Search PubMed⌕ Search

Biomedical subjects

J Kun

Publications and source records attributed to J Kun.

16 recordsLinked to original sources
PubMed2001

A method for measuring passive elastance during proportional assist ventilation.

There are currently no reliable, noninvasive ways to monitor respiratory elastance (E) during assisted ventilation. We describe a method that is suited for proportional assist ventilation (PAV). In this mode, the end of the ventilator's inflation phase occurs during the declining phase of inspiratory effort (Pmus). If the opening of the exhalation valve is delayed, airway pressure (Paw) should initially rise as Pmus continues its decline. When Pmus declines to zero, a Paw plateau should appear. Paw at this point should reflect passive recoil at the prevailing volume. A cohort of 74 ventilator-dependent patients, ventilated in the PAV mode, were studied. Brief end-inspiratory occlusions were applied at random intervals. The magnitude of early change in Paw during the occlusion was inversely related to level of assist (r = 0.7, p < 0.00001). At high assist (> 75%), Paw was nearly flat or declined slightly, indicating minimal residual Pmus at the onset of occlusion. At lower assist levels, Paw increased exponentially in most patients with an average time constant of 0.21 +/- 0.06 s. Extraneous events that may corrupt the measurement (e.g., behavioral responses) were extremely rare (< 0.5%) in the first 0.25 s. From these findings, we concluded that Paw measured 0.25 s from occlusion onset (P0.25) includes little inspiratory Pmus and is free of extraneous events. E, estimated from P0.25 during PAV (EPAV), agreed well (r = 0.92) with passive E measured during controlled ventilation (ECMV); the average difference (EPAV - ECMV) was (+/- SD) -0.3 +/- 4.9 cm H2O x L(-1), corresponding to 0.9 +/- 16.4% of average E. We conclude that Paw measured at 0.25 s from the onset of end-inspiratory occlusion in the PAV mode provides a reliable estimate of passive elastic recoil.

Adult↗
PubMed2001

Variants of Plasmodium falciparum erythrocyte membrane protein 1 expressed by different placental parasites are closely related and adhere to chondroitin sulfate A.

Plasmodium falciparum-infected erythrocytes adhere to syncytiotrophoblast cells lining the placenta via glycosaminoglycans, such as chondroitin sulfate A (CSA) and hyaluronic acid. Adherence of infected erythrocytes to host receptors is mediated by P. falciparum erythrocyte membrane protein-1 (PfEMP-1). A single PfEMP-1 domain (duffy binding-like [DBL]-3, of the gamma sequence class) from laboratory-adapted strains is thought to be responsible for binding to CSA. In this study, DBL-gamma domains expressed by placental P. falciparum isolates were shown to have an affinity to CSA. All parasite populations accumulating in infected placentas express only 1 variant of PfEMP-1, each of which contains a DBL-gamma domain with CSA binding capacities. Furthermore, sequence analysis data provide evidence for antigenic conservation among the DBL-gamma sequences expressed by different placental parasites. This study offers a close reflection of the process of parasite adhesion in the placenta and is crucial to the understanding of the pathogenesis of malaria during pregnancy.

Amino Acid Sequence↗
PubMed2001

A method for noninvasive determination of inspiratory resistance during proportional assist ventilation.

Currently available noninvasive methods for measuring inspiratory resistance (RI) are difficult to implement or interpret during assisted mechanical ventilation on account of the confounding effect of respiratory efforts (Pmus). We propose a simple method consisting of brief reductions in airway pressure (Paw) in the early part of the inflation phase (pulse). Paw, flow (V), and volume (V) are measured at the beginning of the pulse (T (0)), at the trough of the pulse (TI) and at a point 0.1 s before T(0) (T(-1)). Equations of motion of the form [Pmus + Paw = V. K(1) + V (2). K(2) +V. E] are generated for the data at the three time points (E = elastance, K(1) and K(2) are Rohrer's constants). These three equations can be solved for K(1) and K(2) if it is arranged that the pulse has appropriate configuration and timing, and if it is assumed that DeltaPmus/Deltat is constant over the brief pulse period. The method was tested in 67 patients ventilated with proportional assist ventilation (PAV). The results were compared with those obtained using the interrupter technique during a period of controlled mechanical ventilation (CMV). RI, expressed at a standard flow of 1 L. s(-)(1), was slightly higher during PAV (16.4 +/- 4.9 versus 15.5 +/- 4.5 cm H(2)O. L(-1). s, p < 0.001). The average difference was 0.9 +/- 2.0 cm H(2)O. L(-1). s, corresponding to 5.4 +/- 12.6% of the average of RCMV and RPAV. The correlation coefficient was 0.92 (p = 8E-28) with a slope (1.01) and intercept (0.8) not significantly different from 1.0 and 0, respectively. We conclude that brief negative pulses applied early during the inflation phase can be used to provide reliable estimates of inspiratory resistance during PAV.

Airway Resistance↗
PubMed1999

Stimulation of CD95 (Fas) blocks T lymphocyte calcium channels through sphingomyelinase and sphingolipids.

Calcium influx through store-operated calcium release-activated calcium channels (CRAC) is required for T cell activation, cytokine synthesis, and proliferation. The CD95 (Apo-1/Fas) receptor plays a role in self-tolerance and tumor immune escape, and it mediates apoptosis in activated T cells. In this paper we show that CD95-stimulation blocks CRAC and Ca(2+) influx in lymphocytes through the activation of acidic sphingomyelinase (ASM) and ceramide release. The block of Ca(2+) entry is lacking in CD95-defective lpr lymphocytes as well as in ASM-defective cells and can be restored by retransfection of ASM. C2 ceramide, C6 ceramide, and sphingosine block CRAC reversibly, whereas the inactive dihydroceramide has no effect. CD95-stimulation or the addition of ceramide prevents store-operated Ca(2+) influx, activation of the transcriptional regulator NFAT, and IL-2 synthesis. The block of CRAC by sphingomyelinase metabolites adds a function to the repertoire of the CD95 receptor inhibiting T cell activation signals.

Animals↗
PubMed1997

Evidence for a novel function of the CD40 ligand as a signalling molecule in T-lymphocytes.

The interaction of the CD40 receptor with its ligand has been shown to be crucial for the activation of B-lymphocytes. Here, we provide evidence that the pg39 molecule/CD40 ligand (gp39/CD40L) also functions as a stimulatory molecule for T-lymphocytes. Activation of T-lymphocytes via gp39/CD40L induced a strong activation of Jun-N-terminal kinase (JNK) and p38-K. Activation of these kinases correlates with a stimulation of Rac1 and inhibition of Rac1 prevents gp39/CD40L triggered JNK/p38-K activation. Further, cellular stimulation via the CD40 ligand results in tyrosine phosphorylation of cellular proteins and the activation of p56(lck). Inhibition of src-like kinases inhibits Rac1 as well as JNK/p38-K stimulation suggesting a signalling cascade from the gp39/CD40L via p56(lck) and Rac1 to JNK/p38-K.

Animals↗
PubMed1997

Expression of the Sox11 gene in mouse embryos suggests roles in neuronal maturation and epithelio-mesenchymal induction.

Sry, the mammalian Y-linked testis determining gene, is a member of a family of genes known as Sox genes, which encode transcription factors related by a DNA-binding motif termed the HMG box. Sox genes are known to have diverse roles in vertebrate differentiation and development. We report here the cloning and characterisation of one of these genes, Sox11, in mice. In addition to an N-terminal HMG box domain, the deduced SOX11 protein contains a number of highly conserved C-terminal motifs, which may function in transcriptional regulation. Expression of Sox11 in mouse embryos was prominent in the periventricular cells of the central nervous system, suggesting a role in neuronal maturation. Expression was also observed in a wide range of tissues involved in epithelial-mesenchymal interactions, suggesting an additional role in tissue modelling during development.

Amino Acid Sequence↗
PubMed1995

The Sry-related gene Sox9 is expressed during chondrogenesis in mouse embryos.

Mutations in the human SRY-related gene, SOX9, located on chromosome 17, have recently been associated with the sex reversal and skeletal dysmorphology syndrome, campomelic dysplasia. In order to clarify the role of this gene in skeletal development, we have studied the expression of mouse Sox9 during embryogenesis. Sox9 is expressed predominantly in mesenchymal condensations throughout the embryo before and during the deposition of cartilage, consistent with a primary role in skeletal formation. Interspecific backcross mapping has localized mouse Sox9 to distal chromosome 11. The expression pattern and chromosomal location of Sox9 suggest that it may be the gene defective in the mouse skeletal mutant Tail-short, a potential animal model for campomelic dysplasia.

Amino Acid Sequence↗
PubMed1995

Glutamic acid decarboxylase-67 (GAD67): expression relative to GAD65 in human islets and mapping of autoantibody epitopes.

Glutamic acid decarboxylase (GAD), a target of both autoantibodies and autoreactive T-cells in insulin-dependent diabetes (IDD), exists as two homologous forms, GAD65 and GAD67. GAD65 is preferentially expressed in human islets and recognized by autoantibodies in IDD, but which form primarily elicits GAD autoimmunity is unknown. GAD67 gene expression in human islets has been demonstrated only by the polymerase chain reaction. We, therefore, quantitatively compared the expression of each GAD gene in human islets and mapped the binding of autoantibodies to recombinant human GAD67 by enzyme-linked immunosorbent assay. In ribonuclease protection assays, both forms of GAD messenger RNA (mRNA) were detected in human islets, although GAD65 mRNA was 200 times more abundant than GAD67 mRNA. Immunoblotting of islets with GAD form-specific antisera revealed GAD65, but not GAD67. By in situ hybridization and immunohistochemistry, GAD65 mRNA and protein were localized to islets, predominantly, but not entirely, to beta-cells; GAD67 mRNA and protein were undetectable. Thus, although GAD67 protein expression was undetectable in human islets, the GAD67 gene is transcribed, albeit weakly. Antibodies that recognized multiple epitopes in recombinant GAD67 were found in 20% of sera from ICA positive "at risk" first degree relatives of IDD subjects and recent-onset IDD subjects. The majority of GAD67 epitopes were mapped within the mid- and C-terminal thirds of the protein, a region that is highly conserved in GAD65. Although GAD67 may share cross-reactive epitopes with GAD65, these findings do not exclude the possibility that autoimmunity to GAD arises as a consequence of the aberrant up-regulation of GAD67 in human islets.

Autoantibodies↗
PubMed1993

In vitro phagocytosis inhibition assay for the screening of potential candidate antigens for sub-unit vaccines against the asexual blood stage of Plasmodium falciparum.

We have previously established a direct correlation between immune protection against the asexual blood stage Plasmodium falciparum infection and the presence of opsonizing antibodies promoting phagocytosis of parasitized red blood cells. In the present communication we describe an in vitro assay for measuring phagocytosis inhibition (PIA) specific for P. falciparum-infected erythrocytes. The phagocytosis inhibition assay is a simple procedure for screening potential candidates for sub-unit vaccines against P. falciparum based on the correlation between opsonizing antibodies and immunoprotection. The assay was used to analyse 18 recombinant molecules, corresponding to 11 distinct antigens of P. falciparum. Pre-incubation and selective antibody depletion experiments demonstrate the antigen-antibody specificity of the PIA. The presence of epitopes participating as targets of opsonic antibodies were demonstrated in six distinct polypeptide antigens.

Animals↗
PubMed1993

cDNA cloning and expression of Rhodnius prolixus vitellogenin.

It was shown that Rhodnius prolixus vitellogenin (Vg) is synthesized as precursors of 205 and 190 kDa. Each Vg subunit is antigenically related to a domain in the precursor molecules. Since Vg has been previously detected in R. prolixus male adults, protein synthesis by fat bodies from 5th instar male nymphs was investigated and no Vg synthesis could be detected. Also, a 6.1 Kb RNA is present in female adults but not in 5th instar male nymphs. Therefore, cDNAs from female adult and 5th instar male fat bodies were used for differential screening of a female fat body cDNA library leading to the isolation of several female specific clones. All the clones hybridizing to the female specific 6.1 Kb RNA species were identical. We also describe the construction of new expression vectors, pGex-A and pGex-B, derived from the previously described plasmid pGex-1N. The new vectors, together with pGex-3X, comprise a set of expression plasmids with cloning sites in all three possible reading frames that give a fusion polypeptide with the glutathione S-transferase. This carrier protein can be cleaved by digestion with factor Xa in all three plasmids; one of the Vg cDNA clones was subcloned in pGex-A. Antibodies affinity purified from the fusion protein Vg/glutathione S-transferase recognized both large Vg subunits, suggesting an antigenic relationship between them. Furthermore, the small Vg subunits were not recognized, indicating that they may be localized at the N-terminal region of Vg precursors.

Amino Acid Sequence↗
PubMed1993

Localization and quantitation of expression of two glutamate decarboxylase genes in pancreatic beta-cells and other peripheral tissues of mouse and rat.

Glutamic acid decarboxylase (GAD) catalyzes synthesis of the inhibitory neurotransmitter gamma-amino butyric acid. Two homologous forms of GAD encoded by separate genes have been cloned from rat brain, with predicted protein sizes of 67 and 65 kilodaltons. GAD is present outside the brain, and pancreatic islet GAD is believed to be a target of autoimmunity in insulin-dependent diabetes mellitus. However, peripheral expression of the two GAD genes is incompletely characterized. We, therefore, investigated GAD expression in peripheral tissues, including pancreas, of mouse and rat. cDNAs encoding GAD 67 and GAD 65 were cloned from mouse brain and shown to be 95% homologous with the rat sequences. RNase protection assay using specific cRNA probes demonstrated expression of both GAD forms in freshly harvested pancreas and testis. Levels of both GAD mRNAs were greater in rat than mouse pancreas. GAD 67 mRNA was more abundant than GAD 65, and both were localized to islet beta-cells by in situ hybridization. In testis, both GAD mRNAs were localized to spermatocytes. Additionally, GAD 67, but not GAD 65, mRNA was detected in mouse and rat spleen and mouse liver. Thus, both GAD genes are expressed in peripheral tissues, with GAD 67 mRNA being more abundant under physiological conditions. The expression of both GAD 67 and GAD 65 genes specifically in islet beta-cells indicates that both GAD forms are candidate autoantigens in rodent models of insulin-dependent diabetes mellitus.

Animals↗
PubMed1989

A versatile phage lambda expression vector system for cloning in Escherichia coli.

By integrating fragments from the expression plasmids pJK2 and pJK4 into a derivative of the bacteriophage lambda, we constructed the phage expression vectors lambda JK2 and lambda JK4, which allow efficient cloning of genomic or cDNA either into the 5' end or the 3' end of the lacZ gene of Escherichia coli. Expression of barrier-free DNA in phase may lead to fusion proteins consisting of active beta-galactosidase (beta Gal) plus an additional polypeptide encoded by the inserted DNA. Analysis of distinct recombinant clones is quick and easy, due to the reversible integration of the plasmid into the genome. As an example, we constructed an expression library of genomic Plasmodium falciparum DNA in lambda JK2. We polymerised (amplified) and expressed a synthetic DNA fragment, which codes for a potential antigenic determinant of the 11-1 gene of Plasmodium falciparum as a fusion to the N terminus of active beta Gal. We demonstrate that such chimeric molecules can be affinity-purified and that polypeptides can be separated from the beta Gal part by cleavage with the protease factor Xa.

Amino Acid Sequence↗
PubMedDate not supplied

Cloning and expression of genomic DNA sequences coding for putative erythrocyte membrane-associated antigens of Plasmodium falciparum.

Genomic DNA fragments of Plasmodium falciparum generated by mung bean nuclease digestion were cloned in the lambda expression vector lambda JK2. The resulting library was screened with a rabbit antiserum raised against purified membranes of P. falciparum-infected erythrocytes and with a serum pool from immune humans from an endemic area of Liberia. Positive clones were rescreened with a series of human and monkey sera. Twelve selected clones were analysed in detail. Four of them corresponded to already described membrane-associated P. falciparum antigens. The other positive clones contained inserts which, according to the nucleotide sequence, Southern blot analysis and immunological characteristics, correspond to so far unknown antigens.

Amino Acid Sequence↗
PubMedDate not supplied

[Usefulness of the fluorescein dilaurate test in patients after acute pancreatitis].

30 patients, 1 to 6 years acute pancreatitis, complaining on epigastric pains, weight loss, flatulence and diarrhea were examined. Temmler Werke Pancreolauryl-Test was performed following manufacturer's instructions. In 14 patients (46.7%), the result of the test revealed pancreatic exocrine failure. Those results were verified and confirmed in 80% by secretin-ceorulein test. Non-invasive, technically uncomplicated and reliable pancreolauryl-test seems to be useful for screen estimation of pancreatic exocrine function impairment in patients after the acute pancreatitis.

Acute Disease↗