Comparison of radiographic and tomographic evaluations for measurement of the Canal Flare Index in dogs / [Comparação entre avaliação radiográfica e tomográfica para mensuração do Canal Flare Index em cães]

The outcome of total hip arthroplasty (THA) in dogs is directly related to surgical planning. Templating of radiographs prior to THA should help the surgeon anticipate prosthesis size and femoral shape allowing canal fill of the proximal metaphysis by the implant ensuring primary stable fixation. The canal flare index (CFI) obtained from radiograph has been used as a measure of risk of complications for the technique in human beings and dogs. However, standard radiographs only provide limited data for the selection of cementless prostheses and the assessment of their fit within the femoral canal, due to factors like radiographic magnification and femoral rotation. Therefore, three-dimensional evaluation based on computed tomography (CT) may be a better tool for CFI measurement. The aim of this study was to compare anatomical measurement with CFI values obtained from craniocaudal radiography and CT. Craniocaudal radiographs using a horizontal radiographic beam (CR), CT, and anatomical macroscopic measurements (A) were obtained from 45 femurs from 23 canine cadavers. The differences between the values of CFI obtained from radiograph (CFI-R), computed tomography on transverse (CFI- TT) and longitudinal axis (CFI-TL) compared to the CFI obtained from macroscopic measurements - gold standard - (CFI-A), and 95% limits of agreement (LOA) between the values, were evaluated by the Bland-Altman method. Dimensions obtained from CT techniques had a greatest mean difference from anatomical and CFI values were also different (P=0.032). Under the experimental conditions, the craniocaudal radiograph, provided the most accurate measurement of the CFI (mean difference: 0.087 ± 0.42).(AU)

O resultado da artroplastia total do quadril (ATQ) em cães está diretamente relacionado ao planejamento cirúrgico. O templating radiográfico pré-operatório da ATQ deve ajudar o cirurgião a prever o tamanho da prótese e o formato do fêmur, o que permitirá um preenchimento ideal da metáfise proximal pelo implante, garantindo, assim, fixação primária estável. O índice de alargamento do canal (Canal Flare Index - CFI) obtido em radiografias tem sido utilizado como fator de risco de complicações para a técnica em humanos e cães. No entanto, as radiografias podem fornecer apenas dados limitados para a seleção de próteses não cimentadas e a avaliação do seu encaixe no canal femoral, devido a fatores como ampliação radiográfica e rotação femoral. Portanto, a avaliação tridimensional baseada na tomografia computadorizada (TC) pode ser uma ferramenta vantajosa para a mensuração do CFI. O objetivo deste estudo foi comparar a medida anatômica com os valores de CFI obtidos na radiografia craniocaudal e na TC. Radiografias craniocaudais utilizando feixe radiográfico horizontal (CR), tomografia computadorizada e medidas macroscópicas anatômicas (A) foram obtidas de 45 fêmures de 23 cadáveres caninos. As diferenças entre os valores de CFI obtidos na radiografia (CFI-R), na tomografia computadorizada no eixo transversal (CFI-TT) e no eixo longitudinal (CFI-TL), em comparação com os valores de CFI obtidos nas medições macroscópicas - padrão-ouro - (CFI-A) e os limites de concordância de 95% (LOA) entre os valores, foram avaliadas pelo método de Bland-Altman. As dimensões obtidas pelas técnicas de TC apresentaram maior diferença média dos valores anatômicos, e as do CFI também foram diferentes (P=0,032). Nas condições experimentais, a radiografia craniocaudal forneceu a medida mais precisa do CFI (diferença média: 0,087 ± 0,42) para representar o padrão-ouro deste estudo.(AU)

Double plating technique for fixing tibial plateau leveling osteotomy and modified cranial closing wedge ostectomy of the tibia in a dog with cranial cruciate ligament disease and excessive plateau angle: case report / [Técnica de placa dupla para fixação concomitante da osteotomia de nivelamento do platô tibial e ostectomia modificada em cunha de fechamento cranial da tíbia em um cão com doença do ligamento cruzado cranial e ângulo de platô tibial excessivo: relato de caso]

Tibial plateau leveling osteotomy (TPLO) associated to cranial wedge closing ostectomy (CCWO) has been one of the best options to manage cranial cruciate ligament (CCL) disease and excessive tibial plateau angle (TPA) in large dogs, however, the complication rate is potentially high. It is believed that a more robust fixation is necessary to stabilize them and decrease the risk of implant failure. A 6-year-old male American Pit Bull, weighing 36kg, with 90-day history of right hind limb lameness, was diagnosed with CCL disease. Due to the excessive tibial plateau angle (42°), TPLO was associated with a modified CCWO using a double plating technique. A final TPA of 12° was accomplished, and a restricted level of exercises and physiotherapy were recommended. The patient was followed monthly until the fifth month postoperatively, when radiographic bone consolidation and no lameness were observed. By the date of this submission, 3 years after the procedure, the owner has reported no complications. The double plating technique for fixing TPLO and modified CCWO proved to be effective for the treatment of CrCL deficiency in a large dog with an excessive TPA.(AU)

A osteotomia de nivelamento do platô tibial (TPLO) associada à ostectomia modificada em cunha de fechamento cranial da tíbia (CCWO) tem sido uma das melhores opções para tratamento de cães grandes com doença do ligamento cruzado cranial (DLCCr) e ângulo excessivo do platô tibial, mas o índice de complicações é alto. Acredita-se haver necessidade de fixação mais robusta para reduzir as chances de falha nos implantes. Um cão macho, seis anos, da raça American Pit Bull, 36kg, com histórico de claudicação em membro pélvico direito há 90 dias, foi diagnosticado com DLCCr. Devido ao ângulo excessivo do platô tibial (42°), a osteotomia de nivelamento do platô tibial foi associada à ostectomia modificada em cunha de fechamento cranial da tíbia (CCWO) por meio da técnica de placa dupla. No pós-operatório imediato, identificou-se TPA de 12°; exercícios controlados e fisioterapia foram recomendados e a evolução do quadro foi analisada mensalmente até o quinto mês pós-cirurgia. Com 150 dias de evolução, não houve alterações de locomoção e havia ocorrido completa consolidação radiográfica das osteotomias. Até o momento da submissão deste artigo, três anos pós-procedimento, o tutor relata ausência de complicações, via contato telefônico. Portanto, a técnica modificada mostrou-se eficaz no tratamento da DLCCr.(AU)

Biocompatibilidade do gel de quitosana associado ao glicerol fosfato na reparação de defeitos ósseos induzidos experimentalmente no rádio de coelhos (Oryctolagus cuniculus) / [Biocompatibility of chitosan gel associated with glycerol phosphate for the healing of experimentally induced radial defects in rabbits (Oryctolagus cuniculus)]

A engenharia de tecidos caracteriza-se como ciência interdisciplinar, a qual vem desenvolvendo biomateriais para a regeneração do tecido ósseo no âmbito das medicinas humana e veterinária. O objetivo desta pesquisa foi avaliar a regeneração óssea obtida da aplicação do hidrogel de quitosana associado ao glicerol fosfato em falha óssea experimentalmente induzida no rádio de coelhos. Foram utilizados 15 coelhos adultos, distribuídos aleatoriamente em dois grupos, representados por cada um dos rádios de cada animal, sendo um grupo tratado com hidrogel de quitosana associado ao glicerol fosfato (grupo biomaterial - GB) e um grupo que não recebeu tratamento com o biomaterial (grupo controle - GC). Os animais foram avaliados radiograficamente, por densitometria óptica e análise histológica, nos períodos 30, 60 e 90 dias pós-operatórios. Houve superioridade estatística na média geral das avaliações radiográficas do GB (2,33±0,48) sobre o GC (1,77±0,06). As médias gerais de avaliação densitométrica do GB foram superiores às do GC, sendo 6,207±1,374 e 5,71±1,512, respectivamente. A avaliação histopatológica do GB foi superior à do GC nos períodos de 30, 60 e 90 dias. Assim, é possível afirmar que o hidrogel de quitosana constitui biomaterial de características desejáveis, promovendo consolidação óssea mais rápida e eficiente, sem causar reações adversas.(AU)

Tissue engineering is an interdisciplinary science that has been developing biomaterials for bone regeneration in medicine and veterinary medicine, following an imminent need. The aim of this study was to evaluate bone regeneration after use of chitosan hydrogel associated with glycerol phosphate in experimentally induced bone gap in the radius of rabbits. Fifteen adult rabbits were randomly distributed in two experimental groups, represented by each radius of every single animal. The animals in the Biomaterial Group (GB) were treated with a glycerol phosphate-associated chitosan hydrogel and in the Control Group (GC) they received no treatment with the biomaterial. The animals were evaluated clinically, radiographically, histologically and by optic densitometry at 30, 60 and 90 days postoperatively. There was statistical superiority in the general average of the radiographic estimates of GB (2.33 ± 0.48) over the CG (1.77 ± 0.06). The general averages of GB densitometric evaluation were higher than the CG, being 6.207 ± 1.374 and 5.71 ± 1.512, respectively. Histopathological evaluation of GB was superior to CG in periods of 30, 60 and 90 days. Chitosan hydrogel constitutes a biomaterial of desired characteristics, promoting faster and more efficient bone repair when compared to GC.(AU)

Geographical patterns of allelic diversity in the Plasmodium falciparum malaria-vaccine candidate, merozoite surface protein-2.

The polymorphic merozoite surface protein-2 (MSP-2) of Plasmodium falciparum is a major malaria-vaccine candidate. In the present study, PCR and hybridization with allelic-specific probes were used to type the Msp-2 gene from isolates from hypo-endemic Brazil (N = 113), meso-endemic Vietnam (N = 208) and holo-endemic Tanzania (N = 67). The typing methods were designed to group isolates into the dimorphic allelic families FC27 and IC1 and to detect possible between-family recombination events. The analysis was complemented by a comparison of 156 Msp-2 sequences from the GenBank database with 12 additional sequences obtained during the present study. Statistically significant differences were detected in pair-wise comparisons of the distribution of Msp-2 allelic types in Brazil and Vietnam, and in Brazil and Tanzania, but not in Vietnam and Tanzania. The extent of allelic diversity in the Msp-2 gene, as estimated by the total number of different alleles found in a given parasite population and the mean multiplicity of infections, clearly paralleled the levels of malaria endemicity in the study areas. However, no correlation between age and multiplicity of infections was found in the subjects. The patterns of Msp-2 diversity in Brazil appeared to be temporally stable, since no significant difference was observed in the distribution of Msp-2 allelic types among isolates collected, 10--13 years apart, in the same area of Rondônia. Despite the extensive sequence diversity found in Msp-2 alleles, especially in the central repetitive region of the molecule, several instances of identical or nearly identical alleles were found among isolates from different countries and regions, possibly as a result of extensive homoplasy. No recombinant allele was detected by molecular typing in any of the study sites, and the GenBank database included only 12 recombinant sequences (representing 7% of all reported Msp-2 sequences), all of them with an IC1-type 5' end and an FC27-type 3' end. A single, putative, crossover site was characterised for all recombinant alleles. Most of the allelic diversity observed was therefore attributable to variation in the repetitive region of the gene, instead of recombination between alleles of dimorphic families (as commonly found, for example, in the Msp-1 gene). The implications of these findings for studies on the genetic and antigenic diversity of malarial parasites are discussed.

How prevalent is Plasmodium malariae in Rondônia, western Brazilian Amazon?

We have compared results of Plasmodium species identification obtained with conventional on-site microscopy of Giemsa-stained thick smears (GTS) and a semi-nested polymerase chain reaction (PCR) in 96 malaria patients from Rondônia, Western Brazilian Amazon. Mixed-species infections were detected by PCR in 30% patients, but no such case had been found on GTS. Moreover, P. malariae infections were detected in 9 of 96 patients (10%) by PCR, but were not identified by local microscopists. The potential impact of species misidentification on malaria treatment and control is discussed.

Allelic diversity and antibody recognition of Plasmodium falciparum merozoite surface protein 1 during hypoendemic malaria transmission in the Brazilian amazon region.

The polymorphic merozoite surface protein (MSP-1) of Plasmodium falciparum is a major asexual blood-stage malaria vaccine candidate. The impact of allelic diversity on recognition of MSP-1 during the immune response remains to be investigated in areas of hypoendemicity such as the Brazilian Amazon region. In this study, PCR was used to type variable regions, blocks 2, 4, and 10, of the msp-1 gene and to characterize major gene types (unique combinations of allelic types in variable blocks) in P. falciparum isolates collected across the Amazon basin over a period of 12 years. Twelve of the 24 possible gene types were found among 181 isolates, and 68 (38%) of them had more than one gene type. Temporal, but not spatial, variation was found in the distribution of MSP-1 gene types in the Amazon. Interestingly, some gene types occurred more frequently than expected from random assortment of allelic types in different blocks, as previously found in other areas of endemicity. We also compared the antibody recognition of polymorphic (block 2), dimorphic (block 6), and conserved (block 3) regions of MSP-1 in Amazonian malaria patients and clinically immune Africans, using a panel of recombinant peptides. Results were summarized as follows. (i) All blocks were targeted by naturally acquired cytophilic antibodies of the subclasses IgG1 and IgG3, but the balance between IgG1 and IgG3 depended on the subjects' cumulative exposure to malaria. (ii) The balance between IgG1 and IgG3 subclasses and the duration of antibody responses differed in relation to distinct MSP-1 peptides. (iii) Antibody responses to variable blocks 2 and 6 were predominantly type specific, but variant-specific antibodies that target isolate-specific repetitive motifs within block 2 were more frequent in Amazonian patients than in previously studied African populations.

Allelic diversity at the merozoite surface protein-1 (MSP-1) locus in natural Plasmodium falciparum populations: a brief overview.

The merozoite surface protein-1 (MSP-1) locus of Plasmodium falciparum codes for a major asexual blood-stage antigen currently proposed as a major malaria vaccine candidate. The protein, however, shows extensive polymorphism, which may compromise its use in sub-unit vaccines. Here we compare the patterns of allelic diversity at the MSP-1 locus in wild isolates from three epidemiologically distinct malaria-endemic areas: the hypoendemic southwestern Brazilian Amazon (n = 54), the mesoendemic southern Vietnam (n = 238) and the holoendemic northern Tanzania (n = 79). Fragments of the variable blocks 2, 4a, 4b and 6 or 10 of this single-copy gene were amplified by the polymerase chain reaction, and 24 MSP-1 gene types were defined as unique combinations of allelic types in each variable block. Ten different MSP-1 types were identified in Brazil, 23 in Vietnam and 13 in Tanzania. The proportion of genetically mixed infections (isolates with parasites carrying more than one MSP-1 version) ranged from 39% in Brazil to 44% in Vietnam and 60% in Tanzania. The vast majority (90%) of the typed parasite populations from Brazil and Tanzania belonged to the same seven most frequent MSP-1 gene types. In contrast, these seven gene types corresponded to only 61% of the typed parasite populations from Vietnam. Non-random associations were found between allelic types in blocks 4a and 6 among Vietnamese isolates, the same pattern being observed in independent studies performed in 1994, 1995 and 1996. These results suggest that MSP-1 is under selective pressure in the local parasite population. Nevertheless, the finding that similar MSP-1 type frequencies were found in 1994 and 1996 argues against the prominence of short-term frequency-dependent immune selection of MSP-1 polymorphisms. Non-random associations between MSP-1 allelic types, however, were not detected among isolates from Brazil and Tanzania. A preliminary analysis of the distribution of MSP-1 gene types per host among isolates from Tanzania, but not among those from Brazil and Vietnam, shows significant deviation from that expected under the null hypothesis of independent distribution of parasites carrying different gene types in the human hosts. Some epidemiological consequences of these findings are discussed.

Allelic diversity at the merozoite surface protein-1 locus of Plasmodium falciparum in clinical isolates from the southwestern Brazilian Amazon.

Nucleotide sequences of each variable block in the Plasmodium falciparum merozoite surface protein-1 gene (PfMSP-1) may be grouped into one of two or three possible allelic types, named after the reference isolates MAD20, K1, and RO33. Allelic diversity at this locus basically results from different combinations of allelic types in variable blocks. We used a polymerase chain reaction (PCR)-based strategy to type the variable blocks 2, 4a, 4b, and 10 of the PfMSP-1 gene of P. falciparum isolates from 54 symptomatic malaria patients living in Rondonia, a hypoendemic area in the southwestern Brazilian Amazon. Ten different PfMSP-1 gene types, defined as unique combinations of allelic types in variable blocks, were identified among the 54 isolates. Twenty-one isolates (39%) harbored more than one gene type and two had at least three genetically distinct clones. Hybrid sequences, with a MAD20-type sequence in the 5' segment (4a) and a K1-type sequence in the 3' segment (4b), were quite common in block 4. Direct sequencing of block 4 PCR products revealed a new putative recombination site in four isolates. In contrast with previous studies, the observed distribution of gene types does not deviate significantly from that expected under the null hypothesis of random association between allelic types detected in each variable block. These contradictory data are discussed with reference to the immunoepidemiologic features prevailing in distinct malaria-endemic areas.