Establishment and cryptic transmission of Zika virus in Brazil and the Americas.

Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.

Short communication: Does previous superovulation affect fertility in dairy heifers?

The aim of this study was to evaluate the potential negative effects of superovulation on subsequent fertility of dairy heifers. Holstein heifers (n = 1,783), 312 to 387 d of age, and 273 to 307 kg of body weight (BW) from 2 commercial farms were enrolled. These animals were first selected to be donors (446) or nondonors (CON, n = 1,327) according to their genomic values. Nondonor heifers (CON) were artificially inseminated (AI) according to standard procedures of each farm after reaching 320 kg of BW. Donor heifers were superovulated using a fixed FSH dose (180 mg NIH-FSH-P1; Folltropin-V, Vetoquinol Brasil, Mairiporã, SP, Brazil) and embryos were collected following standard procedures. Heifers that produced fewer than 8 viable embryos after first superovulation (SOV1, n = 337) were no longer used as donors, whereas the remaining heifers (SOV2, n = 109) were superovulated a second time within an interval of 48 to 54 d. Donor heifers (SOV1 and SOV2) were AI once they reached 320 kg of BW, at least 15 d after the last embryo collection. Data on age at first AI, at conception, and at parturition, as well as the number of services per conception, were analyzed by ANOVA, using the PROC MIXED procedure of SAS (SAS Institute Inc., Cary, NC) procedure. Binomial variables (pregnancy per AI, overall pregnancy rate, open heifers at 500 d age, and late pregnancy loss) were analyzed using the GLIMMIX procedure of SAS. The heifers selected to undergo superovulation twice (SOV2) yielded more total (12.6 ± 5.3 vs. 6.8 ± 4.6; respectively) and viable embryos (8.5 ± 3.8 vs. 3.9 ± 2.8; respectively) than those superovulated only once (SOV1). Age at first AI, conception, and at parturition was greater in SOV2, but not in SOV1 compared with nondonor controls. In addition, pregnancy per first AI, overall pregnancy rate, services per conception, open heifers at 500 d of age, and occurrence of pregnancy losses after 60 d of gestation were similar among CON, SOV1, and SOV2 heifers. In summary, a single superovulation performed before heifers reach a minimum weight for breeding did not affect age at conception, calving or other indicators of reproductive efficiency. On the other hand, heifers superovulated twice were first inseminated at a later age than their birth cohorts, but had similar reproductive performance.

Freezing and thawing in Antarctica: characterization of antifreeze protein (AFP) producing microorganisms isolated from King George Island, Antarctica.

Antarctic temperature variations and long periods of freezing shaped the evolution of microorganisms with unique survival mechanisms. These resilient organisms exhibit several adaptations for life in extreme cold. In such ecosystems, microorganisms endure the absence of liquid water and exhibit resistance to freezing by producing water-binding molecules such as antifreeze proteins (AFP). AFPs modify the ice structure, lower the freezing point, and inhibit recrystallization. The objective of this study was to select and identify microorganisms isolated from different Antarctic ecosystems based on their resistance to temperatures below 0 °C. Furthermore, the study sought to characterize these microorganisms regarding their potential antifreeze adaptive mechanisms. Samples of soil, moss, permafrost, and marine sediment were collected on King George Island, located in the South Shetland archipelago, Antarctica. Bacteria and yeasts were isolated and subjected to freezing-resistance and ice recrystallization inhibition (IR) tests. A total of 215 microorganisms were isolated, out of which 118 were molecularly identified through molecular analysis using the 16S rRNA and ITS regions. Furthermore, our study identified 24 freezing-resistant isolates, including two yeasts and 22 bacteria. A total of 131 protein extracts were subjected to the IR test, revealing 14 isolates positive for AFP production. Finally, four isolates showed both freeze-resistance and IR activity (Arthrobacter sp. BGS04, Pseudomonas sp. BGS05, Cryobacterium sp. P64, and Acinetobacter sp. M1_25C). This study emphasizes the diversity of Antarctic microorganisms with the ability to tolerate freezing conditions. These microorganisms warrant further investigation to conduct a comprehensive analysis of their antifreeze capabilities, with the goal of exploring their potential for future biotechnological applications.

Color Doppler flow imaging for the early detection of nonpregnant cattle at 20 days after timed artificial insemination.

The objective was to determine the accuracy of a pregnancy test for predicting nonpregnant cattle based on the evaluation of corpus luteum (CL) blood flow at 20 d (CLBF-d20) after timed artificial insemination (TAI). Crossbred Holstein-Gir dairy heifers (n=209) and lactating cows (n=317) were synchronized for TAI using the following protocol: intravaginal implant (1.0 g of progesterone) and 2mg of estradiol benzoate i.m. on d -10, implant removal and 0.526 mg of sodium cloprostenol i.m. on d -2, 1mg of estradiol benzoate i.m. on d -1, and TAI on d 0. On d 20, animals underwent grayscale ultrasonography (US) to locate the CL and color flow Doppler to evaluate CLBF-d20 using a portable ultrasound equipped with a 7.5-MHz rectal transducer. Based only on a visual, subjective CLBF evaluation, the animals were classified as pregnant or not pregnant. On d 30 to 35, blinded from results of the previous diagnosis, the same operator performed a final pregnancy diagnosis using US to visualize the fetal heartbeat (gold standard; US-d30). A second evaluator also analyzed the CLBF-d20 in the same animals by watching 7-s recorded videos. Blood samples were collected from a subset of 171 females to determine, by RIA, plasma progesterone (P4) concentrations, which indicate CL function. The final pregnancy outcome (US-d30) was retrospectively compared with the CLBF-d20 diagnoses and then classified either as correct or incorrect. The number of true positive, true negative, false positive, and false negative decisions were inserted into a 2 × 2 decision matrix. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the CLBF-d20 test were calculated using specific equations. Binomial variables (pregnancy rate and proportions) were analyzed using Fisher's exact test for the effect of parity and to compare between evaluators and tests (CLBF-d20 vs. plasma P4). The kappa values were calculated to quantify the agreement between CLBF-d20 and the gold standard (US-d30) and between evaluators. The performance parameters of CLBF-d20 test were as follows: sensitivity=99.0%, specificity=53.7%, positive predictive value=65.1%, negative predictive value=98.5%, and accuracy=74.8%. False negatives represented only 0.4% of the exams. No differences existed in these parameters between evaluators (no. 1 vs. no. 2) and tests (CLBF-d20 vs. plasma P4). Moreover, a high level of agreement was observed between evaluators (0.91). In conclusion, visual evaluation of CLBF-d20 represents a quick, reliable, and consistent diagnostic test that enables the early detection of nonpregnant cattle.

Blind Source Separation in Persistent Atrial Fibrillation Electrocardiograms Using Block-Term Tensor Decomposition With Löwner Constraints.

The estimation of the atrial activity (AA) signal from electrocardiogram (ECG) recordings is an important step in the noninvasive analysis of atrial fibrillation (AF), the most common sustained arrhythmia encountered in clinical practice. This problem admits a blind source separation (BSS) formulation that has been recently posed as a tensor factorization, using the Hankel-based block term decomposition (BTD), which is particularly well-suited to the estimation of exponential models like AA during AF. However, persistent forms of AF are characterized by short R-R intervals and very disorganized (or weak) AA, making it difficult to model AA directly and perform its successful extraction through Hankel-BTD. To overcome this drawback, the present work proposes a tensor approach to estimate QRS complexes and subtract them from the ECG, resulting in a signal that, ideally, only contains the AA component. Such an approach tackles the problem of blind separation of rational functions, which models QRS complexes explicitly. The data tensor admitting a BTD is built from Löwner matrices generated from each lead of the observed ECG. To this end, this paper formulates a variant of the recently proposed constrained alternating group lasso (CAGL) algorithm that imposes Löwner structure on the decomposition blocks. This is done by performing an orthogonal projection, which we explicitly derive, at each iteration of CAGL. Results from experiments with synthetic data show the consistency of the proposed Löwner-constrained AGL (LCAGL) in extracting the desired sources. Experimental results obtained on a population of 20 patients suffering from persistent AF show that the proposed variant outperforms other tensor-based methods in terms of atrial signal estimation quality from ECG records as short as a single heartbeat.

Early resynchronization of non-pregnant beef cows based in corpus luteum blood flow evaluation 21 days after Timed-AI.

The study aimed to verify whether a hormone protocol started at Day 13 (D13) after Timed Artificial Insemination (TAI) influences the conception rate. Nelore cows (primiparous and multiparous) from two commercial beef farms (n = 1,431) were first TAI (D0). Timed AI was performed in lots (TAI Lots) ranging from 187 to 346 cows. On D13, regarding the TAI lot, cows were assigned for either receiving (Resynch group, n = 1,002) or not (Control group, a subset of approximately 30%, n = 429) another hormone protocol for resynchronization. The same hormone protocol was used for the first TAI and for the resynchronization, except for 1 mg instead of 2 mg of estradiol benzoate (EB) at the begging of the protocol. Eight days later (D21), the Resynch group was checked for corpus luteum blood flow by color Doppler ultrasonography, and in those detected as non-pregnant, the protocol was completed and a 2nd TAI was performed at D23. Pregnancy diagnosis was later (D30) performed by B-mode ultrasonography in the control group and confirmed in the presumptive pregnant cows from the 1st TAI of the Resynch group. The remaining cows were checked for pregnancy 30 days after the 2nd TAI (experimental Day 53). The statistical model to explain conception rate considered the effects of Group (Control or Resynch), Farm, Parity (primiparous or multiparous), Sire, Technician (who perform AI), TAI Lot and pertinent interactions (Group*Parity, Group*Farm and Group*TAI Lot). The statistical analyses of the model were performed using the Proc Glimmix (SAS virtual University Edition). The conception rate for the 1st TAI was similar (P > 0.4) between Control (50.3%, 216/429) and Resynch group (52.6%, 527/1002). The positive predictive diagnostic on D21 showed high relation with PD30 (90.7%, 527/581). In Resynch group, non-pregnant cows (n = 421, 1002 minus 581) were re-inseminated. The conception rate of the 2nd TAI (42.8%, 180/421) was affected (P < 0.002) by side effects of the Farm (48.5 vs. 33.1%) and Parity (51.2 vs. 40.3%, for multiparous vs. primiparous, p < 0.001). Nevertheless, after the 2 TAIs of the Resynch group, the cumulative conception rate was 70.5% (707/1002). In conclusion, the early resynchronization of cows with a low (1 mg) EB dose and progesterone device at D13 after TAI can be used as a strategy to reduce conception interval in beef cattle, and thus to increase the number of pregnant cows from artificial insemination after the breeding season.

Crystallization and preliminary X-ray diffraction analysis of crotoxin B from Crotalus durissus collilineatus venom.

Crotoxin B is a basic phospholipase A(2) found in the venom of several Crotalus durissus ssp. rattlesnakes and is one of the subunits that constitute crotoxin, the main component of the venom of these snakes. This heterodimeric toxin is related to important envenomation effects such as neurological disorders, myotoxicity and renal failure. Although crotoxin was first crystallized in 1938, the first structural data only became available in 2007 (for crotoxin B from C. durissus terrificus) and showed an ambiguous result for the biological assembly, which could be either dimeric or tetrameric. In this work, the crystallization, X-ray diffraction data collection at 2.2 A resolution and molecular-replacement solution of a dimeric complex formed by two crotoxin B isoforms from C. durissus collilineatus venom is presented.

Differential expression of LHCGR and its isoforms is associated to the variability in superovulation responses of Gir cattle.

The aim of this study was to evaluate the pattern of expression of LHCGR isoforms in Gir heifers characterized as good (10.3 ±â€¯1.2 ova/embryos per flush, n = 5) or poor responders (1.1 ±â€¯0.3 ova/embryos per flush, n = 5) to superovulation protocols. In both groups, an adapted ultrasound-guided follicular aspiration system was used to collect granulosa cells from 8 mm follicles formed either during a synchronized, non-stimulated follicular wave (no stimulation control, NS) or on the fourth day of a superovulation protocol (SOV) induced with 200 IU of pFSH. The recovered follicular fluid was centrifuged and granulosa cells were washed with NaCl 0.9% and kept in RNAlater®. RNA extraction was performed using a commercial RNeasy Micro Kit and eluted samples were quantified and reverse transcribed using the commercial Superscript III kit. cDNA samples were amplified by real-time PCR using a primer to target LH/hCG receptor gene - not selective for LHCGR isoforms (total LHCGR) - and four sets of isoforms selective primers (S1, S10, S10 + 11, and S11). Analyses were performed using the REST software and expression levels are shown as mean ± SEM. Under physiological conditions (NS), poor responders had a higher expression of total LHCGR (4.9 ± 1.7 fold-change, P < 0.01) as well as isoforms S10, S11 and S10 + 11, compared to good responders. In both phenotypes, superovulation down-regulated total LHCGR expression (-0.5 ± 0.2 and -0.9 ± 0.0 for good and poor responders, respectively; P < 0.05). However, in poor responders the exogenous FSH treatment up-regulated the S10 (2.4 ± 2.0; P < 0.05), S10 + 11 (3.8 ± 3.2; P < 0.01), and S1 isoforms (1.8 ± 1.3; P < 0.05), compared to good responders We conclude that down-regulation of total LHCGR, associated to up-regulation of their inactive isoforms, may have compromised follicle development and thus contributed to the low efficiency of superovulation in heifers with a poor responder phenotype.

Crystallization and preliminary X-ray diffraction analysis of a myotoxic Lys49-PLA2 from Bothrops jararacussu venom complexed with p-bromophenacyl bromide.

For the first time, a non-catalytic and myotoxic Lys49-PLA2 (BthTX-I from Bothrops jararacussu venom) has been crystallized with BPB inhibitor. X-ray diffraction data were collected and electron-density calculations showed that the ligand is bound to the His48 residue. BthTX-I with His48 chemically modified by BPB shows strongly reduced myotoxic and cytotoxic activities. This suggests a biological correlation between the modification of His48, which is associated with catalytic activity of PLA2s, and other toxicological activities of Lys49-PLA2s.

Oocyte development and fecundity type of the Brazilian Snapper Lutjanus alexandrei Moura & Lindeman, 2007 (Perciformes: Lutjanidae).

Lutjanid species exhibit multiple spawning behaviour during an extended spawning season in warm months, asynchronous oocyte development and indeterminate fecundity. Although early studies have contributed to knowledge of the reproductive cycle of many species within the group, they have not considered aspects about the number of cortical alveoli oocyte stage throughout maturity phases along spawning season. The latter aspect is also considered very important to confirm indeterminate fecundity hypothesis. In the present study, were analyzed 154 Brazilian snapper Lutjanus alexandrei female gonads obtained from artisanal fisheries in Pernambuco State (Brazil) between October 2010 and March 2011. Were measured oocyte size frequency distribution for maturity phases (developing, spawning capable and actively spawning), and oocyte development stage (unyolked oocytes, cortical alveoli, primary, secondary and tertiary vitellogenic oocytes and hydrated oocytes), and also the oocyte stage frequency during spawning season. The frequency of cortical alveoli oocyte stage was constantly found in the spawning period (>37%), showing slight variation throughout maturity phases. The absence of gap in the oocyte size frequency distribution between primary and secondary oocyte growth stages during spawning season is a strong indicator of continuous oocyte recruitment from reserve stocks. In addition, co-occurrence of tertiary vitellogenic oocytes, hydrated oocytes, post-ovulatory follicles and yellow-brown bodies in the histological sections of ovaries reinforce indeterminate fecundity hypothesis.

Efficacy of induction of luteolysis in superovulated cows is dependent on time of prostaglandin F2alpha analog treatment: effects on plasma progesterone and luteinizing hormone profiles.

The objectives were to (1) evaluate the effectiveness of induction of luteolysis in superovulated (SOV) cows at two distinct time points after embryo flushing; and (2) compare the pattern of LH release after treatment with PGF in cows with single vs. multiple ovulations. In the first experiment, Holstein cows were SOV with 400 IU of FSH following standard procedures. Uterine flushing for embryo recovery was performed 7 days after artificial insemination (Day 0), and cows were randomly allocated into two groups to receive PGF (0.5-mg sodium cloprostenol, intramascular) either immediately after flushing (Day 7 group, N = 19) or 4 days later (Day 11 group, N = 20). Time of luteolysis was determined on the basis of plasma progesterone (P4) concentrations. There was no difference (P > 0.05) in plasma P4 before treatment between Day 7 and Day 11 groups. A decline in plasma P4 was observed 48 hours after PGF treatment in both the groups (P < 0.0001). In Day 11 cows, P4 continued to decrease thereafter, whereas Day 7 animals had no further reduction in plasma P4. Luteolysis (P4 < 1 ng/mL) occurred in all Day 11 cows. In the Day 7 group, however, luteolysis failure was observed for 11 of 19 cows (57.9%). In cows without luteolysis, plasma P4 increased after the initial PGF-induced decline. The second experiment compared luteolysis in (SOV, N = 6) vs. non-SOV (control, N = 8) cows. Both groups received a single PGF treatment on Day 11 after estrus, and luteolysis was monitored daily by ovarian ultrasonography and plasma P4 measurements. In addition, plasma LH was measured in blood samples taken every 20 minutes for 1 hour during five consecutive days after treatment. A similar percentage of reduction in P4 was observed in both groups 24 hours after treatment; however, SOV cows only reached plasma P4 values similar (P > 0.05) to controls 96 hours after treatment. There was no difference in initial LH values between SOV and controls (P > 0.05). The slower decrease in plasma P4 in the SOV group prevented an increase in LH for up to 96 hours after luteolysis induction, whereas LH values increased (P < 0.05) in controls 24 hours after treatment. In conclusion, (1) luteolysis may fail or be incomplete when PGF treatment is given on the day of uterine flushing (Day 7) in SOV cows; (2) induction of luteolysis 4 days later (Day 11) is effective, but the initial high-plasma P4 concentrations result in a slower slope of P4 decline to basal levels, and consequently, delayed increase in LH pulses.

Corpus luteum blood flow evaluation on Day 21 to improve the management of embryo recipient herds.

The aim of the present study was to use blood flow evaluation of the CL at 14 days after embryo transfer to detect nonpregnant animals and optimize the management of bovine recipients. The estrous cycle was synchronized in 165 recipients, and the day of expected ovulation was considered to be Day 0. Embryo transfer was performed 7 days later, on Day 7. On Day 21, pregnancy was diagnosed on the basis of blood flow evaluation of the CL (DG21-predictive diagnostic). To validate this methodology, visual scores for blood flow were compared to objective data extracted from CL ultrasound images recorded in the Doppler mode. The size was also evaluated using recorded images of the CL in the B mode. Blood samples were also collected for further analysis of the progesterone (P4) concentration. The diagnosis of pregnancy was confirmed at 35 days after estrus (DG35-definitive diagnostic). The DG21 showed that 55.2% (90 of 163) of the animals were presumptively pregnant, and this value was higher (P < 0.04) than that obtained at DG35 (43.6%, 71 of 163). The predictive diagnostic achieved moderate specificity (79.3%) for the detection of pregnancy, but most importantly, high sensitivity (100%) for the detection of nonpregnant recipients. The overall accuracy of the diagnosis was 88.3%. The P4 concentrations were different (P < 0.02) and correlated with each visual score assigned for the CL size. Visual scores for CL blood flow were also efficient (P < 0.0001) to distinguish animals with different levels of P4; however, P4 concentrations were higher for scores 1 and 2 (high and regular blood flow, respectively) than those for score 3 (low blood flow). This technique showed high sensitivity and facilitated the early detection of nonpregnant animals. The DG21 would allow about 79.3% of nonpregnant animals to be resynchronized 9 to 14 days earlier, when compared to conventional management based on pregnancy diagnosis at Days 30 to 35.

Effectiveness of Different Mechanical Methods on Dentin Caries Removal: Micro-CT and Digital Image Evaluation.

PURPOSE: To determine the caries removal effectiveness (CRE) and minimal invasive potential (MIP) of caries excavation methods using digital imaging and microtomography analyses. METHODS: Twelve human molars with occlusal caries lesions in dentin were randomly divided into three groups (carbide bur, exacavator, and polymer bur). They were sectioned mesiodistally, and standardized digital and computed microtomography x-ray (micro-CT) images were taken from each section before and after caries excavation. On each image, initial carious dentin (IC), prepared cavity (PC), and residual caries (RC) were defined according to visual criteria using ImageJ software. CRE was determined based on the RC/IC ratio, whereas MIP was determined by the PC/IC ratio. Data were analyzed using one-way analysis of variance and Student t-test or with Kruskal-Wallis and Student-Newman-Keuls test. The level of significance was set at 0.05. RESULTS: For both digital image and micro-CT analysis, the carbide bur showed higher CRE values than the excavator (p=0.0063 and p=0.0263, respectively) and the polymer bur (p=0.0028 and p=0.0005, respectively). The latter two presented similar results (p>0.05). Regarding MIP, for the digital image analysis, the polymer bur was different from the carbide bur (p=0.0030) but was not different from that of the excavator (p=0.1240). For micro-CT analysis, the MIP values of all the groups were significantly different, and the polymer bur was the most conservative method (p<0.05). CONCLUSIONS: The carbide bur was the most effective method for caries removal but was not completely conservative. The polymer bur and excavator presented low invasive potential but were not able to remove all of the carious dentin.

Effects of prism orientation on tensile strength of enamel.

PURPOSE: This study tested the hypothesis that the tensile strength of enamel varies according to prismatic orientation. MATERIALS AND METHODS: Eight extracted, caries-free human third molars were thoroughly cleaned with pumice and water. The entire enamel surface was conditioned with 37% phosphoric acid for 30 s, air dried, and bonded with Single Bond adhesive system. Several layers of composite (Z-100) were incrementally applied to the crown to build up a "cube-like" resin structure approximately 5 mm thick covering the entire crown of the teeth. The teeth were stored for 24 h in water at 37 degrees C, and then the crown was serially, vertically sectioned in a mesio-distal direction to obtain several slices approximately 0.7 mm thick. The slices were sectioned into halves, and each half was gently trimmed from both sides with a diamond bur to reduce the cross-sectional area to a "neck" located in enamel, either on the external slope or functional slope of the cusps. The specimens were trimmed to permit testing of enamel with its prisms either oriented parallel (PL) or perpendicular (PD) to the applied load. The trimmed specimens were glued to the fixtures of a Vitrodyne tester and stressed in tension at 0.5 mm/min. SEM images were taken from the fractured surfaces to examine the site of failure and confirm the prism orientation. RESULTS: Mean tensile strength of enamel was 24.7 +/- 9.6 MPa (n = 22) for PL and 11.4 +/- 6.3 MPa (n = 22) for PD oriented enamel prisms (t value = -5.45, p < 0.05). There was no significant difference between specimens originating from different slopes of the cusps (p > 0.05). CONCLUSION: The results showed that tensile strength of enamel is dependent on the prismatic orientation.

Tensile strength of human dentin as a function of tubule orientation and density.

PURPOSE: This study investigated the ultimate tensile strength (UTS) of human dentin as a function of tubule orientation and density. MATERIALS AND METHODS: Slabs of dentin (ca 0.7 mm thick) were obtained from human third molars by either transversally or longitudinally sectioning the crowns with an Isomet saw. The slabs were gently trimmed to reduce the central area of the coronal dentin to a cross-sectional area of approximately 0.5 mm2. The longitudinally sectioned slabs were either trimmed from the mesial and distal sides or from the occlusal and pulpal aspects to permit the tensile load to be applied either parallel or perpendicular to the tubule orientation. The transversally sectioned specimens were obtained at several distances from the pulp and were used to evaluate the effects of tubule density. The trimmed specimens were tested in tension on a Kratos testing machine at 0.5 mm/min. After failure, the UTS of each was calculated and expressed in MPa. The fractured ends of the transversally sectioned specimens were viewed under SEM to calculate the number of tubules per mm2 at the site of fracture, and its relation with the UTS was investigated by regression analysis. RESULTS: The UTS of dentin is higher when the load is applied perpendicular to the tubule orientation (80 +/- 13 MPa) than when applied parallel to tubule orientation (58 +/- 11 MPa, p < 0.05). There was a tendency for dentin to be weaker as the number of tubules at the site of fracture increased, although this relationship was not statistically significant (R2 = 0.051, p > 0.05). CONCLUSION: The UTS of dentin is dependent on the tubule direction. Dentin tends to be weaker as the number of tubules per area increases.

The microtensile bond test: a review.

PURPOSE: The purpose of this review is to describe all of the various modifications of the microtensile bond test in one paper, so that investigators can select the modification that best suits their needs. METHODS: The essence of the microtensile test is the division of resin-bonded teeth into slabs between 0.5 and 1.0 mm thick that are then trimmed in such a manner that tensile force will be concentrated on the bonded interface during testing. Among the many advantages of the technique are that each tooth produces multiple specimens. Further, there is no need for a matrix to limit the bonded surface area, since the area is determined by the dimensions of the trimmed specimens. RESULTS: The various modifications of the microtensile test have been used to measure differences in regional bond strength across occlusal dentin, down the external surface of teeth from crown through roots, down the internal surface of root canals from cervical to apical thirds, as well as to compare normal vs caries-affected occlusal dentin and normal vs sclerotic cervical dentin. The technique is ideal for evaluating the long-term durability of resin-hard-tissue bonds. CONCLUSION: The microtensile test methods offer versatility that cannot be achieved by conventional methods. It is more labor-intensive than conventional testing, but holds great potential for providing insight into the strength of adhesion of restorative materials to clinically relevant sites and substrates.

Investigation of complexity of the instruction manuals for electrical coffeepots.

BACKGROUND: Electrical coffeepots are commonly used in professional and residential environments. Their instruction manuals are related to issues that involve the user's safety and ability to operate the machine correctly. To provide the best product performance to the user, one must indicate or inform the correct usability, to turn the interaction easier. OBJECTIVE: This research proposes to investigate the instruction manuals of the electrical coffeepots. METHODS: Four coffee pot instruction manuals were analyzed in relation to the complexity through the heuristic evaluation. For that, eight experts of the Graphic Design were chosen to answer twenty four questions with the aim of analyzing: images; texts; layout development; information and warnings. RESULTS: This study shows the results of the 04 (four) items analyzed: a) images; b) texts; c) layout development; d) information and warnings, together with the suggestions of improvements for each manual. CONCLUSIONS: It is believed that the methodological procedures for the application of the heuristic evaluation have facilitated the diagnosis of fragilities and barriers that the users find during the interaction with electrical coffeepot manuals.

Efficiency of the spermatogenesis in zebu bulls (Bos taurus indicus).

The objective of the research was to evaluate the efficiency of the spermatogenesis through the morphology of the testicular parenchyma in bulls of different zebu breeds. We used testicular fragments from bull of the breeds Nelore (n = 10), Polled Nelore (n = 6), Gyr (n = 5), Guzerat (n = 5) and Tabapuã (n = 5). The tissue was perfused with Karnovsky solution, included in glycol methacrylate and stained with toluidine blue-sodium borate 1%. Animals of the Nelore breed presented higher population of primary spermatocyte in pre-leptotene/leptotene (38.30) and in pachytene (38.14) and round spermatids (113.30), higher yield of spermatogonia mitosis (21.2) and higher daily spermatic production per gram of testicular parenchyma (32.8 × 10(6) ) than those from breeds Gyr, Guzerat and Tabapuã and higher general yield of spermatogenesis (62.4) than breeds Gyr and Tabapuã. There was no significant difference in any of the evaluated parameters between breeds Nelore and Polled Nelore. The rate of Sertoli cells did not vary between the studied breeds. Apparently, the genetic selection applied to the breeds has been improving the yield in the spermatogenic process by decreasing cellular loss, although it did not increase the support capacity of the Sertoli cells.

Efficacy and limitations of different approaches to anticipate the diagnosis of pregnancy in cattle / Eficácia e limitações de diferentes abordagens para antecipar o diagnóstico de gestação em bovinos

The study evaluated sonographic and serologic exams performed for early (20 to 30d) diagnosis of pregnancy. One hundred-twenty (n= 120) bovine recipients were synchronized (estrous=D0) and timed embryo transferred (TET, D7) with fresh in vitro produced embryos. In the first trial (n= 46), diagnosis of pregnancy was performed on day 20 (D20) by detecting CL blood flow (BF) and by Pregnancy-Associated Glycoproteins (PAGs) serology. In the second trial (n= 30), pregnancy diagnosis was performed on D25 by ultrasound visualization of uterine contents and by PAGs serology. In the last trial, PAG's serology was performed on D30. Ultrasonographic detection of the uterine contents and embryo viability performed on D30 (DG30) was considered the gold standard. The PROC FREQ procedure was used to test the agreement between diagnostic methods. On D20, the Doppler ultrasonography of the CL had showed high sensitivity (100%), but only moderate specificity (53.3%). On the same day, serologic diagnostic had no agreement (k= -0.08, P< 0.46) with the gold standard, with very low sensitivity (6.3%). However, the sensitivity of the serologic exam increased dramatically (from 6.3 to 100%) from D20 to D25, and it contributed to detect false negatives from the ultrasound diagnosis, improving the overall accuracy from 90% to 96.7%.(AU)

O estudo foi planejado para correlacionar exames ultrassonográficos e sorológicos realizados para o diagnóstico precoce (20 a 30d) de gestação. Cento e vinte (n= 120) receptoras bovinas foram sincronizadas (estro=D0), e embriões frescos produzidos in vitro foram transferidos em tempo fixo (TETF, D7). No experimento 1 (n= 46), o diagnóstico de gestação foi realizado no D20, pela detecção do fluxo sanguíneo do CL e pela sorologia de glicoproteínas associadas à gestação (PAGs). No experimento 2 (n= 30), a detecção da gestação foi realizada por meio da visualização do conteúdo do útero e também pela sorologia para PAGs. No experimento 3, a sorologia para PAGs foi realizada no D30. Em todos os experimentos, a visualização ultrassonográfica da vesícula e da viabilidade embrionária, realizada no D30, foi considerada padrão-ouro. O procedimento PROC FREQ testou o nível de concordância dos métodos diagnósticos. No D20, o diagnóstico baseado na vascularização do CL mostrou alta sensibilidade (100%) e apenas moderada especificidade (53,3%). Nesse mesmo dia, o diagnóstico sorológico não apresentou concordância (k=-0,08, P<0,46) com o padrão-ouro, além de baixa sensibilidade (6,3%). No entanto, a sensibilidade do exame sorológico aumentou drasticamente (6,3 para 100%) do D20 para o D25, contribuindo para detectar falsos negativos diagnosticados pela ultrassonografia, melhorando a acurácia (90 para 96,7%).(AU)

Alkylation of histidine residues of Bothrops jararacussu venom proteins and isolated phospholipases A2: a biotechnological tool to improve the production of antibodies.

Crude venom of Bothrops jararacussu and isolated phospholipases A2 (PLA2) of this toxin (BthTX-I and BthTX-II) were chemically modified (alkylation) by p-bromophenacyl bromide (BPB) in order to study antibody production capacity in function of the structure-function relationship of these substances (crude venom and PLA2 native and alkylated). BthTX-II showed enzymatic activity, while BthTX-I did not. Alkylation reduced BthTX-II activity by 50% while this process abolished the catalytic and myotoxic activities of BthTX-I, while reducing its edema-inducing activity by about 50%. Antibody production against the native and alkylated forms of BthTX-I and -II and the cross-reactivity of antibodies to native and alkylated toxins did not show any apparent differences and these observations were reinforced by surface plasmon resonance (SPR) data. Histopathological analysis of mouse gastrocnemius muscle sections after injection of PBS, BthTX-I, BthTX-II, or both myotoxins previously incubated with neutralizing antibody showed inhibition of the toxin-induced myotoxicity. These results reveal that the chemical modification of the phospholipases A2 (PLA2) diminished their toxicity but did not alter their antigenicity. This observation indicates that the modified PLA2 may provide a biotechnological tool to attenuate the toxicity of the crude venom, by improving the production of antibodies and decreasing the local toxic effects of this poisonous substance in animals used to produce antivenom.