Nicotiana tabacum as an experimental host for the study of plant-Xylella fastidiosa interactions

Xylella fastidiosa causes citrus variegated chlorosis (CVC). Information generated from the X. fastidiosa genome project is being used to study the underlying mechanisms responsible for pathogenicity. However, the lack of an experimental host other than citrus to study plant-X. fastidiosa interaction has been an obstacle to accelerated progress in this area. We present here results of three experiments that demonstrated that tobacco could be an important experimental host for X. fastidiosa. All tobacco plants inoculated with a citrus strain of X. fastidiosa expressed unequivocal symptoms, consisting of orange leaf lesions, approximately 2 months after injection of the pathogen. CVC symptoms were observed in citrus 3 to 6 months after inoculation. The pathogen was readily detected in symptomatic tobacco plants by polymerase chain reaction (PCR) and phase contrast microscopy. In addition, X. fastidiosa was reisolated on agar plates in 4 of 10 plants. Scanning electron microscopy analysis of cross sections of stems and petioles revealed the presence of rod shaped bacteria restricted to the xylem of inoculated plants. The cell size was within the limit typical of X. fastidiosa.

PCR-based detection of Babesia bovis and Babesia bigemina in their natural host Boophilus microplus and cattle

PCR and nested-PCR methods were used to assess the frequency of Babesia bovis and Babesia bigemina infection in Boophilus microplus engorged females and eggs and in cattle reared in an area with endemic babesiosis. Blood and the engorged female ticks were from 27 naturally infested calves and 25 crossbred cows. The frequency of both Babesia species was similar in calves and cows (P > 0.05). Babesia bovis was detected in 23 (85.2%) calves and in 25 (100%) cows and B. bigemina was detected in 25 (92.6%) calves and in 21 (84%) cows. Mixed infections with the both Babesia species were identified in 42 animals, 21 in each age category. Of female ticks engorged on calves, 34.9% were negative and single species infection with B. bigemina (56.2%) was significantly more frequent (P < 0.01) than with B. bovis (4.7%). Most of the females (60.8%) engorged on cows did not show Babesia spp. infection and the frequency of single B. bovis infection (17.6%) was similar (P > 0.05) to the frequency of single B. bigemina infection (15.9%). Mixed Babesia infection was lower (P < 0.01) than single species infection in female ticks engorged either in cows (5.7%) or in calves (4.3%). An egg sample from each female was analysed for the presence of Babesia species. Of the egg samples from female ticks infected with B. bovis, 26 (47.3%) were infected while from those from female ticks infected with B. bigemina 141 (76.6%) were infected (P < 0.01). The results showed that although the frequency of both species of Babesia was similar in calves and cows, the infectivity of B. bigemina was higher to ticks fed on calves while to those ticks fed on cows the infectivity of both Babesia species was similar. © 2004 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Photoelastic comparison of single tooth implant-abutment-bone of platform switching vs conventional implant designs

Objectives: The maintenance and stability of peri-implantar soft tissue seem to be related to the crestal bone around the implant platform and different implant designs connections might affect this phenomenon. The aim of this study was to evaluate by photoelastic analysis the stress distribution in the cervical and apical site of implant-abutment interface of conventional implant joints (external hex, internal hex and cone morse) and compare to the novel platform switching design. Materials and methods: It was fabricated photoelastic models using five different implant-abutment connection, one set of external hex (Alvim Ti, Neodent, Curitiba, Brazil), one set of internal hex (Full Osseotite, Biomet 3i, Florida, USA), one cone morse set (Alvim CM, Neodent, Curitiba, Brazil), and two sets of internal hex plus platform switching concept (Alvim II Plus, Neodent, Curitiba, Brazil) (Certain Prevail, Biomet 3i, Florida, USA). These models were submitted to two compressive loads, axial from 20 kgf (load I) and another (load II), inclined 45° from 10 kgf. During the qualitative analysis, digital pictures were taken from a polariscope, for each load situation. For the quantitative analyses in both situations of load, the medium, minimum and maximum in MPa values of shear strain were determined in the cervical and apical site. The Kruskal-Wallis test was used to compare the results between the different systems and between cervical and apical site were compared using Mann-Whitney U test. Results: The results from qualitative analysis showed less concentration of strain in the cervical area to the internal hex plus platform switching (Certain Prevail), in both situation of load. The same results were get in the quantitative analysis, showing less stress concentrations around the implant Certain Prevail with internal hex plus the novel design (17.9 MPa to load I and 29.5 MPa to load II), however, without statistical significant difference between the systems. Conclusion: The minor stress concentration strongly suggest the use of platform switching design as a manner to prevent bone loss around the implant-abutment platform. Clinical Significance: From the result of this study its possible to make clinical decision for implant system which provides implant components with platform switching characteristics.

Poult intestinal organ culture for propagation of Turkey coronavirus and assessment of host-virus interactions

Infection of young poults with turkey coronavirus (TCoV) produces a syndrome characterized by acute enteritis, diarrhea, anorexia, ruffled feathers, decreased body weight gain and uneven flock growth. The objective of this study was to standardize an intestinal organ culture (IOC) in order to assess host-virus interaction related to apoptosis. For this purpose the Brazilian strain (TCoV/Brazil/2006 with GenBank accession number FJ188401), was used for infection. Infected IOC cells had mitochondrial dysfunction and initial nuclear activation with MTT value of 90.7 (± 2.4) and apoptotic factor 2.21 (± 2.1), considered statistically different from uninfected IOC cells (p > 0.05). The kinetics of TCoV antigens and viral RNA was directly correlated to annexin-V, caspases- 2 and -3, p53, BCl-2 antigens at 24, 72 and 96 h post-infection (p.i.). Morphological and biochemical features of apoptosis, such as in situ nuclear fragmentation (TUNEL and annexin-V) and DNA ladder formation were also detected in infected cells at all assayed p.i. intervals. Moreover, different from other coronaviruses, the expression of both effective caspase-2 and - 3 and p53 antigens were considered lower. However, at all p.i., the BCl-2 antigens were expressed quantitatively and qualitatively as viral antigen measured by immunofluorescence microscopy analysis. Because the diagnosis of TCoV infection is only performed by infecting embryonated poult eggs, the pathological characteri tics related to host-virus interaction remain unclear. This is the first report on apoptosis of TCoV infected IOC, and reveals that it may be useful immunological method to assess virus pathogenesis.

A comparison of performance indexes in DC-DC converters under different stabilizing state-dependent switching laws

This paper deals with the problem of establishing stabilizing state-dependent switching laws in DC-DC converters operating at continuous conduction mode (CCM) and comparing their performance indexes. Firstly, the nature of the problem is defined, that is, the study of switched affine systems, which may not share a common equilibrium point. The concept of stability is, therefore, broadened. Then, the central theorem is proposed, from which a family of switching laws can be derived, namely the minimum law and the hold state law. Some of these are proved to stabilize the basic DC-DC converters and then, their performances are compared to another law, from a previous work, by simulation, where a great reduction in overshoot is obtained. © 2011 IEEE.

Influence of microthreads and platform switching on stress distribution in bone using angled abutments

Purpose: To evaluate the stress distribution in peri-implant bone by simulating the effect of an implant with microthreads and platform switching on angled abutments through tridimensional finite element analysis. The postulated hypothesis was that the presence of microthreads and platform switching would reduce the stress concentration in the cortical bone. Methods: Four mathematical models of a central incisor supported by an implant (5.0mm×13mm) were created in which the type of thread surface in the neck portion (microthreaded or smooth) and the diameter of the angled abutment connection (5.0 and 4.1mm) were varied. These models included the RM (regular platform and microthreads), the RS (regular platform and smooth neck surface), the SM (platform switching and microthreads), and the SS (platform switching and smooth neck). The analysis was performed using ANSYS Workbench 10.0 (Swanson Analysis System). An oblique load (100N) was applied to the palatine surface of the central incisor. The bone/implant interface was considered to be perfectly integrated. Values for the maximum (σmax) and minimum (σmin) principal stress, the equivalent von Mises stress (σvM), and the maximum principal elastic strain (e{open}max) for cortical and trabecular bone were obtained. Results: For the cortical bone, the highest σmax (MPa) were observed for the RM (55.1), the RS (51.0), the SM (49.5), and the SS (44.8) models. The highest σvM (MPa) were found for the RM (45.4), the SM (42.1), the RS (38.7), and the SS models (37). The highest values for σmin were found for the RM, SM, RS and SS models. For the trabecular bone, the highest σmax values (MPa) were observed in the RS model (6.55), followed by the RM (6.37), SS (5.6), and SM (5.2) models. Conclusion: The hypothesis that the presence of microthreads and a switching platform would reduce the stress concentration in the cortical bone was partially rejected, mainly because the microthreads increased the stress concentration in cortical bone. Only platform switching reduced the stress in cortical bone. © 2012 Japan Prosthodontic Society.

Differential mixer with NMOS/PMOS stack at switching stage

This paper proposes a novel differential mixer topology. The traditional stage of switching is replaced by a stack of NMOS and PMOS transistors combined. A design is given of a 900 MHz down-conversion mixer using a 0.35 μm CMOS process. Comparison with conventional mixer shows that the topology leads to a better performance in terms of conversion gain and linearity. ©2012 IEEE.

A DC-DC converter based on the three-state switching cell for high current and voltage step-down applications

This paper presents a pulsewidth modulation dc-dc nonisolated buck converter using the three-state switching cell, constituted by two active switches, two diodes, and two coupled inductors. Only part of the load power is processed by the active switches, reducing the peak current through the switches to half of the load current, as higher power levels can then be achieved by the proposed topology. The volume of reactive elements, i.e., inductors and capacitors, is also decreased since the ripple frequency of the output voltage is twice the switching frequency. Due to the intrinsic characteristics of the topology, total losses are distributed among all semiconductors. Another advantage of this converter is the reduced region for discontinuous conduction mode when compared to the conventional buck converter or, in other words, the operation range in continuous conduction mode is increased, as demonstrated by the static gain plot. The theoretical approach is detailed through qualitative and quantitative analyses by the application of the three-state switching cell to the buck converter operating in nonoverlapping mode $(D < 0.5)$. Besides, the mathematical analysis and development of an experimental prototype rated at 1 kW are carried out. The main experimental results are presented and adequately discussed to clearly identify its claimed advantages. © 1986-2012 IEEE.

On Hepatitis C Virus Evolution: The Interaction between Virus and Host towards Treatment Outcome

Background:Hepatitis C is a disease spread throughout the world. Hepatitis C virus (HCV), the etiological agent of this disease, is a single-stranded positive RNA virus. Its genome encodes a single precursor protein that yields ten proteins after processing. NS5A, one of the non-structural viral proteins, is most associated with interferon-based therapy response, the approved treatment for hepatitis C in Brazil. HCV has a high mutation rate and therefore high variability, which may be important for evading the immune system and response to therapy. The aim of this study was to analyze the evolution of NS5A quasispecies before, during, and after treatment in patients infected with HCV genotype 3a who presented different therapy responses.Methods:Viral RNA was extracted, cDNA was synthesized, the NS5A region was amplified and cloned, and 15 clones from each time-point were sequenced. The sequences were analyzed for evolutionary history, genetic diversity and selection.Results:This analysis shows that the viral population that persists after treatment for most non-responder patients is present in before-treatment samples, suggesting it is adapted to evade treatment. In contrast, the population found in before treatment samples from most end-of-treatment responder patients either are selected out or appears in low frequency after relapse, therefore changing the population structure. The exceptions illustrate the uniqueness of the evolutionary process, and therefore the treatment resistance process, in each patient.Conclusion:Although evolutionary behavior throughout treatment showed that each patient presented different population dynamics unrelated to therapy outcome, it seems that the viral population from non-responders that resists the treatment already had strains that could evade therapy before it started. © 2013 Bittar et al.

Infection patterns of Paradollfusnema amphisbaenia (Nematoda: Cosmocercidae) in a population of Amphisbaena wuchereri (Squamata: Amphisbaenidae) from Minas Gerais state, south-eastern Brazil, and its relations with host size, sex and fat body mass

Specimens (n= 41) of the amphisbaenid Amphisbaena wuchereri taken from a population in Minas Gerais state, south-eastern Brazil, were examined for gastrointestinal parasites. A single nematode species was found, Paradollfusnema amphisbaenia. This was a new host record for this nematode species. This parasite was encountered in the large intestine (prevalence of 100%), in the stomach (prevalence of 2%) and in the small intestine (prevalence of 7.3%). The intensity of infection ranged from 1 to 457 individual parasites per host and was positively correlated with body size of both male and female amphisbaenians. The discrepancy index (D) indicated that P. amphisbaenia tended to an even distribution in this host population. The nematode, which did not affect fat body mass, induced inflammatory infiltrations in the small intestine, indicating that the parasites might injure the host's organs. Copyright © Cambridge University Press 2012.

Regular and switching platform: Bone stress analysis with varying implant diameter

The aim of this study was to evaluate stress distribution of the peri-implant bone by simulating the biomechanical influence of implants with different diameters of regular or platform switched connections by means of 3-dimensional finite element analysis. Five mathematical models of an implant-supported central incisor were created by varying the diameter (5.5 and 4.5 mm, internal hexagon) and abutment platform (regular and platform switched). For the cortical bone, the highest stress values (rmax and rvm) were observed in situation R1, followed by situations S1, R2, S3, and S2. For the trabecular bone, the highest stress values (rmax) were observed in situation S3, followed by situations R1, S1, R2, and S2. The influence of platform switching was more evident for cortical bone than for trabecular bone and was mainly seen in large platform diameter reduction.

Virus-host interaction in feline immunodeficiency virus (FIV) infection

Feline immunodeficiency virus (FIV) infection has been the focus of several studies because this virus exhibits genetic and pathogenic characteristics that are similar to those of the human immunodeficiency virus (HIV). FIV causes acquired immunodeficiency syndrome (AIDS) in cats, nevertheless, a large fraction of infected cats remain asymptomatic throughout life despite of persistent chronic infection. This slow disease progression may be due to the presence of factors that are involved in the natural resistance to infection and the immune response that is mounted by the animals, as well as due to the adaptation of the virus to the host. Therefore, the study of virus-host interaction is essential to the understanding of the different patterns of disease course and the virus persistence in the host, and to help with the development of effective vaccines and perhaps the cure of FIV and HIV infections. © 2013 Elsevier Ltd. All rights reserved.

The platform switching concept revisited. An experimental study in dogs

Objective: To evaluate the influence of the configuration of the marginal aspect of implants placed immediately into extraction sockets on peri-implant hard tissue adaptation. Material and methods: In 6 Labrador dogs, endodontic treatments of the mesial roots of 1M1 were performed and the distal roots were removed. 2P2 was extracted as well. Implants were immediately placed in the center of the distal alveoli. Cylindrical straight implants were installed in the right side of the mandible (Control), while, in the left side, implants with a reduced diameter in the coronal portion, yielding an indentation in the surface continuity (Test), were installed. Cover screws were affixed, and the flaps were sutured to allow non-submerged healing. After 4 months of healing, histological slides were obtained for assessments. Results: A buccal resorption of 1.58 ± 1.28 and 1.90 ± 1.93 mm at the control and of 0.26 ± 0.90 and 0.14 ± 0.66 mm at the test sites was observed at the premolar and molar regions, respectively. The buccal coronal level of osseointegration was located apically to the margin of the smooth/rough surface border by 2.40 ± 0.90 and 3.70 ± 0.87 mm at the control sites and 1.19 ± 0.45 and 2.16 ± 0.96 mm at the test sites at the premolar and molar sites, respectively. All differences yielded statistical significance. Conclusions: The use of implants with a reduced diameter in their coronal aspect may contribute to preservation of the buccal bony crest in a more coronal level compared with conventional implants. Thus, the study confirmed the efficacy of the platform switching concept. © 2013 John Wiley & Sons A/S.

Lack of host specificity of Colletotrichum spp. isolates associated with anthracnose symptoms on mango in Brazil

The aim of the present study was to analyse the genetic and pathogenic variability of Colletotrichum spp. isolates from various organs and cultivars of mango with anthracnose symptoms, collected from different municipalities of São Paulo State, Brazil. Colletotrichum gloeosporioides isolates from symptomless citrus leaves and C. acutatum isolates from citrus flowers with post-bloom fruit drop symptoms were included as controls. Sequencing of the ITS region allowed the identification of 183 C. gloeosporioides isolates from mango; only one isolate was identified as C. acutatum. amova analysis of ITS sequences showed larger genetic variability among isolates from the same municipality than among those from different populations. fAFLP markers indicated high levels of genetic variability among the C. gloeosporioides isolates from mango and no correlation between genetic variability and isolate source. Only one C. gloeosporioides mango isolate had the same genotype as the C. gloeosporioides isolates from citrus leaves, as determined by ITS sequencing and fAFLP analysis. Pathogenicity tests revealed that C. gloeosporioides and C. acutatum isolates from either mango or citrus can cause anthracnose symptoms on leaves of mango cvs Palmer and Tommy Atkins and blossom blight symptoms in citrus flowers. These outcomes indicate a lack of host specificity of the Colletotrichum species and suggest the possibility of host migration. © 2012 British Society for Plant Pathology.