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1.
Hum Reprod ; 36(7): 1907-1921, 2021 06 18.
Article in English | MEDLINE | ID: mdl-34052851

ABSTRACT

STUDY QUESTION: What is the transcriptome signature associated with poor performance of rescue IVM (rIVM) oocytes and how can we rejuvenate them? SUMMARY ANSWER: The GATA-1/CREB1/WNT signalling axis was repressed in rIVM oocytes, particularly those of poor quality; restoration of this axis may produce more usable rIVM oocytes. WHAT IS KNOWN ALREADY: rIVM aims to produce mature oocytes (MII) for IVF through IVM of immature oocytes collected from stimulated ovaries. It is not popular due to limited success rate in infertility treatment. Genetic aberrations, cellular stress and the absence of cumulus cell support in oocytes could account for the failure of rIVM. STUDY DESIGN, SIZE, DURATION: We applied single-cell RNA sequencing (scRNA-seq) to capture the transcriptomes of human in vivo oocytes (IVO) (n = 10) from 7 donors and rIVM oocytes (n = 10) from 10 donors. The effects of maternal age and ovarian responses on rIVM oocyte transcriptomes were also studied. In parallel, we studied the effect of gallic acid on the maturation rate of mouse oocytes cultured in IVM medium with (n = 84) and without (n = 85) gallic acid. PARTICIPANTS/MATERIALS, SETTING, METHODS: Human oocytes were collected from donors aged 28-41 years with a body mass index of <30. RNA extraction, cDNA generation, library construction and sequencing were performed in one preparation. scRNA-seq data were then processed and analysed. Selected genes in the rIVM versus IVO comparison were validated by quantitative real-time PCR. For the gallic acid study, we collected immature oocytes from 5-month-old mice and studied the effect of 10-µM gallic acid on their maturation rate. MAIN RESULTS AND THE ROLE OF CHANCE: The transcriptome profiles of rIVM/IVO oocytes showed distinctive differences. A total of 1559 differentially expressed genes (DEGs, genes with at least 2-fold change and adjusted P < 0.05) were found to be enriched in metabolic processes, biosynthesis and oxidative phosphorylation. Among these DEGs, we identified a repression of WNT/ß-catenin signalling in rIVM when compared with IVO oocytes. We found that oestradiol levels exhibited a significant age-independent correlation with the IVO mature oocyte ratio (MII ratio) for each donor. rIVM oocytes from women with a high MII ratio were found to have over-represented cellular processes such as anti-apoptosis. To further identify targets that contribute to the poor clinical outcomes of rIVM, we compared oocytes collected from young donors with a high MII ratio with oocytes from donors of advanced maternal age and lower MII ratio, and revealed that CREB1 is an important regulator. Thus, our study identified that GATA-1/CREB1/WNT signalling was repressed in both rIVM oocytes versus IVO oocytes and in rIVM oocytes of lower versus higher quality. Consequently we investigated gallic acid, as a potential antioxidant substrate in human rIVM medium, and found that it increased the mouse oocyte maturation rate by 31.1%. LARGE SCALE DATA: Raw data from this study can be accessed through GSE158539. LIMITATIONS, REASONS FOR CAUTION: In the rIVM oocytes of the high- and low-quality comparison, the number of samples was limited after data filtering with stringent selection criteria. For the oocyte stage identification, we were unable to predict the presence of oocyte spindle, so polar body extrusion was the only indicator. WIDER IMPLICATIONS OF THE FINDINGS: This study showed that GATA-1/CREB1/WNT signalling was repressed in rIVM oocytes compared with IVO oocytes and was further downregulated in low-quality rIVM oocytes, providing us the foundation of subsequent follow-up research on human oocytes and raising safety concerns about the clinical use of rescued oocytes. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the Collaborative Research Fund, Research Grants Council, C4054-16G, and Research Committee Funding (Research Sustainability of Major RGC Funding Schemes), The Chinese University of Hong Kong. The authors have no conflicts of interest to declare.


Subject(s)
Oocytes , Ovulation Induction , Animals , Cumulus Cells , Female , In Vitro Oocyte Maturation Techniques , Mice , Oogenesis , Sequence Analysis, RNA
2.
Osteoporos Int ; 31(8): 1507-1516, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32246167

ABSTRACT

The receptor activator of nuclear factor-kappa B ligand (RANKL)/RANK/osteoprotegerin system is dysregulated in hyperparathyroid bone diseases. The introduction of denosumab preceding elective surgery as an alternative option when surgery is not possible immediately. INTRODUCTION: The effects of denosumab on vascular calcification in patients with chronic renal failure and low bone mass have been a subject of interest. Therefore, this investigation aimed to determine the short-term changes in vascular calcification after denosumab treatment using a serial electrocardiography-gated computed tomography (CT) to measure coronary artery calcification (CAC) in patients with secondary hyperparathyroidism (SHPT) and low bone mass. METHODS: This 6-month study enrolled patients with SHPT and low bone mass (T-score < - 2.5) owing to dialysis. The 2 groups administered denosumab at a dose of 60 mg (denosumab group), and conventional treatment (control group) had 21 patients each. All patients underwent CT scans at baseline and at the follow-up examination at 6 months to determine the bone mineral density and CAC. RESULTS: The control group demonstrated a significant increase in Agatston scores (187.79 ± 72.27) (P = 0.004). However, no significant change was noted in the denosumab group (P = 0.41). In the denosumab group, only the baseline serum alkaline phosphatase levels correlated negatively with changes in the CAC score (P = 0.01); the baseline alkaline phosphatase levels were the deciding biomarkers for non-responsive CAC scores by Berry Criteria after denosumab treatment (P = 0.02). The denosumab group demonstrated significantly increased bone mineral density in the femoral neck and lumbar spine (P < 0.01). CONCLUSION: The findings provide evidence that denosumab may suppress the progression of CAC and also regress osseous calcification in severe cases of high bone turnover.


Subject(s)
Bone Density Conservation Agents , Calcinosis , Cardiovascular Diseases , Denosumab , Hyperparathyroidism, Secondary , Kidney Failure, Chronic , Bone Density , Bone Density Conservation Agents/therapeutic use , Calcinosis/prevention & control , Cardiovascular Diseases/prevention & control , Denosumab/therapeutic use , Humans , Hyperparathyroidism, Secondary/complications , Hyperparathyroidism, Secondary/drug therapy , Kidney Failure, Chronic/complications , Kidney Failure, Chronic/therapy , Pilot Projects , Renal Dialysis/adverse effects
3.
Osteoporos Int ; 30(2): 519-523, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30171299

ABSTRACT

Denosumab, a novel agent that inhibits osteoclasts, reduces the risk of fracture in patients with osteoporosis. However, worsening of hypophosphatemia and other symptoms may be induced by denosumab in patients with pre-existing hypophosphatemic osteomalacia. A 58-year-old man with hepatitis B presented with diffuse bone pain and muscle weakness. Denosumab was prescribed by the orthopedist according to documented low bone mass and spine compression fracture. After administering denosumab, the patient's bone pain worsened, and he later developed a right tibia stress fracture. His condition was diagnosed as adult-onset hypophosphatemic osteomalacia complicated by multiple bone fractures, which resulted from Fanconi syndrome with proximal tubulopathy due to tenofovir disoproxil fumarate (TDF) treatment for his hepatitis B. Denosumab use leads to aggressive hypophosphatemic osteomalacia and the complication of stress fractures, because of its effects on bone resorption. Physicians should be aware that in patients with chronic hepatitis B monoinfection who are administered TDF therapy, bone pain or fracture is possible but preventable by timely monitoring of serum phosphate levels. Denosumab should not be used in patients with untreated osteomalacia or vitamin D deficiency, as it may lead not only to hypocalcemia but also to hypophosphatemia in these patients.


Subject(s)
Bone Density Conservation Agents/adverse effects , Denosumab/adverse effects , Fanconi Syndrome/chemically induced , Hypophosphatemia/chemically induced , Osteomalacia/drug therapy , Antiviral Agents/adverse effects , Antiviral Agents/therapeutic use , Bone Density Conservation Agents/therapeutic use , Denosumab/therapeutic use , Fanconi Syndrome/complications , Fractures, Stress/diagnostic imaging , Fractures, Stress/etiology , Hepatitis B, Chronic/drug therapy , Humans , Male , Middle Aged , Osteomalacia/diagnostic imaging , Radiography , Radionuclide Imaging , Tenofovir/adverse effects , Tenofovir/therapeutic use , Tibial Fractures/diagnostic imaging , Tibial Fractures/etiology , Tomography, X-Ray Computed
4.
Int J Clin Pract ; 69(12): 1473-85, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26299643

ABSTRACT

BACKGROUND: An increased risk for ischaemic stroke has been reported in young hyperthyroidism patients independent of atrial fibrillation (AF). However, whether the use of antithyroid drugs in hyperthyroidism patients can reduce the occurrence of ischaemic stroke remains unclear. METHODS: A total of 36,510 newly diagnosed hyperthyroidism patients during 2003-2006 were identified from the Taiwan National Health Insurance Research database. Each patient was individually tracked for 5 years from their index date (beginning the antithyroid drugs) to identify those who suffered from new episode of ischaemic stroke. Medication possession ratio (MPR) was used to represent the antithyroid drug compliance. The association between the MPR and the risk of stroke was examined. RESULTS: The stroke incidence rates for hyperthyroidism patients with age < 45 years and age ≥ 45 years were 0.42 and 3.76 per 1000 person-years, respectively. The patients aged < 45 years with MPR < 0.2 (adjusted hazard ratio, HR, 2.30; 95% CI, 1.13-4.70; p = 0.02) and 0.2 ≤ MPR < 0.4 (adjusted HR, 2.24; 95% CI, 1.06-4.72; p = 0.035) had a significantly increased risk of ischaemic stroke as compared to those with ≥ 0.6. In patients of the age ≥ 45 years, only the patients with MPR < 0.2 (adjusted HR, 1.44; 95% CI, 1.03-2.01; p = 0.036) had a significantly higher risk of ischaemic stroke as compared to those with MPR ≥ 0.6. In hyperthyroidism patients without AF, good antithyroid drugs compliance also reduced the incidence of stroke significantly (adjusted HR, range: 1.52-1.61; p = 0.02); but not in hyperthyroidism with AF. CONCLUSION: Hyperthyroidism patients with good antithyroid drug compliance had a lower risk of ischaemic stroke than patients with poor compliance.


Subject(s)
Antithyroid Agents/therapeutic use , Hyperthyroidism/drug therapy , Medication Adherence , Stroke/epidemiology , Adult , Aged , Female , Humans , Hyperthyroidism/complications , Incidence , Male , Middle Aged , Proportional Hazards Models , Risk Factors , Stroke/etiology , Taiwan/epidemiology , Young Adult
5.
Eur J Neurol ; 20(8): 1191-7, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23581395

ABSTRACT

BACKGROUND AND PURPOSE: Elevated plasma total homocysteine level (tHcy) is associated with increased risk of dementia via increased white matter changes or reduction in cortical volume. Whether tHcy has an independent impact on regional perfusion and if it can predict a more rapid cognitive decline in mild Alzheimer dementia (AD) warrants investigation. METHODS: Eighty AD patients with a clinical dementia rating of 1 were enrolled. Their Cognitive Ability Screening Instrument (CASI) scores on enrolment and after 1 year of follow-up as well as their perfusion index (PI) from single photon emission computed tomography upon enrolment were analyzed. RESULTS: In cross-sectional analysis, elevated tHcy was associated with lower frontal PI independent of cerebrovascular risk factors (ß = -0.35, P = 0.009). The CASI scores correlated with temporo-parietal PI (Pearson r range 0.3-0.39, P < 0.01) but not with tHcy or frontal PI. By longitudinal analysis, only tHcy level was related to a more rapid cognitive decline (odds ratio for executive function score 1.82; odds ratio for total CASI score 1.74). CONCLUSIONS: Cognitive performance in mild AD can be reflected by hypo-perfusion of the temporo-parietal region while frontal hypo-perfusion may be mediated by tHcy. tHcy level is an independent risk factor for rapid cognitive decline, especially in the executive function.


Subject(s)
Alzheimer Disease/blood , Alzheimer Disease/psychology , Cerebrovascular Circulation/physiology , Cognition Disorders/blood , Cognition Disorders/psychology , Homocysteine/blood , Aged , Aged, 80 and over , Alzheimer Disease/complications , Brain/diagnostic imaging , Cognition Disorders/etiology , Disease Progression , Female , Humans , Male , Middle Aged , Neuropsychological Tests , Perfusion , Risk Factors , Tomography, Emission-Computed, Single-Photon
6.
Rev Sci Instrum ; 81(5): 054701, 2010 May.
Article in English | MEDLINE | ID: mdl-20515160

ABSTRACT

Mode converters are critical for frequency-tunable terahertz gyrotrons. This study reports the development of a broadband TE(02) mode converter centered at 0.2 THz. An octafeed sidewall coupling structure was employed and the mode purity was analyzed. The converter was built using the technique of x-ray microfabrication. The x rays irradiated on the SU-8 resist and generated a template of very high thickness of 1.295 mm. Pulse electroplating technique was used to deposit copper on the structure along the template. The parts then went through precise machining and the residual resist was removed via high-flux radical etching. A computer-aided diagnostic system was introduced to measure the performance of the converter. Results suggest that the frequency response of resistivity should be taken into consideration for the devices in terahertz region.

7.
Int J Immunopathol Pharmacol ; 22(2): 311-22, 2009.
Article in English | MEDLINE | ID: mdl-19505385

ABSTRACT

To determine the expression of components in Toll-like receptors (TLRs)/Nod-like receptors (NLRs)/inflammasome/caspase-1/interleukin (IL-1)-beta pathway, we examined the expression profiles of those genes by analyzing the data from expression sequence tag cDNA cloning and sequencing. We made several important findings: firstly, among 11 tissues examined, vascular tissues and heart express fewer types of TLRs and NLRs than immune and defense tissues including blood, lymph nodes, thymus and trachea; secondly, brain, lymph nodes and thymus do not express proinflammatory cytokines IL-1beta and IL-18 constitutively, suggesting that these two cytokines need to be upregulated in the tissues; and thirdly, based on the expression data of three characterized inflammasomes (NALP1, NALP3 and IPAF inflammasome), the examined tissues can be classified into three tiers: the first tier tissues including brain, placenta, blood and thymus express inflammasome(s) in constitutive status; the second tier tissues have inflammasome(s) in nearly-ready expression status (with the requirement of upregulation of one component); the third tier tissues, like heart and bone marrow, require upregulation of at least two components in order to assemble functional inflammasomes. Our original model of three-tier expression of inflammasomes would suggest a new concept of tissue inflammation privilege, and provides an insight to the differences among tissues in initiating acute inflammation in response to stimuli.


Subject(s)
Cardiovascular System/immunology , Inflammation/genetics , Signal Transduction/genetics , Adaptor Proteins, Signal Transducing/genetics , Apoptosis Regulatory Proteins/genetics , CARD Signaling Adaptor Proteins/genetics , Calcium-Binding Proteins/genetics , Carrier Proteins/genetics , Cells, Cultured , Endothelial Cells/immunology , Gene Expression Profiling , Gene Expression Regulation , Humans , Inflammation/immunology , Interleukin-18/genetics , Interleukin-1beta/genetics , NLR Family, Pyrin Domain-Containing 3 Protein , NLR Proteins , Nod Signaling Adaptor Proteins/genetics , Signal Transduction/immunology , Toll-Like Receptors/genetics , Tumor Necrosis Factor-alpha/metabolism
8.
J Fish Biol ; 74(2): 357-76, 2009 Feb.
Article in English | MEDLINE | ID: mdl-20735565

ABSTRACT

Morphometric and genetic data were used to compare two sympatric and morphologically similar species, Acanthopagrus berda and Acanthopagrus taiwanensis, in Dapeng Bay, South-western Taiwan. A principle component analysis of morphological data indicated a distinction between the two species, with pectoral fin length and eye diameter accounting for 32.27% of the variation. Interspecific sequence divergence, based on mtDNA cytochrome b (0.118 +/- 0.01), was larger than intraspecific divergences between haplotypes (0.007 for A. taiwanensis and 0.003 for A. berda). Individuals of the two species clustered into different groups in three phylogenetic trees with 100% bootstrap support. The mean observed heterozygosity for eight microsatellite loci was 0.471 +/- 0.202 for A. taiwanensis and 0.637 +/- 0.145 for A. berda. Nei's unbiased measure of interspecific genetic distance (D(S)) was 1.334. F(ST) (0.134) and R(ST) (0.404) values indicated significant differentiation between species. An unrooted neighbour-joining tree was constructed by allele-sharing distances and the factorial correspondence analysis split all specimens into two distinct clusters. The results of morphometric, mtDNA and microsatellite analyses indicated the presence of two species, A. taiwanensis and A. berda.


Subject(s)
Genetic Variation , Perciformes/classification , Phylogeny , Animals , Cytochromes b/genetics , DNA, Mitochondrial/genetics , Haplotypes , Microsatellite Repeats , Perciformes/anatomy & histology , Perciformes/genetics , Sequence Analysis, DNA , Taiwan
9.
Opt Express ; 16(25): 20759-73, 2008 Dec 08.
Article in English | MEDLINE | ID: mdl-19065215

ABSTRACT

The edge emission from AlGaInP light-emitting diodes showed a red-shifted peak in addition to the peak of surface emission. This shift resulted from the quantum-well absorption of the guided wave. Although the shift degrades the color quality and the extraction efficiency of the device, it helps elucidate many important optical properties of the material and the dynamics of carrier recombination, including the electron-hole recombination lifetime, the optical joint density of state, the spontaneous emission spectrum and the absorption spectrum. A simple concept of the bimolecular recombination is established. The corresponding coefficient can be expressed by a simple formula and was therefore determined.


Subject(s)
Computer-Aided Design , Lighting/instrumentation , Models, Theoretical , Quantum Dots , Aluminum Compounds/chemistry , Computer Simulation , Electrons , Equipment Design , Equipment Failure Analysis , Light , Scattering, Radiation , Semiconductors
10.
Environ Sci Technol ; 42(13): 5008-13, 2008 Jul 01.
Article in English | MEDLINE | ID: mdl-18678041

ABSTRACT

Two approaches were undertaken to characterize the arsenic (As) content of Chinese rice. First, a national market basket survey (n = 240) was conducted in provincial capitals, sourcing grain from China's premier rice production areas. Second, to reflect rural diets, paddy rice (n = 195) directly from farmers fields were collected from three regions in Hunan, a key rice producing province located in southern China. Two of the sites were within mining and smeltery districts, and the third was devoid of large-scale metal processing industries. Arsenic levels were determined in all the samples while a subset (n = 33) were characterized for As species, using a new simple and rapid extraction method suitable for use with Hamilton PRP-X100 anion exchange columns and HPLC-ICP-MS. The vast majority (85%) of the market rice grains possessed total As levels < 150 ng g(-1). The rice collected from mine-impacted regions, however, were found to be highly enriched in As, reaching concentrations of up to 624 ng g(-1). Inorganic As (As(i)) was the predominant species detected in all of the speciated grain, with As(i) levels in some samples exceeding 300 ng g(-1). The As(i) concentration in polished and unpolished Chinese rice was successfully predicted from total As levels. The mean baseline concentrations for As(i) in Chinese market rice based on this survey were estimated to be 96 ng g(-1) while levels in mine-impacted areas were higher with ca. 50% of the rice in one region predicted to fail the national standard.


Subject(s)
Arsenic/analysis , Food Contamination/analysis , Mining , Oryza/chemistry , China , Chromatography, High Pressure Liquid , Chromatography, Ion Exchange , Mass Spectrometry
11.
Phys Rev E Stat Nonlin Soft Matter Phys ; 74(1 Pt 2): 016402, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16907193

ABSTRACT

The gyrotron traveling-wave amplifier employing the distributed-loss scheme is capable of very high gain and effective in suppressing the global absolute instabilities. This study systematically characterizes the local absolute instabilities and their transitional behavior. The local absolute instabilities are analyzed using a model that incorporates the penetration of the field from the copper section into the lossy section. The axial modes were characterized from the perspective of beam-wave interaction and were found to share many characteristics with the global modes. The transition from global modes to local modes as the distributed loss increases was demonstrated. The electron transit angle in the copper section, which determines the feedback criterion, governs the survivability of an oscillation. In addition, the oscillation thresholds predicted using this model are more accurate than those obtained using a simplified model.

12.
Phys Rev E Stat Nonlin Soft Matter Phys ; 70(5 Pt 2): 056402, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15600760

ABSTRACT

The absolute instability is a subject of considerable physics interest as well as a major source of self-oscillations in the gyrotron traveling-wave amplifier (gyro-TWT). We present a theoretical study of the absolute instabilities in a TE01 mode, fundamental cyclotron harmonic gyro-TWT with distributed wall losses. In this high-order-mode circuit, absolute instabilities arise in a variety of ways, including overdrive of the operating mode, fundamental cyclotron harmonic interactions with lower-order modes, and second cyclotron harmonic interaction with a higher-order mode. The distributed losses, on the other hand, provide an effective means for their stabilization. The combined configuration thus allows a rich display of absolute instability behavior together with the demonstration of its control. We begin with a study of the field profiles of absolute instabilities, which exhibit a range of characteristics depending in large measure upon the sign and magnitude of the synchronous value of the propagation constant. These profiles in turn explain the sensitivity of oscillation thresholds to the beam and circuit parameters. A general recipe for oscillation stabilization has resulted from these studies and its significance to the current TE01 -mode, 94-GHz gyro-TWT experiment at UC Davis is discussed.

13.
Spine (Phila Pa 1976) ; 26(22): 2478-84, 2001 Nov 15.
Article in English | MEDLINE | ID: mdl-11707714

ABSTRACT

STUDY DESIGN: An anatomic study of cervical vertebral bodies. OBJECTIVES: To provide quantitative information on the cortical shell architecture of the middle and lower cervical vertebral bodies. SUMMARY OF BACKGROUND DATA: Some external dimensions have been measured, but little quantitative data exists for the cortical shell architecture of the vertebral bodies of the cervical spine. METHODS: Twenty-one human cervical vertebral bodies (C3-C7) were sectioned along parasagittal planes into five 1.7-mm thin slices for each vertebra. Radiographs of each slice were digitized, and external and internal dimensions were measured. Averages and standard deviations were computed. Single factor analysis of variance was used to determine significant (P < 0.05) differences between the vertebral levels. RESULTS: The superior endplate was thickest in the posterior region (range 0.74-0.89 mm) and thinnest in the anterior region (range 0.44-0.56 mm). The inferior endplate was thickest in the anterior region (range 0.61-0.81 mm) and thinnest in the posterior region (range 0.49-0.62 mm). In the central region, the superior endplate (range 0.42-0.58 mm) was thinner than the inferior endplate (range 0.53-0.64 mm). Variation with vertebral level was dependent on the dimension studied. CONCLUSIONS: Comprehensive quantitative anatomic data of the middle and lower cervical vertebral bodies have been obtained. This may be useful in improving the understanding of the three-column and other vertebral-fracture theories, the fidelity of the finite element models of cervical spine, and the designs of surgical instrumentation.


Subject(s)
Cervical Vertebrae/anatomy & histology , Cadaver , Cervical Vertebrae/diagnostic imaging , Electronic Data Processing , Humans , Radiography
15.
Sheng Wu Gong Cheng Xue Bao ; 17(6): 678-82, 2001 Nov.
Article in Chinese | MEDLINE | ID: mdl-11910764

ABSTRACT

To analyze the bone-inducing activity of C terminal of hBMP-2 and get a new recombinant product of hBMP-2, the gene encoding 102 aa of hBMP-2 mature peptide C terminal was cloned and expressed in E. coli and the first Cys was mutated with Ser. The fragments encoding the target peptide were amplified and cloned into heat-inducible expression vector pDH and transformed into E. coli DH5 alpha. After induction, a new protein bond appeared on the SDS-PAGE. The expressed products amounted to 30% of the total bacterial protein, which existed in the form of inclusion body. The products of bacterial lysates were purified through the ion-exchange chromatography. The denatured proteins were dialysed and diluted directly into the refolding buffer. The renatured products were implanted into mouse thigh muscles to analyze their bone-inducing activity respectively. The results of histological assay showed that the 102 peptide of hBMP-2 could ectopically induce formation of bone, while the mutated 102 peptide of hBMP-2 could not. It suggested hBMP-2 102 peptide still had bone-inducing activity. The first Cys of hBMP-2 mature peptide might be necessary for integrity of three pairs of disulfide bond, and also essential for bone-inducing activity of hBMP-2.


Subject(s)
Bone Morphogenetic Proteins/pharmacology , Osteogenesis/drug effects , Peptide Fragments/pharmacology , Transforming Growth Factor beta , Animals , Biological Assay , Bone Morphogenetic Protein 2 , Bone Morphogenetic Proteins/biosynthesis , Bone Morphogenetic Proteins/genetics , Cysteine/genetics , Escherichia coli/genetics , Femur/growth & development , Humans , Mice , Osteogenesis/physiology , Peptide Fragments/biosynthesis , Peptide Fragments/genetics , Point Mutation , Protein Engineering/methods , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , Recombinant Proteins/pharmacology
16.
Spine (Phila Pa 1976) ; 25(10): 1197-205, 2000 May 15.
Article in English | MEDLINE | ID: mdl-10806495

ABSTRACT

STUDY DESIGN: The internal architecture of cervical spine pedicles was investigated by thin sectioning and digitization of radiographic images. OBJECTIVES: To provide quantitative information on the internal dimensions and cortical shell thicknesses of the middle and lower cervical pedicles. SUMMARY OF BACKGROUND DATA: Although there have been a number of studies presenting data on the external dimensions of the cervical pedicle, little is known regarding its internal architecture and cortical shell thickness along the pedicle axis. METHODS: Twenty-five human cervical vertebrae (C3-C7) were secured to a thin-sectioning machine to produce three 0.7-mm-thick pedicle slices along its axis. Plain radiographs of the pedicle slices were scanned and digitized to facilitate measurement of the internal dimensions. Computer software was specifically developed to determine the external dimensions (i.e., pedicle height and width) and the internal dimensions (i.e., cortical shell thicknesses of the superior, inferior, lateral, and medial walls and the cancellous core height and width) of cervical pedicles. RESULTS: Superior and inferior wall cortical thicknesses of pedicle thin slices were similar, whereas the lateral wall cortical thickness was significantly smaller than the medial wall thickness. The medial cortical shell (average value range: 1.2-2.0 mm) was measured to be 1.4 to 3.6 times as thick as the lateral cortical shell (average value range: 0.4-1.1 mm). When medial and lateral cortical thicknesses were normalized for external dimensions, the combined cortical shell thickness was thinnest at C7 (average value range: 18. 6-25.6% of the external width), and this result was statistically significant when compared with other vertebral levels. CONCLUSIONS: The cervical pedicle is a complex, three-dimensional structure exhibiting extensive variability in internal morphology. Characteristics of the cervical pedicle at different spinal levels must be noted before transpedicular screw fixation.


Subject(s)
Cervical Vertebrae/anatomy & histology , Bone Screws , Cervical Vertebrae/diagnostic imaging , Cervical Vertebrae/surgery , Humans , Joint Instability/surgery , Microtomy , Radiography , Reference Standards , Spinal Fusion
17.
Eur Spine J ; 9(1): 61-6, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10766079

ABSTRACT

Transpedicular screw fixation has recently been shown to be successful in stabilizing the middle and lower cervical spine. Controversy exists, however, over its efficacy, due to the smaller size of cervical pedicles and the proximity of significant neurovascular structures to both lateral and medial cortical walls. To aid the spinal surgeon in the insertion of pedicle screws, a number of studies have been performed to quantify the gross dimensions and angulations of the cervical pedicle. Notwithstanding these quantitative studies, there has been a conspicuous absence of research reporting the qualitative characteristics of the cervical pedicle. The purpose of our study was to provide comparative graphical data that would systematically document the anatomic variability in cervical pedicle morphology. Such information should better elucidate the complexity of the pedicle as a three-dimensional structure and provide the spinal surgeon with a more complete understanding of cervical pedicle architecture. Twenty-six human cervical vertebrae (C3-C7) from six fresh-frozen spines were secured to a thin sectioning apparatus to produce three 0.7-mm-thick pedicle slices along its axis. Radiographs taken of these pedicle slices were scanned, digitized, and traced to facilitate visual comparison. The pedicle slices were found to exhibit substantial variability in composition and shape, not only between individual spines and vertebral levels, but also within the pedicle axis. However, the lateral cortex was consistently found to be thinner than the medial cortex in all samples. These physical findings must be noted by surgeons attempting transpedicular screw fixation in the cervical spine.


Subject(s)
Bone Screws , Cervical Vertebrae/anatomy & histology , Spinal Fusion/instrumentation , Cervical Vertebrae/surgery , Humans , In Vitro Techniques , Spinal Diseases/surgery
18.
Gene ; 211(1): 49-55, 1998 Apr 28.
Article in English | MEDLINE | ID: mdl-9573337

ABSTRACT

Isogenes are highly homologous to each other and are often difficult to ascertain, as has been the case with metallothionein, a metal-binding protein rich in cysteines. Conventional separation of metallothionein isoforms relied on ion exchange chromatography of the proteins, or screening for the sequences from gene libraries. In this study, a combination of RT-PCR and partial protein sequencing is used in the identification of metallothionein isogenes expressed in porcine liver. By this approach, we have identified expressed coding sequences which constitute 10 new isogenes. Of the four known groups of metallothioneins (MT), phylogenetic analyses place these pig isogenes in the MT-1 group, except two which are identified as being closely related to MT-2, and none in groups 3 and 4. The isogenes are thus namedpMT-1a to -1g, andpMT-2a and -2b. While each of the isogene sequences is unique, two isogenes,pMT-1e1 andpMT-1e2, share an identical amino acid sequence, differing only in specific codons. Two others,pMT-1b andpMT-1g, have a cysteine substituted by arginine, the first such sequence ever detected in MT.pMT-2a andpMT-2b are closely aligned with the MT-2 group of vertebrates, in spite of the absence of a characteristic acidic amino acid at position 10 or 11, common in other mammalian metallothioneins.


Subject(s)
Liver/chemistry , Metalloproteins/chemistry , Metallothionein/chemistry , Amino Acid Sequence , Animals , Cloning, Molecular , Molecular Sequence Data , Peptide Fragments/chemistry , Phylogeny , Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis , Sequence Homology, Nucleic Acid , Swine
19.
Invest Radiol ; 28(2): 142-5, 1993 Feb.
Article in English | MEDLINE | ID: mdl-8444570

ABSTRACT

RATIONALE AND OBJECTIVES: At many institutions, three-dimensional magnetic resonance imaging (MRI) is routinely used for examination of the knee. Multiplanar reconstruction (MPR) is a method of displaying three-dimensional datasets. The authors assessed the usefulness of MPR for evaluating knee MRI datasets by comparing readers' performance using MPR and conventional film MRIs. METHODS: Eight patients with internal derangement of the knee were studied. All had MRI datasets acquired in the sagittal plane using a three-dimensional gradient-echo fast imaging with steady-state precession (FISP) sequence. Arthroscopic surgery after MRI confirmed the presence of 6 anterior cruciate ligament (ACL) tears, 11 meniscal tears, and 5 normal menisci in this group. Four blinded readers, experienced in MRI of the knee, evaluated the images. The MRI datasets were then loaded onto a three-dimensional workstation and interpreted by the same readers using MPR. The MRI findings were correlated with arthroscopy. RESULTS: For diagnosis of tears of the ACL, sensitivity was 96% and specificity was 100% for both films and MPR. For detecting meniscal tears, sensitivity was 55% and specificity was 90%, using the filmed images, versus 64% and 85%, respectively, with MPR. These differences were not statistically significant by the sign test. Total time (technologist processing time plus radiologist reading time) for MPR was greater than for film interpretation by a factor of 1.12 (P < .05), and radiologist reading time for MPR was greater by a factor of 1.88 (P < .05). CONCLUSIONS: For sagittal three-dimensional FISP MRI datasets, real-time MPR is comparable with film interpretation for evaluation of ACL and meniscal injuries, but it increases the time required for diagnosis.


Subject(s)
Image Processing, Computer-Assisted , Knee Injuries/diagnosis , Magnetic Resonance Imaging/methods , Adolescent , Adult , Female , Humans , Ligaments, Articular/injuries , Ligaments, Articular/pathology , Male , Menisci, Tibial/pathology , Middle Aged , Tibial Meniscus Injuries
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