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PubMedDate not supplied

On the influence of altered gravity on the growth of fish inner ear otoliths.

Inner ear stones (otoliths) of developing cichlid fish (Oreochromis mossambicus) were marked with the calcium tracer alizarin-complexone (AC) at 1g-earth gravity before and after a longterm (20 days) stay of the animals at moderate hypergravity conditions (3g; centrifuge). AC deposition at the otoliths resulted in two fluorescence bands, which enclosed the area grown during exposure to altered gravity. This area was measured with regard to size and asymmetry (size difference between the left and the right stones). Both utricular and saccular otoliths (lapilli and sagittae, respectively) were significantly smaller after hyper-g exposure as compared to parallely raised 1 g-control specimens. The asymmetry concerning the lapilli was pronouncedly decreased in comparison to the 1g-controls. These findings suggest, that the growth and the development of bilateral asymmetry of otoliths is guided by the environmental gravity vector. Some of the hyper-g animals revealed a kinetotic behaviour at the transfer from hyper-g to normal 1g-earth gravity conditions, which was qualitatively similar to the behaviour observed in previous experiments at the transfer from 1 g to microgravity in the course of parabolic aircraft flights. The lapillar asymmetry of kinetotic samples was found to be significantly higher than that of normally behaving experimental specimens. This result supports an earlier theoretical concept, according to which human static space sickness might be based on asymmetric utricular otoliths

Animals↗
PubMed2002

Neuronal feedback between brain and inner ear for growth of otoliths in fish.

Previous investigations revealed that fish inner ear otolith growth (concerning otolith size and calcium-incorporation) depends on the amplitude and the direction of gravity, suggesting the existence of a (negative) feedback mechanism. In search for the regulating unit, the vestibular nerve was unilaterally transected in neonate swordtail fish (Xiphophorus helleri) which were subsequently incubated in the calcium-tracer alizarin-complexone. Calcium incorporation ceased on the transected head sides, indicating that calcium uptake is neurally regulated.

Animals↗
PubMed2002

Influence of hypergravity on fish inner ear otoliths: II. Incorporation of calcium and kinetotic behaviour.

Larval siblings of cichlid fish (Oreochromis mossambicus) were subjected to hypergravity (hg; 3 g, 14 days) during development. Following the transfer to 1 g (i.e., stopping the centrifuge) they were separated into normally and kinetotically swimming individuals (the latter performed spinning movements). During hg, the animals were maintained in aquarium water containing alizarin-complexone (AC), a fluorescent calcium tracer. Densitometric measurements of AC uptake into inner ear otoliths (optical density of AC/micrometers2) revealed that the kinetotic individuals had incorporated significantly more AC/calcium than the normally behaving fish. Since the amount of otolithic calcium can be taken as an approximation for otolith weight, the present results indicate that the otoliths of kinetotically swimming samples were heavier than those of the normally behaving larvae, thus exhibiting a higher absolute weight asymmetry of the otoliths between the right vs. the left side of the body. This supports an earlier concept according to which otolith (or statolith) asymmetry is the cause for kinetoses such as human static space sickness.

Animals↗
PubMed2002

Influence of hypergravity on fish inner ear otoliths: I. Developmental growth profile.

Inner ear stones (otoliths) of larval cichlid fish Oreochromis mossambicus were marked with the calcium-tracer alizarin-complexone (AC) at 1 g earth gravity before and after a 3, 7, 14 or 21 days stay of the animals at hypergravity conditions (hg; 3 g, centrifuge). After the experiment, the otoliths' area between the two AC-labellings was measured with regard to size and asymmetry (size difference between the left and the right stones). Both utricular and saccular otoliths (lapilli and sagittae, respectively) continued growing in a linear way at hg, but growth was significantly slowed down as compared to parallely raised 1 g-control specimens. In case of bilateral asymmetry between the corresponding otoliths its formation in hg-animals became reduced as compared to the 1 g controls. The reduction of asymmetry was much more pronounced in the sagittae than in the lapilli. The latter result supports an earlier hypothesis, according to which especially a low sagittal asymmetry has a functional advantage. In general, the results strongly suggest that otolith growth is continuously regulated in dependence of the environmental gravity vector.

Animals↗
PubMed1999

Interference of magnetic resonance imaging contrast agents with the serum calcium measurement technique using colorimetric reagents.

A possible interaction between either linear (Gd-DTPA-BMA and Gd-DTPA) or macrocyclic (Gd-DOTA) gadolinium complexes used as magnetic resonance imaging (MRI) contrast agents and colorimetric technique reagents for the measurement of serum calcium was evaluated on human serum pools, and its mechanism was investigated by means of UV spectrometry and electro-spray ionization mass spectrometry (ESI-MS). The highest concentration tested was 2.5 mM (corresponding to a putative strictly intravascular distribution of the compound) and the lowest dose was 0.2 mM (i.e. about two elimination half lives). Serum calcium was dosed in duplicate by conventional colorimetric techniques involving o-cresol-phthalein complexone (OCP) or methylthymol blue (MTB) as reagents. No interference was detected when mixing Gd DOTA with serum, whatever the concentration. Gd DTPA (2.5 mM) did not interfere with the colorimetric technique either. Conversely, the Gd DTPA-BMA solution induced a concentration-related variation in apparent calcium levels. In the UV experiments, solutions of 2.5 mM MRI contrast media were mixed with OCP or MTB and UV absorption spectra were recorded between 400 and 800 nm. For Gd-DOTA/OCP and Gd-DOTA/MTB, no significant variations in the absorbance were detected. However, in the presence of Gd DTPA BMA, the absorbance of OCP and MTB showed substantial and immediate variations over time. The ESI-MS studies showed a complete displacement of Gd3+ ion in the case of Gd-DTPA BMA. In the presence of OCP, we observed the disappearance of Gd-DTPA BMA and the formation of the free ligand DTPA BMA and a new complex Gd OCP with an original stoichiometry of 2/2. Such a phenomenon did not occur in the case of Gd DOTA and Gd DTPA. The decomplexation of Gd-DTPA BMA in the presence of OCP can probably be explained by the weaker thermodynamic stability of Gd-DTPA BMA compared to that of Gd-DOTA and Gd DTPA.

Bromthymol Blue↗
PubMed1996

Effects of buccal shields on the maxillary dentoalveolar structures and the midpalatal suture--histologic and biometric studies in rabbits.

The mechanism of the buccal shields with regard to regulating the transversal relationship of the maxillary dental arch is uncertain. The periosteal pull theory, growth at the midpalatal suture and changes in equilibrium have all been proposed as explanatory factors. The aim of the study was to investigate the transversal development of the dental arch and the osseous remodeling at the lateral surfaces of the maxillary alveolar process and in the midpalatal suture after stretching the bucca (cheek) with buccal shields in the vestibulum. Ten New Zealand white male rabbits 12 weeks old were used. The animals were divided into two groups (control and experimental). Initial and final impression had been taken from the two groups and plaster models were made using biometric analysis in all animals of each group. Transversal measurements were taken of the distances between the maxillary right and left teeth. Tetracycline hydrochloride and Alizarin complexone were used to label the mineralizing tissues. After the animals had been killed, the biopsies from the midpalatal suture, the alveolar wall, and the periosteum were studied under a microscope. The results from the biometric analysis showed maxillary dental arch expansion in the two groups that was significantly larger in the experimental group. Microscopically, the growth at the midpalatal suture was also significantly larger in the experimental group, while no significant differences were found at the maxillary alveolar bone in the two groups. The conclusions drawn from this study are that the vestibular buccal shields bring about an increased expansion of the maxillary dental arch and increased growth at the midpalatal suture. No increased bone deposition was found at the buccal dentoalveolar wall, inducing doubts as to the accuracy of the periosteal pull theory for widening of the dental arch.

Activator Appliances↗
PubMed1998

Effects of static magnetic field on bone formation of rat femurs.

Effects of static magnetic fields (SMF) on bone formation of rat femurs, were evaluated using tapered rods made of magnetized and unmagnetized samarium cobalt of the same size. They were implanted transcortically into the middle diaphysis of rat femurs under press-fit loading. The bone mineral density (BMD) and bone calcium content were measured 12 weeks after implantation by dual-energy X-ray absorptiometry and chemical analysis with o-cresolphthalein complexon, respectively. The result revealed that the femurs adjacent to magnetized specimens had significantly higher BMD and calcium content than those adjacent to the unmagnetized specimen (p < 0.01). However, the value of BMD and calcium content of rats with magnetized specimens was similar to that of non-operated rats. No specific change was found in the body weight, serum Ca, activity of alkaline phosphatase, hemogram, and BMD of the tibia and humerus among the magnetized and unmagnetized. These results suggest that the long-term local SMF stimulation on the bone has a local effect to prevent the decrease in BMD caused by surgical invasion or implantation.

Absorptiometry, Photon↗
PubMed2006

Sensitivity and specificity improvement of an ion sensitive field effect transistors-based biosensor for potato glycoalkaloids detection.

Butyryl cholinesterase of different origin along with variations of the time of enzyme immobilization on the potentiometric transducer surface is offered to control the ion sensitive field effect transistor (ISFET)-based biosensor sensitivity. Because butyryl cholinesterase has been already used to develop the sensors for heavy metals, organophosphorus/carbamate pesticides, and steroidal glycoalkaloids analysis, the present study has been focused on the investigation and adjustment of the ISFET-based biosensor specificity exclusively to the glycoalkaloids. Utilization of ethylendiaminetetracetate (a complexon of heavy metal ions) and phosphotriesterase (a highly efficient catalyst for the hydrolysis of organophosphorus compounds) enabled the highly specific determination of glycoalkaloids at the background of lead and mercury (up to 500 microM of ions concentration) and paraoxon (up to 100 microM of pesticide concentration). The developed biosensor has been validated for glycoalkaloids detection in potato varieties cultivated in Ukraine, and the results obtained are compared to those measured by the methods of HPLC and TLC.

Alkaloids↗
PubMed2000

Healing of titanium implants in onlay bone grafts: an experimental rabbit model.

An experimental rabbit bone graft model for the study of bone formation and remodeling around titanium implants is described. A 2.5-cm long radius bone segment served as an onlay graft. Two commercially pure (c.p.) titanium implants were inserted into the bone graft prior to fixation to the inferior border of the mandibular base with osteosynthesis titanium screws. Each animal was operated twice, allowing follow-up periods of 6 weeks on one side and 6 months on the contralateral side. In order to study bone remodeling by means of fluoroscopy the animals received single injections of tetracyclin and alizarine complexone 2 weeks and 1 week, respectively, prior to sacrifice by perfusion fixation with glutaraldehyde. The bone and implants were excized en bloc, postfixed and embedded in plastic resin. Stained and unstained thin ground sections as well as microradiographed thick sections were produced for light microscopic morphometry and fluoroscopy. After 6 weeks, osteoclastic/osteoblastic activity was primarily observed in the graft-recipient contact area and in the intracortical compartment of the graft bone. New bone formation observed on the implant surface originated from the recipient site. The bone formation was evident also in the implant-graft interface. At 6 weeks the average bone fill of the implant threads was 28.4% which increased to 36.4% after 6 months as measured by morphometry. An average of 17.6% bony contact was measured after 6 weeks which increased to 29.7% 6 months after surgery. The graft bone had reduced in size from an average of 39.5% after 6 weeks down to 24.8% after 6 months (P \lt 0.05). It is concluded that the described experimental model can serve as a useful method for the study of implant healing in onlay grafts.

Journal Article↗
PubMed1998

Bovine hydroxyapatite for maxillary sinus grafting: comparative histomorphometric findings in sheep.

This experimental study in animals examines the value of bovine hydroxyapatite as grafting material in one-stage sinus lift procedure. The Schneiderian membrane was elevated from extraorally in 54 sinuses of 27 adult female mountain sheep. Two titanium plasma-flame-spray coated cylindrical implants were placed in each sinus in the same session. In 2 groups of 18 sinuses each, the subantral hollow space was filled with bovine hydroxyapatite and autogenous cancellous bone from the iliac crest, respectively. Eighteen sinuses that were left empty served as a control group. Polyfluorochrome sequential labelling (tetracycline, calcein green, alizarin complexon) was carried out 4 weeks postoperatively, 2 weeks before the animals were sacrificed, and halfway through the observation period to assess new bone formation. The selected observation periods were 12 weeks, 16 weeks, and 26 weeks. Six sinuses per observation period and test group were available for histologic evaluation. All implants showed favourable osseointegration into local bone. New bone formation was observed in a triangular area bound by the implant surface, the local buccal antral wall, and submucous connective tissue in all implants. Bovine hydroxyapatite adjacent to local bone was mainly surrounded by bone tissue. There was a great variation in the extent of bony sheathing of the hydroxyapatite particles, irrespective of the animals' survival time. Macrophages were found around HA particles that were not surrounded by bone as well as on free implant surfaces. The mean length of the bone-implant bond was 3.86 mm in the group of empty sinuses, 4.87 mm in the group augmented with hydroxyapatite, and 5.66 mm in the group augmented with autogenous cancellous bone. During the observation period, the relative length of bone in direct contact with the implant surface increased from 20.0% to 25.1% in the control group, from 30.4% to 35.5% in the group treated with autogenous cancellous bone, and from 27.4% to 34.7% in the HA group. Autogenous cancellous bone showed a significantly greater bone-implant contact than did the control group (P = 0.0004) and the HA group (P = 0.0499). When the apical implant portions were examined separately, both HA (P = 0.006) and autogenous cancellous bone (P = 0.005) showed a significantly better bone-implant bond than did the control group.

Animals↗
PubMed2003

Enzymatic method for assaying calcium in serum with Ca++ -ATPase.

A kinetic assay for total calcium in serum was developed which is based on the activation of Ca(++)-ATPase by free Ca(++) [Ca(++)](f) maintained by EGTA in the reaction mixture. The concentration of Ca(++)(f) was dependent on total reference calcium added or serum calcium. Ca(++)-ATPase activity was coupled to the reduction of NADH by pyruvate kinase (PK) and lactate dehydrogenase (LDH) and monitored by change in absorbance at 340 nm. The calcium in normal serum was 10.08 +/- 0.24 mg/dl (n = 35) by our method while with o-cresolphthalein complexone (CPC) method, the total calcium in the same 35 serum samples was 10.14 +/- 0.54 mg/dl. The range of within-run coefficient of variations (CVs) by this method was 0.9-2.87% at 8-12 mg/dl and day-to-day CVs were 0.72-3.17%. The presence of other ions and standard clinical interfering agents did not affect this assay system. The correlation between values obtained with our method (y) and CPC method (x) for normal serum was: y = 1.064x-0.580 mg/dl (r = 0.912, n = 59).

Adenosine Triphosphatases↗
PubMed2003

In vitro cariogenic potential of Candida albicans.

The adherence and dissociation of Candida albicans, C. tropicalis, Streptococcus mutans and S. sanguis to six substrates including hydroxylapatite (HAP) which exhibit various hydrophobicity, was examined by the use of a bioluminescent adenosine triphosphate (ATP) assay. Dissolution of HAP by C. albicans or S. mutans was determined spectrophotometrically by the use of o-cresolphthalein complexone. In the adherence of C. tropicalis, S. mutans and S. sanguis, the amount of adherent cells correlated with the hydrophobicity of the substrates. In contrast, the adherence of C. albicans to HAP was extraordinary high, although the adherence of the fungi also correlated with the hydrophobicity of the substrates, except for HAP. The yeasts attached to HAP was effectively removed by high concentration of either phosphate or calcium ions. The amount of calcium-release from HAP caused by C. albicans and S. mutans was 113 microg ml(-1) (final pH = 3.45), and 5.4 microg ml(-1) (final pH 4.81), respectively and the maximum growth of C. albicans and S. mutans was 10(7) cfu ml(-1) and 7.4 x 10(12) cfu ml(-1), respectively. The results, taken together, suggest that C. albicans adhere to HAP specifically through electrostatic interaction, and that, in a much smaller number (1.0/7.4 x 10(5)), C. albicans possesses the ability to dissolve HAP to a greater extent (approximately 20-fold) when compared with S. mutans.

Bacterial Adhesion↗
PubMed2003

Detecting microdamage in bone.

Fatigue-induced microdamage in bone contributes to stress and fragility fractures and acts as a stimulus for bone remodelling. Detecting such microdamage is difficult as pre-existing microdamage sustained in vivo must be differentiated from artefactual damage incurred during specimen preparation. This was addressed by bulk staining specimens in alcohol-soluble basic fuchsin dye, but cutting and grinding them in an aqueous medium. Nonetheless, some artefactual cracks are partially stained and careful observation under transmitted light, or epifluorescence microscopy, is required. Fuchsin lodges in cracks, but is not site-specific. Cracks are discontinuities in the calcium-rich bone matrix and chelating agents, which bind calcium, can selectively label them. Oxytetracycline, alizarin complexone, calcein, calcein blue and xylenol orange all selectively bind microcracks and, as they fluoresce at different wavelengths and colours, can be used in sequence to label microcrack growth. New agents that only fluoresce when involved in a chelate are currently being developed--fluorescent photoinduced electron transfer (PET) sensors. Such agents enable microdamage to be quantified and crack growth to be measured and are useful histological tools in providing data for modelling the material behaviour of bone. However, a non-invasive method is needed to measure microdamage in patients. Micro-CT is being studied and initial work with iodine dyes linked to a chelating group has shown some promise. In the long term, it is hoped that repeated measurements can be made at critical sites and microdamage accumulation monitored. Quantification of microdamage, together with bone mass measurements, will help in predicting and preventing bone fracture failure in patients with osteoporosis.

Animals↗
PubMedDate not supplied

[Results of fluorescence microscopy studies of bone healing by direct stimulation with bipolar impulse currents and with the interference current procedure in the animal experiment].

42 cross-breed rabbit bastards of either sex were osteotomized on the left proximal third of the tibia. A teflonisolated stable plating was made by means of the polychromatically KF-AO-instrumentarium. The animals were fluorescentlabelled in weekly intervals. Tetraverinex, alizarin complexon, fluorexon, xylenol orange and calceine were used as colours. The animals were stimulated in the bipolar squaretopped pulse current procedure (1 Hz and 10 Hz, resp., +/- 25 and +/- 50 microA, resp., intensity, permanent stimulation) or in the interference current procedure (oscillation frequency 100 Hz, intensity 1 mA, 4 hours daily). An osteotomized group served as a control. The undecalcified bone sections were quantitatively measured in the area of the periosteal and endoosteal accummulation seams as well as in the area of the Haversians canals and compared by means of multiple variance analyses. A delay in the Haversian remodelling within the first 2 weeks was found in the animals only osteotomized. This delay could not be detected in all electrically stimulated groups. The electrical stimulation leads to a shortening of the fracture healing period by skipping the physiologically occurring delay of the Haversian remodelling in fractures and osteotomies. Further on there was derived a growth function of the osteones as a regression function r (t) = a + beta X e gamma t. For the rabbit the concrete formula expression r (t) = 50.9 X e-0.094 X t + 17.4 for the animals not treated and r (t) = 42.9 X e-0.067 X t + 8.5 for the electrical stimulated animals has been found.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

Calcium levels in maternal milk: relationships with calcium intake during the third trimester of pregnancy.

The aim of the present study was to investigate the relationship of Ca intake and serum Ca levels during the third trimester of pregnancy with levels of the same mineral in transition milk (days 13-14 of lactation) and mature milk (day 40 of lactation). The study subjects were a group of fifty-seven healthy, lactating mothers aged between 18 and 35 years (mean 27 (SD 3.7) years) whose pregnancies and labour were attended by the Department of Obstetrics and Gynaecology of Cuenca INSALUD Hospital, Spain. Ca intake during the third trimester was determined by recording the consumption of foods over a 5 d period and by registering Ca provided by dietary supplements. The same method was used to investigate the intake of protein, vitamin D, fibre and Fe, nutrients that could affect the use of dietary Ca. Ca levels in maternal serum during this stage of pregnancy, during lactation and in transition and mature milk samples, were determined using 2-cresolphthalein complexone. During pregnancy 70.2% of subjects showed Ca intakes below 1100 mg/d (75th percentile). The consumption of Ca supplements was very small and hardly modified the mean quantity supplied by the diet. Subjects with an intake < 1100 mg/d showed no fall in Ca levels in serum, either during pregnancy or lactation, nor were decreased levels found in transition milk. However, these subjects showed lower Ca levels in mature milk (5.95 (SD 1.56) mmol/l) than did subjects with greater Ca intakes (6.82 (SD 1.31) mmol/l). This may suggest that breast-fed babies of mothers with lower Ca intakes during pregnancy also receive less Ca.

Adolescent↗
PubMed1976

Microdetermination of serum calcium by parallel fast analyzer.

An adaptation of a direct microdetermination of serum calcium based upon its colored reaction product with cresolphthalein complexone for use with a parallel fast analyzer is described. It is simple, reliable and clinically accurate. Twenty microliters of sample are employed in a final volume of 650 mul. Comparison studies with an atomic absorption method resulted in a correlation coefficinet (r) of 0.94. Replicate analyses at normal (8.5 mg. per dl.) and abnormal (12.1 mg. per dl) levels showed inter-day coefficients of variation of 3.2 and 2.3%, respectively. Assays of sera of male and female donors (age range: 17--60 years) showed normal ranges of 8.5--10.8 and 8.5--10.5 mg. per dl., respectively. Moderate hemolysis and jaundice and mild lipemia are associated with spuriously high results. Magnesium in serum showed no significant effect below 4.0 mEq. per l.

Adolescent↗
PubMed2000

Fish inner ear otoliths stop calcium incorporation after vestibular nerve transection.

Previous investigations revealed that the growth of fish inner ear otoliths (otolith size and calcium incorporation) depends on the amplitude and the direction of gravity, suggesting the existence of a (negative) feedback mechanism. In a search for the regulating unit, the vestibular nerve was unilaterally transected in neonatal swordtail fish (Xiphophorus helleri) which were subsequently incubated in the calcium-tracer alizarin-complexone. Calcium incorporation and thus otolith growth ceased on the operated head sides, indicating that the brain is significantly involved in regulating otolith growth.

Animals↗
PubMed1991

The healing of facial bone fractures by the process of secondary union.

The mechanism of healing of facial bone fractures was investigated in a rabbit model. Twelve New Zealand white rabbits underwent surgically induced fractures of the right infraorbital rim and fracture ostectomies (4 to 5 mm) of the left infraorbital rim. Animals were sacrificed 2, 4, and 8 weeks postfracture. Bone, including periosteum, obtained from each fracture or fracture osteoctomy site was divided longitudinally for hematoxylin and eosin staining, fluorescent microscopy, microangiography, and microradiography. Sequential fluorochrome labels of oxytetracycline (30 mg/kg), alizarin complexone (30 mg/kg), DCAF (20 mg/kg), and xylenol orange (90 mg/kg) were administered 24 hours preoperatively and at 1, 2, 4, and 8 weeks postfracture. All fracture and fracture ostectomy sites demonstrated vascular ingrowth, mineralization, and woven bone formation by 2 to 4 weeks postoperatively, beginning with a cartilage precursor. Subsequently, the woven bone was replaced with remodeled lamellar bone, resulting in complete bony healing by 8 weeks postoperatively. These steps were substantiated by microscopic, microradiographic, and radiologic examination of the specimens. This study demonstrates that fractures of the facial bones in a rabbit model heal by a process of new bone formation that resembles secondary union in endochondral bones.

Animals↗