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Potential efficacy of a delta 5-aminolevulinic acid thermosetting gel formulation for use in photodynamic therapy of lesions of the gastrointestinal tract.

Photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA)-induced protoporphyrin IX may play a role in the treatment of dysplastic Barrett's oesophagus. An ALA thermosetting gel Pluronic F-127) was developed and evaluated in an in vivo mouse model for potential use in PDT of Barrett's mucosa. In vitro studies of the influence of Pluronic F-127 percentage on thermosetting gel temperature, followed by the influence of ALA concentration on thermosetting temperature and ALA-gel stability as a function of time or temperature were studied. In vivo relationships between ALA doses and fluorescence were studied to determine the optimal concentration. Fluorescence measurement in vivo showed that ALA concentration and time had a nonlinear influence on protoporphyrin IX synthesis. For ALA-gel applications longer than 30 min a plateau fluorescence was reached, the maximum fluorescence being obtained after 4 h whatever the time of contact. The maximum intensity (2824 counts s(-1)) was found with 40 mg mL(-1) ALA-gel, and fluorescence intensities differed with time, reaching a maximum after 3-4 h. ALA-Pluronic F-127 is a suitable formulation for treatment of Barrett's oesophagus, allowing easy application in liquid form at 4 degrees C and good adhesion in the oesophagus in gel form, with efficient diffusion of ALA into treated mucosa.

Aminolevulinic Acid↗

Effect of liposome-entrapped methotrexate on Ehrlich ascites tumor cells and uptake in primary liver cell tumor.

A therapeutically useful concentration (0,5 mg/ml) of Methotrexate was prepared in negatively charged artificial liposomes (lecithine: cholesterol: dicetylphosphate=5:5:1 molar ratios). Entrapment yield after separation on Sepharose 6B is nearly 100%. In contrast to free Methotrexate the liposome entrapped folic acid antagonist is eliminated only slowly by the kidneys and after intravenous application it is unevenly distributed in the body of mice, the highest concentrations being found in liver and spleen. Daily injections (7.5 mg/kg/day) of entrapped Methotrexate for 5 days into the tail vein of Ehrlich ascites tumor bearing mice reduced both tumor cell count and the production of ascites fluid about fourfold as compared to mice receiving the same dose of Methotrexate in the free form. Six hours after intravenous application of liposome entrapped Methotrexate the tissue concentration in normal liver is ca. 20-fold higher than when the same amount is applied in the free state. On the other hand, only a little uptake of liposome entrapped Methotrexate was detected in the tissue of nitrosamine-induced primary liver tumor when compared to normal liver tissue of rats.

Animals↗

Diagnostic laparoscopy in the horse.

Laparoscopy was performed in 5 clinically normal horses. Abdominal fluid protein content and WBC count increased markedly in all horses. Necropsy findings were confined to an increased amount of slightly turbid abdominal fluid. Five examples of applications of laparoscopy in horses with abdominal problems indicated that laparoscopy can be a valuable diagnostic procedure in the horse.

Abdomen↗

Application of a finite mixture model to somatic cell scores of Italian goats.

The objectives of this study were to apply a finite mixture model (FMM) to data for somatic cell count in goats and to compare the fit of the FMM with that of a standard linear mixed effects model. Bacteriological information was used to assess the ability of the model to classify records from healthy or infected goats. Data were 4518 observations of somatic cell score (SCS) and bacterial infection from both udder halves of 310 goats from 5 herds in Northern Italy. The records were from a complete production season, and were taken monthly from February to November 2000. Explanatory factors in both models included a 3-parameter regression on days in milk (DIM); fixed class effects of herd-test-day, parity group, and udder side (left or right); and random effects of goat and udder half within goat. In addition, the 2-component FMM included a fixed mean for the second component of the model (theoretically corresponding to infected udder halves), as well as an unknown probability of membership to a given putative infection status. A Bayesian statistical approach was used for the analysis with Gibbs sampling used to obtain draws from posterior distributions of parameters of interest. Two sampling chains of 200,000 cycles each were generated for each model. The FMM yielded a much lower estimate of residual variance than the standard model (1.28 vs. 3.02 SCS2), and a slightly higher estimate for the between-goat variance (1.79 vs. 1.48). The deviance information criterion (DIC) was used to compare the fit of the 2 models. The DIC was much lower for the FMM, indicating a better fit to the data. The FMM was able to classify correctly 60 and 48% of the healthy and infected observations, respectively. This was slightly higher than what would be expected from random classification, but not high enough for useful mastitis diagnosis. Nevertheless, increased precision of genetic evaluation is the goal of applying the FMM, rather than timely and accurate mastitis diagnosis. The results suggest that more research on FMM for SCS is merited and necessary for proper application.

Analysis of Variance↗

Problems associated with the direct viable count procedure applied to gram-positive bacteria.

Despite the numerous advantages of fluorescent in situ hybridization (FISH) for identifying a single bacterial cell with 16S rRNA probes, problems are encountered with starving bacteria in natural samples. The original direct viable count procedure (DVC) includes a revivification step in the presence of an antibiotic inhibiting cell division. Cells elongate and accumulate ribosomes. This results in a natural amplification of 16S rRNA molecules (target of FISH). However, it is limited to gram-negative bacteria which are sensitive to nalidixic acid. The objective of this study was to develop a procedure for estimating the number of metabolically active gram-positive Staphylococcus aureus and Enterococcus faecalis cells by the use of a method which combines the number of substrate-responsive cells and their identification by FISH. It was observed that no single published DVC method could apply to taxonomically different gram-positive bacteria. Since cells were not counted, the revivification step in presence of nalidixic acid will be referred to as revivification without cell division. For each species, different low-nutrient media and complex media, different fluoroquinolones and beta-lactam antibiotics, concentrations of antibiotics, combinations of antibiotics, temperature and time were evaluated using bacteria in different physiological states and in natural samples. Enumeration of bacteria by plate counts and direct FISH were compared. The improved procedure should yield information about the physiological state, the taxonomic identity, and the enumeration of viable gram-positive bacteria. The application of DVC to an entire ecosystem is presently still a challenge.

Anti-Bacterial Agents↗

Caries-preventive effect of topical amine fluoride in children with high and low salivary levels of mutans streptococci.

The aim of the study was to assess the relationship between the salivary mutans streptococci (SMS) level and the effectiveness of a preventive intervention based on a biannual application of an amine fluoride solution (AmF). A total of 284 schoolchildren aged 6 years were recruited from eleven classes of a primary school in Milan and randomly assigned to an experimental (A) and a control group (B). SMS counts were obtained at baseline and caries incidence data (diseased, missing, filled teeth, DMFT) were recorded every 6 months for 5 years. The participants of the experimental group received application of an AmF 1% F(-) solution twice a year on the enamel surfaces of the first permanent molars for 5 years. Control group subjects received application of a placebo solution twice a year on the enamel surfaces of the first molars for the same period. The mean DMFT in the experimental and control groups were 0.56 and 0.22, respectively, at the beginning and 1.14 and 2.06 after 5 years. SMS data allowed children to be classified into low- (0-10(5) CFU/ml of saliva) and high- (>10(5) CFU/ml of saliva) SMS subjects. Survival analysis, performed on the first molar data split by SMS group, showed a significantly higher caries reduction in low-SMS experimental group subjects compared to low-SMS control group subjects after 5 years. No significant differences were found between the two high-SMS experimental and control groups. These findings indicate that the preventive effects of the treatment were significantly lower in subjects who had high SMS. It is concluded that the effectiveness of a simple and economical topical fluoride intervention applicable at a community level is significantly influenced by the SMS level of the subjects involved. Simple AmF preventive interventions, applied on low-SMS subjects, can give significant results in terms of caries reduction.

Administration, Topical↗

Specific molecular detection of Carnobacterium piscicola SF668 in cold smoked salmon.

AIMS: To establish a rapid and reliable multiplex PCR (mPCR)-based method allowing specific identification of Carnobacterium piscicola SF668 during storage of cold smoked salmon (CSS). METHODS AND RESULTS: CSS was inoculated with C. piscicola SF668 and stored at 4 degrees C. Samples were withdrawn at regular time intervals and analysed by counting the number of viable cells. About 25-100% of colonies grown on Elliker plates were subjected to mPCR amplification. The results show that strains presumably identified as C. piscicola SF668 were predominant over the test period. CONCLUSIONS: mPCR is a powerful tool to study competitiveness of C. piscicola SF668, which inhibits the growth of Listeria monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study demonstrates the importance of molecular methods in studying competitiveness of strains with potential food applications.

Animals↗

Positive end-expiratory pressure depressed cardiovascular autonomic nervous system activity in acute brain damaged rabbits under general anesthesia.

Artificial ventilation with positive end-expiratory pressure (PEEP) is commonly applied for brain damaged patients. However, the effect of the ventilation on brain function, including cardiovascular autonomic nervous system (CANS) activity, is not well elucidated. In order to investigate the effect of 5 cmH2O PEEP on CANS activity in brain damaged rabbits under general anesthesia, we produced acute brain damage by intracranial balloon inflation. Measurements were made before (control) and after application of PEEP, and after inflation with incremental volume of the balloon. Power spectral analyses of heart rate variability (HRV) and systolic arterial pressure variability (SAPV) were used for the assessment of CANS activity. Spectral powers in the low-frequency range of 0.04 to 0.40 Hz (LF) and high-frequency range of 0.75 to 1.40 Hz (HF) were computed, and their ratio LF/HF was assessed as the neural balance of CANS. The animals in group P were ventilated with 5 cmH2O PEEP, while those in group Z were ventilated with zero end-expiratory pressure. Colored microsphere counting was used for the assessment of brain circulation. In the results, PEEP had no effect on HRV and SAPV parameters before induction of brain damage. After inflation with incremental volume of the balloon, log (HF) and log (LF) in group P were lower than in group Z in HRV analysis, and log (LF) in group P was lower than in group Z in SAPV analysis. Microsphere counting revealed that brain blood flow was reduced during the progression of brain damage and showed a significant difference after application of PEEP between the groups. We concluded that 5 cmH2O PEEP depressed CANS activity during the progression of brain damage in rabbits and that this was partly due to aggravated brain function induced by PEEP.

Anesthesia, General↗

Dose-response relationships of alimentary PtCl2 and PtCl4 in growing rats.

In experiments with rats dose-response relationships of alimentary PtCl2 and PtCl4 were investigated. 2 x 81 animals weighing 35 g were randomly distributed among 9 treatment groups which were fed ad libitum with a synthetic diet containing various amounts of Pt during 4 weeks. Pt was added in the form of PtCl2 or PtCl4 in the amounts 0; 0.01; 0.05; 0.10; 0.50; 1.0; 5.0; 10 and 50 mg/kg diet. The Pt supplementation had no influence on life mass gain or food consumption. In the case of 50 mg/kg Pt in the form of PtCl4 the erythrocyte count and hematocrit were reduced by about 13% in comparison with the control group. Dependent on the Pt dose, the application of PtCl4 and PtCl2 induced Pt retention in nearly all tissues especially in kidney. The effects were greater with PtCl4 than with PtCl2. As a result of the higher Pt retention in the kidneys, the serum creatinine was increased for the higher doses of PtCl4.

Animals↗

Development of large platelet aggregates from small aggregates as determined by laser-light scattering: effects of aggregant concentration and antiplatelet medication.

Particle-counting methods that employ light scattering (LS) quantify changes in the number of platelet aggregates of different sizes after the application of an aggregating stimulus. Using the LS method, we studied the effects of aggregant concentration, aspirin administration, and ticlopidine administration on aggregate formation and compared the results with those obtained using the conventional optical density (OD) method. Subjects were 47 controls, 31 patients treated with aspirin (330 mg/day), and 37 patients treated with ticlopidine (200 mg/day). Platelet aggregation after stimulation by 0.5, 1.0 or 5.0 muM ADP, or 0.5, 1.0 or 2.0 micrograms/ml collagen was determined using both methods. Using the LS method, small (9-25 micrograms), medium (25-50 micrograms), and large (50-70 micrograms) aggregates were counted. In patients untreated with antiplatelet medication, greater concentrations of ADP or collagen generated larger aggregates. Generation of small and medium-sized aggregates showed a significant positive correlation with OD levels after stimulation with 0.5 or 1.0 muM ADP, or 0.5 or 1.0 micrograms/ml collagen. In patients treated with aspirin, the development of small aggregates into large aggregates was inhibited. Thus, the number of small aggregates increased. Inhibition induced by aspirin was more effective against aggregation after stimulation with collagen than with ADP. In patients treated with ticlopidine, small and medium-sized aggregate formation was inhibited after stimulation with low concentrations of ADP or collagen, but was promoted after stimulation with high aggregant concentrations. The capability of the LS method to quantify different sizes of aggregates after stimulation with low concentration agonists may facilitate investigation of the aggregation process, and of how this process is affected by antiplatelet agents.

Aged↗

99mTc-pyrophosphate kinetics in man (relationship between bone uptake and mineral content).

Both the kinetics and the regional distribution of 99mTc-PPi were studied in the skeleton of healthy subjects of various ages by surface counting using a scintillation counter placed over different skeletal areas. The results were compared with bone mineral content as determined by the photonic absorption technique. Kinetic data of bone uptake were obtained by subtracting the counts measured in blood from the counts recorded over each body surface after making proportional corrections. The surface-counting patterns show a higher generalized 99mTc-PPi bone uptake in children and adolescents than in adults. The relative regional distribution of the tracer shows no difference between the two groups. The tracer accumulated more rapidly in spongy bone than in compact bone, and the fixation process was carried out during the first 30 min for both spongy and compact bone. In youths, however, higher uptake values corresponded to lower values of mineral content. The findings enable the normal surface-counting pattern to be constructed and confirm that the 99mTc-PPi uptake is related to bone metabolism; they also suggest the application of external measurements for the study of metabolic bone diseases.

Adolescent↗

Hematopoietic growth factors: overview and clinical applications, Part I.

The growth and differentiation of blood cells is regulated by a group of at least 12 glycoproteins, collectively known as hematopoietic growth factors. Advances in protein biochemistry and molecular genetics have provided the tools for the bulk production of these hormones for clinical application. Clinical trials of macrophage colony-stimulating factor, granulocyte macrophage colony-stimulating factor, granulocyte colony-stimulating factor, and interleukin-3 have all demonstrated significant effects on the peripheral blood counts of the recipients. The clinical usefulness of these agents in ameliorating post-chemotherapy myelosuppression, in the treatment of other cytopenias, and in enhancing engraftment after bone marrow transplantation has already been demonstrated. Potential applications to the therapy and diagnosis of other clinical disorders is under study. The history of the elucidation of these growth factors, our current understanding of their properties, interactions, and clinical effects, and the potential prospects for their future use in the manipulation of human blood cell production are the subject of this review.

Cells, Cultured↗

A study on methods of estimating the population size of men who have sex with men in Southwest China.

OBJECTIVE: To ascertain practical methods of estimating the population size of men who have sex with men (MSM) by field application. METHODS: Nine public places were selected, where men who have sex with men (MSM) frequented. Capture-recapture method and multiplier method were used to count the number of men who have sex with men (MSM) in the assigned places in 10 days. RESULTS: The population size estimated using multiplier method counted 877. Three estimates produced by capture-recapture method were 1408 (95% CI: 1116-1908), 1207 (95% CI: 932-1712) and 949 (95% CI: 757-1272), respectively. CONCLUSIONS: Multiplier method was easy and smooth in practice and produced highly reliable results, which yet was expensive. The application of capture-recapture method cost less in both time and funds, and the three results produced could verify each other, which was fit for the estimation.

China↗

Colchicine ultrasensitivity of lymphocytes in chronic lymphocytic leukaemia.

It has been previously reported that, in comparison with normal lymphocytes, the lymphocytes in chronic lymphocytic leukaemia (CLL) are ultrasensitive in culture to the cytocidal action of colchicine. In this report the results of 240 colchicine studies in 87 patients with CLL are presented and analysed in terms of the diagnostic, clinical and haematological significance of colchicine ultrasensitivity (CUS) in CLL. All patients with CLL showed very significantly increased CUS of lymphocytes. The CUS did not vary with clinical and haematological status or time and only very rarely with treatment. There was no correlation between % CUS and subsequent course or survival. The results in CLL and in other forms of lymphocytosis and in non-Hodgkin's lymphoma are compared. It is concluded that the test is of considerable diagnostic value, particularly in identifying low-count and treated CLL and in the exclusion of non-CLL lymphocytosis. It appears to have no prognostic value but may have future application in monitoring treatment which selectively eliminates the abnormal cell population.

Adult↗

An effective hormonal male contraceptive using testosterone undecanoate with oral or injectable norethisterone preparations.

Suppression of spermatogenesis to azoospermia is the goal of hormonal male contraception based on T combined with gestagens. The combination of the long-acting T, ester testosterone undecanoate (TU), with norethisterone (NET) enanthate (E) showed high efficacy. In the present study, we tested the validity of this approach by varying the NET dose and mode of application. The aim of the study was to achieve high rates of suppression of spermatogenesis as reflected by sperm counts, monitor gonadotropins as well as other hormones, and evaluate any possible side effects. In a phase II clinical trial, groups of normal volunteers received: 1000 mg TU im at wk 2, 6, 12, and 18 combined with 200 mg NETE im at wk 0, 6, 12, and 18 (group I); 1000 mg TU im and 400 mg NETE im at wk 0, 6, 12, and 18 (group II); and 1000 mg TU im at wk 0, 6, 12, and 18 with daily oral NET acetate (NETA) from wk 0 to 24 (group III). In all groups marked suppression of gonadotropins resulted in a significant decrease of spermatogenesis and azoospermia in 13/14, 11/12, and 12/14 men in groups I to III, respectively. The remaining men all had less than 1 million sperm/ml. Reversible side effects included increase in body weight, erythrocytes, hemoglobin, and hematocrit and decrease in high-density lipoprotein cholesterol and alkaline phosphatase in all groups and increase in liver enzymes in the oral NETA group. This study documents the high efficacy of TU in combination with NET and confirms that this dose and mode of application (1000 mg TU im every 6 wk plus 400 mg NETE im every 6 wk or plus 10 mg daily oral NETA) is as effective as the previously reported regimen containing 1000 mg TU + 200 mg NETE im every 6 wk. The contraceptive efficacy of this combination of TU and NETE should be evaluated in further clinical trials.

Administration, Oral↗

Sensitivity of test for overdispersion in Poisson regression.

Overdispersion or extra-Poisson variation is very common for count data. This phenomenon arises when the variability of the counts greatly exceeds the mean under the Poisson assumption, resulting in substantial bias for the parameter estimates. To detect whether count data are overdispersed in the Poisson regression setting, various tests have been proposed and among them, the score tests derived by Dean (1992) are popular and easy to implement. However, such tests can be sensitive to anomalous or extreme observations. In this paper, diagnostic measures are proposed for assessing the sensitivity of Dean's score test for overdispersion in Poisson regression. Applications to the well-known fabric faults and Ames salmonella assay data sets illustrate the usefulness of the diagnostics in analyzing overdispersed count data.

Bias↗