Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Serial Extraction”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,099 records · Page 61Linked to original sources
PubMed2005

Comparison of real-time PCR protocols for differential laboratory diagnosis of amebiasis.

Specific identification of Entamoeba spp. in clinical specimens is an important confirmatory diagnostic step in the management of patients who may be infected with Entamoeba histolytica, the species that causes clinical amebiasis. Distinct real-time PCR protocols have recently been published for identification of E. histolytica and differentiation from the morphologically identical nonpathogenic Entamoeba dispar. In this study, we compared three E. histolytica real-time PCR techniques published by December 2004. The limits of detection and efficiency of each real-time PCR assay were determined using DNA extracted from stool samples spiked with serially diluted cultured E. histolytica trophozoites. The ability of each assay to correctly distinguish E. histolytica from E. dispar was evaluated with DNA extracted from patients' stools and liver aspirates submitted for confirmatory diagnosis. Real-time PCR allowed quantitative analysis of the spiked stool samples, but major differences in detection limits and assay performance were observed among the evaluated tests. These results illustrate the usefulness of comparative evaluations of diagnostic assays.

Animals↗
PubMed1998

Eight years' experience with the use of a transcystic common bile duct duodenal double-lumen catheter for the treatment of choledocholithiasis.

BACKGROUND: We have previously reported an alternative technique for treatment of choledocholithiasis found at laparoscopic cholecystectomy (LC) that can be considered with selected patients. This study was undertaken to update our experience with this alternative technique, which makes serial postoperative cholangiograms possible and facilitates stone extraction by assuring access to the common bile duct so that a guidewire-assisted endoscopic retrograde sphincterotomy can be performed. METHODS: In the period between 1989 and 1997, prospective data were maintained on 1043 consecutive patients who underwent LC by a single surgeon. Fifty-two patients with abnormal cholangiograms were managed with a percutaneously placed double-lumen catheter threaded through the cystic duct and advanced into the duodenum. RESULTS: Five attempts failed, 3 because of failure to pass the catheter and 2 because of catheter dislodgement. Of the 47 remaining patients, 2 underwent intraoperative endoscopic sphincterotomies using this alternative technique and 45 had cholangiograms repeated at 10 to 14 days. Twenty-three had negative cholangiograms, thus avoiding further procedures or unnecessary sphincterotomies because of spontaneous stone passage or initial false-positive cholangiograms. The remaining 22 had positive cholangiograms. Eighteen ultimately underwent sphincterotomies with stone extraction using a guidewire placed through the catheter. The other 4 had negative cholangiograms after serial follow-up, presumably because of spontaneous stone passage. CONCLUSIONS: The use of a transcystic double-lumen catheter passed through the ampulla of Vater is an effective and safe alternative for the management of choledocholithiasis discovered during LC.

Adolescent↗
PubMed1998

Serial sonography and colour flow Doppler imaging following testicular and epididymal sperm extraction.

Percutaneous epididymal sperm aspiration (PESA), percutaneous testicular sperm aspiration (TESA) and testicular sperm extraction (TESE) are invasive procedures and their consequences on the testis have not been clearly defined. In order to relate the sonographic and colour Doppler flow changes to the clinical data, 14 patients with non-obstructive and six with obstructive azoospermia were examined by the same roentgenologist immediately before, at 5 days, 2 weeks, 2 and 6 months after the surgical procedure. Testicular volumes remained unchanged during the follow-up period in both the non-obstructive and obstructive groups. Of the non-obstructive group, focal testicular lesions were seen in 20 of the 26 testes (77%) 5 days after the procedure and in 54% by 6 months. Ten were hypoechoic, of which six converted to echogenic foci, three remained hypoechoic and one disappeared at 6 months. The other 10 were echogenic lesions, three of which were no longer visible at 6 months and the remainder were unchanged. In the obstructive azoospermic group, focal lesions were not found. Extratesticular abnormality consistent with haematoma was demonstrated in four non-obstructive cases, which disappeared at the 6 month examination, and in none of the obstructive azoospermic patients. Whether residual focal lesions in the testes have long-term effects remains to be evaluated. In the obstructive azoospermic group, the aspirations performed did not leave any sonographic abnormalities.

Epididymis↗
PubMedDate not supplied

Capillary electrophoresis of the major anions and cations in leaf extracts of woody species.

Capillary electrophoresis methods are described for the analysis of the major inorganic anions (nitrite, nitrate, chloride, sulphate, phosphate), organic acids (oxalate, malate, citrate, succinate) and inorganic cations (ammonium, potassium, sodium, calcium, magnesium) in leaf extracts. Analytical performance was validated for extracts from leaves of four sclerophyllous species: Eucalyptus globulus, E. cladocalyx, E. nitens and Pinus radiata. Inorganic anions and organic acids were analysed in a single run within 5 min using a background electrolyte of 2,6-pyridinedicarboxylic acid (20 mM) and cetyltrimethylammonium bromide (0.5 mM). Cations were analysed in a separate run also within 5 min using imidazole (10 mM) and 18-crown-6 (2 mM) as background electrolyte. Replicate injections were highly repeatable when the capillary was rinsed between runs with hydrochloric acid (0.25 M) and background electrolyte, but not when the acid rinse was omitted or replaced by a rinse with sodium hydroxide (0.25 M). Standard curves for all analytes were linear over the range of 0.05-1 mm. Standard curves constructed by serial dilution of a leaf extract were also highly linear, and this, combined with the excellent recovery of added solutes in a spike and recovery test, suggests quantification was unaffected by the complex matrix that is present in un-purified, hot water extracts of leaves. There were significant differences in concentrations of the major anions and cations between the species studied.

Acids↗
PubMed2003

Anti-protozoal efficacy of medicinal herb extracts against Toxoplasma gondii and Neospora caninum.

The purpose of this study was to determine whether alcohol extracts of herbs (Sophora flavescens Aiton, Sinomenium acutum (Thunb.) Rehder and E.H. Wilson, Pulsatilla koreana (Yabe ex Nakai) Nakai ex T. Mori, Ulmus macrocarpa Hance and Torilis japonica (Houtt.) DC.) from South Korea, possess in vitro anti-protozoal activity against cultures of Toxoplasma gondii and Neospora caninum. These herbs have been used as human anti-parasitics in Asian countries for many years. Alcohol extracts of these herbs were serially diluted to final concentrations ranging from 625 to 19.5 ng/ml in media and added to wells containing either T. gondii or N. caninum tachyzoites in equine dermal (ED) cells. Parasite growth inhibition was measured using 3H-uracil incorporation as compared to untreated controls. T. japonica inhibited T. gondii proliferation by 99.3, 95.5, 73.0 and 54.0% in the range from 156 to 19.5 ng/ml, and S. flavescens inhibited T. gondii proliferation by 98.7, 83.0 and 27.2% in the range from 156 to 39 ng/ml. T. japonica inhibited N. caninum proliferation by 97.8, 97.9, 85.3 and 46.4% in the range from 156 to 19.5 ng/ml. S. flavescens inhibited N. caninum proliferation by 98.6, 97.0, 69.5 and 14.0% in the range from 156 to 19.5 ng/ml. Toxicity to host cells was noted when concentrations of T. japonica and S. flavescens exceeded 625 ng/ml. The herb extracts from S. acutum, Pulsatilla koreana, and U. macrocarpa also showed toxicity at higher levels but did not achieve the same inhibition effects at the lower concentrations against T. gondii and N. caninum as T. japonica and S. flavescens.

Animals↗
PubMed1994

In vitro antibacterial screening of Cryptolepis sanguinolenta alkaloids.

The ethanol and aqueous crude extracts and five alkaloids isolated from the roots of Crytolepis sanguinolenta (Lindl.) Schlechter were screened for antibacterial activity against 7 reference strains by the twofold serial broth microdilution assay. The ethanol extract and the alkaloids cryptolepine and cryptoheptine inhibited the growth of all strains tested except that of Pseudomonas aeruginosa.

Alkaloids↗
PubMed1975

Patterns of LH and FSH release from perifused rat pituitaries in response to infusions of hypothalamic extract.

A continuous flow incubation (perufusion) system was developed in which the secretory responses of pools of hemipituitaries from adult male rats to hypothalamic extract (HE) were characterized by serial radioimmunoassay of LH and FSH in the effluent medium. There was in initial massive release of LH and FSH which, in the absence of HE, declined to low basal levels at a rate which depended on the flow rate. Thereafter, the baseline for LH continued to decline gradually while that for FSH was stable. The rate of LH and FSH release rose abruptly after addition of the HE to the medium and returned promptly to baseline after withdrawal of the HE. During continuous infusion of HE for five hours, LH secretion was maintained at a relatively constant, elevated level. The responses to repeated identical pulses of HE were highly reproducible. The variability between responses by any one pool of tissue was significantly less than between responses of separate pituitary pools including pools comprising right and left halves of the same glands. For any given pool of pituitaries of the relationships were linear between: 1) duration of HE pulses (concentration constant) and increases in LH output, and 2) log of concentration of HE (pulse duration constant) and increases inLH and in FSH output. Consistent responses were obtained for up to 12 hr, the maximum period tested.

Animals↗
PubMed1980

A controlled study of the effectiveness of the Rinkel method of immunotherapy for ragweed pollen hay fever.

In a double-blind study, we compared the effects of the Rinkel method of immunotherapy with ragweed pollen extract and placebo on symptoms of ragweed hay fever and immunologic parameters in 24 ragweed-sensitive patients. Each had a skin-test end point by Rinkel serial dilution titration to ragweed pollen extract at 1:312,500 w/v or greater dilution, a 2 + skin test to ragweed AgE at 0.1 microgram /ml or greater dilution, and in vitro leukocyte histamine release by ragweed pollen extract. None had had immunotherapy for at least 7 yr. Patients matched on the basis of leukocyte histamine release by ragweed were assigned to two treatment groups (12 patients in each group). One group received ragweed pollen extract, and the other, placebo, both administered by the Rinkel method between June and October, 1978. Treatment doses were derived from skin-test end points. The median maintenance ("optimal dose") for patients receiving ragweed pollen extract was 0.53 ml of 1:312,500 w/v and the mean cumulative dose of ragweed pollen extract given during the study contained 0.094 micrograms of ragweed AgE. Symptom-medication scores of all patients rose and fell with ragweed pollen counts. No significant differences were observed in mean daily symptom-medication scores, antiragweed IgG or IgE levels, leukocyte histamine release by ragweed, total IgE levels, or skin-test end-point dilutions with ragweed pollen extract between the group receiving ragweed pollen extract and the group receiving placebo. Despite the absence of specific effect on symptom-medication scores and measured immunologic variates, 10 3f the 12 ragweed-treated patients and 10 of the 12 placebo-treated patients were of the opinion that their hay fever symptoms during the ragweed pollen season were less severe in 1978 than in 1977 and that they had been helped by Rinkel method immunotherapy. Under the conditions of the study, Rinkel method immunotherapy with ragweed pollen extract was no more effective than placebo given in an imitation of the Rinkel method.

Adolescent↗
PubMed1995

The effects of smokeless tobacco extract on bone nodule formation and mineralization by chick osteoblasts in vitro.

Short-term exposure to smokeless tobacco extracts (STE) reportedly inhibits osteoblast metabolism. The objective of this study was to determine the effects of serial dilutions of a water-soluble extract of smokeless tobacco on osteoblast proliferation and their potential to form and mineralize bone nodules. STE significantly stimulated cell proliferation when diluted 10(2)-10(4) times; 10(3) and 10(4) dilutions produced the greatest effect. 10(2)-10(4) STE dilutions significantly increased alkaline phosphatase activity at day 7 but 10(6) STE dilutions significantly decreased it. 10(3) and 10(4) dilutions significantly increased bone nodule formation, but inhibited their mineralization. In contrast, 10(5) and 10(6) dilutions significantly decreased bone nodule formation, but increased their mineralization. Stimulation of in vitro bone nodule formation by STE was similar to that produced by 10(-7) M insulin-like growth factor 1 (IGF-1) in vivo. Heat and acid treatment of STE significantly reduced its beneficial effect on cell proliferation, suggesting that a peptide within STE may be responsible for enhancement of osteogenic cell proliferation. Thus, STE may contain a peptide capable of significantly stimulating osteoblast proliferation, differentiation and metabolism, similar to the effects of IGF-1. This peptide could have potential therapeutic benefits.

Analysis of Variance↗
PubMedDate not supplied

Antibacterial activity of vegetables and juices.

OBJECTIVE: We evaluated the antibacterial activities of various fruit and vegetable extracts on common potential pathogens including antibiotic-resistant strains. METHODS: Standardized bacterial inocula were added to serial dilutions of sterile vegetable and fruit extracts in broth, with final bacterial concentrations of 10(4-5) cells/mL. After overnight incubation at 35 degrees C, antibacterial activity was measured by minimum inhibitory and minimum bactericidal dilutions (for raw juices) or concentrations (for tea). RESULTS: Among the vegetable and fruit extracts tested, all green vegetables showed no antibacterial activity on Staphylococcus epidermidis and Klebsiella pneumoniae. All purple and red vegetable and fruit juices had antibacterial activities in dilutions ranging from 1:2 to 1:16. Garlic juice had significant activity, with bactericidal action in dilutions ranging up to 1:128 of the original juice. Tea also had significant activity, with bactericidal action in concentrations ranging up to 1.6 mg/mL, against a spectrum of pathogens including resistant strains such as methicillin- and ciprofloxacin-resistant staphylococci, vancomycin-resistant enterococci, and ciprofloxacin-resistant Pseudomonas aeruginosa. CONCLUSIONS: Tea and garlic have the potential for exploration of broader applications as antibacterial agents.

Anti-Bacterial Agents↗
PubMed2004

Effect of comprehensive validation of the template isolation procedure on the reliability of bacteraemia detection by a 16S rRNA gene PCR.

The influence of the DNA extraction method on the sensitivity and specificity of bacteraemia detection by a 16S rRNA gene PCR assay was investigated. The detection limit of the assay was 5 fg with purified DNA from Escherichia coli or Staphylococcus aureus, corresponding to one bacterial cell. However, with spiked blood samples, the detection limits were 10(4) and 10(6) CFU/mL, respectively. The sensitivity of the S. aureus assay was improved to the level of the E. coli test with the addition of proteinase K to the commercial DNA extraction kit protocol. Ten (16.6%) of 60 amplification reactions were positive with templates isolated from sterile blood, while PCR reagent controls were negative, thereby indicating contamination during the DNA extraction process. Blood samples were spiked with serial dilutions of E. coli and S. aureus cells, and six PCR results were obtained from three extractions for each blood sample. A classification threshold system was devised, based on the number of positive reactions for each sample. Samples were deemed positive if at least four positive reactions were recorded, making it possible to avoid false-positive results caused by contamination. These results indicate that a comprehensive validation procedure covering all aspects of the assay, including DNA extraction, can improve considerably the validity of PCR assays for bacteraemia, and is a prerequisite for the meaningful detection of bacteraemia by PCR in the clinical setting.

Bacteremia↗
PubMed2004

The risk of development of antimicrobial resistance in individual patients with chronic rhinosinusitis.

OBJECTIVE: To determine if individual patients with chronic rhinosinusitis (CRS) demonstrate an increasing prevalence of antimicrobial resistance over time. PATIENTS: A prospectively maintained database of microbiologic culture results for adult patients with CRS was sequentially analyzed, extracting patients who underwent multiple serial sinus cultures over time. Antimicrobial resistance patterns were identified and quantified for each bacterium. Sequential culture results were compared for individual patients to determine if increasing overall bacterial resistance, gram-positive resistance, gram-negative resistance, or a shift toward gram-negative organisms was manifested within individual patients. RESULTS: During a 7-year period, 90 adult patients were identified with 224 serial cultures (mean, 2.5 cultures per patient) obtained, with a median time between cultures of 157 days. Four hundred twenty-nine organisms were isolated from these serial cultures, consisting of 255 gram-positive organisms, 120 gram-negative organisms, 48 anaerobes, and 6 fungi. Pairwise analysis of sequential cultures revealed no significant trend toward increasing bacterial resistance within individual patients (P = .57, runs test). Similarly, no significant trend toward increasing gram-positive or gram-negative resistance was demonstrated. There was no shift toward gram-negative organisms (P>.15 for all). CONCLUSIONS: Individual patients with CRS do not necessarily develop increasing levels of bacterial resistance over time. The use of culture-directed antimicrobial therapy may "protect" against the development of sequentially increasing antimicrobial resistance for patients with CRS.

Adult↗
PubMed1994

Genotypes and titers of hepatitis C virus for predicting response to interferon in patients with chronic hepatitis C.

Interferon induces remission in about 50% of patients with chronic hepatitis C, but it is difficult to predict which patients will respond. Host and viral factors were evaluated for correlation with response to interferon in patients with chronic hepatitis C. Recombinant interferon alpha-2b with a total dose of 480-560 million units was given to 136 patients, of whom 74 (54%) responded. Genotypes of hepatitis C virus (HCV) in sera, I, II, III, IV, and V, were determined by polymerase chain reaction (PCR) with type-specific primers. In 72 patients, pretreatment levels of HCV RNA were titrated by PCR in serial tenfold dilutions of RNA extracted from serum. Response to interferon occurred in 34 (40%) of 85 patients infected with HCV of genotype II, less frequently than in 22 (85%) of 26 with genotype III (P < 0.001) or in 7 (70%) of 10 with genotype IV. Of 51 patients with genotype II HCV, 6 of 8 (75%) with HCV RNA titers < 10(6) responded, more frequently than 4 of 43 (9%) with titers > or = 10(6) (P < 0.001). Responders were younger than non-responders (45.7 +/- 11.7 vs. 50.3 +/- 9.6 yr) and had received transfusions less frequently (26/74 or 35% vs. 37/62 or 60%, P < 0.01). Response to interferon correlated inversely with the severity of liver histopathology. These results indicate that response to interferon is influenced by HCV genotypes and pretreatment levels of HCV RNA in serum.

Adult↗
PubMed1994

Monitoring of renal allograft recipients by quantitation of human cytomegalovirus genomes in peripheral blood leukocytes.

The ratio of human cytomegalovirus (HCMV) genomes per cellular genomes in serial peripheral blood leukocyte (PBL) extracts of renal allograft recipients was quantitated by competitive nested polymerase chain reaction (PCR). Patients were also monitored for the development of acute HCMV infection by detection of HCMV pp65 antigenemia, HCMV IgM antibodies, and viruria. Compared to qualitative nested HCMV PCR, the frequency of positive PCR results in renal allograft recipients without further evidence of acute HCMV infection was significantly reduced by quantitative HCMV PCR. HCMV DNA levels > or = 1,000 copies HCMV/10(6) copies beta-globin were found to be highly indicative for the development of a clinically symptomatic HCMV infection following renal allograft transplantation. In patients treated with ganciclovir, quantitation of HCMV target sequences allowed the assessment of the efficacy of antiviral therapy.

Antibodies, Viral↗
PubMed1994

Atrial natriuretic peptide immunoreactivity in the eggs of the silkworm Bombyx mori.

Synthesis and secretion of atrial natriuretic peptides (ANPs) are not confined to the atrium, but are also present in other tissues. Recently, we have found synthesis of ANP in the eggs of several vertebrate animals. The present study was undertaken to determine whether immunoreactive ANP (irANP) is present in the egg of an invertebrate, the silkworm (Bombyx mori L.). The serial dilution curve of egg extracts of silkworm was parallel to the standard curve of atriopeptin III. Analysis of ANP immunoreactivity by gel filtration chromatography and reverse-phase HPLC showed that the major immunoreactivity corresponded to rat proANP. The semipurified irANP of egg extracts produced a dose-dependent relaxation on rat aortic strips, which was blocked by preincubation with anti-ANP antiserum. Therefore, we suggest that ANP is synthesized in the silkworm egg.

Animals↗
PubMed1994

A radioimmunoassay for the determination of arginine vasotocin (AVT): plasma and pituitary concentrations in fresh- and seawater fish.

A specific radioimmunoassay was developed and characterized for the measurement of arginine vasotocin (AVT) in teleost fish. Specificity of the antibody for AVT was demonstrated by parallelism of a series of AVT standards with serially diluted pituitary and plasma extracts. Crossreactivity of the antibody with the other teleost neurohypophysial peptide, isotocin, was less than 1% and the sensitivity of the assay was 0.24 fmol/assay tube. AVT was extracted from plasma by reverse-phase liquid chromatography [efficiency of 87.6 +/- 9.3% (n = 5)] and demonstrated as an effective procedure for plasma volumes ranging from 0.4 to 1.2 ml. Plasma AVT concentrations measured in a range of euryhaline and stenohaline teleost fish were between 10(-12) and 2 x 10(-11) M (1-20 pg/ml). There were no consistent differences between plasma AVT levels in euryhaline fish (flounder, trout, and eel) adapted to fresh water (FW) and sea water (SW). In flounder, pituitary AVT levels in FW- and SW-adapted fish were also similar.

Adaptation, Physiological↗
PubMed1993

Small intestinal mucosa changes, including epithelial cell proliferative activity, of children receiving total parenteral nutrition (TPN).

We examined the small intestinal histology disaccharidase activities as well as the incorporation of [3H]thymidine into DNA of biopsies maintained in organ culture from seven children (ages 9 months to 5 years) receiving total parenteral nutrition (TPN). Three children suffered from inflammatory bowel disease and received TPN for one month (short term). Four required long-term TPN (> 9 months) for short-bowel syndrome. DNA was extracted from the samples following serial precipitation with perchloric acid. Results were compared to those from 22 age-matched children investigated for abdominal pain or chronic diarrhea. Short-term TPN resulted in slightly lower lactase, sucrase, and palatinase activities that were not statistically different from controls. Long-term TPN resulted in focal mild villus atrophy and a decrease in disaccharidase activity in two patients. Biopsies from long-term TPN patients incorporated less thymidine compared to those of controls (P < 0.001) when data was expressed per total biopsy (3.6 +/- 1.1 vs. 8.4 +/- 1.1 fmol) or per milligram of tissue (1.0 +/- 0.12 vs 2.7 +/- 0.7 fmol). The above data are in general agreement with the hypoplastic effect of TPN in animals. However, in children, much longer periods of TPN are required to realize the changes.

Adolescent↗