Methods for improved protein expression using pET vectors.
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Blotting, Western↗
Biomedical subjects
J Murray
Publications and source records attributed to J Murray.
Public health heresy.
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Public Health Administration↗
Three-dimensional motility cycle in leukocytes.
A 3-dimensional dynamic image analyzing system (3D-DIAS) has been developed in which a translocating cell is optically sectioned in the z-axis within a 2 sec period; the perimeter of the cell in each section is digitized into the 3D-DIAS data file, and the digitized perimeters are wrapped in order to reconstruct the cell image in three dimensions. Using 3D-DIAS, we have obtained the first dynamic 3-dimensional description of human polymorphonuclear leukocytes (PMN) translocating on a glass surface. A general behavior cycle has emerged which includes two phases. In the first, an ellipsoidal PMN with significant z-axis extends anteriorly and descends to the substratum. When the ventral surface of the anterior end contacts the substratum, there is rapid anterior expansion, which correlates with velocity peaks. In the second phase, the elongate PMN stops translocating along the substratum, the anterior end lifts off of the substratum, sometimes to heights greater than the length of the PMN at the substratum, and finally the PMN retracts into an ellipsoidal morphology still capable of random protrusions. During this second phase, which correlates with velocity troughs, turning usually occurs. The degree of turning is restricted by the continuous integrity of the posterior uropod. The period of the behavior cycle varies from roughly 0.5 to 2 min between PMNs, but is relatively constant within each individual PMN.
Cell Movement↗
Behavior of Dictyostelium amoebae is regulated primarily by the temporal dynamic of the natural cAMP wave.
The instantaneous velocity plots of Dictyostelium discoideum amoebae responding to natural waves and simulated temporal waves of cAMP with periods of 7 min are highly similar. This similarity has been used to deduce the dynamics of a natural wave crossing an amoeba, and the behavior of amoebae has been characterized during the different phases of a natural wave with a computer-assisted dynamic image analyzing system. During the first approximately 150 sec of the front of a natural wave, cells move persistently toward the aggregation center, with high instantaneous velocity and a decreased frequency of lateral pseudopod formation. During the last 30 sec of the front of the wave and the first 30 sec of the back of the wave, there is a "freeze" in cell shape and a dramatic depression in cell motility, pseudopod formation, and intracellular particle movement. During the last 180 sec of the back of the wave, there is a rebound in pseudopod formation, but it is random in direction and leads to no net cellular translocation. The data suggest that all of the behavior of a cell but orientation during the translocation phase is mediated by the temporal dynamics of the wave. The data also suggest that orientation toward the aggregation center occurs early in the front of the wave and that, once oriented, cells move in a blind fashion during the translocation phase.
Animals↗
NO and NO accommodation.
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Adaptation, Physiological↗
Patient education: recommendations regarding sunscreens, drugs, and diet.
Early recognition of melanoma is directly related to improvement in survival. Patients, therefore, must not only be educated in recognition of abnormal skin lesions, but also in proper skin examination, ultraviolet radiation protection, effect of drugs on the development of malignancies, and dietary means of promoting wellness and preventing disease. Basic patient instruction should begin at the time of diagnosis and should include all health care team members to assure patient comprehension and compliance with recommendations.
Dietary Fats↗
Beta-adrenergic neuroeffector abnormalities in the failing human heart are produced by local rather than systemic mechanisms.
In order to investigate the general cause of beta-adrenergic receptor neuroeffector abnormalities in the failing human heart, we measured ventricular myocardial adrenergic receptors, adrenergic neurotransmitters, and beta-adrenergic receptor-effector responses in nonfailing and failing hearts taken from nonfailing organ donors, subjects with endstage biventricular failure due to idiopathic dilated cardiomyopathy (IDC), and subjects with primary pulmonary hypertension (PPH) who exhibited isolated right ventricular failure. Relative to nonfailing PPH left ventricles, failing PPH right ventricles exhibited (a) markedly decreased beta 1-adrenergic receptor density, (b) marked depletion of tissue norepinephrine and neuropeptide Y, (c) decreased adenylate cyclase stimulation in response to the beta agonists isoproterenol and zinterol, and (d) decreased adenylate cyclase stimulation in response to Gpp(NH)p and forskolin. These abnormalities were directionally similar to, but generally more pronounced than, corresponding findings in failing IDC right ventricles, whereas values for these parameters in nonfailing left ventricles of PPH subjects were similar to values in the nonfailing left ventricles of organ donors. Additionally, relative to paired nonfailing PPH left ventricles and nonfailing right ventricles from organ donors, failing right ventricles from PPH subjects exhibited decreased adenylate cyclase stimulation by MnCl2. These data indicate that: (a) Adrenergic neuroeffector abnormalities present in the failing human heart are due to local mechanisms; systemic processes do not produce beta-adrenergic neuroeffector abnormalities. (b) Pressure-overloaded failing right ventricles of PPH subjects exhibit decreased activity of the catalytic subunit of adenylate cyclase, an abnormality not previously described in the failing human heart.
Adenylyl Cyclases↗
Concordance of the MCMI-II, the MMPI, and Axis I discharge diagnosis in psychiatric inpatients.
Discriminant functions of the MMPI and the MCMI-II were compared in a sample of 166 hospitalized psychiatric patients with discharge diagnoses of affective disorder (63), schizophrenia (26), substance abuse (35), and other disorders (42). Of special interest was the comparative diagnostic utility of the two instruments in regards to DSM-III-R Axis I diagnoses. Both tests performed reasonably well in the discriminant function analyses; however, the MCMI-II achieved a somewhat superior overall hit rate with this sample of inpatients (79% to 68%). This difference was tied to greater accuracy of the MCMI-II for identifying the affective disorders group.
Adolescent↗
Cocaine/polydrug use in pregnancy: two-year follow-up.
The impact of cocaine on pregnancy and neonatal outcome has been well documented over the past few years, but little information regarding long-term outcome of the passively exposed infants has been available. In the present study, the 2-year growth and developmental outcome for three groups of infants is presented: group 1 infants exposed to cocaine and usually marijuana and/or alcohol (n = 106), group 2 infants exposed to marijuana and/or alcohol but no cocaine (n = 45), and group 3 infants exposed to no drugs during pregnancy. All three groups were similar in racial and demographic characteristics and received prenatal care through a comprehensive drug treatment and follow-up program for addicted pregnant women and their infants. The group 1 infants demonstrated significant decreases in birth weight, length, and head circumference, but by a year of age had caught up in mean length and weight compared with control infants. The group 2 infants exhibited only decreased head circumference at birth. Head size in the two drug-exposed groups remained significantly smaller than in control infants through 2 years of age. On the Bayley Scales of Infant Development, mean developmental scores of the two groups of drug-exposed infants did not vary significantly from the control group, although an increased proportion of group 1 and 2 infants scored greater than two standard deviations below the standardized mean score on both the Mental Developmental Index and the Psychomotor Developmental Index compared with the control infants. Cocaine exposure was found to be the single best predictor of head circumference.(ABSTRACT TRUNCATED AT 250 WORDS)
Child, Preschool↗
Regional mapping of facioscapulohumeral muscular dystrophy gene on 4q35: combined analysis of an international consortium.
Members of an international consortium for linkage analysis of the facioscapulohumeral muscular dystrophy (FSHD) gene have pooled data for joint analyses, in an attempt to determine the precise location of the FSHD gene and the order of four DNA markers on 4q35 region. Six laboratories determined a total of 3,078 genotypes in 65 families, consisting of a total of 504 affected subjects and 559 unaffected subjects. For each marker, a mean of 648 meioses were informative. D4S139 and D4S163 were identified as the closest linked markers to the FSHD locus, with 99% upper confidence intervals of recombination fractions of .08 and .10, respectively. We have used the CRI-MAP program to construct the most likely order of cen-D4S171-F11-D4S163-D4S139-FSHD-tel, with favorable odds of 10(8)-10(114) over all other orders except that in which F11 and D4S171 are reversed, for which the odds ratio was 191:1. With this order, the genetic map of this region extends 25.5 cM in males and 13.8 cM in females (averaging 19.5 cM for sexes combined); the sex difference was statistically significant (P = .0013). Comparison between families for the two-point and multipoint lod scores involving FSHD showed no evidence for heterogeneity of this disorder. However, after the completion of this analysis, one large family which might show heterogeneity was identified. In view of this and the fact that all of the linked markers reside on the same side of the FSHD locus, the clinical application of these markers is not recommended at this time.
Chromosome Mapping↗
Dentin phosphoprotein gene locus is not associated with dentinogenesis imperfecta types II and III.
Dentinogenesis imperfecta (DGI) is an autosomal dominant inherited dental disease which affects dentin production and mineralization. Genetic linkage studies have been performed on several multigeneration informative kindreds. These studies determined linkage between DGI type II and III and group-specific component (vitamin D-binding protein). This gene locus has been localized to the long arm of human chromosome 4 in the region 4q11-q21. Although this disease has been mapped to chromosome 4, the defective gene product is yet to be determined. Biochemical studies have suggested abnormal levels of dentin phosphoprotein (DPP) associated with DGI type II. This highly acidic protein is the major noncollagenous component of dentin, being solely expressed by the ectomesenchymal derived odontoblast cells of the tooth. The purpose of the present study was to establish whether DPP is associated with DGI types II and III, by using molecular biology techniques. The strategy was to use a synthetic degenerative DPP oligonucleotide probe to map this sequence to the long arm of human chromosome 4, 4q13-q21, by using somatic cell hybrids. Our results indicated that DPP is not localized to any region of human chromosome 4, thus suggesting that the DPP gene is not directly associated with DGI type II or DGI type III. Our data do not exclude the possibility that other proteins associated with DPP posttranslational modifications might be responsible for this genetic disease.
Adult↗
Cloning and characterization of the rad4 gene of Schizosaccharomyces pombe; a gene showing short regions of sequence similarity to the human XRCC1 gene.
The rad4.116 mutant of the fission yeast Schizosaccharomyces pombe is temperature-sensitive for growth, as well as being sensitive to the killing actions of both ultraviolet light and ionizing radiation. We have cloned the rad4 gene by complementation of the temperature sensitive phenotype of the rad4.116 mutant with a S. pombe gene bank. The rad4 gene fully complemented the UV sensitivity of the rad4.116 mutant. The gene is predicted to encode a protein of 579 amino acids with a basic tail, a possible zinc finger and a nuclear location signal. The amino terminal part of the predicted rad4 ORF contains two short regions of similarity to the C-terminal part of the human XRCC1 gene. Codon usage suggests that the gene is very poorly expressed, and this was confirmed by RNA studies. Gene disruption showed that the rad4 gene was essential for the mitotic growth of S. pombe.
Base Sequence↗
The prevalence and age distribution of peripheral pulmonary hamartomas in adult males. An autopsy-based study.
This autopsy-based study defined the prevalence and age distribution of peripheral pulmonary hamartomas in 47,635 southern African miners examined between 1975 and 1988. The prevalence rate for white miners was 7.5/1,000 and for black miners 1.1/1,000. When directly standardised to the white men in the general population, the rates for white and black miners were 7.2 and 5.5/1,000, respectively. The prevalence of peripheral pulmonary hamartomas in both groups increased with age, from 0.8/1,000 in the third decade to 12.0/1,000 in the eighth decade. The study showed a much higher prevalence of peripheral pulmonary hamartomas for whites than previously reported. Furthermore, it documented the occurrence of these benign lung tumours in blacks, a fact that has previously been questioned. There appeared to be no significant difference in prevalence and age distribution between white and black miners, although the database for the black group was deficient for the later decades of life.
Adult↗
Myosin IB null mutants of Dictyostelium exhibit abnormalities in motility.
Cellular and intracellular motility are compared between normal Dictyostelium amoebae and amoebae lacking myosin IB (DMIB-). DMIB- cells generate elongated cell shapes, form particulate-free pseudopodia filled with F-actin, and exhibit an anterior bias in pseudopod extension in a fashion similar to normal amoebae. DMIB- cells also exhibit a normal response to the addition of the chemoattractant cAMP, including a depression in cellular and intracellular particle velocity, depolymerization of F-actin in pseudopodia, and a concomitant increase in cortical F-actin. DMIB- cells do, however, form lateral pseudopodia roughly three times as frequently as normal cells, turn more often, and exhibit depressed average instantaneous cell velocity. DMIB- cells also exhibit a decrease in the average instantaneous velocity of intracellular particle movement and an increase in the degree of randomness in particle direction. These findings indicate that if there is functional substitution for myosin IB by other myosin I isoforms, it is at best only partial, with myosin IB being necessary for maintenance of the normal rate and persistence of cellular translocation, suppression of lateral pseudopod formation and subsequent turning, rapid intracellular particle motility, and the normal anterograde bias of intracellular particle movement. Furthermore, it is likely that the behavioral abnormalities observed here for DMIB- cells underlie the delay in the onset of chemotactic aggregation, the increase in the time required to complete streaming, and the abnormalities in morphogenesis exhibited by DMIB- cells.
Actins↗
Assignment of ten DNA repair genes from Schizosaccharomyces pombe to chromosomal NotI restriction fragments.
Ten DNA repair (rad) genes from the fission yeast, Schizosaccharomyces pombe were mapped to the 17 NotI fragments of the three chromosomes. Nine of the genes map to chromosome I, but there is no evidence for significant clustering.
Chromosome Mapping↗
Granulomatous gastritis: part of a vasculitic syndrome.
The previously unreported association of granulomatous gastritis and mononeuritis multiplex occurring in the setting of a vasculitic syndrome is described. The two conditions are considered to be associated and to be immune mediated. The previously accepted concept of isolated granulomatous gastritis is disputed.
Female↗
Percutaneous urinary diversion in gynecologic oncology.
During the 10-year period after 1979, percutaneous urinary diversion (PCUD) was performed on 35 patients whose mean age was 53.5 years (30-80 years). Twenty-one patients (60%) had Stage IIB-IV cervical cancer, 11 (31%) Stage IB-IIA cervical cancer, 2 (6%) Stage IB-II endometrial cancer, and 1 (3%) Stage IB vaginal cancer. All had radiological evidence of ureteric obstruction and 8 patients also had urinary tract fistulae. Serum creatinine levels were elevated in 24. Following diversion there was a significant fall in mean pretreatment creatinine levels from 482 mumol/liter (range, 70-1703 mumol/liter) to 131 mumol/liter (range, 60-290 mumol/liter; P less than 0.0001); those patients with normal creatinine levels prior to diversion also had a reduction in their levels. A significant fall in mean serum urea levels from 22.0 mmol/liter pre- to 11.9 mmol/liter post-PCUD (P less than 0.001) was also noted. Minor complications occurred and included hemorrhage, replacement/reinsertion, infection, and blockage. Median survival of the 35 patients after PCUD was 6 months (mean, 16.5 months). For the 11 with normal pretreatment renal function median survival was 16 months (mean, 41 months) compared to 2.5 months (mean, 5.1 months) for those with elevated pretreatment serum creatinine levels. Median survival with untreated malignancy was 7 months (mean, 19.6 months) and 6 months (mean, 12.3 months) in patients with previously treated cancer. PCUD is indicated in previously untreated patients with gynecologic cancer so that primary therapy can be instituted. The role of urinary diversion in patients with previously treated cancer must be individualized. Palliative diversion is appropriate in selected patients where additional therapy is expected to prolong life, where symptom control is needed, or to allow the patient to return home for a significant proportion of the remainder of life.
Adult↗
Rosai-Dorfman disease of the uterine cervix.
A case of Rosai-Dorfman disease of the uterine cervix is described. The disease is characterized by a mixed chronic inflammatory cell infiltrate in which histiocytic cells predominate. A characteristic feature is the presence within many of the histiocytic cells of engulfed, but undamaged, lymphocytes. Rosai-Dorfman disease is of unknown etiology and pursues a benign course. The disease can occur at a wide variety of extranodal sites, but this appears to be the first reported instance of female genital tract involvement.
Adult↗