Inhibition of restriction endonuclease hydrolysis by phosphorothioate-containing DNA.
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Biomedical subjects
Publications and source records attributed to D B Olsen.
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Reaction of Bacillus cereus phosphonoacetaldehyde hydrolase (phosphonatase) with phosphonoacetaldehyde or acetaldehyde in the presence of NaBH4 resulted in complete loss of enzymatic activity. Treatment of phosphonatase with NaBH4 in the absence of substrate or product had no effect on catalysis. Inactivation of phosphonatase with [3H]NaBH4 and phosphonoacetaldehyde, NaBH4 and [14C]acetaldehyde, or NaBH4 and [2-3H]phosphonoacetaldehyde produced in each instance radiolabeled enzyme. The nature of the covalent modification was investigated by digesting the radiolabeled enzyme preparations with trypsin and by separating the tryptic peptides with HPLC. Analysis of the peptide fractions revealed that incorporation of the 3H- or 14C-radiolabel into the protein was reasonably selective for an amino acid residue found in a peptide fragment observed in each of the three trypsin digests. Sequence analysis of the 3H-labeled peptide fragment isolated from the digest of the [2-3H]phosphonoacetaldehyde/NaBH4-treated enzyme identified N epsilon-ethyllysine as the radiolabeled amino acid. The ability of the phosphonatase competitive inhibitor (Ki = 230 +/- 20 microM) acetonylphosphonate to protect the enzyme from phosphonoacetaldehyde/NaBH4-induced inactivation suggested that the reactive lysine residue is located in the enzyme active site. Comparison of the relative effectiveness of phosphonoacetaldehyde and acetaldehyde as phosphonatase inactivators showed that the N-ethyllysine imine that is reduced by the NaBH4 is derived from the corresponding N-(phosphonoethyl) imine. On the basis of these findings, a catalytic mechanism for for phosphonatase is proposed in which phosphonoacetaldehyde is activated for P-C bond cleavage by formation of a Schiff base with an active-site lysine. Accordingly, an N-ethyllsysine enamine rather than the high-energy acetaldehyde enolate anion is displaced from the phosphorus.
A valveless, single-orifice abdominal aortic counterpulsation device (AACD) was implanted retroperitoneally on the abdominal aorta and was pumped continuously, providing aortic diastolic augmentation for 45 +/- 4.9 days in four calves (group A). The hematocrit, lactate dehydrogenase (LDH), bilirubin, blood urea nitrogen, and creatinine were obtained before implantation and weekly thereafter. Biocompatibility data were compared to those obtained from 11 calves (group B) that received a total artificial heart (TAH) and were electively terminated 50.0 +/- 19.5 days after implantation. The hematocrit values in the first week were 20.9 +/- 11.5% and 39.8 +/- 11.5% below control values for group A and group B, respectively (p less than 0.02); in the sixth week they were 5.1 +/- 14.6% above control values (group A) and 22.6 +/- 9.0% below control values (group B) (p less than 0.05). LDH did not change in group A, while in group B it was constantly about 100% above control values. Autopsy revealed one to three infarcts 1 to 3 mm in diameter in 63% of the kidneys in group A, while in all of group B kidneys there were multiple infarcts of more than 10 mm. After the induction of left ventricular (LV) failure, the AACD decreased the LV end-diastolic pressure (EDP) by 21.2% (p less than 0.005) and the aortic (AO) EDP by 18% (p less than 0.005). It increased the endocardial viability ratio by 300% (p less than 0.0005), and the cardiac index by 66.9% (p less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)
A rare lesion, steatocystoma simplex, in a previously unreported location, the oral cavity, is described. A case report, the differential diagnosis, and a review of the literature are offered. Differentiation between this cyst and steatocystoma multiplex is important in terms of the need for clinical follow-up.
Vegetative endocarditis and thromboembolism are the two leading pathologic findings and causes of death in calves receiving total artificial hearts (TAH). Percutaneous access devices (PAD) for the pneumatic drive lines often become infected and may serve as the nidus of infection of the vegetative lesions. Observations of 80 calves living with TAHs revealed that most infections began at the dorsal surface of the PAD. Chronic skin ulceration or acute, traumatic avulsion can result in PAD separation from the skin and bacterial implantation. Evaluation of 58 necropsy specimens from calves with TAHs demonstrated improper flange positioning in 76% of the PAD and gross infection in 57%. A new implantation technique and design of PAD have resulted in better flange position and less irritation at the skin/device interface. Necropsies of 11 animals with the new PAD have shown a reduction in superficial infections.
It became necessary to perform a staged procedure by means of total artificial heart followed by cardiac transplantation in a 26-year-old, female patient. The Ellipsoidheart, as an artificial heart, fitted excellently in the chest without compromising the surrounding structures. As soon as the patient recovered with the artificial heart, a consecutive transplantation was performed. This first European clinical experience gained confidence and suggests applying more artificial hearts as a bridge towards transplantation.
The in vitro development of automatic control of the actively filled, alternately pumped, volumetrically coupled, electrohydraulic total artificial heart was the goal of this investigation. Control features under study were (a) cardiac output (CO) response to preload; (b) CO relationship to mean aortic pressure (AoP); and (c) control of balanced ventricular outputs. A modified pulmonic valve to increase backflow was used as a balancing mechanism. Hydraulic fluid pressure transducers monitored diastolic pressures, and microprocessor control of motor speed maintained in a mild suction to yield filling rate dependent on atrial pressure. Results indicated a rise in CO from 5 to 9 L/min, with a change in mean right atrial pressure (RAP) from 0 to 7 mm Hg. No significant difference in CO was found as AoP was varied from 80 to 120 mm Hg with a maximum variation of +/- 0.5 L/min on CO and +/- 1 mm Hg on RAP. Balance was maintained for bronchial flows up to 50% with mean left atrial pressure never exceeding 15 mm Hg. An alternately pumped electrohydraulic heart was automatically controlled to respond sensitively to preload changes. Afterload changes did not alter the CO response curve. Automatically controlled, balanced ventricular outputs were maintained.
There is a variety of mechanical prosthetic valves that can be applied to blood pumps, and one can be selective about those particular characteristics tailored to each blood pump and its application. These devices have become extremely reliable, particularly since, in the experience of this laboratory, the dp/dt generated in the ventricle of the pneumatically powered artificial heart is successfully lowered. Future modifications of the valves, such as the Medtronic-Hall monostrut valve and the Medtronic-Hall tubular valve, are expected.
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The 41-residue sequences of recently identified porcine corticotropin-releasing factors [Ile40]pCRF and [Asn40]pCRF were assembled on a benzhydrylamine resin support. Deprotection and cleavage from the resin were accomplished by HF treatment. The crude peptides were purified by HPLC. The homogeneity of the final materials, obtained in 0.2% and 0.4% overall yield for [Ile40]pCRF and [Asn40]pCRF respectively, was assessed after the isolation by HPLC and amino acid analysis. Both sequences of the synthetic 41-residue pCRF stimulated the release of corticotropin (ACTH) from superfused rat pituitary cells on a column, the responses being related to a log-dose of CRF in the range of 1-20 ng/ml. [Ile40]pCRF and [Asn40]pCRF also augmented the in vivo release of ACTH in rats pretreated with chlorpromazine, morphine and Nembutal. [Ile40]pCRF appeared to be equipotent to ovine CRF and about twice as active as [Asn40]pCRF. The data indicate that synthetic porcine [Ile40]pCRF and [Asn40]pCRF have high biological activity.
The development of the total artificial heart (TAH) has reached a level where it is now available for clinical applications. The TAH has demonstrated distinct advantages over other forms of mechanical circulatory assistance. As of December 1, 50 TAHs have been implanted: 5 as permanent devices, and 45 as a temporary mechanical bridge to cardiac transplantation. The use of the TAH has increased in the last several months, leading to a growing interest in defining the indications and contraindications to its use. End-stage cardiomyopathy (either idiopathic, ischemic, viral, or postpartum) has been the underlying disease in 80% of the TAH procedures to date. The TAH has also been applied in 5 cases of acute cardiac graft rejection, 2 cases of congenital heart diseases, and in one case after acute myocardial infarction. The indications for the use of the TAH in these and other potential patient groups is discussed in light of the current clinical results.
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More and more handicapped individuals may be seeking dental treatment. Treatment may include construction of special devices to help these patients lead more productive and fulfilling lives. A mouthstick has been designed that can be constructed from readily available parts. The versatility and decreased cost of the mouthstick make it readily available to these needy patients.
The abdominal aortic counterpulsation device is a round pumping chamber with a valveless opening which is implanted retroperitoneally on the abdominal aorta. The Utah driver is connected to the device through an air conduit and is synchronized on the electrocardiographic signal to provide diastolic aortic augmentation. For comparison an intraaortic balloon was also driven by the Utah driver system. The abdominal aortic counterpulsation device (stroke volume = 30, 40 and 60 ml) and the intraaortic balloon pump (balloon volume = 20 ml) were tested in dogs with acute left ventricular failure. The abdominal aortic counterpulsation device was also tested in normal animals. In acute left ventricular failure the abdominal aortic counterpulsation device at a stroke volume of 30, 40 or 60 ml decreased left ventricular end-diastolic pressure by an average of 28.56 (p less than 0.001), 39.56 (p less than 0.001) and 44.14% (p less than 0.005), respectively; aortic end-diastolic pressure by 24.11 (p less than 0.001), 26.67 (p less than 0.001) and 19.57% (p less than 0.01); and aortic systolic pressure by 18.56 (p less than 0.002), 26.0 (p less than 0.001) and 22.43% (p less than 0.005). It increased cardiac index by 27.58 (p less than 0.02), 35.59 (p less than 0.005) and 43.42% (p less than 0.001) and it provided peak aortic diastolic augmentation of 64.5 (p less than 0.001), 69.78 (p less than 0.001) and 74.43% (p less than 0.001), respectively, above the control aortic end-diastolic pressure.(ABSTRACT TRUNCATED AT 250 WORDS)
The use of conventional open-port catheters in total artificial heart (TAH) research shortens the survival time of recipients owing to sepsis and embolism caused from the catheters. Valuable data are lost when an open-port catheter clots off, and transducer position must be keyed to the level of the atria for accuracy. The need for an accurate, easily obtainable, noninvasive method for measuring atrial pressure is of obvious value. Such a method has been developed in vitro using a device that measures the stroke volume of a pneumatic ventricle. The stroke volume is obtained by quantifying the amount of air exiting a pneumatic TAH in diastole. Using this information ventricular filling rates can be calculated by the stroke volume measurement device, and these rates are correlated to measured atrial pressures. There is no need to continually adjust transducer levels in relation to the atria with this system. The data show an average percentage error of 2.4 of full scale (25 mm Hg) or 0.6 mm Hg. The method of measurement is accurate, without limitations on driving parameters. The information is available without any additional prosthetic fabrication or surgical intervention than that already needed for basic TAH implantation. This method of measuring atrial pressures now needs to be proved effective in vivo.
This article is intended to inform the reader of the current state of the art of pneumatically powered artificial hearts, including animal experiments and information available on 11 recipients of orthotopically placed devices.