalus durissus terrificus genome

Eukaryote genomes are endowed with varying quantities of repeated DNA families. These families show different patterns of conservation among species, copy numbers, chromosomal distribution, and transcription. Characterization of repeated DNA sequences could help to understand the genome anatomy and organization or be used in molecular systematics and molecular evolution studies. We describe here a repetitive DNA sequence of the HindIII family present in the genome of the rattlesnake Crotalus durissus terrificus. In Brazil, the family Crotalus is comprised only by one species durissus, which include several subspecies. The number and distribution of these subspecies are controversial. In the present study, the genomic DNA of a female rattlesnake was digested with HindIII resulting in a strong 1.9 Kb band. A partial genomic library was constructed from the 1.9 Kb DNAs rescued from the agarose gel after HindIII digestion and ligated to the vector pGEM3Zf(+) (Promega). Analysis of 69 clones, 44 hybridized with the 1.9 Kb probe isolated from one of the clones-clone 76, indicating that the DNA isolated from this clone should represent the 1.9 Kb HindIII fragment. This 1.9 Kb HindIII DNA was completely characterized by sequencing.