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R Adar

Publications and source records attributed to R Adar.

At least 19 recordsLinked to original sources
PubMed1992

Cloning, sequence analysis and expression in Escherichia coli of the cDNA encoding a precursor of peanut agglutinin.

The cDNA coding for pre-peanut agglutinin (PNA) was isolated from a bacterial expression library. It codes for a polypeptide of 273 amino acids composed of a hydrophobic signal peptide of 23 amino acids and a mature protein of 250 amino acids. The sequence of the latter is identical to that of native PNA, determined very recently by conventional methods, except that it contains 14 additional amino acids at the C-terminus. Bacterial cells harboring a plasmid with the prePNA-cDNA, produced two PNA cross-reacting proteins: one migrated on SDS-PAGE identically with the native lectin (apparent mol. wt. 31 kDa); the other, at 35 kDa, was a beta-galactosidase pre-PNA fusion protein. The former protein possessed an N-terminal sequence identical to that of the mature, native PNA, suggesting that it was processed from the 35 kDa prePNA precursor. Only the 31 kDa protein was exported into the bacterial periplasmic space, and had the ability to bind to galactose-Sepharose. The isolated processed protein had the same hemagglutinating activity as the native lectin, when assayed with sialidase-treated human erythrocytes. Like the native lectin, it did not agglutinate the untreated cells, was not inhibited by N-acetylgalactosamine, and was inhibited by Gal beta 1----3GalNAc 30-times more strongly than by galactose.

Amino Acid Sequence↗
PubMed1992

Expression of Erythrina corallodendron lectin in Escherichia coli.

The cDNA of the Erythrina corallodendron lectin (ECorL) has been expressed in Escherichia coli. For this purpose, an NcoI site was inserted into the cDNA coding for the lectin precursor [Arango, R., Rozenblatt, S. & Sharon, N. (1990) FEBS Lett. 264, 109-112] immediately before the codon GTG (103-105) which codes for the N-terminal valine of the mature lectin. This introduced an ATG codon for a methionine preceding the valine. The mutated cDNA was ligated into pUC-8, then subcloned into the expression vector pET-3d, which carries a strong promoter derived from gene 10 of the phage T7. The recombinant plasmid was introduced into the E. coli lysogenic strain BL21(DE3). Recombinant ECorL was expressed by growing the bacteria in the presence of isopropyl beta-D-thiogalactopyranoside. Most of the recombinant lectin was found in an insoluble aggregated form as inclusion bodies and only a small part was in the culture medium in a soluble active form. Functional recombinant lectin was recovered from the inclusion bodies by solubilization with 6 M urea in cyclohexylaminopropane sulfonate pH 10.5, renaturation by 10-fold dilution in the same buffer and further adjustment of the pH to 8.0. The recombinant lectin, obtained at a yield of 4-7 mg/l culture, had, by gel filtration, a slightly lower molecular mass (56 kDa) than the native lectin, and was devoid of covalently linked carbohydrate; it was, however, essentially indistinguishable from native ECorL by other criteria, including its dimeric structure, Western blot analysis with anti-ECorL polyclonal and monoclonal antibodies, and Ouchterlony double-diffusion analysis with polyclonal antibodies, as well as hemagglutinating activity and specificity for mono- or disaccharides.

Amino Acid Sequence↗
PubMed1992

Laser Doppler flowmetry in lower extremity ischemia: application and interpretation.

The present study was undertaken in order to develop numerical criteria for the parameters of laser Doppler flowmetry-skin blood flow velocity and pulse wave amplitude in the evaluation of lower extremity ischemia. Fifty limbs of young healthy volunteers were examined in order to obtain baseline normal values. Patient population was divided into moderately ischemic (52 limbs) and severely ischemic (22 limbs), based on patients' complaints, physical examination, ankle-brachial indices, pulse volume recordings and arteriographies. Univariate comparison between the three groups of skin blood flow velocity and pulse wave amplitude at each time point were done by analysis of variance. Power of discrimination between degrees of ischemia was evaluated by computing sensitivity and specificity, based on skin blood flow velocity and pulse wave amplitude values at time points best for predicting the severity of disease by multiple regression analysis. Using cutoff points of < 31 for skin blood flow velocity and < 9 for pulse wave amplitude, a 96% sensitivity and 96% specificity were obtained in separating normal from ischemic limbs. Cutoff points of < 9 for skin blood flow velocity and < 4 for pulse wave amplitude discriminated with a 100% sensitivity and specificity between the moderately and severely ischemic limbs. We propose the use of laser Doppler flowmetry with local heating and reactive hyperemia, and the cutoff points stated above as an additional tool to evaluate lower extremity ischemia through assessment of cutaneous microcirculation.

Adult↗
PubMed1992

Autoimmune mechanisms in thromboangiitis obliterans (Buerger's disease): the role of tobacco antigen and the major histocompatibility complex.

This study is a continuation of our previous work that showed that patients with thromboangiitis obliterans (TAO; Buerger's disease) demonstrate a cell-mediated immune response to human artery type-specific collagens. To investigate the role of cigarette smoking in patients with TAO, cellular and humoral sensitivity was tested to a tobacco glycoprotein (TGP) antigen in 13 patients with Buerger's disease, 16 healthy smokers, and 12 nonsmoking healthy young male subjects. In this study, patients with Buerger's disease and healthy smokers had the same rate of cellular response to TGP, whereas nonsmokers did not respond. All three groups had a 30% to 40% measurable antibody response to TGP. If TGP has an immunologic role in the pathogenesis of TAO, an additional factor (or factors) may be operative. A specific genetic makeup may be one such factor, although at this stage other pathogenic mechanisms cannot be ruled out. Eleven patients with Buerger's disease and two control groups of 10 young healthy smoking male subjects and 12 young nonsmokers underwent histocompatibility leukocyte antigen (HLA) typing. Patients with Buerger's disease had a statistically significantly higher frequency of HLA-DR4 and a significantly lower frequency of the HLA-DRW6 antigen than had both control groups. Because similar findings have been reported in other autoimmune diseases, this observation may serve as further evidence that an autoimmune mechanism is involved in Buerger's disease.

Antibody Formation↗
PubMed1991

Changes in plasmatic tissue-type plasminogen activator and plasminogen activator inhibitor activity during acute arterial occlusion associated with severe ischemia.

Severe lower limb ischemia in patients with acute arterial occlusion was associated with a significant increase in systemic fibrinolytic activity. Plasmatic level of t-PA activity was twice the normal value at the peak of ischemia. This level declined gradually within no less than 40 hours after reperfusion procedure or limb amputation had reverted the ischemic state. In spite of major tissue damage and surgical trauma, plasmatic PAI activity stayed within normal range, and did not increase within the first 24 hours postoperatively. These findings strongly suggest that acute ischemia initiates systemic induction of excessive and continuous release of t-PA, which outweighs any anticipated increase in PAI activity.

Arterial Occlusive Diseases↗
PubMed1991

Vascular insufficiency quantitatively aggravates diabetic neuropathy.

The effect of lower-limb ischemia on the severity of neuropathy was examined in 48 diabetic patients with peripheral vascular disease. The severity of the vascular disease, as determined by medical history, physical findings, and laboratory data, was scored for each leg. Neuropathy was rated clinically and based on the results of nerve conduction studies of the common peroneal, posterior tibial, and sural nerves. A significant correlation was found between the vascular scores and neurologic variables of the two legs, most strikingly so in electrophysiologic data, with coefficients of .6 to .7. Nondiabetic control patients showed no evidence of neuropathy, regardless of the severity of ischemia, whereas diabetic controls without limb ischemia showed symmetrical neuropathy. These findings support the hypoxic theory in the pathogenesis of diabetic neuropathy.

Adult↗
PubMed1991

Pulse volume recording disturbances in paraplegic patients.

Spinal cord injuries may modify vascular reactivity in the denervated region. This study presents an original observation of altered vascular response in paraplegic patients. A group of 30 consecutive paraplegic otherwise normal individuals underwent a thorough vascular examination. There were 29 male and 1 female patient, 21 to 67 years old (average 41), all with traumatic spinal cord injury. Average time since injury was 17 years. All had good peripheral pulses and normal segmental Doppler pressure measurements. In 8 patients, plethysmography--pulse volume recording (PVR) was normal as expected. In 22 patients an unusual feature of the vascular examination was recorded, consisting of normal peripheral arterial pressures with PVR waveforms indicating poor pulsatility. This group was older than the group with normal studies: ages 44 +/- 13 vs 34 +/- 8 years (p = 0.05), and more time had elapsed since injury-20 +/- 12 vs 10 +/- 4 years (p = 0.015). The altered pulsatility demonstrated in most of those paraplegic patients may play a role in deficient wound healing frequently observed below the level of spinal neurological loss.

Adult↗
PubMed1979

Saphenous neuralgia: a complication of vascular reconstructions below the inguinal ligament.

Although relatively frequent in our experience saphenous neuralgia (SN) is not usually reported as a complication of vascular operations below the inguinal ligament. In 55 patients undergoing extended deep femoral angioplasty (EDFA, n = 28) and femoropopliteal bypass graft (FPBG, n = 27) special attention was paid to incidence and severity of postoperative SN. Severe early postoperative SN was seen in 8/28 patients with EDFA and in 6/27 with FPBG. Milder SN was seen in 10 more patients with EDFA, and 3 other developed SN many months after surgery. The milder forms of SN and late SN were not encountered after FPBG. SN usually improved with the passage of time, and at last follow-up averaging 18 months for EDFA and 33 months for FPBG there remained only 23 patients with mild SN (15 after EDFA and 8 after FPBG). The etiology of SN appears to be trauma to the nerve sustained during operation. Age, sex, diabetes, or the addition of lumbar sympathectomy to the vascular operation did not affect the risk of sustaining early postoperative SN. Increased awareness of this complication may help to understand its pathophysiology better, and possibly to decrease its incidence.

Adult↗