First national conference on organ specific alloantigens.
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Animals↗
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Isolated total lung perfusion as a means to deliver organ-specific chemotherapy: long-term studies in animals.
The objectives of this study were to develop a surgical procedure that would allow for bilateral isolated lung perfusion in vivo as a means of delivering organ-specific chemotherapy and to evaluate the influence of the procedure on certain pulmonary physiologic parameters. The sterile surgical procedure that was carried out in dogs involved the setting up of two separate perfusion circuits. Once standard systemic cardiopulmonary bypass was established, a second circuit was devised to perfuse the lungs by placing an inflow cannula into the main pulmonary artery and collecting venous effluent in the left atrium. Cross-contamination between perfusion circuits was determined in acute studies with labeled plasma protein or red blood cells and was found to be in an acceptable range if the aorta was cross-clamped and the heart arrested. Only about 0.4 ml/min of pulmonary perfusate leaked into the systemic circulation, indicating that systemic toxicity should not be a major concern when chemotherapy agents are added to the pulmonary perfusate. Chronic studies demonstrated that hemodynamic parameters, lung water, pulmonary endothelial serotonin extraction, and histologic findings all showed minimal changes after 50 minutes of isolated lung perfusion. Five days after perfusion, lung dynamic compliance and peak serotonin extraction showed significant decreases. However, all of the measured parameters had returned toward baseline levels by the end of the 8-week postoperative study period. The procedure offers significant advantages over the previously described single lung perfusion and may provide a method of delivering immediate high-concentration adjuvant chemotherapy to coincide with resection of primary or metastatic lung tumors.
Animals↗
Organ-specific expression of highly divergent thionin variants that are distinct from the seed-specific crambin in the crucifer Crambe abyssinica.
Most thionins of higher plants are toxic to various bacteria, fungi, and animal and plant cells. The only known exception is the seed-specific thionin, crambin, of the crucifer Crambe abyssinica. Crambin has no net charge, is very hydrophobic and exhibits no toxicity. In the present work, the organization of the crambin precursor polypeptide was deduced from cD-NA sequences. The precursor shows a domain structure similar to that of the preproprotein of other thionins, which contains a signal peptide, a thionin domain and a C-terminal amino acid extension. Unlike the thionin precursors studied thus far, both the thionin domain and the C-terminal amino acid extension of the crambin precursor have no net charge and are hydrophobic, thus facilitating their interaction, by analogy to that proposed for the corresponding domains of other thionin precursors that have positive and negative charges. The existence of a large number of novel and highly variable thionin variants in Crambe abyssinica has been deduced from cDNA sequences that were amplified by the polymerase chain reaction (PCR) from RNA of seeds, leaves and cotyledons. While the deduced amino acid sequences of the thionin domains of most of these thionin precursor molecules are highly divergent, the two other domains are conserved. Most of the predicted thionin variants are positively charged. The presence of positively charged residues in the thionin domains consistently correlates with the presence of a negatively charged residue in the C-terminal amino acid extension of the various thionin precursors. The different thionin variants are encoded by distinct sets of genes and are expressed in an organ-specific manner.
Amino Acid Sequence↗
Organ specific neoantigens reactive in the leukocyte adherence inhibition assay: affinity purification of human colon carcinoma antigen and its cross-reactive protein using monoclonal antibodies.
A monoclonal antibody (mAb) was prepared against a semipurified preparation of an organ specific neoantigen (OSN) reactive in the leukocyte adherence inhibition (LAI) assay. The mAb (LC20.1) induces a positive LAI response when incubated with leukocytes of normal individuals in the presence of OSN derived from either human colon or lung carcinoma cell lines. Absorption of crude OSN preparations from these cell lines on immobilized LC20.1 mAb eliminates all the LAI reactive material suggesting that the mAb recognizes a common determinant on OSN from both colon and lung carcinomas. The LC20 mAb was used to affinity purify the colon cancer OSN as well as a cross-reactive normal protein from the urine of colon cancer patients and healthy donors, respectively. The colon cancer OSN and normal cross-reactive protein display an apparent molecular weight of 29,000, have a similar linear tryptic peptide map, and are indistinguishable by isoelectric focusing analysis. Regardless of the molecular similarity, only the colon cancer OSN preparation could induce a positive LAI when incubated with leukocytes of colon cancer patients. Seven additional anti-colon cancer OSN mAbs were prepared against purified material. These mAbs can be divided into three groups, each of which recognizes a distinct antigenic determinant that is shared by the colon cancer neoantigen and its cross-reactive normal protein.
Animals↗
The tomato RBCS3A promoter requires integration into the chromatin for correct organ-specific regulation.
In tomato, the RBCS1, RBCS2 and RBCS3A genes, encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, are expressed in leaves and light-grown seedlings, but only RBCS1 and RBCS2 are expressed in developing tomato fruits. The activities of the three promoters have been compared in transgenic plants and after transient transformation. Fruit-specific repression of the RBCS3A promoter was observed in transgenic plants, but not after ballistic transient transformation, indicating that chromatin integration is necessary for its correct organ-specific regulation. In addition, matrix attachment regions have been identified in the RBCS1, RBCS2 and RBCS3A promoters. This is the second case in plants of absence of correct regulation of a plasmid-borne plant promoter and correlating potential nuclear matrix attachment of the gene.
Antigens, Nuclear↗
Target specific organization and neuron types of the dog pelvic ganglia: a retrograde-tracing and immunohistochemical study.
The major pelvic ganglion in both the rat and guinea pig has been extensively studied because of its anatomical simplicity. To clarify the target specific neural pathway in the diffusely distributed pelvic ganglia of larger animals, the pelvic plexus of the female dog was investigated by retrograde tracing and immunohistochemistry. The whole mount staining of the pelvic plexus with acetylcholinesterase histochemistry revealed 70-100 ganglia of varying sizes. Neurons retrogradely labeled from the rectum were mainly found in ganglia located in the dorso-caudal part of the plexus. The majority of these were non-catecholaminergic, immunoreactive for either calbindin (Calb) or neuropeptide Y (NPY), and characteristically associated with baskets of enkephalin (ENK)-immunoreactive varicose fibers. Neurons projecting to the utero-vaginal walls were distributed in ganglia located in the ventro-caudal part of the plexus. These mainly consisted of two major neuron groups: catecholaminergic Calb-immunoreactive neurons, and non-catecholaminergic neurons containing nitric oxide synthase (NOS) and/or vasoactive intestinal peptide (VIP), which were preferentially associated with a network of ENK-immunoreactive varicose fibers. Neurons retrogradely labeled from the urinary bladder mainly occurred in ganglia located around the junction between the ureter and the bladder. These consisted of catecholaminergic Calb neurons and noncatecholaminergic neurons containing Calb or NOS. Only a few ENK-immunoreactive fibers were found within the clusters of catecholaminergic neurons. These results indicate that organ specific neurons are located in separate ganglia and have both a distinctive composition of neuron types as well as different innervation by preganglionic fibers.
Animals↗
Impact of parietal cell autoantibodies and non-organ-specific autoantibodies on the treatment outcome of patients with hepatitis C virus infection: a pilot study.
AIM: Various side effects have been reported in patients infected with hepatitis C virus (HCV) who were treated with interferon-alpha (IFN-alpha), including the appearance or exacerbation of underlying autoimmune diseases and the development of a variety of organ and non-organ specific autoantibodies (NOSA). However, very few studies in adults have been strictly designed to address: whether the prevalence and the titre of organ and NOSA in serial samples of HCV-treated patients were affected by IFN-alpha, and the impact of these autoantibodies on the treatment outcome of HCV patients. METHODS: We investigated whether parietal cell autoantibodies (PCA) and/or NOSA were related with treatment-outcome in 57 HCV-treated patients (19 sustained-responders, 16 relapsers, 22 non-responders). Serum samples from patients were studied blindly at three time-points (entry, end of treatment and end of followup). For the detection of autoantibodies we used indirect immunofluorescence, commercial and in-house ELISAs. RESULTS: Sustained biochemical response was associated with ANA-negativity at the entry or end of follow up. Sustained virological response was associated with the absence of PCA at the entry. Combined virological and biochemical sustained response (CVBSR) was associated with the absence of antinuclear antibodies (ANA) at the end of follow up and PCA-negativity at the entry. Sustained virological and CVBSR were associated with a reduction of ANA and SMA titers during therapy. CONCLUSION: Although PCA and/or NOSA seropositivity should not affect the decision to treat HCV patients, the presence of some of them such as ANA, PCA and SMA before treatment or their increase during therapy with IFN-alpha may predict a worse response, indicating the need for a closer monitoring during treatment of HCV patients positive for these autoantibodies.
Adult↗
Identification and characterization of novel rat and human gonad-specific organic anion transporters.
We have isolated three novel organic anion transporter cDNAs designated rat GST-1 (gonad-specific transporter), rat GST-2, and human GST, expressed at high levels in the testis. Rat GST-1, GST-2, and human GST consist of 748, 702, and 719 amino acids, respectively, and all molecules possess the 12 predicted transmembrane domains, which is a common structure of organic anion transporters. Northern blot analyses and in situ hybridization revealed that both of the rat molecules are highly expressed in the testis, especially in Sertoli cells, spermatogonia, and Leydig cells. Weak signals are also detected in the epididymis and ovary in adult rat. The exclusive expression of human GST mRNA in the testis was confirmed by RT-PCR. The pharmacological experiments of Xenopus laevis oocytes injected with the respective rat GST-1- and GST-2-cRNAs revealed that both rat GST-1 and GST-2 transport taurocholic acid, dehydroepiandrosterone sulfate, and T4 with Michaelis-Menten kinetics (taurocholic acid, Km = 8.9 and 2.5 microm, dehydroepiandrosterone sulfate, Km = 25.5 and 21.microm, and T4, Km = 6.4 and 5.8 for rat GST-1 and GST-2, respectively). T3 was also transported by rat GST-1 and GST-2. These data suggest that rat GST-1 and GST-2 might be one of the molecular entities responsible for transporting dehydroepiandrosterone sulfate and thyroid hormones involved in the regulation of sex steroid transportation and spermatogenesis in the gonad.
Amino Acid Sequence↗
Organ-specific change in Dolichos biflorus lectin binding by myocardial endothelial cells during in vitro cultivation.
Endothelial cells of the NMRI mouse strain express a cell surface glycoprotein recognized by the lectin Dolichos biflorus agglutinin (DBA). This study documents a marked organ-specific increase in DBA-specific lectin binding of myocardium-derived endothelial cells (MEC) of the NMRI/GSF mouse during in vitro cultivation. An up to 20-fold increase in DBA binding sites is observed in long-term culture, an increase not found in other NMRI-derived endothelial cell lines (e.g., brain, aorta). The increase appears restricted to DBA in that binding with other lectins (PNA, WGA) was unaltered. NMRI MEC cultures maintain typical endothelial cell attributes such as cobblestone morphology on confluence, expression of endothelial cell-specific surface markers, and production of angiotensin-converting enzyme. Cultures routinely become aneuploid within 4 passages, several passages before upregulation of the DBA binding site(s). Myocardial endothelial cells sorted to obtain DBAhi and DBAlo cell populations generally maintained their sorted phenotype for 3 to 4 passages. Limiting dilution cloning resulted in clones varying in DBA expression. Clones for DBAhi expression maintained their DBA affinity for at least 10 passages (> 30 doublings), whereas DBAlo clones gave rise to varying numbers of DBAhi cells within 2 to 4 passages. We hypothesize that the change in DBA affinity accompanies in vitro aging, that the change is independent of alterations in karyotype, and that the increase in DBA affinity may reflect a change in one or more other endothelial cell properties. Additional studies will be necessary to determine whether the in vitro changes are correlated with specific functional alterations and whether they accurately reflect progressive changes of MEC in vivo.
Animals↗
Organ-specific analysis of the anaerobic primary metabolism in rice and wheat seedlings. I: Dark ethanol production is dominated by the shoots.
During anaerobiosis in darkness the main route for ATP production in plants is through glycolysis in combination with fermentation. We compared the organ-specific anaerobic fermentation of flooding-tolerant rice (Oryza sativa) and sensitive wheat (Triticum aestivum) seedlings. A sensitive laser-based photoacoustic trace gas detection system was used to monitor emission of ethanol and acetaldehyde by roots and shoots of intact seedlings. Dark-incubated rice seedlings released 3 times more acetaldehyde and 14 times more ethanol than wheat seedlings during anaerobiosis. Ninety percent of acetaldehyde originated from shoots of both species. In comparison to wheat shoots, the high ethanol production of rice shoots correlated with larger amounts of soluble carbohydrates, and higher activities of fermentative enzymes. After 24 h of anaerobiosis in darkness rice shoots still contained 30% of aerated ATP level, which enabled seedlings to survive this period. In contrast, ATP content declined almost to zero in wheat shoots and roots, which were irreversibly damaged after a 24-h anaerobic period. When plants were anaerobically and dark incubated for 4 h and subsequently transferred back to aeration, shoots showed a transient peak of acetaldehyde release indicating prompt re-oxidation of ethanol. Post-anoxic acetaldehyde production was lower in rice seedlings than in wheat. This observation accounts for a more effective acetaldehyde detoxification system in rice. Compared to wheat the greater tolerance of rice seedlings to transient anaerobic periods is explained by a faster fermentation rate of their shoots allowing a sufficient ATP production and an efficient suppression of toxic acetaldehyde formation in the early re-aeration period.
Acetaldehyde↗
[Non-organ specific anticytoplasmic autoantibodies. Immunoserologic detection and diagnostic relevance].
The detection of circulating serum antibodies that react with various nuclear or cytoplasmic antigens of cell substrates has become an established investigation in the diagnosis of autoimmune diseases. Antinuclear antibodies as well as some non-organ-specific anticytoplasmic autoantibodies might be of diagnostic significance. Some examples are anti-mitochondrial antibodies in autoimmune liver diseases, anti-cytoskeletal antibodies in chronic active hepatitis, polymyositis and infectious diseases and microsomal antibodies in juvenile cryptogenic liver cirrhosis. New techniques and methods such as indirect immunofluorescence on tissue culture substrates, enzymelinked immunoassays and immunoblotting tests now form the basis of further studies. These tests allow a more sensitive and specific assay for lysosomal and Golgi apparatus antigens. It may be possible to more precisely associate particular anti-cytoplasmic antibodies that react with autoimmune diseases.
Antibody Specificity↗
Cloning, expression, and ontogeny of mouse organic anion-transporting polypeptide-5, a kidney-specific organic anion transporter.
The full-length coding sequence of mouse organic anion-transporting polypeptide (designated mouse Oatp-5) has been cloned from mouse kidney cDNA library. Analysis of the 5'-untranslated region (5'-UTR) of Oatp-5 cDNA through capsite cloning reveals two possible transcription start sites that are 4-bp apart. The 3'-untranslated region (3'-UTR) of Oatp-5 cDNA contains an early polyadenylation signal, indicating the possibility that mRNAs with different 3'-UTR lengths may coexist. Deduced amino acid sequence of mouse Oatp-5 protein contains 670 amino acids and has 10 putative transmembrane domains, multiple potential glycosylation and phosphorylation sites. Tissue-specific expression studies indicate that mouse Oatp-5 is expressed only in kidney. Studies on the developmental expression reveal that there is no significant expression of Oatp-5 mRNA in mouse kidney for at least 3 weeks after birth, and adult levels of Oatp-5 mRNA expression are attained more than 6 weeks after birth. Phylogenetic analysis reveals that mouse Oatp-5 is an ortholog of rat Oatp-5.
Aging↗
Non-organ-specific and tumour-specific antibodies in children with Wilms' tumour.
Sera from groups of children with Wilms' tumour, with non-renal solid tumours, and from control children were studied by immunofluorescence and by absorption techniques. A high proportion of sera from both tumour groups contained non-organ-specific auto-antibodies. Three of the 45 sera in the Wilms' group and one of the 27 sera in the "other" solid tumour group contained antibody reacting with normal kidney cells. Only one of 45 sera in the Wilms' tumour group contained tumour-specific antibody.
Adolescent↗
Organ-specific genotoxic effects of chemicals: the use of alkaline elution to detect DNA damage in various organs of in vivo-exposed animals.
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Animals↗
Organ-specific autoimmunity in patients with premature ovarian failure.
The diagnosis of premature ovarian failure (POF) was made in 45 women (aged 14 to 39 yr) with high concentrations of FSH and LH. Sera from these patients and a control group of 28 women were used to detect organ-specific autoantibodies. Eighteen patients with POF (40%) were positive for at least one autoantibody. In the control group only one woman (3.6%) showed autoimmunity. Antithyroid was the most representative autoimmunity (20%) in the POF group. By indirect immunofluorescence, only one patient was positive for antiovarian and antiadrenal antibodies. In summary, the results of this study are consistent with previous immunohistochemical data which indicate that autoimmune disorders are common in patients with POF. Furthermore, the detection of antiovarian antibodies in only one patient confirms that they are probably detectable in a short period of the disease.
Adolescent↗
Consequences of Fc gamma receptor type III reactivity in non-organ-specific autoimmune diseases.
Polymorphonuclear neutrophils (PMNs) express constitutively the Fc gamma receptors (Fc gamma Rs) Fc gamma RIIIb and Fc gamma RIIa for the Fc part of IgG, and soluble Fc gamma RIIIb (sFc gamma RIIIb) has been identified. Elevated levels of sFc gamma RIIIb were detected in non-organ-specific autoimmune conditions, and there was a considerably decreased number of PMNs undergoing apoptosis in the presence of sFc gamma RIIIb. Anti-Fc gamma RIIIb autoantibodies (autoAbs) have also been described in such patients. They are not cytotoxic to these cells, but they extend their survival. The anti-apoptotic signal can be transduced through Fc gamma RIIa and/or CD11b, the beta-chain of the complement receptor 3. However, Fc gamma RIIIb appears to be also competent. Anti-Fc gamma RIIIb-conditioned supernatant from cultured PMNs induces the transcription of messenger RNA for granulocyte colony-stimulating factor and granulocyte/macrophage colony-stimulating factor, followed by protein synthesis. The delay in apoptosis may be generated by a downregulation of the death promoter Bax. Inflammation might thus be modulated by sFc gamma RIIIb and anti-Fc gamma RIIIb autoAbs in systemic diseases.
Antigens, CD↗
Scoring and identifying organism-specific functional patterns and putative phosphorylation sites in protein sequences using mutual information.
The co-variance of amino acid positions within a multiple alignment of 294 protein kinases from mammals, plants, and bacteria was studied. Applying mutual information (MI), characteristic amino acid sites have been identified markedly discriminating the different organisms. The relation of surface accessibility of these sites in the 3D structure of a kinase and their MI content is studied. We extended the method to score a predicted phosphorylation site of this highly conserved catalytic protein kinase region. Based on this score mammalian and plant protein kinases were grouped together apart from the bacterial kinases. Thus, the presented method allows us to analyse putative phosphorylation sites in the context of their organism-specific origin.
Amino Acid Sequence↗
Platelet count and sepsis in very low birth weight neonates: is there an organism-specific response?
OBJECTIVE: Thrombocytopenia is commonly observed in very low birth weight (VLBW) neonates with sepsis. Specific platelet responses to different infectious agents have not been extensively characterized. The objectives of this study were to examine platelet counts and platelet indices in preterm neonates with culture-proven sepsis to determine if there are organism-specific platelet responses. STUDY DESIGN: We analyzed a cohort of all VLBW neonates (birthweight <1500 g) born over a 4-year period and admitted to a single level III neonatal intensive care unit (N = 943). Thrombocytopenia was defined as a platelet count <100,000/mm(3). Platelet count, nadir, duration of thrombocytopenia, and mean platelet volume (MPV) were examined during episodes of culture-proven sepsis. Analysis of variance, Kruskal-Wallis, Mann-Whitney U, and chi(2) tests were used to compare groups, and data are expressed as mean +/- standard deviation. RESULTS: Sepsis was diagnosed in 154 (16%) of 943 patients in the study population. Of the sepsis episodes, 54% were associated with thrombocytopenia and 61% with an elevation in MPV. Infections were grouped by organism type: Gram-positive bacteria (117/154, 76%), Gram-negative bacteria (24/154, 16%), and fungi (13/154, 8%). When compared with patients with Gram-positive sepsis, those with Gram-negative or fungal sepsis had a significantly lower initial platelet count, a lower platelet nadir, a higher incidence of thrombocytopenia, and a greater duration of thrombocytopenia. The decrease in platelet count from baseline was also significantly less in the Gram-positive infections than in the fungal infections. Although there was an overall increase in MPV from baseline, there were no differences between groups. CONCLUSIONS: In our population of VLBW infants, sepsis is frequently associated with thrombocytopenia and an elevation in MPV. However, fungal and Gram-negative pathogens are associated with a lower platelet count and more prolonged thrombocytopenia compared with Gram-positive pathogens. We conclude that common pathogens causing sepsis have different effects on platelet kinetics.
Adult↗