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At least 379 records · Page 21Linked to original sources
PubMed1975

Immunoelectron microscopic identification and localization of Streptococcus sanguis with peroxidase-labeled antibody: localization of surface antigens in pure cultures.

An indirect method of localizing antigens with horseradish peroxidase-labeled antibody was used to identify and localize surface antigens of Streptococcus sanguis at the ultrastructural level. An electron dense layer surrounding the cell wall could be distinguished without any additional electron microscope staining. This labeled layer represents an immune complex consisting of bacterial surface antigens, specific rabbit antisera, and peroxidase-labeled goat anti-rabbit globulins. Although with undiluted antisera slight cross-reactions occurred with S. salivarius and S. miteor (mitis), these could be readily distinguished from the more intense homologous reaction by their patchiness and the difference in distribution of the label. These cross-reactions were eliminated by appropriate dilutions of the antiserum. No cross-reactions occurred with S. mutans, S. faecalis, Actinomyces species, or Bacterionema, microorganisms wents indicated that horseradish peroxidase can become non-specifically adsorbed to the membrane of certain bacterial cells. Appropriate controls must, therefore, be included for localization of membrane associated antigens with horseradish peroxidase.

Actinomyces↗
PubMed1975

Immunoelectron microscopic identification and localization of Streptococcus sanguis with peroxidase-labeled antibody: localization of Streptococcus sanguis in intact dental plaque.

Streptococcus sanguis has been localized ultrastructurally within intact dental plaque by means of an indirect technique which utilizes horseradish peroxidase-labeled antibody. The technique allows for complete diffusion of the reagents to all portions of the plaque specimens. Control procedures can be carried out on serial sections of plaque with a bacterial composition similar to that of the experimental specimen. The 30-mum-thick sections can be examined in the light microscope to localize areas specifically labeled with peroxidase prior to cutting ultra-thin sections for electron microscopy. This study demonstrated that specific bacteria can be localized within intact dental plaque. The results also indicated that S. sanguis grows in dental plaque as columnar shaped microcolonies perpendicular to the tooth surfaces. Growth appears to be by cell division rather than deposition of new cells at the surfaces. Despite their relatively good structural preservation, the cells in the deeper (older) layers of plaque appear to have lost some of their antigenic activity in comparison to the cells near the surface.

Animals↗
PubMed1996

Allele-specific suppression of a defective trans-Golgi network (TGN) localization signal in Kex2p identifies three genes involved in localization of TGN transmembrane proteins.

Kex2 protease (Kex2p) and Ste13 dipeptidyl aminopeptidase (Ste13p) are required in Saccharomyces cerevisiae for maturation of the alpha-mating factor in a late Golgi compartment, most likely the yeast trans-Golgi network (TGN). Previous studies identified a TGN localization signal (TLS) in the C-terminal cytosolic tail of Kex2p consisting of Tyr-713 and contextual sequences. Further analysis of the Kex2p TLS revealed similarity to the Ste13p TLS. Mutation of the Kex2p TLS results in transport of Kex2p to the vacuole by default. When expression of a GAL1 promoter-driven KEX2 gene is shut off in MAT(alpha) cells, the TGN becomes depleted of Kex2p, resulting in a gradual decline in mating competence which is greatly accelerated by TLS mutations. To identify the genes involved in localization of Kex2p, we isolated second-site suppressors of the rapid loss of mating competence observed upon shutting off expression of a TLS mutant form of Kex2p (Y713A). Seven of 58 suppressors were allele specific, suppressing point mutations at Tyr-713 but not deletions of the TLS or entire C-terminal cytosolic tail. By linkage analysis, the allele-specific suppressors defined three genetic loci, SOI1, S0I2, and S0I3. Pulse-chase analysis demonstrated that these suppressors increased net TGN retention of both Y713A Kex2p and a Ste13p-Pho8p fusion protein containing a point mutation in the Ste13p TLS. SOI1 suppressor alleles reduced the efficiency of localization of wild-type Kex2p to the TGN, implying an impaired ability to discriminate between the normal TLS and a mutant TLS. soi1 mutants also exhibited a recessive defect in vacuolar protein sorting. Suppressor alleles of S0I2 were dominant. These results suggest that the SOI1 and S0I2 genes encode regulators or components of the TLS recognition machinery.

Alleles↗
PubMed2006

Antitumor effects of systemic and local immunization with a CTL-directed peptide in combination with a local injection of OK-432.

PURPOSE: The accumulation of T cells into the tumor site is crucial for the elicitation of in vivo antitumor effects after cancer vaccination. In this study, we investigated the antitumor effects and associated mechanisms of action that were induced by systemic and local immunization with a CTL-directed peptide in combination with a peritumoral injection of a streptococcal preparation, OK-432. EXPERIMENTAL DESIGN AND RESULTS: The human SART3(315-323) peptide, which has the potential to induce human leukocyte antigen-A24-restricted CTLs, not only has the same amino acid sequence as the mouse SART3, but also has the capacity for binding to H-2K(d) molecules. Therefore, the SART3(315-323) peptide could be used as a tumor antigen-derived peptide in H-2(d) mice. Systemic immunization with the SART3(315-323) peptide and the subsequent peritumoral injection of both the SART3(315-323) peptide and OK-432 effectively induced peptide-specific and colon26 carcinoma-reactive CTLs in BALB/c mice. The combination therapy suppressed the growth of s.c. established colon26 carcinoma. The accumulation of both CD8(+) and CD4(+) T cells into the tumor site was more apparent in mice treated with the combination therapy than in those treated with other protocols. In addition, the level of IgG reactive to the administered SART3(315-323) peptide increased in mice that were treated with the combination therapy. CONCLUSION: These results indicate that antitumor effects could be efficiently induced by a combination therapy that included systemic and local immunization with a CTL-directed peptide together with a local injection of OK-432.

Amino Acid Sequence↗
PubMedDate not supplied

Local hyperthermia, radiation, and chemotherapy in locally advanced malignancies.

Tumor size is a significant prognostic variable for attaining complete regression (CR) with local hyperthermia (HT) and radiation therapy (RT). The addition of weekly chemotherapy was evaluated to improve the efficacy of thermoradiotherapy in poor-prognosis lesions (i.e. > or = 7 cm2 or > or = 14 cm3) which have an expected CR rate of approximately 30 +/- 8%. Patients were entered into a two-arm phase-II study: arm 1 = breast cancer (10 patients), ifosfamide (1.5 g/m2) + epirubicin (20 mg/m2) + HT + RT; arm 2 = sarcoma (7 patients) and head and neck cancer (9 patients), cisplatin (40 mg/m2) + HT + RT. Therapy encompassing 106 triple-modality sessions was generally well tolerated for both arms; 2 instances of grade-3 and 1 of grade-4 (arm 2) local toxicity (WHO criteria) were observed. There were 4 instances of grade-3 myelosuppression (arm 1). The CR rates for arms 1 and 2 were 70 and 19%, respectively, suggesting that the combination of ifosfamide/epirubicin/HT/RT deserves further investigation in the context of localized breast cancer.

Adult↗
PubMed2001

Local delivery of enoxaparin to decrease restenosis after stenting: results of initial multicenter trial: Polish-American Local Lovenox NIR Assessment study (The POLONIA study).

BACKGROUND: Enoxaparin inhibits smooth muscle cell proliferation in experimental models. Intimal hyperplasia has been found to be the principal cause of restenosis after coronary stent implantation. We sought to determine whether the intramural delivery of enoxaparin before stenting of de novo lesions decreases restenosis. METHODS AND RESULTS: One hundred patients who were undergoing stenting were randomly assigned to either local administration of enoxaparin during predilation with reduced systemic heparinization or stenting with standard, systemic heparinization. All patients were treated with the same type of stent (NIR). The primary study end point was late luminal loss. The secondary end points were major adverse cardiac events, target lesion revascularization, and angiographic restenosis at 6 months. Angiographic follow-up at 6 months was completed in all except 1 patient. Late luminal loss was reduced to 0.76+/-0.42 mm in the local enoxaparin delivery group versus 1. 07+/-0.49 mm in the systemic heparinization group (P:<0.001). Restenosis, using a binary definition, occurred in 10% of patients in the enoxaparin group and in 24% of patients in the systemic heparinization group (P:<0.05). Target lesion revascularization rates occurred in 8% of the enoxaparin group and 22% of the systemic heparinization group (P:<0.05). There were no deaths and no emergent CABGs were performed. The only subacute stent closure and non-Q-wave infarction occurred in a patient assigned to the systemic heparinization group. CONCLUSIONS: This is the first prospective randomized trial in which the local delivery of a drug, enoxaparin, resulted in significant reduction in late luminal loss and restenosis after stent implantation in de novo coronary lesions.

Anticoagulants↗
PubMed1990

Effect of different fixatives on immunocytochemical localization of HNK-1-reactive antigens in cerebellum: a method for differentiating the localization of the same carbohydrate epitope on proteins vs lipids.

Monoclonal antibody (MAb) HNK-1 recognizes a carbohydrate epitope present in certain glycolipids, glycoproteins, and proteoglycans. Five different fixation methods, together with biochemical analyses of the antigens, were evaluated to study immunocytochemical localization of this epitope in layers of adult rat cerebellum; 4% paraformaldehyde/0.5% cetylpyridinium chloride was found to be optimal for overall immunoreactivity, and the antigens were apparent in all cerebellar layers. To differentially localize HNK-1-reactive carbohydrate epitope on proteins vs lipids in cerebellar layers, we tested the effect of 0.2%, 2%, or 4% glutaraldehyde combined with 2% paraformaldehyde (GT/PF) on HNK-1 and other MAb-reactive protein and lipid antigens; 2% or 4% GT/PF significantly reduced or abolished immunoreactivity of MAb HNK-1 and 5F9 (reacting with microtubule-associated protein 2) with cerebellar proteins analyzed on Western blots, but did not decrease HNK-1 reactivity to lipid antigens on HPTLC blots. In cerebellar tissue sections, HNK-1 and 5F9 immunoreactivity was reduced after 2% or 4% GT/PF fixation. However, significant amounts of HNK-1 immunoreactivity remained in molecular layer and deep cerebellar nuclei. GT/PF fixation did not cause significant changes in immunoreactivity patterns of other carbohydrate lipid antigens, such as those that react with MAb A2B5, 7A, and WCC4. Therefore, carbohydrate epitope on lipids, as opposed to that on proteins, may be preferentially detectable by immunocytochemistry after fixation with 2% or 4% GT/PF. The selective localization of HNK-1-reactive carbohydrate in the molecular layer and deep cerebellar nuclei with 2% or 4% GT/PF fixation correlates well with the observed presence of HNK-1-reactive lipids in these areas but not in the granular layer and white matter, as determined by microdissection of the individual layers and biochemical analysis. The application of 2% or 4% GT/PF fixation as a general method for differentiating the same carbohydrate epitope on proteins vs lipids in immunocytochemistry for other tissues and other antibodies remains to be further evaluated.

Animals↗
PubMed2005

Local coordination verses systemic disregulation: complexities in leukocyte recruitment revealed by local and systemic activation of TLR4 in vivo.

The recruitment of leukocytes to a tissue is a critical step in the inflammatory response. Toll-like receptor 4 (TLR4) is an important receptor involved in the initiation of inflammatory responses. Administration of the ligand for TLR4, lipopolysaccharide, is often used to model inflammation--local responses to stimuli within a specific tissue and systemic responses such as those observed during endotoxic or septic shock. Here, we review work, which demonstrates that in response to local activation of TLR4, highly coordinated and multistep processes are initiated, ultimately resulting in the leukocyte's arrival at the inflamed tissue. In contrast, systemic activation of TLR4 results in nonspecific accumulation of leukocytes within the lung capillaries and liver sinusoids through mechanisms profoundly different than those involved in local tissue recruitment. Contrary to current dogma, leukocyte accumulation in the lung is dependent on endothelial rather than leukocyte activation. Finally, we discuss recent evidence suggesting that activation of leukocytes through TLR4, although still in the circulation, effectively paralyzes inflammatory cells, rendering them incapable of appropriate trafficking to inflamed tissues.

Animals↗
PubMed1987

Intracellular localization of the glucocorticoid receptor: evidence for cytoplasmic and nuclear localization.

Using a monospecific, monoclonal antibody against the glucocorticoid receptor (GR), an immunocytochemical study was performed to investigate the intracellular localization of GR both in the presence or absence of ligand. With all fixation methods tested (paraformaldehyde, acetic acid in ethanol, Bouin's fixative, and bensochinone in PBS), it was possible to obtain specific GR staining. Fixation with paraformaldehyde was chosen for further studies on the effect of permeabilization, using several concentrations of Triton X-100 or saponin. A rat Rueber hepatoma (H-4-II-E) and a human uterus carcinoma (NHIK 3025) cell line were used as well as cultured hepatocytes from normal rat. The accessibility of the different cell compartments after fixation and permeabilization was tested for by using antibodies against cellular constituents with known locations (i.e. core-nucleosome proteins and tubulin), in combination with the anti-GR antibody in double immunofluorescence staining experiments. The specific GR stain obtained with the indirect peroxidase antiperoxidase technique or with fluorescein isothiocyanate-labeled second antibodies was shown to be present both in the cytoplasm and in the nucleus. Staining of all cellular compartments was abolished (peroxidase antiperoxidase) or diminished (fluorescein isothiocyanate) if the monoclonal antibody was preincubated with a 90% pure GR preparation. These findings are in contrast to recently reported immunocytochemical studies, where a strict nuclear existence of the estrogen and progestin receptors has been reported. Consequently, generalizations with regard to steroid receptor localization cannot be made. Furthermore, an in vitro model is described, where the effect of dexamethasone administration upon the localization of receptor staining in H-4-II-E cells can be studied.

Animals↗
PubMed1977

Local glucose utilization in acute focal cerebral ischemia: local dysmetabolism and diaschisis.

By means of an autoradiographic technique employing 14C-2-deoxyglucose, abnormalities of local brain glucose utilization were studied 90 minutes following occlusion of the left middle cerebral and common carotid arteries in a series of pentobarbital-anesthetized cats. Sham-insulted control animals exhibited a normal pattern of regional glucose utilization. In animals with vascular occlusion, a zone of greatly suppressed glucose utilization occupied the caudate nucleus of the ischemic hemisphere, with variable extension, and was surrounded by a narrow rim of increased local brain glucose utilization, suggesting the occurrence of enhanced anaerobic glycolysis in the latter zones. The cerebral cortex, which was less constantly affected, showed alternating regions of increased and decreased glucose utilization. Quantitation of local brain glucose utilization values from the contralateral nonischemic hemisphere revealed a mild suppression of cortical glucose utilization relative to the control animals. This may be the metabolic equivalent of diaschisis.

Animals↗
PubMed2000

Local thrombus formation in the site of intravenous injection of chlorpromazine: possible colloid-osmotic lysis of the local endothelial cells.

Since amphiphilic drugs are known to interact with biomembranes, we investigated local vessel damage and thrombosis which might be brought about by intravenous dosing using chlorpromazine (CPZ) as a representative compound. CPZ-induced hemolysis was suppressed by an increase in sucrose concentration in the medium, characterizing this hemolysis to be colloid-osmotic lysis, which includes the enhancement of membrane phospholipid fluidity and consequent small pore formation in the membranes. This was supported by the observation that hemolysis by filipin, not featuring the stage of small pore formation, was not affected by sucrose. [14C]Glucose-entrapping liposomes were degraded by CPZ, and this degradation was enhanced by an increase in the intravesicle glucose concentration. These results indicated that the compound could induce colloid-osmotic lysis in erythrocytes and artificial membrane vesicles. CPZ also injured cultured porcine aortic endothelial cells (PAEC), as evidenced by lactate dehydrogenase (LDH) leakage. This injury was also suppressed by increase in sucrose concentration in the medium, suggesting that colloid-osmotic lysis again occurred. When rats were intravenously injected with CPZ, local endothelial cell (EC) injury and associated thrombus formation were observed, suggesting that CPZ's action was also evident in vivo. To our knowledge, this is the first finding which suggests that an intravenously dosed amphiphilic drug can injure local ECs based on a colloid-osmotic lysis mechanism leading to thrombosis.

Animals↗
PubMed1988

[Stimulatory effects of lisuride on local cerebral blood flow and local cerebral glucose utilization in rats].

Effects of lisuride, a central dopamine agonist of the ergot type, on local cerebral blood flow (LCBF) and local cerebral glucose utilization (LCGU) were studied using the autoradiographical 14C-iodoantipyrine and 3H-deoxyglucose method, respectively, in conscious rats. Lisuride significantly stimulated LCBF in the cerebellar gray matter. LCBF in some other regions including the cerebral cortex, caudate-putamen, thalamus, geniculate body and substantia nigra were also stimulated by lisuride. No changes were observed in the hippocampus, hypothalamus and white matter. A close correlation was observed between LCBF and LCGU. Region specificity of the stimulatory effect by lisuride on LCBF and LCGU was coincident. Lisuride shortened the lethal time in the anoxia model. In conclusion, lisuride stimulated not only glucose utilization but also blood flow in local regions of the brain.

Animals↗
PubMed2004

Dynamic changes in intranuclear and subcellular localizations of mouse Prrp/DAZAP1 during spermatogenesis: the necessity of the C-terminal proline-rich region for nuclear import and localization.

Mouse Prrp (mPrrp)/DAZAP1 is a mouse ortholog of Xenopus Prrp, which is involved in vegetal pole localization of Vg1 mRNA in oocytes and is highly expressed in the testis. The mouse protein has been reported to be a shuttling protein which localizes in the nucleus of pre-meiotic spermatogenic cells and round spermatids, and shifts its location into the cytoplasm in elongating spermatids, suggesting that mPrrp may be involved in mRNA transport as well as that of the Xenopus ortholog. We reexamined immunohistochemical analyses of mPrrp/DAZAP1 during spermatogenesis utilizing a newly established monoclonal antibody and reconfirmed it to be a shuttling protein. We also carried out new observations that included remarkable intranuclear movement during spermatogenesis. In addition, we found that a long amino acid stretch which spanned over the C-terminal half of the protein was required for the nuclear import. These observations demonstrated dynamic changes in subnuclear and subcellular localization which might reflect specific functions during spermatogenesis.

Active Transport, Cell Nucleus↗
PubMed1979

Arteriography and chemotherapy in localized trophoblastic disease, by means of local- (pelvic-) intraarterial infusion.

In spite of considerable progress in systemic chemotherapy in malignant throphoblastic disease, there remains a small group of patients who fail to respond to this type of therapy. Here the trophoblastic process may be localized by means of an arteriography, and treated by a local, intraarterial infusion of cytotoxic compounds in high concentration. The latter makes possible the treatment of processes localized in essential organs which are hard to remove, as well as in young women wishing to preserve their reproductive capacity. Our experiences is limited to one case of a young woman with chorioadenoma destruens in the uterus, resistant to systemic chemotherapy, yet successfully treated with pelvic, intraarterial infusion of cytotoxics.

Adult↗
PubMed2000

Somatodendritic localization of 5-HT1A and preterminal axonal localization of 5-HT1B serotonin receptors in adult rat brain.

The 5-HT1A and 5-HT1B receptors of serotonin play important roles as auto- and heteroreceptors controlling the release of serotonin itself and of other neurotransmitters/modulators in the central nervous system (CNS). To determine the precise localization of these receptors, we examined their respective cellular and subcellular distributions in the nucleus raphe dorsalis and hippocampal formation (5-HT1A) and in the globus pallidus and substantia nigra (5-HT1B), using light and electron microscopic immunocytochemistry with specific antibodies. Both immunogold and immunoperoxidase preembedding labelings were achieved. In the nucleus raphe dorsalis, 5-HT1A immunoreactivity was found exclusively on neuronal cell bodies and dendrites, and mostly along extrasynaptic portions of their plasma membrane. After immunogold labeling, the density of membrane-associated 5-HT1A receptors could be estimated to be at least 30-40 times that in the cytoplasm. In the hippocampal formation, the somata as well as dendrites of pyramidal and granule cells displayed 5-HT1A immunoreactivity, which was also prominent on the dendritic spines of pyramidal cells. In both substantia nigra and globus pallidus, 5-HT1B receptors were preferentially associated with the membrane of fine, unmyelinated, preterminal axons, and were not found on axon terminals. A selective localization to the cytoplasm of endothelial cells of microvessels was also observed. Because the 5-HT1A receptors are somatodendritic, they are ideally situated to mediate serotonin effects on neuronal firing, both as auto- and as heteroreceptors. The localization of 5-HT1B receptors to the membrane of preterminal axons suggests that they control transmitter release from nonserotonin as well as serotonin neurons by mediating serotonin effects on axonal conduction. The fact that these two receptor subtypes predominate at extrasynaptic and nonsynaptic sites provides further evidence for diffuse serotonin transmission in the CNS.

Animals↗
PubMedDate not supplied

Local conjunctival immunotherapy: the effect of dermatophagoides pteronyssinus local conjunctival immunotherapy on conjunctival provocation test in patients with allergic conjunctivitis.

BACKGROUND: We evaluated the effect of local conjunctival immunotherapy (LCIT) with standardized dermatophagoides pteronyssinus (Dp) extracts on antigen-specific conjunctival provocation test (CPT) in patients with allergic conjunctivitis in a double blind, placebo-controlled study. We use the CPT because in our experience is the more objective parameter to evaluate the sensitivity to allergens in this patients. METHODS: The patients were selected on the basis of symptoms, positive prick test, positive CPT and elevated serum and tears total and specific IgE. The CPT was assessed with increased dilution of Dp extracts instilled into the lower fornix. Conjunctival hyperemia, tearing, itching, burning and swelling of eyelids were scored according a 4-point rating scale. Patients were randomly assigned to 2 groups of 12. The first group was treated with Dp extracts and the second group with placebo during 6 months. A drop of diluted antigen was instilled in both eyes daily, in 2-fold increased concentrations, the first 10 AU/ml. The maintenance dose was 1,000 AU/ml or the maximal dose which did not provoke symptoms. The symptoms were controlled with oral and/or local antihistamines. We evaluated the CPT before and after the treatment. The patients did not receive antihistamines during the 15 previous days to carrying-out the CPT. RESULTS: Ten of the twelve patients of the active group complete the treatment. One of the patients dropped out of the study because experienced local reaction with a dose of 1,000 AU/ml and refused to continue with the treatment. Other patient was disqualified for failure to comply with the protocol. One patient, which experienced itching and tearing with a dose of 1,000 AU/ml, tolerate 100 AU/ml. We continue with this dose until the end of treatment. The remaining patients tolerate 1,000 AU/ml as maintenance dose. A significant difference was observed in the score of CPT between LCIT treated patients and placebo group after 6 month of LCIT. CONCLUSIONS: We propose LCIT as a useful alternative to traditional subcutaneous immunotherapy in patients with allergic conjunctivitis.

Adolescent↗
PubMed2001

Local level sustainability policies in the Baltic Sea area: Local Agenda 21 within the Union of the Baltic Cities network.

Local Agenda 21 (LA21) processes have 2 central goals. i) On the basis of some of the empirical evidence in this study, the primary goal is to improve democratic (environmental) policy-making processes in such a manner that a larger share of the population will be able to participate in planning and decision making and will also be able to understand the consequences of these decisions. ii) The LA21 processes seek to improve (at least indirectly) the broadly defined environmental situation locally in a manner that takes into account both the local and the global contexts. The first part of this article discusses the concept and methods of LA21 and sheds light on the different action areas that are central to the Baltic LA21 processes. In addition, the study will describe and display the LA21 situation within one network of cities, the Union of the Baltic Cities (UBC). Networking, including transfer of information, models and ideas, has been among the main tools for the diffusion of LA21 ideas especially into newly democratized societies. Finally, the article will conclude with an overall assessment of the LA21 situation on the Baltic rim.

Baltic States↗
PubMed2003

[A case of locally extended rectal carcinoma quickly responding to local intraarterial infusion therapy].

Chemotherapy combined with 5-FU and l-leucovorin has been reported to elicit a quick response in cases of lower rectal carcinoma. A 65-year-old woman developed pain in the right gluteal region due to locally extended rectal carcinoma, and was treated with intraarterial infusion therapy. The chemotherapy regimen was 5-FU (500 mg/body) and l-leucovorin (175 mg/body) administered over 3 hours once weekly to bilateral reservoirs through an infuser pump. After 2 sessions of the chemotherapy, the gluteal pain decreased, and after 3 sessions, CEA level was confirmed and the patient's QOL began to improve. Side effects of the intraarterial infusion chemotherapy were gluteal dermatitis, leg desensitization, and infection of the circumferential reservoir. Local intraarterial infusion chemotherapy can be safely performed on an outpatient basis, and appears to elicit quick response for locally extended advanced rectal carcinoma.

Adenocarcinoma↗