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Pattern of toxicity by titanocene dichloride in mice: hematologic parameters.

Peripheral blood parameters (hematocrit, counts of RBC, reticulocytes, WBC and platelets, differential blood count) were analyzed after single injections of the cytostatic agents titanocene dichloride (TDC; ED90, 40 mg/kg; LD10, 60 mg/kg) and cisplatin (DDP; ED90, 10 mg/kg). Whereas DDP depressed the number of reticulocytes and polychromatophilic erythrocytes to near - zero level, TDC did not obviously affect the supply of young erythrocytes from bone marrow; RBC count and hematocrit values were never reduced after application either of TDC or of DDP. In the case of leukocytes, WBC count did not leave the normal range after treatment with TDC, whereas a diminution under control values was observable after application of DDP. Finally, the number of circulating platelets transiently decreased beneath control range as well after application of TDC as of DDP. The results indicate an only slight myelotoxicity induced by TDC, the platelets being the only cells affected in a reversible manner. Thus, myelosuppression by TDC is apparently even less pronounced than in the case of DDP.

Animals↗

A biphasic radiometric assay of glycogenin using the hydrophobic acceptor n-dodecyl-beta-D-maltoside.

Glycogenin is a self-glycosylating protein that catalyzes early glucosyl transfer steps in the biosynthesis of glycogen. In currently used assays of glycogenin activity, the enzyme is incubated with radioactive UDP-glucose, and the labeled reaction product is then isolated by precipitation with trichloroacetic acid. A new assay is reported here which is based on the observation that glycogenin is not only self-glycosylating but may also use exogenous alkyl maltosides as substrates. After incubation of the enzyme with n-dodecyl-beta-D-maltoside and UDP-[3H]glucose, the radioactivity in the resultant n-dodecyl-beta-D-[3H]maltotrioside is determined by any one of the following three procedures, which all rely on the hydrophobic properties conferred on the reaction product by the alkyl aglycone: (i) adsorption of the product to a Sep-Pak C18 cartridge and elution with 70% ethanol; (ii) biphasic liquid scintillation counting in ScintiLene/25% isoamyl alcohol, without isolation of the product, and (iii) precipitation with trichloroacetic acid in the presence of carrier protein. The Sep-Pak C18 procedure has the advantage that it allows essentially quantitative isolation of the reaction product, while, under the conditions chosen, only about 50% of the product is precipitated by trichloroacetic acid. For most applications, however, biphasic liquid scintillation counting is the method of choice, since close to 90% of the labeled product is extracted into the organic phase and can be counted directly without interference from the labeled nucleotide sugar which remains in the aqueous phase.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Normal blood cell values in the early mid-trimester fetus.

Samples of pure fetal blood from 116 fetuses of 15-21 weeks' gestation were obtained by direct vision fetoscopy. Ninety nine of these fetuses, presumed to be haematologically normal, were suitable for analysis. The data obtained show that the erythropoietic system is evolving rapidly in this gestational age range. The myeloid series shows no significant increase or decrease in numbers apart from eosinophils and basophils which increase significantly with gestational age whereas the platelet count remains constant. The growing application of fetoscopic blood sampling to the prenatal diagnosis and management of fetal blood disorders renders mandatory a knowledge of normal fetal blood values.

Erythrocyte Count↗

[Effect of chemical quenching on background counting rate in tritium channel of a large volume liquid scintillation counter].

Background counting rate in tritium channel of a large volume liquid scintillation counter increased with water content ranged 0 to 50% of liquid scintillator. This phenomenon can be explained as follows: The height of scintillation pulses of Compton electrons induced by background gamma radiation is lowered by chemical quenching and shifts to tritium channel. The background counting rate in tritium channel showed a linear relationship with external standard channel ratio of background samples. This relationship is applicable to determine the correct background counting rate for quenching samples and to achieve higher precision of tritium measurement.

Background Radiation↗

Application of the World Health Organization system for HIV infection in a cohort of homosexual men in developing a prognostically meaningful staging system.

OBJECTIVE: Validation of a modified version of the recently proposed World Health Organization (WHO) staging system for HIV infection and disease in a cohort of homosexual men. METHODS: Five hundred and thirty HIV-positive men followed for a median of 51 months (range, 1-98 months) were eligible for analysis. Subjects were classified into stages at their first seropositive visit and at all subsequent visits. RESULTS: As of 1 April 1991, 136 subjects (26%) had progressed to stage IV of the modified WHO system on the basis of their CD4 lymphocyte counts, and 78 subjects (15%) had died. Kaplan-Meier estimates for progression to stage IV from stages I, II and III were 52.8 +/- 7.5% over 6.6 years, 58.1 +/- 7.1% over 5.9 years and 66.5 +/- 9.7% over 5.7 years (log-rank P = 0.0001). Estimated median times to stage IV were 6.4, 5.3 and 3.8 years from stages I, II and III, respectively. Estimated median times to death were 10.9, 8.2, 6.3 and 1.7 years from stages I to IV, respectively. Results remained unchanged when CD4 lymphocyte count was replaced with lymphocyte count in the laboratory axis of the staging system. CONCLUSIONS: The proposed staging scheme, based on the WHO system, provides a prognostically meaningful classification for HIV infection in a cohort of homosexual men. Furthermore, the use of absolute lymphocyte count as a valid alternative for CD4 lymphocyte count has implications for the applicability of this system in many parts of the world where diagnostic resources are limited.

CD4-Positive T-Lymphocytes↗

Adsorption filtration of leukocyte-reduced single donor platelet units.

The effect of adsorption filtration of leukocyte-reduced single donor platelet (SDP) units collected on a CS-3000+ with a TNX chamber was compared to filtration of SDP units prepared via an isoradial chamber. The leukocyte enumeration technique employed a Nageotte chamber and had a nominal lower detection limit of 0.05 leukocytes/microliter (if no cells were seen, the concentration was recorded as 0.01 microliter). Although the prefiltration leukocyte content of the TNX units was unexpectedly low (2.4 +/- 2.8 x 10(4) per unit), filtration reduced the leukocyte content further to no greater than 1.7 +/- 0.07 x 10(3) (p < 0.05). (The full extent of the leukocyte removal could not be assessed as no leukocytes were seen in counting any of the 20 filtered TNX units). The 20 isoradial (control) SDP units achieved similar leukocyte depletion, with a postfiltration leukocyte content of 2.7 +/- 2.5 x 10(3) (p > 0.05, compared to filtered TNX units). Thus while significant leukocyte reduction occurred with adsorption filtration of both control and leukocyte-depleted SDP units, comparison of the relative degree of reduction awaits application of more sensitive leukocyte counting techniques. This study documented, however, that additional leukocyte reduction can be obtained by applying adsorption filtration to SDP units with an already low leukocyte content.

Adsorption↗

Validation of computer-assisted, pixel-based analysis of multiple-colour immunofluorescence histology.

Developments in immunohistology allow the routine simultaneous use on tissue sections of three monoclonal antibodies, tagged with different fluorochromes. Such staining can identify seven different cell populations and the limiting factor is rapid, reliable and reproducible analysis. Future reliance on computer-assisted analysis of digitised images depends on validation against manual counting, often viewed as the 'gold standard'. In this study images were digitised from sections of normal porcine skin, inflamed skin and tonsil, simultaneously stained with three monoclonal antibodies. Combinations of staining were quantified by four manual counts and by pixel-based area measurement. On individual images, the correlation between automated and manual measurements was poor. Despite this, the concordance between manual and automated measurements in the means and variances of tissues was good, and both techniques identified the same changes in inflamed versus normal tissues. In addition, pixel-based counting permitted statistical analysis of co-localisation of cell types in tissue sections. We conclude that automated counting is acceptable for the assessment of tissues, is faster and provides less opportunity for observer variation than manual counting. We also demonstrate that the technique is applicable where more than three fluorochromes are used such that manual counting becomes essentially impossible.

Animals↗

Effects of adrenaline-tablets on rat heart cells in tissue culture at normal and raised temperature.

Primary cultures of rat heart cells were incubated at 37 degrees C or 39.5 degrees C. For permanent treatment with adrenaline recently developed tablets were added to the cultures. After 10 hours the cell counts, glucose, lactate, LDH, alpha-HBDH and GOT were determined. Permanent adrenaline-application led to a decrease of cell counts, an increase of lactate, LDH, alpha-HBDH and GOT. The results indicate an injured membrane function of rat heart cells. Raised temperature sensitized the cells for adrenaline-treatment.

Animals↗

Determination of 3H-labelled cytosine arabinoside and eight metabolites in cell extracts by high-performance liquid chromatography and scintillation spectrometry.

We have developed a method for the separation and quantitation of radiolabeled cytosine arabinoside and its eight metabolites in cell extracts by anion-exchange gradient high-performance liquid chromatography. Baseline separation of cytosine arabinoside and uracil arabinoside and their respective 5'-mono-, di- and triphosphates, as well as cytosine arabinoside diphosphocholine was obtained with the shortest interval between peaks being 3 min. This degree of separation was found to be essential for quantitation of 3H-labeled metabolites by scintillation counting of 1-min fractions. Application of this procedure to the quantitation of [3H] cytosine arabinoside and its metabolites from HL60 human leukemia cells is demonstrated.

Arabinofuranosyluracil↗

A randomized clinical study of tea catechin inhalation effects on methicillin-resistant Staphylococcus aureus in disabled elderly patients.

OBJECTIVES: To evaluate the effects of tea catechin inhalation on methicillin-resistant Staphylococcus aureus (MRSA) in disabled elderly patients. DESIGN: Seven days, randomized, prospective study. SETTING: Three hospitals in Japan. PARTICIPANTS: Seventy-two patients aged 78 +/- 11 years (mean age +/- standard deviation) with cerebrovascular diseases, classified as disabled according to the activity of daily living and were either bedridden or required assistance for standing, and showing presence of MRSA in sputum. INTERVENTIONS: Inhalation of 2 mL tea catechin extract solution along with saline (3.7 mg/mL catechins, 43% of catechins are composed of epigallocatechin gallate), or saline alone, 3 times daily using a handheld nebulizer for 7 days. MEASUREMENTS: The endpoint of efficacy was the reduction rates of MRSA in sputum. The safety measure was the adverse events observed during the 7 days of inhalation. RESULTS: The reduction rates calculated as the summation of decrease and disappearance of MRSA in sputum at 7 days were 47% (17 of 36 patients) in the catechin group and 15% (5 of 33 patients) in the control group; the difference in the reduction rates between the 2 groups was statistically significant (P = .014). The disappearance rate of MRSA in sputum was higher in the catechin group (31%; 11 patients) when compared with the control group (12%; 4 patients), however the difference in the disappearance rate between the 2 groups was not statistically significant (P = .091). No adverse events, such as respiratory tract obstruction, allergic bronchial spasm, or skin eruption, including laboratory changes, were observed during the study. CONCLUSION: The catechin inhalation appeared to reduce the MRSA count in sputum. However, the application of tea catechin inhalation as a supplementary treatment for controlling MRSA infection remains controversial.

Administration, Inhalation↗

Ampicillin killing curve patterns for ampicillin-susceptible nontypeable Haemophilus influenzae strains by the agar dilution plate count method.

Ampicillin killing curve patterns for 20 strains of ampicillin-susceptible nontypeable Haemophilus influenzae were determined by the agar dilution plate count method. The paradoxical effect was detected in the 24-h killing curve patterns for each strain. For the biphasic effect, minimum survivor percentages (maximum killing) occurred over a narrow range of ampicillin concentrations immediately above the MIC, with survivor percentages then rising rapidly to peak at approximately 1-log10-unit increment higher. The 24-h minimum survivor percentages for the 20 strains ranged from approximately 0.01% (rapid killing) to greater than 10% (slow killing). In comparison with the previous results for typeable strains, the present findings suggest that nontypeable stains are, on average, killed much more slowly. Based on the initial 24-h killing curve patterns for the 20 strains, 4 strains were selected as putative representatives of the range of bactericidal responses encountered. These strains were then studied to examine the reproducibility of the 24-h patterns and to determine sequential killing curves. These patterns were found to be reproducible and served to characterize the relative killing responses of the strains. In the sequential studies of three of the four strains, tiny colonies having the gross and microscopic characteristics of L-forms were found to be present on the agar dilution plate count plates prior to the application of penicillinase at 48 and 72 h. Such colonies reverted to vegetative forms within 24 to 48 h after application of penicillinase to the panels. Of particular interest was the observation that the paradoxical effect was manifested both by the L-form colonies and by the reverted vegetative colonies. The late development of L-forms was observed for both rapidly and slowly killed strains.

Ampicillin↗

Photon counting computed tomography: concept and initial results.

A concept of a photon counting cone beam CT is proposed. The system uses a new Multi Slit Multi Slice (MSMS) cone beam acquisition geometry utilizing a linear array photon counting detectors. The MSMS cone beam acquisition is a direct analogy of the scanning multislit acquisition used in projection x-ray imaging. This geometry provides a CT imaging with dose efficient scatter rejection and allows for using available photon counting detectors. The microchannel plate (MCP) detector is proposed as a linear array photon counting detector for MSMS cone beam CT system. Initial testing of the MCP detector for CT application was performed. The field of view of the prototype MCP detector is 60 mm. A delay line position encoding electronics was used. The electronics has a single channel input for evaluation of events from the entire detector field of view. This limits the system count rate at 2 x 10(5) count/s. The spatial resolution of this detector is 80 microm FWHM at 40 kVp and 200 microm FWHM at 90 kVp tube voltages. The detector noise in CT projections is less than 1 count/pixel for the 80 microm pixel size. The CT projections contain quantum-limited and scatter free signal. Images of a contrast phantom and a small animal were acquired at 50 kVp and 80 kVp tube voltages. The CT numbers for different contrast elements were calculated for a given x-ray spectrum and compared with experimental values. The quantum efficiency of the current detector is 56% at 90 kVp, which is suboptimal because of the large channel diameter (25 microm) of these MCPs. The MCPs with smaller channels and higher efficiencies are being tested. The quantum efficiency was measured to be 70% for a new MCP with 5 microm channel diameter. Design parameters of a clinically applicable photon counting MSMS cone beam CT for breast imaging was evaluated. System uses 20 cm field of view MCP detectors based on 5 microm channel MCPs and high count rate ASIC electronics. It was concluded that the MSMS cone beam CT with a photon counting MCP detector is feasible for volume breast imaging.

Breast Neoplasms↗

Role of nitric oxide synthase inhibition in leukocyte-endothelium interaction in the rat pial microvasculature.

We investigated the role of nitric oxide (NO) in leukocyte-endothelium interaction, blood-brain barrier (BBB) function and oxygen free-radical production in the rat pial microcirculation. In a closed cranial window preparation (dura removed) over the parietal cortex of pentobarbital-anesthetized Wistar rats, NO synthase (NOS) was inhibited by systemic and/or topical application of N omega-nitro-L-arginine (L-NNA) under physiological conditions and during leukotriene B4 (LTB4) activation. Circulating leukocytes were labeled by intravenous injection of rhodamine 6G. We used a confocal laser scanning microscope (CLSM) and studied leukocyte rolling and sticking in pial veins and arteries before and after NOS inhibition. At the end of the experiments, sodium-fluorescein was injected intravenously to test BBB integrity. Brain cortex oxygen free-radical production was investigated in the cranial window preparation using lucigenin-enhanced chemiluminescence (CL). L-NNA application did not lead to significant changes in leukocyte-endothelium interaction, BBB function, and oxygen free-radical production under physiological conditions [leukocyte-endothelium interaction: control (n = 5), L-NNA systemically (n = 5), L-NNA topically (n = 5): at baseline rollers/100 microns: 0.76 +/- 0.55, 0.64 +/- 0.94, 0.44 +/- 0.55 and stickers/100 microns: 0.90 +/- 0.28, 0.76 +/- 0.24, 0.84 +/- 0.42; at 60 min rollers/100 microns: 1.49 +/- 0.66, 1.21 +/- 0.99, 0.67 +/- 0.66 and stickers/100 microns: 1.04 +/- 0.20, 1.19 +/- 0.23, 1.21 +/- 0.54; oxygen free-radical production (n = 4): CL count before L-NNA application 35 +/- 17 cps, after 1 h of topical superfusion of L-NNA 38 +/- 14 cps; p < 0.05]. In contrast to the results achieved under physiological conditions, a significant further increase of rolling leukocytes and BBB permeability occurred due to NOS inhibition under LTB4-activated conditions [76 +/- 47% significant (p < or = 0.01, n = 7) further increase of rollers/100 microns due to 60 min L-NNA application following the activation period of 120 min LTB4 superfusion]. Our results support a modulatory role for NO in leukocyte-endothelium interaction and BBB permeability in the pial microcirculation when this interaction is increased.

Animals↗

The effect of method of infection on the pathway of juvenile Strongyloides ratti in the host.

Subcutaneous injection of the larvae is the almost universally adopted means of initiating experimental infections of skin-invading roundworms but, so far, the possibility that this procedure introduces artefacts of one kind or another has not been critically studied. Experiments described in this paper were used to compare the effect of (a) injection and (b) skin application, of a small, precisely counted ('exact') dose of larvae. Results with two strains of S. ratti showed that the same proportion of the dose developed to adults in the intestines of rats irrespective of the method. With the same exact dose technique it has been shown that milk-borne infection of the pups of lactating rats is not an artefact produced by injection. Large doses (mean 4000) of larvae of the homogonic strain of S. ratti carrying a radioactive label of 75Se were tracked in their migration to the mammary gland following injection or skin application at two different sites on the right-hand side of nursing mother rats. The broad conclusion of earlier work in this laboratory using injection, that larvae move by a local route and not a systemic one, was supported by the results. The detailed distribution of the label and of unlabelled worms of the heterogonic strain in families was, however, different for the two methods, indicating that subtle variations in pathway can be brought about by the use of injection. If migration involves the lymphatic system, then the interpretation of immunological experiments in terms of lymphatic anatomy must take account of such procedural effects. The extent to which these results contribute to theories of migration in Strongyloides ratti is discussed.

Anesthesia↗

Number of dishes as an indicator of nutrient intake in China.

The objectives of this analysis are to investigate if counting the number of dishes consumed per day is a rapid procedure for estimating daily nutrient intake in China, and to explore if urban-rural differences exist in the daily number of dishes. A nutritional survey (including dish number counting) was conducted on 499 adult women in six urban and four rural communities in China. The number of dishes was an influential variable in estimating intake of protein, animal protein, fat, animal fat, and some vitamins (e.g., vitamin B2) and minerals (e.g., zinc), but not of energy and carbohydrate. Intake of some nutrients (e.g., protein) was sufficient when people consume more than 10 dishes per day. Thus, the number of dishes is a useful indicator of sufficient intake of animal protein and fat as well as some micro-nutrients. The application of the dish number counting method showed that there was a significant difference between urban (11.9 dishes per day) and rural populations (7.9 dishes).

Adult↗

Use of Healon in different cataract surgery techniques: endothelial cell count study.

The effect of the intracameral application of 1% sodium hyaluronate (Healon) in various cataract extraction techniques was evaluated in four groups of ten cats each whose natural lenses were removed using four different techniques. In a fifth group, the intraocular lenses (IOLs) were implanted over the crystalline lens. Each animal's right eye was treated with Healon and each left eye with balanced salt solution (BSS). Healon-treated eyes lost almost 50% fewer endothelial cells and had a smaller increase in corneal thickness than their counterpart eyes treated with BSS. Lens implantation and phacoemulsification procedures caused the greatest endothelial cell loss, while neodymium YAG laser capsulotomy, a less traumatic procedure, minimized this loss.

Animals↗

Application of simultaneous determination of 3H, 14C, and 22Na by liquid scintillation counting to the measurement of cellular ion-transport.

A liquid scintillation counting method for simultaneous determination of three radioactive nuclides (3H, 14C, and 22Na) of biological interest was studied. By comparing the beta spectra of the three nuclides, their counting energy ranges, A, B, and C, were determined. 22NA was set high enough to avoid any spillover counts from lower-energy nuclides. Region A for 3H was set to maximize the counting efficiency. A good correlation between the counting efficiency for 22Na in region C and the counting efficiency of other nuclides in all regions was obtained. Prior to 3H and 14C dpm calculations, the 22Na counts spilled down in regions A and B were subtracted from the total counts in regions A and B. A simple linear equation was then used to compute 3H and 14C dpm. Findings show that the method presented is adaptable for highly quenched samples up to quenching indices of tSIE = 100. The method is useful for studying the biological transport coupled to Na+.

Beta Particles↗