MORPHOMETRICAL PARAMETERS OF GUT MICROFLORA IN HUMAN VOLUNTEERS

The morphology of faecal microflora of nine healthy human volunteers was studied by digital image analysis of microscopic slides. Weekly specimens were collected during an 8-week period. Seven morphometrical parameters were derived: the means and medians of components 1, 2 and 3, and morphometrical entropy. Statistically significant differences among subjects were found for means of components 1 and 2. medians of components 1, 2 and 3, and entropy. The stability in normal circumstances provides an excellent basis for the detection of pathological change in gut flora balance

Gaussian-weighted moving-window robust automatic threshold selection

A multi-scale, moving-window method for local thresholding based on Robust Automatic Threshold Selection (RATS) is developed. Using a model for the noise response of the optimal edge detector in this context, the reliability of thresholds computed at different scales is determined. The threshold computed at the smallest scale at which the reliability is suffcient is used. The performance on 2-D images is evaluated on synthetic an natural images in the presence of varying background and noise. Results show the method deals better with these problems than earlier versions of RATS at most noise levels

Effects of ceftriaxone on faecal flora:analysis by micromorphometry

In order to elucidate the effect of ceftriaxone therapy on the morphology of gut microflora, 11 human volunteers were treated with ceftriaxone, 1 g daily, given intramuscularly in one dose. Treatment continued for 5 days. Faecal microflora was analysed by digital image processing before, during and after the treatment period. We derived simple numerical parameters which describe the morphologic composition of the flora. They were significantly influenced by the antibiotic, and returned to their baseline values more than 7 days after treatment was stopped. The procedure holds promise for clinical application

Immunocytochemical analysis of cisplatin-induced platinum-DNA adducts with double-fluorescence video microscopy.

To detect low-level DNA platination, a sensitive immunocyto- and histochemical technique was developed using a polyclonal antibody. The antibody GPt, derived after immunization of rabbits with highly platinated DNA and purified with affinity chromatography, detected the main platinum (Pt)-containing intrastrand and interstrand adducts. Double-fluorescence microscopy image analysis was used to quantify Pt-DNA adducts with Hoechst 33258 fluorescence to locate the nuclei and with fluorescein isothiocyanate fluorescence to measure the immunosignal. A two- to five-fold dose-dependent difference in the level of cisplatin (CDDP)-induced Pt-DNA adducts between a CDDP-sensitive and -resistant human tumour cell line was detected. Large differences in Pt-DNA adduct levels after in vitro CDDP incubation between human buccal cells, lymphocytes and biopsies of different tumour types were observed. Pt-DNA adduct levels were fivefold higher in human testicular tumours than in colon tumours, representing CDDP-sensitive and -resistant tumours, respectively, in the clinic. These data suggest the possibility of predictive testing by measuring Pt-DNA adduct levels. Pt-DNA adducts in patients after treatment with CDDP were shown in normal buccal cells and in imprints of fresh tumour biopsies as well as in paraffin-embedded tumour cells. The analysis of Pt-DNA adducts at a single-cell level in small samples of normal and tumour cells during and/or after treatment is feasible with GPt and will hopefully enable more selective treatment of patients