Inspiratory muscle training as adjuvant therapy in obstructive sleep apnea: a randomized controlled trial.

The aim of this randomized controlled trial was to analyze the effects of an inspiratory muscle training (IMT) program on apnea and hypopnea index (AHI), inspiratory muscle strength, sleep quality, and daytime sleepiness in individuals with obstructive sleep apnea (OSA), whether or not they used continuous positive airway pressure (CPAP (+/-) therapy. The intervention group underwent IMT with a progressive resistive load of 40-70% of the maximum inspiratory pressure (PImax) for 30 breaths once a day for 12 weeks. The control group was submitted to a similar protocol, but with at a minimum load of 10 cmH2O. Changes in the AHI were the primary outcome. PImax was measured with a digital vacuometer, daytime somnolence was measured by the Epworth sleepiness scale (ESS), and the quality of sleep by the Pittsburgh Sleep Quality Index (PSQI). CPAP use was treated as a confounder and controlled by stratification resulting in 4 subgroups: IMT-/CPAP-, IMT-/CPAP+, IMT+/CPAP-, and IMT+/CPAP+. Sixty-five individuals were included in the final analysis. Significant variations were found in the 4 parameters measured throughout the study after the intervention in both CPAP- and CPAP+ participants: PImax was increased and AHI was reduced, whereas improvements were seen in both ESS and PSQI. The twelve-week IMT program increased inspiratory muscle strength, substantially reduced AHI, and had a positive impact on sleep quality and daytime sleepiness, whether or not participants were using CPAP. Our findings reinforce the role of an IMT program as an adjunct resource in OSA treatment.

Inspiratory muscle training as adjuvant therapy in obstructive sleep apnea: a randomized controlled trial

The aim of this randomized controlled trial was to analyze the effects of an inspiratory muscle training (IMT) program on apnea and hypopnea index (AHI), inspiratory muscle strength, sleep quality, and daytime sleepiness in individuals with obstructive sleep apnea (OSA), whether or not they used continuous positive airway pressure (CPAP (+/−) therapy. The intervention group underwent IMT with a progressive resistive load of 40-70% of the maximum inspiratory pressure (PImax) for 30 breaths once a day for 12 weeks. The control group was submitted to a similar protocol, but with at a minimum load of 10 cmH2O. Changes in the AHI were the primary outcome. PImax was measured with a digital vacuometer, daytime somnolence was measured by the Epworth sleepiness scale (ESS), and the quality of sleep by the Pittsburgh Sleep Quality Index (PSQI). CPAP use was treated as a confounder and controlled by stratification resulting in 4 subgroups: IMT−/CPAP−, IMT−/CPAP+, IMT+/CPAP−, and IMT+/CPAP+. Sixty-five individuals were included in the final analysis. Significant variations were found in the 4 parameters measured throughout the study after the intervention in both CPAP− and CPAP+ participants: PImax was increased and AHI was reduced, whereas improvements were seen in both ESS and PSQI. The twelve-week IMT program increased inspiratory muscle strength, substantially reduced AHI, and had a positive impact on sleep quality and daytime sleepiness, whether or not participants were using CPAP. Our findings reinforce the role of an IMT program as an adjunct resource in OSA treatment.

Out-of-hospital airway management during manual compression or automated chest compression devices : A registry-based analysis.

BACKGROUND: Airway management during resuscitation is pivotal for treating hypoxia and inducing reoxygenation. This German Resuscitation Registry (GRR) analysis investigated the influence of the type of airway used in patients treated with manual chest compression (mCC) and automated chest compression devices (ACCD) after out-of-hospital cardiac arrest (OHCA). METHODS: Out of 42,977 patients (1 January 2010-30 June 2016) information on outcome, airway management and method of chest compressions were available for 27,544 patients. Hospital admission under cardiopulmonary resuscitation (CPR), hospital admission with return of spontaneous circulation (ROSC), hospital discharge and discharge with cerebral performance categories 1 and 2 (CPC 1,2) were used to compare outcome in patients treated with mCC vs. ACCD, and classified by endotracheal intubation (ETI), initial supraglottic airway device (SAD) changed into ETI, and only SAD use. RESULTS: Outcomes for hospital admission under ongoing CPR, hospital admission with ROSC, hospital discharge and neurologically intact survival (CPC 1,2) for mCC (84.8%) vs. ACCD (15.2%) groups were: 8.4/38.6%, 39.2/27.2%, 10.6/6.8%, 7.9/4.7% (p < 0.001), respectively. Only mCC with SAD/ETI for ever ROSC (OR 1.466, 95% CI: 1.353-1.588, p < 0.001) and mCC group with SAD/ETI for hospital admission with ROSC showed better outcomes (odds ratio [OR] 1.277, 95% confidence interval [CI]: 1.179-1.384, p < 0.001) in comparison to mCC treated with ETI. Compared to mCC/ETI, all other groups were associated with a decrease in neurologically intact survival. CONCLUSION: Better outcomes were found for mCC in comparison to ACCD and ETI showed better outcomes in comparison to SAD only. This observational registry study raised the hypothesis that SAD only should be avoided or SAD should be changed into ETI, independent of whether mCC or ACCD is used.

Histophilus somni Stimulates Expression of Antiviral Proteins and Inhibits BRSV Replication in Bovine Respiratory Epithelial Cells.

Our previous studies showed that bovine respiratory syncytial virus (BRSV) followed by Histophilus somni causes more severe bovine respiratory disease and a more permeable alveolar barrier in vitro than either agent alone. However, microarray analysis revealed the treatment of bovine alveolar type 2 (BAT2) epithelial cells with H. somni concentrated culture supernatant (CCS) stimulated up-regulation of four antiviral protein genes as compared with BRSV infection or dual treatment. This suggested that inhibition of viral infection, rather than synergy, may occur if the bacterial infection occurred before the viral infection. Viperin (or radical S-adenosyl methionine domain containing 2--RSAD2) and ISG15 (IFN-stimulated gene 15--ubiquitin-like modifier) were most up-regulated. CCS dose and time course for up-regulation of viperin protein levels were determined in treated bovine turbinate (BT) upper respiratory cells and BAT2 lower respiratory cells by Western blotting. Treatment of BAT2 cells with H. somni culture supernatant before BRSV infection dramatically reduced viral replication as determined by qRT PCR, supporting the hypothesis that the bacterial infection may inhibit viral infection. Studies of the role of the two known H. somni cytotoxins showed that viperin protein expression was induced by endotoxin (lipooligosaccharide) but not by IbpA, which mediates alveolar permeability and H. somni invasion. A naturally occurring IbpA negative asymptomatic carrier strain of H. somni (129Pt) does not cause BAT2 cell retraction or permeability of alveolar cell monolayers, so lacks virulence in vitro. To investigate initial steps of pathogenesis, we showed that strain 129Pt attached to BT cells and induced a strong viperin response in vitro. Thus colonization of the bovine upper respiratory tract with an asymptomatic carrier strain lacking virulence may decrease viral infection and the subsequent enhancement of bacterial respiratory infection in vivo.

Adverse effects of a 10-day course of ibuprofen in Holstein calves.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are recommended for various conditions in cattle. Ibuprofen is an inexpensive short-acting NSAID and is readily available in liquid formulation for administration to bottle-fed calves. We compared the adverse effects of a 10-day course of ibuprofen and placebo in 16 five- to six-week-old Holstein bull calves that were being treated for experimentally induced bovine respiratory syncytial virus infection. Ibuprofen was administered as a liquid in milk replacer at 30 mg/kg divided three times daily. We found an increased prevalence of abomasal ulceration 5 of 8 in the ibuprofen compared to placebo group 2 of 6 (P = NS). There was one (1 of 8) case of mild interstitial nephritis in the ibuprofen and none (0 of 8) in the placebo group (P = NS). Renal function as measured by serum BUN and creatinine levels was not different between groups; no animal demonstrated an increase in creatinine.

Comparative analysis of zygotic developmental genes in Rhodnius prolixus genome shows conserved features on the tracheal developmental pathway.

Most of the in-depth studies on insect developmental genetic have been carried out in the fruit fly Drosophila melanogaster, an holometabolous insect, so much more still remains to be studied in hemimetabolous insects. Having Rhodnius prolixus sequenced genome available, we search for orthologue genes of zygotic signaling pathways, segmentation, and tracheogenesis in the R. prolixus genome and in three species of Triatoma genus transcriptomes, concluding that there is a high level of gene conservation. We also study the function of two genes required for tracheal system development in D. melanogaster - R. prolixus orthologues: trachealess (Rp-trh) and empty spiracles (Rp-ems). From that we see that Rp-trh is required for early tracheal development since Rp-trh RNAi shows that the primary tracheal branches fail to form. On the other hand, Rp-ems is implied in the proper formation of the posterior tracheal branches, in a similar way to D. melanogaster. These results represent the initial characterization of the genes involved in the tracheal development of an hemimetabolous insect building a bridge between the current genomic era and V. Wigglesworth's classical studies on insects' respiratory system physiology.

[Randomised trial of acupuncture compared with conventional massage and "sham" laser acupuncture for treatment of chronic neck pain - range of motion analysis]. / Randomisierte Studie zur Akupunktur im Vergleich mit konventioneller Massage und Schein-Laserakupunktur in der Behandlung chronischer HWS-Beschwerden - Bewegungsanalyse.

AIM: The aim of this study was to compare the effects of acupuncture on active motion of the cervical spine in patients with chronic neck pain with those of "sham" laser acupuncture and massage. MATERIAL AND METHODS: 177 patients with chronic neck pain were included in this prospective, randomized, placebo-controlled study. The patients were allocated by external randomization to five treatments over three weeks with acupuncture, massage and "sham" laser acupuncture. The range of active motion was measured by means of a 3D ultrasound real time motion analyzer. RESULTS: The analysis of cervical motion in three directions showed the largest increase in range of motion 14 days after acupuncture. Compared to massage, a significant improvement in total range of motion was seen in those patients treated by acupuncture immediately (p = 0,03) and one week (p = 0,03) weeks after therapy. There was no significant difference in those patients treated by sham laser acupuncture. CONCLUSION: The results of the study indicate that acupuncture is superior to conventional massage for improving active range of motion in patients with chronic neck pain. Because of its positive effects, its acceptance among patients and the lack of severe side effects, acupuncture can be recommended for the treatment of chronic neck pain, although there was no significant difference in results between "sham" laser acupuncture and acupuncture.

Vestibular histofluorescence could be due to accumulation of both the antibiotic and its derivative, streptidine, after acute streptomycin treatment in the guinea pig.

Acute treatment with 300 mg/kg of pigmented guinea pigs with streptomycin sulfate induces an elevation of endogenous fluorescence in vestibular ampullary cristae. Fluorescence accumulates in all compartments of the epithelium, i.e., vestibular sensory and supporting cells and nerve fibers of the stroma and it was very intense 1 and 12 hours after its administration. Fluorescence decreased to control levels 24 hours following streptomycin injection. Fluorescence levels were very low either in untreated animals or in animals injected with saline physiological solution. To investigate whether this fluorescence was an intrinsic property of the antibiotic or whether it was due to a derivative of it, or both, an in vitro fluorescence spectrum was performed with 100 microM solutions of streptomycin or streptidine, or both, dissolved in various buffer solutions at 488 nm of excitation. A discrete level of fluorescence was observed in the spectrum regardless of media when separate solutions of both streptomycin or streptidine were studied. Fluorescence notably increased at 522-532 nm when the solutions contained both streptomycin and streptidine together. These results suggest that streptidine putatively derived from streptomycin may contribute to the observed fluorescence accumulation in vestibular preparations after acute treatment. Thus, these metabolic properties of the inner ear which transform streptomycin into streptidine, something never considered earlier, could be claimed as partially responsible for converting a therapeutic agent into a compound which could be as harmful as STP to the inner ear.

Randomised trial of acupuncture compared with conventional massage and "sham" laser acupuncture for treatment of chronic neck pain.

OBJECTIVES: To compare the efficacy of acupuncture and conventional massage for the treatment of chronic neck pain. DESIGN: Prospective, randomised, placebo controlled trial. SETTING: Three outpatient departments in Germany. PARTICIPANTS: 177 patients aged 18-85 years with chronic neck pain. INTERVENTIONS: Patients were randomly allocated to five treatments over three weeks with acupuncture (56), massage (60), or "sham" laser acupuncture (61). PRIMARY OUTCOME MEASURE: maximum pain related to motion (visual analogue scale) irrespective of direction of movement one week after treatment. SECONDARY OUTCOME MEASURES: range of motion (3D ultrasound real time motion analyser), pain related to movement in six directions (visual analogue scale), pressure pain threshold (pressure algometer), changes of spontaneous pain, motion related pain, global complaints (seven point scale), and quality of life (SF-36). Assessments were performed before, during, and one week and three months after treatment. Patients' beliefs in treatment were assessed. RESULTS: One week after five treatments the acupuncture group showed a significantly greater improvement in motion related pain compared with massage (difference 24.22 (95% confidence interval 16.5 to 31.9), P=0.0052) but not compared with sham laser (17.28 (10.0 to 24.6), P=0.327). Differences between acupuncture and massage or sham laser were greater in the subgroup who had had pain for longer than five years (n=75) and in patients with myofascial pain syndrome (n=129). The acupuncture group had the best results in most secondary outcome measures. There were no differences in patients' beliefs in treatment. CONCLUSIONS: Acupuncture is an effective short term treatment for patients with chronic neck pain, but there is only limited evidence for long term effects after five treatments.

Two inhibition targets by [Cr(2gb)(3)](3+) and [Co(2gb)(3)](3+) on redox enzymes of spinach thylakoids.

[Cr(2gb)(3)]Cl(ZnCl(4)), [Cr(2gb)(3)]Cl(3), and [Co(2gb)(3)]Cl(3) were synthesized and characterized. Their chemical structures and the oxidation states of their metal centers remained unchanged in solution. The effects of these compounds, CrCl(3) and [Co(NH(3))(6)]Cl(3), on photosynthesis were investigated. The coordination compounds inhibit ATP synthesis and electron flow (basal, phosphorylating, and uncoupled), behaving as Hill reaction inhibitors. The target for [Cr(2gb)(3)]Cl(ZnCl(4)) is located at the Q(B) level. In contrast, the interaction sites of [Cr(2gb)(3)]Cl(3) and [Co(2gb)(3)]Cl(3) are located in the span from P(680) to Q(A) and at the b(6)f complex. Neither CrCl(3) nor [Co(NH(3))(6)]Cl(3) inhibited photosynthesis. The 100% inhibition on PS II of [Cr(2gb)(3)]Cl(ZnCl(4)) is explained in terms of a synergystic effect between the 2gb-chromium(III) coordination compound and the ZnCl(4)(2)(-) anion.

Detection of human immunodeficiency virus type 1 and type 2 antibodies by a new automated microparticle immunoassay AxSYM HIV-1/HIV-2.

A new automated microparticle enzyme immunoassay (MEIA) for the AxSYM instrument developed recently by Abbott Laboratories was compared with two established assays, i.e. HIV-1/HIV-2 3rd Gen. Plus EIA (Abbott, Delkenheim, FRG) and Wellcozyme HIV 1 + 2 (Murex Diagnostics, Dartford, England) devised for the detection of human immunodeficiency virus type 1 (HIV-1) and HIV-2 antibodies. A total of 7293 serum samples were tested by the AxSYM HIV-1/HIV-2. The test panel included seroconversions (n = 22), samples from HIV-1 and HIV-2 positive individuals, hospitalized patients, blood donors, high risk individuals. To challenge further the specificity of the assays, large numbers of EIA repeatedly reactive but Western blot negative samples, potentially cross-reactive sera, Western blot indeterminate specimens and samples from pregnant women were tested. In four seroconversion panels, HIV-1 infection was detected one bleed earlier with the AxSYM HIV-1/HIV-2 than with the Abbott HIV-1/HIV-2 3rd Gen. Plus EIA. Although the AxSYM HIV-1/HIV-2 was tested with a higher number of challenging sera than the alternative assays, the specificity was very high (99.4%). The highest number of false positive results was obtained with serum samples that were repeatedly reactive in EIAs different from those compared in the present study. The automated AxSYM system permits the testing of a large sample number in a rapid turn-around time and by random access urgent tests can be carried out even when an assay is in progress.

L-651,582 inhibition of intracellular parasitic protozoal growth correlates with host-cell directed effects.

L-651,582 (5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1,2,3-triazole-4- carboxamide), which is active in vivo against coccidiosis caused by the intracellular parasitic protozoan Eimeria tenella, is also effective against this organism in tissue culture in Maden Darby bovine kidney (MDBK) host cells. L-651,582 inhibited 45Ca++ uptake, as well as [3H]hypoxanthine incorporation into soluble nucleotide pools, in MDBK cells at concentrations similar to those required for antiparasitic activity (IC50 = 0.3 microgram/ml). However, the drug did not inhibit the [3H] hypoxanthine incorporation into the nucleotide pools of the intracellular E. tenella under the same conditions. The antiparasitic activity of several L-651,582 analogs paralleled their ability to inhibit [3H]hypoxanthine incorporation in MDBK cells. L-651,582 similarly inhibited the growth of the intracellular parasitic protozoan Toxoplasma gondii in vitro with either HeLa cells or Normal Human Fibroblasts as host cells. The IC50 for inhibition of T. gondii growth in normal human fibroblasts was similar to that for [3H]hypoxanthine incorporation inhibition. In HeLa cells, however, 40-fold higher levels were required for the inhibition of [3H]hypoxanthine incorporation than were required to inhibit parasite growth, showing that antiparasitic activity was not a consequence of the alteration in host nucleotide metabolism. This was also consistent with the observation that the effect of L-651,582 against E. tenella in MDBK cells was not reversed by the addition of nutrients involved in nucleotide biosynthesis. This study suggests that L-651,582 has antiparasitic activity which correlates with host cell effects and which may be a result of blocking Ca++ entry in the host cells, but is not the result of an alteration of host nucleotide biosynthesis.

Coordination compounds derived from the interaction of streptomycin and cobalt, nickel, copper, and calcium salts characterized by 13C NMR and spectroscopic studies. Structure and bonding properties of the streptidine fraction.

The coordination compounds of streptomycin (St), Co2(St)Cl4.13H2O (2), Co2(St)(NO3)4.7H2O (3), Ni2(St)Cl4.14H2O (4), Ni2(St)(NO3)4.14H2O (5), Cu2(St)Cl4.6H2O (6), and Ca(St)Cl2.8H2O (7) have been synthesized by the reaction of streptomycin sulfate (1) with three equivalents of the corresponding inorganic salt. The compounds (2)-(7) were characterized by electronic spectroscopy (in the solid state and in solution) by conductivity measurements and by 13C NMR in solution. The reaction of streptomycin with CuCl2 in water hydrolyzed the molecule giving the copper complex of the streptidine fraction (Std), Cu(Std)Cl.H2O (8). This compound was characterized by the same techniques. Detailed x-ray diffraction and 13C NMR studies of streptidine sulfate (9) were carried out.

Anti-proliferative activity of L-651,582 correlates with calcium-mediated regulation of nucleotide metabolism at phosphoribosyl pyrophosphate synthetase.

L-651,582, 5-amino-[4-(4-chlorobenzoyl)-3,5-dichlorobenzyl]-1, 2,3-triazole-4-carboxamide, is an antiproliferative and antiparasitic agent which inhibits nucleotide metabolism in mammalian cells. The drug equivalently inhibited 3H-hypoxanthine, 14C-adenine, and 14C-formate incorporation into nucleotide pools in Madin-Darby bovine kidney (MDBK) cells, suggesting depletion of the supply of phosphoribosyl pyrophosphate, (PRPP), required for each of these independent pathways. Inhibition of nucleotide metabolism correlated with inhibition of proliferation for three cell types with differing sensitivities toward the drug. L-651,582 inhibited incorporation of 3H-hypoxanthine into nucleotide pools with either glucose, uridine, or ribose as carbon source suggesting a block at PRPP synthetase, rather than a block in a pathway supplying ribose-5-phosphate. PRPP synthetase was not inhibited directly by the compound, indicating regulation of the enzyme in intact cells. Drug treatment did not kill cells but reduced the fraction of cells in S and G2/M while increasing the population in G1. Inhibition of uptake of 45Ca was demonstrated at concentrations identical to those required for inhibition of nucleotide metabolism or proliferation. Inhibition of cellular PRPP biosynthesis rates were also observed using EGTA to lower calcium levels. These data suggest a previously unrecognized link between calcium entry, the regulation of nucleotide biosynthesis at PRPP synthetase, and the rate of proliferation of mammalian cells.

A method for assaying orotate phosphoribosyltransferase and measuring phosphoribosylpyrophosphate.

An orotate phosphoribosyltransferase, OPRTase, assay method which relies upon binding reactant [3H]orotic acid and product [3H]orotidine-5'-monophosphate to polyethyleneimine-impregnated-cellulose resin and collecting on a GFC glass fiber filter is presented. Elution with 2 X 5 ml of 0.1 M sodium chloride in 5 mM ammonium acetate removes all of the orotate and leaves all of the product orotidine monophosphate (OMP) bound so that it may be measured in a scintillation counter. It was found that the addition of 10 microM barbituric acid riboside monophosphate to the reaction mixture prevented the conversion of OMP to UMP and products of UMP. The assay is suitable for measurement of OPRTase activity with purified enzyme or in crude homogenates. A modification of this scheme using commercially available yeast OPRTase and 10 microM of unlabeled OMP provides an assay for phosphoribosylpyrophosphate with a sensitivity such that 10 pmol of PRPP may be measured.

[Effect of homeopathic drugs on the phagocytic activity of human granulocytes. In vitro tests in a controlled single-blind study]. / Die Beeinflussung der Phagozytosefähigkeit von Granulozyten durch homöopathische Arzneipräparate. In-vitro-Tests und kontrollierte Einfachblindstudien.

Four homeopathic drug preparations with extract dilutions between D1 and D30 (expression for the homeopathic grade of potencies) and some additives (minerals and animal toxins) were investigated, using two in vitro and one in vivo phagocytosis models. All preparations enhanced significantly the activity of phagocytosis in all used systems. In controlled prospective single-blind studies with the preparations C and D performed on 12 and 14 verum and 13 male placebo volunteers respectively with i.v. injections within a 5-day treatment the phagocytosis indices were measured during 11 days using the microscopic smear method. In both investigations the maximum of phagocytosis activity was reached between the 4th and 5th day of injection. After the 4th or 5th (next to the last or last) injection, a rapid decrease of activity occurred which reached normal values on the 11th day. Other laboratory parameters investigated were not influenced.

IMP dehydrogenase from the intracellular parasitic protozoan Eimeria tenella and its inhibition by mycophenolic acid.

Mycophenolic acid (MA) was demonstrated to be an effective inhibitor of the growth of the intracellular parasitic protozoan Eimeria tenella in tissue culture and guanine was shown to reverse this inhibition as expected for an inhibitor of IMP dehydrogenase (IMP:NAD+ oxidoreductase, EC 1.1.1.205). A high performance liquid chromatography study of the intracellular nucleotide pools labeled with [3H]hypoxanthine was carried out in host cells lacking hypoxanthine-guanine phosphoribosyltransferase, and the depletion of guanine nucleotides demonstrated that the intracellular parasite enzyme was being inhibited by the drug. Kinetic studies carried out on the enzyme derived from E. tenella oocysts demonstrated substrate inhibition by NAD and mycophenolic acid inhibition similar to that found for mammalian enzymes, but different from that for bacterial enzymes. The inhibition by mycophenolic acid was not time-dependent and was immediately reversed upon dilution. As found previously for other IMP dehydrogenases, an Ordered Bi-Bi mechanism prevails with IMP on first followed by NAD, NADH off first, and then XMP. The kinetic patterns are consistent with substrate inhibition at high concentrations of NAD due to the formation of an E X XMP X NAD complex. Uncompetitive inhibition by MA versus IMP, NAD, and K+ was found and this was interpreted as evidence for the formation of an E X XMP X MA complex. A speculative mechanism for the inhibition of the enzyme is offered which is consistent with the fact that E X XMP X MA readily forms, whereas E X IMP X MA does not.

Aggregation of human neutrophils by heparin.

Commercial heparin preparations induce the aggregation of human polymorphonuclear neutrophils (PMNs). The minimum aggregating concentration (MAC) of different heparin lots was measured under standardized conditions. It was found to vary between 0.3 and 15 units/ml. Gel filtration of heparin showed that the aggregating activity eluted in a sharp peak with a pattern different from heparin. When heparin was bound to antithrombin III-Sepharose, the aggregating activity eluted totally in the high-affinity fraction. When PMNs are partially aggregated with heparin, further aggregation by N-formyl-methionyl-leucyl-phenylalanine (FMLP) or phorbol myristate-acetate (PMA) is not affected. Pretreatment of PMNs with a low-aggregating heparin fraction, inhibits further aggregation by a high-aggregating heparin fraction. These results suggest that the PMNs have binding sites for heparin different from those for PMA and FMLP. Binding of heparin is not enough for inducing aggregation. Certain structural requirements of the heparin molecule are probably essential for aggregation.