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PubMed2004

Highly sensitive assay for detection of enterovirus in clinical specimens by reverse transcription-PCR with an armored RNA internal control.

The objective of the present study was the development of a diagnostic reverse transcription (RT)-PCR for the specific detection of enterovirus (EV) RNA in clinical specimens controlled by an internal control (IC) RNA. The IC RNA contains the same primer binding sites as EV RNA but has a different probe region. The IC RNA was packaged into an MS2 phage core particle (armored) and was added to the clinical sample to allow monitoring of both extraction efficiency and RT-PCR efficiency. Serial dilutions of the IC RNA were made, and the detection limit of the RT-PCR was tested in a background of EV RNA-negative cerebrospinal fluid. The sensitivity and specificity of the RT-PCR assay were tested by using all 64 known EV serotypes, several non-EV serotypes, and two Quality Control for Molecular Diagnostics (QCMD) Program EV proficiency panels from 2001 and 2002. In total, 322 clinical specimens were tested by RT-PCR, and to establish the clinical utility of the RT-PCR, a comparison of the results of viral culture and RT-PCR was done with 87 clinical specimens. The lower limit of sensitivity was reached at about 150 copies of IC RNA/ml. All 64 EV serotypes were positive, while all non-EV serotypes were negative. All culture-positive samples of the 2001 QCMD proficiency panel (according to the 50% tissue culture infective doses per milliliter) were positive by RT-PCR. Invalid results, i.e., negativity for both EV RNA and IC RNA, due to inhibition of RT-PCR were observed for 33.3% of the members of the 2002 QCMD proficiency panel and 3.1% of the clinical specimens. Inhibition of RT-PCR could be relieved by the addition of 400 ng of bovine alpha-casein per microl to both the RT reaction mixture and the PCR mixture. With this optimized protocol, the results for all samples of the 2002 QCMD proficiency panel and all clinical specimens except one fecal sample (0.3%) were valid. Evaluation of the clinical samples demonstrated that EV infection could be detected in 12 of 87 samples (13.8%) by RT-PCR, while viral culture was negative. Our data show that the RT-PCR with armored IC RNA offers a very reliable and rapid diagnostic tool for the detection of EV in clinical specimens and that the addition of bovine alpha-casein relieved inhibition of the RT-PCR for 99.7% of clinical specimens.

Enterovirus↗
PubMed1983

A high molecular weight form of somatostatin-28 (1-12)-like immunoreactive substance without somatostatin-14 immunoreactivity in the rat pancreas. Evidence that somatostatin-14 synthesis can occur independently of somatostatin-28.

Synthesis of somatostatin-14 (S-14) could occur through direct enzymatic processing of precursor somatostatin (prosomatostatin) or via sequential breakdown of prosomatostatin leads to somatostatin-28 (S-28) leads to S-14. If direct processing is important, it should theoretically generate S-14 and a molecule equivalent to prosomatostatin without the S-14 sequence. In an attempt to identify such a molecule, I characterized the molecular forms of S-28(1-12)-like immunoreactivity (S-28(1-12) LI) in the rat pancreas and compared the relative amounts of these forms with those of S-14-like immunoreactivity (S-14 LI). Pancreatic extracts were chromatographed on Sephadex G-50 and Sephadex G-75 columns (Pharmacia Fine Chemicals Inc., Piscataway, NJ) under denaturing conditions and immunoreactivity in the eluting fractions was analyzed by region-specific radioimmunoassays (RIAs). For RIA of S-28(1-12) LI we used a newly developed rabbit antibody R 21 B, 125I-Tyr12 S-28(1-14), and S-28(1-12) standards. This system detects S-28, S-28(1-12), high molecular weight forms of S-28(1-12), but not S-14. S-14 LI was measured using antibody R149, which detects S-14, S-28, and higher molecular weight S-14-like substances, but not S-28(1-12). Three forms of S-28(1-12) LI were identified: Mr 9,000-11,000, Mr 1,200 (corresponding to S-28(1-12), and Mr less than 1,000, comprising, respectively, 35, 53, and 12% of total immunoreactivity. The relative abundance of the 9,000-11,000 mol wt S-28(1-12) LI material was unchanged following removal of S-14 LI from pancreatic extracts by affinity chromatography before gel filtration. Serial dilutions of fractions containing 9-11,000 and 1,200 mol wt materials exhibited parallelism with synthetic S-28(1-12). The total pancreatic concentration of S-28(1-12) LI was 1.56 pmol/mg protein, of which S-28(1-12) accounted for 0.83 pmol/mg protein and 9-11,000 S-28(1-12) LI comprised 0.55 pmol/mg protein. Pancreatic S-14 LI concentration was 2.07 pmol/mg protein, of which 98% corresponded to S-14. S-28-related peaks accounted for <1% of immunoreactivity in both RIAs. I concluded that (a) S-14 is the main form of pancreatic S-14 LI; (b) S-28 is present in very small quantities, in the pancreas; (c) S-28(1-12) LI consist mainly of S-28(1-12) and 9-11,000 mol wt S-28(1-12) LI; (d) 9-11,000 l wt S-28(1-12) LI could represent prosomatostatin without the S-14 sequence; (e) the finding of high concentrations of 9-11,000 mol wt S-28(1-12) LI suggests that S-14 synthesis can occur independently of S-28 and that direct processing of prosomatostatin is an important pathway for S-14 synthesis in the pancreas.

Animals↗
PubMed1993

Human plasma transport of vitamin D after its endogenous synthesis.

Transport of vitamin D3 from its sites of cutaneous synthesis into the circulation has been assumed to be via the plasma vitamin D binding protein (DBP). We studied vitamin D transport from the skin in seven healthy volunteers who received whole body irradiation with 27 mJ/cm2 dosage of ultraviolet B light (290-320 nm). Samples of venous blood were collected serially in EDTA and immediately chilled. In KBr, plasma samples were ultracentrifuged to provide a rapid separation of proteins of density < and > 1.3 g/ml. Upper and lower phases and serial fractions were analyzed for vitamin D3 (extraction, HPLC), cholesterol (enzyme assay), and human DBP (hDBP) (radial immunodiffusion). Total plasma vitamin D (basal level < 1 ng/ml) increased by 10 h and peaked at 24 h (9 +/- 1 ng/ml). 98% of the D3 remained at the density > 1.3 layers for up to 7 d, whereas cholesterol (> 85%) was detected at density < 1.3 and all of the hDBP was at density > 1.3. In three volunteers who each ingested 1.25 mg of vitamin D2, the total plasma D2 increased to 90 +/- 32 ng/ml by 4 h, and the D2 was evenly distributed between the upper and lower layers at 4, 8, and 24 h after the dose, indicating a continuing association of the vitamin with chylomicrons and lipoproteins, as well as with hDBP. Actin affinity chromatography removed D3 from plasma of irradiated subjects, indicating the association of the D3 with DBP. These findings indicate that endogenously synthesized vitamin D3 travels in plasma almost exclusively on DBP, providing for a slower hepatic delivery of the vitamin D and the more sustained increase in plasma 25-hydroxycholecalciferol observed after depot, parenteral administration of vitamin D. In contrast, the association of orally administered vitamin D with chylomicrons and lipoproteins allows for receptor-mediated, rapid hepatic delivery of vitamin D, and the reported rapid but less-sustained increases in plasma 25-hydroxycholecalciferol.

Adult↗
PubMed1991

Neuropeptide-Y in the trout brain and pituitary: localization, characterization, and action on gonadotropin release.

Using a specific antiserum raised against synthetic neuropeptide-Y (NPY), the distribution of NPY-like immunoreactivity in the brain and pituitary of the trout Oncorhynchus mykiss has been examined with the indirect immunofluorescence and peroxidase-antiperoxidase methods. The highest density of NPY-immunoreactive elements was found in the basal telencephalon and hypothalamus. In particular, NPY-immunoreactive neurons were located in the nucleus entopeduncularis and the preoptic nucleus. NPY-immunoreactive fibers were observed throughout the trout brain. The preoptic nucleus, the suprachiasmatic nucleus, and the nucleus entopeduncularis were densely innervated. In addition, NPY-positive fibers were detected in the nucleus lateralis tuberis and in the distal and intermediate lobes of the pituitary. The NPY-like peptide of the trout brain was characterized by combining HPLC analysis and radioimmunological detection. Serial dilutions of trout hypothalamus and pituitary extracts produced displacement curves that were parallel to the standard curve. HPLC analysis resolved a major peak which was slightly less hydrophobic than porcine NPY. The possible effect of NPY, either alone or in combination with a GnRH antagonist, on gonadotropin (GtH) release from trout pituitaries was investigated using a perifusion system technique. Graded concentrations of synthetic NPY induced a dose-dependent stimulation of GtH release. The stimulatory activities of NPY and various short chain analogs on GtH release were compared: the order of potency was NPY greater than NPY-(2-36) greater than NPY-(16-36) greater than NPY-(25-36). This result suggests that the biological determinant of NPY is located in the C-terminal part of the molecule. Administration of a short pulse of NPY or GnRH (10(-7) M each) induced a marked stimulation of GtH release. Prolonged infusion of the GnRH antagonist D-Phe2-6,Pro3-GnRH induced a significant reduction of GnRH-evoked GtH secretion. In addition, the GnRH antagonist blocked NPY-induced GtH release. The widespread distribution of NPY in the trout brain suggests the involvement of this neuropeptide in a variety of physiological functions. The present data support the view that NPY, released by nerve terminals in the distal lobe of the pituitary, may act presynaptically on GnRH fibers to modulate GtH release.

Animals↗
PubMed2004

Intraarterial pulmonary pentoxifylline improves cardiac performance and oxygen utilization after hemorrhagic shock: a novel resuscitation strategy.

UNLABELLED: The role of pentoxifylline (PTX) as a resuscitation adjunct in hemorrhagic shock is unclear. PTX infusion into the pulmonary artery and its effects on cardiac performance and oxygen utilization have not been defined. We hypothesized that pulmonary PTX is superior to systemic PTX or lactated Ringer's (LR) solution alone. The effects of LR solution, systemic PTX, and pulmonary PTX on cardiac performance and oxygen utilization in a hemorrhagic shock model in dogs were compared. Animals were bled to a mean arterial blood pressure (MAP) of 40 mm Hg maintained for 30 min and randomized into 3 resuscitation groups: LR solution (2x shed blood), systemic PTX (10 mg/kg bolus i.v.) in addition to LR solution (2x shed blood) + PTX (5 mg/kg for 45 min i.v.), and pulmonary PTX (10 mg/kg bolus + 5 mg/kg for 45 min via a pulmonary artery catheter) plus LR solution (2x shed blood, i.v.). Arterial blood gases, hemoglobin levels, MAP, cardiac index, systemic vascular resistance index, pulmonary vascular resistance index, oxygen delivery, oxygen consumption, and oxygen extraction ratio (O(2)ER) were measured serially. No differences in blood loss, hemoglobin, and MAP were observed. Pulmonary PTX increased cardiac index to levels more than baseline (P = 0.012) and decreased systemic vascular resistance index and pulmonary vascular resistance index to levels less than baseline (P < 0.0001). Pulmonary PTX increased oxygen delivery and oxygen consumption to baseline levels. Postresuscitation O(2)ER levels in LR-treated animals remained more than baseline (P < 0.0001). Systemic and pulmonary PTX significantly decreased O(2)ER compared with shock levels. PTX resuscitation is superior compared with LR solution alone. Intraarterial pulmonary PTX administration is safe, and improves cardiac performance as well as O(2) utilization. IMPLICATIONS: This study shows that a novel route (via the pulmonary circulation) used to administer pentoxifylline after hemorrhagic shock leads to superior cardiac performance in comparison with administration via lactated Ringer's solution or i.v. systemic pentoxifylline.

Animals↗
PubMed1988

Serial myocardial lactate metabolic changes after intracoronary thrombolysis in evolving myocardial infarction.

To investigate whether early reperfusion (less than or equal to 3 hours) preserves aerobic myocardial metabolism in acute myocardial infarction, we examined serial changes in trans-cardiac lactate extraction after thrombolysis in 43 patients with acute antero-septal myocardial infarction. In the chronic phase, we also determined abnormally contracting segments as an index of infarct size and regional ejection fraction as an index of chronic regional cardiac function. In the early reperfusion group (less than or equal to 3 hours), positive lactate extraction was restored and there were small abnormally contracting segments and a high regional ejection fraction. The intermediate reperfusion group (3-5 hours), however, had sustained anaerobic lactate extraction, large abnormally contracting segments and a low regional ejection fraction. The late reperfusion (greater than 5 hours) group showed apparently aerobic lactate extraction, but had large abnormally contracting segments and a low regional ejection fraction. Thus, early reperfusion preserves aerobic lactate metabolism and good chronic ventricular function.

Adult↗
PubMed1983

The effects of two strains of influenza virus on cardiac development in the chick embryo.

To investigate the teratogenic effects of influenza virus on cardiac development two strains of influenza virus (Ty/Calif/5142/66 and Ty/Calif/64/M. Meleagrium) were injected into the yolk sac of fertile white Leghorn eggs during stage 23. Eggs injected with normal saline served as controls. The number of live embryos was significantly reduced in eggs injected with the 5142 strain, whereas the Meleagrium strain did not affect the number of live embryos in a significant manner. Serial sections of the live embryo hearts, extracted during stage 35, showed no cardiac anomalies in control groups. However, cardiac malformations were increased significantly in the embryos infected with the influenza viruses. A comparison of the two viral strains showed that the 5142 strain produced significantly more abnormal hearts than the Meleagrium strain. We conclude that (1) influenza infection during critical stages of embryonic development can lead to the cardiac anomalies, and (2) the virulence of a viral strain has a significant bearing on its teratogenic potential, at least in chick embryo hearts.

Animals↗
PubMed2006

Studying multiple protein profiles over time to assess biomarker validity.

Protein profile analysis is increasingly used for identification of disease biomarkers. The approaches vary from surface-enhanced laser desorption/ionization to protein arrays. Newer platforms are constantly being developed. Almost all are based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and are often coupled with sophisticated software tools. Protein profiling has been applied to a variety of samples including plasma, urine, cerebrospinal fluid, saliva and solid tissue. This article focuses on those instances where it is possible to obtain sequential samples from the same individual. In the authors use of a profile method, many protein changes with highly significant correlations to disease have been found. The main challenge lies in the validation of the marker to demonstrate its adequacy for use in the clinical setting. The latter requires a methodology that is robust and amenable to high-throughput. One problem is that interindividual variability among the healthy population can mask major changes that occur on an intraindividual basis. Often, a large change for an individual may remain within the range of healthy individuals. Thus, one strategy to optimize biomarker discovery is to examine serial samples from a given individual, where a disease biomarker is established by comparison with the individual's own baseline sample. The focus of this review is to illustrate the principle and value of serial protein profiling using a rapid protein extraction method.

Adult↗
PubMed1998

Recombinant somatolactin as a stable and bioactive protein in a cell culture bioassay: development and validation of a sensitive and reproducible radioimmunoassay.

A recombinant somatolactin (SL) obtained by cloning and expression of sole SL cDNA was analyzed and used to develop a sensitive and specific RIA. In contrast to native proteins, which tend to dimerize and aggregate immediately after pituitary isolation, the majority of recombinant sole SL (rsSL) remained as a monomeric protein after long-term storage, as shown by size exclusion chromatography and Western blot. Using rsSL as a tracer and standard in the RIA, the minimum detectable dose and the midrange (ED50) of the assay were 0.15 and 1.8-2.1 ng/ml respectively. Intra-and interassay coefficients of variation were 4.3% and 6.5% at ED50 levels. Recombinant gilthead sea bream GH and recombinant trout GH did not show cross-reactivity, whereas a good parallelism between rsSL standard and serial dilutions of plasma and sole pituitary extracts was observed. In order to demonstrate some biological activity of rsSL, the ability of this recombinant product to prime gilthead sea bream phagocytes for in vitro enhancement of mitochondrial activity was examined by a chromogenic assay. A bell-shape dose-response curve was obtained with a maximum at 50 nM (1.2 micrograms/ml), similar to that reported previously for GH. Therefore, taking together all these data, it appears conclusive that rsSL is a long-term stable protein which retains, at least in part, biological activity, providing a useful tool to clarify the physiological role of fish SL.

Animals↗
PubMedDate not supplied

Detection of Cryptosporidium parvum in secondary effluents using a most probable number-polymerase chain reaction assay.

Polymerase chain reaction (PCR) was used to detect Cryptosporidium parvum oocysts in secondary effluent samples collected from activated-sludge facilities. Serial dilutions of the purified nucleic acid extracts from the samples were made and PCR was conducted to estimate the C. parvum oocyst concentration via a Poisson distribution-based most probable number (MPN). The degree of oocysts associated with wastewater particles was also evaluated. The sensitivity of the MPN-PCR assay was 20 oocysts/PCR unit. The detection limit of the concentration, extraction, and purification protocols in phosphate buffer saline spiked with a known concentration of oocysts ranged from 1.1 to 4.6 oocysts/L; the detection limit for the wastewater samples ranged from 11 to 4200 oocysts/L depending on the extent of inhibition in each sample. The recovery efficiency of the oocysts ranged from 48 to 59% in most samples. Oocysts were found in two out of seven samples with concentrations of 203 and 308 oocysts/L, as estimated by the MPN-PCR method. The oocysts were found only in the filtrate of the grab samples; particle-associated oocysts were not detected. Association of spiked C. parvum oocysts with particles in secondary effluent drawn from wastewater plants with varying operating conditions indicated a weak correlation between the degree of association and the mean cell residence time of the system.

Animals↗
PubMed2004

In vivo caries formation in enamel following argon laser irradiation and combined fluoride and argon laser treatment: a clinical pilot study.

OBJECTIVE: This in vivo pilot study investigated the role of argon laser irradiation and combined fluoride and argon laser treatment in accelerated natural caries development in sound enamel surfaces beneath plaque-retentive orthodontic bands. METHOD AND MATERIALS: Five patients (3 female, 2 male, ages 19 to 28 years) requiring tooth extraction prior to orthodontic treatment, participated in the study. Buccal surfaces were treated with either: (1) argon laser (250 mW for 10 seconds, ARGO-MOD); (2) topical fluoride (0.5% fluoride ion, Thera-Flur-N) followed by argon lasing; or (3) no treatment (control). Orthodontic bands with plaque-retentive slots on buccal surfaces were placed on the teeth slated for extraction (n = 14). Following a minimum of 5 weeks of intraoral exposure, the teeth were extracted for laboratory analysis. The teeth underwent serial longitudinal sectioning (12 sections per tooth). The sections were imbibed in water, and lesion depths were determined with each section, using polarized light microscopy. Comparisons were made among treatment groups (analysis of variance, Duncan's multiple range test for paired samples). RESULTS: Mean lesion depths were: 261 +/- 24 microm for the no treatment control group (n = 84 sections); 147 +/- 18 microm for the argon laser group (n = 24 sections); and 99 +/- 12 microm for the fluoride and argon laser group (n = 60 sections). Both the argon laser (44%) and the fluoride and argon laser groups (62%) had significant lesion depth reductions compared to controls. The addition of fluoride treatment prior to argon lasing resulted in a 32% reduction in lesion depth compared to argon laser treatment alone. CONCLUSIONS: Within this clinical pilot study, in vivo natural caries formation was affected significantly by a single exposure to low fluence argon laser irradiation. Topical fluoride treatment in combination with argon lasing provided an even greater degree of resistance against in vivo enamel caries development. A simple technique for reducing the caries susceptibility of enamel may be a clinical reality.

Adult↗
PubMed2005

[The creation of laosun yutie plaster and safety experiment].

OBJECTIVE: To create an externally-applied TCD plaster (Laosun Yutie plaster) for the lumbar muscle strain (LMS). METHOD: (1) The raw material such as angelica, safflower, frankincense, myrrh, clove and so on were turned into solid TCD plaster by a serial of modern treatment technology-distillation, extraction, condense, dehydration. Meanwhile, the naphtha from the distillation process and borneol were dissolved into ethanol solution and the dilution was thus made; (2) The solid TCD plaster and the dilution from the aboved-mentioned process were put together to form the TCD plaster and the corresponding safety experiments were applied to rabits' skin. RESULT: The experimental results showed that it is neither of urgent toxicity, stimulary function, nor of allergy-inducing response. CONCLUSION: The laosun Yutie plaster was a newly externally-applied TCD plaster which could be used safely and handily.

Administration, Cutaneous↗
PubMed1989

[Serial myocardial lactate metabolic changes after intracoronary thrombolysis in evolving myocardial infarction].

To ascertain whether early intracoronary reperfusion (less than 3 hours) preserves aerobic myocardial metabolism in acute myocardial infarction, serial changes in trans-cardiac lactate extraction after intracoronary thrombolysis were examined in 35 patients with acute anteroseptal myocardial infarction. Eight patients without intracoronary reperfusion served as controls. In the chronic phase, we also observed abnormally contracting myocardial segments as an index of infarct size and the regional ejection fraction as an index of chronic regional cardiac function. In the early reperfusion group (less than 3 hours; 15 cases), positive lactate extraction was restored; there were small abnormally-contracting segments and a high regional ejection fraction. However, the intermediate reperfusion group (3-5 hours; 10 cases) had sustained anaerobic lactate extraction, large abnormally-contracting segments and a low regional ejection fraction. The late reperfusion (greater than 5 hours; 10 cases) group showed apparent aerobic lactate extraction, but had large abnormally-contracting segments and a low regional ejection fraction. Thus, early reperfusion preserves aerobic lactate metabolism and good ventricular function in the chronic phase.

Heart Ventricles↗
PubMedDate not supplied

Patterns of reactivity of four novel monoclonal antibodies (B72.3, DF3, B1.1 and B6.2) with cells in human malignant and benign effusions.

The patterns of immunocytochemical reactivity produced in malignant and benign cells in effusions by staining with the monoclonal antibodies B72.3, DF3, B1.1 and B6.2 are reported. Using the mouse hybridoma methodology, these antibodies were raised against immunogens prepared from membrane-enriched extracts of metastatic carcinoma of the breast. Serial sections from the cell blocks of 20 benign effusions and 59 malignant effusions were incubated with the monoclonal antibodies using the avidin-biotin immunoperoxidase technique. The malignant neoplasms studied included specimens of 18 breast carcinomas, 12 ovarian carcinomas, 14 lung carcinomas, 7 leukemias/lymphomas and 8 miscellaneous malignant neoplasms. The monoclonal antibody B72.3 showed a positive staining reactivity with 36 of 39 adenocarcinomas (92%) versus staining in only 1 of 12 other neoplasms (8%). In contrast to the staining of adenocarcinomas, B72.3 stained no mesothelial cells in a total of 68 specimens tested. The monoclonal antibody DF3 showed a reaction pattern to neoplasms similar to that of B72.3 but also stained mesothelial cells in all of the specimens. The monoclonal antibodies B1.1 and B6.2 exhibited staining characteristics that have been associated with antibodies to carcinoembryonic antigen. The results of this study indicate that the monoclonal antibody B72.3 may function as a highly selective marker in recognizing a cancer cell versus a mesothelial cell and an adenocarcinoma cell versus other malignant tumor cells.

Adenocarcinoma↗
PubMed1993

Periodontal neural endings intimately relate to epithelial rests of Malassez in humans. A light and electron microscope study.

The periodontal ligament was examined by light microscopy at 3 different levels (apical, intermediate, coronal) on the 4 root sides of 43 extracted teeth. Epithelial rests were localised and serial LM and EM sections showed a close apposition (up to 0.03 microns) between Ruffini-like and free nerve endings and the basal lamina of the epithelial cell rests. The neural structures were facing the epithelial cells, whereas the Schwann cells were oriented towards the outer connective tissue. The Ruffini-like corpuscles contained numerous mitochondria. Free nerve endings contained neurotubules, neurofilaments and some vesicles. The intimate association between both neuroectodermal structures could indicate a target function of epithelial cell rests during developmental periodontal ligament innervation. Recent immunohistological findings involving nerve growth factor receptors substantiate this hypothesis.

Adolescent↗
PubMedDate not supplied

Sensitization to Olea europaea: geographical differences and discrepancies.

Thirty eight patients from two geographical areas of Spain, with great differences in Olea europaea pollen counts were studied to investigate their in vivo and in vitro immune response to this pollen as a consequence of their different environmental allergen exposure. They were distributed in two groups (13 from Madrid, and 25 from Jaén). Skin sensitivity was assessed by a prick-test dose-response bioassay using serial dilutions of a biologically standardized allergen extract of O. europaea. Serological immune response was evaluated measuring specific antibody levels (IgE, IgG, IgG1 and IgG4). The patients from Jaén, who have a higher exposure to olive pollen, had higher levels of specific antibodies but significantly smaller wheal sizes than a similar patient population form the Madrid area, where olive pollen is not so copious. There is a great discrepancy between the results of skin prick tests (low cutaneous reactivity associated with high allergenic environmental load) and the levels of specific IgE to the olive pollen. While the level of specific antibodies increases with the allergenic load, the capacity to release mediators seems to be decreased, at least in the skin. Further studies are needed to evaluate if these findings also occur in other target organs with appropriate challenge tests (conjunctival, nasal and bronchial). This pattern should be studied with other allergens in large patient populations.

Air↗
PubMed1999

Increased lipocortin-1 (annexin-1) expression in the sciatic nerve of Lewis rats with experimental autoimmune neuritis.

Lipocortin-1 exerts a potent immunosuppressive effect on pathogenic T cells. In multiple sclerosis and experimental autoimmune encephalomyelitis levels of lipocortins are raised, suggesting their involvement in the recovery from an immunological insult or in neural regeneration. To further understand the role of lipocortins in the peripheral nervous system we have characterized lipocortin-1 levels and cellular distribution of lipocortin-1 immunoreactivity in sciatic nerves of rats with experimental autoimmune neuritis (EAN), a model of human Guillain-Barré syndrome. EAN was induced actively by immunization with bovine peripheral myelin (active EAN) or by adoptive-transfer (AT-EAN) of P2-specific T cells. Cellular infiltrates in serial and semithin cryosections were characterized by immunohistochemistry. In parallel, lipocortin-1 levels in tissue extracts were quantified by a sandwich-ELISA. Only weak lipocortin-1 immunoreactivity was found in nerves of control animals injected with non-pathogenic T cells. The majority of macrophages and lymphocytes in EAN lesions exhibited lipocortin-1 immunoreactivity. Some very heavily stained cells showed a distribution and morphology similar to ED-2-positive macrophages which were abundant during early stages of EAN. Lipocortin-1 expression in T cells and macrophages was proven by immunocytochemical studies in semithin serial sections. In tissue extracts, lipocortin-1 levels increased from 0.24 +/- 0.14 micrograms/mg protein in controls receiving non-pathogenic T cells to a maximum of 0.55 +/- 0.1 micrograms/mg protein in AT-EAN at the peak of disease, and then slowly decreased during clinical recovery but still remained elevated. In dose-response studies in AT-EAN, highest values of lipocortin-1 (0. 71 +/- 0.23 micrograms/mg protein) were recorded after transfer of 2 x 10(7) T cells. Increased levels of lipocortin-1 were also measured in active EAN but occurred during the recovery phase (0.65 +/- 0.27 micrograms/mg protein). By analogy with other immune-mediated disorders, increased lipocortin-1 expression in the inflamed sciatic nerve in EAN may exert immunoregulatory functions in-situ and contribute to the termination of the autoimmune response.

Adrenal Cortex Hormones↗
PubMed2001

[Effects of 33% grapefruit extract on the growth of the yeast--like fungi, dermatopytes and moulds].

Grapefruit seed extract was discovered by Jacob Harich an american immunologist in 1980. Assessment of the influence of grapefruit extract on the yeast-like fungi strains--Candida albicans growth. Material used in this investigation was ATCC test Candida albicans strains no 10231, 200 of Candida albicans strains, 5 of Candida sp. strains isolated from patients with candidiasis symptoms from different ontocenosis and 12 of dermatophytes and moulds isolated from patients. The susceptibility of the Candida was determined by serial dilution method. It seems that 33% grapefruit extract exert a potent antifungal activity against the yeast like fungi strains and had low activity against dermatophytes and moulds. Further studies in vitro and in vivo on greater number of the yeast-like fungi strains and other fungi species are needed.

Arthrodermataceae↗