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M Green

Publications and source records attributed to M Green.

At least 757 records · Page 42Linked to original sources

Transforming region of group A, B, and C adenoviruses: DNA homology studies with twenty-nine human adenovirus serotypes.

The 31 human adenovirus (Ad) serotypes form five groups based upon DNA genome homologies: group A (Ad12, 18, 31), group B (Ad3, 7, 11, 14, 16, 21), group C (Ad1, 2, 5, 6), group D (Ad8, 9, 10, 13, 15, 17, 19, 20, 22-30), and group E (Ad4) (M. Green, J. Mackey, W. Wold, and P. Rigden, Virology, in press). Group A Ads are highly oncogenic in newborn hamsters, group B Ads are weakly oncogenic, and other Ads are nononcogenic. However, most or all Ads transform cultured cells. We have studied the homology of Ad5, Ad7, and Ad12 transforming restriction endonuclease DNA fragments with DNAs of 29 Ad types. Ad5 HindIII-G (map position 0-7.3), Ad7 XhoI-C (map position 0-10.8), and Ad12 (strain Huie) EcoRI-C (map position 0-16) and SalI-C (map position 0-10.6) fragments were purified, labeled in vitro (nick translation), and annealed with DNAs of Ad1 to Ad16, Ad18 to Ad24, and Ad26 to Ad31. Hybrids were assayed by using hydroxylapatite. Ad5 HindIII-G hybridized 98 to 100% with DNAs of group C Ads, but only 1 to 15% with DNAs of other types. Ad7 XhoI-C fragment hybridized 85 to 99% with DNAs of group B Ads, but only 6 to 21% with DNAs of other types. Ad12 (Huie) EcoRI-C hybridized 53 to 68% with DNAs of five other Ad12 strains, 53% with Ad18 DNA, 56% with Ad31 DNA, but only 3 to 13% with DNAs of other types. In vitro-labeled Ad12 (Huie) SalI-C hybridized 35 to 71% with DNAs of 6 other Ad12 strains, 44% with Ad18 DNA, 52% with Ad31 DNA, but only 2 to 7% with DNAs Ad7, Ad2, Ad26, or Ad4. When assayed using S-1 nuclease, SalI-C annealed 17 to 44% with DNAs of group A Ads. The melting temperatures of the hybrids of Ad5 HindIII-G with all group C Ad DNAs were 84 degrees C in 0.12 M sodium phosphate (pH 6.8). The melting temperature of the Ad12 (Huie) EcoRI-C hybrid with Ad12 (Huie) DNA was 83 degrees C, but was only 71 to 77 degrees C with DNAs of other group A Ads. Thus, group C and group B Ads both have very homologous transforming regions that are not represented in DNAs of non-group C Ads or non-group B Ads, respectively. Similarily, group A Ads have unique but less homologous transforming regions. These different transforming nucleotide sequences may be reflected in the different oncogenic properties of group A, B, and C Ads.

Adenoviruses, Human↗

Adenovirus type 2 early polypeptides immunoprecipitated by antisera to five lines of adenovirus-transformed rat cells.

We have identified adenovirus type 2 (Ad2)-induced early polypeptides (EPs) and have attempted to determine which EPs are coded by each of the four early gene blocks. [35S]methionine-labeled EPs were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cycloheximide pretreatment followed by labeling in hypertonic medium (210 to 250 mM NaCl) facilitated the detection of EPs. Seven major (reproducible bands in autoradiograms) EPs were detected with molecular weights of 74,000 (74K), 21K, 19K, 15K, 13.5K, 11.5K, and 11K. Minor (weaker bands) EPs of 55K, 52K, 42K, 18K, 12K, 8.8K, and 8.3K were also often seen. To identify and map the genes for virus-coded EPs, we prepared antisera against five lines of adenovirus-transformed cells that retain different fractions of the viral genome. The lines were F17, 8617, F4, and T2C4 transformed by Ad2 virions and 5RK (clone I) transformed by transfection with the Ad5 HsuI-G fragment (map position 0 to 8). The early gene blocks retained and expressed (in part) as RNA in these cells were as follows: 5RK(I), block 1 (70% of left 8% of genome); F17, block 1; 8617, blocks 1 and 4; F4 blocks 1, 2, and 4; T2C4, blocks 1, 2, 3, and 4. The following major EPs were immunoprecipitated: 15K by all antisera; 53K and 14.5K by F17, T2C4, 8617, and F4 antisera; 11.5K by T2C4, 8617, and F4 antisera; 44K, 42K, 19K, and 13.5K by T2C4 antisera; 11K by 8617 antisera. Minor EPs of 28K, 18K, and 12K were precipitated by all antisera except 5RK(I). The 53K and 15K EPs were precipitated also from Ad2 early infected monkey cells by the F17 antiserum and by sera from hamsters bearing tumors induced by Ad1-simian virus 40. The relationships between some of the immunoprecipitated EPs were investigated by the partial proteolysis procedure. All 53K EPs are the "same" (i.e., highly related), all 15K EPs are the "same," and all 11.5K EPs are the "same." The 15K EP is highly related to the 14.5 K EP. Although less certain, all 28K EPs appeared related, as did all 18K EPs. The T2C4-specific 44K EP is probably a dimer of the 21K glycopolypeptide. The T2C4-specific 13.5K EP and the 8617-specific 11K EP appear unrelated to any other polypeptides. These immunoprecipitation data provide evidence that early gene block I (map position 1 to 11) may encode major 53K, 15K, and 14.5K polypeptides, and minor 28K, 18K, and 12K polypeptides, and that all or some of the gene for 15K and 14.5K lies within map position 1 to 8. The surprisingly complex pattern of polypeptides coded by early gene block I raises the possibility that some polypeptides may be coded by overlapping "spliced" mRNA's. The possible block locations of the genes for the 21K, 13.5K, and 11.5K polypeptides are discussed.

Adenoviruses, Human↗

Protein synthesized early after infection is linked to the termini of adenovirus type 2 DNA synthesized in vivo and in vitro.

The human adenovirus DNA genome contains a protein (CBP, or covalently bound protein) linked to each 5' terminus. To assess whether CBP is synthesized early, infected cells were incubated with hydroxyurea from 1 to 18 h postinfection, the hydroxyurea was removed, cycloheximide was added, and viral DNA was labeled with [3H]thymidine from 18 to 23 h postinfection. Removal of hydroxyurea at 18 h postinfection permits the synthesis of viral DNA, whereas cycloheximide maintains the block in late viral protein synthesis. Three lines of evidence are presented to show that viral 3H-labeled DNA prepared by this procedure was linked to CBP: (I) the DNA sedimented more rapidly than protein-free DNA (i.e., protinase treated) in neutral sucrose gradients containing guanidine hydrochloride; (ii) the DNA banded at a lower density than protein-free DNA in CsCl gradients containing guanidine hydrochloride; and (iii) neither the 3H-labeled DNA nor the end fragments produced by EcoRI digestion entered a 1.4% agarose gel during electrophoresis. These experiments are strong evidence that CBP is not a product of a late viral gene and is therefore the product of either an early viral gene or a cell gene. Experiments were performed to test whether CBP is attached to viral DNA synthesized in vitro by a soluble complex that synthesizes exclusively viral DNA as completed viral genomes in vitro. In vitro-labeled DNA was analyzed by velocity sedimentation, equilibrium sedimentation, and agarose gel electrophoresis as described above. Our results indicate that the majority of in vitro-synthesized DNA molecules were attached to CBP. These results, which indicate that CBP is synthesized early after infection and is attached to viral DNA labeled in vitro by a soluble replication complex, are consistent with the idea that CBP may play a role in viral DNA replication.

Adenoviruses, Human↗

The 55K protein on the 5' termini of adenovirus type 2 DNA is unrelated to virus-coded candidate transformation proteins (E1-53K, E1-40K-50K) and DNA-binding proteins (E2-42K/47K/73K).

A polypeptide of 55,000 daltons (55K) is linked, probably covalently, to the K' termini of adenovirus type 2 DNA. The 55K polypeptide is synthesized during early stages of infection (T. Yamashita, M. Arens, and M. Green, J. Virol. 30: 497-507, 1979) and thus may function in viral DNA replication, gene regulation, or cell transformation. Several virus-coded early polypeptides have been identified that could correspond to the terminal 55K, including the E1-40K-50K and E1-53K candidate transformation polypeptides and the E2-42K/47K/73K single-stranded DNA-binding polypeptide. We show here that two-dimensional tryptic [35S]methionine-peptide maps of the terminal 55K differ completely from [35S]methionine-peptide maps of four related E1-40K-50K polypeptides, the E1-53K, and the related E2-42K, E2-47K, and E2-73K polypeptides. We conclude that the terminal 55K polypeptide does not correspond to any of the known virus-coded early polypeptides.

Adenoviruses, Human↗

Occupational asthma in an electronics factory.

Workers in a modern electronics factory were surveyed by questionnaire and lung function testing to see if there was evidence of widespread work-related respiratory symptoms. Of the responding workers exposed to solder flux fumes on the shop floor, 22% had work-related breathlessness or wheeze or both. Exposed workers had a lower FEV1 and FVC than unexposed workers. Work-related rhinitis was also present in 22% of exposed workers. The most likely cause for these results is sensitivity to colophony fumes, released from solder flux during soldering. Levels of solder flux fume were below the threshold limit value in this factory during the survey.

Adult↗

Work-related respiratory disease in employees leaving an electronics factory.

Examination of the records of employees leaving an electronics factory over three-and-a-half years showed that a significantly greater proportion left the shop floor (where soldering took place) because of ill health than left the stores and office areas. This difference was largely due to work-related respiratory disease in those whose job was soldering. Shop floor workers leaving for health reasons also had increased sickness certification due to respiratory illness compared to stores and office workers. These findings suggest that work-related respiratory illness is a significant cause of morbidity and loss of employment in solderers working at the factory and that this has been a longstanding problem with its onset before the first recorded cases of occupational asthma caused by solder flux containing colophony.

Adolescent↗

Thoracoabdominal mechanics during relaxed and forced vital capacity.

Thoracoabdominal configuration, intrathoracic (esophageal), intra-abdominal (gastric), and transdiaphragmatic pressures were studied in six normal upright subjects during relaxed (RVC) and forced vital capacity (FVC). Chest wall configuration showed substantial departure from its relaxation characteristics during FVC. Paradoxical (outward) movement was recorded for a low lateral diameter of the rib cage (on the costal margin) at high volume during RVC and during most of FVC, while the other rib cage dimensions were decreasing. Transdiaphragmatic pressure was positive during most of the FVC, particularly toward RV, reflecting active contraction of the diaphragm. We conclude that diaphragmatic activity modulates forced expiration and that the chest wall may influence the FVC maneuver.

Abdomen↗

Plasma cholesterol esterifying activity in guinea pigs.

Plasma cholesterol esterifying activity has been measured in guinea pigs fed either a control diet or the same diet supplemented with 1% cholesterol. The extent of esterification was found to be similar in the cholesterol-fed and control guinea pigs and somewhat lower than in rats. The initial rate of esterification was also of the same magnitude as that found in rats and humans, and unaffected by dietary cholesterol if autologous plasma was used as substrate. However, LCAT activity from cholesterol-fed guinea pigs was significantly higher than that of control plasma when acting on either control or cholesterol-fed substrate. This suggests that dietary cholesterol increases the amount (or activity) of LCAT but that the substrate is unsuitable or that a necessary cofactor is present in limiting amounts. Heat treatment of guinea pig plasma seems to alter substrate availability to varying degrees. The implications of these findings in relation to substrate specificity and cofactor requirements of guinea pig LCAT are discussed.

Animals↗

Comparison of yes-no and latency measures of auditory intensity discrimination.

Rats discriminated auditory intensity differences of sinusoids at 3.0 kilohertz in a go/no-go signal detection procedure. Responses to the signal (hits) were reinforced with electrical brain stimulation, and misses produced a brief timeout. On intermixed noise trials, withholding of responses (correct rejections) was reinforced, and false alarms produced the time-out. In two test conditions, the signal was either the louder (100 decibels) or softer (90, 93, 96, or 99 decibels) of the pair of intensities presented within a set of trials. Each animal was first trained with signal value louder or softer, and reversed for the second condition so that the former noise value served as signal. Hits showed shorter latencies than false alarms, regardless of the relative intensity of signal and noise, and the magnitude of differentiation was proportional to signal-noise separation. Both hits and false alarms showed longer latencies as the discrimination became more difficult. Isosensitivity contours derived from the latencies showed close similarity across conditions; in comparison, the yes-no measure of detectability, d', showed greater variability. The similarity of latency differentiation across louder and softer signal conditions supports a detection model in which the observer's judgment is controlled by the distance of sensory effect from criterion on each trial, as opposed to the loudness of the tones per se.

Animals↗

Outcomes of pregnancy for addicts receiving comprehensive care.

This study reports pregnancy outcomes for 105 addicted women enrolled in New York Medical College's Pregnant Addicts and Addicted Mothers Program. Three classes of variables are examined: prenatal care variables, obstetrical outcomes, and neonatal outcomes. As a first step, percentage distributions are shown for all variables within each class. With respect to the second and third classes, comparisons are made when possible to findings reported elsewhere on heroin-addicted, methadone-maintained, and drug-free populations. Zero-order correlations are then shown for the prenatal care variables versus the neonatal outcomes. It is found that the mother's number of prenatal medical visits correlates significantly with the neonate's gestational age at birth and birth weight, and that her methadone dose at time of delivery correlates significantly with the neonate's withdrawal status. The effect of the prenatal care variables on the two key neonatal outcomes--gestional age at birth and birth weight--is then examined via a stepwise regression analysis. It is found that two of the variables--gestional duration at first prenatal visit and number of prenatal visits--together account for 18.5% of the variance in birth weight and 26.9% of the variance in gestational age at birth, while maternal methadone dose has virtually no effect on these outcome measures. The findings indicate that better neonatal outcomes are associated with the mother's joining the program relatively early in pregnancy and coming in relatively often for prenatal care.

Adult↗

Analysis of human cancer DNA for DNA sequences of human adenovirus type 4.

We investigated whether human adenovirus type 4 (Ad4) might cause cancer in humans. Cell culture and animal model studies indicated that Ad4-induced human tumors should contain Ad4 DNA sequences. Thus Ad4 DNA was labeled in vitro (sp act, approximately 10(7) 3H counts/min/microgram) and hybridized in liquid phase with DNA extracted from human tumors. No viral sequences were detected in 31 squamous cell carcinomas of the lung, 5 adenocarcinomas of the lung, 4 oat cell carcinomas of the lung, 4 carcinomas of the stomach, 10 carcinomas of the colon, 3 carcinomas of the kidney, 3 carcinomas of the breast, 2 carcinomas of the ovary, and 6 Hodgkin's lymphomas. Reconstruction experiments with added Ad4 DNA indicated that the probe detected about 1 copy of 5% of the Ad4 genome per tumor cell. Therefore, these data were strong evidence (but did not prove) that none of these particular tumors were induced by Ad4.

Adenoviruses, Human↗

Procarbazine, vinblastine, and actinomycin D in stage III and IV melanoma with or without methanol-extracted residue of Bacillus Calmette-Guérin.

Patients with stage III and IV melanoma were randomly assigned to receive procarbazine (100 mg/m2, Days 1--10), vinblastine (5 mg/m2, Days 1 and 8), and actinomycin D (0.5 mg/m2, Days 1 and 8) with or without methanol-extracted residue (MER) of bacillus Calmette-Guerin (200 micrograms in five sites). In patients with measurable disease, 20% (eight of 40 patients) responded with only the combination chemotherapy while 15% (six of 39 patients) responded with the MER added. Toxicity was tolerable except for some instances of severe, gastrointestinal toxicity associated with procarbazine. MER as given in this study, failed to either increase the response rate or prolong survival.

Antineoplastic Agents↗

Hyponatraemia and mesothelioma.

Hyponatraemia was found in 18 (62%) of 29 patients with histologically proven mesothelioma. The mechanism for the development of the hyponatraemia is discussed. It is suggested that patients with mesothelioma should be monitored for electrolyte abnormalities. If hyponatraemia is found the syndrome of inappropriate secretion of antidiuretic hormone should be suspected as treatment with demethylchlortetracycline may improve the quality of life in an otherwise fatal illness.

Female↗

Analysis of human cancer DNA's for DNA sequence of human adenovirus serotypes 3, 7, 11, 14, 16, and 21 in group B1.

We have investigated whether Group B human adenoviruses (Ad) (Ad3, Ad7, Ad11, Ad14, Ad16, and Ad21), which are widespread in the human population and are tumorigenic in hamsters, may play a role in human cancer. Hybridization of Ad7-radiolabeled DNA with DNA's from an Ad7-induced primary hamster tumor and from two cell lines (5728 and Ad7 P-cell) established from Ad7-induced hamster tumors indicated multiple copies per cell of 17, 30 to 36, and 20%, respectively of the Ad7 genome. Thus, cells transformed by Group B Ads resemble cells transformed by Group C and Group A Ad's in that they retain multiple copies of a variable fraction of the viral genome. These model studies suggest that possible Group B Ad-induced human cancer cells should contain one or more copies of virus DNA per cell. Therefore, we assayed human cancer DNA's for Ad sequences, by highly sensitive "saturation-hybridization" reactions with Ad7 or Ad11 DNA (4 X 10(6) to 2.1 x 10(8) cpm/microgram). We concentrated on cancers of the respiratory and digestive systems, because these systems are the most common sites of infection by Group B Ad's. In 8 independent experiments, no Ad7 sequences were detected in DNA's from 16 normal lung tissues, 18 normal tissues of the digestive system, 34 cancers of the respiratory system, 19 cancers of the digestive system, 11 cancers of the urinary system, 5 cancers of the genital system, 3 cancers of the breast, and 6 Hodgkin's lymphomas. Reconstruction controls with added Ad7 DNA indicated that about 0.05 to 0.1 copy of Ad7 DNA per cell should be detected. Ad11 is strongly implicated as a cause of acute hemorrhagic cystitis. In two independent experiments, no Ad11 sequences were detected in DNA's from 9 carcinomas of bladder, 10 carcinomas of prostate, 24 carcinomas of kidney, 3 hypernephromas, 3 Wilms' tumors, or 2 normal kidneys. Reconstruction experiments indicated that the cancer DNA assays had a sensitivity of 0.05 to 0.1 copy of Ad11 DNA per cell. The DNA's of Group B Ad's are greater than 85% homologous by hybridization; thus, these results are applicable to all Group B serotypes. Our data provide evidence (but not formal proof) that none of the human cancers that we analyzed were induced by Group B Ad's. These tumors represent about 50% of the tumors that affect humans. The possible involvement of Group B Ad's in other less common forms of human cancers is under investigation in our laboratory.

Adenoviruses, Human↗