A brief introduction of the Chinese Marrow Donor Program.

The Data Bank of Chinese Hematopoietic Stem Cell Donors (Chinese Marrow Donor Program, CMDP) is operated under the guidance of Red Cross Society of China. It is in charge of the administration of propagation, organisation, mobilisation of volunteers, and the standardisation of human leukocyte antigen (HLA) typing, preliminary search, clinical transplantation, and all related activities in unrelated stem cell transplantation in China. The Advisor Committee--composed by experts in the fields of haematopoietic stem cell transplantation (HSCT), HLA typing, legislation, information technology and ethics--guides the development of the CMDP. The budget of CMDP mainly comes from funds of National Charity Lottery and other charities. Up to the end of 2008, 31 branch registries and HLA-typing laboratories, five high-resolution laboratories, and one quality-control laboratory were established and authorized. There are more than 950,000 donors in our data pool. The CMDP has established clinical relationship with over 100 hospitals. More than 1100 CMDP donors have donated peripheral blood stem cells to patients successfully. The CMDP has also signed cooperation agreement with five of the seven major cord blood banks in mainland China. As a result, there are over 30,000 units of cord blood available for searching in our system. This will improve the matching and using rate of cord blood. The emergence of the CMDP has filled a void in mainland China's contribution to clinical HSCT and donor search worldwide. It has performed preliminary searches for overseas patients including Taiwan, Hong Kong, Macao, United States, Britain, Canada, France, Italy, Germany, Japan, Korea and Singapore, and over 50 donations have been completed.

Molecular evidence that aphid-transmitted Alpinia mosaic virus is a tentative member of the genus Macluravirus.

Alpinia mosaic virus (AlpMV), once assigned to the genus Potyvirus, infects primarily plants of the ginger family. To seek molecular evidence for correct classification of this virus, a cDNA clone corresponding to the 3' portion of the AlpMV genome was obtained by reverse transcriptase-PCR and TA cloning. The authenticity of the cDNA clone was confirmed by expression of the coat protein (CP) in E. coli followed by immunoblot analysis. Sequence analysis indicated that, in contrast to its low identity with all the other genera of the family Potyviridae, the deduced amino acid sequence of AlpMV CP was 42.9 - 61.9% identical to members of the genus Macluravirus. Phylogenetic analysis also demonstrated that the AlpMV CP clustered with those of Cardamom mosaic virus and Chinese yam necrotic mosaic virus. These results indicate that AlpMV should be classified as a tentative species within the genus Macluravirus rather than Potyvirus as proposed previously.

Prostaglandin E(2) enhances chemical and mechanical sensitivities of pulmonary C fibers in the rat.

It has been recently reported that pulmonary reflex responses to injection or inhalation challenge of capsaicin are enhanced by exogenous Prostaglandin E(2) (PGE(2)). The present study was carried out to determine whether PGE(2) enhances the stimulatory effects of chemical stimulants and lung inflation on vagal pulmonary C fibers, and if so, whether the excitabilities of other types of lung afferents are also augmented by PGE(2). In anesthetized, open-chest rats, administration of PGE(2) (1.5 microgram/kg/min for 2 min) did not significantly change the baseline activity of vagal pulmonary C fibers, but it markedly enhanced the stimulatory effects of both low (0.25 microgram/kg) and high doses (0.5 microgram/kg) of capsaicin on these fibers. Similarly, potentiating effects of PGE(2) were found on the pulmonary C-fiber responses to injections of lactic acid and adenosine, although considerable variability existed in the degrees of potentiation between the different stimulants. Furthermore, PGE(2) infusion also significantly enhanced the C-fiber response to constant-pressure lung inflation (tracheal pressure [Pt] = 30 cm H(2)O). In contrast, PGE(2) did not alter the responses of either slowly adapting pulmonary receptors or rapidly adapting pulmonary receptors to lung inflation. In summary, these results show that the sensitivity of pulmonary C-fiber afferents to both mechanical and chemical stimuli is enhanced by PGE(2), suggesting that endogenous release of this autocoid may play a part in the airway irritation and dyspneic sensation associated with airway inflammation.

Activation of pulmonary C fibres by adenosine in anaesthetized rats: role of adenosine A1 receptors.

1. Intravenous administration of adenosine (Ado) to patients can cause dyspnoea, chest discomfort and bronchoconstriction. To assess the role of vagal pulmonary C fibres in evoking these adverse reactions, the effect of Ado on single pulmonary C fibres was studied in anaesthetized and artificially ventilated rats. 2. Right-atrial injection of Ado (320 microg kg-1) activated 68 % (73/107) of pulmonary C fibres; the total number of action potentials during a period of 15 s increased from a baseline of 0.2 +/- 0.1 impulses to a peak of 16.4 +/- 2.6 impulses (P < 0.01, n = 107) after Ado. Inosine, the metabolite of Ado, did not activate any of eleven C fibres tested in six rats. Furthermore, C fibres were activated only by right-atrial and not by left-ventricular injection of the same dose of Ado. 3. Unlike the immediate and transient stimulation of C fibres by capsaicin, the C fibre stimulation by Ado had a latency of 6.5 +/- 0.3 s (range, 3-18 s) and lasted longer. 4. The stimulation of C fibres by Ado was significantly attenuated by pretreatment with aminophylline, a non-selective Ado receptor antagonist, was completely prevented by 1,3-dipropyl-8-cyclopentylxanthine, an Ado A1 receptor antagonist, but was unaffected by 3,7-dimethy-1-propargylxanthine, an A2 receptor antagonist. None of these Ado receptor antagonists prevented capsaicin-induced C fibre stimulation. 5. In conclusion, Ado stimulates pulmonary C fibre terminals through an activation of A1 receptors. The stimulation of pulmonary C fibres may play an important role in Ado-induced adverse respiratory effects.

Role of pulmonary C fibers in adenosine-induced respiratory inhibition in anesthetized rats.

The clinical use of adenosine is commonly associated with pulmonary side effects, namely dyspnea, that suggest the possible involvement of bronchopulmonary sensory afferents. Our objective in this study was to characterize the effects of adenosine on breathing and to determine whether the vagal pulmonary afferents play a role in mediating these effects. We measured respiratory and cardiovascular changes in anesthetized, spontaneously breathing rats after bolus injections of adenosine at therapeutic doses. Right atrial injection of adenosine (0.04-0.6 mg/kg) elicits, in a dose-dependent manner, a pulmonary chemoreflex-like response consisting of a delayed apnea, bradycardia, and hypotension. In contrast, the classic capsaicin-elicited pulmonary chemoreflex occurs immediately after injection. Perineural capsaicin treatment of the cervical vagi blocked the adenosine-induced respiratory inhibition. Left ventricular administration of adenosine failed to elicit an apneic response. Pretreatment with the adenosine A1-receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine attenuated the adenosine-induced apnea. These results indicate that adenosine elicits a respiratory inhibition via stimulation of pulmonary C fibers and that activation of the A1-receptor is probably involved. It is unclear, however, what accounts for the exceedingly long latency in this response.

Stimulation of pulmonary C fibres by lactic acid in rats: contributions of H+ and lactate ions.

1. The contributions of H+ and lactate ions to the stimulation of single pulmonary C fibres by lactic acid were examined in anaesthetized and artificially ventilated rats. 2. Lactic acid injected into the right atrium caused a transient decrease in arterial blood pH (pHa) and a short but intense burst of afferent activities in pulmonary C fibres, whereas sodium lactate had no effect. The fibre activity usually reached a peak within 1-1.5 s, with an onset latency of < 1 s, and returned to the baseline in 5 s. 3. The injection of hydrochloric acid at the same pH as that of lactic acid did not significantly decrease pHa, nor did it stimulate any C fibres studied. 4. Formic acid has a pKa value (the negative logarithm of the dissociation constant) almost identical to that of lactic acid; thus, its injection decreased pHa to the same degree as did the injection of lactic acid. However, the response of C fibres to lactic acid was 134% stronger than that to formic acid. 5. We conclude that H+ is primarily responsible for the activation of pulmonary C fibres by lactic acid, probably through a direct effect of H+ on these afferent endings. The lactate ion, by itself, does not activate C fibres, but it seems to potentiate the stimulatory effect of H+ on these afferents.

Cigarette smoke-induced bronchoconstriction: causative agents and role of thromboxane receptors.

Inhalation of cigarette smoke induces a biphasic bronchoconstriction in guinea pigs: the first phase is induced by a combination of cholinergic reflex and tachykinins, whereas the second phase involves cyclooxygenase metabolites (J.-L. Hong, I. W. Rodger, and L.-Y. Lee. J. Appl. Physiol. 78: 2260-2266, 1995). This study was carried out to further determine the causative agents in the smoke and the types of prostanoid receptors and endogenous prostanoids mediating the bronchoconstriction. Inhalation of 10 ml of high-nicotine cigarette smoke consistently elicited the biphasic bronchoconstriction in anesthetized and artificially ventilated guinea pigs. Pretreatment with hexamethonium (10 mg/kg iv) significantly reduced the first-phase bronchoconstriction but did not have any measurable effect on the second-phase response. In sharp contrast, gas-phase smoke did not elicit any bronchoconstrictive effect. Furthermore, when the animals were challenged with low-nicotine cigarette smoke, only a single second-phase response was evoked, accompanied by increases in thromboxane (Tx) B2 (a stable metabolite of TxA2), prostaglandin (PG) D2, PGF2 alpha in the bronchoalveolar lavage fluid. The bronchoconstrictive response induced by low-nicotine smoke was completely prevented by pretreatment with SQ-29548 (0.3 mg/kg iv), a TxA2-receptor antagonist. These results indicate that 1) nicotine is the primary causative agent responsible for the first-phase bronchoconstriction and 2) nonnicotine smoke particulates evoke the release of TxA2, PGD2, and PGF2 alpha, which act on TxA2 receptors on airway smooth muscles and induce the second-phase response to cigarette smoke.

Cigarette smoke-induced bronchoconstriction: cholinergic mechanisms, tachykinins, and cyclooxygenase products.

The mechanisms underlying cigarette smoke-induced bronchoconstriction were studied by using selective blockade of muscarinic acetylcholine receptors, neurokinin receptors and production of eicosanoids of the cyclooxygenase pathway in anesthetized guinea pigs. Inhalation of three breaths of cigarette smoke (University of Kentucky research series 2R1; 2.45 mg of nicotine and 35.3 mg of tar per cigarette) reproducibly induced an immediate bronchoconstriction; total pulmonary resistance increased from 0.24 +/- 0.02 to 1.44 +/- 0.21 cmH2O.ml-1.s (P < 0.01) and dynamic lung compliance decreased from 0.53 +/- 0.03 to 0.39 +/- 0.06 ml/cmH2O (P < 0.05) in 10-15 breaths after the smoke inhalation. Atropine pretreatment (50 micrograms/kg i.v.) prevented the immediate decrease in dynamic lung compliance and reduced the immediate increase in total pulmonary resistance by approximately 55%. The atropine-resistant bronchoconstriction occurring immediately after smoke inhalation was completely blocked by a pretreatment with a combination of CP-99994 (0.3 mg/kg i.v.) and SR-489668 (0.3 mg/kg i.v.), the antagonists of neurokinin-1 and neurokinin-2 receptors, respectively. However, a delayed and sustained bronchoconstriction still persisted and reached a plateau in 45-55 breaths after smoke inhalation challenge. This delayed response was completely prevented by pretreatment with indomethacin (5 mg/kg i.v.). We conclude that the smoke-induced bronchoconstriction in guinea pigs consists of an early phase induced by both a cholinergic reflex and tachykinin release, probably evoked by the activation of bronchopulmonary C fibers, and a late phase caused by the action of arachidonic acid metabolite(s) of the cyclooxygenase pathway.

Cigarette smoke-induced bronchoconstriction and release of tachykinins in guinea pig lungs.

Two series of experiments were carried out to determine whether the release of tachykinins is involved in the bronchoconstriction induced by inhalation of cigarette smoke in guinea pigs. In the first series, cigarette smoke consistently induced bronchoconstriction (delta RL = +203% and delta Cdyn = -46%) in anesthetized guinea pigs, and the response was only partially blocked by bilateral cervical vagotomy. However, the smoke-induced bronchial constriction was completely abolished in animals receiving a systemic capsaicin pretreatment to destroy the tachykinin-containing C-fiber afferents. In the second series, the bronchoconstrictive effect of cigarette smoke was increased by approx. three times in isolated perfused guinea pig lungs when phosphoramidon (3 x 10(-6) M) was added to the perfusate to prevent the degradation of tachykinins after their release. Moreover, the enhanced bronchomotor response to smoke was accompanied by an overflow of neurokinin A-like immunoreactivity (LI) and calcitonin gene-related peptide -LI in the pulmonary effluent. These studies showed that cigarette smoke triggers the release of tachykinins in the lungs, which plays an important role in the smoke-induced bronchoconstrictive effect in guinea pigs.

Modification of glutathione S-transferase 3-3 mutants with 2-(S-glutathionyl)-3,5,6-trichloro-1,4-benzoquinone. Identification of the C-terminal tryptic fragment as part of the H-site and evidence that 2-(S-glutathionyl)-3,5,6-trichloro-1,4-benzoquinone is not specific for cysteine labelling.

A triple mutant of rat liver glutathione S-transferase 3-3 that has all three cysteine residues replaced with serine (CallS) and a quadruple mutant with a Tyr-115 to phenylalanine substitution on CallS (CallSY115F) were reacted with 2-(S-glutathionyl)-3,5,6-trichloro-1,4-benzoquinone (GS-1,4-TCBQ). The modified proteins were analysed on a triple-quadrupole mass spectrometer equipped with an electrospray ionization source. At an enzyme: GS-1,4-TCBQ ratio of 1:10, the enzymes were modified at multiple sites. Covalent attachment of a single inhibitor on to the protein was achieved by lowering the enzyme: GS-1,4-TCBQ ratio to 1:1. Results from m.s. analyses suggest that the inhibitor on the CallSY115F mutant exists as a glutathionyl dichlorobenzoquinone derivative. The modifiers of the CallS mutants are glutathionyl monochlorobenzoquinone derivatives. Therefore, GS-1,4-TCBQ reacts at a single site on CallSY115F, but probably cross-links two regions on wild-type and CallS mutant. To confirm our observation, CallS was modified with 1-chloro2,4-dinitrobenzene, which specifically labels Tyr-115, before reacting with GS-1,4-TCBQ. The inhibitor formed a glutathionyl dichlorobenzoquinone adduct on the dinitrophenyl-CallS mutant. In addition, the benzoquinone derivative on the protein can be partially removed by 1-chloro-2,4-dinitrobenzene. Peptide mapping and sequencing analysis of the GS-1,4-TCBQ-modified CallS mutant revealed that the C-terminal 16-amino-acid fragment is labelled. Molecular modelling suggests the C(5) and C(6) on the benzoquinone ring of the inhibitor interact with the oxygen atoms of Tyr-115 and Ser-209 respectively.

Reversible modification of rat liver glutathione S-transferase 3-3 with 1-chloro-2,4-dinitrobenzene: specific labelling of Tyr-115.

Rat liver glutathione S-transferase 3-3 (GST, EC 2.5.1.18), a triple mutant with all three cysteine residues replaced with serine (CallS) and a quadruple mutant with a Tyr-115 to phenylalanine substitution on CallS (CallSY115F) were overexpressed in Escherichia coli under the control of a phoA promoter. Using this system, we obtained over 35 mg of fully active pure protein/litre of cell medium. GST 3-3 and CallS mutant were modified with 1-chloro-2,4-dinitrobenzene (CDNB), a model substrate for the enzyme, in the absence of GSH. Dinitrophenol, but not S-methylglutathione, inhibits this process. The dinitrophenyl groups are readily removed from the enzyme with GSH, but much more slowly with dithiothreitol. Results from peptide mapping and amino acid sequence analyses indicate that CDNB modifies the cysteine residues and Tyr-115 on wild-type GST 3-3, but only Tyr-115 on CallS. In addition, CDNB cannot modify the CallSY115F mutant. We propose that Tyr-115 is located at or near the H-site of GST 3-3.

Site-directed mutagenesis and chemical modification of cysteine residues of rat glutathione S-transferase 3-3.

Rat liver glutathione S-transferase (GST) 3-3 is composed of two identical subunits, each containing three cysteine residues, Cys-86, Cys-114 and Cys-173. We have shown previously that Cys-86 is not involved in the enzymic activity of GST 3-3 [Hsieh, Huang, Chen, Lai & Tam (1991) Biochem, J. 278, 293-297]. At 50 degrees C, iodoacetamide can inactivate the enzyme by modifying Cys-86 and Cys-114. Cys-114 can be protected against iodoacetamide inhibition by S-(dinitrophenyl)glutathione. Site-directed mutagenesis was used to construct mutants in which serine replaced one (C114S and C173S) or all three (CallS) cysteine residues. These mutants were over-expressed in Spodoptera frugiperda cells in a baculovirus system and were found to be fully active. Replacing Cys-86 or Cys-114 with alanine (C86A and C114A) does not diminish the activity of the protein. The results suggest that cysteines are not involved in the enzymic mechanism, and Cys-114 is possibly located at the active site of GST 3-3.

[Effects of tetrandrine on cytoplasmic free calcium in rat neutrophils].

Neutrophils (NP) cytoplasmic free Ca2+ concentrations ([Ca2+]i) under several conditions were measured with Quin 2 in rats. The process of loading Quin 2-AM into the cells followed by its hydrolysis was assured by monitoring the shift of fluorescent emission spectrum of Quin 2-AM to that of Quin 2. The resting [Ca2+]i of NP in Ca2(+)-containing and Ca2(+)-free solutions were 186 +/- 45 and 46 +/- 16 nmol/L, respectively, indicating extracellular calcium concentration ([Ca2+]0) plays an important role on [Ca2+]i. Among agents tested, prostaglandin E2 (PGE2) 100 nmol/L did not change [Ca2+]i significantly. Calcimycin 25 mumol/L, leukotriene B4(LTB4) 60 nmol/L and platelet activating factor (PAF) 10 nmol/L increased [Ca2+]i of NP in Ca2(+)-containing solution from 185 +/- 54, 175 +/- 36 and 188 +/- 54 to 814 +/- 67, 577 +/- 229 and 540 +/- 174 nmol/L, respectively. But, compound 48/80 3.2 micrograms/ml, LTB4 300 nmol/L, PAF 10 nmol/L and PAF 5 nmol/L plus LTB4 150 nmol/L did not change significantly [Ca2+]i of NP in Ca2(+)-free solution, indicating that these agonists can not release intracellularly stored Ca2+ or no stored Ca2+ in NP is available. The rises of [Ca2+]i produced by calcimycin, PAF and LTB4 were markedly inhibited by tetrandrine (Tet) 65 mumol/L. These results, as a whole, show that Tet inhibits the rises of [Ca2+]i of NP induced by calcimycin, PAF and LTB4 via decreasing Ca2+ influx. The Ca2+ antagonism in this case may be related to its anti-allergic actions.

[Variability of Apo B-containing lipoprotein in the plasma of experimental atherosclerotic rabbits].

The effects of dietary cholesterol on plasma Apo B-containing lipoprotein was studied in 18 rabbits. When atherosclerosis was induced by dietary cholesterol, the cholesterol and peroxide lipids were increased in the intima of aortae (P less than 0.05). The increased plasma Apo B was positively correlated with hypercholesterolemia (r = 0.879, P less than 0.005). In the pre-stain lipoprotein electrophoresis and immunoelectrophoresis with Apo B antiserum, electrophoretic mobility of Apo B-containing lipoprotein was slower than those of the control rabbits and this band was wide and stained deeper. In Sephadex G-200 exclusion chromatography there was evidence showing that the reduction in electrophoretic mobility of Apo B-containing lipoprotein is due to the increase particle size.

Variation in ten lysosomal hydrolase enzyme activities in inbred mouse strains.

Activities of 10 lysosomal hydrolase enzymes (beta-hexosaminidase, beta-galactosidase, alpha-galactosidase, alpha-mannosidase, beta-mannosidase, alpha-L-fucosidase, beta-glucuronidase, alpha-glucosidase, alpha-N-acetylgalactosaminidase, and acid phosphatase) were determined in eight organs (brain, liver, kidney, spleen, heart, skeletal muscle, lung, and testis) in males and females of six inbred mouse strains (C57BL/6J, C3H/HeJ, DBA/2J, BALB/cJ, P/J, and 129/J). Examples of enzyme-specific variation, organ-specific variation, and enzyme- and organ-specific variation were found. New enzyme-specific variants with the features of systemic regulators for alpha-L-fucosidase and beta-mannosidase were found. Known variants were detected. Organ-specific variants had some of the properties expected for a new class of genes affecting multiple enzymes: organ-specific regulators.

Variation in alpha-L-fucosidase properties among 28 inbred mouse strains: six strains have high enzyme activity and heat-stabile enzyme with a variant pH-activity curve; twenty-two strains have low activity and heat-labile enzyme.

Alpha-L-fucosidase in tissues of 28 inbred mouse strains varied with respect to three properties: high or low heat stability, a pH-activity curve with high or low relative activity at pH 2.8, and high or low activity. Alpha-L-fucosidase from six strains (A/J, BDP/J, LP/J, P/J, SEA/GNJ, and 129/J) had high heat stability, high pH 2.8 relative activity, and high activity, whereas the other 22 strains all had low heat stability, low pH 2.8 relative activity, and low activity. The heat-stability difference was seen in all organs tested (brain, liver, kidney, spleen, heart, skeletal muscle, lung, and testis) for two heat-stabile strains (P/J and 129/J) and four heat-labile strains (C57BL/6J, C3H/HeJ, DBA/2J, and BALB/cJ) studied in detail. The findings suggested that two structural variants of alpha-L-fucosidase, probably genetically determined, exist in these 28 inbred mouse strains, although the presence of linkage disequilibrium between alleles of tightly linked structural and regulatory genes could not be excluded.

Human lymphedema fluid lipoproteins: particle size, cholesterol and apolipoprotein distributions, and electron microscopic structure.

The concentration of cholesterol, apolipoproteins A-I, B, and E has been determined in lymphedema fluid from nine patients with chronic primary lymphedema. The concentrations were: 38.14 +/- 21.06 mg/dl for cholesterol, 15.6 +/- 6.17 mg/dl for apolipoprotein A-I, 7.5 +/- 2.8 mg/dl for apolipoprotein B, and 1.87 +/- 0.50 mg/dl for apolipoprotein E. These values represent 23%, 12%, 6%, and 38% of plasma concentrations, respectively. The ratio of esterified to unesterified cholesterol in lymphedema fluid was 1.46 +/- 0.45. Lipoproteins of lymphedema fluid were fractionated according to particle size by gradient gel electrophoresis and by exclusion chromatography. Gradient gel electrophoresis showed that a majority of high density lipoproteins (HDL) of lymphedema fluid were larger than ferritin (mol wt 440,000) and smaller than low density lipoproteins (LDL); several discrete subpopulations could be seen with the large HDL region. Fractionation by exclusion chromatography showed that more than 25% of apolipoprotein A-I and all of apolipoprotein E in lymphedema fluid was associated with particles larger than plasma HDL2. Apolipoprotein A-I also eluted in fractions that contained particles the size of or smaller than albumin. Isolation of lipoproteins by sequential ultracentrifugation showed that less than 25% of lymphedema fluid cholesterol was associated with apolipoprotein B. The majority of apolipoprotein A-containing lipoproteins of lymphedema fluid were less dense than those in plasma. Ultracentrifugally separated fractions of lipoproteins were examined by electron microscopy. The fraction d less than 1.019 g/ml contained little material, while fraction d 1.019-1.063 g/ml contained two types of particles: round particles 17-26 nm in diameter and square-packing particles 13-17 nm on a side. Fractions d 1.063-1.085 g/ml had extensive arrays of square-packing particles 13-14 nm in size. Fractions d 1.085-1.11 g/ml and fractions d 1.11-1.21 g/ml contained round HDL, 12-13 nm diameter and 10 nm diameter, respectively. Discoidal particles were observed infrequently.

Comparison of apoprotein B of low density lipoproteins of human interstitial fluid and plasma.

Virtually all apoprotein B (apoB)-containing lipoproteins of the peripheral interstitial fluid of subjects with primary lymphoedema float in the ultracentrifugal field in the density interval 1.019-1.063 g/ml; in this respect they are similar to plasma low-density lipoproteins (LDL). 2. Virtually all apo-B-containing lipoproteins of interstitial fluid migrate in the electrophoretic field with pre-beta mobility; in this respect they are similar to plasma very-low-density lipoproteins. 3. The apoB of lipoproteins of interstitial fluid does not differ in terms of Mr from apoB-100 of human plasma [Kane, Hardman & Paulus (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 2465-2469] as determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 4. Both apoB of interstitial fluid and plasma are heterogenous in terms of their charge as determined by isoelectric focusing of their complexes with the nonionic detergent Nonidet P40. ApoB of plasma LDL focuses between pH5.9 and 6.65, and that of interstitial fluid LDL between pH 5.9 and 6.1. Thus the overall charge of apoB of interstitial fluid is more negative than that of its plasma LDL counterpart.