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At least 1,171 records · Page 65Linked to original sources
PubMed2006

Gene transfer of cytidine deaminase protects myelopoiesis from cytidine analogs in an in vivo murine transplant model.

Hematopoietic stem cell gene transfer of the drug-resistance gene cytidine deaminase (CDD) protecting cells from the cytotoxic cytidine analogs cytarabine and gemcitabine was investigated in a murine transplant model. Following transplantation of CDD-transduced cells and cytarabine application (500 mg/kg; days 1-4; intraperitoneally) significant myeloprotection was demonstrated with nadir counts of peripheral blood granulocytes and thrombocytes of 2.9 +/- 0.6/nL versus 0.7 +/- 0.1/nL (P < .001) and 509 +/- 147/nL versus 80 +/- 9/nL (P = .008), respectively (CDD versus control). Protection also was observed from otherwise lethal gemcitabine treatment (250 mg/kg; days 1-3). Stable levels of gene-marked cells in primary and secondary recipients were demonstrated for up to 9 months, and whereas CDD overexpression clearly reduced B- and T-lymphocyte numbers, no major toxicity was observed in the myeloid compartment. Despite the profound myeloprotective properties, however, CDD overexpression did not allow for pharmacologic enrichment of transduced hematopoiesis in our model. Thus, in summary, our data establish CDD as a drug-resistance gene highly suitable for myeloprotective purposes, which, given the lack of selection observed in our hands, might best be used in combination with selectable drug-resistance genes such as MGMT (P140K) or MDR1.

3T3 Cells↗
PubMed2000

Comparison of cardiac stroke volume measurement determined using stereological analysis of breath-hold cine MRI and phase contrast velocity mapping.

Cine MRI of the heart using segmented k-space pulse sequences permits multiphase images to be acquired in a single breath-hold. Whilst image quality is improved compared with conventional (non-segmented) cine imaging, subsequent analysis can be relatively time consuming. In this study, multiple slice breath-hold cine imaging of the heart was performed in 11 normal volunteers. Left and right ventricular stroke volume (SV) was estimated from the images by application of the Cavalieri method of modern design stereology in combination with point counting. The measured SVs were compared with those obtained by cine phase contrast velocity mapping in the ascending aorta and main pulmonary artery. Excellent agreement was found between the SVs determined by the two techniques, with mean differences (+/- one SD) of 0.41 +/- 3.00 ml and 0.41 +/- 4.78 ml for left and right ventricles, respectively. Comparison of left and right ventricular SV using stereology yielded a mean difference of 0.84 +/- 5.70 ml. Breath-hold data acquisition together with stereological analysis is demonstrated to be an accurate and unbiased technique for the rapid assessment of cardiac function.

Adult↗
PubMed1986

Estimation of chicken microbial load using the growth curve parameters of chicken microflora.

Chicken microbial loads, estimated through absorbance increase of culture medium inoculated with the contaminant microflora of the carcasses, were compared with total plate counts and psychrotrophic counts obtained on the same carcasses using the pour plate method after 0, 48, 96, and 144 hr stored chicken at 4 to 5 C. For estimating microbial loads on the carcasses, the mathematical relation 1n AO = 1nA1 - R - Kt was used, which was developed by combining the growth and R equations (described in Materials and Methods) and using growth data at 28 C. The values obtained by this method, when compared with those of plating, give correlation coefficients of .94, .91, .88, and .64 for total plate counts after 0, 48, 96, and 144 hr of storage and .94, .83, .82, and .86 for psychrotrophics counts after 0, 48, 96, and 144 hr of cold storage. The method proposed in the present work permits the estimation of psychrotrophics and total counts in no more than 11 hr, which is very promising for industrial applications.

Animals↗
PubMedDate not supplied

Immunomodulating and antimetastatic activity of thymic peptides in BALB/c mice.

The immunomodulating and antimetastatic activity of a clinically approved low molecular weight, standardized thymic peptide (TP) preparation was evaluated in BALB/c-mice. Daily subcutaneous application of TP (7 consecutive days, 1 and 10 micrograms per mouse) upregulated counts and activity of peripheral blood cells, as measured on day 14. To check the influence of TP on experimental liver metastases (liver colonization), RAW H10 lymphosarcoma cells were intravenously inoculated into BALB/c-mice. TP (1,10,100 micrograms/mouse) was subcutaneously administered daily for 7 consecutive days starting 24 hrs after tumor cell challenge. Liver colonization was investigated on day 14 after tumor cell inoculation and demonstrated a statistically significant (p < 0.05) reduction of experimental liver metastases for TP treated mice.

Adjuvants, Immunologic↗
PubMed2005

[Spectrometric assessment of thyroid depth within the radioiodine test].

UNLABELLED: Aim of this study is the validation of a simple method for evaluating the depth of the target volume within the radioiodine test by analyzing the emitted iodine-131 energy spectrum. PATIENTS, METHODS: In a total of 250 patients (102 with a solitary autonomous nodule, 66 with multifocal autonomy, 29 with disseminated autonomy, 46 with Graves' disease, 6 for reducing goiter volume and 1 with only partly resectable papillary thyroid carcinoma), simultaneous uptake measurements in the Compton scatter (210 +/- 110 keV) and photopeak (364-45/+55 keV) windows were performed over one minute 24 hours after application of the 3 MBq test dose, with subsequent calculation of the respective count ratios. Measurements with a water-filled plastic neck phantom were carried out to perceive the relationship between these quotients and the average source depth and to get a calibration curve for calculating the depth of the target volume in the 250 patients for comparison with the sonographic reference data. Another calibration curve was obtained by evaluating the results of 125 randomly selected patient measurements to calculate the source depth in the other half of the group. RESULTS: The phantom measurements revealed a highly significant correlation (r = 0,99) between the count ratios and the source depth. Using these calibration data, a good relationship (r = 0,81, average deviation 6 mm corresponding to 22%) between the spectrometric and the sonographic depths was obtained. When using the calibration curve resulting from the 125 patient measurements, the overage deviation in the other half of the group was only 3 mm (12%). There was no difference between the disease groups. CONCLUSION: The described method allows on easy to use depth correction of the uptake measurements providing good results.

Carcinoma, Papillary↗
PubMedDate not supplied

[Adaptation strategies of halophilic microorganisms and Debaryomyces hansenii (halophilic yeast)].

The term halophile is used for all those organisms belonging to hypersaline habitats; they constitute an interesting class of organisms able to compete successfully in salt water and to resist its denaturing effects. A wide diversity of microorganisms, prokaryotic and eukaryotic belong to this category. Halophile organisms have strategies allowing them not only to withstand osmotic stress, but also to function better in the presence of salt, in spite of maintaining high intracellular concentrations of salt, partly due to the synthesis of compatible solutes that allow them to balance their osmotic pressure. We describe the characteristics of some halophile organisms and D. hansenii (halophile yeast), that allow them to resist high concentrations of salt. The interest to know the great diversity microorganisms living in hypersaline habitats is growing, and has begun to be the center of recent investigations, since halophile organisms produce an wide variety of biomolecules that can be used for different applications. In this review we describe some mechanisms with which some halophile organisms count to resist the high concentration of salts, mainly NaCl.

Adaptation, Physiological↗
PubMed1990

A simple method for in-vitro studies of cancer tissue metabolism.

A simple laboratory method of investigating the metabolism of malignant tissues has been devised for liver cancer. The apparatus consists of a standard glass scintillation vial with an inner glass well suspended within it from a rubber lined cap. 14C-labelled glucose metabolised by the tumour tissue in the inner well into 14CO2 will be trapped by alkali in the outer vial, and the radioactivity quantified by liquid scintillation counting. The counts obtained will give a measure of the rate of tissue glucose oxidation. Application of this method to an animal liver tumour model has shown that the tumour tissue has a glucose oxidation rate of 2.8 times that of the host liver. The method is simple and versatile, can be adapted to various types of tissues, and may have potential clinical applications.

Animals↗
PubMed1980

Activation of antibacterial silver coatings on surgical implants by direct current: preliminary studies in rabbits.

Staphylococcus aureus contamination on intramedullary silver pins in the femurs of rabbits was decreased 69% by the application of 9 muA for 1 hour of positive direct current/cm2 (9muAH+DC/cm2). Injection of a solution of silver chloride or silver nitrate into the bone marrow before insertion of the contaminated pins and subsequent application of 12 muAH+DC/cm2 to the stainless steel pins reduced the colony counts 25% and 64%, respectively, and indicated a need for the Ag+ and Cl- ionic state on the pins' surfaces. Stainless steel pins electroplated with silver and charged with 12 muAH+DC/cm2 decreased the bacterial counts by 85%. A 91% inhibition with stainless steel pins electroplated with silver and chlorided indicated that bacteriostasis was possible due to the Ag+/AgCl/Cl- surface effect. Total bacterial inhibition was achieved by 12 muAH+DC/cm2 on stainless steel pins coated with 100 monomolecular layers of silver stearate. The electrical activation of this coating eliminated up to approximately 3,000 S aureau contaminating the intramedullary pins.

Animals↗
PubMed1994

Cervicovaginal screening in women with HIV infection: a need for increased vigilance?

OBJECTIVE: To review the current literature on cervical disease (dysplasia, cervical intraepithelial neoplasia [CIN] or carcinoma) in women with HIV infection and to assess recommendations for cervicovaginal screening in these patients. DATA SOURCES: MEDLINE and AIDSLINE were searched for relevant articles published in English or French between January 1987 and February 1993, abstracts presented at international AIDS conferences from 1989 to 1993 were evaluated, and pertinent agencies and organizations were consulted. STUDY SELECTION: A total of 92 reports of gynecologic disease in women with HIV infection were examined; 32 studies were retained that reported pertinent findings on cervical dysplasia, CIN or cervical carcinoma. DATA EXTRACTION: The following criteria were used to extract data: study design (descriptive v. comparative), sample size, heterogeneity of the study population, presence of immunodeficiency indicators (i.e., absolute CD4+ lymphocyte count) and presence of concomitant vaginal infections. Recommendations were assessed for their specific application to women with HIV infection. DATA SYNTHESIS: Data on the associations between stage of cervical disease and response to treatment at varying levels of CD4+ lymphocyte depletion were incomplete. Recommendations by official bodies for cervicovaginal screening in women with HIV infection differed little from recommendations for standard care of all women of reproductive age. CONCLUSIONS: The consequences of a missed or delayed diagnosis of cervical disease for women with HIV infection can be severe. Pending further research, more frequent cervicovaginal screening through Papanicolaou testing and colposcopy in women with HIV infection is warranted.

Adult↗
PubMed1995

[Glycolipid analysis by confocal laser scanning microscopic system].

The combined use of immunofluorescence method with oligosaccharide-specific monoclonal antibodies against glycolipids and a confocal laser scanning microscopic system was successful to detect the storage of glycolipids in cultured cells derived from patients with lysosomal diseases. The accumulated glycolipids were observed stereoscopically as granular inclusions in the cells. An immunofluorometric semiquantative determination was achieved of globotriaosylceramide in cultured fibroblasts from Fabry disease patients or GM2-ganglioside in cultured amniocytes from Tay-Sachs disease. Heterozygote identification of the former disease or prenatal diagnosis of the latter one were confirmed by counting immunoreactive cells or by digital imaging analysis. The present immunofluorescence method is applicable to the diagnosis of the other lipidosis with accumulation of the specific glycolipids.

Animals↗
PubMed1998

[Model investigations of the impedance effectiveness conerning bacterial relevant to food hygiene].

The impedance technique mostly meets today's requirements of microbiological rapid methods. At relatively high prime cost for the equipment the advantages are marked by low personnel and material costs as well as swiftness combined with highly flexible usage. The method is applicable for both quantitative and qualitative examinations but can fail occasionally in total count determination, especially if the sample material contains heterogeneous microbes. In model investigations with 53 strains of 17 different genera Enterobacteriaceae strains, Aeromonads and Enterococcus strains proved to be highly impedance effective. Lactobacillus strains and Pseudomonads as well as Staphylococcus aureus strains showed a low impedance effectiveness. Several strains, for example of the genera Micrococcus, Acinetobacter and Brochothrix, did not show any changes of the medium impedance under the chosen conditions. Criterion for characterization of impedance effectiveness was the impedance detection time starting with identical initial counts (10(3) cfu/ml). Impedance effectiveness of microbes was determined at highly varying degree by the parameters of generation time, lag-phase duration and relative activity. This can lead either to wrong negative (underestimations) or wrong positive (overestimations) results of bacterial count.

Bacteria↗
PubMed1986

Applicability of leukocyte esterase test strip in detection of canine pyuria.

A commercially available leukocyte esterase assay was evaluated for application in analyzing canine urine for the detection of pyuria. In 229 urine samples, the leukocyte esterase activity was compared with leukocyte concentrations, as assessed by microscopic sediment analysis and chamber cell counts. The leukocyte esterase assay was specific (93.2%) for canine pyuria, but was poorly sensitive (46.0%) and did not appear to be applicable to analysis of canine urine samples.

Animals↗
PubMedDate not supplied

Automated differential cell counting of viable cells.

A rapid, automated method of differentially counting viable cells by ethidium bromide-acridine orange cytofluorography is described (Takasugi 1971). The method is shown to have comparability, equal precision, and sensitivity to the standard trypan blue, visual method. The method is applicable to microcytotoxicity testing.

Automation↗
PubMed2006

[Influence of vital dyes on the function of the outer blood-retinal barrier in vitro].

BACKGROUND: The use of vital dyes in "chromovitrectomy" allows the easier removal of less recognizable structures like epiretinal membranes (EM) or the inner limiting membrane (ILM). In recent years numerous studies demonstrated the use of indocyanine green (ICG), trypan blue (TB) and patent blue (PB) for this indication. Reports of the possible risk of these dyes, i. e. especially ICG, in respect of reduced visual acuity results, possible visual field defects or alterations of the retinal pigment epithelium (RPE) resulted in limitations in their application. MATERIAL AND METHODS: Human RPE cells and choroidal endothelial cells were cultured in monolayers on semipermeable membranes representing an in vitro model of the outer blood-retinal barrier. By measurement of the transepithelial electrical resistance (TER) the stable barrier function was determined. Two different models representing an air-filled and a fluid-filled eye were tested on the one hand by addition of the dye to the culture medium and, on the other, by direct application on the cell monolayer. In these two models ICG (5 mg/ml, 0.5 mg/ml, 0.125 mg/ml), TB (1.5 mg/ml, 0.15 mg/ml) and PB (2.4 mg/ml, 0.24 mg/ml) were applied for three minutes and the influence on the barrier function was determined. RPE cell growth was also tested in these two models after the application of ICG, TB and PB. Finally, monolayers of RPE cells were evaluated by transmission electron microscopy (TEM). RESULTS: After application of TB, PB and the lowest concentration of ICG of 0.125 mg/ml, the TER remained stable in both models. In contrast, ICG in a concentration of 5 mg/ml and 0.5 mg/ml caused a significant TER decrease in the model of the air-filled eye, whereas no influence on the function of the outer blood-retinal barrier was noted in the model of the fluid-filled eye. RPE cell growth rates were not influenced by the addition of the vital dyes, with the exception of ICG in a concentration of 5 mg/ml in the model of the air-filled eye, resulting in a temporary reduction of the cell count. In good correspondence to these results also in TEM intercellular blisters were noted after application of 5 mg/mL ICG for 3 minutes in the model of the air-filled eye. However, damage to the RPE cells themselves was not obvious. No pathological changes in the TEM were noted after application of TB and PB. CONCLUSIONS: The use of PB and TB at the posterior eye segment seems to be safe concerning damage to the PRE and its barrier function. In contrast, ICG in higher concentrations and with longer application times may cause a toxic effect on RPE morphology and function.

Blood-Retinal Barrier↗
PubMed2003

Automated enumeration of groups of marine picoplankton after fluorescence in situ hybridization.

We describe here an automated system for the counting of multiple samples of double-stained microbial cells on sections of membrane filters. The application integrates an epifluorescence microscope equipped with motorized z-axis drive, shutters, and filter wheels with a scanning stage, a digital camera, and image analysis software. The relative abundances of specific microbial taxa are quantified in samples of marine picoplankton, as detected by fluorescence in situ hybridization (FISH) and catalyzed reporter deposition. Pairs of microscopic images are automatically acquired from numerous positions at two wavelengths, and microbial cells with both general DNA and FISH staining are counted after object edge detection and signal-to-background ratio thresholding. Microscopic fields that are inappropriate for cell counting are automatically excluded prior to measurements. Two nested walk paths guide the device across a series of triangular preparations until a user-defined number of total cells has been analyzed per sample. A backup autofocusing routine at incident light allows automated refocusing between individual samples and can reestablish the focal plane after fatal focusing errors at epifluorescence illumination. The system was calibrated to produce relative abundances of FISH-stained cells in North Sea samples that were comparable to results obtained by manual evaluation. Up to 28 preparations could be analyzed within 4 h without operator interference. The device was subsequently applied for the counting of different microbial populations in incubation series of North Sea waters. Automated digital microscopy greatly facilitates the processing of numerous FISH-stained samples and might thus open new perspectives for bacterioplankton population ecology.

Automation↗