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A comparative analysis of the microarchitecture of cortical membranous and cortical endochondral onlay bone grafts in the craniofacial skeleton.

Previous work in this laboratory established that an onlay bone graft's survival is determined primarily by its relative cortical and cancellous composition rather than its embryologic origin. A volumetric analysis of external bone graft resorption, however, does not explain the internal microarchitectural changes that may be occurring as these grafts become incorporated. To expand the knowledge of bone graft dynamics beyond volumetric parameters, a better understanding of the internal processes of bone graft remodeling is needed. In this comparative study of cortical onlay bone graft microarchitecture, the authors propose to show that cortical onlay bone grafts undergo measurable internal microarchitectural changes as they become incorporated into the surrounding craniofacial skeleton. In addition, the authors propose to further demonstrate similarities between the internal microarchitecture of cortical onlay bone grafts of different embryologic origin over time. Twenty-five adult New Zealand White rabbits were used for this study. They were divided into two groups of eight animals and one group of nine. The groups were killed at 3, 8, and 16 weeks. Cortical membranous and endochondral bone grafts were placed subperiosteally onto each rabbit's cranium. In addition, five ungrafted cortical endochondral and membranous bone specimens were used as controls. Microcomputed tomography (MCT) scanning and histomorphometric analysis were performed on all of the specimens to obtain detailed information regarding the microarchitecture of the cortical bone grafts. The parameters of bone volume fraction, bone surface area to volume, mean trabecular number, and anisotropy were used to give quantitative information about a bone's micro-organization. The results showed that there is no statistically significant difference between the cortical endochondral and the cortical membranous bone grafts for bone volume fraction, bone surface to volume, mean trabecular number, and anisotropy measurements for all time points. There were, however, statistically significant differences when comparing the control and 3-week groups to the 16-week group for all parameters. The advanced MCT technology and histomorphometric techniques proved to be effective in providing a qualitative and quantitative ultrastructural comparison of cortical endochondral and membranous onlay bone grafts over time. In this study, a statistically significant change in the internal microarchitecture of cortical onlay bone grafts of different embryologic origins was seen as they were remodeled and resorbed at all time points. Specifically, the onlay cortical bone grafts developed a less dense, more trabecular, and less organized internal ultrastructure. In addition, no difference in the three-dimensional ultrastructure of cortical endochondral and membranous bone was found. These results challenge some of the currently accepted theories of bone-graft dynamics and may eventually lead to a change in the way clinicians approach bone-graft selection for craniofacial surgery.

Animals↗

Anthropological measurements of the skeleton and parathyroid glands growth during fetal development.

The article presents the investigation of histomorphological differentiation and growth of parathyroid glands in human fetus from the second to the ninth lunar month. The longer and the shorter diameter of these glands were measured. The obtained values are compared with the development and the growth intensity of the skeleton (biparietal head diameter and femoral length of a fetus from lunar months 3 to 9) and with the role of the placenta in the mentioned processes. The results of our investigation show the concordance of the skeletal growth with the development and histomorphological differentiation of these glands. The factors involved in these processes point out the complex relationship between the mother and fetus during osteogenesis, expressed on the hormonal level, in mineral metabolism and placental activity.

Anthropometry↗

Tropomyosin isoform 5b is expressed in human erythrocytes: implications of tropomodulin-TM5 or tropomodulin-TM5b complexes in the protofilament and hexagonal organization of membrane skeletons.

The human erythrocyte membrane skeleton consists of hexagonal lattices with junctional complexes containing F-actin protofilaments of approximately 33-37 nm in length. We hypothesize that complexes formed by tropomodulin, a globular capping protein at the pointed end of actin filaments, and tropomyosin (TM), a rod-like molecule of approximately 33-35 nm, may contribute to the formation of protofilaments. We have previously cloned the human tropomodulin complementary DNA and identified human TM isoform 5 (hTM5), a product of the gamma-TM gene, as one of the major TM isoforms in erythrocytes. We now identify TM5b, a product of the alpha-TM gene, to be the second major TM isoform. TM5a, the alternatively spliced isoform of the alpha-TM gene, which differs by 1 exon and has a weaker actin-binding affinity, however, is not present. TM4, encoded by the delta-TM gene, is not present either. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, hTM5 comigrated with the slower TM major species in erythrocyte membranes, and hTM5b comigrated with the faster TM major species. TM5b, like TM5, binds strongly to tropomodulin, more so than other TM isoforms. The 2 major TM isoforms, therefore, share several common features: They have 248 residues, are approximately 33-35 nm long, and have high affinities toward F-actin and tropomodulin. These common features may be the key to the mechanism by which protofilaments are formed. Tropomodulin-TM5 or tropomodulin-TM5b complexes may stabilize F-actin in segments of approximately 33-37 nm during erythroid terminal differentiation and may, therefore, function as a molecular ruler. TM5 and TM5b further define the hexagonal geometry of the skeletal network and allow actin-regulatory functions of TMs to be modulated by tropomodulin. (Blood. 2000;95:1473-1480)

Amino Acid Sequence↗

Possible case of Rubinstein-Taybi syndrome in a prehistoric skeleton from west-central Illinois.

I describe an adult female skeleton (#Yo3-96) from the Yokem Mound skeletal series that had microcephaly; mild micrognathia; a broad nasal bridge; orbital hypertelorism; hypoplasia of the first metacarpals; pes planus; anomalies of the cervical spine, ribs, and sternum; and delayed epiphyseal remodeling of the radii and tibiae. Because the Yokem series has been subject to extensive measurement in previous studies, Yo3-96 can be characterized metrically and compared with others from her population. A differential diagnosis is made by comparing the available skeletal evidence with the known syndromes characterized by radial ray dysplasia. It is suggested that Yo3-96 represents the earliest known case of Rubinstein-Taybi syndrome, a mental retardation syndrome that has been described in individuals from several modern populations.

Adult↗

STR allelic frequencies in a German skeleton collection.

Chromosomal DNA was isolated from bones from a German skeleton collection (Goslar, 18th century) and detected by PCR. Nine microsatellite regions were amplified by multiplex reactions using the AmpFlSTR Profiler Plus kit and analysed to obtain their allelic distribution. A statistical evaluation of the results revealed no allelic differentiation between the historic sample and a modern German one at each locus.

Alleles↗

Trace analysis of endogenous and exogenous biomolecules from archaeological skeleton materials.

An analytical procedure was developed for the determination of pharmacologically active substances in archaeological skeleton materials. In comparative model studies, added ("spiked") test biomolecules of varying chemical behaviours were extracted from sample matrices and percentage of the analytes recovered were estimated using gas chromatography/mass spectrometry (GC/MS) or high performance liquid chromatography (HPLC). For the sterols and steroids studied, several organic solvents were appropriate. Extraction yields for the alkaloid nicotine, representing non-endogenous basic agents, were increased by alkalizing with triethylamine or by extracting with a two-phase system consisting of an alkaline aqueous and an organic layer (toluene). The flavonol quercetin was extractable only in an acidic environment. In a screening for native biomolecules using GC/MS, nicotine was identified in individual samples, in addition to lipophilic substances such as the endogenous cholesterol and its degradation products and two phytosterols which may have migrated into the bones from the surrounding soil.

Alkaloids↗

A case of Penicillium marneffei osteomyelitis involving the axial skeleton.

Fungal infection of bone by Penicillium marneffei is rare. We report on a case of Penicillium marneffei infection in a Filipino woman, which involved multiple soft-tissue abscesses and infection of the axial skeleton. Early diagnosis and treatment of this potentially reversible disease is emphasised. Such an approach is essential to prevent bony destruction from becoming too advanced and, more importantly, to prevent any damage to the spinal cord from occurring.

Adult↗

Prophylactic effect of BCG cell-wall skeleton on the tumor induction by 7,12-dimethylbenz[a]anthracene in mice: strain difference.

Prophylactic effect of repeated intravenous administrations of oil-attached BCG cell-wall skeleton (BCG-CWS) on the induction of tumor by 7,12-dimethylbenz[a]anthracene (DMBA) was investigated in various strains of mice. The subcutaneous injection of DMBA emulsified in oil induced squamous cell carcinoma in almost all of the strains of mice. Treatment of C57BL/6, BALB/c, and ddO strains with BCG-CWS with appropriate route and timing resulted in the retardation of DMBA-induced tumor development manifested by a prolonged latent period of tumor outgrowth. In contrast, the same BCG-CWS treatment of C3H/He and BTK mice was incapable in preventing such DMBA-induced carcinogenesis. Thus, the treatment with BCG-CWS was effective for preventing the DMBA-induced carcinogenesis in certain strains of mice, but the effectiveness varied depending on the strain. The implication of such a strain variationof the BCG-CWS effect on the prophylaxis of chemical carcinogenesis was discussed in the context of differences in the magnitude of immunopotentiation of the host by BCG-CWS.

9,10-Dimethyl-1,2-benzanthracene↗

Immunotherapy of human malignant melanoma with oil-attached BCG cell-wall skeleton.

A first case of malignant melanoma treated with oil-attached BCG cell-wall skeleton (BCG-CWS) is reported. The patient, a 70-year-old farmer, with metastatic malignant melanoma was treated intralesionally with oil-attached BCG-CWS or was given intradermally with oil-attached BCG-CWS together with irradiated autologous and allogeneic tumor cells. Four months after the start of the immunotherapy with oil-attached BCG-CWS, Primary tumor and metastatic inguinal lymph nodes regressed markedly without any significant complications. Lymphocytosis in the peripheral blood was usually observed after injection of oil-attached BCG-CWS.

Adjuvants, Immunologic↗

Effect of Nocardia and Mycobacterium cell-wall skeleton on autochthonous tumor grafts.

Cell-wall skeleton of Nocardia rubra and of Mycobacterium bovis BCG in the oil-attached form, when injected mixes with autografts of spontaneous mammary adenocarcinoma or of methylcholanthrene-induced sarcoma of mice, suppresses the growth of tumor autografts to a demonstrate extent. The BCG whole cell wall showed no effect. The local destruction of tumor autografts did not induce recognizable systemic immunity to the respective tumors. These findings are commended upon from immunological and chemical points of view.

Adenocarcinoma↗

Somatic and skeleton development of rat foetuses following in-utero exposure to isopropylantipyrine (propyphenazone) during the second trimester of gestation.

Isopropylantipyrine (IPA, propyphenazone) is a pyrazolone derivative, widely used as an antipyretic and analgesic drug. The aim of the study was to evaluate the influence of propyphenazone on rat development. IPA was administered to pregnant rats from day 8 to day 14 of pregnancy once a day, orally by a stomach tube at doses of 2.10 (R1), 21.0 (R2), and 210.0 mg/kg/day (R3). The dams were sacrificed on day 21 of gestation and corpora luteum, implants, resorptions, and live foetuses were counted. The weight of foetuses and placentas, the length of foetuses and their tails were checked. The foetuses were fixed in alcohol and skeletons were stained with alizarin. There was a statistical difference in body length in R1, R2 and numbers of subcutaneous ecchymose in R1. External and skeletal examination of the foetuses revealed no evidence of teratogenesis. It can be concluded that IPA has no harmful effects on the prenatal development of the rat offspring at doses used in the present study.

Animals↗

Morphological and biomechanical analysis of a skeleton from Roman imperial necropolis of Casalecchio di Reno (Bologna, Italy, II-III c. A. D.). A possible case of crutch use.

A Roman skeleton (T.130) from the roman necropolis of Casalecchio di Reno has been studied in order to understand if the hypothesis of crutch use, suggested by the severe articular degeneration at the hip joint that caused evident reduction of his locomotory possibilities, could be supported by the morphological alterations of other bones and joints. The pathological changes and muscular development of the upper limbs and shoulder girdle bones suggest that these parts were submitted to a great mechanical stress. The observations are consistent with the hypothesis of crutch use that would have involved a new weight-bearing function of the upper limbs in order to help locomotion, even though it is difficult to assess the number and type of the crutches. The comparison with other possible cases of crutch use reported in literature gives an additional support to the interpretation of the findings.

Biomechanical Phenomena↗

Automatic skeleton generation for visualizing 3D, time-dependent fluid flows: application to the virtual aneurysm.

Intracranial aneurysms are the primary cause of non-traumatic subarachnoid hemorrhage. Difficulties in identifying which aneurysms will grow and rupture arise because the physicians lack important anatomic and hemodynamic information. Through simulation, this data can be captured, but visualization of large simulated data sets becomes cumbersome, often resulting in visual clutter and ambiguity. To address these visualization issues, we developed an algorithm that extracts a skeleton of the patterns in 3D, time-dependent blood flow. The algorithm decomposes the blood flow into "bare-bones" components that can be visualized individually or superimposed together to formulate an understanding of the flow patterns in the aneurysm.

Cerebrovascular Circulation↗

Connective tissue skeleton in the normal left ventricle and in hypertensive left ventricular hypertrophy and chronic chagasic myocarditis.

Pictures certainly are worth a thousand words in the case of the structure of the connective tissue skeleton of normal and diseased myocardium. This report reviews the connective tissue matrix of the normal human myocardial tissue and the pathological myocardial fibrosis in left ventricular hypertrophy due to chronic arterial hypertension in humans and in human chronic chagasic myocarditis. The myocardial connective tissue matrix was studied employing a cell-maceration method that removes the myocardial tissue non-fibrous elements, and leaves behind a non-collapsed matrix, thus allowing a better three-dimensional view. Such information extends our knowledge of the expression of interstitial myocardial fibrous tissue in normal hearts and in hypertensive left ventricular hypertrophy and chronic chagasic myocarditis. The progressive accumulation of interstitial collagen fibers in both chronic cardiac diseases may be expected to decrease myocardial compliance and disrupt synchronous contractions of the ventricles during systole, contributing to a spectrum of ventricular dysfunction that involve either the diastolic or systolic phase of the cardiac cycle or both. In hypertensive heart disease myocardial fibrosis can be also implicated in the genesis of ventricular dysrhythmias, possible causes of sudden death among chronic hypertensive patients. Regarding chronic chagasic myocarditis, myocardial fibrosis is probably implicated in the genesis of malignant ventricular tachyarrhythmias (ventricular tachycardia and ventricular fibrillation), major causes of sudden death among patients with chronic Chagas' heart disease. The collagen distribution could interfere on the electrical properties of the myocardium. Fibrosis can block the cardiac impulse that may recycle (re-entry) through an alternative route and could slow conduction. In addition, the thick collagenous septa encompassing muscle fiber bundles could interfere with lateral impulse conduction, which would favor re-entry. Moreover, the methodology used is a useful tool to study the spatial organization of the collagen fibrils of the myocardium under normal and pathological conditions.

Chagas Cardiomyopathy↗

Mineral distribution in rat skeletons after exposure to a microgravity model.

Exposure to space flight models induces changes in the distribution of bone mineral in the human skeleton that has the features of a gravitational gradient. Regional bone mineral measurements with dual energy x-ray absorptiometry (DEXA) in male adults exposed to head-down tilt bed rest for 30 days show non-significant decrements in the pelvis and legs with 10% increases in the head region. Horizontal bed rest for 17 weeks reveals losses of bone mineral ranging from 2.2 to 10.4% from the lumbar spine to the calcaneus and an increase of 3.4% in the skull. Investigation of this phenomena would be most definitively carried out in an animal model. One candidate is the flight simulation model in the rat which removes body weight from the hind limbs and induces a cephalad fluid shift by suspending the animal by the tail. Weanling rats exposed to this model showed bone mineral to be lower in the hind limbs and higher in the skull after 3 weeks. These findings are similar in older 200 g animals after 2 weeks tail suspension. The purpose of this study was to determine the effect of age on the distribution of skeletal mineral in this model.

Aging↗

Spaceflight and the skeleton: lessons for the earthbound.

Loss of bone during extended space flight has long been a concern that could limit the ability of humans to explore the universe. Surprisingly the available data do not support the concept that weightlessness leads inexorably to a depleted skeleton unable to withstand the stress of a return to a 1g environment. Nevertheless, some bone loss does occur especially in those bones most stressed by gravity prior to flight, providing confirmation of the proposal formulated over a century ago by Julius Wolff that mechanical stress determines the form and function of bone. Although the phenomenon of bone loss with skeletal unloading, whether by space flight or immobilization or just taking a load off your feet (literally) is well established, the mechanisms by which bone senses load and adjusts to it are not so clear. What actually is the stimulus and what are the sensors? What are the target cells? How do the sensors communicate the message into the cells, and by what pathways do the cells respond? What is the role of endocrine factors versus paracrine or autocrine factors in mediating or modulating the response? None of these questions has been answered with certainty, but as will become apparent in this review, we have some clues directing us to the answers. Although the focus of this review concerns space flight, it seems highly likely that the mechanisms mediating the transmission of mechanical load to changes in bone formation and resorption apply equally well to all forms of disuse osteoporosis, and are likely to be the same mechanisms affected by other etiologies of osteoporosis.

Animals↗

Impact of antihypertensive therapy on the skeleton: effects of enalapril and AT1 receptor antagonist losartan in female rats.

No data are available about the effects of AT1 receptor antagonist losartan on the skeleton and there is also little information on the activity of an ACE inhibitor enalapril on bone metabolism. It is widely believed that the vasculature plays an important role in bone remodeling under normal and pathological conditions. We treated 14-week-old female Wistar rats with losartan, enalapril or saline. Administration of the ACE inhibitor enalapril and angiotensin II antagonist losartan had no effect on total malondialdehyde (MDA) in the blood and on urinary excretion of some eicosanoids and their metabolites. The administration of enalapril and losartan in a dose recommended for the treatment of hypertension did not cause significant changes in bone density, the ash and mineral content or morphometric parameters of the femur compared to the values found in control female rats.

6-Ketoprostaglandin F1 alpha↗

[The effects of electroporation-mediated erythropoietin (EPO) gene transfer into skeleton muscle on renal anemia].

OBJECTIVE: To investigate the effects of erythropoietin (EPO) gene transfer into skeleton muscle mediated by electroporation on renal anemia. METHODS: Renal failure models were created by adenine-excessive diet (150 mg per day). Plasmid vectors encoding EPO were transferred by electroporation after 80 days when mean blood urea nitrogen level (BUN) had increased from 3.4 mmol/L +/- 1.3 mmol/L to 18.1 mmol/L +/- 4.1 mmol/L and the hematocrit had decreased from 45.6% +/- 2.1% to 25.4% +/- 3.7%. During the process of treatment, adenine-excessive diet was given. Hb, HCT, BUN and Cre in blood were tested by automatic analyzer; EPO level in the serum was tested by EPO ELISA kit, EPO gene expression was proved by RT/PCR. The survival rate was calculated. RESULTS: Hematocrit increased to 34.4% +/- 7.5% only 7 days after the treatment and reached 91.4% of normal level (46% +/- 2%) after 5 weeks. The survival rate of test models after 9 weeks was 77.8%, which was remarkably higher than that of controls (16.7%). mRNA level of EPO gene expression was indicated by RT/PCR. CONCLUSION: Electroporation can increase the efficiency of EPO gene transfer and thus greatly improve hematocrit in mice and prolong the life-span of chronic renal anemia models. This method can provide a new way for treatment of EPO-responsive anemias.

Adenine↗