Effects of heparin on platelet aggregation and release and thromboxane A2 production.

Heparin, when added to citrated platelet-rich plasma (PRP), caused potentiation of platelet aggregation and the release reaction induced by the aggregating agents adenosine diphosphate (ADP), arachidonic acid, collagen, and epinephrine. At low concentrations (4.7 x 10(-5) M) arachidonic acid failed to cause aggregation of platelets in citrated PRP. However, in the presence of heparin, the same concentration of arachidonic acid caused aggregation. Examination of PRP for the presence of thromboxane A2 (TxA2) by use of a bioassay revealed that heparin also stimulated release of TxA2. This finding indicated that platelets released more TxA2 when they were challenged by low concentrations of arachidonic acid in the presence of heparin than in its absence. Platelets were labeled with 3H-arachidonic acid and 14C-serotonin, and attempts were made to determine whether heparin stimulated the platelet release reaction first with subsequent increased production of TxA2, or alternatively, whether heparin stimulated TxA2 production first with subsequent enhancement of the release reaction. In view of the demonstrated simultaneous release of 14C-serotonin and 3H-arachidonic acid metabolites, it appeared that either release of 14C and 3H occurs concurrently or, even if one of these events is dependent on the other, both events take place in rapid succession. Timed sequential studies revealed that in the presence of arachidonic acid, the addition of heparin hastened the apparently simultaneous release of both 14C and 3H.     

High rates of clustering of strains causing tuberculosis in Harare, Zimbabwe: a molecular epidemiological study

We examined the pattern of tuberculosis (TB) transmission (i.e., reactivation versus recent transmission) and the impact of human immunodeficiency virus (HIV) infection in Harare, Zimbabwe. Consecutive adult smear-positive pulmonary TB patients presenting to an urban hospital in Harare were enrolled. A detailed epidemiological questionnaire was completed, and tests for HIV type 1 and CD4 cell counts were performed for each patient. Molecular fingerprinting of the genomic DNA recovered from cultures of sputum was performed by two molecular typing methods: spacer oligonucleotide typing (spoligotyping) and analysis of variable number of tandem DNA repeats (VNTRs). A cluster was defined as isolates from two or more patients that shared the same spoligotype pattern or the same VNTR pattern, or both. DNA suitable for typing was recovered from 224 patients. The prevalence of HIV infection was 79%. Of 187 patient isolates (78.6%) typed by both spoligotyping and analysis of VNTRs, 147 were identified as part of a cluster by both methods. By spoligotyping alone, 84.1% of patient isolates were grouped into 20 clusters. The cluster size was generally <8 patient isolates, although three large clusters comprised 68, 25, and 23 patient isolates. A total of 89.4% of the patient isolates grouped into 12 clusters defined by analysis of VNTRs, with 2 large clusters consisting of 127 and 13 patient isolates, respectively. Thirty-six percent of patient isolates with a shared spoligotype and 17% with a shared VNTR pattern were geographically linked within Harare, but they were not linked on the basis of the patient's home district. In a multivariate analysis, there were no independent predictors of clustering, including HIV infection status. Comparison with the International Spoligotype database (Pasteur Institute, Pointe a Pitre, Guadeloupe) demonstrated that our three largest spoligotype clusters are well recognized and ubiquitous in Africa. In this epidemiologically well characterized urban population with a high prevalence of HIV infection, we identified a very high level of strain clustering, indicating substantial ongoing recent TB transmission. Geographic linkage could be detected in a proportion of these clusters. A small group of actively circulating strains accounted for most of the cases of TB transmission.     

Postsynaptic fall in intracellular pH and increase in surface pH caused by efflux of formate and acetate anions through GABA-gated channels in crayfish muscle fibres.

H(+)-selective microelectrodes and a two- or three-microelectrode voltage clamp were used to examine the influence of weak-acid, carboxylate anions on the actions of GABA on postsynaptic intracellular pH, surface pH and on membrane potential in fibres of the crayfish leg opener muscle. Substitution of 30 mM Cl- by formate or acetate promoted a GABA-induced decrease in intracellular pH, which was coupled to an increase in surface pH and to a depolarization. Such effects were not seen in the presence of an equivalent amount of lactate, methanesulphonate or glucuronate. Both the GABA-induced depolarization and the fall in internal pH promoted by formate and acetate were blocked by picrotoxin, and the fall in pH was reversibly inhibited by a K(+)-induced depolarization. The rate of the fall in intracellular pH produced by GABA (0.2 mM) was about 0.02 pH units/min in the presence of formate and 0.03 pH units/min in the presence of acetate. Under steady-state conditions, both 30 mM formate and acetate (but not lactate) induced a positive shift in the reversal potential of GABA-activated current, which was accounted for by a relative permeability vs Cl- of formate and acetate of 0.5 and 0.15, respectively. The conductance sequence of the anions was identical to the permeability sequence, i.e. Cl- greater than formate greater than acetate greater than lactate approximately equal to 0. This sequence is strictly correlated to the Stokes diameter of the anions. The relative permeabilities of the anions indicate that the effective diameter of the GABA-gated channel is about 0.5 nm. The fact that the GABA-induced acidosis was slower in the presence of formate than in the presence of acetate suggests that, in the former case, the rate-limiting step in the fall in internal pH is the entry of non-dissociated formic acid. All the above results are consistent with a scheme where GABA induces a channel-mediated efflux of permeant weak-acid anions, which gives rise to an inward (depolarizing) current and to an intracellular acidosis. A comparison of the permeability properties of crayfish and vertebrate GABA-gated channels suggests that effects similar to those seen in this work are likely to occur in mammalian and other vertebrate neurons in the presence of permeant weak-acid anions.     

Lactoferrin in aspirates of odontogenic cyst fluid.

The possibility that the presence of lactoferrin in aspirates of odontogenic cyst fluid might be a useful preoperative diagnostic marker for odontogenic keratocyst was investigated. Using qualitative and quantitative immunodiffusion methods fluid from 29 of 29 dental (radicular) cysts, 12 of 14 dentigerous cysts and 27 of 31 keratocysts were found to contain lactoferrin. Although some of the highest concentrations of lactoferrin were detected in fluids from keratocysts, there was no significant difference between lactoferrin concentrations among the three groups. Neutrophil elastase was detected in 20 of 24 samples tested, 22 of which also contained lactoferrin. Immunocytochemical localisation of both lactoferrin and elastase was confined to neutrophils infiltrating cyst walls. These results suggest that lactoferrin in fluid from odontogenic cysts is derived from infiltrating neutrophils and that its presence in aspirated fluids is not a useful diagnostic marker for odontogenic keratocyst.     

烧伤的流行病学、人口统计学及结局特点

老年人的群体在不断扩大，而他们烧伤的发病率和死亡率都要比年轻人高。最近的一项病例回顾研究分析了一家烧伤中心7年来住院患者的资料，结果显示，1557位住院患者中有221位（11％）年龄在59岁以上（含59岁）。其中有97位（44％）是女性，反映出老年患者中女性比例较高。     

In vitro histamine release induced by radiocontrast media and various chemical analogs in reactor and control subjects

The factors governing in vitro basophil histamine release induced by radiocontrast material (RCM) were evaluated by the use of two RCMs (diatrizoate and metrizamide) and three structural analogs of RCM (benzoic acid, diaminobenzoic acid, and 3-acetamidobenzoic acid) in basophil histamine-release studies. Diatrizoate was chosen because it is a commonly used RCM and is routinely administered as a hypertonic drug (958 mOsm/kg or greater). Metrizamide is a newly synthesized RCM and is routinely administered in isotonic form (approximately 280 mOsm/kg). The analogs were used in hope of identifying any structure-function relationship that might exist between RCMs and the activation of a proposed basophil membrane receptor responsible for the induction of de granulation. In addition, results in a control group of normal subjects were compared with those in a group of patients who had experienced anaphylactoid reactions to intravenous RCM. Basophils from both groups were incubated with diatrizoate and metrizamide to assess their relative sensitivity to these drugs. Both metrizamide and diatrizoate induced in vitro histamine release. Therefore hypertonicity is not an absolute requirement for RCM-induced in vitro basophil de granulation. Although there was a trend far reagents without a prosthetic group at position 5 on the benzene ring (especially 3-acetamidobenzoic acid) to induce more release of histamine than reagents with such a prosthetic group, differences between these reagents did not reach statistical significance. Therefore no clearcut structure-function relationship could be demonstrated. “Reactor” subjects released significantly-larger percents of their intracellular histamine than did “nonreactors” (p < 0.05). Degranulated cells retained their ability to exclude trypan blue, an observation suggesting that RCM-induced histamine release does not involve cell death.     

Antibody L26 recognizes an intracellular epitope on the B-cell-associated CD20 antigen.

Monoclonal antibody L26 is a highly selective marker of B cells and B-cell neoplasms in paraffin-embedded tissues, but it suffers from the drawback that the target molecule has not been identified. In this paper we provide evidence by two independent techniques that antibody L26 recognizes an intracellular epitope on the CD20 antigen (a pan B-cell marker). When this antigen was redistributed on the surface of unfixed viable B cells by incubation with monoclonal anti-CD20 followed by anti-mouse Ig, the diffuse cytoplasmic staining of L26 was abolished and replaced by coincident dotlike labeling for antibody L26 and the CD20 antigen. None of the other antibodies tested (covering 10 different B-cell-associated antigens) had this effect on the L26 staining pattern. Furthermore, COS-1 cells transfected with cDNA encoding the CD20 molecule gave positive staining with antibody L26 and with two other CD20 reagents, but not with antibodies to other pan B-cell markers (eg, CD19 and CD22).     

Proliferation in non-Hodgkin''s lymphoma: a comparison of Ki-67 staining on fine needle aspiration and cryostat sections.

Assessment of the growth fraction of non-Hodgkin's lymphomas may provide useful additional prognostic information to that obtained with conventional histological criteria. The monoclonal antibody Ki-67 has been reported to provide such information immunocytochemically in tissue biopsy specimens from lymphoma as well as other tumours. This study was undertaken to assess whether this approach could be extended to fine needle aspiration (FNA) biopsy specimens which are becoming increasingly important in the diagnosis of lymphoma. In 21 cases of non-Hodgkin's lymphoma the rate of tumour proliferation estimated by Ki-67 immunostaining of FNA material, obtained from surgically removed specimens, was compared with that obtained on tissue biopsy. The correlation between both preparations was excellent, indicating that FNA biopsy material is suitable for the immunocytochemical assessment of the growth fraction of non-Hodgkin's lymphoma.     

Frequency and properties of naturally occurring adherent piliated strains of Haemophilus influenzae type b.

We found that 41 of 75 (55%) children with Haemophilus influenzae type b disease (70 cases of meningitis, 2 of cellulitis, 2 of septic arthritis, and 1 of epiglottitis) and 2 of 120 (1.7%) children with upper respiratory infection were colonized with H. influenzae type b in the nasopharynx (NP). Of these 43 NP strains from children with systemic H. influenzae type b disease, 7 (16%) adhered to human buccal epithelial cells. The strains isolated from the systemic site of all children, including children from whose NP adherent bacteria were isolated, did not adhere to buccal epithelial cells in vitro. Each adherent NP strain had biotype (I), serotype (b), and antibiotic susceptibility (sensitive) similar to that of the corresponding nonadherent systemic isolate. With one exception, all NP-systemic pairs had similar major outer membrane proteins. Six of the seven NP strains had a protein band in the whole cell lysate preparation with a molecular weight between 22,000 and 23,000, which could not be seen in the nonadherent cerebrospinal fluid strains. Electron micrographs of all adherent strains showed that more than 95% of the organisms examined were highly piliated, whereas the nonadherent strains were not piliated. All piliated strains agglutinated human erythrocytes. Adherence to buccal epithelial cells and agglutination of erythrocytes could not be blocked by mannose or alpha-methyl-D-mannoside. We speculate that piliation is not important for NP colonization by H. influenzae type b and that the loss of pili may be required for host invasion.     

Human Leucocyte Differentiation Antigen nomenclature: update on CD nomenclature. Report of IUIS/WHO Subcommittee

The 7th International Workshop on Human Leucocyte Differentiation Antigens (HLDA7) studied a number of newly characterised molecules relevant to human leucocyte differentiation and function. The HLDA organisation, which devised and continues to maintain the CD nomenclature, is responsible, under the auspices of IUIS and WHO, for the nomenclature of all leucocyte differentiation markers. The 7th Workshop redefined a number of (principally carbohydrate) molecules, and assigned CD names to approximately 80 new molecules. This update lists, in tabular form, the redefined and newly assigned names, together with antibodies, which have been confirmed under Workshop conditions as specific for the new and redefined molecules. The major features of the cellular expression patterns are summarised, and a LocusLink accession number provided to enable the reader to access more detailed information through http://www.ncbi.nlm.nih.gov/LocusLink.