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Biomedical subjects

H Ding

Publications and source records attributed to H Ding.

At least 91 records · Page 5Linked to original sources
PubMed1999

Elevated myocardial cytosolic calcium impairs insulin-like growth factor-1-stimulated protein synthesis in chronic renal failure.

Rats and humans with chronic renal failure (CRF) are reported to have resistance to recombinant human insulin-like growth factor-1 (rhIGF-1). Because basal cytosolic calcium ([Ca2+]i), a second messenger, may be increased in CRF, this study was conducted to examine whether elevated basal [Ca2+]i may cause resistance to IGF-1. Cardiomyocytes from four groups of rats were studied: untreated CRF, CRF with parathyroidectomy (PTX), CRF with the calcium channel blocker felodipine (F), and sham operation of the kidney (SO). CRF was created by ligation of two-thirds of the left renal artery and contralateral nephrectomy. Rats from each group were pair-fed the same diet for 20 to 22 d. Basal [Ca2+]i in cardiomyocytes (nM) in the CRF rats (102.0 +/- 2.8; SEM), was significantly higher than in each of the CRF-PTX, CRF-F, and SO groups (65.2 +/- 1.9, 63.8 +/- 2.6, and 63.5 +/- 2.0, respectively; P < 0.01). rhIGF-1 increased cardiomyocyte [Ca2+]i in all four groups of rats. The rise in [Ca2+]i was significantly diminished in the CRF rats (P < 0.05) and did not differ among the CRF-PTX, CRF-F, and SO rats. Protein synthesis after incubation with 0, 50, 100, 200, or 400 ng/ml rhIGF-1 was lower in cardiomyocytes from CRF rats than in each of the other three groups (P < 0.05) and was significantly less in the CRF-F rats compared with SO animals. IGF-1 receptor mRNA and IGF-1 receptor number and affinity were not different among the four groups. These findings suggest that cardiomyocytes from CRF rats display elevated basal [Ca2+]i and attenuated rhIGF-1-induced increase in [Ca2+]i; basal protein synthesis is decreased, and IGF-1-stimulated protein synthesis is impaired; elevated basal [Ca2+]i seems to contribute to this diminished response to rhIGF-1.

Animals↗
PubMed1999

Transcriptional regulation via redox-sensitive iron-sulphur centres in an oxidative stress response.

Genetic responses to oxidative stress are triggered by excessive levels of agents such as superoxide. The soxRS regulon of Escherichia coli includes at least a dozen oxidative-stress and antibiotic-resistance genes that are activated by the SoxS protein, the synthesis of which is controlled by the redox-sensing SoxR protein. SoxR is a homodimer of 17 kDa subunits, each of which contains a [2Fe-2S] cluster. Transcriptional activation by SoxR is controlled by the oxidation state of these metal centres. In the absence of oxidative stress, the [2Fe-2S] centres are in the reduced form and the protein is inactive, although it still binds the soxS promoter. Agents that generate superoxide in the cell (e.g. paraquat) cause rapid oxidation of the metal centres, which triggers the transcriptional activity of SoxR; removal of the oxidative stress is followed by rapid re-reduction of the [2Fe-2S] centres. This facile mechanism links oxidation state to control of protein activity and may be used widely to allow cells to respond to oxidative stress.

Allosteric Regulation↗
PubMed1999

[The relationship between polymorphism of LMP2 and LMP7 genes and the phenotype of ankylosing spondylitis].

OBJECTIVE: To evaluate the relationship of the polymorphism of LMP2 and LMP7 genes and the phenotype of patients with ankylosing spondylitis(AS). METHODS: One hundred and eighteen blood samples were collected from patients with AS and acute anterior uveitis(AAU). HLA-B(27) typing was performed and LMP2 and LMP7 amplified fragment length polymorphisms were determined after PCR and digestion with Cfo I restriction enzyme in the study. RESULTS: There was a significantly increased LMP2 BB genotype frequency in patients with AS+AAU(86.9%) and AAU(87%), compared with controls(53.3%) and AS patients(63.1%)(P<0.05); the odds ratio(OR) relating LMP2 BB and AAU was 5.83, and the OR for AS+AAU was 3.71. No difference was found between adult AS and junior AS and between AS patients with and without extraspinal arthritis. No difference was found in LMP7 genotype. CONCLUSION: The polymorphism of LMP2 gene is strongly associated with the development of AAU in AS patients and in uncomplicated AAU patients, but not with the age at onset and the type of joints affected.

Adolescent↗
PubMed1999

Demethylation capacity of human fetal adrenal mitochondrial cytochrome P-450 in vitro.

AIM: To explore the capacity and characteristics of adrenal mitochondria to metabolize xenobiotics in vitro in human fetus. METHODS: Subcellular fractions of fetal adrenal were prepared by differential centrifugation. Mitochondrial P-450 system was proved by spectral analyses and SDS-PAGE. The formaldehyde formation contents were measured with Nash reagent. RESULTS: The erythromycin N-demethylation linearly increased in the protein concentration (1-4 mg)- and incubation time (10-30 min)-dependent manners. A typical concentration-effect relationship appeared with erythromycin 0.067-1 mmol.L-1 and a positive correlation (r = 0.641, P < 0.05) existed between erythromycin N-demethylation and gestation months. The N-demethylation values (nmol.s-1/g protein) of erythromycin (2.7 +/- 0.8), benzfetamine (1.1 +/- 0.5), and aminophenazone (0.9 +/- 0.4) in mitochondria were 89% (P > 0.05), 162% (P < 0.01), and 62% (P < 0.01), respectively, of those in microsomes. There was correlation between mitochondria and microsomes in the N-demethylation of erythromycin (r = 0.708, P < 0.05) and benzfetamine (r = 0.707, P < 0.05). Troleandomycin stimulated erythromycin N-demethylation in adrenal mitochondria as well as in adrenal and liver microsomes in vitro. CONCLUSION: Fetal adrenal mitochondria, with multiple P-450 isoforms and greater capacity of demethylation, play a role in drug-metabolism during fetal development.

Adrenal Glands↗
PubMed1999

[Analysis of national maternal mortality rate surveillance. Maternal Death Investigation Cooperative Group of China].

OBJECTIVE: To analyze the results of maternal mortality rate (MMR) surveillance in 247 monitoring areas. METHODS: Based on the demographic characteristics of different areas, stratum sampling method was used, and 247 monitoring points from 30 provinces, autonomous regions and cities covered about 100 million population were selected in China in 1989-1995. RESULTS: In the 7 years of monitoring there were 8,709,220 cases of live births with 5,984 cases of maternal deaths in these monitoring points. The average MMR was 77.4/100,000 with 45.5/100,000 in urban areas and 95.4/100,000 in rural areas. There were significant differences of MMR from area to area from 26.0/100,000 to 308.0/100,000. The causes of maternal deaths in succession were obstetric hemorrhage, pregnancy induced hypertension, heart diseases, amniotic fluid embolism, puerperal infection and hepatic diseases, and these 6 causes constituted 81.6% of the total. According to the audit of mortality cases demanded by WHO, the results showed that there was 73.0% avoidable deaths and 27.0% unavoidable deaths. The main problems were: 49.7% among the medical and health care system, chiefly lack of knowledge and skill of the medical personnels; 44.5% of the pregnant women and their family members lack the basic health knowledge. CONCLUSION: To improve the living condition of both mother and children, to increase hospital delivery rate, training medical personnel's skill, to provide better maternal health care, and to strengthen health education are effective measures to decrease MMR.

Cause of Death↗
PubMed1999

[Relationship of polymorphism of LMP2 gene to insulin-dependent diabetes mellitus and DR3 gene].

OBJECTIVE: To investigate the relationship of polymorphism of large multifunctional protease (LMP) 2 genes to insulin-dependent diabetes mellitus (IDDM) and DR3 gene. METHODS: The polymorphism of LMP2 genes was identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 68 IDDM patients and 71 healthy controls. IDDM patients and healthy controls were respectively divided into 2 groups according to DR3 genotyping. One group included subjects with DR3 gene, and another group subjects without DR3 gene. RESULTS: In the subjects with DR3 gene, the frequencies of LMP2-R/H, LMP2-R/R, LMP2-H/H were 47%, 47% and 6% in the IDDM patients, 13%, 87%, 0% in the controls. Compared with the Controls, the IDDM patients had a higher frequency of LMP2-R/H (P < 0.05) and a lower frequency of LMP2-R/R (P < 0.05). In the subjects without DR3 gene, the frequencies of LMP2-R/H, LMP2-R/R, LMP2-H/H were 58%, 39%, 2.8% respectively in the IDDM patients, and 32%, 54%, 14% respectively in the controls. Compared with the controls, the IDDM patients had a higher frequency of LMP2-R/H (P < 0.05), the frequency of LMP2-R/R in the IDDM patients did not have a significant difference from that in the controls (P > 0.05). CONCLUSIONS: There may be a significant association of LMP2-R/H and LMP2-R/R with the susceptibility to IDDM. The LMP2-R/H is a susceptible gene. The people with LMP2-R/H have a higher risk of suffering from IDDM. The LMP2-R/R is a protective gene. The people with LMP2-R/R have a decreased risk of suffering from IDDM. The effects of LMP2 on ID-DM are not affected by DR3 gene.

Cysteine Endopeptidases↗
PubMed1999

Quantitative assessment of Parkinson's disease deficits.

OBJECTIVE: To quantitatively analyze the tremor and rigidity due to Parkinson's disease. METHODS: 38 patients with Parkinson's disease (PD) ranging in age from 45 to 72 years and 211 normal subjects aged from 16 to 76 years were investigated. The frequency and range of tremor, the muscle tone of the upper limbs in elbow were detected by a computerized video motion detecting system and a new invented apparatus which can detects skeletal muscle tone. RESULTS: For the PD patients, the frequency of resting tremors was detected in 4 to 6 per second. For extensor and flexor in the PD patients, the value of muscle tone was higher than that of normal subjects and the value of muscle tone in flexor was higher than that of extensor. The rigidity increased gradually with repeat passive movement. The curves of rigidity were shown on computer screen or printed out. The data of rigidity were compared with the M-A Scale. A patient who was suspected to suffer from PD above by the equipments and found the muscle tone was higher than normal. In another PD patient the rigidity was obvious at one side and the muscle tone in "normal side" was also high. These equipments were used to record changes of rigidity and tremor in one more PD patient taking with different drugs in order to see the drug effect. CONCLUSION: Quantitative methods are useful to analyse the motion disorders due to PD.

Aged↗
PubMed1999

[Comparative study of conventional CT and spiral CT in diagnosis of lung cancer].

OBJECTIVE: To observe and compare the findings of lung cancer with conventional CT and spiral CT. METHODS: One hundred and thirty-four cases of pathologicalty confirmed lung cancer, including 56 centrally located and 78 peripherally located lung cancer, were examined by conventional CT and spiral CT, and their findings compared. RESULTS: In 30 of the 56 cases with central lung cancer, the lesion was located in the right bronchus and in 26 of them the lesion was in the left bronchus. On spiral CT, the display rate of obstructive findings and invasion to big vessels in the mediastinum was higher than that on conventional CT. There was no remarkable difference in density of the lesion and detection of metastasis in hilar and mediastinal lymph nodes between conventional and spiral CT. In 34 of the 78 cases with peripheral lung cancer, the lesion was in the right lung and in 44 was the lesion in the left lung. Calcification, air-bronchogram, cavity and pleural indentation of the lesion in the peripheral lung cancer were better displayed by spiral than by conventional CT. CONCLUSION: Spiral CT is superior to conventional CT in the observation of signs characteristic of central and peripheral lung cancers.

Adult↗
PubMed1999

[A study on homozygous deletion, hypermethylation, mutation and expression of p16 gene in human breast cancer].

OBJECTIVE: To investigate the homozygous deletion (HZD), hypermethylation and mutation of p16 gene in human breast cancer and the relationship between the structural alterations of p16 gene and its expression. METHODS: PCR and PCR-methylation assay with silver staining (PCR-MASS) were used to detect HZD and hypermethylation of p16 gene exon 1 in 60 fresh breast cancers and 24 normal breast tissues adjacent to cancer (as control tissues). PCR-SSCP and DNA sequencing were used for the analysis of p16 gene mutation. Moreover, p16 gene and mRNA were also detected in the 60 cases. RESULTS: Of the 60 breast cancers, HZD was found in 7 cases, hypermethylation and mutation were found in 16 and 4 cases respectively, and the difference was statistically significant. The positive rates of p16 protein and mRNA in breast cancer were 28/60 and 39/60 respectively. CONCLUSION: The results demonstrate that several kinds of p16 gene structural changes exist in breast cancer. The structural changes of p16 gene cause abnormal p16 expression, the main mechanisms are hypermethylation, while HZD or mutation are the secondary causes. Abnormal expression of p16 gene then becomes involved in the development and metastasis of breast cancer.

Breast Neoplasms↗
PubMed1998

Thiol-mediated disassembly and reassembly of [2Fe-2S] clusters in the redox-regulated transcription factor SoxR.

SoxR, a transcription factor containing [2Fe-2S] clusters, governs the cellular response to oxidative stress in Escherichia coli. The oxidation state of the iron-sulfur clusters regulates the SoxR transcriptional activity. When the reduced iron-sulfur clusters become oxidized ([2Fe-2S]2+ state), SoxR is activated to stimulate transcription of the soxS gene, whose product in turn switches on a group of genes encoding various proteins that defend against oxidative stress and antibiotics. A previous study showed that the oxidized [2Fe-2S] clusters of SoxR are destroyed by a free-radical-dependent process in vitro during aerobic exposure to the biological thiol glutathione. Here, we show that different thiols have differing effects on the SoxR [2Fe-2S] clusters. Like reduced glutathione, N-acetyl-L-cysteine, L-cysteine methyl ester, and L-cysteine ethyl ester disrupted the SoxR [2Fe-2S] clusters in aerobic solution. This disruption was blocked by L-cysteine, which was effective at concentrations 100-fold lower (1-10 microM) than the disrupting thiols (1 mM). In view of a previous observation that superoxide dismutase and catalase block the disruption process, this result suggests that L-cysteine may quench reactive SoxR or thiol intermediates involved in the cluster disruption reaction, the detailed mechanism of which remains unknown. In contrast, bifunctional thiols such as dithiothreitol or dithioerythritol promoted the aerobic assembly of the functional [2Fe-2S] clusters into apo-SoxR in the presence of Fe2+ and inorganic sulfide. The dithiol protein thioredoxin-A of E. coli acted catalytically in vitro in the presence of thioredoxin reductase and NADPH to promote [2Fe-2S] cluster assembly into apo-SoxR. The regulatory activity of SoxR in vivo, assessed by monitoring the paraquat-mediated induction of a soxS'::lacZ reporter fusion, was significantly lower in a strain lacking both thioredoxin-A and glutathione reductase, which maintains reduced glutaredoxins. Thus, cellular monothiols and dithiol proteins may contribute to SoxR regulation by affecting the disassembly and reassembly of the [2Fe-2S] clusters.

Apoproteins↗
PubMed1998

Important contributions of P-selectin glycoprotein ligand-1-mediated secondary capture to human monocyte adhesion to P-selectin, E-selectin, and TNF-alpha-activated endothelium under flow in vitro.

In this study, an in vitro flow model and a blocking mAb to P-selectin glycoprotein ligand-1 (PSGL-1) were used to define the role of PSGL-1 in monocyte attachment and rolling on E- and P-selectin and in attachment and accumulation on 6-h TNF-alpha-activated HUVEC. KPL1, an adhesion-blocking mAb directed against the tyrosine sulfate motif of PSGL-1, abolished monocyte-adhesive interactions with P-selectin, but only partially blocked monocyte interaction with E-selectin. Further analysis showed that on E-selectin, KPL1 blocked only secondary (i.e., monocyte/monocyte) interactions, but did not block primary (i.e., monocyte/E-selectin) interactions, with secondary adhesion accounting for 90% of the total adhesive interactions on either E- or P-selectin. On cytokine-activated HUVEC, monocytes initially attached and formed linear strings of adherent cells, which involved both primary and secondary adhesion. PSGL-1 or L-selectin mAb reduced string formation, and the combination of PSGL-1 and L-selectin mAb prevented monocyte strings and inhibited 86% of accumulation. Monocyte attachment and rolling on purified adherent monocytes were also critically dependent on PSGL-1 on the adherent monocytes. These studies document that secondary interactions between monocytes, mediated by PSGL-1, are crucial for monocyte initial attachment, rolling, and accumulation on activated endothelium under laminar shear flow.

Animals↗
PubMed1998

Smart X-ray beam position monitor system using artificial-intelligence methods for the Advanced Photon Source insertion-device beamlines.

At the Advanced Photon Source (APS), each insertion-device (ID) beamline front end has two X-ray beam position monitors (XBPMs) to monitor the X-ray beam position for both vertical and horizontal directions. Performance challenges for a conventional photoemission-type XBPM during operations are contamination of the signal from the neighbouring bending-magnet sources and the sensitivity of the XBPM to the insertion-device gap variations. Problems are exacerbated because users change the ID gap during their operations, and hence the percentage level of the contamination in the front-end XBPM signals varies. A smart XBPM system with a high-speed digital signal processor has been built at the Advanced Photon Source for the ID beamline front ends. The new version of the software, which uses an artificial-intelligence method, provides a self-learning and self-calibration capability to the smart XBPM system. The structure of and recent test results with the system are presented in this paper.

Journal Article↗
PubMed1998

A novel P-selectin glycoprotein ligand-1 monoclonal antibody recognizes an epitope within the tyrosine sulfate motif of human PSGL-1 and blocks recognition of both P- and L-selectin.

Interactions between P-selectin and P-selectin glycoprotein ligand-1 (PSGL-1) mediate the earliest "rolling" of leukocytes on the lumenal surface of endothelial cells at sites of inflammation. Previously, PSGL-1 has been shown to be the primary mediator of interactions between neutrophils and P-selectin, but studies on the ability of PSGL-1 to mediate interactions between P-selectin and other subsets of leukocytes have yielded variable and conflicting results. A novel IgG monoclonal antibody (MoAb) to human PSGL-1 was generated, and the specificity of this MoAb was confirmed by both flow cytometric analysis and Western blotting of cells transfected with human PSGL-1. This newly developed MoAb, KPL1, inhibited interactions between P-selectin expressing COS cells and either HL60 cells, neutrophils, or lymphocytes. Furthermore, KPL1 completely inhibited interactions between P-selectin and either purified CD4 T cells or neutrophils in a flow assay under physiological conditions, but had no effect on interactions of T cells or neutrophils with E-selectin. In addition, KPL1 blocked interactions between lymphoid cells transfected with L-selectin and COS cells expressing PSGL-1. The KPL1 epitope was mapped to a site within a consensus tyrosine sulfation motif of PSGL-1, previously shown to be essential for interaction with P-selectin and now shown to be essential for interaction with L-selectin, and to be distinct from the epitope identified by the PL1 function blocking anti-PSGL-1 MoAb. Two-color flow cytometry of normal leukocytes showed that while natural killer (NK) cells (CD16(+)), monocytes, CD4 and CD8 T cells, and alpha/beta and gamma/delta T cells were uniformly positive for PSGL-1, B cells expressed low levels of the KPL1 epitope. This low level of KPL1 staining was also observed immunohistologically in germinal centers, which had no detectable KPL1 staining, whereas T-cell areas (interfollicular region) were positive for KPL1. Interestingly, plasma cells in situ and interleukin-6-dependent myeloma cell lines were KPL1(+). Thus, PSGL-1 is expressed on essentially all blood neutrophils, NK cells, B cells, T cells, and monocytes. Variation in tyrosine sulfation during B-cell differentiation may affect the ability of B cells to interact with P- and L-selectin.

Animals↗
PubMed1998

Comparative neurotoxicity of oxaliplatin, cisplatin, and ormaplatin in a Wistar rat model.

Oxaliplatin (4 mg/kg), cisplatin (2 mg/kg with 20 mg/kg mannitol) and ormaplatin (2 mg/kg) were administered i.p. twice weekly for 4.5 weeks. Lactose injections (0.9%) were used as a control for oxaliplatin and 0.9% saline injections were used as a control for cisplatin and ormaplatin. Morphometric changes to dorsal root ganglia L4-L6 were quantitated as a measure of neurotoxicity. Drug treatment resulted in a decrease in cell and nuclear area and an increase in the percentage of cells with eccentric nucleoli for neuronal cell bodies in the DRG. Immediately following treatment the order of morphometric changes was ormaplatin > cisplatin > or = oxaliplatin. The accumulation of platinum in the DRG was measured by inductively coupled plasma mass spectrometry. The order of accumulation was cisplatin > oxaliplatin > ormaplatin. Following an 8-week recovery period the order of morphometric changes to the DRG was ormaplatin approximately equal to oxaliplatin > cisplatin. This correlated with a greater retention of platinum by the DRG for ormaplatin and oxaliplatin than for cisplatin. The results suggest that ormaplatin is uniquely neurotoxic immediately following treatment in the Wistar rat model. However, following an 8-week recovery period both ormaplatin and oxaliplatin are more neurotoxic than cisplatin and this neurotoxicity correlates with a greater retention of platinum by the DRG.

Animals↗
PubMed1998

IGF-I binding proteins, IGF-I binding protein mRNA and IGF-I receptor mRNA in rats with acute renal failure given IGF-I.

BACKGROUND: Recombinant human insulin-like growth factor-I (rhIGF-I) accelerates recovery from acute renal failure (ARF) in rats. IGF-I acts through the IGF-I receptor (IGF-IR) and its actions may be modified by IGF-I binding proteins (IGFBPs). It therefore would be of value to determine the effects of both ARF and rhIGF-I treatment on serum IGFBPs and mRNA for IGFBPs and IGF-IR. METHODS: Rats with ARF and sham-operated control rats were randomized to receive rhIGF-I or vehicle injections thrice daily for 72 to 74 hours starting five hours after surgery. Serum IGFPBs 1 to 6 were measured serially, and mRNA for IGFBPs 1 to 6 and for IGF-IR were measured in several tissues obtained 72 to 74 hours after surgery. RESULTS: At 72 to 74 hours, serum IGFBP-1 and IGFBP-2 levels were higher in rhIGF-I treated rats. Serum IGFBP-3 was affected by both ARF and rhIGF-I. IGFBP-4 rose transiently only in ARF groups. At 72 to 74 hours, mRNA for several IGFBPs was reduced in renal cortex of ARF rats. Low mRNA for IGFBP-4 and -6 was observed in renal medulla of the ARF rats, particularly in comparison to the sham-operated rats receiving vehicle. Renal medullary IGFBP-2 mRNA was decreased in ARF and sham rats given rhIGF-I as compared to sham animals given vehicle. Hepatic IGFBP-2 mRNA was higher in both rhIGF-I treated groups versus those given vehicle. Otherwise, there were no differences in IGFBP mRNAs among the four groups in lung, heart, and skeletal muscle. IGF-IR mRNA was decreased in renal cortex and medulla of both ARF groups and was not detected in liver in any group. CONCLUSIONS: Thus, ARF and rhIGF-I treatment each affected certain serum IGFBPs and jointly affected some IGFBPs. ARF suppressed gene transcription for renal cortical and medullary IGF-IR and some IGFBPs. rhIGF-I independently affected some renal cortical or medullary IGFBP mRNAs. rhIGF-I increased hepatic IGFBP-2 mRNA and serum IGFBP-2. These effects of ARF or rhIGF-I may influence rhIGF-I actions in rats with ischemic ARF.

Acute Kidney Injury↗
PubMed1998

Characterization of a double homeodomain protein (DUX1) encoded by a cDNA homologous to 3.3 kb dispersed repeated elements.

Target genes for the helicase-like transcription factor (HLTF), a member of the SNF/SWI family, were immunoprecipitated from HeLa chromatin fragments with an anti-HLTF antibody. A 182 bp fragment ( HEFT1 ) presented 87% sequence identity with 3.3 kb dispersed repeats from the 4q35 D4Z4 locus linked to facioscapulohumeral muscular dystrophy (FSHD). The HEFT1 loci were, however, not genetically linked to FSHD. Transfection and in vitro binding studies identified within HEFT1 a promoter whose basal activity required a GC box activated by Sp1 or Sp3. A 4.4 kb homologous transcript was found mostly in human skeletal muscle and heart. A 1.2 kb cDNA fragment was cloned that encoded a 170 amino acid protein (DUX1) with two paired-type homeodomains. In vitro translated DUX1 specifically interacted in electrophoretic mobility shift assay (EMSA) with a P5 oligonucleotide (5'-GATCTGAGTCTAATTGAGAATTACTGTAC-3'). DUX1 co-expression activated up to 5-fold transient expression in insect cells of a minimal promoter-luciferase construct fused to P5. The presence of 20 kDa DUX1 in vivo in rhabdomyosarcoma TE671 cell extracts was shown by western blotting with a rabbit antiserum raised against a DUX1 peptide. This antiserum suppressed a TE671 protein-P5 complex in EMSA with identical migration as the in vitro translated DUX1-P5 complex. Genomic PCR experiments could not identify a gene fragment linking the HEFT1 and DUX1 sequences, which present one mismatch in their overlapping region. However, a similar gene was found in another 3.3 kb element comprising the HEFT1 promoter and a DUX1 -like open reading frame. In addition, homologous gene sequences were identified in 3.3 kb elements of the D4Z4/FSHD locus, considered until now 'junk' DNA.

Amino Acid Sequence↗
PubMed1998

Measurement of Actinomyces pyogenes specific antibodies in bovine blood samples by an enzyme-linked immunosorbent assay.

In the present investigation Actinomyces pyogenes specific antigens caused an antibody response in the host. This could be determined by two enzyme-linked immunosorbent assays (ELISA) using cell extracts and a haemolysin preparation as antigens. An increased antibody titre was detectable in the sera of cows vaccinated with culture supernatants of A. pyogenes and Peptococcus indolicus, in the sera of cows infected with live cells of A. pyogenes and P. indolicus, in the sera of cows suffering from 'summer mastitis', and in part of the sera of nonvaccinated, apparently healthy cows. However, the titre of the cows that were vaccinated with culture supernatants decreased 8-9 months after inoculation. Because of the wide variation in antibody titre the determination of A. pyogenes specific antibodies seems to be only of limited use for the control of this infection.

Actinomyces↗