RESUMO
Although three species of the genus Macrourus are recognized in the Southern Ocean, DNA sequencing of the mitochondrial COI gene revealed four well-supported clades. These barcode data suggest the presence of an undescribed species, a conclusion supported by meristic and morphometric examination of specimens.
Assuntos
Código de Barras de DNA Taxonômico , Gadiformes/genética , Animais , Complexo IV da Cadeia de Transporte de Elétrons/genética , Gadiformes/classificação , Especificidade da EspécieRESUMO
One of the rewards of having a Drosophila melanogaster whole-genome sequence will be the potential to understand the molecular bases for structural features of chromosomes that have been a long-standing puzzle. Analysis of 2.6 megabases of sequence from the tip of the X chromosome of Drosophila identifies 273 genes. Cloned DNAs from the characteristic bulbous structure at the tip of the X chromosome in the region of the broad complex display an unusual pattern of in situ hybridization. Sequence analysis revealed that this region comprises 154 kilobases of DNA flanked by 1.2-kilobases of inverted repeats, each composed of a 350-base pair satellite related element. Thus, some aspects of chromosome structure appear to be revealed directly within the DNA sequence itself.
Assuntos
Drosophila melanogaster/genética , Cromossomo X/genética , Animais , Bandeamento Cromossômico , Biologia Computacional , Cosmídeos , Elementos de DNA Transponíveis , DNA Satélite , Genes de Insetos , Hibridização In Situ , Sequências Repetitivas de Ácido Nucleico , Análise de Sequência de DNA , Cromossomo X/ultraestruturaRESUMO
Thioredoxin reductase (TrxR), a NADPH-dependent disulfide oxidoreductase, is vital in numerous cellular processes including defence against reactive oxygen species, cell proliferation and signal transduction. TrxRs occur in 2 forms, a high Mr enzyme characterized by those of mammals, the malaria parasite Plasmodium falciparum and some worms, and a low Mr form is present in bacteria, fungi, plants and some protozoan parasites. Our hypothesis is that the differences between the forms can be exploited in the development of selective inhibitors. In this study, cyclodextrin- and sulfonic acid-derived organotelluriums known to inhibit mammalian TrxR were investigated for their relative efficacy against P. falciparum TrxR (PfTrxR), a high Mr enzyme, and Trichomonas vaginalis TrxR (TvTrxR), a low Mr form of TrxR. The results suggest that selective inhibition of low Mr TrxRs is a feasible goal.
Assuntos
Antiprotozoários/farmacologia , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/enzimologia , Tiorredoxina Dissulfeto Redutase/metabolismo , Trichomonas vaginalis/efeitos dos fármacos , Trichomonas vaginalis/enzimologia , Animais , Ciclodextrinas/química , Humanos , Peso Molecular , Tiorredoxina Dissulfeto Redutase/químicaRESUMO
Exposure to blast overpressure (BOP) is associated with behavioral, cognitive, and neuroimaging abnormalities. We investigated the dynamic responses of cortical vasculature and its relation to microglia/macrophage activation in mice using intravital two-photon microscopy following mild blast exposure. We found that blast caused vascular dysfunction evidenced by microdomains of aberrant vascular permeability. Microglial/macrophage activation was specifically associated with these restricted microdomains, as evidenced by rapid microglial process retraction, increased ameboid morphology, and escape of blood-borne Q-dot tracers that were internalized in microglial/macrophage cell bodies and phagosome-like compartments. Microdomains of cortical vascular disruption and microglial/macrophage activation were also associated with aberrant tight junction morphology that was more prominent after repetitive (3×) blast exposure. Repetitive, but not single, BOPs also caused TNFα elevation two weeks post-blast. In addition, following a single BOP we found that aberrantly phosphorylated tau rapidly accumulated in perivascular domains, but cleared within four hours, suggesting it was removed from the perivascular area, degraded, and/or dephosphorylated. Taken together these findings argue that mild blast exposure causes an evolving CNS insult that is initiated by discrete disturbances of vascular function, thereby setting the stage for more protracted and more widespread neuroinflammatory responses.
Assuntos
Traumatismos por Explosões/patologia , Lesões Encefálicas/patologia , Macrófagos/patologia , Microglia/patologia , Animais , Barreira Hematoencefálica/patologia , Western Blotting , Encéfalo/irrigação sanguínea , Encéfalo/patologia , Modelos Animais de Doenças , Imunofluorescência , Imuno-Histoquímica , Microscopia Intravital , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microvasos/patologiaRESUMO
Brain cells manufacture and secrete angiogenic peptides after focal cerebral ischemia, but the purpose of this angiogenic response is unknown. Because the maximum possible regional cerebral blood flow is determined by the quantity of microvessels in each unit volume, it is possible that angiogenic peptides are secreted to generate new collateral channels; other possibilities include neuroprotection, recovery/regeneration, and removal of necrotic debris. If the brain attempts to create new collaterals, microvessel density should increase significantly after ischemia. Conversely, if angiogenic-signaling molecules serve some other purpose, microvessel densities may increase slightly or not at all. To clarify, the authors measured microvessel densities with quantitative morphometry. Left middle cerebral arteries of adult male Sprague-Dawley rats were occluded with intraluminal nylon suture for 4 hours followed by 7, 14, 19, or 30 days of reperfusion. Controls received no surgery or suture occlusion. Changes in microvessel density and macrophage numbers were measured by light microscopic morphometry using semiautomated stereologic methods. Microvessel density increased only in the ischemic margin adjacent to areas of pannecrosis and was always associated with increased numbers of macrophages. Ischemic brain areas without macrophages displayed no vascularity changes compared with normal animals. These data suggest that ischemia-induced microvessels are formed to facilitate macrophage infiltration and removal of necrotic brain.
Assuntos
Macrófagos/patologia , Neovascularização Patológica/patologia , Acidente Vascular Cerebral/patologia , Animais , Isquemia Encefálica/patologia , Humanos , Microcirculação/patologia , Modelos NeurológicosRESUMO
PURPOSE: This preclinical rat pilot study quantifies retinal microvessel, endothelial, and pericyte population changes produced by proton irradiation METHODS AND MATERIALS: The left eyes of rats were irradiated with single doses of 8, 14, 20, and 28 Gy protons; right eyes, with two fractions. Animals were euthanized, and eyes were removed; elastase digests were prepared, and cell populations were counted in sample fields. Results were compared with unirradiated controls. RESULTS: Progressive time- and dose-dependent endothelial cell loss occurred following all schedules. Cell loss was significantly different from control values (p < 0.001) following 28 Gy and following 20 Gy (p < 0.05) in a single dose. Endothelial cell loss was the same for single- and split-dose schedules. Progressive endothelial cell loss produced vessel collapse and acellular vessel strands. Endothelial cells were in the G(0) phase of the mitotic cycle. 28 Gy produced photoreceptor cell loss. CONCLUSION: The retinal digest is an elegant bioassay to quantify the microvessel population response. Single- and split-dose schedules appear to yield similar outcomes, in terms of endothelial cell density.
Assuntos
Endotélio Vascular/efeitos da radiação , Prótons , Vasos Retinianos/efeitos da radiação , Animais , Relação Dose-Resposta à Radiação , Endotélio Vascular/citologia , Microcirculação/efeitos da radiação , Projetos Piloto , Doses de Radiação , Ratos , Fase de Repouso do Ciclo Celular/efeitos da radiaçãoRESUMO
Mutations in the STM2 gene cause familial Alzheimer's disease (AD) in Volga Germans. To understand the function of this protein and how mutations lead to AD, it is important to determine which cell types in the brain express this gene. In situ hybridization histochemistry indicates that STM2 expression in the human brain is widespread and is primarily neuronal. In addition, STM2 mRNA is expressed in a cell line with neuronal origins. Quantification of the level of expression of the STM2 message in the basal forebrain, frontal cortex, and hippocampus reveals a significant decrease in AD-affected subjects compared to normal age-matched controls. These data suggest that downregulation of neuronal STM2 gene expression may be involved in the progression of AD.
Assuntos
Doença de Alzheimer/metabolismo , Encéfalo/metabolismo , Proteínas de Membrana/biossíntese , RNA Mensageiro/biossíntese , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Encéfalo/patologia , Linhagem Celular , Feminino , Regulação da Expressão Gênica , Humanos , Imuno-Histoquímica , Hibridização In Situ , Masculino , Proteínas de Membrana/genética , Pessoa de Meia-Idade , Presenilina-2RESUMO
Unfixed cryostat sections of rat liver were incubated to demonstrate D-amino acid oxidase activity at the ultrastructural level. Incubation was performed by mounting the sections on a semipermeable membrane which was stretched over a gelled incubation medium containing D-proline as substrate and cerium ions as capture reagent for hydrogen peroxide. After an incubation period of 30 min, ultrastructural morphology was retained to such an extent that the final reaction product could be localized in peroxisomes, whereas the crystalline core remained unstained. Control incubations were performed in the absence of substrate; the lack of final reaction product in peroxisomes indicated the specificity of the reaction. We conclude that the semipermeable membrane technique opens new perspectives for localization of enzyme activities at the ultrastructural level without prior tissue fixation, thus enabling localization of the activity of soluble and/or labile enzymes.
Assuntos
Crioultramicrotomia/métodos , D-Aminoácido Oxidase/metabolismo , Fígado/enzimologia , Microscopia Eletrônica/métodos , Animais , Fígado/ultraestrutura , Masculino , Ratos , Ratos WistarRESUMO
Cytochemical data in the literature reporting localization of sodium, potassium adenosine triphosphatase (Na(+), K(+)-ATPase) in the blood-brain barrier (BBB) have been contradictory. Whereas some studies showed the enzyme to be located exclusively on the abluminal endothelial plasma membrane, others demonstrated it on both the luminal and abluminal membranes. The influence of fixation on localization of the enzyme was not considered a critical factor, but our preliminary studies showed data to the contrary. We therefore quantitatively investigated the effect of commonly used fixatives on the localization pattern of the enzyme in adult rat cerebral microvessels. Fixation with 1%, 2%, and 4% formaldehyde allowed deposition of reaction product on both the luminal and abluminal plasma membranes. The luminal reaction was reduced with increasing concentration of formaldehyde. Glutaraldehyde at 0.1%, 0.25%, 0.5%, in combination with 2% formaldehyde, drastically inhibited the luminal reaction. The abluminal reaction was not significantly altered in all groups. These results show that luminal localization of BBB Na(+), K(+)-ATPase is strongly dependent on fixation. The lack of luminal localization, as reported in the literature, may have been the result of fixation. The currently accepted abluminal polarity of the enzyme should be viewed with caution.
Assuntos
Barreira Hematoencefálica , Encéfalo/enzimologia , Endotélio Vascular/enzimologia , ATPase Trocadora de Sódio-Potássio/análise , Fixação de Tecidos , Animais , Encéfalo/irrigação sanguínea , Capilares/enzimologia , Capilares/patologia , Endotélio Vascular/patologia , Fixadores , Formaldeído , Glutaral , Masculino , Ratos , Ratos Sprague-Dawley , Fixação de Tecidos/métodosRESUMO
In 50 patients receiving radical radiotherapy for carcinoma of the bladder a well-defined tumour was observed in the bladder either by trans-abdominal ultrasonography or by CT scanning prior to treatment. The histological type was transitional cell carcinoma 46, adenocarcinoma 1 and squamous cell carcinoma 3. The cross-sectional area of the tumour in the transverse plane was measured before the start of radiotherapy. In 37 patients serial measurements were made by ultrasonography during and after radical radiotherapy. In three patients no regression was seen during the first 14-27 days of treatment. In 34 patients with transitional cell carcinoma the median area-halving time was 37 days. Serial measurements of the volume of the tumour before, during and after radiotherapy were made by CT scanning in 29 patients. In two patients the tumour progressed with a volume-doubling time of 93 and 108 days, respectively. In 25 patients with transitional cell carcinoma the median volume-halving time was 29 days with a median area-halving time of 44 days. Linear-regression analysis of the initial tumour areas (31 patients) as measured by ultrasonography and CT scanning was performed. The correlation coefficient was r = 0.69. The correlation coefficient for the area halving-times measured both by ultrasonography and CT scanning was r = 0.53 (nine patients). Biphasic regression with a second slower phase with area-halving and volume-halving times of several hundred days was observed in six patients. Calcification of the tumour during and after radiotherapy was observed in several patients. No significant change in the CT number of the tumour was observed after radiotherapy. The median initial tumour volume as measured by CT scanning was 38 (7-211) cm3. The median initial tumour area as measured by ultrasonography was 16 (6-40) cm2. A tumour of median size with a median halving-time would take 120 days (4 months) to regress completely. Cystoscopy within 6 months of the start of radiotherapy is unlikely to be of value because many bladder tumours are continuing to regress during this period of time. Progression of the tumour during or after radiotherapy can be detected either by CT scanning or ultrasonography.
Assuntos
Carcinoma de Células de Transição/radioterapia , Radioterapia de Alta Energia/métodos , Neoplasias da Bexiga Urinária/radioterapia , Idoso , Carcinoma de Células de Transição/diagnóstico por imagem , Feminino , Humanos , Masculino , Dosagem Radioterapêutica , Tomografia Computadorizada por Raios X , Ultrassonografia , Neoplasias da Bexiga Urinária/diagnóstico por imagemRESUMO
Mutations in the presenilin genes PS1 and PS2 cause familial Alzheimer's disease (AD). In a previous study, we reported that PS2 mRNA levels are decreased in the hippocampus, frontal cortex and basal forebrain of subjects with late-onset sporadic AD. In this study, we examined whether this downregulation occurs as the disease progresses from mild to severe stages or whether downregulation of PS2 expression is an early event in AD. We used in situ hybridization histochemistry to quantify the level of expression of PS2 message in the hippocampus of normal subjects and subjects with mild, moderate or severe AD. Several regions of the hippocampus which are sequentially susceptible to AD neuropathology as the disease progresses in severity were analyzed. We demonstrate that specific downregulation of PS2 expression is as severe in subjects with mild AD as it is in subjects in late stages of the disease. In addition, we show that hippocampal regions that are relatively free of AD neuropathology during early stages of the disease exhibit severely compromised PS2 mRNA levels even in mild AD cases. In contrast, PS2 is expressed at normal levels in the cerebellum, a region which succumbs to significantly fewer AD-related insults even at very advanced stages of the disease. These results suggest that the specific downregulation of PS2 gene expression is an early event in sporadic late-onset AD.
Assuntos
Doença de Alzheimer/genética , Doença de Alzheimer/fisiopatologia , Proteínas de Membrana/genética , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Química Encefálica/genética , Cerebelo/fisiopatologia , Progressão da Doença , Feminino , Expressão Gênica/fisiologia , Hipocampo/fisiopatologia , Humanos , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , Presenilina-2 , RNA Mensageiro/análise , Índice de Gravidade de DoençaRESUMO
Two patients with cancer were evaluated in whom there was evidence for the simultaneous ectopic production of parathyroid hormone (PTH) and calcitonin (CT). One patient had a gastric carcinoid and the other had a pancreatic islet cell carcinoma. Abnormal concentrations of parathyroid hormone and calcitonin were demonstrated by radioimmunoassay in the peripheral blood of each patient and in the gastric tumor. In the pancreatic tumor, immunohistologic studies also demonstrated the presence of CT and PTH and suggested that each hormone was produced by different cells of the tumor. Plasma concentrations of the hormones responded to functional tests of secretion and to tumor chemotherapy. These studies demonstrate the simultaneous ectopic production of the two physiologically antagonistic hormones, PTH and CT, and confirm their value as diagnostic markers for several types of malignancies.
Assuntos
Adenoma de Células das Ilhotas Pancreáticas/metabolismo , Calcitonina/biossíntese , Tumor Carcinoide/metabolismo , Hormônios Ectópicos/biossíntese , Neoplasias Pancreáticas/metabolismo , Hormônio Paratireóideo/biossíntese , Neoplasias Gástricas/metabolismo , Adenoma de Células das Ilhotas Pancreáticas/análise , Adulto , Calcitonina/análise , Tumor Carcinoide/análise , Feminino , Hormônios Ectópicos/análise , Humanos , Neoplasias Hepáticas , Metástase Neoplásica , Neoplasias Pancreáticas/análise , Hormônio Paratireóideo/análise , Neoplasias Gástricas/análiseRESUMO
The effects of one week of estrogen replacement on choline acetyltransferase (ChAT) and trkA mRNA expression are examined in young and aged rodents to determine whether estrogen continues to affect cholinergic neurons in aging brain. Significant increases in ChAT and trkA are observed in the nucleus basalis of Meynert (nBM) of both age groups. ChAT expression is also increased in the HDB without changes in trkA expression. Results indicate modulation of ChAT expression by estrogen is retained in the aged rodent brain and suggests the possibility that changes in ChAT expression may be dissociated from concurrent alterations in trkA.
Assuntos
Envelhecimento/metabolismo , Proteínas de Transporte/genética , Colina O-Acetiltransferase/genética , Estradiol/farmacologia , Proteínas de Membrana/genética , Prosencéfalo/metabolismo , RNA Mensageiro/metabolismo , Receptor trkA , Animais , Feminino , Hibridização In Situ , Ovariectomia , Ratos , Ratos Endogâmicos F344RESUMO
Previous methods for determining morphological features of vascular networks in cerebral cortex were subject to arbitrary variation and bias. Unbiased estimates of vessel number, volume, surface area and length can be obtained using stereology but these techniques tend to be tedious and time-consuming. Stereologic protocols generally require micrographs that have to be analyzed manually for intersections of vessels on grid points or lines. In this report, we provide a simpler and more precise method for measuring morphological features of cerebral cortical microvessels. Images of microvessels in 1 microm toluidine blue stained sections were captured using a popular image analysis software package. Luminal surfaces of endothelial cells were automatically traced using commonly available features; the two-dimensional data of vessels (diameter, area, perimeter and number of vessels) were automatically computed and transferred to a spreadsheet. Three-dimensional features were then determined using basic stereologic equations. The method eliminates the need for manual measurements and is particularly time- and cost-effective for quantitative studies where numerous images have to be evaluated.
Assuntos
Córtex Cerebral/irrigação sanguínea , Processamento de Imagem Assistida por Computador , Neurociências/métodos , Animais , Vasos Sanguíneos/anatomia & histologia , Masculino , Microcirculação , Ratos , Ratos Sprague-DawleyRESUMO
The purpose of this study was to determine if proton irradiation can increase the localization of radiolabeled monoclonal antibodies (MAb) in subcutaneous (s.c.) or intracranial (i.c.) human lung tumors xenotransplanted in athymic rats. Rats with carcinoembryonic antigen (CEA)-expressing (NCI-H441) tumors were irradiated using 3 different proton time-dose regimens, followed by 111In-ZCE025, an anti-CEA MAb, which was injected 2 hr after the last dose of irradiation, and the animals were euthanized 3 days later for biodistribution and other assays. Proton irradiation at 10 gray (Gy) as a single dose or in 2 Gy fractions given on 5 consecutive days increased the uptake of 111In-ZCE025 into s.c. tumors by 292% and 182%, respectively, compared to nonirradiated controls. No enhancement in radiolabeled MAb delivery was seen after hemibrain irradiation in animals with i.c. tumors. Histopathological examination of both implantation sites showed a viable poorly differentiated adenocarcinoma with a decrease in blood vessel density, a decrease in mitotic activity, and an increase in areas of necrosis following irradiation as compared with adjacent nonirradiated tissue. CEA expression was generally maintained in vivo in that the marker was detectable in the tumor, plasma, and cerebrospinal fluid. Oxygen radical production by peripheral blood cells from s.c. and i.c. tumor-bearing rats exhibited strikingly different patterns of responsiveness. I.c. injected animals were 24% lighter than their s.c. injected counterparts, but no neurological signs of tumor progression were noted. The results indicate that proton irradiation can be used effectively to increase the delivery of radiolabeled MAb to s.c. implanted human lung tumor xenografts. However, in order to accomplish this in the brain, other radiation time-dose schedules and treatments may be needed.
Assuntos
Adenocarcinoma/radioterapia , Anticorpos Monoclonais , Neoplasias Encefálicas/radioterapia , Neoplasias Pulmonares/radioterapia , Neoplasias Pulmonares/secundário , Adenocarcinoma/diagnóstico por imagem , Adenocarcinoma/patologia , Adenocarcinoma/secundário , Animais , Peso Corporal , Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/patologia , Antígeno Carcinoembrionário/análise , Antígeno Carcinoembrionário/biossíntese , Antígeno Carcinoembrionário/imunologia , Linhagem Celular , Relação Dose-Resposta à Radiação , Humanos , Radioisótopos de Índio/farmacocinética , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/patologia , Masculino , Necrose , Tamanho do Órgão , Fagócitos/metabolismo , Projetos Piloto , Prótons , Radioimunoterapia , Cintilografia , Ratos , Ratos Nus , Explosão Respiratória , Baço/patologia , Superóxidos/análise , Superóxidos/metabolismo , Distribuição Tecidual , Transplante Heterólogo , Células Tumorais CultivadasRESUMO
Patients with germ cell testicular tumours customarily have repeated follow-up chest radiographs after treatment. This study assesses the contribution of chest radiography to the detection of recurrent disease in 162 patients. Six patients developed an intrathoracic recurrence, but in only one case was the chest radiography the only indication of recurrence. Five had other evidence to suggest recurrence, such as raised serum markers or palpable masses. The yield from follow-up chest radiographs in patients with germ cell testicular tumours is very low and their use must be balanced against both the harmful effects of radiation and the financial cost.
Assuntos
Disgerminoma/diagnóstico por imagem , Teratoma/diagnóstico por imagem , Neoplasias Testiculares/diagnóstico por imagem , Neoplasias Torácicas/diagnóstico por imagem , Adolescente , Adulto , Idoso , Disgerminoma/secundário , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Radiografia , Teratoma/secundário , Neoplasias Torácicas/secundárioRESUMO
A wound stabilizing effect of expanded polytetrafluoroethylene (ePTFE) membranes was evaluated in supra-alveolar periodontal defects in 5 beagle dogs. The defects, 5 to 6 mm in height, were surgically created around the 2nd, 3rd, and 4th mandibular premolar teeth in contralateral jaw quadrants. The root surfaces were conditioned with heparin, which, in this model, has been demonstrated to compromise periodontal healing and result in formation of a long junctional epithelium. Wound closure included application of ePTFE membranes around each premolar tooth in one jaw quadrant in each dog and flap positioning coronal to the cemento-enamel junction in both jaw quadrants. Healing progressed uneventfully except for 3 teeth in 2 dogs, which experienced membrane exposure. The dogs were sacrificed after a 4-week healing period and tissue blocks were prepared for histometric analysis. Connective tissue repair in heparin+membrane-treated teeth averaged 98% of the defect height compared to 84% in control heparin-treated teeth (P < or = 0.05). Junctional epithelium formation was smaller in membrane-treated teeth than in control teeth (P < or = 0.05) and was usually terminated coronal to the membrane. Bone regeneration was enhanced in membrane-treated teeth compared to controls (P < or = 0.01) and was strongly correlated to the area under the membrane in teeth without membrane exposure (r2 = 0.993; P = 0.002). This correlation was reduced when teeth with membrane exposure were included in the analysis (P < or = 0.05). Cementum regeneration was minimal under both treatment conditions. Root resorption was increased in membrane-treated compared to control teeth (P < or = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Regeneração Tecidual Guiada Periodontal , Membranas Artificiais , Doenças Periodontais/cirurgia , Politetrafluoretileno , Processo Alveolar/patologia , Processo Alveolar/fisiopatologia , Animais , Regeneração Óssea , Tecido Conjuntivo/patologia , Tecido Conjuntivo/fisiopatologia , Cemento Dentário/patologia , Cemento Dentário/fisiopatologia , Cães , Epitélio/patologia , Epitélio/fisiopatologia , Masculino , Doenças Periodontais/patologia , Doenças Periodontais/fisiopatologia , Periodonto/patologia , Periodonto/fisiopatologia , Regeneração , Reabsorção da Raiz/patologia , Reabsorção da Raiz/fisiopatologia , CicatrizaçãoRESUMO
It is often assumed that there is a direct relationship between the bone density adjacent to an implant, as revealed by radiographs, and the percent histologic osseointegration. Moreover, the lack of standardized methods for evaluation of histologic preparations makes it difficult to compare published studies, especially as little is known about the variables that influence these measurements. In this animal study, computer-assisted lineal analysis was used to evaluate the effects of subject, tooth position, and implant surface site on measured bone density and osseointegration in a bone augmentation experiment. Three sites--coronal lingual, apical lingual, and apical facial--were analyzed around each of 6 (3.75 x 8 mm) threaded machined titanium implants, as well as the apical facial site of 21 other implants placed in the mandibular premolar area of 5 dogs. In all sites, a progressive decrease in bone density was observed from bone adjacent to the implant to that at the titanium implant surface. There was an animal effect on osseointegration, but there were no differences between the mandibular premolar locations (second, third, and fourth). Most importantly, there were significant measurable effects attributable to the surface site examined. The need for carefully standardized histologic evaluations is established.
Assuntos
Perda do Osso Alveolar/etiologia , Implantes Dentários , Implantes Experimentais , Osseointegração , Perda do Osso Alveolar/diagnóstico por imagem , Análise de Variância , Animais , Densidade Óssea , Implantação Dentária Endóssea , Implantes Dentários/efeitos adversos , Cães , Implantes Experimentais/efeitos adversos , Mandíbula/diagnóstico por imagem , Mandíbula/patologia , Mandíbula/cirurgia , Radiografia , Estatísticas não ParamétricasRESUMO
Computer-assisted planimetry, computer-assisted lineal analysis, and point-counting stereology have been compared with respect to their reproducibility and the time required to analyze bone-implant integration. Sections of 6 threaded dental implants selected from a bone augmentation experiment for their wide range of new bone formation were analyzed by each method 3 times. The bone density and percentage of osseous integration were evaluated at 4 sites around each implant section. It was found that computer-assisted planimetry demonstrated a modest but significantly greater variance (P < .05) in bone density estimates when compared to the computer-assisted lineal analysis and point-counting methods. Computer-assisted planimetry requires a different method of measuring each parameter and separate fields of view to evaluate fields distant from the implant. However, this can all be accomplished with line probes, as in computer-assisted lineal analysis, which extend from the implant surface into the surrounding alveolar bone. Whereas computer-assisted planimetry requires a separate identification of the perimeter of each field to be analyzed (next to and distant from the implant), computer-assisted lineal analysis allows expansion of the field to be evaluated without creating a new field of view. Also, following a limited learning curve, both point-counting and computer-assisted lineal analysis required less time to complete than did computer-assisted planimetry.
Assuntos
Implantação Dentária Endóssea , Implantes Dentários , Mandíbula/patologia , Osseointegração , Processo Alveolar/patologia , Aumento do Rebordo Alveolar , Animais , Densidade Óssea , Cães , Estudos de Avaliação como Assunto , Processamento de Imagem Assistida por Computador/métodos , Mandíbula/cirurgia , Osteogênese , Reprodutibilidade dos Testes , Propriedades de SuperfícieRESUMO
We have reviewed the morphologic, electrophysiologic, biochemical, and functional methods of evaluating PN regeneration in animal models. There are a large number of anatomic techniques that can provide clear insights into the processes of peripheral nerve regeneration. Since many of these are costly in terms of labor, careful selection of the technique appropriate for the question asked is important. Two of the more important questions are: 1) What are the neurotrophic factors produced by the distal segment that attract the growing axon tip? and 2) What are the components of the basal lamina that facilitate the directed growth of the axons? To answer these questions, whole mount preparations provide the means to economically evaluate the result of experimental manipulation of the environment. Automated nerve fiber counts will be increasingly used to help interpret electrophysiologic studies. Quantitative as well as descriptive ultrastructural analyses will continue to provide valuable data that will be needed in the interpretation of biochemical and histochemical studies. Immunohistochemical probes are sure to become more important as the range of their specificities broadens. With the diversity of anatomic methods available and their capacity to help us visualize the processes occurring during nerve regeneration they will remain a key tool in these studies. Electrophysiologic methods that integrate the CAP and correlate it with the number of functioning NF are most useful. Functional methods are beginning to become more objective and quantitative. The most precise measurements are muscle weight and the isometric response of muscle to tetanic contraction. Sensory function has now been measured objectively by Horch. Single methods of measuring PN regeneration give only limited data, but by combining methods a better understanding of PN regeneration is possible. While understanding the limitations of each method and technique, multi-parameter animal models may provide data most helpful clinically. However, because of great species variability in the reparative response, caution must be given not to extrapolate too much from animal studies. We urge investigators to use the most objective methods available to measure nerve regeneration. Recognizing these limitations, however, animal studies will continue to provide significant insights into PN regeneration and should point the way to improved clinical practice.