Larval trematodes hosted by Biomphalaria straminea in the Brazilian semiarid region: implications for schistosomiasis control.

The present study aimed to identify larval trematodes shed by snails found in water bodies used by urban communities in a former schistosomiasis endemic area in the state of Piauí, in the Brazilian semiarid region. A malacological survey was performed followed by analysis of the cercariae shed by the snails after light exposure. Biomphalaria straminea specimens (n=1,224) were obtained from all seven collection sites. Cercariae shed by snails were i) single tailed, in which one type of cercariae was identified ( Echinostoma cercariae), and ii) with bifurcated tail (brevifurcate apharyngeate distome, brevifurcate pharyngeate distome, and longifurcate pharyngeate distome [strigeocercaria]). Brevifurcate apharyngeate distome were further examined and the presence of spikes in swimming membranes enabled the identification of Spirorchiidae cercariae in all individuals, demonstrating the absence of cercariae compatible with Schistosoma mansoni . Nevertheless, the accurate diagnosis of S. mansoni circulation in former endemic areas is still necessary.

Larval trematodes hosted by Biomphalaria straminea in the Brazilian semiarid region: implications for schistosomiasis control

@#The present study aimed to identify larval trematodes shed by snails found in water bodies used by urban communities in a former schistosomiasis endemic area in the state of Piauí, in the Brazilian semiarid region. A malacological survey was performed followed by analysis of the cercariae shed by the snails after light exposure. Biomphalaria straminea specimens (n=1,224) were obtained from all seven collection sites. Cercariae shed by snails were i) single tailed, in which one type of cercariae was identified (Echinostoma cercariae), and ii) with bifurcated tail (brevifurcate apharyngeate distome, brevifurcate pharyngeate distome, and longifurcate pharyngeate distome [strigeocercaria]). Brevifurcate apharyngeate distome were further examined and the presence of spikes in swimming membranes enabled the identification of Spirorchiidae cercariae in all individuals, demonstrating the absence of cercariae compatible with Schistosoma mansoni. Nevertheless, the accurate diagnosis of S. mansoni circulation in former endemic areas is still necessary.

Nanoscale physico-mechanical properties of an aging resistant ZTA composite.

OBJECTIVE: To characterize the effects of aging on the nanomechanical properties and 3D surface topographical parameters of an experimental Zirconia Toughened Alumina (ZTA) composite compared to its respective individual counterpart materials. METHODS: Disk-shaped specimens comprised of three material groups were processed: 1) ZTA 70/30 (70% alumina reinforced with 30% second-generation 3Y-TZP); 2) Zpex (Second-generation 3Y-TZP), and; 3) Al2O3 (High purity Alumina) (n = 10/material, 12 × 1 mm). After synthesis, ceramic powders were pressed, the green-body samples were sintered and polished. Nanoindentation testing was performed to record elastic modulus (E) and hardness (H). Interferometry was utilized to assess 3D surface roughness parameters (Sa, Sq), while X-ray diffraction (XRD) and scanning electron microscope (SEM) assessed the crystalline content and microstructure. All tests were performed before and after simulated aging (134°C, 2.2 bar, 20 h). Statistical analyses were performed using linear mixed-model and least square difference pos-hoc tests (α = 5%). RESULTS: XRD spectra indicated increase of monoclinic peaks for Zpex (~18%) relative to ZTA 70/30 (~2.5%) after aging. Additionally, aging did not affect the surface roughness parameters of ZTA 70/30 and Al2O3, although a significant increase in Sa was recorded for Zpex following aging (~90 nm) (p < 0.001). Al2O3 yielded the highest H and E values (H:21 GPa, E: 254 GPa), followed by ZTA 70/30 (H: 13 GPa, E: 214 GPa) and Zpex (H:11 GPa, E: 167 GPa), all significantly different (p < 0.03). CONCLUSION: ZTA 70/30 and Al2O3 presented high hydrothermal stability with respect to all evaluated variables, where artificial aging significantly increased the monoclinic content and surface roughness of Zpex.

Microstructural, mechanical, and optical characterization of an experimental aging-resistant zirconia-toughened alumina (ZTA) composite.

OBJECTIVE: To evaluate the effect of aging on the microstructural, mechanical, and optical properties of an experimental zirconia-toughened alumina composite with 80%Al2O3 and 20%ZrO2 (ZTA Zpex) compared to a translucent zirconia (Zpex) and Alumina. METHODS: Disc-shaped specimens were obtained by uniaxial and isostatic pressing the synthesized powders (n = 70/material). After sintering and polishing, half of the specimens underwent aging (20 h, 134 °C, 2.2 bar). Crystalline content and microstructure were evaluated using X-ray diffraction and scanning electron microscopy, respectively. Specimens underwent biaxial flexural strength testing to determine the characteristic stress, Weibull modulus, and reliability. Translucency parameter (TP) and Contrast ratio (CR) were calculated to characterize optical properties. RESULTS: ZTA Zpex demonstrated a compact surface with a uniform dispersion of zirconia particles within the alumina matrix, and typical alumina and zirconia crystalline content. ZTA Zpex and alumina exhibited higher CR and lower TP than Zpex. ZTA Zpex and Zpex showed significantly higher characteristic stress relative to alumina. While aging did not affect optical and mechanical properties of ZTA Zpex and alumina, Zpex demonstrated a significant increase in translucency, as well as a in characteristic stress. Alumina reliability was significantly lower than others at 300 MPa, ZTA Zpex and Zpex reliability decreased at 800 MPa, except for aged Zpex. SIGNIFICANCE: While aging did not affect the mechanical nor the optical properties of ZTA Zpex and alumina, it did alter both properties of Zpex. The results encourage further investigations to engineer ZTA as a framework material for long span fixed dental prostheses specially where darkened substrates, such as titanium implant abutments or endodontically treated teeth, demand masking.

Aging resistance of an experimental zirconia-toughened alumina composite for large span dental prostheses: Optical and mechanical characterization.

PURPOSE: To synthesize a zirconia-toughened alumina (ZTA) composite with 85% alumina matrix reinforced by 15% zirconia and to characterize its optical and mechanical properties before and after artificial aging, to be compared with a conventional dental zirconia (3Y-TZP). MATERIAL AND METHODS: After syntheses, ZTA and 3Y-TZP powders were uniaxially and isostatically pressed. Green-body samples were sintered and polished to obtain 80 disc-shaped specimens per group (12 × 1 mm, ISO 6872:2015). The crystalline content and microstructure were characterized by X-ray diffraction (XRD) and scanning electron microscope (SEM). Optical properties were determined by the calculation of contrast ratio (CR) and translucency parameter (TP) using reflectance data. Mechanical properties were assessed by Vickers hardness, fracture toughness and biaxial flexural strength test (BFS). All analyses were conducted before and after artificial aging (20h, 134 °C, 0.22 MPa). Optical parameters and microhardness differences were evaluated through repeated-measures analysis of variance (p < 0.05). BFS data were analyzed using Weibull statistics (95% CI). RESULTS: The synthesis of the experimental ZTA composite was successful, with 98% of theoretical density, as shown in the SEM images. XRD patterns revealed typical zirconia and alumina crystalline phases. ZTA optical properties parameters showed no effect of aging, with a high CR and low TP values denoting a high masking-ability. 3Y-TZP presented lower masking-ability and aging significantly affected its optical properties. ZTA Vickers hardness, fracture toughness and Weibull parameters, including characteristic stress and Weibull modulus were not influenced by aging, while 3Y-TZP presented a significant decrease in characteristic stress and increase in fracture toughness after aging. The ZTA probability of survival for missions of 300 and 500 MPa was estimated at ~99% validating its use for 3-unit posterior fixed dental prostheses (FDP), and no different from conventional 3Y-TZP. At high-stress mission (800 MPa) a significant decrease in probability of survival was observed for aged 3Y-TZP (84%) and for immediate and aged ZTA (73 and 82% respectively). CONCLUSION: The ZTA composite presented a dense microstructure, with preservation of the crystalline content, optical and mechanical properties after artificial aging, which encourages future research to validate its potential use for large span FDP.

IgG1+ B-cell immunity predates IgE responses in epicutaneous sensitization to foods.

BACKGROUND: The generation of IgE-mediated food allergy in humans is silent and only diagnosed upon manifestation of clinical symptoms. While experimental models have been used to investigate some mechanisms of allergic sensitization, the generation of humoral immunity and memory remains to be elucidated. Here, we defined the evolution of allergen-specific B-cell responses during epicutaneous sensitization to foods. METHODS: Wild-type and genetic knockout animals, and drug or antibody strategies for cell depletion and immunoglobulin signaling blockade were used to investigate epicutaneous sensitization and disease progression; we analyzed allergen-specific germinal centers and IgG1+ memory B cells by flow cytometry, evaluated humoral responses, and determined clinical reactivity (anaphylaxis). RESULTS: Epicutaneous sensitization caused microscopic skin damage, inflammation, and recruitment of activated dendritic cells to the draining lymph nodes. This process generated allergen-specific IgG1+ germinal center B cells, serum IgG1, and anaphylaxis that was mediated by the alternative pathway. Whether we used peanut and/or ovalbumin from the egg white for sensitization, the allergen-specific IgG1+ memory compartment predominantly exhibited an immature, pro-germinal center phenotype (PDL-2- CD80- CD35+ CD73+ ). Subsequent subclinical exposures to the allergen induced IgE+ germinal center B cells, serum IgE, and likely activated the classical pathway of anaphylaxis. CONCLUSIONS: Our data demonstrate that IgG1+ B-cell immunity against food allergens in epicutaneous sensitization precedes the generation of IgE responses. Therefore, the assessment of allergen-specific cellular and humoral IgG1+ immunity may help to identify individuals at risk of developing IgE-mediated food allergy and hence provide a window for therapeutic interventions.

Kato-Katz thick smears as a DNA source of soil-transmitted helminths.

Despite the reduction in the prevalence of soil-transmitted helminthiases in many regions of the world, morbidity rates remain high in some rural regions. The Kato-Katz technique is a simple, inexpensive and field-applicable tool commonly used for the diagnosis and worm-burden characterization of these infections. Molecular studies have revolutionized our understanding of the epidemiology and evolutionary genetics of parasites. In this study we recovered helminthic DNA from Kato-Katz slides (n = 93) prepared in 2011 in the Brazilian Amazon. We achieved DNA recovery by polymerase chain reaction (PCR) in 84% of cases for Ascaris sp. and 75% of cases for hookworms. The sequencing confirmed the specific species of the amplicons. The slides stored for a few years could be analysed using this methodology, allowing access to DNA from a large collection of samples. We must consider the Kato-Katz thick smears as a source of helminth DNA. This can significantly reduce logistical difficulties in the field in terms of obtaining, preserving, transporting and initial processing of samples.

Cytotoxicity and antitumoral activity of dichloromethane extract and its fractions from Pothomorphe umbellata.

The cytotoxicity of the dichloromethane crude extract (DCE), obtained from the aerial parts of Pothomorphe umbellata (L.) Miq (Piperaceae), was evaluated against nine human cancer cell lines (MCF-7, NCI-ADR/RES, OVCAR-3, PC-3, HT-29, NCI-H460, 786-O, UACC-62, K-562). The DCE presented antiproliferative activity with good potency against all cell lines at low concentrations (between 4.0 and 9.5 microg/mL) and with selectivity (1.55 microg/mL) for the leukemia cell line (K-652). DCE (100, 200, 300 and 400 mg/kg, ip) was also evaluated in the Ehrlich ascites tumor model. Both the survival number and the life span of the animals that died increased by at least 45 and 50%, respectively (8 animals per group), demonstrating P. umbellata extract potential anticancer activity. The results of the in vivo antitumor activity prompted the fractionation of the crude extract. The crude extract was submitted to dry column chromatography with dichloromethane-methanol (99:1). The column effluent fractions were extracted with methanol, dried under vacuum yielding fractions FR1 (less polar), FR2 (medium polarity), and FR3 (polar), which were analyzed for their growth inhibition or cytotoxic properties by a 48-h sulforhodamine B cell viability assay by measuring the total protein content. FR1 demonstrated high potency and cytotoxicity, a result compatible with the high toxicity of oxalic acid; FR2, containing 4-nerolidylcathecol, presented the lowest cytotoxic activity compared to the other two fractions but with selectivity for prostate cancer cell line; FR3, containing a mixture of steroids described in the literature as possessing various biological activities, also presented potent anticancer in vitro activity. These results suggest that P. umbellata DCE in vivo antitumor activity may be a consequence of the activity of different active principles.

Cytotoxicity and antitumoral activity of dichloromethane extract and its fractions from Pothomorphe umbellata

The cytotoxicity of the dichloromethane crude extract (DCE), obtained from the aerial parts of Pothomorphe umbellata (L.) Miq (Piperaceae), was evaluated against nine human cancer cell lines (MCF-7, NCI-ADR/RES, OVCAR-3, PC-3, HT-29, NCI-H460, 786-O, UACC-62, K-562). The DCE presented antiproliferative activity with good potency against all cell lines at low concentrations (between 4.0 and 9.5 µg/mL) and with selectivity (1.55 µg/mL) for the leukemia cell line (K-652). DCE (100, 200, 300 and 400 mg/kg, ip) was also evaluated in the Ehrlich ascites tumor model. Both the survival number and the life span of the animals that died increased by at least 45 and 50 percent, respectively (8 animals per group), demonstrating P. umbellata extract potential anticancer activity. The results of the in vivo antitumor activity prompted the fractionation of the crude extract. The crude extract was submitted to dry column chromatography with dichloromethane-methanol (99:1). The column effluent fractions were extracted with methanol, dried under vacuum yielding fractions FR1 (less polar), FR2 (medium polarity), and FR3 (polar), which were analyzed for their growth inhibition or cytotoxic properties by a 48-h sulforhodamine B cell viability assay by measuring the total protein content. FR1 demonstrated high potency and cytotoxicity, a result compatible with the high toxicity of oxalic acid; FR2, containing 4-nerolidylcathecol, presented the lowest cytotoxic activity compared to the other two fractions but with selectivity for prostate cancer cell line; FR3, containing a mixture of steroids described in the literature as possessing various biological activities, also presented potent anticancer in vitro activity. These results suggest that P. umbellata DCE in vivo antitumor activity may be a consequence of the activity of different active principles.

A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease.

Several recombinant clones expressing antigens from Echinococcus granulosus were isolated previously from a parasite cDNA library using cystic hydatid disease (CHD) patients' sera or rabbit hyperimmune antiserum against a lipoproteic fraction from bovine cyst fluid. Six of these antigens were expressed in Escherichia coli and the purified recombinant proteins were tested in enzyme-linked immunosorbent assay (ELISA) for specific IgG with a panel of sera from patients with surgically confirmed (n = 58) or immunologically diagnosed (n = 71) CHD. Sera from clinically normal individuals (n = 203) and sera from individuals with other helminthic infections (n = 65) were assayed for the assessment of specificity. A cut-off value was determined by receiver-operating-characteristic plots for each antigen. A recombinant antigen B subunit (AgB8/2) presented the highest sensitivity (93.1%), considering the group of sera from patients with CHD surgically confirmed, and specificity (99.5%) and is proposed as the basis for an immunodiagnostic test. The other recombinant antigens tested presented sensitivities between 58.6% and 89.7%, and three of them were considered of complementary value. In subclass-specific ELISA, different IgG isotypes showed dominance in the response for each of the recombinant antigens. There was a clear predominance of IgG4 response for all antigens tested, indicating that this would be the subclass of choice to be assessed for these recombinant proteins.

Evidence suggesting that gonadal hormones influence benzodiazepine withdrawal-induced weight loss in rats.

1. In experimental animals, benzodiazepine (BZ) withdrawal syndrome includes anorexia and acute weight loss. The literature shows several sex-based differences in the expression of BZ dependence; however, the authors did not find studies dealing with the influence of gonadal hormones on BZ withdrawal-induced weight loss. Thus, this study was designed to investigate the effects of castration on diazepam (DZ) withdrawal-induced weight loss in rats. 2. Male (260-330 g) or female (220-260 g) Wistar rats were anesthetized with ether and submitted to surgical castration or sham-operation. Seven days later, recovered from the surgery, the animals were injected i.p. with DZ (4 mg kg-1 day-1) or appropriate vehicle (VEH; 2 ml kg-1 kg-1 day-1) for 28 days. In the next 7 days, the rats received the same doses of DZ (four groups) or VEH (eight groups). Weights of all animals were recorded daily to the nearest gram at 09:00 h. To assess the degree of weight loss and make statistical comparisons, weights over days 29-34 were expressed as percentage of those recorded in the morning of day 28. 3. Sham-operated female rats from the group DZ-VEH showed a small but statistically significant weight loss on days 29 and 30 (P < 0.05) when compared with groups VEH-VEH and DZ-DZ. Ovariectomized rats, however, did not show any significant change in body weight from days 29-34. 4. Sham-operated male rats did not exhibit any significant weight loss after DZ withdrawal. Orchidectomized animals, however, showed a small but statistically significant weight loss on day 31 (P < 0.05) when compared with groups VEH-VEH and DZ-DZ. 5. These results show a gonadal influence on DZ withdrawal-induced weight loss in Wistar rats; in particular, they also suggest that female hormones (progesterone and/or estrogen) facilitate whilst male hormones inhibit this phenomenon.

Endothelial injury provokes collateral arterial vasoconstriction: response to a serotonin 2 antagonist, thromboxane antagonist or synthetase inhibition.

To assess the influence on collateral arteries of vasoactive factors released from activated platelets we used angiography and blood flow measurement to study the limb blood supply in 29 rabbits, 2 weeks after superficial femoral artery ligation. Minutes after balloon catheter injury to the lower aorta, striking spasm of the collateral arteries was routinely evident on the arteriograms, and limb blood flow fell. Spasm was partly reversed either by blockade of thromboxane synthesis (UK-38,485) or its vascular action (SQ 29,548) or by ketanserin, the serotonin receptor antagonist when used alone. Ketanserin combined with either UK-38,485 or SQ 29,548 reversed the spasm completely. We conclude that a combined action of serotonin and thromboxane induces collateral artery spasm when platelets are activated.

Intravenous digital subtraction angiography in diagnosis of vascular graft occlusion.

Differentiation is often difficult between vascular graft occlusion and progression of underlying disease in patients after vascular surgery. We have studied 57 patients after surgery for traumatic and atherosclerotic arterial occlusion and other vascular anomalies using a commercial digital subtraction angiography (DSA) unit; no complications occurred. Indications for examination included pain, diminished pulse, and failure of catheter angiography. Graft patency was established if proximal and distal anastomoses were visualized; occlusion was diagnosed if no graft was imaged or vascular stump found--noted in 31 grafts. Our diagnosis was proved surgically in 24 patients (two refused operation); three others were confirmed angiographically and one by Doppler ultrasonic examination. In our experience DSA is a safe, specific means of following postoperative grafts and diagnosing their occlusion.

Comparison of intravenous digital subtraction angiography to conventional aortography in patients with abdominal aortic aneurysm.

Evaluation of intravenous digital subtraction angiography (IV DSA) in patients with abdominal aortic aneurysm was performed by obtaining catheter aortograms immediately before DSA studies in ten patients. Diagnostic images were obtained in nine of ten digital subtraction examinations. Although repeat injections were necessary in six DSA and three conventional aortography cases for adequate imaging of both cephalad and caudal extension of the aneurysm, average contrast dose was 53 cc (62 cc in standard catheter studies). Renal artery stenosis was diagnosed by DSA in two of three vessels, multiple renal arteries were demonstrated by both modalities in two cases. Digital subtraction and conventional aortographic findings were proved at surgery. Intravenous DSA was shown to be useful in the preoperative evaluation of patients with abdominal aortic aneurysm.

The effect of iodized oil on the TSH response to TRH in endemic goiter patients.

The TSH and T3 response to synthetic TRH was evaluated in 4 groups of patients: normal controls and goitrous subjects from the urban area of Sao Paulo (urinary iodine excretion: 172.2 +/- 48.3 mug I/g creatinine) and nongoitrous and goitrous subjects from the endemic areas of Sao Bento (urinary iodine excretion: 53.8 +/- 17.1 mug I/g). Plasma T4 and T3 were within our normal range in all groups of patients. The mean plasma TSH was significantly higher (5.2 +/- 3.3 muU/ml) in goitrous subjects living in Sao Bento as compared to normal control groups both in urban or endemic areas, and after TRH these patients had an exaggerated and sustained TSH response with a significantly higher peak level (21.1 +/- 7.9 muU/ml). T3 concentration rose in all subjects following TRH and all patients from the Sao Bento endemic areas had a significantly higher proportionate increase in plasma T3 at 120 min. After an injection of iodized oil basal plasma TSH returned to the normal range in the goitrous subjects from Sao Bento. The mean peak TSH response to TRH was 9.1 +/- 3.8 muU/ml at 3 months after the iodized oil injection, and only at 6 months after the iodized oil TSH response was significantly reduced (peak level: 6.1 +/- 2.4 muU/ml). It is confirmed that plasma TSH levels are increased in endemic goitrous patients but not in normal controls living in the same endemic area and it is suggested that the pituitary threshold for inhibition of secretion of TSH by T4 and T3 has been reset in these goitrous subjects to achieve a persistently higher secretion rate of TSH.