Genetic diversity of Brazilian isolates of feline immunodeficiency virus

FAPESP (the Sao Paulo State research funding foundation)

Life cycle of Ixodes luciae (Acari: Ixodidae) in the laboratory

The life cycle of Ixodes luciae was evaluated for five consecutive generations in the laboratory. Wild mice Calomys callosus and laboratory rats Rattus norvegicus were used as hosts for larvae and nymphs. For adult ticks, opossums Didelphis aurita were used as hosts. Off-host developmental periods were observed in an incubator at 27A degrees C and 95% RH. The life cycle of I. luciae lasted 95-97 days, excluding prefeeding periods. C. callosus, one of the natural host species for I. luciae immature stages, was shown to be much more suitable than the artificial host R. norvegicus. Significantly (P < 0.05), more larvae and nymphs successfully fed on C. callosus than on R. norvegicus. When tick-na < ve C. callosus were exposed to three consecutive larval infestations at 24-day intervals, recovery of engorged larvae were greater in the second and third infestations, indicating that previous infestations did not induce acquired resistance to ticks. Larval feeding period typically varied from 5 to 10 days on R. norvegicus, but was significantly (P < 0.05), longer on C. callosus (range, 7-34 days). The majority (71.7%) of I. luciae adult females successfully fed and oviposited after exposed to D. aurita. Mean engorged weight (581.9 mg; range, 237.1-796.0 mg) of these females were much higher than those previously reported for other New World Ixodes species. Our results are in accordance to the current literature that appoints opossums Didelphidae and small rodents (e.g., C. callosus) natural hosts for I. luciae immature and adult stages, respectively.

Biology and life cycle of Amblyomma incisum (Acari: Ixodidae)

Amblyomma incisum Neumann is a major tick species in the Atlantic Forest of Brazil. Tapir is the main host for adult ticks and a high aggressiveness of nymphs to humans has been reported. In this work data on the biology and life cycle of this tick species is presented for the first time. It was shown that horse is a suitable host for A. incisum adults and rabbit for larvae and nymphs. It was also shown that A. incisum is a big tick species (mean engorged female weight of 1.96 g) with a long life cycle which lasts 262.3 days when maintained at 27A degrees C and 85% RH. These laboratory conditions were, however, inappropriate and egg hatching rate (1.2%) was very low. Nevertheless egg hatching of ticks in a forest patch increased considerably (72.2%) indicating that this A. incisum population is highly dependent on a forest-like environment.

Redescription of the female, description of the male, and several new records of Amblyomma parkeri (Acari: Ixodidae), a South American tick species

The tick Amblyomma parkeri Fonseca and Arago was described in 1952, based on female and immature ticks collected in the states of So Paulo and Santa Catarina, Brazil. Thereafter, there has been no further report of A. parkeri, and the male has remained unknown. Herein, we examined ticks collected on porcupines from a locality in the state of So Paulo. Some of the ticks were identified as Amblyomma longirostre (Koch, 1844), whereas others as A. parkeri, including male specimens, for which we provide the first description. We also provide additional reports of A. parkeri after examining collections of A. longirostre and Amblyomma geayi Neumann, 1899 from different tick collections. Morphological evidence to support the original description of A. parkeri is presented, supported by molecular analyses of portions of the 16S rRNA and 12S rRNA mitochondrial genes. Morphological particularities to separate A. parkeri, A. longirostre, and A. geayi are provided.

Haplotypes of the bovine IgG2 heavy gamma chain in tick-resistant and tick-susceptible breeds of cattle

Bovines present contrasting, heritable phenotypes of infestations with the cattle tick, Rhipicephalus (Boophilus) microplus. Tick salivary glands produce IgG-binding proteins (IGBPs) as a mechanism for escaping from host antibodies that these ectoparasites ingest during blood meals. Allotypes that occur in the constant region of IgG may differ in their capacity to bind with tick IGBPs; this may be reflected by the distribution of distinct allotypes according to phenotypes of tick infestations. In order to test this hypothesis, we investigated the frequency of haplotypes of bovine IgG2 among tick-resistant and tick-susceptible breeds of bovines. Sequencing of the gene coding for the heavy chain of IgG2 from 114 tick-resistant (Bos taurus indicus, Nelore breed) and tick-susceptible (B. t. taurus, Holstein breed) bovines revealed SNPs that generated 13 different haplotypes, of which 11 were novel and 5 were exclusive of Holstein and 3 of Nelore breeds. Alignment and modeling of coded haplotypes for hinge regions of the bovine IgG2 showed that they differ in the distribution of polar and hydrophobic amino acids and in shape according to the distribution of these amino acids. We also found that there was an association between genotypes of the constant region of the IgG2 heavy chain with phenotypes of tick infestations. These findings open the possibility of investigating if certain IgG allotypes hinder the function of tick IGBPs. If so, they may be markers for breeding for resistance against tick infestations.

In vitro evaluation of erbium, chromium:yttrium-scandium-gallium-garnet laser-treated enamel demineralization

This study evaluated the effect of different parameters of erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation on enamel mineral loss in a simulated caries model. Forty-five enamel samples obtained from third molar teeth (3 mmx 3 mm) were randomly divided into five groups (n = 9): G1-Er,Cr:YSGG laser at 0.25 W, 20 Hz, 2.8 J/cm(2); G2-Er,Cr:YSGG laser at 0.50 W, 20 Hz, 5.7 J/cm(2); G3-Er,Cr:YSGG laser at 0.75 W, 20 Hz, 8.5 J/cm(2); G4-sodium fluoride (NaF) dentifrice (positive control); G5-no treatment (negative control). After irradiation, the samples were submitted to 2 weeks of pH cycling. After the acid challenge, the samples were assessed by cross-sectional microhardness at different depths from the enamel surface. Analysis of variance (ANOVA) and Student-Newman-Keuls tests were performed (alpha = 5%). The percentage of lesion inhibition for each group was: G1 37%; G2 38%; G3 64%, and G4 50.5%. Regarding the relative mineral loss values (micrometers x volume percent), groups G1 (1,392 +/- 522) and G2 (1,292 +/- 657) did not differ significantly from each other, but both had higher values than group G3 (753 +/- 287); the groups irradiated with Er,Cr:YSGG laser did not differ from group G4. Although the findings of the study revealed that Er,Cr:YSGG laser irradiation at 8.5 J/cm(2) can be an alternative for the enhancement of the enamel`s resistance to acid, lower energy densities also produced a cariostatic potential comparable to the use of fluoride dentifrice.

Laser phototherapy effect on protein metabolism parameters of rat salivary glands

The aim of this study was to evaluate the effects of infrared diode laser phototherapy (LP) on tissues of the submandibular gland (SMG) and parotid gland (PG). Wistar rats were randomly divided into experimental (A and B) and control (C) groups. A diode laser, 808 nm wavelength, in continuous wave mode, was applied to the PG, SMG and sublingual gland in the experimental groups on two consecutive days. The doses were 4 J/cm(2) and 8 J/cm(2), and total energy was 7 J and 14 J, respectively. The power output (500 mW) and power density (277 mW/cm(2)) were the same for both experimental groups. In order to visualize the area irradiated by the infrared laser, we used a red pilot beam (650 nm) with 3 mW maximum power for the experimental groups. For the control group, the red pilot beam was the only device used. The SMG and PG were removed after 1 week of the first irradiation. Total protein concentration, amylase, peroxidase, catalase and lactate dehydrogenase assays were performed, as well as histological analysis. Statistical tests revealed significant increase in the total protein concentration for groups A and B in the parotid glands (P < 0.05). Based on the results of this study, LP altered the total protein concentration in rats` parotid glands.

The neuroprotective effect of dental pulp cells in models of Alzheimer`s and Parkinson`s disease

Aim of the present study was to investigate the neuroprotective effect of dental pulp cells (DPCs) in in vitro models of Alzheimer and Parkinson disease. Primary cultures of hippocampal and ventral mesencephalic neurons were treated for 24 h with amyloid beta (A beta(1-42)) peptide 1-42 and 6-OHDA, respectively. DPCs isolated from adult rat incisors were previously cultured in tissue culture inserts and added to the neuron cultures 2 days prior to neurotoxin treatment. Cell viability was assessed by the MTT assay. The co-culture with DPCs significantly attenuated 6-OHDA and A beta(1-42)-induced toxicity in primary cultures of mesencephalic and hippocampal neurons, and lead to an increase in neuronal viability in untreated cultures, suggesting a neurotrophic effect in both models. Furthermore, human dental pulp cells expressed a neuronal phenotype and produced the neurotrophic factors NGF, GDNF, BDNF, and BMP2 shown by microarray screening and antibody staining for the representative proteins. DPCs protected primary neurons in in vitro models of Alzheimer`s and Parkinson`s disease and can be viewed as possible candidates for studies on cell-based therapy.

Erbium, chromium : yttrium scandium gallium garnet laser for caries removal: influence on bonding of a self-etching adhesive system

This study evaluated the influence of the dental substrates obtained after the use of different caries removal techniques on bonding of a self-etching system. Forty, extracted, carious, human molars were ground to expose flat surfaces containing caries-infected dentine surrounded by sound dentine. The caries lesions of the specimens were removed or not (control-G1) either by round steel burs and water-cooled, low speed, handpiece (G2), or by irradiation with an erbium, chromium:yttrium scandium gallium garnet (Er,Cr:YSGG) laser (2W, 20 Hz, 35.38 J/cm(2), fiber G4 handpiece with 0.2826 mm(2), non-contact mode at a 2 mm distance, 70% air/20% water-G3) or using a chemo-mechanical method (Carisolv-G4). Caries-infected, caries-affected and sound dentines were submitted to a bonding system followed by construction of a resin-based composite crown. Hour-glass shaped samples were obtained and submitted to a micro-tensile bond test. The bond strength data were compared by analysis of variance (ANOVA), complemented by Tukey`s test (P <= 0.05). The samples of sound dentine presented higher bond strengths than did samples of caries-affected dentine, except for the groups treated with the Er,Cr:YSGG laser. The highest bond strengths were observed with the sound dentine treated with burs and Carisolv. The bond strengths to caries-affected dentine were similar in all groups. Additionally, bonding to caries-affected dentine of the Er,Cr:YSGG laser and Carisolv groups was similar to bonding to caries-infected dentine. Thus, caries-affected dentine is not an adequate substrate for adhesion. Moreover, amongst the caries removal methods tested, the Er,Cr:YSGG laser irradiation was the poorest in providing a substrate for bonding with the tested self-etching system.

Low-fluence CO(2) laser irradiation decreases enamel solubility

This study investigated whether subablative-pulsed CO(2) laser (10.6 mu m) irradiation, using fluences lower than 1 J/cm(2), was capable of reducing enamel acid solubility. Fifty-one samples of bovine dental enamel were divided into three groups: control group, which was not irradiated (CG); group laser A (LA) irradiated with 0.3 J/cm ; and group laser B (LB) irradiated with 0.7 J/cm(2). After irradiation, the samples were subjected to demineralization in an acetate buffer solution and were then analyzed by SEM. A finite-element model was used to calculate the temperature increase. The calcium and phosphorous content in the demineralization solution were measured with an ICP-OES. ANOVA and the t-test pairwise comparison (p < 0.016) revealed that LB showed significantly lower mean Ca and P content values in the demineralization solution than other groups. A reduction in the enamel solubility can be obtained with pulsed CO(2) laser irradiation (0.7 J/cm(2), 135 mJ/pulse, 74 Hz, 100 mu s) without any surface photomodification and a less than 2 degrees C temperature increase at a 3-mm depth from the surface.

Effect of defocused infrared diode laser on salivary flow rate and some salivary parameters of rats

This study aims to investigate whether infrared diode low-level laser therapy (LLLT) increased salivary flow rate and altered pH value, protein concentration, and peroxidase and amylase activities in saliva of rats. Wistar rats were used and divided into three groups. Experimental groups (A and 13) had their parotid, submandibular and sublingual glands submitted to diode laser, 808-nm wavelength, on two consecutive days. The dose results were 4 and 8 J/cm(2), respectively. A red guide light was used to visualize the irradiated area. Group C was irradiated only with red pilot beam and served as control. The saliva samples were collected after each irradiation step (first and second collection days) and 1 week after the first irradiation (seventh day). Statistical analysis was performed, and differences were observed according to different days of salivary collection. The results showed that salivary flow rate for groups A and B was higher on the seventh day if it is compared to data obtained for the first day (p<0.05). LLLT applications on salivary glands are a therapy procedure that requires further studies.

Comparison of dentin root canal permeability and morphology after irradiation with Nd:YAG, Er:YAG, and diode lasers

The aim of this study was to compare the effects of Nd:YAG, Er:YAG, and diode lasers on the morphology and permeability of root canal walls. The three laser wavelengths mentioned interact differently with dentin and therefore it is possible that the permeability changes caused will determine different indications during endodontic treatment. Twenty-eight human single-rooted teeth were instrumented up to ISO 40 and divided into four groups: group C, control (GC), non-laser irradiated; group N (GN), irradiated with Nd:YAG laser; group E (GE), with Er:YAG laser and group D (GD) with diode laser. After that, the roots were filled with a 2% methylene blue dye, divided into two halves and then photographed. The images were analyzed using Image J software and the percentage of dye penetration in the cervical, middle, and apical root thirds were calculated. Additional scanning electron microscopy (SEM) analyses were also performed. The analysis of variance (ANOVA) showed significant permeability differences between all groups in the middle and cervical thirds (p < 0.05). The Tukey test showed that in the cervical third, GN presented means of dye penetration statistically significantly lower than all of the other groups. In the middle third, GE and GD showed statistically higher dye penetration means than GC and GN. SEM analysis showed melted surfaces for GN, clean wall surfaces with open dentinal tubules for GE, and mostly obliterated dentinal tubules for GD. Er:YAG (2,094 nm) laser and diode laser (808 nm) root canal irradiation increase dentinal permeability and Nd:YAG (1,064 nm) laser decreases dentin permeability, within the studied parameters.

Shrinkage stress and mechanical properties of photoactivated composite resin using the argon ion laser

The objective of this study was to verify the influence of photoactivation with the argon ion laser on shrinkage stress (SS), followed by evaluation of Vickers microhardness (VM), percentage of maximum hardness (PMH), flexural strength (FS), and flexural modulus (FM) of a composite resin. The study groups were: L1-laser at 200 mW for 10 seconds; L2-laser at 200 mW for 20 seconds; L3-laser at 250 mW for 10 seconds; L4-laser at 250 mW for 20 seconds; H-halogen light at 275 mW for 20 seconds. Data were analyzed by ANOVA/Tukey`s test (alpha=5%). The values of SS (MPa) were statistically lower for the group L3 (1.3)c, followed by groups L1 (2.7)b, L4 (3.4)a, b, L2 (3.7)a, and H (4.5)a. There was no difference in the values of VM when the same time of photoactivation was used, with respective values being L1=70.1a, L2=78.1b, L3=69.9a, L4=78.1b and H=79.9b. All groups showed a PMH of at least 80%. Only the group L1 showed differences in FS (MPa) and FM (GPa), the respective values of 86.2 and 5.4 being lower. Therefore, the use of argon ion laser had influenced the composite resin polymerization. The L3 group presented adequate mechanical properties and minimum SS, reducing the clinical working time for photoactivation of restorations with the tested resin by 50%.