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Biomedical subjects

J He

Publications and source records attributed to J He.

At least 379 records · Page 21Linked to original sources

In vitro transcription and binding analysis of promoter regulation by a host-specific signal in a phytopathogenic fungus.

The PDA1 promoter of the phytopathogenNectria haematococca MPVI (anamorph Fusarium solani) offers a model for regulation of a fungal virulence gene in response to plant host-specific signals. Expression of the PDA1 gene, encoding pisatin demethylase, is induced in culture by pisatin, the isoflavanoid phytoalexin of pea. This pisatin induction is suppressed by nutritional factors. We have been studying the mechanism of pisatin induction through in vitro identification of regulatory factors and regulatory elements of the PDA1 promoter. We have developed an in vitro transcription system for N. haematococca which accurately initiates at the PDA1 promoter and reflects the pisatin induction of PDA1 mRNA observed in vivo. This in vitro activity allowed a functional test of a limited set of 5' upstream deletions in the PDA1 promoter. In vitro binding studies have identified a DNA binding factor which is appears in mycelial extract after treatment of the mycelium with pisatin. This pisatin-responsive factor binds to a minimum size region of 35 bp approximately 500 bp upstream of the transcription initiation site. Tests using the in vitro transcription assay and in vivo competition both indicate a role for this binding region in the high expression of PDA1 under pisatin-induced conditions. Southwestern blotting has identified one component of this binding activity to be a approximately 35 kDa protein. The availability of these functional and structural tests of function, in conjunction with complementary in vivo tests, allow the detailed dissection of the signal pathway leading from exposure of the cell to pisatin towards the activation of PDA1 transcription.

Animals↗

Molecular basis of antigenic variation in infectious bursal disease virus.

Four antigenically different strains of infectious bursal disease virus (IBDV), characterized by their reactivities with a panel of neutralizing monoclonal antibodies (MAbs), were selected to determine the molecular basis of antigenic variation. The large genome segment A, encoding the structural proteins of the U.S. variants GLS, DS326, E/Del and the vaccine strain D78, was cloned and sequenced. Comparison of the deduced amino acid sequences of the U.S. variants with other IBDV strains showed that most of the amino acid substitutions occur in the central region between residues 212 to 332, especially in the two hydrophilic regions between residues 212 to 223 and residues 314 to 324 of VP2 protein. By comparing the amino acid sequences of these variant viruses and their reactivities with IBDV specific MAbs, the putative amino acids involved in the formation of virus-neutralizing epitopes were identified. Comparison of the D78 versus PBG98 sequence showed that Gln at position 249 (Gln249) appears to be critical in binding with MAb B69. Similarly, comparison of the U.S. variant sequences with other serotype 1 sequences showed unique substitution(s) at residue Glu321 in GLS, residues Ile286, Asp318, Glu323 in E/Del, and residues Glu311 and Gln320 in DS326, which could be potential residue(s) involved in the recognition of MAb57, MAb67, and MAb179 epitopes, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Expression of hepatitis E virus putative structural proteins in recombinant vaccinia viruses.

Hepatitis E virus (HEV) is a polyadenylated, positive-stranded RNA virus which is a major cause of enterically transmitted non-A, non-B hepatitis in many developing countries. The viral genome contains three different open reading frames (ORFs): ORF1, which is believed to encode nonstructural proteins, and ORF2 and ORF3, which are believed to encode structural proteins. The full-length putative structural proteins encoded by ORF2 and ORF3 of HEV have been cloned and expressed in recombinant vaccinia virus. Proteins encoded by ORF2 and ORF3 when expressed in vaccinia virus are recognized by pooled sera obtained from individuals with acute hepatitis E. Vaccinia-expressed viral gene products of HEV will have utility in characterizing the cell-mediated immune response to HEV.

Base Sequence↗

Chemical and mutagenic patterns of airborne particulate matter collected in 17 Italian towns.

The mutagenicity of airborne particulate matter collected in 17 towns of Italy in 1990 was assessed using the Ames test. The mutagenicity of crude extract correlated with amount of lead, suggesting the direct contribution of gasoline car exhausts. Moreover, the mutagenicity correlated with particulate matter amounts. An inverse correlation with temperature was observed. The crude extracts were fractionated in acid, basic, and neutral fractions. The latter was further separated into polycyclic aromatic hydrocarbon (PAH), polar, and nonpolar fractions. Acid and polar fractions showed the higher mutagenicity. Average recovery of mutagenicity was about 60%.

Air Pollutants↗

[Studies of enzyme-linked immunoassay for testosterone using penicillinase as label].

An enzyme-linked immunoassay (EIA) for testosterone was developed based on penicillinase as the marker enzyme. The assay proposed requires a conjugate containing penicillinase as the enzymatic component, penicillin Vas substrate and an iodine-starch reagent as the chromogenic component. The binding of conjugate molecules results in the transition of the iodic complex from dark blue to colourless. delta A (the difference between the blank and the testosterone) vs log C was linear over the range 10(-7) - 10(-2)mg/ml (r = 0.9969), with a limit of detection 25 pg per disc. The CVs of day to day and within day for plasma testosterone were < 3.3% and 5.9%, respectively. There was a good correlation between the values obtained by EIA and radioimmunoassay (r = 0.937).

Enzyme-Linked Immunosorbent Assay↗

Blood flow velocity in common carotid artery in humans during breath-holding and face immersion.

Blood flow velocity was measured in the common carotid artery of humans during breath-holding (BH) in the supine and upright positions, and during breath-hold face immersion (FIBH) in cold (20 degrees C) and warm (35 degrees C) water in the upright position with 90 degrees flexion of the upper body, to test the influences of the body position, facial immersion and temperature on changes in cerebral blood flow. Simultaneously, the heart rate (HR) and blood pressure (BP) under the same conditions, and the end-tidal PO2 and PCO2 (PETO2 and PETCO2) during supine BH, were also measured. Results showed that posture had no significant effect on the apneic HR. Bradycardia developed during cold FIBH, but not warm FIBH. The BP increased during BH in both positions, and during both cold and warm FIBH. The PETO2 decreased and the PETCO2 increased significantly at the end of BH. The blood velocity decreased transiently at the beginning of upright BH, but increased time-dependently during both supine and upright BH. Moreover, the velocity increased more during warm FIBH than during BH's, and even more during cold than warm FIBH. When bradycardia occurred during cold FIBH, the velocity integral per minute tended to decrease. These results suggest that the cerebral blood flow increases in these conditions of BH. This increase can be explained by the increase in PETCO2 and decrease in PETO2, possibly with a slight contribution from the increase in the BP.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Detection of specific DNA fragment of Plasmodium falciparum by solid phase PCR].

Solid phase PCR for detecting Plasmodium falciparum was established and used to detect cultured FCCl/HN isolate and malaria patients in Yunnan Province. The results revealed that the sensitivity of the method for detecting FCCl/HN isolate DNA was as minimal as 0.2 pg, or about 10 parasites. The specificity of the method was confirmed by discriminating malaria patients infected with different species of Plasmodium. The results suggest that solid phase PCR is a specific and sensitive method for early detection of P. falciparum.

Animals↗

Flow velocity in carotid artery in humans during immersions and underwater swimming.

Heart rate and flow velocity in the common carotid artery were examined on eight subjects in various underwater conditions. The heart rate decreased more markedly during whole-body immersion than during head-out immersion. The end-diastolic flow velocity and the flow-velocity integral in a cardiac cycle during whole-body immersion increased more markedly than during head-out immersion. Differences in heart rate and flow velocity were not detected between sitting and prone positions in whole-body immersion. Heart rate increased at the beginning and after underwater swimming. The peak systolic flow velocity increased significantly after the end of underwater swimming. The flow-velocity integral in a cardiac cycle decreased during underwater swimming, but in a minute did not change significantly. These results suggest that facial immersion is important in eliciting pronounced carotid artery flow-velocity response in addition to the heart-rate response and that underwater exercise influences these responses. However, underwater posture apparently does not influence them.

Adult↗

Localization of chromosome 9p homozygous deletions in glioma cell lines with markers constituting a continuous linkage group.

Southern blot analyses of the 9p-localized type I interferon (IFN) genes in DNAs obtained from malignant glioma cell lines and glial tumor tissue have indicated that homozygous deletions of the IFN-alpha and IFN-beta genes often occur during the development of the highly malignant central nervous system neoplasm, glioblastoma. We have applied a set of markers that span the IFN region on 9p to the analysis of DNAs from 30 human glioma cell lines in order to define the region of homozygous deletion associated with this cancer more precisely. Fourteen of the cell lines revealed either complete (12 cases) or partial (2 cases) homozygous deletions of the IFN-alpha gene cluster; no instances of homozygous deletions were observed that did not involve the IFN-alpha region. Genomic DNA identified by the markers nearest to and flanking the IFN-alpha genes were retained in 5 of the cases with homozygous deletions. Consequently, these results limit the extent of homozygous deletions in glioma cell lines to a small region of 9p21-p22 that includes most of the type I IFN locus.

Base Sequence↗

Autoimmune disease in mice due to integration of an endogenous retrovirus in an apoptosis gene.

The MRL-lpr/lpr mouse strain is a model of systemic autoimmune disease. In this model, intrinsic defects of intrathymic T cell development include defective deletion of self-reactive T cells and expression of endogenous retroviruses. Defective deletion of self-reactive T cells in the thymus has been proposed to be due to germline mutation in the Fas apoptosis gene. Using different fragments of a Fas cDNA probe, we determined that the lpr/lpr mutation is a 5.3-kb insertion of DNA within the second intron of the Fas gene. cDNA corresponding to this region was then derived from thymic RNA from MRL-lpr/lpr and MRL- +/+ mice using the polymerase chain reaction. All thymic RNA samples from MRL-lpr/lpr mice yielded a unique product that was 168 bp larger than that of MRL- +/+ mice. Complete sequence analysis indicated that this inserted sequence had 98% homology with a sequence from the 3' long terminal repeat of the early transposon (ETn). RNA analysis indicated higher expression of ETn RNA in the thymus of MRL-lpr/lpr than MRL- +/+ mice. The interdependence of ETn expression and abnormal Fas expression was then analyzed in a CD2-Fas transgenic mouse model in which a full-length murine Fas cDNA under the regulation of the CD2 promoter and enhancer was used to correct defective Fas expression in T cells of MRL-lpr/lpr mice. In these mice, reduced thymic ETn expression was observed, confirming that high ETn expression is related to abnormal Fas expression. These results establish a link between endogenous retrovirus expression, abnormal Fas expression, and autoimmune disease.

Amino Acid Sequence↗

Agreement between overnight and 24-hour urinary cation excretions in southern Chinese men.

Agreement between overnight and 24-hour urinary sodium, potassium, calcium, and magnesium excretion was studied in a sample of 63 normotensive Southwestern Chinese men: 30 Yi farmers and 33 urban residents in April 1989. Overnight (8-hour) and 24-hour urine specimens were collected on 3 consecutive days. Estimated correlation coefficients between 24-hour and overnight mean true values were 0.863 and 0.906 for sodium, 0.736 and 0.816 for potassium, 0.902 and 0.725 for calcium, and 0.733 and 0.703 for magnesium in Yi farmers and urban residents, respectively. Hourly overnight urinary sodium and potassium excretion rates were significantly lower than the corresponding hourly 24-hour urinary excretion rates: -0.60 and -1.99 mmol/hour for sodium, -1.24 and -0.48 mmol/hour for potassium (all p < 0.05) in Yi farmers and urban residents, respectively. In multiple regression analyses, the differences between 24-hour and overnight urinary sodium and potassium excretion rates were significantly and positively related to differences between 24-hour and overnight creatinine excretion rates. The ratios of intraindividual to interindividual variance were lower for 24-hour collections than for overnight collections for sodium and calcium, but the differences in these ratios for potassium and magnesium were small. For sodium and calcium, twice as many overnight as 24-hour collections were required to estimate the correlation between cations and blood pressure with the same accuracy; for potassium and magnesium, overnight and 24-hour collections were equally accurate. These results indicate that in normotensive populations such as the one studied, overnight urine collections may be used to estimate 24-hour cation excretion. The underestimate of cation excretion by assessments based on collection of overnight specimens may be due to either a lower creatinine clearance or a lower intake of cations at night.

Adult↗

Serum cholesterol in young men and subsequent cardiovascular disease.

BACKGROUND: The increased risk of cardiovascular disease associated with higher serum cholesterol levels in middle-aged persons has been clearly established, but there have been few opportunities to examine a potential link between serum cholesterol levels measured in young men and clinically evident premature cardiovascular disease later in life. METHODS: We performed a prospective study of 1017 young men (mean age, 22 years) followed for 27 to 42 years to quantify the risk of cardiovascular disease and total mortality associated with serum cholesterol levels during early adult life. The mean serum cholesterol level at entry was 192 mg per deciliter (5.0 mmol per liter). RESULTS: During a median follow-up of 30.5 years, there were 125 cardiovascular-disease events, 97 of which were due to coronary heart disease. The serum cholesterol level at base line was strongly associated with the incidence of events related to coronary heart disease and cardiovascular disease, as well as to total mortality and mortality due to cardiovascular disease. The risks were similar whether the events occurred before or after the age of 50. In a proportional-hazards analysis adjusted for age, body-mass index (the weight in kilograms divided by the square of the height in meters), the level of physical activity, coffee intake, change in smoking status, and the incidence of diabetes and hypertension during follow-up, a difference in the serum cholesterol level at base line of 36 mg per deciliter (0.9 mmol per liter)--the difference between the 25th and 75th percentiles of cholesterol level in the study population at base line--was associated with an increased risk of cardiovascular disease (relative risk, 1.72; 95 percent confidence interval, 1.39 to 2.14), coronary heart disease (relative risk, 2.01; 95 percent confidence interval, 1.59 to 2.53), and mortality due to cardiovascular disease (relative risk, 2.02; 95 percent confidence interval, 1.23 to 3.32). A difference in the base-line serum cholesterol level of 36 mg per deciliter was significantly associated with an increased risk of death before the age of 50 (relative risk, 1.64; 95 percent confidence interval, 1.03 to 2.61), but not with the overall risk of death (relative risk, 1.21; 95 percent confidence interval, 0.93 to 1.58). CONCLUSIONS: These findings indicate a strong association between the serum cholesterol level measured early in adult life in men and cardiovascular disease in midlife.

Adult↗

Direct and indirect effects of valinomycin upon cytochrome c oxidase.

Valinomycin binds beef heart cytochrome c oxidase with approximately equimolar stoichiometry (one valinomycin per cytochrome aa3 complex) and a Kd of 2.0-3.0 microM, almost independent of ionic strength, but dependent upon the presence of K+ ions. Its effects are twofold: a red shift in the Soret maximum of the resting enzyme from 419 to 422 nm, with an appreciable delta EmM at 430-410 nm, and an inhibition of catalytic activity of about 50%. Much smaller concentrations of valinomycin are sufficient to induce the release of respiration in cytochrome oxidase vesicles by nigericin. Intermediate levels of valinomycin alone are capable of partially releasing respiratory control, probably by increasing H+ permeability as well as that of K+. The amounts of valinomycin required to bind the enzyme are much greater than those required to collapse delta psi in vesicular systems in a steady state or during proton pulse experiments. We conclude that the two effects--a direct one upon the enzyme and an indirect one via abolition of electrogenic potentials--are of separate origins. The existence of the direct effect does not call for reinterpretation of classical results showing proton translocation, which are adequately explained by the indirect action of valinomycin as an ionophore.

Animals↗

Changes in carotid blood flow and electrocardiogram in humans during and after walking on a treadmill.

Blood flow velocity in the common carotid artery and the electrocardiogram were measured simultaneously by telemetry in seven male subjects during 20-min walking on a treadmill at an exercise intensity corresponding to a mean oxygen uptake of 26.0 (SD 2.9) ml.kg-1.min-1. The mean cardiac cycle was shortened from 0.814 (SD 0.103) s to 0.452 (SD 0.054) s during this exercise. Of this shortening, 73% was due to shortening of the diastolic period and 27% to shortening of the systolic period. In the relatively small shortening of the mean systolic period [from 0.377 (SD 0.043) s to 0.268 (SD 0.029) s], the isovolumetric contraction time was shortened by 56%. During exercise, the heart rate (fc) increased by 79.4% [from 74.3 (SD 9.3) beats.min-1 to 133.3 (SD 14.8) beats.min-1], and the peak blood velocity (S1) in the common carotid artery increased by 56.1% [from 0.82 (SD 0.10) m.s-1 to 1.28 (SD 0.11) m.s-1]. After exercise, the S1 decreased rapidly to the resting level. The fc decreased more slowly, still being higher than the initial resting level 5 min after exercise. The diastolic velocity wave and the end-diastolic foot decreased during exercise. The blood flow rate in the carotid artery increased transiently by 13.5% at the beginning of exercise [from 5.62 (SD 0.63) ml.s-1 to 6.38 (SD 0.85) ml.s-1] and by 26.5% at the end of the exercise period [from 5.62 (SD 0.63) ml.s-1 to 7.11 (SD 1.34) ml.s-1].(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Changes in cardiac rhythm in man during underwater submersion and swimming studied by ECG telemetry.

A previously reported method for electrocardiographic (ECG) telemetry in water using frequency-modulated current was improved to obtain more stable ECGs. The ECGs of seven healthy men were monitored using the improved method during and after whole-body submersion or underwater swimming. Bradycardia and arrhythmias were observed during the submersion, and transient tachycardia was detected after the start of underwater swimming, followed by bradycardia with arrhythmias. Three different types of arrhythmias were observed: sinus arrhythmia (SA), supraventricular extrasystole (SE) and ventricular extrasystole (VE). SA and SE tended to develop during the latter half of the period of submersion or underwater swimming, and especially after the restart of breathing. VEs were detected in only one subject during submersion, whereas they occurred in most subjects during and after underwater swimming. Individual variations were found in development of arrhythmias, one subject showing no arrhythmia. Bradycardia, SA and SE could depend on vagal suppression in underwater conditions, and VE may be related to the effect of muscular movement on cardiac function in addition to vagal inhibition.

Adult↗

Liquid holding effects on methylmercury genotoxicity in human lymphocytes.

Methylmercury chloride (MMC) treatment of resting (G0) human lymphocytes resulted in the formation of chromosome and chromatid aberrations. This treatment also induced aneuploidy in second metaphases, suggesting that MMC produces stable damage involved in chromosome segregation errors. The storage of treated cells (liquid holding for 48 h before cell proliferation) did not result in an important recovery from induced cell toxicity or chromosome damage. Therefore, MMC seems to be an X-ray-like agent, able to produce long-lasting damages giving rise to both structural and numerical chromosome abnormalities.

Aneuploidy↗

Validity of physicians' self-reports of cardiovascular disease risk factors.

To determine the accuracy of self-reported risk factors in 78 physicians, self-reported information was compared to findings on a standardized examination. Measured weight (r = 0.98), height (r = 0.95), body mass index (r = 0.96), systolic blood pressure (SBP) (r = 0.72), and diastolic blood pressure (DBP) (r = 0.60) were highly correlated with self-reported values (all P < 0.0001). Mean self-reported SBP and DBP did not differ from measured values; measured weight was 1.5 kg greater and measured height 1.4 cm less than self-reported values (both p < 0.0001). Regression of measured on self-reported values indicated excellent agreement except for DBP and heart rate. Differences between measured and self-reported values were not associated with a variety of variables except for a greater difference in SBP at higher levels of SBP. None of the 60 self-reported nonsmokers had expired carbon monoxide levels greater than 10 ppm. These results indicate that physicians' self-reports of height, body mass index, SBP, and smoking are extremely accurate and suitable for research purposes.

Aged↗