PubMed · 14458093
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A KOMAN, L CZABAFY. 1961. [Experience with an automatic cell counter].. https://pubmed.ncbi.nlm.nih.gov/14458093/
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As part of a learning exercise to assess the efficiency of oocyte utilization after controlled ovarian stimulation, we compared historical data from the publications of Steptoe and Edwards describing their early experiences in Oldham and at Bourn Hall with retrospective data from IVF and ICSI cycles carried out at Bourn Hall Clinic during the year 2000. For the purpose of analysing the more recent data, patients were subdivided into groups by age (< or = 37 years and > or = 38 years) and by oocyte yield (low, intermediate and high). In patients less than 38 years of age, live birth rates were equivalent in each group, irrespective of the number of oocytes retrieved. Patients who yielded low numbers of oocytes (1-5) utilized an average of 9.6 oocytes per live birth, compared with 25.1 and 51.5 in those who yielded intermediate (6-16) and high (16+) numbers of oocytes. The comparison with historical data suggests that the 'efficiency' of oocyte utilization has not improved significantly since the early 1980s, despite 25 years' experience of controlled ovarian stimulation for IVF treatment.
Flow cytometric signatures (i.e., light scatter, red and green fluorescence) were obtained for the active but non-culturable (ABNC) cells of E. coli and a coliform isolate H03N1, in seawater microcosms using BacLight, a live-dead assay kit from Molecular Probes (Eugene/Portland, OR). Previous studies have reported that there are two major adaptations, which cells undergo during the formation of ABNC states: cell wall toughening and DNA condensation. Therefore, we hypothesized that the matured ABNC forms should be more resistant to extreme temperature treatments (i.e., by freezing in liquid nitrogen and thawing at room temperature) than the normal and transition populations. It was shown that the membrane-compromised cells (comprising of normal wild-type and dead cells which are less resistant to rapid freeze thaw) could be differentiated from the matured ABNC using BacLight staining and fluorescence detection by flow cytometry. The population of ABNC cells, which could not be cultured using m-FC media (for the enumeration of fecal coliforms), was resuscitated in phosphate buffer saline followed by growth in Luria broth. Flow cytometry was thus able to detect and differentiate the ABNC cells against a mixed population comprising of culturable cells, transition populations, and dead cells. The results also showed that the formation of ABNC is as early as 2 days in seawater microcosms. By directly comparing the coliform levels enumerated by the BacLight based flow cytometry assays and m-FC technique, it was shown that the presence of coliforms can be undetected by the membrane filtration method.
OBJECTIVES: To characterize and compare the histological features of conjunctiva obtained during the repair of trabeculectomy and inadvertent blebs and to assess the effects of antifibrotic agents. METHODS: We used routine paraffin processing and light microscopy to examine conjunctival specimens from 28 eyes undergoing late bleb revision: 19 leaking trabeculectomy blebs, 5 nonleaking trabeculectomy blebs, and 4 inadvertent blebs. Quantitative analysis of histological characteristics included conjunctival epithelial thickness, stromal vascularity, squamous metaplasia, stromal inflammatory cell infiltrate, and goblet cell density. The type of surgery, presence of leaks, prior topical medication, and antifibrotic use were correlated with histological findings. RESULTS: Trabeculectomy blebs (leaking and nonleaking) showed decreases in overall epithelial thickness (P<.02), goblet cell density (P<.001), and vascularity (P<.001) compared with inadvertent bleb controls. Conjunctiva exposed to antifibrotics had fewer layers of epithelial cells than those with no prior exposure (P<.05). CONCLUSIONS: Histological analysis of thin blebs excised following trabeculectomy demonstrated decreases in epithelial thickness and goblet cell density compared with inadvertent blebs. Both leaking and nonleaking trabeculectomy blebs exhibited decreases in localized stromal vascularity and increases in surrounding stromal vascularity compared with normal conjunctiva.