The impact of burial period on compact bone microstructure: Histological analysis of matrix loss and cell integrity in human bones exhumed from tropical soil.

Human bone histological analysis is a useful tool to assess post mortem diagenesis and to predict successful nuclear DNA typing of forensic material. This study is part of a series of studies developed by the authors intended to improve the understanding of post mortem diagenesis and to develop applications for DNA analysis of skeletal species from tropical soils, in order to optimize genetic and anthropological protocols. The aim of this study was to analyze the impact of burial period on the integrity of exhumed compact bone microstructure from tropical climate. In fragments of exhumed human femora from 39 individuals from the same cemetery (exhumed group) and 5 fresh femora from routine autopsies (control group), sections stained by hematoxylin-eosin were analyzed in order to measure bone microstructural integrity. We found that bone integrity index in exhumed group was negatively influenced by the period of burial (r = -0.37, p < 0.05) and highly significantly decreased (p < 0.0001) in comparison to control group. The period of burial and nitric acid decalcification time was positively correlated (r = 0.51; p < 0.01), leading to imply a bone petrification process during inhumation. Exhumed group showed higher level of matrix bone loss (p < 0.001), as expected, and 87% of cases analyzed were "tunneled" as described by Hackett. Bone integrity index and bone matrix tend to decrease in bones buried in tropical soil between 8-14 years of inhumation. This period is short if we consider cases in which there are preserved bones interred for longer periods in other environments. These data must be considered in cases where genetic identification of exhumed skeletons from tropical environment is required. The diagenesis in these bones and the variations of results found are discussed, clarifying some challenges for forensic laboratories, especially in DNA analysis.

Calcium orthophosphates (CaPO4): Occurrence and properties.

The present overview is intended to point the readers' attention to the important subject of calcium orthophosphates (CaPO4). This type of materials is of the special significance for the human beings because they represent the inorganic part of major normal (bones, teeth and antlers) and pathological (those appearing due to various diseases) calcified tissues of mammals. For example, atherosclerosis results in blood vessel blockage caused by a solid composite of cholesterol with CaPO4, while dental caries (tooth decay) and osteoporosis (a low bone mass with microarchitectural changes) mean a partial decalcification of teeth and bones, respectively, that results in replacement of a less soluble and harder biological apatite by more soluble and softer calcium hydrogenorthophosphates. Due to the compositional similarities to the calcified tissues of mammals, CaPO4 are widely used as biomaterials for bone grafting purposes. In addition, CaPO4 have many other applications. Thus, there is a great significance of CaPO4 for the humankind and, in this paper, an overview on the current knowledge on this subject is provided.

Risk of Malignancy in Thyroid Nodules with Non-Diagnostic Fine-Needle Aspiration: A Retrospective Cohort Study.

BACKGROUND: Fine-needle aspiration (FNA) of thyroid nodules is commonly performed, and despite the use of ultrasound (US) guidance, the rate of non-diagnostic FNAs is still significant. The risk of malignancy of thyroid nodules with a non-diagnostic FNA is not clearly defined. However, most studies exclude the majority of patients without a repeat biopsy or surgery, thus increasing the likelihood of selection bias. The aims of this study were to determine the malignancy risk in nodules with an initial non-diagnostic FNA, and to identify the factors associated with malignancy. METHODS: This was a retrospective cohort study of patients with thyroid nodules who underwent US-guided FNA between 2004 and 2010 and had a non-diagnostic result. Patients were followed until confirmatory diagnosis of the nature of the nodule was made. The outcome of malignant or benign disease was based on one of the following: (i) final surgical pathology following thyroidectomy; (ii) repeat biopsy; (iii) clinically, based on repeat ultrasound performed at least three years following biopsy; or (iv) report of thyroid status for patients without follow-up visits contacted by mail. RESULTS: There were 699 nodules from 665 patients included. The mean age was 59 ± 15 years, and 71.7% were women. There was complete follow-up of 495 nodules. After a median follow-up of 2.7 years, thyroid cancer was found in 15 nodules. The prevalence of malignancy was 3% (15/495). The presence of nodular calcifications was the strongest predictor of thyroid malignancy (odds ratio 5.03 [confidence interval 1.8-14.7]). Initial nodule size was inversely associated with malignancy (odds ratio 0.55 [confidence interval 0.28-0.93]). However, the 193 patients without follow-up had smaller nodules compared with those included in the analysis. None of the patients with repeatedly non-diagnostic results were diagnosed with thyroid cancer at follow-up. CONCLUSION: The prevalence of thyroid cancer in nodules with non-diagnostic results is lower than the malignancy rate in thyroid nodules in general, but not negligible. They should be followed as per guidelines with heightened suspicion for nodules containing calcifications. Nodules with repeatedly non-diagnostic FNA results especially in the absence of calcifications have a low risk of malignancy and may be observed.

Pediatric thyroid nodules: ultrasonographic characteristics and inter-observer variability in prediction of malignancy.

BACKGROUND: Pediatric thyroid nodules, while uncommon, have high malignancy risk. The objectives of the study were (1) to identify sonographic features predictive of malignancy; (2) to create a prediction model; and (3) to assess inter-observer agreement among radiologists. METHODS: All available cases of thyroid nodules, surgically removed between 2000 and 2009. Three radiologists reviewed the sonographic images; 2 pathologists reviewed the tissue specimens. Adult prediction models were applied. Interobserver variability was assessed. RESULTS: Twenty-seven subjects, mean age 13.1±3.4 years, were included. Nineteen nodules were differentiated thyroid carcinomas. On multivariate analysis, size was the only significant predictor of malignancy. On recursive partitioning analysis, size >35 mm with microcalcification and ill-defined margins yielded the best prediction model. Radiologist inter-observer agreement regarding malignancy was moderate (κ=0.50). CONCLUSIONS: Larger size, microcalcifications and ill-defined margins on ultrasound demonstrate the best predictive model for malignancy in the pediatric population. Experienced pediatric radiologists demonstrate moderate inter-observer agreement in prediction of malignancy.

Multimodal fundus imaging in choroidal osteoma.

PURPOSE: To describe the morphologic features of calcified and decalcified choroidal osteomas using multimodal imaging and correlate these findings with a previous histopathologic study. DESIGN: Retrospective observational case series. METHODS: Three patients with choroidal osteoma underwent complete ophthalmologic examination, fundus photography, and multimodal fundus imaging, including Fourier-domain optical coherence tomography (FD-OCT) and blue-light fundus autofluorescence (bAF). RESULTS: FD-OCT imaging of calcified tumors revealed a distinctive latticework pattern of reflectivity resembling the spongy bone structure seen histopathologically. On bAF the fluorescence was relatively well preserved overlying calcified tumors. In decalcified areas 2 patterns of reflectivity were identified: the first consisted of areas of relative hyperreflectivity with a lamellar appearance while the second was characterized by heterogeneous, hyperreflective, mound-like irregular areas associated with some posterior optical shadowing. Decalcified tumor areas had reduced overall fluorescence on bAF. CONCLUSION: FD-OCT demonstrated different reflectivity patterns in both calcified and decalcified portions of the choroidal osteoma, which may correspond to different stages of tumor evolution. A distinctive latticework pattern of reflectivity similar to spongy bone was seen in calcified tumors. These observations improve our knowledge of the in vivo structure of choroidal osteomas and may have implications for the diagnosis and management of this tumor.

Spectral-domain optical coherence tomography of the choroid in choroidal osteoma.

The authors report spectral-domain optical coherence tomography (SD-OCT) findings in a patient with decalcifying choroidal osteoma accompanied by a choroidal neovascular membrane and serous retinal detachment. A 13-year-old girl was found to have a choroidal osteoma in the left eye. The clinical diagnosis was confirmed by B-scan ultrasonography, computed tomography, and fluorescein and indocyanine green angiography. The SD-OCT findings over the decalcified portion included serous retinal detachment, photoreceptor outer segment disorganization, retinal pigment epithelial atrophy, deformed Bruch's membrane, and choroidal neovascular membrane. In contrast, the retinal structures over the calcified portion appeared to be preserved. SD-OCT showed loss of a vascular appearance and increased thickness in the affected choroid, especially in the decalcified portion. Choroidal thickening may be associated not only with choroidal osteomas, but also with tumor decalcification. These unique features on SD-OCT may be important in understanding poor visual prognosis when decalcification involves the fovea.

Optical coherence tomography of choroidal osteoma in 22 cases: evidence for photoreceptor atrophy over the decalcified portion of the tumor.

PURPOSE: To evaluate the retinal status overlying choroidal osteoma using optical coherence tomography (OCT). DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Twenty-two eyes with choroidal osteoma. METHODS: Choroidal osteoma was studied with fundus photography, ultrasonography, and OCT. MAIN OUTCOME MEASURE: Retinal status over the calcified and decalcified portions of choroidal osteoma. RESULTS: There were 8 completely calcified and 14 partially decalcified choroidal osteomas. Optical coherence tomography was performed over the calcified portion in 21 eyes and over the decalcified portion in 10 eyes. The calcified portion displayed an overlying intact inner retina (n = 21 [100%]), intact outer retina (n = 20 [95%]), and intact photoreceptor layer (n = 21 [100%]). In contrast, the decalcified portion showed an intact inner retina (n = 9 [90%]) and markedly thinned to absent outer retina and photoreceptor layers (n = 10 [100%]). Of the 18 eyes with subfoveolar choroidal osteoma, visual acuity (VA) was 20/20 to 20/50 in all 11 eyes with calcified tumor, and OCT confirmed preservation of retinal architecture. In contrast, VA was 20/200 or worse in all 7 eyes with subfoveolar decalcified tumor, correlating with OCT findings of outer retinal thinning and photoreceptor loss. CONCLUSIONS: The retina shows profound outer layer thinning and photoreceptor loss over decalcified choroidal osteoma. These findings correlate with poor VA over decalcified subfoveolar choroidal osteoma compared with good VA over calcified subfoveolar tumor.

Coeliac disease, unilateral occipital calcifications, and drug-resistant epilepsy: successful lesionectomy.

PURPOSE: To draw attention to the triad of coeliac disease (CD), occipital calcifications, and drug-resistant epilepsy, with focus on the outcome of epilepsy surgery. METHODS: We describe a male patient who despite a diagnosis of CD from the age of 9 did not comply with the gluten-free diet. At the age of 11 he developed simple and complex partial seizures with visual symptoms, anxiety, and ambulatory automatisms. His epilepsy appeared to be drug resistant, and after having tried nine antiepileptic drugs (AEDs), alone or in combinations, he underwent a presurgical evaluation at the age of 30. Interictal standard electroencephalograms (EEGs) disclosed frequent biparieto-occipital epileptiform discharges. Computed tomography showed cortical-subcortical punctate calcifications in the right parieto-occipital region, where his seizures seemed to start, according to ictal EEG registrations from intracranial strip electrodes. RESULTS: At the age of 31 he underwent epilepsy surgery. A 5 x 6 cm large area of the right parieto-occipital region was resected, including the area with calcifications. Except for a few short-lasting episodes of anxiety (simple partial seizures?) he has now been seizure-free for 12 years. AEDs were withdrawn 5 years ago. Postoperatively he was left with an upper left-sided quadrant anopsia, which is not bothering him. CONCLUSIONS: In patients with CD, unilateral occipital calcifications, and drug-resistant epilepsy, epilepsy surgery should be considered, as a lesionectomy might be very successful.

Observations on healing of human tooth extraction sockets implanted with bioabsorbable polylactic-polyglycolic acids (PLGA) copolymer root replicas: a clinical, radiographic, and histologic follow-up report of 8 cases.

OBJECTIVE: The objective was to conduct a clinical, radiographic, and histologic follow-up of alveolar socket healing in 8 human cases in which the extraction sockets of the involved teeth were treated with biodegradable root replicas before metallic implants were placed. STUDY DESIGN: Chair side prepared solid and porous forms of root replicas made out of polylactic-polyglycolic acids (PLGA) copolymer were utilized. Five patients were treated with the solid form and 3 with the porous form of the replicas. The cases were followed up at regular intervals postoperatively, and standardized photographs and radiographs were taken. The cylindrical core of biopsies that were removed with trephine for placement of titanium implants were processed and examined by light and transmission-electron microscopy. RESULTS: Both forms of the root replicas were well tolerated and biodegraded by the body. There were no histologically observable pathological tissue reactions at the time of implant application. However, the solid form seemed to cause an initial decalcification of the bone surrounding the extraction sockets that was subsequently repaired along with the bone healing of the extraction sockets. Such initial decalcification of the alveolar process was not observed in the cases that were treated with the porous form of root replicas. There was wide variation in the osseous component of the trephine-harvested biopsies in both treatment groups that suggests inconsistency in bone healing of the alveolar sockets. CONCLUSION: The 2 forms of root replicas under investigation were found to be biocompatible and biodegradable. But the compact solid form may cause an initial temporary lactic acid induced decalcification of the alveolar process, which makes it unsuitable for regular clinical application as compared to the granular porous form. The observed inconsistent and unpredictable bone regeneration calls for further research to develop more optimal replica materials.

Differential expression of IFN-gamma, IL-4, IL-10, and IL-1beta mRNAs in decalcified tissue sections of mouse lipopolysaccharide-induced periodontitis mandibles assessed by in situ hybridization.

To examine the role of T cell subgroups, Th1 and Th2, in the development of periodontitis, the expression of various cytokines was investigated in a mouse model of alveolar bone resorption using in situ hybridization (ISH) with digoxigenin-labeled oligonucleotides. When mice received repetitive injections with Escherichia coli lipopolysaccharide into the gingiva every 48 h, alveolar bone resorption was detectable after the fourth injection, reaching a maximum after the 13th injection. For the best performance of ISH, we first had to choose a decalcification protocol. Among various decalcification protocols, 10% EDTA (4 degrees C, 5-6 days) was the best for 28S rRNA staining. Positive cells for transcripts of interferon-gamma (Th1 product) were detected after the fourth injection, reaching a maximum after the tenth injection. A similar pattern was obtained for interleukin (IL)-10 mRNA (Th2 product) and IL-1beta, while the positive cell number reached a maximum after the 13th and 10th injections, respectively. The number of IL-4 mRNA (Th2 product)-positive cells remained low till the tenth injection, but increased thereafter. Consequently, we found that the population change from Th1 to Th2 in the inflammation site correlated with the transition from gingivitis to periodontitis, indicating differential roles of T cell subgroups in the development of periodontitis.

The rate of osteoclastic destruction of calcified tissues is inversely proportional to mineral density.

This study examined the relative ease with which three dissimilar mineralized tissues from one individual organ were resorbed by osteoclasts in vitro. Cells released from the long bones of prehatch chicks by agitating fragments of the chopped bones in medium were cultured for 24 hours on slices cut from an Elephas maximus molar so that enamel, dentine, and coronal cementum were present in bands on the surface of the slice. The resultant pits were measured using a video-rate, line-confocal reflection light microscope system. Variations in tissue mineralization were characterized by analysis of digital backscattered electron images. The enamel pits were smaller than the dentine and the cementum pits, but the dentine and cementum pits were not significantly different from each other. The sizes of the pits correlated with the relative mineral densities of the three tissues, showing that the rate of osteoclastic destruction of calcified tissues is inversely proportional to mineral density. This indicates that the initial step in osteoclasis, the removal of the mineral phase, determines the volume removed and is the rate-limiting step.

Fine structure of the human cochlear aqueduct: a light and transmission electron microscopic study of decalcified temporal bones.

The morphologic features of the human cochlear aqueduct were examined using both light and electron microscopy. The lumen of the cochlear aqueduct was observed to be filled with dense, irregular connective tissue corresponding to dura mater. At the entrance to the cerebrospinal fluid space, the dense connective tissue in the ductal lumen was covered with a thin layer of a few flattened cells, which was contiguous with the arachnoid membrane of the brain. A simple low cuboidal epithelium also separated the perilymphatic space from the lumen of the duct. Our observations confirm the presence of a barrier membrane at the opening to the perilymphatic space, and suggest that no transport occurs in the human cochlear aqueduct.

[Partial algodystrophy]. / Parzelläre Algodystrophie.

Two patients are described with parcellar algodystrophy, a special form of algodystrophy. Bone biopsy in one patient revealed increased bone resorption and irregular new bone formation. Parcellar algodystrophy is characterized by the very localized pathological changes in the region of a joint. Only a small area of demineralization may be visible on X-ray. Skeletal scintigraphy shows hyperfixation of technetium-99m already in an early stage. Although the inflammatory signs are clinically impressive, the ESR may be normal. History may reveal previous involved joints. Parcellar algodystrophy normally runs a benign course. The difficulty of correct diagnosis is discussed. The changes are compared to those of partial algodystrophy and algodystrophy with no radiological abnormality throughout the course of the disease.

Bone mineral analysis using single-energy computed tomography.

We present a new method for evaluating in vivo changes in bone mineralization in the peripheral skeleton, using computed tomography (CT). A set of bone mineralization indices are generated from numerous CT images of the patient's distal radius. The cross-sectional anatomy displayed by the CT scan allows for separate evaluation of the cortical and trabecular bone. Correction for possible drift of the CT number scale (Hounsfield scale) is achieved by scanning standard solutions of dipotassium hydrogen phosphate simultaneously with the forearm. Preliminary data indicate that this is a precise method for evaluating in vivo changes in bone mineralization.