Immunohistochemical Staining of B7-H1 (PD-L1) on Paraffin-embedded Slides of Pancreatic Adenocarcinoma Tissue

B7-H1/PD-L1, a member of the B7 family of immune-regulatory cell-surface proteins, plays an important role in the negative regulation of cell-mediated immune responses through its interaction with its receptor, programmed death-1 (PD-1) ^1,2. Overexpression of B7-H1 by tumor cells has been noted in a number of human cancers, including melanoma, glioblastoma, and carcinomas of the lung, breast, colon, ovary, and renal cells, and has been shown to impair anti-tumor T-cell immunity^3-8.Recently, B7-H1 expression by pancreatic adenocarcinoma tissues has been identified as a potential prognostic marker^9,10. Additionally, blockade of B7-H1 in a mouse model of pancreatic cancer has been shown to produce an anti-tumor response¹¹. These data suggest the importance of B7-H1 as a potential therapeutic target. Anti-B7-H1 blockade antibodies are therefore being tested in clinical trials for multiple human solid tumors including melanoma and cancers of lung, colon, kidney, stomach and pancreas¹².In order to eventually be able to identify the patients who will benefit from B7-H1 targeting therapies, it is critical to investigate the correlation between expression and localization of B7-H1 and patient response to treatment with B7-H1 blockade antibodies. Examining the expression of B7-H1 in human pancreatic adenocarcinoma tissues through immunohistochemistry will give a better understanding of how this co-inhibitory signaling molecule contributes to the suppression of antitumor immunity in the tumor''s microenvironment. The anti-B7-H1 monoclonal antibody (clone 5H1) developed by Chen and coworkers has been shown to produce reliable staining results in cryosections of multiple types of human neoplastic tissues^4,8, but staining on paraffin-embedded slides had been a challenge until recently^13-18. We have developed the B7-H1 staining protocol for paraffin-embedded slides of pancreatic adenocarcinoma tissues. The B7-H1 staining protocol described here produces consistent membranous and cytoplasmic staining of B7-H1 with little background.     

Eye reduction and phylogenetic age in ophidiiform cave fish1

The West Indian Ophidiiform fish are characterized by the occurrence of several troglobitic species. They are Ogilbia (=Typhliasina pearsei (Yucatan), Lucifuga subterruneus, L. dentatus and L. simile (Cuba), and L. spelaeotes (Babama Islands). The surface relatives, e.g. Ogilbia spp., prefer lightpoor zones. This is shown by a large lens and a retina almost exclusively consistin of rods. L. dentatus, L. subterraneus, and O. pearsei possess extremely reduced eyes. The eye of L. selaeotes has morphologically an intermediate position. In correlation to varying eye size a different number of rods may be developed. Furthermore in some specimens the lens starts degenerating. Contrar to the extremely reduced cave forms all specimens of L. spelaeotes can still perceive light with tleir eyes. The different degrees of reduction in the West Indian Ophidiiform cave fish probably reflect varying phylogenetic ages. A possible date of origin of L. spelaeotes could be at the beginning or the end of the last glaciation. The eyes of neonate individuals of the live-bearing L. dentatus and O. pearsei possess a lens and a retina rudiment. No fundamental structural and histological differences between both species were found at this stage.     

Effect of concentration-time parameters on sister-chromatid exchanges induced in rabbit lymphocytes by ethylene oxide inhalation

To evaluate the effect of exposure pattern on induction and persistence of SCEs in peripheral lymphocytes and formation and persistence of the specific adduct N-3'-(2-hydroxyethyl)-histidine in hemoglobin, groups of male New Zealand white rabbits were exposed to ethylene oxide (ETO) at 200 ppm or 400 ppm for 6 h a day, 5 days a week or to 1500 ppm twice a day for 15 min until all groups reached an equal concentration-time (Ct) product of 4.8 X 10(4) ppm.h. Results show that both induced SCEs and the specific histidine adduct in hemoglobin reflect cumulative ETO exposure whether it occurs chronically at a concentration of 200 ppm or to brief exposures at the 7.5 times higher concentration of 1500 ppm. Haber's rule appears to be in effect over this range of exposure concentrations and times. Persistence of these effects appears not to be related to exposure concentration nor exposure pattern. These results contribute to further understanding of alkylating chemical mutagen dosimetry and of SCE and hemoglobin adducts as indices of exposure.     

Insertion of an intermediate repetitive sequence into a sea urchin histone-gene spacer

Summary A common polymorphism of the early embryonic histone-gene repeat ofStrongylocentrotus purpuratus is a 195-bp insertion within the H4-H2B spacer. The sequence, found as an insert in histone-gene repeats of 6 of 22 individuals screened, is also found at approximately 50 sites elsewhere in the genome of every individual. We compare the sequences of the histone-gene spacers that do and do not contain the insert. The insert is found not to have transposon-like features, and no sequence in the original spacer has been duplicated to flank the insert. There is, however, a hexanucleotide sequence that is repeated three times at one end of the insert, and the element has inserted between direct repeats of 5 bp that were present in the original spacer. One of the copies found outside the histone gene cluster was cloned and sequenced and is compared with the insert. Again, no transposon-like features are evident. Regions flanking the homologous sequence in this clone were used as hybridization probes in whole-genome blots. Results indicate that the 195-bp sequence insert is itself embedded within a larger element that is repeated within the genome. Therefore, only a portion of a larger repetitive sequence has integrated into the histone-gene spacer. The sequence features of the insert, although not typical of mobile elements, may be representative of other illegitimate recombination events.     

Skin closure with dye-enhanced laser welding and fibrinogen

The topical application of wavelength-specific dye and fibrinogen has been used to enhance laser closure of vascular anastomoses. We compared the closure of skin incisions by two different dye-enhanced, fibrinogen-based laser welding systems [argon laser (power density 4.78 W/cm2) with fluorescein isothiocyanate dye (n = 32) and diode laser (power density 9.55 W/cm2) with indocyanine green dye (n = 32)] with closure by interrupted 5-0 nylon suture (n = 64) and examined tensile strength, hydroxyproline production, histology, and cosmesis. Two 3-cm full-thickness incisions were made on the shaved backs of 64 rats. One incision was closed with suture, whereas the other, after treatment with the appropriate dye, was welded with either argon- or diode-lasered fibrinogen. At postoperative days 5, 10, 15, and 28, the closure sites were harvested and sectioned for analysis. Initially, wounds closed with argon-lasered fibrinogen showed less inflammatory response, greater collagen production (34.61 +/- 0.74 mg/gm), and greater mean peak stress at rupture (64.85 lbs/in2) than those closed with suture (16.42 +/- 3.20 mg/gm, 26.68 lbs/in2) (p less than 0.05). By 15 days, both argon and diode laser closures are superior in strength and collagen production to suture closure (p less than 0.05). At 28 days, diode laser closures (1315.60 lbs/in2) are stronger than suture closures (998.09 lbs/in2), whereas both are stronger than argon laser closures (813.16 lbs/in2) (p less than 0.05). Cosmetically, argon-welded wounds consistently appeared finer and lacked cross-hatched suture scars.(ABSTRACT TRUNCATED AT 250 WORDS)     

Physicians' attitudes toward the legalization of marijuana use.

We asked 303 practicing physicians in general internal medicine, family medicine, gastroenterology, or psychiatry to indicate whether possessing or using marijuana should be considered a felony, a misdemeanor, warrant the issuance of a citation, or be legalized. The position physicians advocated was unrelated to their specialty, experience diagnosing or treating substance abuse problems, their attitudes toward the efficacy of the treatment of drug abuse, or any other work role or habit we measured. Legalization or citation as compared with harsher penalties, however, was more likely favored by physicians who were younger, less religious, politically more liberal, and those less likely to perceive a serious drug problem in society. Legalization was also more likely favored by physicians who themselves had used marijuana, cocaine, and amphetamines but was unrelated to the use of alcohol, cigarettes, or tranquilizers. Although physician opinion should be sought as society deals with the drug problem, this study suggests how physicians'' characteristics may influence the opinions that are rendered.