Functionally distinct high and low molecular weight species of channel catfish and mouse IL-1

Culture supernatants from channel catfish monocytes exhibit IL-1-like activity for mouse and catfish T cells. Gel filtration analyses of these supernatants indicated that there were at least two forms of IL-1-like activity, i.e. a high molecular weight form (70 kD) active on channel catfish, but not mouse, T cells and a low molecular weight form (approximately 15 kD) with activity for mouse, but not catfish, T cells. Both sizes of catfish IL-1 exhibited alpha and beta determinants as shown by Western blot analyses using antisera to human IL-1 alpha and IL-1 beta. Further evidence for the IL-1 nature of these molecules was obtained by antibody inhibition assays wherein antisera to human IL-1 alpha and IL-1 beta each neutralized approximately 50% of the catfish activities, were additive to some extent, and could be reversed by the addition of the proper human recombinant protein. In culture supernatants of murine P388D1 cells functional activities for catfish and mouse T cells were found only in high and low molecular weight fractions, respectively. Western blots with antiserum to mouse IL-1 alpha revealed IL-1 determinants in both high and low molecular fractions of the mouse cell culture supernatants. These data suggest that catfish and mammalian IL-1 molecules may be quite similar with the caveat being that functional activity for catfish T cells requires a large protein, presented as an aggregate, a polymer, or simply a single chain 70 kD protein. However, only the low molecular weight forms (30 and 15 kD) are active on mouse T cells.     

Comparison of biological activity of high molecular weight and low molecular weight forms of immunoreactive prolactin from human serum in cultured lymphoma NB2 cells

The aim of this study was to determine the cardiac performance of conscious healthy dogs during stimulation with dobutamine. Eight healthy unsedated beagle dogs were used. Cardiac output was measured by the thermodilution technique and blood pressures by extravascular pressure transducers. Dobutamine challenge at a dosage ranging from 27.5 to 50 micrograms kg-1 min-1 induced a significant rise in cardiac power index (CPI), cardiac index (CI), stroke index (SI) and heart rate (HR) and a significant decrease in pulmonary vascular resistance (PVR) and systemic vascular resistance (SVR). The highest CPI was 2.05 times greater than its basal resting value. The CI was primarily responsible for this increase in CPI. The SI and HR contributed approximately 55 per cent and 45 per cent respectively of the maximal increase in CI.     

Insulin increases the daily food intake of diabetic rats on high and low fat diets

The effects of insulin dose and diet composition on daily food intake were investigated by IV infusion of insulin in doses of 2 to 5 U/day into diabetic rats consuming either a high CHO or high fat diet. The daily food intake of the diabetic rats on both diets increased significantly over baseline levels (p < .01) at the low insulin doses and was maintained at these elevated levels through the 5 U/day dose. Insulin increased the rate of weight gain from Ig/day during baseline to 2 and 2.5 g/day in high CHO and high fat fed diabetics (p < .01). These results show that treatment of diabetic rats with continuous low doses of IV insulin results in a 40% increase in daily food intake regardless of the diet consumed and this increase is accompanied by an increase in rate of body weight gain. While the high fat fed diabetics were relatively hypoglycemic, these increases in intake are not the result of insulin-induced hypoglycemia, since blood glucose concentrations are significantly elevated when the increases occur at the lower insulin doses (p < .01). Thus, peripheralinsulin infused at physiological levels stimulates rather than inhibits daily food intake.     

Comparison of the in vitro fibrinolytic activities of low and high molecular weight single-chain urokinase-type plasminogen activator

The fibrinolytic activity of low molecular weight (LMW) single-chain urokinase-type plasminogen activator (scu-PA) lacking the epidermal growth factor domain and the kringle domain was compared with the activity of high molecular weight (HMW) scu-PA. LMW scu-PA was 1-5 times less active than HMW scu-PA in a fibrin plate method, in a purified fibrin clot lysis assay and in a plasma clot lysis assay. Time course experiments in a chromogenic plasminogen activator assay suggested that LMW scu-PA was less sensitive to activation by plasmin than HMW scu-PA. This was confirmed in a scu-PA activation test, which showed that at a concentration of 40 IU/ml LMW scu-PA required a three-fold higher plasmin concentration for 50% activation in 20 min than did HMW scu-PA. Kinetic experiments in the presence of 0.1 M NaCl showed non-standard Michaelis-Menten kinetics for the activation by plasmin of both HMW and LMW scu-PA. In contrast, standard kinetics was observed at 0.15 M NaCl, showing a 2.6-fold lower catalytic efficiency for LMW scu-PA than for HMW scu-PA. It is concluded that the plasmin activation of LMW scu-PA is about three times slower than the activation of HMW scu-PA. This explains, at least partially, the lower fibrinolytic activity of LMW scu-PA in comparison with HMW scu-PA.     

A new method to measure iron absorption from the enrichment of 57Fe and 58Fe in young erythroid cells

Iron absorption can be measured by the incorporation of stable iron isotopes into erythrocytes, 14 days after isotope administration. The disadvantage of this method is the high dose of isotopes needed to obtain a sufficient enrichment. Therefore, in this study cell fractions rich in young erythroid cells were prepared by using a density separation method. From 10 women blood was taken 4, 5, and 7 days after oral and intravenous administration of 57Fe and 58Fe. In these cell fractions and in whole blood taken 14 days after isotope administration, isotope enrichment was measured and absorption calculated. Absorption calculated from the isotope enrichment in the reticulocyte-rich cell fractions (12.2 +/- SEM 3.7%) was not significantly different from absorption based on whole-blood values (13.0 +/- 3.3%). Because a threefold higher isotope enrichment was found in the cell fractions, the required dose of stable isotopes can be reduced to one-third of the dose used in the traditional method without loss of sensitivity.     

Anticoagulation in hemodialysis with a single dose of low molecular weight heparin

The aim of this work was to compare the benefits and problems of low molecular weight heparin use in chronic hemodialysis, compared to conventional heparin. We studied 35 patients that received low molecular weight heparin (Enoxaparine, molecular weight 4000) during 115 consecutive hemodialysis procedures and conventional heparin during the subsequent 35 procedures. We assess the heparin dose, partial thromboplastin time before dialysis and at 3 and 120 min during the procedure, arterio-venous fistula compression time, clot formation in the circuit and residual volume of filters. Median total dose of conventional heparin was 6289 U (range 3000-10000) compared to 5555 U (range 2000-8000) of low molecular weight heparin. When the dose was calculated per kg of body weight, it was lower for low molecular weight heparin than for conventional heparin (87.8 U (range 33-100) vs 100 U (range 50-176)). Partial thromboplastin time achieved was lower with low molecular weight heparin, compared with conventional heparin, at 3 (64.26 vs 125.2 sec) and 120 min (39.1 vs 84.45 sec). Clot formation, arteriovenous fistula compression time and residual volume of filters were similar for both types of heparin. It is concluded that a single dose of low molecular weight heparin simplifies anticoagulation during hemodialysis, modifies less the partial thromboplastin time and does not alter filter re-utilization.     

Presence of polyphosphate of low molecular weight in zygomycetes

Polyphosphate of average chain length corresponding to 10 phosphate units was detected in the mycelial extract of zygomycetes. Gel electrophoresis techniques commonly used for the separation and characterization of acidic mucopolysaccharides were successfully used for the detection, purification, and characterization of the polyphosphate.     

Fractional clearance of high molecular weight proteins in conscious rats using a continuous infusion method

BACKGROUND: The purported existence of "large pores" in the glomerular capillary wall has been derived primarily from studies using dextrans and Ficolls. Systematic studies using high molecular weight proteins have not been performed. One of the difficulties is that recent studies have demonstrated that albumin and other proteins undergo degradation during renal passage. Our study took into account this renal degradation in measuring the fractional clearance of various high molecular weight proteins (the hydrodynamic radii range was between 48 to 70 A). METHODS: Fractional clearances of tritium-labeled proteins were measured using ALZET osmotic pumps, which are designed to release a slow continuous infusion of tracer. Blood and urine collections were taken at 24-hour intervals over seven days and were counted for radioactivity, and glomerular filtration rate was measured by a creatinine assay. RESULTS: Steady-state levels of [3H]protein in plasma were obtained by day 6. The [3H]proteins in the plasma showed no degradation. The fractional clearances (mean +/- sd, N = 5) of the various proteins were albumin (radius = 36 A; 0.0023 +/- 0.0009), transferrin (48 A; 0.0046 +/- 0.0007), lactoperoxidase (58 A; 0. 0045 +/- 0.0005), immunoglobulin G (62 A; 0.0043 +/- 0.0009), lactate dehydrogenase (64 A; 0.0041 +/- 0.0009), and glucose oxidase (70 A; 0.0036 +/- 0.0011). CONCLUSIONS: These values suggest a weak dependence of fractional clearance on size-selective filtration, except for albumin, which undergoes a specific type of postglomerular processing. The fractional clearances were higher than expected from previous data on dextrans and Ficolls of equivalent hydrodynamic radius, and thus demonstrate that "large pores" may already exist in normal glomerular capillary walls.     

Efficacy and safety of low molecular weight heparin in renal transplantation

BACKGROUND: Deep venous thrombosis (DVT) is a common problem with potentially devastating results in patients undergoing major surgical procedures. Certain renal transplant recipients are particularly at risk for allograft loss as a consequence of renal vein and artery thrombosis. Over the past few years, low molecular weight heparin has been well established as an accepted modality of treatment and prophylaxis of DVT. The efficacy and safety of low molecular weight heparin in the prophylaxis of DVT following renal transplantation in adults has not previously been reported. METHODS: Dalteparin was administered to 120 adult renal transplant recipients postoperatively at the Oregon Health Sciences University. RESULTS: No patient developed allograft arterial or venous thrombosis. One patient developed subclavian vein thrombosis. No bleeding complications were encountered, and side effects were very minimal. CONCLUSION: Prophylaxis with dalteparin is an effective and safe modality for the prevention of thrombosis in adult patients undergoing renal transplantation.     

Extraction of rare earths and yttrium with high molecular weight carboxylic acids

The extraction behaviour of trivalent rare earths namely La, Ce, Pr, Nd, Sm, Gd, Dy and Ho including Y (M(III), where M represents rare earths and yttrium ) from chloride medium has been studied with the solutions of high molecular weight carboxylic acids such as cekanoic, naphthenic, neo-heptanoic and Versatic 10 in dodecane. The effects of equilibrium pH, extractant concentration, metal ion concentration etc have been investigated. Using slope analysis technique it has been inferred that the metal ions form monomeric complex of the type [M(HA₂)₃] with carboxylic acids (H₂A₂, the dimer form). The stoichiometry of the species has also been confirmed using non-linear least square regression method. The carboxylic acids show different behaviour for Y extraction, it resembles to that of heavy rare earth (Ho), for sterically hindered acids (neo-heptanoic and Versatic 10) and to that with lighter rare earths (Ce, Pr) for the less sterically hindered acids (cekanoic and napthenic). The extraction order for the rare earths has been found to be the same with the four acids, i.e., La < Ce < Pr < Nd < Sm < Gd < Dy < Ho. The extraction constant (K_ex) of the systems and the separation factor amongst the rare earth pairs have been evaluated.     

Posttreatment with low molecular weight heparin reduces brain edema and infarct volume in rats subjected to thrombotic middle cerebral artery occlusion

Low molecular weight heparin (LMWH) has similar efficacy to unfractionated heparin with less hemorrhagic complications. We studied the neuroprotective effect of LMWH on a rat model of focal-ischemia. Our results revealed that treatment with LMWH at 1 and 3 h following thrombotic MCA occlusion reduced brain edema and infarct size and improved clinical outcome. Treatment with LMWH initiated at 6 h after thrombin injection only partially ameliorated brain damage.     

Characterization of nephropathy induced by immunization with high molecular weight dextran

BACKGROUND: Injection of DEAE dextran into Lewis rats can produce proteinuria and has been reported as a model of IgA nephropathy. METHODS: Cationic diethyl aminoethyl (DEAE) dextran of molecular weight 500 kDa was injected into male Lewis rats. After a pre-immunization period of 3 weeks, the animals were divided into two groups: group 1 (n = 14) received daily i.v, injections of 3.5 mg of antigen, group 2 (n = 14) was injected with 1.5 mg three times per week for a total period of 6 weeks. I.v. treatment was initiated with gradually increasing doses of DEAE dextran in both groups for 1 week, after which the maintenance dose was reached. RESULTS: We observed the appearance of proteinuria in a nephrotic range after 5 weeks of i.v. injections in group 1 (urinary excretion: 332 +/- 83 mg/24 h, controls: 53 +/- 14 mg/24 h). In group 2, the proteinuria was almost equal to protein excretion of healthy rats of the same weight (67 +/- 20 mg/24 h). The serum and urine creatinine were normal. By light microscopy of kidney biopsies, the presence of focal and segmental proliferation of mesangial cells after 6 weeks of i.v. injections was identified. Immunohistochemistry revealed no deposition of IgA, IgM, IgG, or C3. Using anti-ED1 antibodies, there was no evidence of interstitial infiltration of monocytes/macrophages after 6 weeks of i.v. injections. Staining for proliferating cell nuclear antigen (PCNA) did not show the presence of proliferating cells either in glomeruli or in the interstitium. Staining with FITC-WGA lectin revealed focal and segmental loss of the negative charge in the capillary wall. By electron microscopy there was deposition of dextran in the basal membrane and segmental and focal damage of the podocyte foot processes. As the chemokine RANTES may be involved in glomerular injury, we examined the kidneys of proteinuric and non-proteinuric rats for the presence of RANTES. By indirect immunofluorescence only the proteinuric rats showed RANTES deposition in the mesangium. CONCLUSIONS: Injection of rats with DEAE dextran leads to dose-dependent proteinuria without deposition of immune complexes but with podocyte damage. This is associated with local expression of the chemokine RANTES which may play a role in proteinuria of glomerular disease.     

Fractional clearances of low molecular weight proteins in lead workers

The phenomenon of wanting to help another is a universal human experience. Individuals reach out to others in order to fulfill personal commitments, to assist loved ones, or to meet professional obligations. The purpose of this research using Parse's method was to generate the structure of this experience as lived by nurses. Findings indicate that the lived experience of wanting to help another is directing nurturing intentions amidst uplifting affirmations with others while dissonant constraints unfold new possibilities. Findings are discussed in light of Parse's theory in order to expand the knowledge base of nursing. Similarities and differences with research findings on the experiences of nurturance and caring are presented. Implications for practice and further research are specified for nurses encountering persons who express their desire to help another.     

Mutations in CLCN5 chloride channel in Japanese patients with low molecular weight proteinuria

It was hypothesized based on previous laboratory observations and theoretical concepts that astronauts working at the "Mir" station under zero gravity conditions exhibit enhanced sensitivity to geomagnetic disturbances. The result of studies made it possible to reveal the dynamics of changes in cardiovascular characteristics separately in the main phase and the phase of storm restoration. A similar dynamics of cardiac activity was observed in experimental rabbits during geomagnetic storms.     

Social differentiation of increases in low birth weight in Poland--biological consequence of recession?

The reasons of the increase in the LBW rate in Poland from 8.1% in 1985 to 8.4% in 1990 was investigated using the individual records from the birth certificates. There was differential increase of the low birth weight among the social groups. The highest increase was observed among the less educated mothers, especially in big cities. The LBW among the newborns delivered by mothers who had finished their education at the primary school level changed from 10.6% (in big cities from 14.7%) in 1985 to 12.5% (in big cities to 16.2%) in 1990.     

Oral delivery of low molecular weight heparin in rat cardiac allografts

A 44 yr-old female with severe pulmonary emphysema and reduced alpha-1-protease inhibitor (alpha1-PI) serum levels developed an acute anaphylactic reaction following the third intravenous infusion of human alpha1-PI which was administered to prevent the progression of pulmonary emphysema. Specific immunoglobulin E-antibodies against human alpha1-PI could be demonstrated in the patient's serum using an enzyme allergosorbent test. Because of the risk of further severe anaphylactic reaction, the replacement therapy with alpha1-PI was discontinued. Physicians should be aware of this rare complication.     

Evidence for the presence of a kininogen-like species in a case of total deficiency of low and high molecular weight kininogens

EBI1-ligand chemokine (ELC) is a CC chemokine constitutively expressed in various lymphoid tissues and a high-affinity functional ligand for EBI1/CCR7, a seven transmembrane G-protein-coupled receptor originally identified as an Epstein-Barr virus (EBV)-inducible gene. Here we examined chemotactic activity of ELC on peripheral blood leukocytes. ELC attracted both CD4+ and CD8+ T cells, particularly efficiently after activation with IL-2 or with phytohemagglutinin (PHA) plus IL-2, as well as CD19+ B cells, but not CD16+ NK cells, CD14+ monocytes or neutrophils. Among CD3+ T cells, ELC attracted both CD45RO- naive and CD45RO+ memory subsets. ELC also induced vigorous calcium mobilization in T cells stimulated with IL-2 with an ED50 of 3 nM. ELC fused with the secreted form of alkaline phosphatase (ELC-SEAP) specifically bound to lymphocytes and this binding was blocked only by ELC among 10 CC chemokines so far tested. Notably, lymphocytes stimulated with IL-2 or T cells expanded by PHA plus IL-2 showed much higher levels of binding than fresh lymphocytes. Consistently, CCR7 mRNA was detected in CD4+ and CD8+ T cells as well as B cells, but not in NK cells, monocytes or neutrophils, and was dramatically increased in T cells upon treatment with IL-2 or with PHA plus IL-2. Like ELC mRNA, CCR7 mRNA was expressed in various lymphoid tissues. By in situ hybridization, ELC and CCR7 mRNA were detected in the parafollicular and inner cortical regions of a lymph node, and in the parafollicular regions of an appendix. Collectively, ELC and CCR7 may be involved in the trafficking of a broad spectrum of lymphocytes, especially activated T cells, into and within various lymphoid tissues.     

Use of compact, porous units with immobilized ligands with high molecular masses in affinity chromatography and enzymatic conversion of substrates with high and low molecular masses

Different ligands with high molecular masses are immobilized on compact, porous separation units and used for affinity chromatography. In subsequent experiments different enzymes are immobilized and used for converting substrates with low and high molecular masses. Disk or tube with immobilized concanavalin A (ConA) are used as model systems for lectin affinity chromatography. The enzyme glucose oxidase is used as a standard protein to test the ConA units. Subsequently glycoproteins from plasma membranes of rat liver are separated, using units with immobilized ConA. The enzyme dipeptidyl peptidase i.v., which is used as a model protein in the experiments, is enriched about 40-fold in a single step, with a yield of over 90%. The results are only slightly better than those obtained with ConA when it is immobilized on bulk supports. The important improvement lies in the reduction of separation time to only 1 h. Experiments concerning the isolation of monoclonal antibodies against clotting factor VIII (FVIII) are carried out on disks, combining anion-exchange chromatography and protein A affinity chromatography as a model for multidimensional chromatography. Both IgG (bound to the protein A disk) and accompanying proteins (bound to the anion-exchange disk) from mouse ascites fluid are retarded and eluted separately. With the immobilized enzymes invertase and glucose oxidase (GOX) the corresponding substrates with low molecular masses, saccharose and glucose, are converted. It is shown that the amount of immobilized enzyme and the concentration of the substrate are responsible for the extent of the conversion, whereas the flow-rates used in the experiments have no effect at all. The influence of immobilization chemistry was investigated with GOX. Indirect immobilization with ConA as spacer proved to be the best alternative. With trypsin, immobilized on a disk, substrates with high molecular masses are digested in flow-through. For optimal digestion the proteins have to be denatured in the buffer for sodium dodecyl sulfate-polyacrlyamide gel electrophoresis prior to application. In contrast to the conversion of substrates with low molecular masses, flow-rates play an important part in conversion of substrates with high molecular masses. With lower flow-rates a higher degree of digestion is achieved.     

Association of factor XI and high molecular weight kininogen in human plasma

Factor XI and high molecular weight kininogen were found associated in normal human plasma at mol wt 380,000 as assessed by gel filtration on Sephadex G-200. The molecular weight of Factor XI in high molecular weight kininogen-deficient plasma was 175,000, the same value obtained for purified Factor XI. When high molecular weight kininogen-deficient plasma was reconstituted with purified high molecular weight kininogen, all of the Factor XI was then found at mol wt 380,000. Complex formation was also demonstrable upon incubation of Factor XI and highly purified high molecular weight kininogen. This complex was distinct from the prekallikrein-high molecular weight kininogen complex; thus high molecular weight kininogen forms bimolecular complexes with either Factor XI or prekallikrein but does not form a trimolecular complex that includes both Factor XI and prekallikrein. Neither Hageman factor nor plasminogen were found associated with high molecular weight kininogen; binding to high molecular weight kininogen appeared to be a specific property of the Hageman factor substrates.