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Biomedical subjects

C Stephan

Publications and source records attributed to C Stephan.

At least 55 records · Page 3Linked to original sources
PubMed2000

Molecular forms of prostate-specific antigen and human kallikrein 2 as promising tools for early diagnosis of prostate cancer.

Prostate-specific antigen (PSA) is the most useful marker in the early detection of prostate cancer and for the monitoring of patients with this diagnosis. Molecular forms of PSA and also human kallikrein 2 have been used to discriminate between benign prostatic hyperplasia and prostate cancer as well as for the detection of prostate cancer within the gray zone of PSA. In this respect, a literature survey on the diagnostic validity of free PSA (fPSA) related to total PSA (tPSA), PSA bound to alpha1-antichymotrypsin (ACT-PSA), and complexed PSA is given together with our results. The ratio of fPSA:tPSA has been shown to improve the specificity of prostate cancer diagnosis on the basis of tPSA measurements. Unnecessary biopsies can be reduced by about 19-64% in the total PSA range of 4-10 microg/liter while only missing 5-10% of cancers. Furthermore, carcinomas in patients with PSA values <4 microg/liter can be detected, indicating an improved sensitivity because of the percent fPSA at low PSA values. ACT-PSA or complexed PSA alone and the calculated derivatives are not superior in their discriminatory power compared with the percent fPSA. The diagnostic significance of the other molecular PSA forms and human kallikrein 2 needs to be evaluated in more extensive clinical trials.

Biomarkers, Tumor↗
PubMed1999

The mechanical effect of blocking screws ("Poller screws") in stabilizing tibia fractures with short proximal or distal fragments after insertion of small-diameter intramedullary nails.

OBJECTIVES/HYPOTHESIS: To evaluate the mechanical effects of medial and lateral blocking screws in supplementing intramedullary nail fixation of high proximal and low distal tibial fractures treated with small-diameter intramedullary nails. STUDY DESIGN: Intact fresh human cadaveric tibiae were sectioned to provide ten distal segments measuring seventy millimeters and ten proximal segments measuring ninety millimeters. In the distal segments, stainless steel solid eight-millimeter tibial nails were advanced to eight millimeters from the ankle joint. Two transverse and one anterior-posterior (AP) locking screw were inserted using a custom-made jig. The same jig was used for the placement of a medial and a lateral blocking screw (BS) in the AP direction, nine millimeters above the superior most interlocking screw and eight millimeters distal to the lower end of the segment. In the proximal segments, two interlocking screws (both static and dynamic screws) were placed in a medial-lateral direction with the use of the insertion handle. A jig was used for placement of a medial and a lateral BS in the AP direction, nine millimeters below the lower transverse interlocking screw and sixteen millimeters proximal to the lower end of the segment. The bone-implant construct (BIC) was embedded and fixed in a materials testing machine. The BICs were loaded in the medial-lateral direction at a distance of 185 millimeters from the nail ends with loads from -150 newtons to + 150 newtons. Force-displacement curves were recorded before and after insertion of the BSs. RESULTS: In proximal BICs, the addition of BSs decreased the deformation of the BICs 25 percent, from 8.9 +/- 1.9 degrees [mean +/- standard deviation (SD)] in the control group to 6.8 +/-1.1 degrees in the BS group (mean +/- SD) (p < 0.0001). In distal BICs, the addition of BSs decreased the deformation of the BICs 57 percent, from 9.5 +/- 1.4 degrees (mean +/- SD) in the control group to 4.0 +/- 1.0 degrees in the BS group (mean +/- SD) (p < 0.0001). CONCLUSIONS: The study suggests that medial and lateral blocking screws can increase the primary stability of distal and proximal metaphyseal fractures after nailing and can be an effective tool for selected cases that exhibit malalignment and/or instability.

Aged↗
PubMed1999

The use of Poller screws as blocking screws in stabilising tibial fractures treated with small diameter intramedullary nails.

Intramedullary nailing of metaphyseal fractures may be associated with deformity as a result of instability after fixation. Our aim was to evaluate the clinical use of Poller screws (blocking screws) as a supplement to stability after fixation with statically locked intramedullary nails of small diameter. We studied, prospectively, 21 tibial fractures, 10 in the proximal third and 11 in the distal third in 20 patients after the insertion of Poller screws over a mean period of 18.5 months (12 to 29). All fractures had united. Healing was evident radiologically at a mean of 5.4+/-2.1 months (3 to 12) with a mean varus-valgus alignment of -1.0 degree (-5 to 3) and mean antecurvatum-recurvatum alignment of 1.6 degrees (-6 to 11). The mean loss of reduction between placement of the initial Poller screw and follow-up was 0.5 degrees in the frontal plane and 0.4 degrees in the sagittal plane. There were no complications related to the Poller screw. The clinical outcome, according to the Karström-Olerud score, was not influenced by previous or concomitant injuries in 18 patients and was judged as excellent in three (17%), good in seven (39%), satisfactory in six (33%), fair in one (6%), and poor in one (6%).

Adolescent↗
PubMed1998

Identification of a human cDNA encoding a kinase-defective cdk5 isoform.

The cyclin-dependent kinase 5 (Cdk5) catalytic subunit is expressed in both cycling and noncycling cells and is present in many tissues. Neuronal and muscle cells contain the highest amount of this protein. The p35 protein, which is expressed solely in the brain, activates Cdk5. Cdk5 activity is involved in terminal differentiation of neurons and muscle cells. We attempted to clone cdk5 by PCR from a human fetal brain cDNA library. Surprisingly, we amplified two forms of the cdk5 gene, the wild type and a cdk5 variant that lacks the complete kinase domain VI. The variant is also found in SH-SY-5Y neuroblastoma cells but not in T-cells, HeLa cells, the thymus, and placental tissue. The protein encoded by the cdk5 variant, the Cdk5 isoform (Cdk5i), purifies with p35 when coexpressed in insect cells. The activity associated with the heterodimer Cdk5i/p35 is found to be appreciably weaker than the wild-type Cdk5/p35 kinase. Moreover, Cdk5i/p35 cannot autophosphorylate its two subunits as with Cdk5/p35. Interestingly, kinase-defective Cdk5i can abolish the activity of wild-type Cdk5 when both are coexpressed with p35 in insect cells, suggesting that Cdk5i may have a function in regulating Cdk5 activity in human cells too.

Amino Acid Sequence↗
PubMed1998

Relation of free PSA/total PSA in serum for differentiating between patients with prostatic cancer and benign hyperplasia of the prostate: which cutoff should be used?

A review on literature data is given concerning free prostate-specific antigen (f-PSA) and the corresponding cutoffs of f-PSA/t-PSA for differentiating patients with cancer of the prostate from those with benign prostatic hyperplasia. The special importance of the diagnostic criterion (sensitivity, specificity, efficiency) for establishing the cutoff is demonstrated. On the basis of our own data, the application of the f-PSA% is recommended as an additional decision criterion for biopsy.

Biopsy↗
PubMed1997

The influence of prostate volume on the ratio of free to total prostate specific antigen in serum of patients with prostate carcinoma and benign prostate hyperplasia.

BACKGROUND: Determining the ratio of free to total prostate specific antigen (f-PSA to t-PSA, calculated as the percentage of f-PSA [f-PSA%]) in serum allow for a clearer distinction between patients with prostate carcinoma (PCa) and patients with benign prostate hyperplasia (BPH) than determining the level of t-PSA alone. To find influencing factors on f-PSA%, the authors investigated prostate volume, TNM classification, and tumor stage. METHODS: The authors measured f-PSA and t-PSA in 36 men with untreated PCa (tumor classification: T1, 2, 3pNO, MO), 44 patients with BPH, and 54 healthy controls. Prostate volume was determined by transrectal ultrasound. RESULTS: The median values of t-PSA and f-PSA% were 7.8 micrograms/L and 10.5% in PCa patients, 4.3 micrograms/L and 20.8% in patients with BPH, and 1.4 micrograms/L and 23.6% in the control group. Patients with PCa had a significantly lower proportion of f-PSA than BPH patients and healthy men. There was no correlation of f-PSA% to TNM stage or tumor grade. In PCa patients a significant positive correlation (correlation coefficient [r] = 0.51, P < 0.001) was found between f-PSA% and prostate volume, whereas there was no significant correlation in BPH patients (r = -0.27, P > 0.05). There was a significant difference in f-PSA% between PCa and BPH patients with prostate volumes smaller than 40 cm3 (9.0% vs. 21.6%, P < 0.01) but not between patients in these 2 groups with prostate volumes exceeding 40 cm3 (15.1% vs. 18.2%, P = 0.11). CONCLUSIONS: Determining the ratio of f-PSA to t-PSA to discriminate between PCa and BPH patients yields significant results only in men with a prostate volume of less than 40 cm3.

Aged↗
PubMed1997

Elimination of serum free and total prostate-specific antigen after radical retropubic prostatectomy.

Elimination kinetics of serum total and free prostate-specific antigen were studied for a ten days course after radical retropubic prostatectomy on 11 patients suffering from organ confined prostate cancer. Samples were taken before operation, immediately after finishing the operation and 1, 2, 3, 4, 5, 6 h after prostatectomy and then once a day for the following ten days. The measurements were performed with AxSym assays from Abbott Laboratories. The elimination of both total and free prostate-specific antigen followed a biphasic kinetics. In the fast phase, the average of the individual elimination half-lives of total and free prostate-specific antigen amounted to 6.3 h (SD = 6.1 h; range: 0.55 to 37.1 h) and 0.57 h (SD = 0.18 h; range: 0.22 to 0.89 h), respectively. In the slow phase, total prostate-specific antigen disappeared with an average half-life of 85.6 h (SD = 11 h; range: 47.2 to 261.7 h) and free prostate-specific antigen with an average half-life of 14.4 h (SD = 10.4 h; range: 2.4 to 30.3 h). These results might be significant for the use of free and total prostate-specific antigen and its ratio as a diagnostic and prognostic tool.

Aged↗
PubMed1996

Role of mitochondria and C-terminal membrane anchor of Bcl-2 in Bax induced growth arrest and mortality in Saccharomyces cerevisiae.

In mammalian cells, the Bcl-2 and Bcl-x(L) proteins suppress programmed cell death whereas the topographically similar Bax protein accelerates the apoptotic process. Recently published data suggest that expression of the human Bax-alpha gene is lethal for the yeast Saccharomyces cerevisiae and that this toxicity can be overcome by co-expressing Bcl-2 or Bcl-x(L). Our findings corroborate these results. However, we find that although Bax induction invariably stops cell growth under all circumstances, it does not lead to death in 'petite' cells. Petites cannot respire because they lack functional mitochondria. It seems that in 'grande' cells, which do possess normal mitochondrial DNA, nutritional limitation is critical for increased mortality. Surprisingly, murine Bcl-2 lacking the membrane anchor of human Bcl-2 has no effect on grande cells, but can efficiently rescue petites in rich medium. It has been suggested that the C-terminal membrane anchor of human Bcl-2 may have a crucial role in rescuing apoptosis in mammalian cells. When murine Bcl-2 is fused to the membrane anchor of yeast mitochondrial Mas70 protein, the Bcl-2 variant mBcl-2-mma rescues not only petites but also grandes, just like human Bcl-x(L). The rescuing ability of Bcl-x(L), which contains its own membrane anchor, surpasses that of mBcl-2-mma. Our results indicate that the process involving Bax-induced growth inhibition followed by possible lethality, and the rescuing effect of Bcl-2 and Bcl-x(L) is linked to yeast mitochondrial function. We propose a model which is consistent with these observations.

Amino Acid Sequence↗
PubMed1996

Analytical performance and clinical validity of two free prostate-specific antigen assays compared.

We compared two recently introduced commercial assays (CanAg and Immulite) for measuring free prostate-specific antigen (f-PSA), total PSA (t-PSA), and the ratio of t-PSA/f-PSA (f-PSA%) in control materials and sera of 54 healthy men, 50 patients with benign prostatic hyperplasia (BPH), and 45 patients with prostate cancer (PCa). The lower detection limits for f-PSA were 0.038 microgram/L and 0.004 microgram/L for the CanAg and Immulite assays, respectively. The within-run and between-day precisions of the Immulite assay were < 5%; the CanAg assay showed a poorer precision. Whereas f-PSA values differed between controls and patients but not between BPH and PCa patients, the f-PSA% values were lower in PCa patients than in BPH patients and controls. The receiver-operating characteristic (ROC) curve showed an improved diagnostic power of f-PSA% compared with t-PSA to discriminate between BPH and PCa. Discrimination limits of 16% (CanAg assay), and 15% (Immulite assay) are recommended for f-PSA%.

Autoanalysis↗
PubMed1995

Only in the presence of immunophilins can cyclosporin and FK506 disrupt in vivo binding of calcineurin A to its autoinhibitory domain yet strengthen interaction between calcineurin A and B subunits.

The two immunosuppressants cyclosporin A (CsA) and FK506 exert their major therapeutic effect by inhibiting T-cell activation. It is believed that the drugs first bind to their cellular receptors, known as immunophilins, and then target the protein phosphatase calcineurin for inhibition. The catalytic activity of calcineurin is regulated by its autoinhibitory domain (AID) and by the calcium-binding proteins calcineurin B (CnB) and calmodulin. We have used the yeast two-hybrid system to show that AID, CnB and calmodulin can only bind to a truncated catalytic subunit of yeast calcineurin (i.e.,CnA1 delta), devoid of AID, but not to full-length CnA1. Both CsA and FK506 cause disruption of the CnA1 delta-AID interaction, whereas their presence permits CnA1 delta to bind more strongly to CnB. In contrast, the binding of CnA1 delta to calmodulin is unaffected by the immunosuppressants. Significantly, in the absence of its cognate cytosolic receptor, neither CsA nor FK506 inhibits or stimulates the CnA1 delta-AID, CnA1 delta-CnB interactions. These in vivo observations not only provide supportive evidence for the mechanism by which drug-receptor complexes could modulate calcineurin activity but also unveil the possibility of identifying novel immunophilin-independent calcineurin inhibitors which may disturb the association of CnA1 delta to AID.

Base Sequence↗
PubMed1995

The neuroendocrine protein 7B2 acts as a molecular chaperone in the in vitro folding of human insulin-like growth factor-1 secreted from yeast.

The neuroendocrine protein 7B2 prevents premature activation of PC2, an enzyme involved in the processing of prohormones in the secretory pathway. We inquired if this chaperone-like function encompasses a broader role for 7B2 in the folding of hormone-like proteins. As a test, the fate of misfolded human insulin-like growth factor-1 (IGF1) was assessed, in the presence and absence of 7B2. Most of the recombinant IGF1 molecules, secreted from yeast, are a conglomeration of inactive multimers which are either disulfide-linked or mere physical aggregates. We find that yeast-produced 7B2 influences the in vitro conversion of inactive molecules into active monomers. However, the amounts of disulfide-linked dimers remain unaffected during this conversion. Interestingly, both 7B2 and the molecular chaperone DnaK interact with IGF1 in the yeast two-hybrid system. Like DnaK, 7B2 also binds the tumor suppressor protein p53. Binding of DnaK to exposed epitopes of aggregated proteins is known to be a prerequisite for deaggregation. It is conceivable that 7B2 participates in an analogous manner in the dissociation of non-covalently linked multimers of IGF1. Our results indicate that 7B2 might find an application in the deaggregation of potentially useful therapeutic proteins.

Base Sequence↗
PubMed1995

A C-terminal domain, which prevents secretion of the neuroendocrine protein 7B2 in Saccharomyces cerevisiae, inhibits Kex2 yet is processed by the Yap3 protease.

Recent reports reveal that the C-terminal half of the neuroendocrine polypeptide 7B2 selectively inhibits and binds PC2, a mammalian prohormone converting enzyme that is homologous to the yeast pro-alpha-factor processing protease Kex2. During attempted secretion of the 185 amino-acid human 7B2 in Saccharomyces cerevisiae, we observe that the protein is mostly retained inside the cell. However a mutant polypeptide (7B2 delta 1), where the C-terminal 48 amino acids of 7B2 are deleted, is efficiently secreted. Two shorter C-terminal truncations either permit poor secretion or no secretion at all. Surprisingly, full-length 7B2 but not 7B2 delta 1 abolishes the catalytic activity of Kex2, indicating that C-terminal residues of 7B2 might also be important for inhibition of the yeast protease. When the KEX2 gene is disrupted, yeast cells unexpectedly secrete a 7B2 variant similar in size to 7B2 delta 1, suggesting involvement of the alternate yeast prohormone convertase Yap3 in processing. Secretion is enhanced by overexpression of Yap3 and by the presence of a Lys-Arg residue at the processing site of precursor 7B2. These results purport that, in neuroendocrine cells too, secretion of 7B2 could be mediated by a homologue of Yap3.

Aspartic Acid Endopeptidases↗
PubMed1995

The unfolded-protein-response element discriminates misfolding induced by different mutant pro-sequences of yeast carboxypeptidase Y.

The C-terminal region of the chaperone-like pro-sequence (py) of yeast carboxypeptidase Y (CPY) is suggested to be crucial for the folding of mature CPY. In order to study the influence of hydrophobic residues in this domain, a set of mutations have been introduced in py. Unexpectedly, only small amounts of CPY precursors are expressed when Leu108, at the C-terminal end of py, is substituted for polar residues Lys, Arg or Asp. In contrast, substitution with hydrophobic residues Val, Ile or Ala permit normal expression. Interestingly, the poorly expressed molecules are coreglycosylated, implying that they have failed to leave the endoplasmic reticulum (ER). The ER-retained molecules caused an induction in the levels of BiP, signifying that polar substitutions at position 108 of pre-py-CPY induce misfolding. Quite surprisingly, a reporter gene, linked to concatamerized unfolded-protein-response elements, reveals that py-mediated misfolding of CPY is not really identical in all mutants. This shows that a simple transcriptional assay can assess the subtleties of pro-sequence mediated protein folding in an eukaryotic cell.

Base Sequence↗
PubMed1992

A mutant Kex2 enzyme with a C-terminal HDEL sequence releases correctly folded human insulin-like growth factor-1 from a precursor accumulated in the yeast endoplasmic reticulum.

Mutations in the pro region of the yeast DNA hybrid of prepro-alpha-factor and human insulin-like growth factor-1 (IGF-1) cause the accumulation, in the yeast Saccharomyces cerevisiae, of an unglycosylated precursor protein where the pre sequence is missing. The prepro sequence of the prepro-alpha-factor consists of a pre or signal sequence and a proregion which possesses three sites for N-glycosylation. Isolation of a precursor, where the pro region is still linked to IGF-1 through a pair of dibasic amino acid residues, implies that the polypeptide may have translocated into the endoplasmic reticulum (ER) but has not been processed by the Golgi membrane-bound Kex2 endoprotease. However, the lack of any N-glycosylation in the translocated polypeptide is surprising. The mutated pro region, can be processed, in vitro, by treatment with a soluble form of the Kex2 enzyme. It is also possible to release the pro region, in vivo, by coexpressing a mutant Kex2 protease which is partially retained in the ER with the help of the C-terminal tetrapeptide sequence, HDEL. The mature IGF-1, which is secreted from the intracellular pool of precursor proteins, is predominantly an active, monomeric molecule, corroborating observations that early removal of the pro region before folding in the ER helps to prevent aberrant intermolecular disulfide-bond formation in IGF-1. These results have revealed the utility of the ER-retained Kex2 enzyme as a novel in vivo biochemical tool.

Amino Acid Sequence↗