Inbreeding effects in Solanum lycocarpum A. St.-Hil populations, an endangered species of the Brazilian Cerrado.

The inbreeding effective population size is an estimate of inbreeding and genetic drift in populations. It is an important tool for conservation genetics because it represents the number of individuals that are effectively contributing alleles to the subsequent generations. Several studies have been published in the last decades on the genetic structure of natural plant populations of the Cerrado, the Central-Brazilian savannahs, but most of them do not present effective size estimates. The objective of this study was to show such estimates for Solanum lycocarpum, a Cerrado species that is in danger of genetic erosion. We utilized microsatellites, isozymes, and 2 natural populations for each marker to estimate the population inbreeding effective size of a group of populations (N(^)e(v)) and the minimum number of populations that should be conserved (S(^)(ref)) in order to retain an effective number of 500. For the 2 markers that were utilized, only approximately 12% of the individuals are effective in the populations. The value obtained for S(^)(ref) was approximately 80.

Simple-sequence repeat markers of Cattleya coccinea (Orchidaceae), an endangered species of the Brazilian Atlantic Forest.

Microsatellite markers were developed for the endangered Brazilian orchid species Cattleya coccinea to describe its genetic diversity and structure and to support conservation studies. Nine microsatellite loci were isolated and characterized using an enriched genomic library. All loci are polymorphic at least in the 2 populations sampled, except for loci Cac05 and Cac09 for the Petrópolis population. The mean number of alleles per locus was 8.8 between populations. The mean values of the observed and expected heterozygosities were 0.541 (ranging from 0 to 1) and 0.639 (ranging from 0 to 0.9), respectively. Cross-amplifications were performed in 7 additional Epidendroideae species, and at least 2 loci were successful in 3 additional Cattleya species, Epidendrum secundum, and Brasiliorchis gracilis. All markers described herein will be useful in further studies evaluating the genetic diversity, population dynamics, and conservation genetics of C. coccinea and related species.

Transcriptomic profile of two developmental stages of the cestode parasite Mesocestoides corti.

Cestode development involves complex morphological and physiological changes. Here, we performed a differential expression analysis of gene transcripts between two developmental stages of the model cestode Mesocestoides corti. A RNA-seq-based approach was used to compare the transcriptomes of the tetrathyridium (larval, TT) and strobilated worm (ST) stages of the parasite. We found 19,053 transcripts, from which ∼45% were complete matches to genes previously annotated in the available M. corti draft genome sequence, ∼24% were considered novel isoforms, and ∼24% were considered potential novel transcripts. Stage-specific transcripts were found for both TTs (66) and STs (136), along with shared transcripts significantly overrepresented in one stage (342 in TTs, and 559 in STs). Differential expression and Gene Ontology term enrichment analyzes provided evidence of upregulation of different sets of transcripts associated with 'cytoskeleton', 'metabolism' and 'oxidation-reduction' processes in each stage, suggesting functional involvement of the corresponding genes with stage-specific features. Transcripts and processes enriched in the TT reflect typical larval processes that occur with the parasite in the intermediate host, such as asexual reproduction and budding, as well as active migration from the peritoneum to the liver and vice versa. In STs, transcripts associated with 'development', 'cell growth', and 'morphogenesis' were enriched, along with processes related to sexual reproduction, represented by the upregulation of numerous transcription factors, protein kinases, and histones. Overall, our results contributed to significantly increase the knowledge on the M. corti gene repertoire and expression profile in two developmental stages. Functional implications for the biology of larval and adult cestode parasites and for host-parasite interactions are discussed.

Resin-Based Materials Protect Against Erosion/Abrasion-a Prolonged In Situ Study.

While patient compliance is key to preventive measures related to dental erosion, the application of resin-based materials could serve as an additional treatment to inhibit erosion progression. This in situ study evaluated the effect of applying resin-based materials, including resin infiltrant, on previously eroded enamel subjected to prolonged erosive and abrasive challenges. The factors under study were types of treatment (infiltrant [Icon], sealant [Helioseal Clear], adhesive [Adper Scotchbond Multi-Purpose Plus], and control [no treatment]); wear conditions (erosion [ERO] and erosion + abrasion [ERO + ABR]) and challenge time (5 and 20 days) in a single-phase study. The blocks were prepared from bovine enamel, eroded (0.01 M HCl, pH 2.3 for 30 seconds) and randomized among treatments, wear conditions, and volunteers. The application of resin-based materials followed the manufacturers' recommendations. Twenty-one volunteers wore the palatal intraoral device, in which one row corresponded to ERO and the other to ERO + ABR. In each row, all treatments were represented (2 blocks per treatment). For 20 days, the erosive challenge was performed 4 times/day (immersion in 0.01 M HCl, pH 2.3, for 2 minutes) for the ERO condition. For the ERO + ABR condition, two of the erosive challenges were followed by abrasion for 15 seconds with fluoride dentifrice slurry. Enamel and/or material loss was measured using profilometry (initial, after treatment, and after the end of the fifth and 20th days of in situ erosive challenge) and analyzed by ANOVA models and Tukey's test (α=0.05). The results showed that the application of resin-based materials did not cause superficial enamel loss. The infiltrant group showed a thicker layer of material above the enamel compared with the other materials (p=0.001). After the erosive challenge, there was no difference between the conditions ERO and ERO + ABR (p=0.869). All materials protected the enamel against erosion progression compared with the control group (p=0.001). Based on these results, we conclude that the application of resin-based materials results in protection of previously eroded enamel subjected to in situ erosive and abrasive challenge for 20 days.

Physical localization of the retrotransposons Boudicca and Perere 03 in Schistosoma mansoni.

Schistosoma mansoni is 1 of the causative agents of schistosomiasis, an endemic disease in 76 countries of the world. The study of its genome, estimated to be 270 Mb, is very important to understanding schistosome biology, the mechanisms of drug resistance, and immune evasion. Repetitive elements constitute more than 40% of the S. mansoni genome and may play a role in the parasite evolution. The retrotransposons Boudicca, a long terminal repeat (LTR), and Perere 03, a non-LTR, are present in a high number in the S. mansoni genome and were localized with the use of fluorescence in situ hybridization (FISH) and primed in situ labeling (PRINS). Bacterial artificial chromosomes (BAC) clones containing the retrotransposons Boudicca and Perere 03 were selected by bioinformatic analysis and used as probes in FISH. Using metaphase chromosomes from sporocysts and the FISH and PRINS techniques, we were able to map these retrotransposons. Perere 03 was localized in the euchromatic regions of the short arm of chromosome 2 and Boudicca in the euchromatic regions of the short arm of chromosomes 2 and Z.

Analysis of the cytokine profile in spleen cells from dogs naturally infected by Leishmania chagasi.

Recent studies suggest that asymptomatic dogs infected with canine visceral leishmaniasis (CVL) develop a Th1 immunological profile whilst oligosymptomatic and symptomatic CVL-infected animals present a Th2 profile. In the present study, an RT-PCR method has been standardised and employed to evaluate the frequency and the semi-quantitative level of expression of the cytokines IL-4, IL-10, IL-12, INF-gamma and TNF-alpha in splenocytes of 30 dogs naturally infected with Leishmania chagasi and of 7 non-infected dogs (NID). An increase in the level of expression of IL-12 (p=0.059) was detected in all CVL-infected dogs compared with NID. In dogs exhibiting high parasitism, the frequency of expression of IL-10 was higher (p=0.011) than in animals presenting low parasitism or medium parasitism (MP) and in NID animals, whilst the level of expression of IL-10 was higher (p=0.0094) than in animals exhibiting MP and in the NID group. Positive correlations between the levels of expression of IL-10 with respect to the progression of the disease (IL-10: r=0.3510; p=0.0337) and the levels of expression of IL-10 and INF-gamma increase in parasitism (IL-10: r=0.3428; p=0.0438 and INF-gamma: r=0.4690; p=0.0045) were observed. Such data suggest that CVL is marked by a balanced production of Th1 and Th2 cytokines, with a predominant accumulation of IL-10 as a consequence of an increase in parasitic load and progression of the disease, and INF-gamma was related with the increase in parasitic load.

Corneal astigmatism measuring module for slit lamps.

We have developed an automatic keratometer module for slit lamps that provides automatic measurements of the radii of the corneal curvature. The system projects 72 light spots displayed in a precise circle at the examined cornea. The displacement and deformation of the reflected image of these light spots are analysed providing the keratometry. Measurements in the range of 26.8-75 D can be obtained and a self-calibration system has been specially designed in order to keep the system calibrated. Infrared LEDs indicate automatically which eye is being examined. Volunteer patients (492) have been submitted to the system and the results show that our system has a high correlation factor with the commercially available manual keratometers and the keratometry measurements from a topographer. Our developed system is 95% in agreement with the corneal topographer (Humphrey--Atlas 995 CZM) and the manual keratometer (Topcon OM-4). The system's nominal precision is 0.05 mm for the radii of curvature and 1 degree for the associated axis.

In Vitro Effects of Resin Infiltration on Enamel Erosion Inhibition.

Resin-based materials that show promising effects for preventing the progression of erosion have been studied. This in vitro study evaluated the effects of applying resin-based materials, including resin infiltration, on previously eroded enamel subjected to erosive challenges. The influence of enamel surface etching prior to application of the material was also studied. Bovine enamel blocks were immersed in hydrochloric acid (HCl), 0.01 M (pH 2.3), for 30 seconds in order to form a softened erosion lesion. The blocks were then randomly divided into nine groups (n=12) and treated as follows: C = control without treatment; Hel = pit & fissure resin sealant (Helioseal Clear); Adh = two-step self-etching adhesive system (AdheSe); Tet = two-step conventional adhesive system (Tetric N-bond); and Inf = infiltrant (Icon). The Helno, Adhno, Tetno, and Infno groups received the same materials without (or with no) surface conditioning. The depth of the material's penetration into softened erosion lesions was qualitatively analyzed using reflection and fluorescence confocal microscopy. After application of the materials, the blocks were immersed in HCl for two minutes; this step was followed by immersion in artificial saliva for 120 minutes four times a day for five days (erosive cycling). Both the enamel alteration and material thickness were analyzed using profilometry, and the results were submitted to Kruskal-Wallis and Dunn tests (p>0.05). Images from the confocal microscopy showed minimal penetration of Adh/Adhno and deep penetration of Inf/Infno into the erosive lesions. The groups Hel, Adh, Inf, Tetno, and Infno resulted in the formation of a layer of material over the enamel, which was effective in inhibiting the progression of erosion. In conclusion, the infiltrant, with or without etching, was able to penetrate and protect the enamel against dental erosion. The other resin-based materials, except for the two-step conventional adhesive, were able to penetrate and inhibit the progression of erosive lesions only when they were applied after enamel etching.

Osteossíntese femoral associada ao biovidro 60S em tamanduá-bandeira: relato de caso / Femoral osteosynthesis associated with 60s bioglass in tamanduá bandeira: case report

Atropelamentos de animais silvestres são frequentes e muitas vezes causam fraturas ósseas que, se não tratadas adequadamente, levam à morte do animal por complicações ou por inabilidade de sobrevivência no seu habitat natural. O objetivo do presente trabalho foi relatar o uso de placa associada ao pino intramedular e ao biovidro 60S (BV60S) para o tratamento de fratura de fêmur em tamanduá-bandeira. O animal foi resgatado pela polícia ambiental com suspeita de atropelamento. Foi sedado para avaliação clínica e radiográfica, que revelou fratura em fêmur direito. Utilizou-se dexmedetomidina como medicação pré-anestésica, midazolam e cetamina para indução, e isoflurano para manutenção. Também foi realizado bloqueio peridural com bupivacaína e morfina. A osteossíntese foi feita com placa bloqueada 2,7 e pino intramedular 2,5. Colocaram-se 4g de BV60S no foco de fratura para favorecer a osteogênese. O paciente teve recuperação funcional imediata do membro acometido. A reparação óssea ocorreu por segunda intenção, observando-se ossificação completa do calo com consolidação clínica, aos 30 dias, e remodelação quase completa, aos 180 dias. Conclui-se que o uso de placa e pino associado ao BV60S é eficiente no tratamento de fratura de fêmur em tamanduá, permitindo a rápida recuperação e a reintrodução do animal na natureza.(AU)

Roadblocks of wild animals are frequent and often cause bone fractures that if not properly treated lead to the death of the animal due to complications or inability to survive in its natural habitat. The objective of the present study was to report the use of plate rod and bioglass 60S (BG60S) for the treatment of femoral fracture in anteater. The animal was rescued by environmental police on suspicion of being hit. It was sedated for clinical and radiographic evaluation, which revealed a fracture in the right femur. Dexmedetomidine was used as preanesthetic medication, midazolam and ketamine for induction, and isoflurane for maintenance. Epidural blockade with bupivacaine and morphine was also performed. Osteosynthesis was done with a locking plate 2.7 and 2.5 intramedullary pin. 4G of BG60S was placed in the focus of fracture to favor osteogenesis. The patient had immediate functional recovery of the affected limb. The bone repair occurred by second intention, with complete ossification of the callus with clinical consolidation at 30 days, and near complete remodeling at 180 days. It is concluded that the use of plate rod to the BG60S is efficient in the treatment of femur fracture in anteater, allowing the rapid recovery and reintroduction of the animal in the wild.(AU)

The human immune response to defined immunogens of Schistosoma mansoni: elevated antibody levels to paramyosin in stool-negative individuals from two endemic areas in Brazil.

Sera from individuals living in 2 areas endemic for Schistosoma mansoni in Minas Gerais, Brazil were assayed for the presence of antibodies against paramyosin and glutathione-S-transferase (GST), molecules previously implicated as vaccine immunogens from studies in laboratory hosts. A group was identified consisting of subjects who were stool-negative and had no record of previous infection but who were seropositive by enzyme-linked immunosorbent assay against crude adult worm antigen (SWAP). These individuals had anti-paramyosin antibody levels which were dramatically elevated with respect to those measured in infected (stool-positive) individuals living in the same endemic area. In contrast, the same 2 groups of stool-positive and stool-negative subjects could not be distinguished on the basis of their seroreactivity to either GST or SWAP. After chemotherapy, anti-paramyosin antibodies rose above pre-treatment levels and remained elevated in those individuals who became stool-negative. In contrast, anti-paramyosin antibodies decreased to pretreatment values in drug-treated individuals who failed to show complete parasitological cure. These results suggest that the immune response of humans to paramyosin may play a role in natural resistance to schistosome infection, and that an elevated antibody level against this antigen may be a useful correlate of drug-induced cure.

Sm14 gene expression in different stages of the Schistosoma mansoni life cycle and immunolocalization of the Sm14 protein within the adult worm.

Sm14 is a 14-kDa vaccine candidate antigen from Schistosoma mansoni that seems to be involved in cytoplasmic trafficking of fatty acids. Although schistosomes have a high requirement for lipids, they are not able to synthesize fatty acids and sterols de novo. Thus, they must acquire host lipids. In order to determine whether Sm14 is present in different stages of the life cycle of the parasite, we performed RT-PCR. Sm14 mRNA was identified in all stages of the life cycle studied, mainly schistosomulum, adult worm and egg. Additionally, we used a rabbit anti-Sm14 polyclonal antibody in an indirect immunofluorescence assay to localize Sm14 in adult worm sections. The basal lamella of the tegument and the gut epithelium were strongly labeled. These tissues have a high flow of and demand for lipids, a finding that supports the putative role of Sm14 as an intracellular transporter of fatty acids from host cells.

Hepatitis C virus infection among Brazilian hemophiliacs: a virological, clinical and epidemiological study.

We determined and analyzed risk factors of hepatitis C virus (HCV)-infected Brazilian hemophiliacs according to their virological, clinical and epidemiological characteristics. A cross-sectional and retrospective study of 469 hemophiliacs was carried out at a Brazilian blood center starting in October 1997. The prevalence of HCV infection, HCV genotypes and factors associated with HCV RNA detection was determined. The seroprevalence of anti-HCV antibodies (ELISA-3.0) was 44.6% (209/469). Virological, clinical and epidemiological assessments were completed for 162 positive patients. There were seven (4.3%) anti-HCV seroconversions between October 1992 and October 1997. During the same period, 40.8% of the positive anti-HCV hemophiliacs had abnormal alanine transaminase (ALT) levels. Plasma HCV RNA was detected by nested-RT-PCR in 116 patients (71.6%). RFLP analysis showed the following genotype distribution: HCV-1 in 98 hemophiliacs (84.5%), HCV-3 in ten (8.6%), HCV-4 in three (2.6%), HCV-2 in one (0.9%), and not typeable in four cases (3.4%). Univariate analysis indicated that older age (P = 0.017) and abnormal ALT levels (P = 0.010) were associated with HCV viremia, while the presence of inhibitor antibodies (P = 0.024) and HBsAg (P = 0.007) represented a protective factor against the presence of HCV RNA. These findings may contribute to a better understanding of the relationship between HCV infection and hemophilia.

Physicochemical characterization and antioxidant activity of decolorized Moringa oleifera Lam leaf flour

BACKGROUND: Maringa oleifera leaves are rich in antioxidant substances; however, when lyophilized leaves were used in flour form in meat products, they presented no antioxidant effect and even accelerated the oxidation process of the product. Thus, the objective of this study was to evaluate the effect of chlorophyll extraction on the physicochemical composition and antioxidant activity of Moringa leaves. METHODS: Moringa leaves were dried and ground in order to obtain uniform flour. A treatment using chlorophyll extraction (decolorized) was tested versus a control treatment(non-decolorized) for proximate composition, instrumental color, and antioxidant activity using ANOVA followed by Tukey'stest. RESULTS: Higher crude fiber, ash, and protein contents were observed for decolorized flour (19.41 and 38.13%, 11.87 and14.02%, and 28.81 and 31.33%, respectively) when compared to those for the control. Chlorophyll extraction significantly affected (p< 0.05) the instrumental color of the leaves flour. The half maximal effective concentration (EC50) of both decolorized and control flour was 3.74 and 4.30 mg/L, respectively. The equivalent of antioxidant per gramof non-decolorized leaves was higher than that observed for the decolorized leaves (0.36 and 0.32 g/g DPPH, respectively). The antioxidant activity (AA%) of the extract from non-decolorized leaves was higher in the concentrations of 5 and 2.5 mg/0.1 mL, while the decolorized leaves was higher in the extract concentration 5 and 2 mg/0.1 ml. CONCLUSION: The decolorization process affected the chemical composition and color of Moringa oleifera leaves flours however did not improve its antioxidant activity

Campomanesia lineatifolia Ruiz & Pav. as a gastroprotective agent.

ETHNOPHARMACOLOGICAL RELEVANCE: Campomanesia lineatifolia Ruiz and Pav. (Myrtaceae) is a native edible species found in the Amazon Rainforest, commonly known as gabiroba. In Brazil, Campomanesia species are frequently used in traditional medicine for gastrointestinal disorders. MATERIALS AND METHODS: The present study performed phytochemical analyses and determined both the in vitro antioxidant activity of the ethanolic extract of Campomanesia lineatifolia leaves (EEC) as well as its ethyl acetate fraction (EAFC). In this analysis, quercetin was used as a positive control. Gastroprotective activity was also investigated at different oral doses in two experimental models in rats - gastric lesion induced by ethanol and gastric lesion induced by indomethacin. In this analysis, cimetidine and sucralfate were used as positive controls. The area of gastric lesion underwent macroscopic and histomorphometric evaluations, while the mucus content was estimated by applying the periodic acid-Schiff stain. Oral acute toxicity was also assessed. RESULTS: Phytochemical studies revealed the presence of flavonoids and tannins. Catechin and quercitrin were isolated by bioguided chromatographic fractionation of EAFC. EEC and EAFC presented in vitro antioxidant activity. The oral administration of EEC and EAFC at doses 100-400 mg/kg (ethanol model) and at doses of 400-1200 mg/kg (indomethacin model) proved to be effective in preventing gastric ulcerations in rats. Pretreatment with EAFC (400mg/kg) significantly increased the gastric mucus content in the ethanol model. No animals died during the acute oral toxicology test. CONCLUSIONS: Results confirm the Brazilian ethnopharmacological use of Campomanesia lineatifolia as a gastroprotective agent and indicate that the anti-ulcer effect is most likely mediated by scavenging free radicals due to the polyphenol content and, at least in part, by increasing the mucus secretion and the mucosal defense. In addition, EEC and EAFC were found to be safe when applied to a 2000 mg/kg single oral dose.

Squamous cell abnormalities in exfoliated cells from the urine of Schistosoma haematobium-infected adults in a rural fishing community in Nigeria.

Schistosoma haematobium infection is endemic in Nigeria, with substantial transmissions in all the states of the federation and a high prevalence rate in schools. Literature has linked bladder cancer, mostly squamous cell type, with long-term S. haematobium infections. The objective of this descriptive study was to screen exfoliated cells in the urine of S. haematobium-infected patients for squamous cell abnormalities through cytopathological examinations. Study participants were drawn from Imala Odo, a community near Oyan Dam in Abeokuta North Local Government Area, Ogun state, Southwest Nigeria. Due to a considerable day-to-day variation of S. haematobium eggs in urine, 3 rounds of 200 ml of urine samples were collected on 3 different days from 32 infected patients and 10 uninfected controls and examined. Cytological preparations of the infected 15 males and 8 females and 10 controls (5 males and 5 females) were screened for squamous cell abnormalities. Severely dysplastic to frankly malignant squamous cells were observed in 1 (3.1%) male and 2 (6.3%) females, while no abnormality was observed in the controls.