Perosomus elumbis in a Holstein calf in Brazil.
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Abnormalities, Multiple↗
Biomedical subjects
M A Romano
Publications and source records attributed to M A Romano.
Activation of MHC class I, II, and CD40 gene expression by histone deacetylase inhibitors.
Epigenetic mechanisms are involved in regulating chromatin structure and gene expression through repression. In this study, we show that histone deacetylase inhibitors (DAIs) that alter the acetylation of histones in chromatin enhance the expression of several genes on tumor cells including: MHC class I, II, and the costimulatory molecule CD40. Enhanced transcription results in a significant increase in protein expression on the tumor cell surface, and expression can be elicited on some tumors that are unresponsive to IFN-gamma. The magnitude of induction of these genes cannot be explained by the effect of DAIs on the cell cycle or enhanced apoptosis. Induction of class II genes by DAIs was accompanied by activation of a repressed class II transactivator gene in a plasma cell tumor but, in several other tumor cell lines, class II was induced in the apparent absence of class II transactivator transcripts. These findings also suggest that the abnormalities observed in some tumors in the expression of genes critical to tumor immunity may result from epigenetic alterations in chromatin and gene regulation in addition to well-established mutational mechanisms.
Animals↗
Evidence for and consequences of chronic heme deficiency in Belgrade rat reticulocytes.
The Belgrade rat has a microcytic, hypochromic anemia inherited as an autosomal recessive trait (gene symbol b). Transferrin-dependent iron uptake is defective because of a mutation in Nramp2 (now DMT1, also called DCT1), the protein responsible for endosomal iron efflux. Hence, Belgrade reticulocytes are iron deficient. We show that a chromatographic method is able to measure the amount of 'free' heme in reticulocytes. Most of the 'free' heme is the result of biosynthesis. Succinylacetone, an inhibitor of heme synthesis, decreases the level of 'free' heme and cycloheximide, an inhibitor of globin synthesis, increases the 'free' heme level. In a pulse-chase experiment with 59Fe-transferrin, the 'free' heme pool behaves as an intermediate, with a half-life of just over 2 h. Belgrade reticulocytes contain about 40% as much 'free' heme as do heterozygous or homozygous reticulocytes. This deficiency of 'free' heme slows initiation of translation in Belgrade reticulocytes by increasing the level of an inhibitor of initiation. Thus the Belgrade rat makes a whole animal model available with chronic heme deficiency.
Anemia, Hypochromic↗
A direct repeat sequence at the Rasgrf1 locus and imprinted expression.
Genomic imprinting is an epigenetic modification that can lead to parental-specific monoallelic expression of specific autosomal genes. While methylation of CpG dinucleotides is thought to be a strong candidate for this epigenetic modification, little is known about the establishment or maintenance of parental origin-specific methylation patterns. We have recently identified a portion of mouse chromosome 9 containing a paternally methylated region associated with a paternally expressed imprinted gene, Ras protein-specific guanine nucleotide-releasing factor 1 (Rasgrf1). This area of chromosome 9 also contains a short, direct tandem repeat in close proximity to a paternally methylated NotI site 30 kb upstream of Rasgrf1. Short, direct tandem repeats have been found associated with other imprinted genes and may act as important regulatory structures. Here we demonstrate that two rodent species (Mus and Rattus) contain a similar direct repeat structure associated with a region of paternal-specific methylation. In both species, the Rasgrf1 gene shows paternal-specific monoallelic expression in neonatal brain. A more divergent rodent species (Peromyscus) appears to lack a similar repeat structure based on Southern Blot analysis. Peromyscus animals show biallelic expression of Rasgrf1 in neonatal brain. These results suggest that direct repeat elements may play an important role in the imprinting process.
Animals↗
Non-transferrin-bound iron uptake in Belgrade and normal rat erythroid cells.
Belgrade (b) rats have an autosomal recessive, microcytic, hypochromic anemia. Transferrin (Tf)-dependent iron uptake is defective because of a mutation in DMT1 (Nramp2), blocking endosomal iron efflux. This experiment of nature permits the present study to address whether the mutation also affects non-Tf-bound iron (NTBI) uptake and to use NTBI uptake compared to Tf-Fe utilization to increase understanding of the phenotype of the b mutation. The distribution of 59Fe2+ into intact erythroid cells and cytosolic, stromal, heme, and nonheme fractions was different after NTBI uptake vs. Tf-Fe uptake, with the former exhibiting less iron into heme but more into stromal and nonheme fractions. Both reticulocytes and erythrocytes exhibit NTBI uptake. Only reticulocytes had heme incorporation after NTBI uptake. Properly normalized, incorporation into b/b heme was approximately 20% of +/b, a decrease similar to that for Tf-Fe utilization. NTBI uptake into heme was inhibited by bafilomycin A1, concanamycin, NH4Cl, or chloroquine, consistent with the endosomal location of the transporter; cellular uptake was uninhibited. NTBI uptake was unaffected after removal of Tf receptors by Pronase or depletion of endogenous Tf. Concentration dependence revealed that NTBI uptake into cells, cytosol, stroma, and the nonheme fraction had an apparent low affinity for iron; heme incorporation behaved like a high-affinity process, as did an expression assay for DMT1. DMT1 serves in both apparent high-affinity NTBI membrane transport and the exit of iron from the endosome during Tf delivery of iron in rat reticulocytes; the low-affinity membrane transporter, however, exhibits little dependence on DMT1.
Anemia, Hypochromic↗
Cellular distribution of iron in the brain of the Belgrade rat.
In this study, we investigated the cellular distribution of iron in the brain of Belgrade rats. These rats have a mutation in Divalent Metal Transporter 1, which has been implicated in iron transport from endosomes. The Belgrade rats have iron-positive pyramidal neurons, but these are fewer in number and less intensely stained than in controls. In the white matter, iron is normally present in patches of intensely iron-stained oligodendrocytes and myelin, but there is dramatically less iron staining in the Belgrade rat. Those oligodendrocytes that stained for iron did so strongly and were associated with blood vessels. Astrocytic iron staining was seen in the cerebral cortex for both normal rats and Belgrade rats, but the iron-stained astrocytes were less numerous in the mutants. Iron staining in tanycytes, modified astrocytes coursing from the third ventricle to the hypothalamus, was not affected in the Belgrade rat, but was affected by diet. The results of this study indicate that Divalent Metal Transporter 1 is important to iron transport in the brain. Iron is essential in the brain for basic metabolic processes such as heme formation, neurotransmitter production and ATP synthesis. Excess brain iron is associated with a number of common neurodegenerative diseases. Consequently, elucidating the mechanisms of brain iron delivery is critical for understanding the role of iron in pathological conditions.
Amino Acid Substitution↗
Nramp2 is mutated in the anemic Belgrade (b) rat: evidence of a role for Nramp2 in endosomal iron transport.
The Belgrade (b) rat has an autosomal recessively inherited, microcytic, hypochromic anemia associated with abnormal reticulocyte iron uptake and gastrointestinal iron absorption. The b reticulocyte defect appears to be failure of iron transport out of endosomes within the transferrin cycle. Aspects of this phenotype are similar to those reported for the microcytic anemia (mk) mutation in the mouse. Recently, mk has been attributed to a missense mutation in the gene encoding the putative iron transporter protein Nramp2. To investigate the possibility that Nramp2 was also mutated in the b rat, we established linkage of the phenotype to the centromeric portion of rat chromosome 7. This region exhibits synteny to the chromosomal location of Nramp2 in the mouse. A polymorphism within the rat Nramp2 gene cosegregated with the b phenotype. A glycine-to-arginine missense mutation (G185R) was present in the b Nramp2 gene, but not in the normal allele. Strikingly, this amino acid alteration is the same as that seen in the mk mouse. Functional studies of the protein encoded by the b allele of rat Nramp2 demonstrated that the mutation disrupted iron transport. These results confirm the hypothesis that Nramp2 is the protein defective in the Belgrade rat and raise the possibility that the phenotype shared by mk and b animals is unique to the G185R mutation. Furthermore, the phenotypic characteristics of these animals indicate that Nramp2 is essential both for normal intestinal iron absorption and for transport of iron out of the transferrin cycle endosome.
Alleles↗
Pollution-tolerant allele in fingernail clams (Musculium transversum).
For nearly 50 years, the fingernail clam (Musculium transversum) was believed to be virtually eliminated from the Illinois River. In 1991, workers began finding substantial populations of M. transversum in the Illinois River including several beds in and around the highly polluted Chicago Sanitary District. In order to determine if populations of M. transversum from polluted sites exhibited any genetic response to the high levels of toxins and to examine the genetic structure of several populations of M. transversum for any changes due to the population crash, starch-gel electrophoresis was performed on M. transversum from three Illinois River localities and four Mississippi River basin locations. The sampled populations produced an inbreeding coefficient (FIS) of 0.929, indicating that the populations were highly inbred. The results of a suspected founder effect due to a bottleneck was suggested by an FST = 0.442. The isozyme Glucose-6-phosphate isomerase-2 (Gpi-2) produced allelic frequency patterns that were consistent with expected patterns of a pollution-tolerant allele. Polluted sites exhibited elevated frequencies of Gpi-2(100) whereas nonpolluted sites exhibited elevated frequencies of Gpi-2(74). This frequency pattern suggested that natural selection was occurring in populations under severe toxic pressures, leading to an increase in the frequency of the allele Gpi-2(100). Therefore, Gpi-2(100) is a possible pollution-tolerant mutation in M. transversum.
Animals↗
Seven-year follow-up of coronary artery bypasses performed with and without cardiopulmonary bypass.
BACKGROUND: There has been resurgent interest in coronary revascularization performed on the beating heart. Heretofore, there has been no long-term comparison of this technique to traditional coronary artery bypass with cardioplegia. OBJECTIVE: The purpose of this study was to provide a comparison of long-term survival and intervention-free outcome between patient groups subjected to coronary bypass accomplished with or without the use of cardiopulmonary bypass. METHOD: From June 1989 to July 1990, all patients treated for coronary revascularization by three surgeons were considered for coronary revascularization with the heart beating: 107 patients underwent coronary bypass on the beating heart, and 112 patients underwent revascularization with the aid of bypass with cardioplegia. Mean ages (65 +/- 10 years) and risk factors were identical. Patients operated on with the heart beating had 2.4 +/- 0.9 grafts versus 3.2 +/- 1.1 grafts for patients having cardiopulmonary bypass with cardioplegia. RESULTS: At 7-year follow-up, 86 of 107 (80%) patients operated on with the heart beating were alive versus 88 of 112 (79%) patients in whom cardiopulmonary bypass with cardioplegia was used. Cardiac deaths occurred in 13 of 107 (12%) patients in the former group versus 10 of 112 (9%) patients in the latter group. However, 32 of 107 patients operated on with the heart beating (30%) needed catheterization for their symptoms versus 18 of 112 (16%) patients in the bypass with cardioplegia group (p = 0.01). This results in 21 of 107 (20%) patients in the beating heart group needing angioplasty or a second coronary bypass versus only 8 of 112 (7%) patients in the bypass with cardioplegia group. No patient in the bypass with cardioplegia group required reoperation. Most of the reinterventions for the beating heart group were percutaneous transluminal coronary angioplasty (15 of 21 [71%] patients). CONCLUSION: Despite one less graft per patient, survival and cardiac death rates were similar for the two groups. However, twice as many patients in the beating heart group required recatheterization (30% versus 16%), and 20% needed a second intervention. Only 7% of the bypass with cardioplegia group required reintervention. Limited revascularization of the beating heart provides long-term results comparable to full revascularization with cardiopulmonary bypass, but at the cost of a threefold increase in reinterventions.
Aged↗
The effects of salinity, pH and temperature on the half-life and longevity of Echinostoma caproni miracidia.
Miracidia of Echinostoma caproni were exposed to solutions varying in salinity, pH, and temperature in 1 ml concavity slides. Half-lives of the miracidial populations were determined and longevity curves constructed to find maximum life spans of the miracidia in the different conditions. Control miracidia in aquarium water at pH 7.2 and 22 degrees C. had a half-life of 3.6 h and a maximum life span of 9 h. Miracidia of E. caproni were not very tolerant of saline solutions from 0.1% to 0.4%, the latter being lethal within an hour. A bimodal effect was found with exposure to aquarium water of varying pH, with a peak at pH 5 in acid solutions and pH 9 in alkaline solutions. Miracidia tolerated pH ranges from 3 to 11 exhibiting half-lives of 2.4 h or greater in these solutions. At lower than ambient temperatures, E. caproni miracidia lived longer, the greatest being a half-life of 5.0 h and a maximum life span of 15 h at 5 degrees C. At warmer temperatures, the half-life was reduced until lethality was reached at 40 degrees C. ANCOVA analysis of log transformed longevity curves supported the observation that in pH trials many miracidia survived initially with a major die-off after 3.3 h. The opposite trend of early die-off and gradual mortality of the survivors was supported in the temperature trials.
Animals↗
Primary pulmonary hypertension: improved long-term effects and survival with continuous intravenous epoprostenol infusion.
OBJECTIVES: This study sought to determine the long-term effects of continuous infusion of epoprostenol (epo) therapy on survival and pulmonary artery pressure in patients with primary pulmonary hypertension (PPH). BACKGROUND: PPH is a progressive disease for which there are few effective therapies. METHODS: Patients with PPH and New York Heart Association functional class III or IV symptoms of congestive heart failure underwent right heart catheterization and Doppler-echocardiography to measure the maximal systolic pressure gradient between the right ventricle and right atrium (delta P) and cardiac output (CO). Doppler-echocardiography and catheterization data were compared. Patients were followed up long term with Doppler-echocardiography. RESULTS: Of 69 patients who went on to receive epo, 18 were followed up for > 330 days (range 330 to 700). During long-term follow-up, there was a significant reduction in delta P, which decreased from 84.1 +/- 24.1 to 62.7 +/- 18.2 (mean +/- SD, p < 0.01). A Kaplan-Meier plot of survival of our study patients demonstrated improved survival compared with that of historical control subjects. The 1-, 2- and 3-year survival rates for our patients were 80% (n = 36), 76% (n = 22) and 49% (n = 6) compared with 10- (88%, n = 31), 20- (56%, n = 27) and 30-month (47%, n = 17) survival rates in historical control subjects. CONCLUSIONS: Patients receiving continuous infusion of epo for treatment of PPH experience a decrease in pulmonary artery pressure. Long-term follow-up of this single-center patient group demonstrated improved long-term survival during epo therapy compared with that in historical control subjects and confirms predicted improved outcomes based on shorter follow-up periods.
Adolescent↗
Role of endothelin-1 in carrageenin-induced inflammation.
Many vasal factors are produced during an experimental model of inflammation such as rat-paw oedema induced by carrageenin. We investigated whether among the other well-known mediators of inflammation, i.e. serotonin, PAF, eicosanoids and kinins, the peptide endothelin-1 is produced by this kind of inflammatory process caused by carrageenin. Our results indicated that plasma endothelin, and the tissue concentration of endothelin in the oedematous paw, is increased as compared to the control. Consequently, endothelin should also be considered as an important factor in inflammatory processes.
Animals↗