Autofluorescence imaging can identify preinvasive or clinically occult lesions in fallopian tube epithelium: a promising step towards screening and early detection.

BACKGROUND: Optical imaging systems are robust, portable, relatively inexpensive, and have proven utility in detecting precancerous lesions in the lung, esophagus, colon, oral cavity and cervix. We describe the use of light-induced endogenous fluorescence (autofluorescence) in identifying preinvasive and occult carcinomas in ex vivo samples of human fallopian tube (FT) epithelium. METHODS: Women undergoing surgery for an i) ovarian mass, ii) a history suggestive of hereditary breast-ovarian cancer, or iii) known serous ovarian cancer following neoadjuvant chemotherapy (NAC) were approached for informed consent. Immediately following surgery, FT's were photographed in reflectance and fluorescence at high resolution. Images included: (1) white-light reflectance of luminal/epithelial surface; (2) narrow-band green reflectance (570 nm) (3) green autofluorescence (405/436 nm excitation); and (4) blue autofluorescence (405 nm excitation). Areas revealing a loss of natural tissue fluorescence or marked increase in tissue microvasculature were recorded and compared to final histopathologic diagnosis (SEE-FIM protocol). RESULTS: Fifty-six cases involving one or both fallopian tubes underwent reflectance and fluorescence visualization. Nine cases were excluded, either secondary to non-ovarian primary pathology (7) or excessive trauma (2) rendering tissue interpretation impossible. Of the 47 cases remaining, there were 11 high grade serous (HGS) and 9 non-serous ovarian carcinomas undergoing primary debulking surgery, 5 serous carcinomas having received NAC, 8 benign ovarian tumors, and 14 women undergoing risk-reducing bilateral salpingo-oophorectomy (RRBSO). Methodology was feasible, efficient, and reproducible. TIC or carcinoma was identified in 7/11 HGS, 3/5 NAC, and 1/14 RRBSO. Optical images were reviewed to determine test positive or negative based on standardized criteria. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated for the entire cohort (73%; 83%; 57%; 91%) and in a subgroup that excluded non-serous histology (87.5%; 92%; 78%; 96%). CONCLUSIONS: Abnormal FT lesions can be identified using ex vivo optical imaging technologies. With this platform, we will move towards genomic interrogation of identified lesions, and developing in vivo screening modalities via falloposcopy.

Pyrene is metabolized to bound residues by Penicillium janthinellum SFU403.

We have previously shown that the filamentous fungus, Penicillium janthinellum SFU403 (SFU403) oxidizes pyrene to pyrene 1,6- and 1,8-quinones and that the level of pyrenequinones (PQs) subsequently declines suggesting that PQs are not terminal metabolites. The purpose of this study was to determine the fate of PQs in SFU403. First, we compared the fate of 14C-pyrene in SFU403 and a non-pyrene-oxidizing fungus, a Paecilomyces sp.. After 7 days of incubation, more than 80% of the radioactivity was cell-associated in both fungi; however, while 90% of the 14C could be extracted from the Paecilomyces sp. as unmetabolized pyrene, 65-80% of the bound radioactivity remained inextractable from SFU403. Further evidence that pyrene oxidation to PQs was required for irreversible binding was obtained by comparing the extent of 14C bound to SFU403 when it was grown for 21 days under conditions that resulted in differing amounts of 14C-pyrene oxidation. The results showed that approximately 40% of the inextractable products were bound residues derived from pyrene metabolites. The balance (60%) could be attributed to strong sorption of unreacted pyrene. We used electron paramagnetic resonance spectroscopy and oxygen consumption studies to demonstrate that both NADPH and glutathione can reduce PQs by one electron to their corresponding semiquinone anion radicals in vitro. These studies demonstrate that PQs are metabolized by SFU403 to bound residues, possibly via semiquinone intermediates.

Radioimmunoassay of salivary cyclosporine with use of 125I-labeled cyclosporine.

We prepared 125I-labeled cyclosporine (125I-CS) by modifying the procedure of Mahoney and Orf (Clin Chem 1985;31:459) and characterized it with regards to maximal immunoreactivity (greater than 90%), trichloroacetic acid precipitability (greater than 90%), and stability (90% immunoreactive after five half-lives of 125I). For a particular preparation of 125I-CS, we estimated its immunoreaction concentration (50 pmol/L) and the equilibrium constant for its reaction with Sandoz polyclonal antiserum (K = 3.9 X 10(9) L/mol). By substituting 125I-CS as tracer in the Sandoz radioimmunoassay and by modifying other aspects of the assay, we developed a procedure that is sufficiently sensitive (0.34 micrograms/L) to allow measurement of "trough" (lowest inter-dose) cyclosporine concentrations in parotid saliva. Of 38 kidney-transplant patients, 35 had measurable concentrations in saliva (mean 8.3, SD 5.2 micrograms/L), and these correlated moderately with paired serum concentrations (r = 0.68, P less than 0.001). We believe that measurement of salivary cyclosporine may offer a simple way of estimating the free fraction of the drug in serum or plasma.

Long-term follow-up of children with femoral neck fractures.

Of 92 children reviewed three to five years after treatment for fractured neck of femur (Lam 1976), we have been able to reassess 41, both clinically and radiographically, at 13 to 23 years after injury. The earlier clinical results had been excellent, despite a high incidence of complications; but the new, later review shows an 83% incidence of radiographic abnormality while 24% of the patients have pain, a limp or leg shortening. We present a recommended policy for management of this rare but potentially serious childhood injury.