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PubMed1989

The implications of the heterogeneous distribution of Ki-67 labelled cells in meningiomas.

Ki-67 monoclonal antibody reacts with a nuclear protein only expressed in proliferating cells. In order to evaluate the applicability of Ki-67 in meningiomas we determined the index of labelled cells in 52 biopsies. Two counting methods were used to calculate the labelling index. The mean Ki-67 index was determined by the number of positive Ki-67 cells in three randomly chosen fields divided by the total number of cells in the three fields. The preparations were also screened for the area with the highest percentage of positive Ki-67 cells, yielding the 'highest' Ki-index. The three patients who developed a recurrence did not show an elevated Ki-67 index by any of the two counting methods. Because of the differences observed between the two counting methods and the differences of the Ki-67 indices in the three randomly chosen fields we performed a statistical analysis to evaluate the possibility of a heterogeneous distribution of Ki-67 positive cells in meningiomas. Homogeneity was rejected if the data showed no Poisson distribution. We found that five of the 13 tumours which had a mean Ki-67 index greater than 1.00% were heterogeneous (38.4%). One of the highest differences between the two counting methods (5.05%) was noted in the only tumour with local signs of malignancy. Our results indicate a heterogeneity of the labelling of Ki-67 in histologically benign meningiomas and emphasise the importance of representative sampling. More attention should be paid to this heterogeneity for cell kinetic studies in brain tumours, and one should be aware that regrowth is not only determined by the proliferative capacity.

Antibodies, Monoclonal↗

PubMed2001

In vivo evaluation of amitraz against ticks under field conditions in Ethiopia.

An aqueous emulsion of amitraz (Bovitraz, Bayer AH), prepared and applied according to the manufacturer's recommendations, was evaluated for its activity against cattle ticks on a dairy farm in Ethiopia. Eight crossbred heifer calves aged between 6 and 8 months and heavily infested with ticks were selected and divided into equal treatment and control groups. The calves in the treatment group were hand-sprayed with the amitraz emulsion while the control group was left untreated. Each calf was restrained and ticks were counted and identified in situ. One hundred percent tick control was achieved on Day 3 after acaricide application, and this was maintained for a further 18 days. The residual effect was long, and protected the animals from re-infestation for 21 days. There was a significant difference between the mean number of ticks on the control group compared to the treated group (P < 0.05). An important finding in this trial was the rapid detachment of all tick instars from animals treated with the test acaricide.

Animals↗

PubMed2002

Responses of songbirds to aerial spraying of the microbial insecticide Bacillus thuringiensis var. kurstaki (FORAY 48B) on Vancouver Island, British Columbia, Canada.

Use of bacterial insecticides containing Bacillus thuringiensis var. kurstaki (Btk) is gaining popularity as an environmentally safe control measure against lepidopteran pests, but indirect effects on nontarget organisms through reduced prey base have received little attention. Aerial spraying of Btk (Foray 48B) over a 12,803-ha area on southeastern Vancouver Island (BC, Canada) in May to June 1999 as part of a gypsy moth (Lymantria dispar) control program provided us with an opportunity to examine the responses of songbirds to spray application. To obtain an estimate of species richness and relative abundance, we conducted standard songbird point-count surveys in Garry oak (Quercus garryana)-dominated habitats in Btk-sprayed and unsprayed areas in April to June 1999 and, one year after spraying, in 2000. These surveys revealed no patterns consistent with adverse effects of spraying on the relative abundance of adult birds or singing males for any of the species, whether examined individually or when combined into foraging guilds. An exception was the spotted towhee (Pipilo maculatus), which in 1999, but not in 2000, occurred at significantly lower numbers in sprayed plots after Btk treatment. Intensive searches of plots in sprayed and unsprayed areas revealed no differences in the numbers of songbird broods between the two areas for any of the species examined.

Animals↗

PubMed1991

Correlation between tissue growth kinetics and modulation of mouse skin tumorigenesis by phorbol esters.

Previous studies on the influence of phorbol esters on mouse skin tumorigenesis have shown that 12-O-tetradecanoylphorbol-13-acetate (TPA) enhances development of malignant epithelial and mesenchymal skin tumors by a completely carcinogenic dose of 3-methylcholanthrene (MCA), while its congener phorbol-12, 13-diacetate (PDA) exerts an inhibitory effect. Differential effects of these two agents were analysed by histology, morphometry and cell kinetic techniques including autoradiography and estimation of labelled precursor incorporation into DNA by liquid scintillation counting. Epidermal hyperplasia induced on exposure of S/RV Cri mouse skin to a single or multiple TPA application after MCA injection was associated with a significant increase in the thickness of nucleated cell layers, stratum granulosum, number of suprabasal cells and dark basal cells. Enhancing effect of TPA on MCA-induced neoplastic development correlated well with an increase in mitotic activity, number of cells in S-phase and increased rate of DNA synthesis in the epidermis, dermis and subcutis as also mast cell number. In contrast, treatment of MCA-injected preneoplastic mouse skin with PDA resulted in epidermal hypoplasia and cellular damage evident as cytoplasmic vacuolation and nuclear pyknosis. Multiple PDA exposure also reduced the thickness, mitotic index and number of cells in S-phase in epidermis, dermis and subcutis. Thus, cellular toxicity and inability to recruit cells in DNA-synthetic phase may account for inhibition of progression of preneoplastic epithelial and mesenchymal cells into overt tumors by PDA.

Animals↗

PubMed1986

The epidermal Langerhans' cell population in psoriasis during topical coal tar therapy.

We have studied the effect on the Langerhans' cell (LC) population of topical 3% coal tar therapy. Biopsies were taken from psoriatic plaques and from controls with no skin disease before and after the application of 3% coal tar for one week; LC were identified by immunofluorescence using monoclonal antibody. LC counts expressed per unit epidermal surface length were similar in untreated psoriasis plaques and in normal skin. Differences in the LC population in paired biopsies from both patients and controls showed considerable variation following coal tar treatment but no consistent effect could be demonstrated.

Antibodies, Monoclonal↗

PubMed1989

[Initial experience with autologous thrombocytes labeled with 111indium-oxine].

The authors elaborated their own modification of the separation and labelling of autologous thrombocytes by means of 111indium-oxine which can be implemented in Czechoslovak departments of nuclear medicine with standard equipment. A total of 123 separations and labellings were made and 85 administrations to patients. The effectiveness of labelling by the described method was 79.5 +/- 6.9%, the separation yield of thrombocytes with regard to complete blood 44.3 +/- 13.8%. The aggregating capacity of the labelled platelets assessed in vitro after ADP stimulation (adenosine diphosphate) was 39.5 +/- 10.1% difference in optic density of platelet rich and platelet poor plasma which is evidence of the preserved viability of thrombocytes. "Recovery" during the 120th minute after administration of normal suspensions was 62.2 +/- 17.9%, the survival time of labelled thrombocytes 8.2 +/- 0.74 days. In patients with idiopathic thrombocytopenic purpura the "recovery" was 49.4 +/- 18%, the survival period 2.9 +/- 1.8 days. In patients after transplantation of the kidney complicated by acute rejection there was an increased accumulation of labelled platelets in the area of the kidney (count rate index 2.32 +/- 0.45). In normally functioning transplanted kidneys the count rate index was 1.42 +/- 0.06. On their own material the authors demonstrate other possibilities of clinical application of scintigraphy by means of labelled autologous platelets (a method hitherto not described in the Ccezhoslovak literature) for the detection of intracardial and arterial thrombosis.

Blood Platelets↗

PubMed1988

Reproducibility of mitotic counts and identification of mitotic figures in malignant glial tumors.

Quantitative pathology is a rapidly growing field of pathology. This field is developing methods allowing greater objectivity and better reproducibility in diagnostic decisions. The introduction of quantitative pathology as a teaching subject in medical education will help this field to become more widely and more correctly applied. Courses on quantitative pathology are organized yearly by the Department of Pathology at the University of Ancona. The 1986 course emphasized training of students in understanding basic problems such as variation caused by tissue processing, instrumentation, data handling and interpretation, and variation between observers. In this paper we describe experiments performed during the course on identification and quantitation of mitoses in malignant glial tumors. The participants, i.e. students of the postgraduate Pathology School, could identify mitoses with a 'substantial' reproducibility. However, when the observers were allowed freely to choose the fields to count, the reproducibility was 'slight'. The study suggests that sampling rules should be applied which secure reproducible application of morphometric method also on condition of free selection of fields. Because the application of such rules needs expertise in histopathology, it seems that morphometric method can best be applied by histopathologists themselves.

Glioma↗

PubMed1987

Regional brain hematocrit in stroke by single photon emission computed tomography imaging.

Nineteen studies on 18 subjects were performed by single photon emission computed tomography (SPECT) of the head after the successive intravenous administration of a plasma label (99mTc-human serum albumin [HSA]) and 99mTc-labeled autologous red blood cells (RBC). Two sets of cerebral tomographic sections were generated: for cerebral 99mTc-HSA alone and for combined 99mTc-HSA and 99mTc-RBC. By relating counts in regions of interest from the cerebral tomograms to counts from blood samples obtained during each tomographic acquisition, regional cerebral haematocrit (Hct) was calculated by the application of a simple formula. Results show 1) lower cerebral Hct than venous Hct (ratio of HCT brain/Hct venous 0.65-0.90) in all subjects, and 2) comparison between right and left hemisphere Hct in 3/3 normal subjects, 6/6 patients with transient ischaemic attacks and 3/8 patients with stroke showed no significant difference. However, in 3/8 patients with stroke (most recent strokes) significant differences were found, the higher Hct value corresponding to the affected side.

Adult↗

PubMed1993

Fast chemiluminescent measurement of RNA polymerase activity based on photon counting technology.

A fast and simple assay for T7 RNA polymerase based upon chemiluminescent detection of the synthesized, digoxigenin-labeled RNA on a nylon membrane with anti-digoxigenin coupled alkaline phosphatase and CSPD as substrate is described. Activity of RNA polymerase is determined with high sensitivity by quantifying the emitted light of the microplate-formatted dot-blot membrane with a photon counting microplate luminometer and a specially designed filter adapter. The described method is one example for the application of this new adapter to measure luminescent membrane filters.

DNA-Directed RNA Polymerases↗

PubMed1993

[Heparin-associated thrombocytopenia as a problem in heparin therapy].

The clinical picture of HAT (heparin-associated thrombocytopenia) is being described in reference to a typical case report. If not recognized in time, HAT may lead to serious thrombo-embolic complications, which may eventually lead to death. Considering the wide applications of routine heparin in therapy and prophylaxis, HAT deservers more clinical attention. Routine monitoring of platelet counts during heparin treatment is crucial and essential.

Aged↗

PubMed1976

Whole blood microculture assay of human lymphocyte function.

A whole blood microculture assay is described for measuring lymphocyte reactivity to mitogenic and antigenic stimulants. This assay employs heparinized whole blood, serum-free culture medium, microtiter plates, and a Multiple Automated Sample Harvester (MASH). When this assay is compared to other leukocyte assays, its major advantages include (1) the utilization of fewer lymphocytes per microculture, thuus reducing the amount of blood required per test while increasing the number of test agents and replicate cultures which can be employed in any given experiment; (2) the conservation of mitogens, antigens, drugs, enzymes, hormones, lymphokines, and other test agents, some of which are either expensive of difficult to prepare in large quantities; (3) the elimination of lymphocyte isolation and purification procedures which may disrupt the relative proportion of T cells, B cells and antigen-processing cells; and (4) the application of an automated harvester which simplifies and expedites procedures required for processing cells for liquid scintillation counting.

Cell Separation↗

PubMed1995

Plaque bacteria counts and vitality during chlorhexidine, meridol and listerine mouthrinses.

The aim of this double-blind study was to enumerate the total number of living and dead bacteria on defined tooth areas during the application of antibacterial mouthrinses. After prophylaxis, 40 students refrained from all oral hygiene measures for 3 d, during which they rinsed with a phenolic compound (Listerine), an amine fluoride/stannous fluoride solution (Meridol), 0.2% chlorhexidine (CHX) or a control solution (0.02% quinine-hydrochloride). The plaque index (P1I) was recorded at the start and the end of the investigation. Total bacterial counts (BC) and colony-forming units (CFU) of 1d-, 2d- and 3d-old dentogingival plaque were determined. The plating efficiency (PE) was calculated as a percentage of CFU/BC and the portion of vital microflora estimated by a vital fluorescence technique (VF). All groups started with a P1I approximating 0.1. On day 3, the P1I values were 1.21 in the control group and 0.51, 0.37 and 0.14 after Listerine, Meridol and CHX use, respectively. A tremendous variation existed between the numbers of viable bacteria found per mm2 on the enamel surface and day 3 (CHX: 0.2; Meridol: 300; Listerine; 6x10(4); control: 2x10(6)), while higher total numbers of bacteria were concomitantly present (CHX and Meridol: 1-2x10(4); Listerine: 2x10(5); control: 2x10(6)). Both vitality parameters PE and VF reached 92% in the control group at day 3, but only 7% after CHX use. With Meridol and Listerine, the corresponding PE values were 3% and 43%, respectively, while the VF values reached 48% and 54%. The PII, BC, CFU and PE values of the CHX and the Meridol groups differed significantly from those of the control group. In contrast, Listerine showed no difference as compared to the control rinse. Due to the strong antibacterial action of CHX and Meridol during their use, almost only dead or non-proliferating bacteria were found on the tooth surfaces. Thus, only a thin plaque could develop. As a clinical consequence, both substances showed retardation of plaque development as reflected by significantly reduced plaque indices.

Adolescent↗

PubMed2002

A retrospective study of the effect of long-term topical application of retinaldehyde (0.05%) on the development of actinic keratosis.

BACKGROUND: The role of topical retinoids on photocarcinogenesis is still unclear. Retinaldehyde is a natural metabolite of vitamin A used as a cosmetic product. Its effect on actinic keratoses has not been studied to date. OBJECTIVE: To study the incidence of actinic keratoses during long-term application of retinaldehyde in order to evaluate a possible chemoprophylactic effect. METHODS: We conducted a retrospective study on 61 patients who had applied retinaldehyde on photoexposed body areas for a period ranging from 6 to 142 months. We counted the total number of actinic keratoses and cutaneous tumors that appeared over the time of exposure to retinaldehyde. RESULTS: The epidemiological characteristics of actinic keratoses were not modified by the application of retinaldehyde. Irregular application as compared to regular application of retinaldehyde was not associated with a change in the risk of actinic keratoses, suggesting that continuous use is not associated per se with a higher risk of actinic keratoses. CONCLUSION: With the statistical power limitation of this study, retinaldehyde applied alone does not appear to have prophylactic effects on the development of actinic keratoses. The design adopted is feasible to study the safety of cosmetic products applied for a long period of time.

Administration, Topical↗

PubMed1989

Evaluation of PET count rate performance.

A proposal is made for the test conditions to evaluate PET count rate performance. This performance depends in a complex manner on the spatial distribution of activity and scattering material. Therefore, a combined body phantom is proposed, which is as simple as possible but which adequately simulates the range of clinical application of a whole body tomograph. Taking into account the special properties of the new block detector design, a comprehensive test procedure is developed. This includes not only the common count rate characteristic, but also checks for the accuracy of randoms estimation and count loss correction schemes, and for the occurrence of pulse pile up. This is done for different source and scatter configurations, simulating brain, cardiac, and abdominal imaging, respectively. Examples are given, based on measurements of the latest generation PET scanners, namely the CT1 PT 931/08-12 and the Scanditronix PC 2048-07WB.

Models, Structural↗

PubMed1999

Depletion of alveolar macrophages by treatment with 2-chloroadenosine aerosol.

Alveolar macrophages (AMs) are localized in the alveoli and alveolar ducts of the lung and are the only macrophages living in an aerobic environment. Recent studies have demonstrated that AMs play a central role in lung diseases, such as pneumonia and acute respiratory distress syndrome. It has become important to find a simple, effective way to eliminate AMs in order to investigate the function of AMs in vivo. 2-Chloroadenosine (2-CA), a purine analog, is reported to be selectively cytotoxic to cultured macrophages, and we hypothesized that it would deplete the number of AMs in the bronchoalveolar lavage fluid (BALF) of mice without any effect on neutrophil or lymphocyte counts. After mice had inhaled 1 mM aerosolized 2-CA for 2 h, AMs were found to be significantly depleted at 0 h [(4.42 +/- 0.16) x 10(4)/ml], 24 h [(4.17 +/- 0.89) x 10(4)/ml], 48 h [(3.17 +/- 0.21) x 10(4)/ml], and 72 h [(5.00 +/- 0.64) x 10(4)/ml] compared with concentrations in untreated controls [(12.1 +/- 0.21) x 10(4)/ml]. Neutrophil and lymphocyte counts in BALF did not change and histological changes in the lung were not observed after 2-CA treatment. The lung wet-to-dry weight ratio did not change at 0, 24, and 48 h after 2-CA aerosol application. The 2-CA aerosol had no effect on lung vascular permeability, as assessed by the intravenous administration of Evans blue, or on other phagocytes, as assessed by Kupffer cell counts. Our study demonstrates the efficacy of 2-CA in reducing AM numbers in vivo.

2-Chloroadenosine↗

PubMed1999

Activity of a deltamethrin shampoo against Ctenocephalides felis and Rhipicephalus sanguineus in dogs.

A controlled clinical trial was undertaken to assess the efficacy of an application of a 0.07% deltamethrin shampoo against fleas and ticks in dogs. Twenty beagles, housed in separate cages, were randomly allocated into two groups of 10. The dogs were infested with 50 Ctenocephalides felis and 50 Rhipicephalus sanguineus each, and parasites were counted 24 h (fleas) and 72 h (ticks) later (Day 0). Dogs, when state, were then treated with 25 ml of a shampoo containing 0.07% deltamethrin (Group 1). Dogs in Group 2 were the controls. Fleas and ticks were counted 24 h (Day 1) and 48 h (Day 2) after the shampoo was applied, and the parasites were then removed. All the dogs were reinfested with fleas and ticks on Days 2, 7, 9, 14, 16 and 20. Parasites were counted 24 and 48 h after each reinfestation. Effectiveness against fleas was calculated 24 h after infestation, and against ticks 48 h after infestation. This study showed that the application of 25 ml of shampoo containing 0.07% deltamethrin to beagle dogs weighing between 10.2 and 12 kg was very well tolerated. It controlled the parasites present on the animals at the time of application: with an efficacy of 100% against fleas 24 h after treatment and an efficacy of 95% against ticks 48 h after treatment. The treatment protected against flea reinfestations with an efficacy of 100% during the first week, >98% in the second week and >95% in the third week. It also gave >99% protection against tick reinfestations in the first week and >96% in the second week.

Animals↗

PubMed2003

Pneumocystis carinii pneumonia as a complication of bendamustine monotherapy in a patient with advanced progressive breast cancer.

BACKGROUND: Bendamustine is an alkylator with anticipated antimetabolic activity. It has shown activity in malignant lymphoma, multiple myeloma, and breast cancer. Recognized side-effects are relatively mild with myelosuppression as the dose-limiting toxicity. The CD4/CD8 ratio may be reduced. To what extent the alteration of lymphocytes, especially CD4(+) lymphocytes, correlates with an increase in opportunistic infections cannot be definitively answered. CASE REPORT: The patient, female, aged 48 years, was suffering from an advanced progressive breast cancer. After initial treatment with several chemotherapies, a cytotoxic therapy was initiated, with bendamustine (150 mg/m(2)) administered on two consecutive days and repeated every 4 weeks. After five courses, the patient developed Pneumocystis carinii pneumonia (PCP), disclosed in the bronchoalveolar lavage. While receiving bendamustine therapy, the CD4(+) and CD8(+) lymphocyte counts in the peripheral blood were determined by flow cytometry. The next-to-normal CD4/CD8 ratio before therapy (0,82) had decreased to 0,05 during the therapy mainly due to a decline of CD4(+) lymphocyte. The patient was seronegative for human immunodeficiency virus. In spite of high-dose intravenous trimethoprim/sulfamethoxazole and methylprednisolone application, the patient died of a respiratory failure 3 days after PCP was diagnosed. CONCLUSION: Bendamustine is capable of inducing a reduction in CD4(+) lymphocyte counts causing a severe T-lymphocyte-mediated immunosuppression. Measuring CD4(+) lymphocyte counts may be helpful in determining the risk of PCP in patients treated with bendamustine.

Antineoplastic Agents↗

PubMed2004

Quantitative PCR with 16S rRNA-gene-targeted species-specific primers for analysis of human intestinal bifidobacteria.

A highly sensitive quantitative PCR detection method has been developed and applied to the distribution analysis of human intestinal bifidobacteria by combining real-time PCR with Bifidobacterium genus- and species-specific primers. Real-time PCR detection of serially diluted DNA extracted from cultured bifidobacteria was linear for cell counts ranging from 10(6) to 10 cells per PCR assay. It was also found that the method was applicable to the detection of Bifidobacterium in feces when it was present at concentrations of >10(6) cells per g of feces. Concerning the distribution of Bifidobacterium species in intestinal flora, the Bifidobacterium adolescentis group, the Bifidobacterium catenulatum group, and Bifidobacterium longum were found to be the three predominant species by examination of DNA extracted from the feces of 46 healthy adults. We also examined changes in the population and composition of Bifidobacterium species in human intestinal flora of six healthy adults over an 8-month period. The results showed that the composition of bifidobacterial flora was basically stable throughout the test period.

Adult↗