Primary squamous cell carcinoma of endometrium: a case report.

A case of primary squamous cell carcinoma of the endometrium (PSCCE) in a virgin treated with surgery--abdominal hysterectomy and bilateral salpingo-oophorectomy, and followed by whole pelvic radiation is presented. The tumor recurred 12 months later and the patient then underwent relaparotomy and excision of recurrent tumor of the pelvis; right hemicolectomy and rectosigmoidectomy were done due to recurrence and metastasis of the primary tumor. After re-operation, six courses of cisplatin and 5-fluorouracil were given intravenously for cytotoxic effects at 3-week intervals. The patient's disease progressed despite therapy, and she died three months after the last cycle or 21 months after the first diagnosis was made.

Adnexal masses in pregnancy: a review of eight cases undergoing surgical management.

OBJECTIVE: Our purpose was to determine maternal and foetal outcome in patients undergoing surgery for a pelvic mass in pregnancy. STUDY DESIGN: Maternal and foetal records (outcomes) of eight cases of adnexal masses associated with intrauterine pregnancy that required laparotomy or aspiration or that were diagnosed incidentally at the time of caesarean section were reviewed. The review was performed on patients who were seen with an adnexal mass in pregnancy from January 1994 to February 2001. We included patients with simple or complex masses > or = 6 cm that were persistent on ultrasonographic evaluation and patients with adnexal masses with complications (torsion, haemorrhage). We excluded cysts that spontaneously resolved by 16 weeks' gestation. RESULTS: Eight patients of 16,472 deliveries were identified with adnexal masses that satisfied the above criteria. Six patients underwent laparotomy in the first and/or the second trimester of pregnancy. In two of them emergency laparotomy were done due to torsion or haemorrhage as a complication of the adnexal masses. In all patients benign ovarian tumors were found. Two patients underwent transvaginal aspiration of simplex cysts due to subtorsion in the first trimester of gestation (negative results on cytological study). All of these eight patients had term deliveries. Two patients, due to obstetrical reasons, underwent caesarean section. CONCLUSION: The incidence of an adnexal mass during pregnancy in our population is consistent with what has been reported in the literature. We emphasize that transvaginal aspiration and drainage of symptomatic simplex cysts in the first trimester and percutaneous cysts in the second trimester can avert laparotomy. Our data support a randomised clinical study to determine optimal management of an adnexal mass in pregnancy.

Structure of human dipeptidyl peptidase I (cathepsin C): exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases.

Dipeptidyl peptidase I (DPPI) or cathepsin C is the physiological activator of groups of serine proteases from immune and inflammatory cells vital for defense of an organism. The structure presented shows how an additional domain transforms the framework of a papain-like endopeptidase into a robust oligomeric protease-processing enzyme. The tetrahedral arrangement of the active sites exposed to solvent allows approach of proteins in their native state; the massive body of the exclusion domain fastened within the tetrahedral framework excludes approach of a polypeptide chain apart from its termini; and the carboxylic group of Asp1 positions the N-terminal amino group of the substrate. Based on a structural comparison and interactions within the active site cleft, it is suggested that the exclusion domain originates from a metallo-protease inhibitor. The location of missense mutations, characterized in people suffering from Haim-Munk and Papillon-Lefevre syndromes, suggests how they disrupt the fold and function of the enzyme.

A TOPRIM domain in the crystal structure of the catalytic core of Escherichia coli primase confirms a structural link to DNA topoisomerases.

Primases synthesize short RNA strands on single-stranded DNA templates, thereby generating the hybrid duplexes required for the initiation of synthesis by DNA polymerases. We present the crystal structure of the catalytic unit of a primase enzyme, that of a approximately 320 residue fragment of Escherichia coli primase, determined at 2.9 A resolution. Central to the catalytic unit is a TOPRIM domain that is strikingly similar in its structure to that of corresponding domains in DNA topoisomerases, but is unrelated to the catalytic centers of other DNA or RNA polymerases. The catalytic domain of primase is crescent-shaped, and the concave face of the crescent is predicted to accommodate about 10 base-pairs of RNA-DNA duplex in a loose interaction, thereby limiting processivity.

Revised definition of substrate binding sites of papain-like cysteine proteases.

A review of kinetic and structural data has enabled us to reconsider the definition of substrate binding sites in papain-like cysteine proteases. Only three substrate binding sites, S2, S1 and S1', involve main as well as side chain contacts between substrate and enzyme residues. Interactions between the enzymes and the substrate P3 and P2' residues are based on side chains (an exception is cathepsin B which is a carboxydipeptidase), so their interaction surface spreads over a relatively wide area. The location and definition of substrate binding sites beyond S3 and S2' is even more questionable.

Crystal structure of porcine cathepsin H determined at 2.1 A resolution: location of the mini-chain C-terminal carboxyl group defines cathepsin H aminopeptidase function.

BACKGROUND: Cathepsin H is a lysosomal cysteine protease, involved in intracellular protein degradation. It is the only known mono-aminopeptidase in the papain-like family and is reported to be involved in tumor metastasis. The cathepsin H structure was determined in order to investigate the structural basis for its aminopeptidase activity and thus to provide the basis for structure-based design of synthetic inhibitors. RESULTS: The crystal structure of native porcine cathepsin H was determined at 2.1 A resolution. The structure has the typical papain-family fold. The so-called mini-chain, the octapeptide EPQNCSAT, is attached via a disulfide bond to the body of the enzyme and bound in a narrowed active-site cleft, in the substrate-binding direction. The mini-chain fills the region that in related enzymes comprises the non-primed substrate-binding sites from S2 backwards. CONCLUSIONS: The crystal structure of cathepsin H reveals that the mini-chain has a definitive role in substrate recognition and that carbohydrate residues attached to the body of the enzyme are involved in positioning the mini-chain in the active-site cleft. Modeling of a substrate into the active-site cleft suggests that the negatively charged carboxyl group of the C terminus of the mini-chain acts as an anchor for the positively charged N-terminal amino group of a substrate. The observed displacements of the residues within the active-site cleft from their equivalent positions in the papain-like endopeptidases suggest that they form the structural basis for the positioning of both the mini-chain and the substrate, resulting in exopeptidase activity.

Crystal structure of the wild-type human procathepsin B at 2.5 A resolution reveals the native active site of a papain-like cysteine protease zymogen.

The structure of the wild-type human procathepsin B has been refined to a crystallographic R-value of 0.18 and R-free of 0.23 exploiting the data obtained from new crystals that diffract beyond 2.5 A resolution. The structure confirms two previously presented, lower-resolution structures. The structure of the propeptide chain folds on the surface of the enzyme domains and blocks access of substrate to the already formed active site. Abundant solvent molecules fill the cavities between the propeptide and the enzyme part of the molecule. The propeptide structure is compared with a substrate model in the S2, S1, S1' and S2' binding sites. In this crystal form the cathepsin B occluding loop residues adopt yet another conformation. The structures show that the occluding loop region between the residues Cys108 and Cys119 behaves quite independently from the rest of the structure and easily adapts to changes in environment. The variety of the observed conformations of the occluding loop is in agreement with other data showing that the loop is responsible for limiting cathepsin B activity to that of a carboxydipeptidase. The region before Cys108 is essentially the same as in the mature structure, whereas the region from Cys119 to Thr125 is raised compared to the mature form by the propeptide squeezed between it and the enzyme domains, surface. The structure strongly suggests that processing of procathepsin B during its autoactivation is not unimolecular.

Transabdominal chorionic villus sampling in the second and third trimesters of high-risk pregnancies.

Late chorionic villus sampling (placental biopsy) under ultrasound guidance was carried out in 800 (80 per cent) cases in the second trimester and 200 (20 per cent) cases in the third trimester of pregnancy. Out of 1000 placental biopsies, 250 (25 per cent) were performed because of suspicious ultrasonographic findings. Colour Doppler was used to investigate the uteroplacental and fetal vessels in 300 (30 per cent) pregnancies before and after late chorionic villus sampling (CVS). In the same group, mean serum alpha-fetoprotein (AFP) levels increased after sampling in 20 (6.7 per cent) patients. In 20 patients (2 per cent), complications between sampling and delivery were found. A placental haematoma measuring 0.5-1 ml was seen at the sampling site in 4 (0.4 per cent) patients in the second trimester of pregnancy and in 3 (0.3 per cent) in the third trimester. Two (0.2 per cent) demonstrated fever, but there were no instances of chorioamnionitis. There were only three (0.3 per cent) spontaneous abortions 4-6 weeks after late CVS. However, there was no correlation between AFP elevation, placental haematoma, Doppler measurements, and spontaneous abortion. Cytogenetic findings were obtained in 990 (99 per cent) of 1000 placental samplings. We found 60 (6.0 per cent) chromosomal abnormalities. In the group with suspicious ultrasonic findings (250 cases), we found significant oligohydramnios in 125 (50 per cent) and significant polyhydramnios in 60 (24 per cent), and 45 (18 per cent) had chromosomal abnormalities. Among the 60 patients with chromosomal abnormalities, ultrasonographic findings in 10 (16.7 per cent) were detected after the 20th week of pregnancy. There were no significant differences in mean pulsatility index (PI) in the uteroplacental and fetal vessels before and after late CVS. Preliminary data from five trisomic fetuses (three trisomy 21 and two trisomy 18) showed abnormally increased umbilical PI and abnormally decreased middle cerebral artery PI.

Ultrasonic fetal and placental tissue characterization and lung maturity.

OBJECTIVE: To determine a relationship between gestational age and the quantitative assessment of ultrasonic signs of placental tissue, fetal lung and liver tissue for determining fetal lung maturity in normal pregnancies and pregnancies with preeclampsia. METHODS: Placental, fetal lung and fetal liver tissue was examined by ultrasound in 240 normal and 60 preeclamptic pregnancies at 30-41 weeks' gestation. All patients underwent ultrasonically guided amniocentesis to obtain the lecithin-sphingomyelin ratio. The placentas of 160 patients after delivery were placed in water at body temperature for ultrasonic echo amplitude analysis. The coefficients of variation (the standard deviation divided by the mean value) of gray levels of the pixels in the region of interest obtained from images of the placenta, fetal liver and lung, were used to characterize the tissue in different groups during pregnancy. RESULTS: The coefficients of variation in mature fetuses were > 29% for placentas in vivo, > 34% for placentas in vitro, > 28% for liver tissue and > 30% for lung tissue. In mature fetuses the ratio of coefficients of variation of placental tissue in vivo against placental tissue in vitro was > 0.80, placental tissue in vivo against lung tissue > 0.90, lung tissue against liver tissue > 1.10 and placental tissue in vivo against liver tissue > 1.00. CONCLUSION: The placental and fetal lung tissue of preeclamptic patients tended to have higher coefficients of variation throughout pregnancy. These results were significantly higher when associated with low-birth-weight babies. There were no significant differences in fetal liver tissue between normotensive and preeclamptic groups.

Crystal structures of human procathepsin B at 3.2 and 3.3 Angstroms resolution reveal an interaction motif between a papain-like cysteine protease and its propeptide.

A wild-type human procathepsin B was expressed, crystallized in two crystal forms and its crystal structure determined at 3.2 and 3.3 Angstroms resolution. The structure reveals that the propeptide folds on the cathepsin B surface, shielding the enzyme active site from exposure to solvent. The structure of the enzymatically active domains is virtually identical to that of the native enzyme [Musil et al. (1991) EMBO J. 10, 2321-2330]: the main difference is that the occluding loop residues are lifted above the body of the mature enzyme, supporting the propeptide structure.

Crystal structure of cathepsin B inhibited with CA030 at 2.0-A resolution: A basis for the design of specific epoxysuccinyl inhibitors.

Crystals of cysteine protease human cathepsin B inhibited with CA030 (ethyl ester of epoxysuccinyl-Ile-Pro-OH) [Murata, M., et al. (1991) FEBS Lett. 280, 307-310; Towatari, T., et al. (1991) FEBS Lett. 280, 311-315] were isomorphous to a previous published structure of cathepsin B [Musil, D., et al. (1991) EMBO J. 10, 2321-2330]. The crystal structure of the complex was refined at 2.0-A resolution to an R-value of 0.194. CA030 is well-defined in the electron density. The Ile-Pro-OH part of CA030 mimics a substrate P1' and P2' residues. The structure thus reveals for the first time a substratelike interaction in the S1' and S2' sites of a papain-like cysteine protease. The CA030 ethyl ester group occupies the S2 site. The structure confirms the role of residues His 110 and His 111 as the receptors of a peptidic substrate C-terminal carboxylic group. The structure suggests that an epoxysuccinyl fragment can be used to extend binding into primed and nonprimed substrate binding sites of a papain-like cysteine protease.

First trimester diagnosis of cystic hygromata using transvaginal ultrasound and cytogenetic evaluation.

We studied the outcome of fetuses in whom cystic hygroma was diagnosed in the first and early second-trimester of pregnancy using transvaginal ultrasonography. The purpose of this study was to evaluate the association between fetal cystic hygroma and fetal cytogenetic abnormalities, and the long-term prognosis. Thirty-five consecutive fetuses between 9.1 and 13.4 weeks of gestation diagnosed as having a nuchal hygroma were evaluated ultrasonographically and karyotyped. Those with a normal chromosome complement were ultrasonographically monitored throughout the remainder of the pregnancy to document the resolution of the hygroma. Eighteen of thirty-five fetuses were found to have a normal karyotype and five of these were aborted electively. The hygromas resolved in ten of these karyotypically normal fetuses within four weeks of initial diagnosis and they were phenotypically normal at birth. Seventeen fetuses were karyotypically abnormal with trisomy twenty-one being the most common abnormality. Prenatal cytogenetic analysis should be offered to women with fetal cystic hygroma diagnosed in the first trimester. A normal outcome is likely in those without chromosome abnormalities.

Preoperative diagnosis of primary fallopian tube carcinoma by transvaginal ultrasound, cytological finding and CA-125.

Primary Fallopian tube carcinoma is rarely diagnosed preoperatively. We present the case of a 69-year-old woman with primary tubal carcinoma, which was diagnosed preoperatively on the basis of the cytological finding, characteristic features on transvaginal sonography, transvaginal color flow imaging and elevated CA-125. Transvaginal color Doppler imaging demonstrated the tumor revealed areas of neovascularization with characteristic low impedance (resistance index, 0.34 and pulsatility index, 0.62). Pathohistologic confirmation of the clearcell carcinoma has been done.

Bovine cathepsins S and L: isolation and amino acid sequences.

The purification procedure of cathepsin S includes acid activation of spleen homogenate, incubation at 37 degrees C, precipitation with (NH4)2SO4 in H2O/tert-butanol medium, gel chromatography, chromatofocusing, covalent chromatography and cation chromatography of FPLC system. Cathepsin S has a M(r) of about 24,000 Da with pI of 6.5 and 6.8. The mixture of both forms gave a single sequence. Cathepsin L was purified from bovine kidney by acid treatment and incubation of 37 degrees C, precipitation by (NH4)2SO4, two ion exchange chromatographies on CM-Sephadex, gel chromatography and ion exchange chromatography on FPLC system. Cathepsin L exists in multiple forms with pI 5.3-5.7 and M(r) of about 29,000 Da. N-terminal amino acid sequence confirms that cathepsin L and cathepsin S are different enzymes.

First-trimester diagnosis of cystic hygromata by transvaginal sonography.

This paper reports two cases of fetal nuchal cystic hygromata diagnosed in the first trimester of pregnancy by transvaginal sonography. In the first case, at 10 weeks' gestation, in addition to the hygromata, the fetus had an exomphalos. In the second case, at 13 weeks' gestation, the fetus had hydrocephalus and pleural effusion. Transabdominal chorion villus sampling was carried out and the fetal karyotypes were trisomy 18 and trisomy 21, respectively.

The complete amino acid sequence of bovine cathepsin S and a partial sequence of bovine cathepsin L.

The complete amino acid sequence of bovine spleen cathepsin S has been determined. The single-chain protein contains 217 residues and has a Mr of 23,682. The primary structure was determined by sequencing of native protein and the peptides obtained by proteolytic cleavage with beta-trypsin, papaya proteinase IV and by chemical cleavage with cyanogen bromide. Comparison of the amino terminal sequences of the heavy and the light chain of bovine cathepsin L with that of bovine cathepsin S clearly indicates that the enzymes are structurally different.

Transabdominal placental biopsy in the second and third trimester of pregnancy.

Transabdominal placental biopsy under ultrasound guidance was carried out in 260 cases in the second trimester and 50 cases in the third trimester of pregnancy. Placental tissue was aspirated using an 18 or 20 gauge needle. In a total of 310 placental biopsies in the second and third trimester, 100 were performed because of suspicious ultrasonographic findings. Placental biopsy is simple in the presence of severe oligohydramnios where fetal blood sampling is usually more difficult. Oligohydramnios and polyhydramnios were the ultrasonographic findings in 50% of cases and were found to be associated with 30% of abnormal chromosomal findings. There was one (0.3%) abortion within two weeks following placental biopsy. Placental biopsy did not affect the outcome of the pregnancy.

Ultrasonography in the detection of cervical incompetency.

In 80 pregnancies with clinical and ultrasonic signs of cervical incompetency, the length of the cervix and the thickness of the anterior wall of a lower uterine segment have been evaluated ultrasonically. We have also measured the width of the endocervical canal and studied the prolapse of fetal membranes (with fetal parts) into the endocervical canal. We evaluated these same parameters in 80 healthy pregnancies. The length of the cervix, the thickness of the anterior wall of a lower uterine segment, and the width of the endocervical canal were followed longitudinally in the patients from the 10th to the 36th gestation week. No statistically significant differences between age groups were found. In four age groups at risk for cervical incompetency, cervical lengths and wall thickness were significantly different (p less than 0.001) from those in comparable controls. Forty-five percent of the patients in the at-risk group, with cervical cerclage, delivered at 37.3 (range: 32 to 41) weeks and 6.25% of pregnancies ended in abortion when the amniotic membrane herniated into the cervical canal, with or without some part of the fetus.