What Is the Microbial Profile in Persistent Endodontic Infections? A Scoping Review.

INTRODUCTION: This scoping review aimed to map the evidence about the microbiota found in persistent endodontic infections. METHODS: The study protocol was prospectively registered and is available at https://osf.io/3g2cp. The electronic search was performed in MEDLINE via PubMed, Lilacs, BBO, Scopus, Web of Science, Cochrane Library, and Embase. The eligibility criteria were based on the PCC acronym, where P (Population) represents patients with teeth presenting persistent endodontic infection, C (Concept) represents microbial profile, and C (Context) represents undergoing endodontic retreatment. Clinical studies that evaluated the microbial profile of samples collected from root canals of teeth undergoing retreatment, using classical or molecular methods, were included. Studies that did not show a minimum period of 1 year between primary endodontic treatment and retreatment or did not radiographically evaluate the quality of primary root canal filling were excluded. Two reviewers independently selected the articles and collected data. RESULTS: From a total of 957 articles, 161 were read in full, and 32 studies were included. The most prevalent species were Enterococcus faecalis, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Dialister invisus, Propionibacterium acnes, Tannerella forsythia, and Treponema denticola. Cases with symptomatology or inadequate root canal filling presented an increase in specific bacterial species compared to those with no symptomatology or adequate filling. A greater number of microorganisms was observed in teeth with inadequate coronal restoration compared to those with adequate restoration. CONCLUSIONS: Persistent endodontic infections have a polymicrobial profile identified by the commonly used methods for bacterial detection/identification and are subject to the limitations present in each of those methods.

Influence of variations in the environmental pH on the solubility and water sorption of a calcium silicate-based root canal sealer.

AIM: To evaluate the influence of pH variation on the solubility and water sorption of a premixed calcium silicate-based root canal sealer (EndoSequence BC Sealer) compared to the gold standard based on epoxy resin (AH Plus Jet) after immersion in distilled water and phosphate-buffered saline (PBS). METHODOLOGY: Solubility and water sorption were evaluated after immersion in distilled water or PBS at several pHs (5, 7 and 12) and the values were calculated as percentages of the original mass after 24 h, 7 and 30 days of immersion. The crystalline structures present in the sealers and surface precipitates were assessed by X-ray diffraction. The Shapiro-Wilk's test revealed that data were normally distributed; thus, statistical analysis was performed using one-way anova and Tukey's tests or independent t-test, assuming a 5% α-error. RESULTS: EndoSequence BC Sealer was associated with significantly greater water sorption and solubility compared to AH Plus Jet in all tested conditions (P < 0.05). The acid environment increased the 24 h solubility of EndoSequence BC Sealer immersed in PBS (P < 0.05) and did not induce significative changes in the water sorption (P > 0.05). Alkaline pH reduced the solubility of EndoSequence BC Sealer and increased that of AH Plus Jet at all experimental times and soaking media (P < 0.05). Alkaline environment also significantly increased the water sorption of AH Plus Jet immersed in PBS (P < 0.05). Immersion in PBS significantly reduced the solubility of EndoSequence BC Sealer and significantly increased that of AH Plus Jet (P < 0.05). Precipitates on the surface of EndoSequence BC Sealer corresponding to hydroxyapatite and calcium carbonate were detected after immersion in PBS at pH 5 and 7 for 30 days. CONCLUSIONS: EndoSequence BC Sealer had significantly greater solubility and water sorption than AH Plus Jet. Although the alkaline pH and soaking media directly influenced the solubility and water sorption of the sealers, the solubility of AH Plus Jet remained within the limits recommended by ISO 6876, whilst the solubility of the EndoSequence BC Sealer did not comply with ISO recommendations in all the conditions tested.

Influence of ultraconservative access cavities on instrumentation efficacy with XP-endo Shaper and Reciproc, filling ability and load capacity of mandibular molars subjected to thermomechanical cycling.

AIM: To evaluate the influence of ultraconservative access cavities (UltraAC) on canal shaping and filling ability and load capacity of mandibular molars after root canal preparation with XP-endo Shaper (XP) or Reciproc (RC), under simulated clinical conditions. Traditional access cavities (TradAC) were used for comparison. METHODOLOGY: Forty extracted mandibular molars were scanned by micro-computed tomographic and, based on similar anatomical features, were divided into four groups (n = 10), according to the type of access cavity and canal instrumentation protocol: TradAC/RC, TradAC/XP, UltraAC/RC and UltraAC/XP. All root canal procedures were performed with the teeth placed in a dental mannequin. Teeth were scanned after root canal instrumentation and filling. Unprepared canal area, dentine removed, accumulated hard tissue debris (AHTD), canal transportation, presence of voids and filling material remnants within the pulp chamber were analysed. After restorative procedures, the teeth were subjected to thermomechanical cycling and to a load capacity test. Statistical analyses were performed using two-way anova test, considering the randomized blocks design (P < 0.05). RESULTS: The percentage of unprepared canal area was significantly lower in TradAC groups when compared to UltraAC groups (P < 0.05), regardless of the instrument used. The UltraAC/XP group had significantly lower percentage of root dentine removed when compared to other groups (P < 0.05). UltraAC/XP and TradAC/XP groups had significantly lower percentages of AHTD than UltraAC/RC and TradAC/RC groups (P < 0.05). Regarding canal transportation, in the MB root canals, the TradAC/XP group had significantly lower values than other groups (P < 0.05). In general, in ML and distal root canals, TradAC/XP and UltraAC/XP groups had significantly lower values of canal transportation when compared to other groups (P < 0.05). Moreover, the UltraAC/RC had the greatest canal transportation values in MB and distal canals. The UltraAC groups had significantly greater percentages of voids and volume of remaining root filling material within the pulp chamber after cleaning procedures than TradAC groups (P < 0.05). There was no difference in the load capacity amongst groups (P < 0.05). CONCLUSION: The UltraAC/XP and UltraAC/RC groups had significantly greater areas of unprepared canal walls, significantly more voids and volume of root filling material remnants within the pulp chamber after canal filling. UltraAC/XP was associated with significantly less root dentine removal and significantly more AHTD whilst TradAC/XP had overall significantly less canal transportation. No differences were observed in the load capacity amongst groups.

Preserving dentine in minimally invasive access cavities does not strength fracture resistance of restored mandibular molars.

AIM: To evaluate the correlation between the volume of dental hard tissues removed and the fracture resistance of mandibular molar teeth with ultraconservative (UltraAC) or traditional (TradAC) access cavity preparations. METHODOLOGY: Sixty recently extracted and intact mandibular first molars were scanned in a microcomputed tomographic (micro-CT) device, anatomically matched and assigned at random to 2 groups (n = 30), according to the access cavity type: UltraAC or TradAC. After access preparation, mesial and distal canals were prepared using Reciproc instruments and a new scan was taken. The volumes of pulp chamber and dental hard tissues in each specimen were measured before and after the experimental procedures, and the percentage volume reduction of hard tissues calculated for the entire tooth and for the crown separately. Teeth were then root filled, restored and subjected to fracture resistance tests in an universal testing machine. The force required to fracture was recorded in Newtons (N). The adherence of the variables to a Gaussian curve was verified using a Shapiro-Wilk test. Skewed data were analysed with nonparametric Wilcoxon signed rank or Spearman correlation tests, whilst the normally distributed data were analysed with a dependent samples Student t-test. Level of significance was set at 5%. RESULTS: Pre-operatively, no difference was observed between groups regarding the volume of pulp canal space or dental hard tissues (P > 0.05). After access preparation, volumes of the removed hard tissues from the entire tooth and from the crown were significantly greater in teeth prepared with TradAC than with UltraAC (P < 0.05). No significant difference was observed in the fracture resistance values between TradAC (902.9 ± 347.8 N) and UltraAC (948.7 ± 405.7 N) groups (P = 0.975). The Spearman test was unable to identify a correlation between fracture resistance and percentage volume of removed dental hard tissues in the entire root (P = 0.525, r = -0.084) or only in the crown (P = 0.152, r = -0.187). CONCLUSION: The volume of hard dental tissues removed, although greater in teeth with TradAC compared to UltraAC, did not correlate with the fracture resistance results, indicating that a minimally invasive access cavity did not increase the resistance of restored mandibular first molars to fracture.

Performance and Meat Quality of Broiler Chickens Reared on two Different Litter Materials and at two Stocking Densities.

1. This study evaluated the performance and meat quality of broiler chickens reared on two litter materials and at two stocking densities.2. The chicks were allotted in a completely randomised design in a 2 × 2 factorial arrangement with two litter materials (wood shavings or rice straw) and two stocking densities (24 or 30 kg/m2), with six replicates per treatment. Broiler performance, carcase yield, prime cuts, blood plasma proteins, the enzymes alanine aminotransferase and aspartate aminotransferase, edible viscera, immune organs, intestine weight and length, abdominal fat, breast meat colour, pH, weight loss by defrosting, weight loss by cooking and shear force were evaluated.3. The density of 24 kg/m2 positively influenced feed consumption and weight gain. The type of litter had a positive influence on feed conversion, with a lower value for birds raised on wood shavings.4. Breast production was improved in birds reared at a density of 24 kg/m2 when compared to birds reared at a density of 30 kg/m2. The rice straw bedding negatively affected abdominal fat (resulting in higher percentages) when compared to birds reared on wood shavings.5. The types of litter affected the enzyme aspartate aminotransferase in birds raised on wood shavings, although these values were not increased to the point of influence on the physiological functions of broilers.6. The colour of the breast meat, pH, temperature, weight loss after thawing or cooking, shear force, cholesterol, triglycerides, albumin, total proteins, glucose or alanine aminotransferase content were not influenced by the stocking densities or bedding.7. The use of wood shavings as poultry bedding at a stocking density of 24 kg/m2resulted in the best performance for broiler chickens at 42 days of age.

The influence of endodontic access cavity design on the efficacy of canal instrumentation, microbial reduction, root canal filling and fracture resistance in mandibular molars.

AIM: To assess the impact of conservative endodontic access cavities (CEC) and truss access cavities (TAC) during root canal treatment performed on mandibular molars in terms of: ability to shape and fill root canals, microbial reduction in canals, and cleaning of the pulp chamber. In addition, the fracture resistance of the teeth after coronal restoration was assessed. Traditional endodontic cavities (TEC) were used as a reference technique for comparison. METHODOLOGY: Thirty extracted intact mandibular molars were scanned in a microcomputed tomography device (micro-CT), matched based on similar anatomical features and assigned to TEC, CEC or TAC groups (n = 10). The specimens were accessed accordingly, and root canals were contaminated with bacterial suspensions of Enterococcus faecalis (21 days). Subsequently, the first microbial sample was collected from root canals (S1). The canals were initially prepared with Reciproc Blue R25 instrument followed by a second instrumentation using Reciproc Blue R40. Eight mL of 0.5% NaOCl were used as an irrigant for each instrument. A final irrigation protocol was performed with 2 mL of 0.5% NaOCl, 2 mL of 17% EDTA and another 2 mL of 0.5% NaOCl. Microbial samples were collected from root canals after R25 (S2), R40 (S3) and final irrigation (S4). The teeth were rescanned after S4. Then, root canals were filled, rescanned, restored and the teeth subjected to fracture resistance tests. The statistical analysis was performed with type I negative binomial and beta 0-1 inflation regression models for microbiological analysis. Instrumentation, filling and resistance to fracture results were subjected to anova and Tukey tests (P < 0.05). RESULTS: S4 revealed no significant variations in microbial reduction amongst the groups (P > 0.05). TEC had a significantly lower percentage of unprepared surface area than CEC (P < 0.05). No differences were found regarding the percentage of dentine removed, transportation, centring ability and filling voids amongst the groups (P > 0.05). The TEC group had a significantly lower volume of remaining root filling material within the pulp chamber than CEC and TAC groups (P < 0.05). There was no difference regarding fracture resistance amongst the groups (P > 0.05). CONCLUSION: Conservative access cavities did not offer any advantage in comparison with the traditional endodontic cavities in any of the parameters considered. Furthermore, conservative methods were associated with larger percentages of unprepared canal surface area and larger volumes of remaining root filling material within the pulp chamber.

The impact of minimally invasive root canal preparation strategies on the ability to shape root canals of mandibular molars.

AIM: To evaluate the use of a reduced taper endodontic instrument system (Bassi Logic™ .03 taper) and expandable heat-treated system (XP-endo Shaper) on the ability to shape canals in mandibular molars, by means of microcomputed tomography (micro-CT) analysis. The Reciproc system was used as the reference for comparison. METHODOLOGY: Twenty-four mandibular molars were scanned in a micro-CT, matched based on similar anatomical features and sorted into three groups (n = 8 per group), according to root canal preparation system: Bassi Logic™ .03, XP-endo Shaper and Reciproc. The teeth were mounted onto a mannequin and the pulp chambers were accessed with traditional access cavities. In Bassi Logic™ .03 and Reciproc groups, mesial canals were prepared with size 25, .03 taper or R25 (size 25, .08v taper) instruments and the distal root canal with size 25, .03 taper and size 40, .03 taper or R25 and R40 (size 40, .06v taper), respectively. In XP-endo Shaper groups, all root canals were prepared with XP-endo Shaper (size 30, .04v taper). After root canal preparation, the teeth were rescanned. The percentage of untouched canal areas and the percentage of removed dentine were evaluated separately for mesial and distal root canals. The data were analysed using one-way anova and Tukey tests (P < 0.05). RESULTS: The Bassi Logic™ .03 group was associated with a greater percentage of untouched canal areas when compared to XP-endo Shaper and Reciproc groups for mesial and distal root canals (P < 0.05), but no differences were found between XP-endo Shaper and Reciproc groups (P > 0.05). No significant difference was observed in the percentage of dentine removed amongst the groups for mesial and distal root canals (P > 0.05). CONCLUSION: The use of a reduced taper system (Bassi Logic™ .03) during root canal preparation resulted in a greater percentage of untouched canal areas when compared to XP-endo Shaper and Reciproc instruments. No differences were observed amongst the systems regarding the percentage of dentine removed.

A laboratory study of the impact of ultraconservative access cavities and minimal root canal tapers on the ability to shape canals in extracted mandibular molars and their fracture resistance.

AIM: To evaluate the influence of ultraconservative endodontic access cavities (UEC) and the use of instruments with various tapers (.03 or .05) and tip diameters (25 or 40) on the ability to shape canals in mandibular molars and their fracture resistance in comparison with traditional endodontic access cavities (TEC). METHODOLOGY: Thirty-two intact extracted mandibular molars were scanned in a micro-computed tomography device, matched based on similar anatomical features and assigned to UEC or TEC groups and to subgroups according to mechanical canal preparation with Bassi Logic™ rotary NiTi instruments with .03 or .05 taper (n = 8 per group). The teeth were mounted onto a mannequin head, and the pulp chambers were accessed and the root canals subjected to instrumentation. In the first stage, size 25 endodontic instruments were used, with .03 or .05 tapers. The teeth were re-scanned. Subsequently, a second canal instrumentation was performed with the size 40 instruments with .03 or .05 tapers and scanned again. The root canals were filled and then the teeth restored before being subjected to fracture resistance tests. The data on untouched canal areas, canal transportation, centring ability, the percentage of removed dentine and maximum load at fracture were analysed. One-way anova and Tukey's tests were performed for inter-group analysis. The paired-samples t-test was performed on the results between size 25 and size 40 apical preparations for each individual specimen (P < 0.05). RESULTS: No significant difference was observed for the percentage of untouched canal area when comparing UEC and TEC groups or between the .03 and .05 tapers with the same instrument tip size (P > 0.05). There was a significantly lower percentage of untouched canal area after instrumentation with size 40 instruments when compared to size 25 (P < 0.05). No significant difference in the percentage of dentine removed was observed between the access cavities or the different tapers (P > 0.05). Size 40 apical preparations removed significantly more dentine than size 25 preparations (P < 0.05). No significant differences were observed in canal transportation, centring ability and fracture resistance in any of the tested groups (P > 0.05). CONCLUSIONS: Ultraconservative endodontic access cavities did not offer any advantages in comparison with traditional endodontic access cavities on the ability to shape canals, nor the fracture resistance of mandibular molars. No differences were observed in shaping ability and fracture resistance between .03 and .05 taper canal preparations. Apical preparation with larger instruments resulted in significantly less untouched canal area in all groups.

Calcium chloride-enriched calcium aluminate cement promotes in vitro osteogenesis.

AIM: To evaluate the effects of 2.8% or 10% calcium chloride (CaCl2 ) in calcium aluminate cement (CAC) with either bismuth oxide (Bi2 O3 ) or zinc oxide (ZnO) as radiopacifiers on the progression of osteogenic cell cultures. METHODOLOGY: Rat calvaria-derived cells were grown on Thermanox® coverslips for 24 h and exposed to samples of (i) CACb: with 2.8% CaCl2 and 25% Bi2 O3 ; (ii) CACb+: with 10% CaCl2 and 25% Bi2 O3 ; (iii) CACz: with 2.8% CaCl2 and 25% ZnO; or (iv) CACz+: with 10% CaCl2 and 25% ZnO, placed on inserts. Nonexposed cultures served as the control. Calcium and phosphorus contents in culture media were quantified. The effects of the cements on cell apoptosis, cell viability and acquisition of the osteogenic cell phenotype were evaluated. Data were compared by Kruskal-Wallis test (α = 5%). RESULTS: CACb+ promoted the highest levels of calcium in the culture media; CACz+, the lowest levels of phosphorus (P < 0.05). CACz+ and CACb increased cell apoptosis (P < 0.05). CACb reduced cell viability (P < 0.05) and the expression of the osteoblastic phenotype. CACz+ and CACb+ promoted greater cell differentiation and matrix mineralization compared to CACz and CACb (P < 0.05). CONCLUSION: For CAC with the lower CaCl2 content, the use of Bi2 O3 was detrimental for osteoblastic cell survival and differentiation compared to ZnO, while CAC with the higher CaCl2 content supported the acquisition of the osteogenic cell phenotype in vitro regardless of the radiopacifier used. Thus, CAC with 10% CaCl2 would potentially promote bone repair in the context of endodontic therapies.

Measurement of the proton-air cross section at √s=57 TeV with the Pierre Auger Observatory.

We report a measurement of the proton-air cross section for particle production at the center-of-mass energy per nucleon of 57 TeV. This is derived from the distribution of the depths of shower maxima observed with the Pierre Auger Observatory: systematic uncertainties are studied in detail. Analyzing the tail of the distribution of the shower maxima, a proton-air cross section of [505±22(stat)(-36)(+28)(syst)] mb is found.

Indication of reactor ν(e) disappearance in the Double Chooz experiment.

The Double Chooz experiment presents an indication of reactor electron antineutrino disappearance consistent with neutrino oscillations. An observed-to-predicted ratio of events of 0.944±0.016(stat)±0.040(syst) was obtained in 101 days of running at the Chooz nuclear power plant in France, with two 4.25 GW(th) reactors. The results were obtained from a single 10 m(3) fiducial volume detector located 1050 m from the two reactor cores. The reactor antineutrino flux prediction used the Bugey4 flux measurement after correction for differences in core composition. The deficit can be interpreted as an indication of a nonzero value of the still unmeasured neutrino mixing parameter sin(2)2θ(13). Analyzing both the rate of the prompt positrons and their energy spectrum, we find sin(2)2θ(13)=0.086±0.041(stat)±0.030(syst), or, at 90% C.L., 0.017

Measurement of the depth of maximum of extensive air showers above 10{18} eV.

We describe the measurement of the depth of maximum, X{max}, of the longitudinal development of air showers induced by cosmic rays. Almost 4000 events above 10;{18} eV observed by the fluorescence detector of the Pierre Auger Observatory in coincidence with at least one surface detector station are selected for the analysis. The average shower maximum was found to evolve with energy at a rate of (106{-21}{+35}) g/cm{2}/decade below 10{18.24+/-0.05} eV, and (24+/-3) g/cm{2}/decade above this energy. The measured shower-to-shower fluctuations decrease from about 55 to 26 g/cm{2}. The interpretation of these results in terms of the cosmic ray mass composition is briefly discussed.

A 245kb mini-chromosome impacts on Leishmania braziliensis infection and survival.

Leishmania (V.) braziliensis, the causative agent of mucocutaneous leishmaniasis in the New World, may present an LD1 type genomic amplification that appears as a small 245 kb linear chromosome, and is not clearly associated to the presence of a selection agent. A bt1 gene, codifying for a biopterin transporter protein, was identified in this small chromosome. Leishmania are auxotrophic for pterins and one of the proposed explanations for the appearance of this amplification is the improvement of biopterin capture by the parasite. We analyzed some biological aspects of two lineages of L. braziliensis strain M2903, with and without the small amplified chromosome. We showed differences in infectivity of these lineages, in macrophages and the insect vector Lutzomyia longipalpis, as well as in the uptake and metabolization of intermediates of the Leishmania biopterin salvage pathway. Our results suggest that the genomic amplification favors survival due to improved biopterin capture and at the same time hinders the infective capability, suggesting that within a population different parasites can perform different roles.

Observation of the suppression of the flux of cosmic rays above 4 x 10 (19) eV.

The energy spectrum of cosmic rays above 2.5 x 10;{18} eV, derived from 20,000 events recorded at the Pierre Auger Observatory, is described. The spectral index gamma of the particle flux, J proportional, variantE;{-gamma}, at energies between 4 x 10;{18} eV and 4 x 10;{19} eV is 2.69+/-0.02(stat)+/-0.06(syst), steepening to 4.2+/-0.4(stat)+/-0.06(syst) at higher energies. The hypothesis of a single power law is rejected with a significance greater than 6 standard deviations. The data are consistent with the prediction by Greisen and by Zatsepin and Kuz'min.

Upper limit on the diffuse flux of ultrahigh energy tau neutrinos from the Pierre Auger Observatory.

The surface detector array of the Pierre Auger Observatory is sensitive to Earth-skimming tau neutrinos that interact in Earth's crust. Tau leptons from nu(tau) charged-current interactions can emerge and decay in the atmosphere to produce a nearly horizontal shower with a significant electromagnetic component. The data collected between 1 January 2004 and 31 August 2007 are used to place an upper limit on the diffuse flux of nu(tau) at EeV energies. Assuming an E(nu)(-2) differential energy spectrum the limit set at 90% C.L. is E(nu)(2)dN(nu)(tau)/dE(nu)<1.3 x 10(-7) GeV cm(-2) s(-1) sr(-1) in the energy range 2 x 10(17) eV< E(nu)< 2 x 10(19) eV.

Correlation of the highest-energy cosmic rays with nearby extragalactic objects.

Using data collected at the Pierre Auger Observatory during the past 3.7 years, we demonstrated a correlation between the arrival directions of cosmic rays with energy above 6 x 10(19) electron volts and the positions of active galactic nuclei (AGN) lying within approximately 75 megaparsecs. We rejected the hypothesis of an isotropic distribution of these cosmic rays with at least a 99% confidence level from a prescribed a priori test. The correlation we observed is compatible with the hypothesis that the highest-energy particles originate from nearby extragalactic sources whose flux has not been substantially reduced by interaction with the cosmic background radiation. AGN or objects having a similar spatial distribution are possible sources.

Characterization of constitutive and putative differentially expressed mRNAs by means of expressed sequence tags, differential display reverse transcriptase-PCR and randomly amplified polymorphic DNA-PCR from the sand fly vector Lutzomyia longipalpis.

Molecular studies of insect disease vectors are of paramount importance for understanding parasite-vector relationship. Advances in this area have led to important findings regarding changes in vectors' physiology upon blood feeding and parasite infection. Mechanisms for interfering with the vectorial capacity of insects responsible for the transmission of diseases such as malaria, Chagas disease and dengue fever are being devised with the ultimate goal of developing transgenic insects. A primary necessity for this goal is information on gene expression and control in the target insect. Our group is investigating molecular aspects of the interaction between Leishmania parasites and Lutzomyia sand flies. As an initial step in our studies we have used random sequencing of cDNA clones from two expression libraries made from head/thorax and abdomen of sugar fed L. longipalpis for the identification of expressed sequence tags (EST). We applied differential display reverse transcriptase-PCR and randomly amplified polymorphic DNA-PCR to characterize differentially expressed mRNA from sugar and blood fed insects, and, in one case, from a L. (V.) braziliensis-infected L. longipalpis. We identified 37 cDNAs that have shown homology to known sequences from GeneBank. Of these, 32 cDNAs code for constitutive proteins such as zinc finger protein, glutamine synthetase, G binding protein, ubiquitin conjugating enzyme. Three are putative differentially expressed cDNAs from blood fed and Leishmania-infected midgut, a chitinase, a V-ATPase and a MAP kinase. Finally, two sequences are homologous to Drosophila melanogaster gene products recently discovered through the Drosophila genome initiative.

Characterization of constitutive and putative differentially expressed mRNAs by means of expressed sequence tags, differential display reverse transcriptase-PCR and randomly amplified polymorphic DNA-PCR from the sand fly vector Lutzomyia longipalpis

Molecular studies of insect disease vectors are of paramount importance for understanding parasite-vector relationship. Advances in this area have led to important findings regarding changes in vectors' physiology upon blood feeding and parasite infection. Mechanisms for interfering with the vectorial capacity of insects responsible for the transmission of diseases such as malaria, Chagas disease and dengue fever are being devised with the ultimate goal of developing transgenic insects. A primary necessity for this goal is information on gene expression and control in the target insect. Our group is investigating molecular aspects of the interaction between Leishmania parasites and Lutzomyia sand flies. As an initial step in our studies we have used random sequencing of cDNA clones from two expression libraries made from head/thorax and abdomen of sugar fed L. longipalpis for the identification of expressed sequence tags (EST). We applied differential display reverse transcriptase-PCR and randomly amplified polymorphic DNA-PCR to characterize differentially expressed mRNA from sugar and blood fed insects, and, in one case, from a L. (V.) braziliensis-infected L. longipalpis. We identified 37 cDNAs that have shown homology to known sequences from GeneBank. Of these, 32 cDNAs code for constitutive proteins such as zinc finger protein, glutamine synthetase, G binding protein, ubiquitin conjugating enzyme. Three are putative differentially expressed cDNAs from blood fed and Leishmania-infected midgut, a chitinase, a V-ATPase and a MAP kinase. Finally, two sequences are homologous to Drosophila melanogaster gene products recently discovered through the Drosophila genome initiative

[Acquired immunodeficiency syndrome in children: pathogenesis, prognostic markers, and treatment] / Síndrome da imunodeficiência adquirida na criança. Aspectos da patogênese, marcadores prognósticos e tratamento.

OBJECTIVE: To review recent knowledge about Acquired Immunodeficiency Syndrome in children to help pediatricians provide care for these patients.METHOD: We reviewed the most recent papers about pathogenesis, prognostic markers and treatment of the HIV- infected child.RESULTS: We present the main HIV features that may interfere in the pathogenesis of this syndrome and that should be taken into consideration when the antiretroviral therapy starts.CONCLUSIONS: Viral replication in children is very intense, and the persistence of high levels of viral load may be a sign of immunological immaturity. Viral load measurement and CD4+ lymphocytes count are important markers of the disease progression. At least two drugs should be administered as antiretroviral therapy, which should aim at reducing the viral load to undetectable levels. This will slow down the progression of the disease and the emergence of resistant viral strains.