Incontinence,bladder neck mobility,and sphincter ruptures in primiparous women

Objective

To compare the function of the pelvic floor in primiparae before and during pregnancy with the status post partum concerning symptoms of incontinence, sphincter ruptures, bladder-neck mobility and the influence of the different modes of deliveries.

Methods

Questionnaire evaluating symptoms of urinary and anal incontinence in nulliparous women before and after delivery and correlating these symptoms with functional changes of the pelvic floor based on a careful gynaecologic examination as well as perineal and endoanal ultrasound.

Results

112 women were included in our study and came for the first visit, 99 women returned for follow-up 6 months after childbirth. Stress and flatus incontinence significantly increased from before pregnancy (3 and 12%) to after childbirth (21 and 28%) in women with spontaneous delivery or vacuum extraction. No new symptoms occurred after c-section. There was no significant difference between the bladder neck position before and after delivery. The mobility of the bladder neck was significantly higher after vaginal delivery using a vacuum extraction compared to spontaneous delivery or c-section.The bladder neck in women with post partum urinary stress incontinence was significantly more mobile than in continent controls. The endoanal ultrasound detected seven occult sphincter defects without any correlation to symptoms of anal incontinence.

Conclusion

Several statistically significant changes of the pelvic floor after delivery were demonstrated. Spontaneous vaginal delivery or vacuum extraction increases the risk for stress or anal incontinence, delivery with vacuum extraction leads to higher bladder neck mobility and stress incontinent women have more mobile bladder necks than continent women.     

胸腔镜治疗贲门失弛症

胸腔镜治疗贲门失弛症何启才ＢｒｙｎｅｒＵＭ王永清郦志军章文晓何正富作者单位：３１００１６浙江医科大学附属邵逸夫医院心胸外科（何启才、王永清、郦志军、章文晓、何正富）；美国罗马琳达大学医学中心外科（ＢｒｙｎｅｒＵＭ）１９９６年２月至４月我们应用胸腔镜行...     

Inflammatorischer, myofibroblastischer Tumor des Pankreas mit Beteiligung region?rer Lymphknoten

Inflammatorische myofibroblastische Tumoren (IMT) kommen überall im menschlichen K?rper vor. Aufgrund ihrer heteromorphen, mesenchymalen Morphologie ist die Abgrenzung dieser meist gutartigen Tumoren gegenüber Sarkomen problematisch, zumal sie auch infiltrativ destruierend wachsen und gelegentlich rezidivieren k?nnen. Die Manifestation im Pankreas ist au?erordentlich selten. Klinik und bildgebende Diagnostik lassen in der Regel an ein Malignom denken. Erst anhand der Histomorphologie kann die definitive Diagnose gestellt werden. Es wird der Fall einer 62-j?hrigen Patientin mit einem IMT des Pankreas beschrieben. über die Einbeziehung region?rer Lymphknoten in den inflammatorischen Prozess ist bei einem IMT des Pankreas bisher noch nicht berichtet worden.     

A novel structure involved in the formation of liver endothelial cell fenestrae revealed by using the actin inhibitor misakinolide

Hepatic endothelial fenestrae are dynamic structures that act as a sieving barrier to control the extensive exchange of material between the blood and the liver parenchyma. Alterations in the number or diameter of fenestrae by drugs, hormones, toxins, and diseases can produce serious perturbations in liver function. Previous studies have shown that disassembly of actin by cytochalasin B or latrunculin A caused a remarkable increase in the number of fenestrae and established the importance of the actin cytoskeleton in the numerical dynamics of fenestrae. So far, however, no mechanism or structure has been described to explain the increase in the number of fenestrae. Using the new actin inhibitor misakinolide, we observed a new structure that appears to serve as a fenestrae-forming center in hepatic endothelial cells.     

The involvement of CYP1A2 and CYP3A4 in the metabolism of clozapine

Aims Clozapine (CLZ), an atypical neuroleptic with a high risk of causing agranulocytosis, is metabolized in the liver to desmethylclozapine (DCLZ) and clozapine N-oxide (CLZ-NO). This study investigated the involvement of different CYP isoforms in the formation of these two metabolites. Methods Human liver microsomal incubations, chemical inhibitors, specific antibodies, and different cytochrome P450 expression systems were used. Results K_m and V_max values determined in human liver microsomes were lower for the demethylation (61±21 μm, 159±42 pmol min⁻¹ mg protein⁻¹ mean±s.d.; n=4), than for the N-oxidation of CLZ (308±1.5 μm, 456±167pmol min⁻¹ mg protein⁻¹; n=3). Formation of DCLZ was inhibited by fluvoxamine (53±28% at 10 μm ), triacetyloleandomycin (33±15% at 10 μm ), and ketoconazole (51±28% at 2 μm ) and by antibodies against CYP1A2 and CYP3A4. CLZ-NO formation was inhibited by triacetyloleandomycin (34±16% at 10 μm ) and ketoconazole (51±13% at 2 μm ), and by antibodies against CYP3A4. There was a significant correlation between CYP3A content and DCLZ formation in microsomes from 15 human livers (r=0.67; P=0.04). A high but not significant correlation coefficient was found for CYP3A content and CLZ-NO formation (r=0.59; P=0.09). Using expression systems it was shown that CYP1A2 and CYP3A4 formed DCLZ and CLZ-NO. K_m and V_max values were lower in the CYP1A2 expression system compared to CYP3A4 for both metabolic reactions. Conclusions It is concluded that CYP1A2 and CYP3A4 are involved in the demethylation of CLZ and CYP3A4 in the N-oxidation of CLZ. Close monitoring of CLZ plasma levels is recommended in patients treated at the same time with other drugs affecting these two enzymes.     

Collagen type I and III occur together in hybrid fibrils in the space of Disse of normal rat liver

Collagen type I and procollagen type III were localized at the ultrastructural level on ultrathin frozen sections of rat liver by the protein A-gold technique using affinity-purified primary antibodies. Both collagen type I and procollagen type III were localized on nearly all solitary and bundled fibrils in the space of Disse. Simultaneous localization of collagen type I and procollagen type III by a double-labeling procedure using protein A-gold probes of different sizes unequivocally demonstrated the presence of both collagens in the same fibrils. Measurement of the diameter of large numbers of collagen fibrils in the space of Disse of the rat liver showed a unimodal distribution of the fibril diameters around an average value of 62.4 nm (S.D. = 12.8 nm), and 91% of the collagen bundles contained less than 30 fibrils. Additional measurements on epoxy resin-embedded material of five biopsy specimens of normal human liver showed a comparable unimodal distribution of the fibril diameters around an average value of 57.2 nm (S.D. = 9.6 nm), and 74% of the bundles contained less than 60 fibrils. The latter observation demonstrates that human liver contains broader interstitial collagen bundles than rat liver. From these results, we conclude that the space of Disse of normal rat and human liver contains a uniform population of striated interstitial collagen fibrils. In the rat liver, these fibrils contain both collagen type I and procollagen type III. Therefore the concept that procollagen type III is predominantly localized in small diameter fibrils or bundles, whereas collagen type I is preferentially localized in thick ones, does not hold.     

High-throughput genotyping of thiopurine S-methyltransferase by denaturing HPLC

BACKGROUND: The thiopurine S:-methyltransferase (TPMT) genetic polymorphism has a significant clinical impact on the toxicity of thiopurine drugs, which are used in the treatment of leukemia and as immunosuppressants. To date, 10 mutant alleles are known that are associated with intermediate or low TPMT activity. To facilitate rapid screening of clinically relevant TPMT mutations, we developed a strategy of high-throughput genotyping by applying denaturing HPLC (DHPLC). METHODS: To test the specificity and efficiency of the DHPLC method, 98 DNA samples from a selected population of patients receiving thiopurine therapy or with previous thiopurine withdrawal were analyzed for the most frequent mutant TPMT alleles, *2 and *3A, which contain key mutations in exons 5, 7, and 10 to identify clearly different elution profiles. All fragments were examined by direct sequencing. Additionally, to test the sensitivity of DHPLC analysis, genotyping for the *2 and *3A alleles of all 98 DNA samples was performed by PCR-based methods (PCR-restriction fragment polymorphism analysis and allele-specific PCR). RESULTS: The presence of mutations discriminating for alleles *2, *3A, *3C, and *3D, as well as various silent and intron mutations, were correctly predicted by DHPLC in 100% of the samples as confirmed by direct sequencing. Comparison with PCR-based methods for alleles *2 and *3 produced an agreement of 100% with no false-negative signals. CONCLUSIONS: DHPLC offers a highly sensitive, rapid, and efficient method for genotyping of the relevant TPMT mutations, discriminating at least for alleles *2 and *3, in clinical and laboratory practice. Additionally, DHPLC allows a simultaneous screening for novel genetic variability in the TPMT gene.     

Comprehensive analysis of pyrimidine metabolism in 450 children with unspecific neurological symptoms using high–pressure liquid chromatography–electrospray ionization tandem mass spectrometry

Summary To evaluate the significance of inborn metabolic disorders of the pyrimidine degradation pathway, 450 children with unspecific neurological symptoms were comprehensively studied; 200 healthy children were recruited as controls. Uracil and thymine as well as their degradation products in urine were determined with an improved method based on reversed-phase HPLC coupled with electrospray ionization tandem mass spectrometry and detection by multiple-reaction monitoring using stable-isotope-labelled reference compounds as internal standards. From the results of the control group we established age-related reference ranges of all pyrimidine degradation products. In the patient group, two children with dihydropyrimidine dehydrogenase (DPYD) deficiency were identified; one of these was homozygous for the exon 14-skipping mutation of the DPYD gene. In addition, two patients with high uracil, dihydrouracil and β-ureidopropionate were found to have ornithine transcarbamylase deficiency. In the urine of 9 patients, β-alanine was markedly elevated owing to treatment with vigabatrin, an irreversible inhibitor of GABA transaminase, which interferes with β-alanine breakdown. Four patients had exclusively high levels of β-aminoisobutyrate (β-AIB) due to a low activity of the D-β-AIB-pyruvate aminotransferase, probably without clinical significance. In conclusion, quantitative investigation of pyrimidine metabolites in children with unexplained neurological symptoms, particularly epileptic seizures with or without psychomotor retardation, can be recommended as a helpful tool for diagnosis in clinical practice. Sensitive methods and age-related reference ranges enable the detection of partial enzyme deficiencies. Both authors contributed equally.     

CC531s colon carcinoma cells induce apoptosis in rat hepatic endothelial cells by the Fas/FasL‐mediated pathway

The mechanisms involved in colorectal carcinoma with liver metastasis are not well known. Metastasizing colon carcinoma cells express more FasL than primary colon carcinoma cells and cancer cells induce apoptosis in hepatocytes by the Fas/FasL pathway. Therefore, this study focused on Fas/FasL expression and functionality in rat liver sinusoidal endothelial cells (LSECs) and CC531s colon carcinoma cells in vitro and in vivo. RT‐PCR and immunochemistry revealed Fas and FasL in LSECs and CC531s, respectively. Functionality of Fas was assessed in vitro by incubation with human recombinant FasL (1–100 ng/ml) with or without enhancer. At concentrations of 10 and 100 ng/ml with enhancer, respectively 21% and 44% of endothelial cells showed signs of apoptosis using Hoechst 33342/propidium iodide staining and electron microscopy. In co‐cultures, apoptosis could be detected in endothelial cells neighboring the CC531s and could be inhibited by an antagonistic FasL antibody. Moreover, 18 h after mesenteric injection of CC531s, the sinusoidal endothelium revealed disruption. In conclusion, (i) CC531s cells induce apoptosis in LSECs in vitro by using Fas/FasL; (ii) CC531s cells damage the sinusoidal endothelial lining in vivo; and (iii) this might provide FasL‐positive tumor cells a gateway towards the hepatocytes.