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J Stahl

Publications and source records attributed to J Stahl.

At least 19 recordsLinked to original sources

Apolipoprotein-D: a novel cellular marker for HGPIN and prostate cancer.

BACKGROUND: High grade prostatic intraepithelial neoplasia (HGPIN) is a putative pre-malignant lesion of the prostate. While apolipoprotein-D (Apo-D), an androgen-regulated hydrophobic transporter protein, is expressed in prostate tumors, its expression in HGPIN is unknown. METHODS: Immunoreactivity for Apo-D and another androgen-regulated protein, prostate specific antigen (PSA), was investigated in 64 radical prostatectomy tissues by video image analysis. RESULTS: Eighty two percent of prostatectomy specimens demonstrated moderate to strong Apo-D immunoreactivity in areas of HGPIN. In comparison, weak Apo-D immunoreactivity was observed in non-malignant areas in only 24% of specimens. The median (range) percentage cellular area of HGPIN immunopositive for Apo-D (9.7%, 0-42.9), and the cellular concentration of Apo-D (MIOD 3.1, 0-13.3), were intermediate between that of normal (area 0%, 0-53.5%, MIOD 0, 0-12.6) and early stage prostate cancer tissues (area 29.2%, 0-90.8%, MIOD 6.7, 0-28.1). This increase in Apo-D expression from non-malignant, through HGPIN to prostate cancer was statistically significant (P < 0.001), and contrasted with the decrease observed in PSA staining between adjacent areas of normal glands, HGPIN, and cancer (P = 0.026). CONCLUSIONS: The presence of high levels of immunoreactive Apo-D in HGPIN and prostate cancer, but not in non-malignant epithelial cells, is consistent with HGPIN being an intermediate lesion in the transition to prostate cancer, and suggests that cellular Apo-D expression is a marker of malignant transformation of the prostate.

Apolipoproteins↗

Signal transducer gp130: biochemical characterization of the three membrane-proximal extracellular domains and evaluation of their oligomerization potential.

Glycoprotein 130 (gp130) is a type I transmembrane protein and serves as the common signal-transducing receptor subunit of the interleukin-6-type cytokines. Whereas the membrane-distal half of the gp130 extracellular part confers ligand binding and has been subject to intense investigation, the structural and functional features of its membrane-proximal half are poorly understood. On the basis of predictions of tertiary structure, the membrane-proximal part consists of three fibronectin-type-III-like domains D4, D5 and D6. Here we describe the bacterial expression of the polypeptides predicted to comprise each of these three domains. The recombinant proteins were refolded from solubilized inclusion bodies in vitro, purified to homogeneity and characterized by means of size-exclusion chromatography and CD spectroscopy. For the first time the prediction of three individual membrane-proximal protein domains for gp130 has been verified experimentally. The three domains do not show intermediate-affinity or high-affinity interactions between each other. Mapping of a neutralizing gp130 monoclonal antibody against D4 suggested a particular functional role of this domain for gp130 activation, because above that an intrinsic tendency for low-affinity oligomerization was demonstrated for D4.

Animals↗

Ribosomal protein S5 interacts with the internal ribosomal entry site of hepatitis C virus.

Translational initiation of hepatitis C virus (HCV) genome RNA occurs via its highly structured 5' noncoding region called the internal ribosome entry site (IRES). Recent studies indicate that HCV IRES and 40 S ribosomal subunit form a stable binary complex that is believed to be important for the subsequent assembly of the 48 S initiation complex. Ribosomal protein (rp) S9 has been suggested as the prime candidate protein for binding of the HCV IRES to the 40 S subunit. RpS9 has a molecular mass of approximately 25 kDa in UV cross-linking experiments. In the present study, we examined the approximately 25-kDa proteins of the 40 S ribosome that form complexes with the HCV IRES upon UV cross-linking. Immunoprecipitation with specific antibodies against two 25-kDa 40 S proteins, rpS5 and rpS9, clearly identified rpS5 as the protein bound to the IRES. Thus, our results support rpS5 as the critical element in positioning the HCV RNA on the 40 S ribosomal subunit during translation initiation.

5' Untranslated Regions↗

Growth arrest in A549 cells during hyperoxic stress is associated with decreased cyclin B1 and increased p21(Waf1/Cip1/Sdi1) levels.

Exposure to high concentrations of oxygen has previously been shown to cause growth arrest in A549 cells, a distal lung epithelial cell line. We found that when A549 cells were exposed to 95% oxygen they underwent substantial growth inhibition. This was associated with induction of p21(Waf1/Cip1/Sdi1) protein and a decrease in cyclin B1 protein. Flow cytometry revealed that A549 cells exposed to hyperoxia had a significant decrease in the percentage of cells in G(1) and a modest but significant increase in the percentage of cells in S phase and G(2)/M, consistent with cells entering S phase. A549 cells in room air and hyperoxia were then treated with nocodazole, a mitotic inhibitor. Room air A549 cells treated with nocodazole showed a marked increase in G(2)/M consistent with mitotic arrest. In contrast, hyperoxic treated cells had a modest but significant decrease in G(1) but only a minimal increase in G(2)/M consistent with partial G(1)/S arrest and growth inhibition in S phase. To further investigate the role of p21(Waf1/Cip1/Sdi1) as a checkpoint regulator during hyperoxic growth inhibition, HCT116 cells with wild-type and null p21(Waf1/Cip1/Sdi1) were exposed to hyperoxia. Both wild-type p21(+/+) cells and null p21(-/-) cells underwent growth inhibition when exposed to hyperoxia. At 48 h the hyperoxic treated HCT116 p21(+/+) had a similar cell cycle distribution as the hyperoxic treated HCT116 p21(-/-) cells, suggesting that p21(Waf1/Cip1/Sdi1) may not be essential for growth arrest during hyperoxia. These findings suggest that hyperoxia causes partial growth arrest at different phases of the cell cycle but primarily in S phase, that hyperoxic growth arrest is associated with a decrease in cyclin B1 protein and that p21 induction may not be essential for hyperoxic growth arrest.

Animals↗

Radiological-pathological correlation: alveolar pattern.

This review summarizes important pathological lesions of the lung that typically present radiographically with an 'alveolar pattern'. For each entity, the latest findings as to its pathogenesis, aetiology and pathology are reviewed in the introductory remarks. We then present the typical radiological appearances alongside macroscopic and microscopic pathological photographs. It is hoped that the parallel presentation of radiological image with the pathology will enhance the understanding of the diverse range of diseases the aevolar pattern comprises.

Diagnosis, Differential↗

Evaluation of a waterless, scrubless chlorhexidine gluconate/ethanol surgical scrub for antimicrobial efficacy.

A new waterless surgical hand preparation containing 1% chlorhexidine gluconate (CHG) and 61% ethyl alcohol was evaluated for antimicrobial efficacy in comparison with a standard 4% CHG surgical scrub and a 61% ethyl alcohol control. Clinical studies were based on the Tentative Final Monograph for Health-Care Antiseptic Drug Products (TFM) (proposed rule) and the Standard Test Method for Evaluation of Surgical Hand Scrub Formulations (ASTM E1115-91). Two randomized, blinded, well-controlled clinical studies involving 137 healthy subjects were conducted to evaluate the antimicrobial effectiveness of the CHG/ethanol hand preparation in producing an immediate and persistent reduction in the normal bacterial flora of the hands. The CHG/ethanol hand preparation was applied without scrubbing or the use of water, and a standard 4% CHG reference product was applied with a scrub brush in 2 traditional 3-minute surgical scrubs. In 1 study, a 61% ethanol vehicle control treatment was applied without scrubbing or use of water. During a 5-day period, each study subject performed a series of 11 surgical scrubs with 1 of the test treatments. After the first treatment on days 1, 2, and 5, surgical gloves were worn for 3 or 6 hours. Bacterial samples were taken with the glove-juice technique at 1 minute, 3 hours, and 6 hours after treatment. The immediate bactericidal effect of the CHG/ethanol hand preparation after a single application resulted in a 2.5-log reduction in normal flora. This bactericidal effect persisted throughout the studies and eventually increased to a 3.6-log reduction after the 11th scrub on day 5. The log reductions of the CHG/ethanol hand preparation proved to be significantly better (P <.05) than that of the 4% CHG product at each sampling interval on days 1 and 2 and the sampling at 6 hours on day 5 and significantly better than the 61% ethanol vehicle at all times. The combination of 1% CHG and 61% ethanol had significantly greater microbial reduction than either the 4% CHG (without ethanol) or the 61% ethanol vehicle (without CHG).

Adult↗

Assessment of two hand hygiene regimens for intensive care unit personnel.

OBJECTIVE: To compare skin condition and skin microbiology among intensive care unit personnel using one of two randomly assigned hand hygiene regimens: a 2% chlorhexidine gluconate (CHG)-containing traditional antiseptic wash and a waterless handrub containing 61% ethanol with emollients (ALC). DESIGN: Prospective, randomized clinical trial. SETTING: Two critical care units (medical and surgical) in a large, metropolitan academic health center in Manhattan. SUBJECTS: Fifty staff members (physicians, nurses, housekeepers, respiratory therapists) working full time in the intensive care unit. INTERVENTIONS: One of two hand hygiene regimens randomly assigned for four consecutive weeks. MEASUREMENTS AND MAIN RESULTS: The two outcomes were skin condition (measured by two tools: Hand Skin Assessment form and Visual Skin Scaling form) and skin microbiology. Samples were obtained at baseline, on day 1, and at the end of wks 2 and 4. Participants in the ALC group had significant improvements in the Hand Skin Assessment scores at wk 4 (p = 0.04) and in Visual Skin Scaling scores at wks 3 (p = 0.01) and 4 (p = 0.0005). There were no significant differences in numbers of colony-forming units between participants in the CHG or ALC group at any time period. The ALC regimen required significantly less time than the CHG regimen (mean: 12.7 secs and 21.1 secs, respectively; p = 0.000) and resulted in a 50% reduction in material costs. CONCLUSIONS: Changes in hand hygiene practices in acute care settings from the traditional antiseptic wash to use of plain, mild soap and an alcohol-based product should be considered. Further research is needed to examine the association between use of antiseptic products for hand hygiene of staff and reductions in nosocomial infection rates among patients.

Adult↗

Apoptosis in neonatal murine lung exposed to hyperoxia.

Exposure to high concentrations of oxygen in the neonatal period may impair lung growth and is a major contributing factor to the development of bronchopulmonary dysplasia. Cell death from hyperoxic injury may occur through either an apoptotic or nonapoptotic pathway, and we were interested in determining the type of cell death that occurs in the lung of neonatal mice exposed to hyperoxia. We found increased levels of Bax messenger RNA, a gene associated with apoptosis, in the lungs of neonatal mice born and raised in 92% hyperoxia. We next determined the extent of apoptosis taking place in the lungs of neonatal mice exposed to hyperoxia using terminal deoxyribonucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick-end labeling in 3.5-, 4.5-, and 5.5-d-old neonatal lung. The number of apoptotic cells in peripheral lung was significantly higher in the 3.5-, 4.5-, and 5.5-d-old mice treated with oxygen compared with that in the room-air control mice. Further, the number of apoptotic cells in the lung increased with longer exposure duration. In murine lung bronchus cells exposed to hyperoxia, growth arrest occurred after 48 h of oxygen exposure. Using annexin V binding, necrotic cell death was found to be the major form of cell death in these cells after 72 h of hyperoxic exposure. We conclude that 92% hyperoxia causes significant lung injury in neonatal mice exposed to hyperoxia, and that the number of apoptotic cells in the lung increases the longer the duration of exposure. The increase in apoptosis from hyperoxic exposure during a critical period of lung development may be an important factor in the impaired lung growth and remodeling that occur in animals exposed to high oxygen concentrations. Finally, it appears that hyperoxic injured cells in neonatal lung undergo both apoptotic and nonapoptotic cell death.

Animals↗

Increased level of HSP27 but not of HSP72 in human heart allografts in relation to acute rejection.

BACKGROUND: Increased expression of heat shock proteins (HSPs) was assumed during cardiac allograft rejection. To find evidence for this in man, we quantified HSP27 and HSP72 in cardiac allograft biopsies. METHODS: In parallel to histological assessment of rejection, HSP27 was quantified by Western blotting in a total of 43 biopsies sampled from 3 patients. HSP72 was analyzed in parallel in 30 of the 43 cases. For comparison, HSPs were analyzed in myocardium. RESULTS: HSP27 was significantly higher in rejecting cardiac allografts than in non-rejecting allografts and non-failing myocardium (1.52 +/- 0.25 vs. 0.83 +/- 0.11 vs. 0.50 +/- 0.05 microg/mg protein). Similarity for HSP72 (6.27 +/- 1.54 vs. 4.06 +/- 1.03 vs. 6.27 +/- 0.76 microg/mg protein) was not found. CONCLUSION: For the first time in humans with cardiac allograft rejection, increased expression of HSP27, which could be important for cardiac self-protection, was demonstrated. For the lack of increased HSP72 expression, the influence of the cyclosporine A treatment was discussed.

Acute Disease↗

Novel interaction between the transcription factor CHOP (GADD153) and the ribosomal protein FTE/S3a modulates erythropoiesis.

The transcription factor CHOP (GADD153) heterodimerizes with other C/EBP family members, especially C/EBPbeta, thus preventing their homodimerization and binding to DNA sequences specific for the homodimers. Some CHOP-C/EBP heterodimers apparently bind to alternative DNA sequence and thereby regulate the transcription of other genes. Recently, we demonstrated that CHOP is up-regulated during certain stages of erythroid differentiation and that ectopic overexpression of CHOP enhances this process (Coutts, M., Cui, K., Davis, K. L., Keutzer, J. C., and Sytkowski, A. J. (1999) Blood 93, 3369-3378). In the present study, we report that CHOP also interacts with another non-C/EBP protein designated v-fos transformation effector (FTE) (Kho, C. J., and Zarbl, H. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 2200-2204), which is identical to ribosomal protein S3a (Metspalu, A., Rebane, A., Hoth, S., Pooga, M., Stahl, J. , and Kruppa, J. (1992) Gene (Amst.) 119, 313-316). Bacterially expressed His-CHOP and in vitro translated (35)S-labeled FTE/S3a-Gal4 fusion protein co-immunoprecipitated using anti-CHOP antibodies, and both anti-CHOP and anti-FTE/S3a antibodies co-immunoprecipitated CHOP and FTE/S3a from lysates of Rauscher murine erythroleukemia cells overexpressing both proteins. The in vivo interaction of CHOP and FTE/S3a was also demonstrated in cells overexpressing FTE/S3a but with endogenous expression levels of CHOP. Western blot analysis demonstrated co-localization of CHOP and FTE/S3a in both the cytosol and the nuclei of non-transfected cells. Overexpression of FTE/S3a inhibited differentiation of Rauscher cells induced either by erythropoietin or by dimethyl sulfoxide. This inhibition was reversed partially by simultaneous overexpression of CHOP or of antisense fte/S3a. FTE/S3a appears to be a bifunctional ribosomal protein that regulates CHOP and, hence, C/EBP function during erythropoiesis.

Animals↗

Regulation of drug sensitivity by ribosomal protein S3a.

When bcl-2 is immunoprecipitated from (32)P-labeled cell extracts of all-trans retinoic acid (ATRA)-treated acute myeloblastic leukemia (AML) blasts, a phosphorylated protein of approximately 30 kd is coprecipitated. This protein has been identified as ribosomal protein S3a. The biologic effects of S3a include favoring apoptosis and enhancing the malignant phenotype. We sought to determine whether S3a, like bcl-2, influenced the response of cells to chemotherapeutic drugs and ATRA. Cell lines were studied in which S3a was genetically increased or disrupted; increased S3a was regularly associated with increased plating efficiency and increased sensitivity to either cytosine arabinoside (ara-C) or doxorubicin (DNR). S3a did not affect the sensitivity of cells to paclitaxel. Pulse exposures to either (3)HTdR or ara-C showed a greater percentage of clonogenic cells in the S phase of the cell cycle in cells with increased S3a than in controls. Cells with increased S3a responded to ATRA by increased ara-C or DNR sensitivity, whereas cells with reduced S3a protein were either protected by ATRA or not affected. We studied cryopreserved blast cells from patients with AML or chronic myelomonocytic leukemia (CMML). S3a protein levels were heterogeneous in these populations. In 32 cryopreserved blast populations, S3a levels were significantly correlated with both bcl-2 and with cell growth in culture. As in cell lines, high S3a in cryopreserved blasts was associated with ATRA-induced sensitization to ara-C. No significant association was seen between S3a levels and response to treatment.

Animals↗

Effect of corneal thickness on the accuracy of intraocular pressure measurement in rabbits after excimer laser photoablation.

PURPOSE: To determine whether intraocular pressure (IOP) measurements using pneumotonometry and Tono-Pen tonometry are accurate after excimer laser photoablation of the central cornea in rabbits. SETTING: Department of Ophthalmology, Scott & White Hospital, Texas A & M University College of Medicine, Temple, Texas, USA. METHODS: Ten rabbits had excimer laser phototherapeutic keratectomy (PTK) in the right eye to thin the central corneal thickness (CCT) by approximately 20%. The left eye served as the control. The diameter of each PTK was 6.0 mm. Four weeks later, the rabbits were examined under general anesthesia. The eyes were cannulated, and IOP was maintained at 10, 20, and 30 mm Hg and measured using an IOP transducer, Tono-Pen, and pneumotonometer at each pressure level. The readings were compared. RESULTS: One rabbit was excluded from the study because of recurrent erosions and corneal edema. There was no statistically significant difference in measurement error between the operated eye and control eye for the Tono-Pen (P = .12) or pneumotonometer (P = .35). There were significant differences in measurement error as pressure increased for the Tono-Pen (P < .01) and pneumotonometer (P < .01). The increase in measurement error with the increase in pressure was similar in the 2 eyes: Tono-Pen (P = .96) and pneumotonometer (P = .55). A paired t test showed the observed CCT was comparable (P = .79) in the pre-laser right eyes (mean 400.56 microm +/- 11.29 [SD]) and control eyes (mean 401.78 +/- 19.78 microm). CONCLUSION: Photoablation of approximately 20% of the CCT in rabbits by excimer laser PTK did not significantly alter the accuracy of IOP measurements by the Tono-Pen or pneumotonometer.

Animals↗

Corneal subepithelial nodular scarring treated with phototherapeutic keratectomy in a child with Rothmund-Thomson syndrome.

PURPOSE: To report the use of excimer laser phototherapeutic keratectomy (PTK) in the treatment of corneal subepithelial nodular scarring in a child with Rothmund-Thomson syndrome (RTS), a form of ectodermal dysplasia. METHODS: A case report and review of the literature. RESULTS: Excimer laser PTK successfully treated the recurrent anterior stromal fibrosis and irregular astigmatism, facilitating visual development. The use of excimer laser PTK prevented the development of dense amblyopia in this child. CONCLUSION: This case demonstrates an indication for excimer laser PTK in the pediatric population.

Child, Preschool↗

Mismatch repair proteins and microsatellites hit clinical practice.

Hereditary nonpolyposis colon cancer (HNPCC) is one of the most common familial cancers with characteristic molecular changes that are different from those found in familial adenomatous polyposis (FAP) coli. Genetic mutations in the germline and somatic cells lead to loss of expression of one of the two most commonly involved mismatch repair genes, hMSH2 or hMLH1, and consequently, to expansion of certain repetitive DNA sequences (microsatellite instability (MSI)). The paper describes a distinct subtype of "HNPCC-like" sporadic colonic carcinoma that can easily be identified by immunohistochemistry. Recognition of this subtype of colonic cancer is important because it occurs in the younger age group and is associated with better survival, but also a five-fold chance of developing a second colorectal carcinoma compared to "conventional" colorectal carcinomas.

Adaptor Proteins, Signal Transducing↗

Biopsy diagnosis of malignant versus benign liver "nodules": new helpful markers. An update.

Improved imaging techniques have led to an increased detection of suspicious "nodules" in the cirrhotic and noncirrhotic liver. Although the histologic diagnosis of clearly benign or clearly malignant lesions is usually straightforward, problems arise in the differential diagnosis of benign "nodules" and dysplastic lesions or well-differentiated hepatocellular carcinomas. This is especially so in limited diagnostic material, such as cytologic preparations from fine-needle aspirates and needle-core tissue biopsies. Recently, additional helpful markers have been described that may help to establish a conclusive diagnosis even on such material. Two of these, the reticulin stain and immunohistochemical staining for CD34, have been found to be useful also in cytologic cellblock material. Telomerase, a complex enzyme that is active in most malignant tumors, has also been found to show strong activity in most hepatocellular carcinomas. Mention is made of an immunohistochemical marker (MOC-31), reported to be useful in the differentiation between metastatic adenocarcinoma and hepatocellular carcinoma.

Adenocarcinoma↗

Preparation of response force and movement direction: onset effects on the lateralized readiness potential.

Two experiments assessed the preparatory effects of advance information about response force and movement direction on the lateralized readiness potential (LRP). In a choice reaction time (RT) task, an imperative stimulus required an isometric flexion or an extension of the left or right index finger. Prior information about response force or about movement direction reduced RT and shortened the interval from the onset of the imperative stimulus up to the onset of the LRP. Advance information, however, about direction but not about force decreased the interval from LRP onset to the onset of the overt response. The identical pattern of results was obtained in a second experiment, in which each participant performed both precue conditions. The findings of both experiments support the notion that response force is specified before movement direction. These results are consistent with the view accordingly different mechanisms are involved in the specification of muscle force and movement direction.

Adult↗