A miniaturized pressure sensor with inherent biofouling protection designed for in vivo applications.

The design, fabrication, and measurement results for a diaphragm-based single crystal silicon sensor element of size 820 µm × 820 µm × 500 µm are presented. The sensor element is designed for in vivo applications with respect to size and measurement range. Moreover, it is optimized for longtime operation in the human body through a built-in protection preventing biofouling on the piezoresistors. The sensitivity is about 20 mV/V for a change from 500 to 1500 mbar absolute pressure. This result is comparable to conventional sized micromachined pressure sensors. The output signal is not found to be influenced by exposure to 60 °C for three hours, a normal temperature load for a typical sterilization process for medical devices (Ethylene Oxide Sterilization). The hysteresis is low; < 0.25% of full scale output signal. The sensor element withstands an overload pressure of 3000 mbar absolute pressure. Observed decrease in the output signal with temperatures and observed nonlinearity can easily be handled by traditional electronic compensation techniques.

[Mutational analysis of VSX-1 in one patient with posterior polymorphous corneal dystrophy and in three families with hereditary Fuchs endothelial dystrophy]. / Mutationsanalyse im VSX1-Gen bei einem Patienten mit posteriorer polymorpher Hornhautdystrophie und drei Familien mit autosomal-dominant vererbter Fuchs'scher Hornhautendotheldystrophie.

BACKGROUND: Fuchs endothelial corneal dystrophy is the most common disease of the corneal endothelium. Besides some sporadic cases, an autosomal dominant inheritance is frequently described. Mutations in the VSX-1 gene are identified as the underlying gene defect for a rarer kind of endothelial dystrophy, posterior polymorphous endothelial dystrophy. We report on mutational analysis of the VSX-1 gene in affected and non-affected family members of three families with autosomal dominant inherited Fuchs endothelial corneal dystrophy and one male patient showing posterior polymorphous endothelial dystrophy. PATIENTS/MATERIALS AND METHODS: From one patient with posterior polymorphous endothelial dystrophy and 10 affected and 15 non-affected family members of three families with autosomal dominant inherited Fuchs endothelial corneal dystrophy DNA was extracted from leukocytes of the peripheral blood and mutational analysis was performed by direct sequencing of the VSX-1 gene. RESULTS: Screening of the VSX-1 gene did not reveal sequence variants in any affected or non-affected individuals from the three families with Fuchs endothelial corneal dystrophy or the patient with posterior polymorphous endothelial dystrophy. CONCLUSIONS: The absence of pathogenic mutations in the VSX-1 gene in affected family members of 3 pedigrees indicates that other genetic factors are involved in the development of familial Fuchs endothelial corneal dystrophy. In addition, VSX-1 seems unlikely to be the crucial gene in our patient with posterior polymorphous endothelial dystrophy.

TGFBI (BIGH3) gene mutations in German families: two novel mutations associated with unique clinical and histopathological findings.

BACKGROUND: To report the clinical, histopathological and immunohistochemical findings of two novel mutations within the TGFBI gene. METHODS: The genotype of 41 affected members of 16 families and nine sporadic cases was investigated by direct sequencing of the TGFBI gene. Clinical, histological and immunohistochemical characteristics of corneal opacification were reported and compared with the coding region changes in the TGFBI gene. RESULTS: A novel mutation Leu509Pro was detected in one family with a geographic pattern-like clinical phenotype. Histopathologically we found amyloid together with non-amyloid deposits and immunohistochemical staining of Keratoepithelin (KE) KE2 and KE15 antibodies. In two families and one sporadic case the novel mutation Gly623Arg with a late-onset, map-like corneal dystrophy was identified. Here amyloid and immunohistochemical staining of only KE2 antibodies occurred. Further, five already known mutations are reported: Arg124Cys Arg555Trp Arg124His His626Arg, Ala546Asp in 13 families and five sporadic cases of German origin. The underlying gene defect within the TBFBI gene was not identified in any of the four probands with Thiel-Behnke corneal dystrophy. CONCLUSIONS: The two novel mutations within the TGFBI gene add another two phenotypes with atypical immunohistochemical and histopathological features to those so far reported.

[Idiopathic orbital inflammatory syndrome (orbital pseudotumors): diagnosis and therapy]. / Idiopathische entzündliche intraorbitale Fibrosklerose: Diagnostik und therapeutische Möglichkeiten.

BACKGROUND: Pseudotumors of the orbit comprise a group of idiopathic inflammatory processes and are, except for endocrine orbitopathy, the most common reason for exophthalmos in adults. Orbital pseudotumors, also called idiopathic orbital inflammatory syndrome (IOIS), can be determined from orbital involvement in systemic fibrosing diseases. Finding the correct diagnosis can be challenging. Due to the topographic relations of the orbit to neighbouring structures, a multidisciplinary cooperation is highly recommended. CASE REPORT: We report a case of a 42-year-old woman with unilateral exophthalmos. Additionally we found impaired motility of the affected bulbus, ptosis and reduction of visual acuity. Orbital MR imaging demonstrated dense fibrotic masses filling the whole orbita including the extraocular muscles as well as the optic nerve. Tissue specimens were extracted while performing orbital decompression via a lateral orbitotomy. Histological examination revealed a lymphatic infiltration and fibrotically destroyed tissue containing the lacrimal gland. After surgical decompression, oral steroid therapy and immunotherapy, a recovery of the visual loss could be seen. CONCLUSIONS: Intraorbital fibrosclerosing pseudotumors often require a difficult long-term treatment. Therapeutic options are steroid therapy, immunotherapy, radiotherapy and surgery. The diagnostic steps include blood tests, ultrasound, CT and/or MRI as well as histological differentiation. Solid tumors and orbital involvement in diseases of the hematopoetic system have to be excluded. Since intraorbital fibrosis can be accompanied by manifestations in various other organs, a complete investigation of the body and thorough follow up are crucial.

The complete genome of the crenarchaeon Sulfolobus solfataricus P2.

The genome of the crenarchaeon Sulfolobus solfataricus P2 contains 2,992,245 bp on a single chromosome and encodes 2,977 proteins and many RNAs. One-third of the encoded proteins have no detectable homologs in other sequenced genomes. Moreover, 40% appear to be archaeal-specific, and only 12% and 2.3% are shared exclusively with bacteria and eukarya, respectively. The genome shows a high level of plasticity with 200 diverse insertion sequence elements, many putative nonautonomous mobile elements, and evidence of integrase-mediated insertion events. There are also long clusters of regularly spaced tandem repeats. Different transfer systems are used for the uptake of inorganic and organic solutes, and a wealth of intracellular and extracellular proteases, sugar, and sulfur metabolizing enzymes are encoded, as well as enzymes of the central metabolic pathways and motility proteins. The major metabolic electron carrier is not NADH as in bacteria and eukarya but probably ferredoxin. The essential components required for DNA replication, DNA repair and recombination, the cell cycle, transcriptional initiation and translation, but not DNA folding, show a strong eukaryal character with many archaeal-specific features. The results illustrate major differences between crenarchaea and euryarchaea, especially for their DNA replication mechanism and cell cycle processes and their translational apparatus.

Two different and highly organized mechanisms of translation initiation in the archaeon Sulfolobus solfataricus.

The translational starts of 144 Sulfolobus solfataricus genes have been determined by database comparison. Half the genes lie inside operons and the other half are at the start of an operon or single genes. A Shine-Dalgarno sequence is found upstream of the genes inside operons, but not for the first gene in an operon or isolated genes; this indicates that two different mechanisms are used for translation initiation in S. solfataricus. A box A transcriptional signal is found for the genes starting an operon or isolated genes, but not for the genes inside an operon. The box A signal is located about 27 nt upstream of the start codon, which implies that little or no upstream sequence is available for translation initiation for this group of genes. This finding is discussed.

Completing the sequence of the Sulfolobus solfataricus P2 genome.

The Sulfolobus solfataricus P2 genome collaborators are poised to sequence the entire 3-Mbp genome of this crenarchaeote archaeon. About 80% of the genome has been sequenced to date, with the rest of the sequence being assembled fast. In this publication we introduce the genomic sequencing and automated analysis strategy and present intial data derived from the sequence analysis. After an overview of the general sequence features, metabolic pathway studies are explained, using sugar metabolism as an example. The paper closes with an overview of repetitive elements in S. solfataricus.

The role of Glu87 and Trp89 in the lid of Humicola lanuginosa lipase.

The importance of Glu87 and Trp89 in the lid of Humicola lanuginosa lipase for the hydrolytic activity at the water/lipid interface was investigated by site-directed mutagenesis. It was found that the effect on the hydrolytic activity upon the replacement of Trp89 with Phe, Leu, Gly or Glu was substrate dependent. The Trp89 mutants displayed an altered chain length specificity towards triglycerides, with a higher relative activity towards triacetin and trioctanoin compared with tributyrin. Trp89 was shown to be less important in the hydrolysis of vinyl esters compared with ethyl esters and triglycerides. An exclusive effect on the acylation reaction rate by the mutation of Trp89 was consistent with the data. It is suggested that Trp89 is important in the process of binding the acyl chain of the substrate into the active site for optimal acylation reaction rate. The Trp89Phe mutation resulted in an increased hydrolytic activity towards 2-alkylalkanoic acid esters. This is suggested to be due to reduction of unfavourable van der Waals contacts between Trp89 and the 2-substituent of the substrate. Thus, in contrast to natural substrates, Trp89 has a negative impact on the catalytic efficiency when substrates with bulky acyl chains are used. In contrast to the Trp89 mutations, the effect on the hydrolytic activity of the Glu87Ala mutation was almost substrate independent, 35-70% activity of wild-type lipase. A reduction of both the acylation and deacylation reaction was consistent with the data.

Conservative versus radical surgery for tubal pregnancy. A review.

OBJECTIVE: The aim of this work was to analyse the fertility prognosis after conservative or radical surgery for tubal pregnancy. DATA SOURCES: Index Medicus was searched for all attainable literature on the subject. METHODS OF STUDY SELECTION: A total of 40 scientific publications through the latest 40 years were selected. For fulfilling the selection criterias the study design should appear clearly. Furthermore the rate of women obtaining intrauterine pregnancy and the rate of repeat ectopic pregnancy following radical or conservative tubal surgery was to be compared using 95% confidence limits. The results from each report were compared in four groups according to study design i.e. retrospective non-comparing materials, retrospective comparing studies, prospective selected treatment series or prospective randomized comparing investigations. DATA EXTRACTION AND SYNTHESIS: Pooling the results from the retrospective noncomparing materials revealed that there was no significant difference in intrauterine pregnancy rates, i.e. 46% following conservative tubal surgery and 44% after radical surgery. The repeat ectopic pregnancy rate was 10% following conservative surgery and 15% after radical surgery. In the group of restrospective comparing studies only one of 15 materials could document a significant better intrauterine pregnancy rate after conservative tubal surgery than following radical treatment for tubal pregnancy. There were no differences either in this group in repeat ectopic pregnancy rates. Prospective investigations were almost exclusively represented by selected conservative treatment series. In these series the average intrauterine pregnancy rate was 57% and the repeat ectopic pregnancy rate was 13%. CONCLUSIONS: In studies on fertility after radical or conservative surgical treatment for tubal pregnancy no significant difference in intrauterine pregnancy rates or repeat ectopic pregnancy rates were found. Prospective selected treatment series demonstrated higher intrauterine pregnancy rates than retrospective studies. The repeat ectopic pregnancy rate was not raised in prospective series. No prospective randomised trial was found.

Isolation and characterisation of genes for sulphate activation and reduction in Aspergillus nidulans: implications for evolution of an allosteric control region by gene duplication.

A region of the Aspergillus nidulans genome carrying the sA and sC genes, encoding PAPS reductase and ATP sulphurylase, respectively, was isolated by transformation of an sA mutant with a cosmid library. The genes were subcloned and their functions confirmed by retransformation and complementation of A. nidulans strains carrying sA and sC mutations. The physical distance of 2 kb between the genes corresponds to a genetic distance of 1 cM. While the deduced amino acid sequence of the sA gene product shows homology with the equivalent MET16 gene product of Saccharomyces cerevisiae, the sC gene product resembles the equivalent MET3 yeast gene product at the N-terminal end, but differs markedly from it at the C-terminal end, showing homology to the APS kinases of several microorganisms. It is proposed that this C-terminal region does not encode a functional APS kinase, but is responsible for allosteric regulation by PAPS of the sulphate assimilation pathway in A. nidulans, and that the ATP sulphurylase encoding-gene (sC) of filamentous ascomycetes may have evolved from a bifunctional gene similar to the nodQ gene of Rhizobium meliloti.

Probing a functional role of Glu87 and Trp89 in the lid of Humicola lanuginosa lipase through transesterification reactions in organic solvent.

To reveal the functional role of Glu87 and Trp89 in the lid of Humicola lanuginosa lipase, site-directed mutagenesis at Glu87 and Trp89 was carried out. The catalytic performance of wild-type and mutated lipases was studied in transesterification reactions in cyclohexane at a controlled water activity. Two different acyl donors were used in the investigation: tributyrin, a natural substrate for a lipase, and vinyl butyrate, an activated ester suitable for fast and efficient lipase-catalyzed transformations in preparative organic synthesis. As acyl acceptor 1-heptanol was used. The Glu87Ala mutation decreased the Vmax,app value with tributyrin and vinyl butyrate by a factor of 1.5 and 2, respectively. The Km,app for tributyrin was not affected by the Glu87Ala mutation, but the Km,app for vinyl butyrate increased twofold compared to the wild-type lipase. Changing Trp89 into a Phe residue afforded an enzyme with a 2.7- and 2-fold decreased Vmax,app with the substrates tributyrin and vinyl butyrate, respectively, compared to the wild-type lipase. No significant effects on the Km,app values for tributyrin or vinyl butyrate were seen as a result of the Trp89Phe mutation. However, the introduction of a Glu residue at position 89 in the lid increased the Km,app for tributyrin and vinyl butyrate by a factor of > 5 and 2, respectively. The Trp89Glu mutated lipase could not be saturated with tributyrin within the experimental conditions (0-680 mM) studied here. With vinyl butyrate as a substrate the Vmax,app was only 6% of that obtained with wild-type enzyme.

Trp89 in the lid of Humicola lanuginosa lipase is important for efficient hydrolysis of tributyrin.

To determine whether Trp89 located in the lid of the lipase (EC 3.1.1.3) from Humicola lanuginosa is important for the catalytic property of the enzyme, site-directed mutagenesis at Trp89 was carried out. The kinetic properties of wild type and mutated enzymes were studied with tributyrin as substrate. Lipase variants in which Trp89 was changed to Phe, Leu, Gly or Glu all showed less than 14% of the activity compared to that of the wild type lipase. The Trp89Glu mutant was the least active with only 1% of the activity seen with the wild type enzyme. All Trp mutants had the same binding affinity to the tributyrin substrate interface as did the wild type enzyme. Wild type lipase showed saturation kinetics against tributyrin when activities were measured with mixed emulsions containing different proportions of tributyrin and the nonionic alkyl polyoxyethylene ether surfactant, Triton DF-16. Wild type enzyme showed a Vmax = 6000 +/- 300 mmol.min-1.g-1 and an apparent Km = 16 +/- 2% (vol/vol) for tributyrin in Triton DF-16, while the mutants did not show saturation kinetics in an identical assay. The apparent Km for tributyrin in Triton DF-16 was increased as the result of replacing Trp89 with other residues (Phe, Leu, Gly or Glu). The activities of all mutants were more sensitive to the presence of Triton DF-16 in the tributyrin substrate than was wild type lipase. The activity of the Trp89Glu mutant was decreased to 50% in the presence of 2 vol% Triton DF-16 compared to the activity seen with pure tributyrin as substrate.(ABSTRACT TRUNCATED AT 250 WORDS)

An extra-uterine pregnancy in an ectopic fallopian tube.

A case report of an ectopic pregnancy in a non-communicating, undescended fallopian tube. A rare embryological development-failure is described, consisting of a right hemiuterus, a remotely located noncommunicating left fallopian tube and undescended left ovary associated with agenesis of the left kidney. A noncommunicating fallopian tube should be removed when recognized to prevent ectopic pregnancy.

Lipases from Rhizomucor miehei and Humicola lanuginosa: modification of the lid covering the active site alters enantioselectivity.

The homologous lipases from Rhizomucor miehei and Humicola lanuginosa showed approximately the same enantioselectivity when 2-methyldecanoic acid esters were used as substrates. Both lipases preferentially hydrolyzed the S-enantiomer of 1-heptyl 2-methyldecanoate (R. miehei: ES = 8.5; H. lanuginosa: ES = 10.5), but the R-enantiomer of phenyl 2-methyldecanoate (ER = 2.9). Chemical arginine specific modification of the R. miehei lipase with 1,2-cyclohexanedione resulted in a decreased enantioselectivity (ER = 2.0), only when the phenyl ester was used as a substrate. In contrast, treatment with phenylglyoxal showed a decreased enantioselectivity (ES = 2.5) only when the heptyl ester was used as a substrate. The presence of guanidine, an arginine side chain analog, decreased the enantioselectivity with the heptyl ester (ES = 1.9) and increased the enantioselectivity with the aromatic ester (ER = 4.4) as substrates. The mutation, Glu 87 Ala, in the lid of the H. lanuginosa lipase, which might decrease the electrostatic stabilization of the open-lid conformation of the lipase, resulted in 47% activity compared to the native lipase, in a tributyrin assay. The Glu 87 Ala mutant showed an increased enantioselectivity with the heptyl ester (ES = 17.4) and a decreased enantioselectivity with the phenyl ester (ER = 2.5) as substrates, compared to native lipase. The enantioselectivities of both lipases in the esterification of 2-methyldecanoic acid with 1-heptanol were unaffected by the lid modifications.

[Fetal-pelvic disproportion]. / Disproportio foetopelvina.

The files of 186 primiparous women with the diagnosis of fetal-pelvic disproportion were studied in order to depict maternal and fetal parameters determining the course of delivery. Three groups were compared. One group of women who delivered vaginally and two groups, that delivered by caesarean section, one with a dilated orifice and one with a non-dilated orifice. The maternal age, height and pelvic capaciousness as found by clinical examination were registered together with cardiotocography, birth weight, labour augmentation, instrumental delivery, fetal presentation, gestation age and the conjugata vera (measured at caesarian section). It was found that the maternal age and the gestational age were lower in the vaginal delivery group compared to the two caesarean section groups. There was no difference between all three groups with respect to the other parameters. On this basis it was concluded, that it was not possible to identify fetal-pelvic disproportion that would result in caesarean section in primiparous women.

Stereomicroscopic demonstration of chorionic villi: differentiation between miscarriage and ectopic pregnancy.

OBJECTIVE: To evaluate direct peroperative stereomicroscopic examination of endometrial curettings in the differentiation between miscarriage and ectopic pregnancy. DESIGN: A prospective consecutive controlled study. SETTING: Odense University Hospital, Odense, Denmark. SUBJECTS: One hundred and fifty-nine women with vaginal bleeding and/or abdominal pain in early pregnancy and no fetus seen on ultrasound. MAIN OUTCOME MEASURES: The results of stereomicroscopy were compared with the histological diagnoses. RESULTS: There was 90% (95% CI, 84-94%) agreement between the stereomicroscopic and histological examinations. Fifteen per cent had an ectopic pregnancy. The sensitivity of stereomicroscopy as a marker of ectopic pregnancy was 92% (73-99%), the specificity 94% (88-97%). The predictive value of a positive test was 73% (54-88%) and that of a negative test 98% (94-100%). CONCLUSION: Direct peroperative stereomicroscopy of endometrial curettings is a valuable diagnostic tool for immediate differentiation between miscarriage and ectopic pregnancy.