Glial reactivity in dogs with visceral leishmaniasis: correlation with T lymphocyte infiltration and with cerebrospinal fluid anti-Leishmania antibody titres

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of TNF-alpha, IL-4, and IL-10 and parasite density in spleen and liver of L. (L.) chagasi naturally infected dogs

Dogs are the main domestic reservoirs of L. (L.) chagasi. Once in the vertebrate host, the parasite can cause visceral leishmaniasis, which can also be transmitted to humans. Cytokines are key elements of the host immune response against Leishmania spp. To investigate whether tumor necrosis factor (TNF)-alpha, interleukin (IL)-4 and IL-10 are associated with pattern infection in dogs, these cytokines were quantified in the spleen and liver of dogs naturally infected with L. (L.) chagasi, with or without clinical manifestations, and their levels were correlated with the parasite load verified in these organs. A total of 40 adult dogs naturally infected with L. (L.) chagasi were assessed, together with 12 uninfected control dogs. Samples from spleen and liver were used to determine the cytokine levels by capture ELISA and for quantifying parasite load by real-time PCR. Statistical analysis was performed using the minimum Chi square method and group means were compared using the Tukey test. TNF-alpha, IL-4 and IL-10 levels in infected dogs were higher than in control groups; the liver was the main cytokine-producing organ during infection. The level of splenic TNF-alpha showed correlation with parasite load and may represent an important marker for infection process evolution, with the participation of IL-10. These results may contribute to a clearer understanding of the immune response in dogs infected with L. (L.) chagasi, which may lead to the development of prophylactic or preventive measures for these animals.

Diminished CD4+/CD25+T cell and increased IFN-gamma levels occur in dogs vaccinated with Leishmune (R) in an endemic area for visceral leishmaniasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

High levels of serum matrix metalloproteinases in dogs with natural visceral leishmaniosis: A preliminary report

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparative evaluation of enzyme-linked immunosorbent assay based on crude and purified antigen in the diagnosis of canine visceral leishmaniasis in symptomatic and oligosymptomatic dogs

This study evaluated the performance of crude total antigen (CTA) and fucose-mannose ligand antigen (FML) in an enzyme-linked immunosorbent assay for diagnosis of canine visceral leishmaniasis (CVL). The assays used sera from known negative controls (n = 30), clinically symptomatic (n = 30) and oligosymptomatic (n = 30) parasitologically proven infection (by microscopy). Aspirates of popliteal lymph node from infected canines were colleted to score parasitism and compared with the ELISA results. The study indicated that FML used in ELISA provided high sensitivity for detecting oligosymptomatic dogs (90%) and CTA showed greater sensitivity than FML for symptomatic canines (90%). In oligosymptomatic dogs, specificity was 100% for CTA-ELISA, but in symptomatic dogs, FML specificity was higher (96.7%) than CTA-ELISA (93.3%). A significant correlation was observed between the degree of parasitism and the results obtained in CTA-ELISA. Since no available antigen offers 100% specificity and sensitivity for CVL diagnosis, the choice of antigen used must depend on the aim of the investigation. (C) 2008 Elsevier B.V. All rights reserved.

Hemodynamic effects of butorphanol in desflurane-anesthetized dogs

Objective To evaluate the effects of butorphanol on cardiopulmonary parameters in dogs anesthetized with desflurane and breathing spontaneously.Study design Prospective, randomized experimental trial.Animals Twenty dogs weighing 12 +/- 3 kg.Methods Animals were distributed into two groups: a control group (CG) and butorphanol group (BG). Propofol was used for induction and anesthesia was maintained with desflurane (10%). Forty minutes after induction, the dogs in the CG received sodium chloride 0.9% (0.05 mL kg(-1) IM), and dogs in the BG received butorphanol (0.4 mg kg(-1) IM). The first measurements of body temperature (BT), heart rate (HR), arterial pressures (AP), cardiac output (CO), cardiac index (CI), central venous pressure (CVP), stroke volume index (SVI), pulmonary arterial occlusion pressure (PAOP), mean pulmonary arterial pressure (mPAP), left ventricular stroke work (LVSW), systemic (SVR) and pulmonary (PVR) vascular resistances, respiratory rate (fR), and arterial oxygen (PaO(2)) and carbon dioxide (PaCO(2)) partial pressures were taken immediately before the administration of butorphanol or sodium chloride solution (T0) and then at 15-minute intervals (T15-T75).Results In the BG, HR, AP, mPAP and SVR decreased significantly from T15 to T75 compared to baseline. fR was lower at T30 than at T0 in the BG. AP and fR were significantly lower than in the CG from T15 to T75. PVR was lower in the BG than in the CG at T30, while PaCO(2) was higher compared with T0 from T30 to T75 in the BG and significantly higher than in the CG at T30 to T75.Conclusions and clinical relevance At the studied dose, butorphanol caused hypotension and decreased ventilation during desflurane anesthesia in dogs. The hypotension (from 86 +/- 10 to 64 +/- 10 mmHg) is clinically relevant, despite the maintenance of cardiac index.

Serological diagnosis of visceral leishmaniasis by an enzyme immunoassay using protein A in naturally infected dogs

Um ensaio de imunoadsorção enzimática para detecção de anticorpos contra Leishmania chagasi, utilizando antígeno total de formas promastigotas lisados foi desenvolvido. Cinqüenta cães com sintomas clínicos de leishmaniose visceral foram examinados. Esta técnica utilizou anti-IgG de cão conjugado a peroxidase ou proteína A conjugado a peroxidase. Foi verificado que nos animais positivos diagnosticados por exame parasitológico direto o ensaio ELISA utilizando proteína A conjugada a peroxidase (média da densidade óptica ± desvio padrão 2,078 ± 0,631) detecta mais anticorpos do que o sistema utilizando anti-IgG de cão conjugado a peroxidase (média da densidade óptica ± desvio padrão 1,008 ± 0,437), enquanto para os animais negativos o resultado obtido nos dois sistemas de detecção são similares. Esse resultado sugere que o sistema de ELISA utilizando proteína A conjugado a peroxidase pode ser útil na detecção de animais na fase aguda da infecção e desta forma auxiliar na identificação dos animais positivos e no controle desta importante zoonose.

Periodontal tissue response to coverage of root cavities restored with resin materials: A histomorphometric study in dogs

Background: the purpose of this study was to histomorphometrically evaluate the response of periodontal tissues covering Class V resin restorations in dogs.Methods: After raising a mucoperiosteal flap, bony defects measuring 5 x 5 mm were created on the buccal aspect of the canines of five dogs followed by cavity preparations on the root surface measuring 3 x 3 x 1 mm. Before repositioning the flap to cover the bone defect, the cavities were restored with composite resin (CR) or resin-modified glass ionomer cement (RMGIC) or were left unrestored as control (C). The dogs were euthanized 90 days after surgery. Specimens comprising the tooth and periodontal tissues were removed, processed routinely, cut into longitudinal serial sections in the bucco-lingual direction, and stained with hematoxylin and eosin (H&E) or Masson's trichrome. The most central sections were selected for histomorphometric analysis.Results: Histomorphometric analysis revealed apical migration of epithelial tissue onto the restorative materials (RMGIC and CR). The C group presented significantly longer connective tissue attachment (P < 0.05) than the RMGIC and CR groups and significantly higher bone regeneration (P < 0.05) compared to the RMGIC group. Histologically, the cervical third (CT) of all groups had the most marked chronic inflammatory infiltrate.Conclusions: Within the limits of this study, it can be concluded that the restorative materials used exhibit biocompatibility; however, both materials interfered with the development of new bone and the connective tissue attachment process.

Influence of microgap location and configuration on the periimplant bone morphology in submerged implants. An experimental study in dogs

Objectives: The vertical location of the implant-abutment connection influences the periimplant bone morphology. It is unknown, however, whether different microgap configurations cause different bone reactions. Therefore, in this study the bone morphologies of two different implant systems were compared.Material and methods: Three months after tooth extraction in eight mongrel dogs, two grit-blasted screw implants with internal Morse taper connection (ANK group) were placed on one side whereas the contralateral side received two oxidized screw implants with external hex (TIU group). One implant on each side was placed level with the bone (equicrestal), the second implant was inserted 1.5mm below bone level (subcrestal). After 3 months the implants were uncovered. Three months after stage two surgery, histometrical evaluations were performed in order to assess the periimplant bone levels (PBL), the first bone-to-implant contact points (BICP), the width (HBD) and the steepness (SLO) of the bone defect.Results: All implants osseointegrated clinically and histologically. Bone overgrowth of the microgap was seen in ANK implants only. No significant differences between ANK and TIU could be detected in neither vertical position for PBL and BICP. However, a tendency in favor of ANK was visible when the implants were placed subcrestally. In the parameters HBD (ANK equicrestal -0.23mm; TIU equicrestal -0.51mm; ANK subcrestal +0.19mm; TIU subcrestal -0.57mm) and SLO (ANK equicrestal 35.36 degrees; TIU equicrestal 63.22 degrees; ANK subcrestal 20.40 degrees; TIU subcrestal 44.43 degrees) more pronounced and significant differences were noted.Conclusions: Within the limits of this study, it is concluded that different microgap designs cause different shapes and sizes of the periimplant ('dish-shaped') bone defect in submerged implants both in equicrestal and subcrestal positions.

Tooth replantation after use of euro-coffins solution or bovine milk as storage medium: a histomorphometric analysis in dogs

Purpose: Euro-Collins solution was developed for the preservation of organs for transplantation, whose characteristics have raised interest for its use as a storage medium for avulsed teeth before replantation. This study evaluated histologically and morphometrically the healing process of dog teeth replanted after storage in Euro-Collins solution or bovine milk.Materials and Methods: Eighty roots of 4 young adult mongrel clogs were randomly assigned to 4 groups (n = 20) and the root canals were instrumented and obturated with gutta-percha and a calcium hydroxide-based sealer. After 2 weeks, the teeth were extracted and subjected to the following protocols: GI (negative control), replantation immediately after extraction; GII (positive control), bench-drying for 2 hours before replantation; GIII and GIV, immersion in 10 mL of whole bovine milk and Euro-Collins solution at 4 C, respectively, for 8 hours before replantation. The animals were sacrificed 90 days postoperatively. The pieces containing the replanted teeth were subjected to routine processing for histologic and histometric analyses under light microscopy and polarized light microscopy.Results: Root resorption was observed in all groups. GII exhibited the greatest loss of dental structure (P < .01), and inflammatory resorption was predominant in this group. Storage in milk showed poorer results than immediate replantation and storage in Euro-Collins solution (P < .01). The teeth stored in Euro-Collins solution presented similar extension of root resorption and periodontal ligament reorganization to those of immediately replanted teeth.Conclusions: The findings of this study suggest that the Euro-Collins solution is an adequate storage medium for keeping avulsed teeth for up to 8 hours before replantation.

Effect of calcium hydroxide-based materials on periapical tissue healing and orthodontic root resorption of endodontically treated teeth in dogs

This study evaluated periapical tissue healing and orthodontic root resorption of endodontically treated teeth sealed with calcium hydroxide in dogs. The sample consisted of three contralateral pairs of maxillary incisors and two contralateral pairs of mandibular incisors in each of two dogs using a split mouth design. After biomechanical preparation of the teeth in the first group (n = 10), a Ca(OH)(2) dressing was placed for 14 days before root canal filling with Ca(OH)(2)-based sealer (Sealapex) and gutta-percha points. In the second group (n = 10), root canals were obturated immediately after the mechanical preparation with gutta-percha points and zinc oxide and eugenol (ZOE)-based sealer (Endofill). After completion of endodontic treatment, the teeth were moved with an orthodontic appliance with a calibrated force of 200 g, reactivated every 21 days. After 105 days, the animals were killed and the teeth were removed upon completion of active treatment, without a period of recovery, and prepared for histomorphological analysis. All sections of each tooth were graded subjectively on a scale from one to four to obtain the average of the 16 histomorphological parameters analysed. Evaluation of the differences between the two treatment protocols was made with Mann-Whitney U-test. It was observed that the teeth treated with Ca(OH)(2)-based materials provided better outcomes (P = 5%), with complete repair of all root resorption areas, high rate of biological closure of the main canal and apical accessory canals by newly formed cementum, less intense and extensive chronic inflammatory infiltrate, and better organization of the periodontal ligament. Under the tested conditions, Ca(OH)(2)-based materials had a favourable action on periapical tissue healing and repair of orthodontic root resorption in endodontically treated dogs' teeth.

Influence of Microgap Location and Configuration on Radiographic Bone Loss Around Submerged Implants: An Experimental Study in Dogs

Purpose: The vertical location of the implant-abutment connection influences the subsequent reaction of the peri-implant bone. It is not known, however, whether any additional influence is exerted by different microgap configurations. Therefore, the radiographic bone reactions of two different implant systems were monitored for 6 months. Materials and Methods: In eight mongrel dogs, two implants with an internal Morse-taper connection (INT group) were placed on one side of the mandible; the contralateral side received two implants with an external-hex connection (EXT group). on each side, one implant was aligned at the bone level (equicrestal) and the second implant was placed 1.5 mm subcrestal. Healing abutments were placed 3 months after submerged healing, and the implants were maintained for another 3 months without prosthetic loading. At implant placement and after 1, 2, 3, 4, 5, and 6 months, standardized radiographs were obtained, and peri-implant bone levels were measured with regard to microgap location and evaluated statistically. Results: All implants osseointegrated clinically and radiographically. The overall mean bone loss was 0.68 +/- 0.59 mm in the equicrestal INT group, 1.32 +/- 0.49 mm in the equicrestal EXT group, 0.76 +/- 0.49 mm in the subcrestal INT group, and 1.88 +/- 0.81 mm in the subcrestal EXT group. The differences between the INT and EXT groups were statistically significant (paired t tests). The first significant differences between the internal and external groups were seen at month 1 in the subcrestal groups and at 3 months in the equicrestal groups. Bone loss was most pronounced in the subcrestal EXT group. Conclusions: Within the limits of this study, different microgap configurations can cause different amounts of bone loss, even before prosthetic loading. Subcrestal placement of a butt-joint microgap design may lead to more pronounced radiographic bone loss. INT J ORAL MAXILLOFAC IMPLANTS 2011;26:941-946

Influence of Microgap Location and Configuration on Radiographic Bone Loss in Nonsubmerged Implants: An Experimental Study in Dogs

Purpose: The implant-abutment connection (microgap) influences the pen-implant bone morphology. However, it is unclear if different microgap configurations additionally modify bone reactions. This preliminary study aimed to radiographically monitor pen-implant bone levels in two different microgap configurations during 3 months of nonsubmerged healing. Materials and Methods: Six dogs received two implants with internal Morse taper connection (INT group) on one side of the mandible and two implants with external-hex connection (EXT group) on the other side. One implant on each side was positioned at bone level (equicrestal); the second implant was inserted 1.5 mm below the bone crest (subcrestal). Healing abutments were attached directly after implant insertion, and the implants were maintained for 3 months without prosthetic loading. At implant placement and 1, 2, and 3 months, standardized radiographs were taken to monitor pen-implant bone levels. Results: All implants osseointegrated. A total bone loss of 0.48 +/- 0.66 mm was measured in the equicrestal INT group, 0.69 +/- 0.43 mm in the equicrestal EXT group, 0.79 +/- 0.93 mm in the subcrestal INT group, and 1.56 +/- 0.53 mm in the subcrestal EXT group (P>.05, paired t tests). Within the four groups, bone loss over time became significantly greater in the EXT groups than in the INT groups. The greatest bone loss was noted in the subcrestal EXT group. Conclusion: Within the limits of this animal study, it seems that even without prosthetic loading, different microgap configurations exhibit different patterns of bone loss during nonsubmerged healing. Subcrestal positioning of an external butt joint microgap may lead to faster radiographic bone loss. Int J Prosthodont 2011;24:445-452.

Influence of Microgap Location and Configuration on Peri-implant Bone Morphology in Nonsubmerged Implants: An Experimental Study in Dogs

Purpose: It is unknown whether different micro gap configurations can cause different pen-implant bone reactions. Therefore, this study sought to compare the peri-implant bone morphologies of two implant systems with different implant-abutment connections. Materials and Methods: Three months after mandibular tooth extractions in six mongrel dogs, two oxidized screw implants with an external-hex connection were inserted (hexed group) on one side, whereas on the contralateral side two grit-blasted screw implants with an internal Morse-taper connection (Morse group) were placed. on each side, one implant was inserted level with the bone (equicrestal) and the second implant was inserted 1.5 mm below the bony crest (subcrestal). Healing abutments were inserted immediately after implant placement. Three months later, the peri-implant bone levels, the first bone-to-implant contact points, and the width and steepness of the peri-implant bone defects were evaluated histometrically. Results: All 24 implants osseointegrated clinically and histologically. No statistically significant differences between the hexed group and Morse group were detected for either the vertical position for peri-implant bone levels (Morse equicrestal -0.16 mm, hexed equicrestal -0.22 mm, Morse subcrestal 1.50 mm, hexed subcrestal 0.94 mm) or for the first bone-to-implant contact points (Morse equicrestal -2.08 mm, hexed equicrestal -0.98 mm, Morse subcrestal -1.26 mm, hexed subcrestal -0.76 mm). For the parameters width (Morse equicrestal -0.15 mm, hexed equicrestal -0.59 mm, Morse subcrestal 0.28 mm, hexed subcrestal -0.70 mm) and steepness (Morse equicrestal 25.27 degree, hexed equicrestal 57.21 degree, Morse subcrestal 15.35 degree, hexed subcrestal 37.97 degree) of the pen-implant defect, highly significant differences were noted between the Morse group and the hexed group. Conclusion: Within the limits of this experiment, it can be concluded that different microgap configurations influence the size and shape of the peri-implant bone defect in nonsubmerged implants placed both at the crest and subcrestally. INT J ORAL MAXILLOFAC IMPLANTS 2010;25:540-547