Comparison of MR imaging findings between extraligamentous and subligamentous disk herniations in the lumbar spine.

BACKGROUND AND PURPOSE: The method of treating an HIVD in the lumbar spine may depend on the integrity of the PLL. The purpose of this study was to analyze and compare the MR imaging findings of extraligamentous and subligamentous HIVDs in the lumbar spine. MATERIAL AND METHODS: One hundred seventeen patients (M/F = 71:46; mean age, 47 years; age range, 15-79 years) underwent lumbar spine MR imaging and disk surgery (extraligamentous/subligamentous = 66:51) from May 2003 to November 2006. Two radiologists in consensus retrospectively reviewed all MR images, focusing on 10 criteria. RESULTS: The following 5 criteria are suggestive of extraligamentous HIVD in the lumbar spine: 1) spinal canal compromised for more than half its dimension, 2) internal signal difference in the HIVD, 3) an ill-defined margin of the HIVD, 4) disruption of the continuous low-signal-intensity line covering the HIVD, and 5) the presence of an internal dark line in the HIVD (P < .05). When we combined these 5 MR imaging criteria, the sensitivity, specificity, accuracy, and odds ratio were 77.3%, 74.5%, 76.1%, and 9.93 (P < .0001). CONCLUSIONS: Our proposed 5 MR imaging criteria will be helpful in differentiating extraligamentous and subligamentous HIVDs in the lumbar spine.

Characteristic radiographic features of the central ray in Apert syndrome.

We reviewed retrospectively seven patients with Apert acrosyndactyly and measured the size of the capitate ossification centre relative to that of the hamate and determined the relative position of the middle finger metacarpal relative to the ring finger metacarpal. We then compared those parameters in 197 normal children. In all patients, the middle finger metacarpal bone had migrated proximally relative to the ring finger metacarpal and the size of the capitate ossification centre was smaller than that of the hamate. After surgical release of the middle finger, relative proximal migration of the middle finger metacarpal was partially relieved and catch-up growth of the capitate was observed within several months. As fusion of the distal phalanges creates a diamond-shaped configuration, bone growth is markedly impaired in the middle finger ray. Therefore, early separation of the middle finger may be as important as early separation of the border digits.

Intrasac pressure changes and vascular remodeling after endovascular repair of abdominal aortic aneurysms: review and biomechanical model simulation.

In this paper, we review existing clinical research data on post-endovascular repair (EVAR) intrasac pressure and relation with abdominal aortic aneurysm (AAA) size changes. Based on the review, we hypothesize that intrasac pressure has a significant impact on post-EVAR AAA size changes, and post-EVAR remodeling depends also on how the pressure has changed over a period of time. The previously developed model of an AAA based on a constrained mixture approach is extended to include vascular adaptation after EVAR using an idealized geometry. Computational simulation shows that the same mechanism of collagen stress-mediated remodeling in AAA expansion induces the aneurysm wall to shrink in a reduced sac-pressure after post-EVAR. Computational simulation suggests that the intrasac pressure of 60 mm Hg is a critical value. At this value, the AAA remains stable, while values above cause the AAA to expand and values below cause the AAA to shrink. There are, however, variations between individuals due to different cellular sensitivities in stress-mediated adaptation. Computer simulation also indicates that an initial decrease in intrasac pressure helps the AAA shrink even if the pressure increases after some time. The presented study suggests that biomechanics has a major effect on initial adaptation after EVAR and also illustrates the utility of a computational model of vascular growth and remodeling in predicting diameter changes during the progression and after the treatment of AAAs.

The development of rapid real-time PCR detection system for Vibrio parahaemolyticus in raw oyster.

AIMS: To develop a new rapid real-time polymerase chain reaction (PCR) based detection system for Vibrio parahaemolyticus (V. parahaemolyticus) applicable to raw oyster samples. METHODS AND RESULTS: V. parahaemolyticus cells were artificially inoculated to oysters. Samples were homogenized in 100 ml of sterile saline water and serially diluted to 1.5 CFU ml(-1) level. One millilitre of diluents was centrifuged and the pellet was resuspended with 100 microl of de-ionized water. DNA was extracted by boiling for 20 min, and 0.5 microl was used as a template for PCR reaction. Real-time PCR was performed with TMC-1000 system (1 microl PCR system). The detection system was found to achieve detection limit of 1.5 CFU g(-1) for V. parahaemolyticus. Furthermore, the specificities of these assay systems were confirmed with more than 20 bacterial strains, including various Vibrio species. CONCLUSIONS: Rapid and sensitive food-borne pathogen detection techniques for V. parahaemolyticus is important to the food industry and consumers. The direct detection of V. parahaemolyticus from food is possible with micro real-time PCR system. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that oyster samples can be tested for V. parahaemolyticus with a rapid, specific and simple procedure.

[The role of perineal ultrasound compared to lateral cysturethrogram in urogynecological evaluations]. / Die Perinealsonographie im Vergleich zum lateralen Zysturethrogramm bei der Inkontinenzdiagnostik der Frau.

PURPOSE: The aim of this study was to evaluate prospectively whether perineal ultrasound is comparable to the lateral cysturethrogram in the evaluation of incontinent women. PATIENTS AND METHODS: Following urodynamic investigations, a lateral cysturethrogram and perineal ultrasound (5 MHz probe, bladder filling 300 mL) were performed in 98 incontinent women. In women with detrusor overactivity and consecutively reduced bladder capacity, ultrasound was performed at maximum capacity. To evaluate differences between perineal ultrasound and the cysturethrogram, the difference between bladder neck and lower border of symphysis and the retrovesicle angle beta were determined at rest and during the Valsalva manoeuvre. RESULTS: Using perineal ultrasound, the differences between bladder neck and symphysis could be determined at rest and during the Valsalva manoeuvre in all patients. The determination of the retrovesical angle beta was possible in all patients at rest and in 89 of the 98 women during the Valsalva manoeuvre. The lateral cysturethrogram enabled the determination of difference between bladder neck and symphysis and the retrovesicle angle beta at rest in 81 of 98 women. During the Valsalva manoeuvre, the difference between bladder neck and symphysis and retrovesicle angle beta could be determined in 72 of the 98 women. In the 26 remaining women, the determination was impossible due to severe adiposity or cystoceles of the second or third degree. CONCLUSIONS: Perineal ultrasound provides comparable data to the lateral cysturethrogram. In patients with adiposity, perineal ultrasound seems to be superior. Within the routine evaluation of women suffering from incontinence, the lateral cysturethrogram can be replaced by perineal ultrasound without any limitations of the diagnostic value.

[New techniques for surgical treatment of postoperative male stress incontinence]. / Neue Verfahren zur operativen Therapie der postoperativen Belastungsinkontinenz beim Mann.

The impact of minimally invasive techniques for the treatment of postoperative male incontinence has significantly improved in recent years. These techniques are based on the continuous increase in urethral resistance. This resistance can be readjusted with balloons placed paraurethrally or with readjustable suburethral slings. The success rates depend on the preoperative degree of incontinence. With bulking agents that are transurethrally injected into the submucosa of the sphincter, continence rates between 12 and 90% can be seen. Severe complications are rare. The impact of the studies is often limited due to a short follow-up. After implantation of adjustable balloons that are placed paraurethrally close to the bladder neck, continence rates up to 70% can be seen. The overall improvement of incontinence is observed in up to 90% of the treated patients. Complications such as balloon migration or mechanical disorders can cause operative revision. Suburethral sling systems are available as bone-anchored slings or as readjustable slings. Continence can be seen in up to 90% of the patients postoperatively. Severe complications such as sling erosion or sling infection are rare. In cases of mild and moderate incontinence, these minimally invasive techniques are good alternatives to the fascial sling or alloplastic sphincter implantation. To improve the evaluation and to compare these techniques with the conventional methods, further investigations with a longer follow-up are necessary.

Hans Christian Jacobaeus: Inventor of human laparoscopy and thoracoscopy.

Hans Christian Jacobaeus performed the first clinical laparoscopic surgery in Stockholm. This pioneering procedure was based on the animal experiments of Georg Kelling (1866-1945), a German physician from Dresden, who performed the first laparoscopic intervention in 1901 using a Nitz cystoscope in a dog. In 1910, Jacobaeus published his initial experiences with laparoscopic surgery in the Münchner Medizinischen Wochenschrift under the title "The Possibilities for Performing Cystoscopy in Examinations of Serous Cavities." He used this technique for diagnostic purposes in undefined abdominal complaints and functional impairment. Jacobaeus was the first who pointed out the possibility of injuring organs, especially the intestines, by inserting the trocar. In 1910, Jacobaeus recognized the immense diagnostic and therapeutic possibilities of laparoscopic surgery, as well as its difficulties and limits. He also was the first to realize the need for initial endoscopic training in animals and corpses. He promoted the development of special laparoscopic instruments to optimize and simplify the procedure.

Assisted procreation in cases of hepatitis B, hepatitis C or human immunodeficiency virus infection of the male partner.

Guidelines for assisted procreation impose a special responsibility upon physicians for the health of the expected child because of their active role in inducing pregnancy. Therefore, careful clinical evaluation of both partners has to precede every application of these methods. Risks for the mother's health or the development of the child count as a relative contraindication for a treatment. To balance these relative contraindications, the existing risk factors have to be recognized through screening examination. If a chronic infection occurs in the male partner, prevention for the female partner is theoretically possible by using a condom. As this inhibits a pregnancy, at least in cases of human immunodeficiency virus and hepatitis C virus infections, realization of a pregnancy requires assisted procreation. The main question in these cases is whether infectious particles can be eliminated by sperm processing to ensure the safe treatment of the healthy female partner.

Distraction osteogenesis for segmental bone defect. Physeal change after acute bone shortening followed by gradual lengthening in a rabbit tibia model.

We investigated physeal change after acute bone shortening followed by distraction osteogenesis in a rabbit tibia model. Distraction osteogenesis was performed on acutely shortened right rabbit tibia after removing a diaphyseal segment until distraction reached the length of the excised segment. Sixty animals were divided into three groups (n=20) according to the amount of shortening (20, 30, and 40% of original length) and radiographic, immunohistochemical, and histomorphometric analyses were performed to evaluate the physeal activity. The results showed no differences between groups and all animals could recover the original length without growth retardation. There was no notable venous congestion or lymphatic kinking associated with acute shortening. These results suggest that distraction osteogenesis at the primary site of shortening may have little adverse effects on the growth plate, when the shortening can provide sufficient soft tissue without vascular compromise and the amount of lengthening does not exceed that of shortening.

Production of fructo-oligosaccharides from molasses by Aureobasidium pullulans cells.

Three different strains of Aureobasidium pullulans were grown in batch cultures to compare their abilities for the production of fructo-oligosaccharides. Specific intracellular enzyme activity was the highest with strain KCCM 12017 and enzyme production was closely coupled to growth. Using A. pullulans cells, 166 g/l fructo-oligosaccharides was produced from 360 g/l molasses sugar as sucrose equivalent at 55 degrees C and pH 5.5 after 24 h incubation.

Spine needle biopsy simulator using visual and force feedback.

OBJECTIVE: Biopsy with an inserted needle is an important procedure for lesion detection in the spine, but is difficult to perform due to the presence of many critical organs near the spine. This article presents a spine needle biopsy simulator, based on visual and force feedback, which can be used to plan the optimal path of a needle and to practice the procedure without risk. MATERIALS AND METHODS: The simulator is composed of a 3D human model, a visual-feedback component, a force-feedback component, and an evaluation module. The human model is based on 3D CT data. The visual-feedback component provides an oblique section, multiplanar reformatting images, and a volume-rendered image. Of these, the oblique section display is very useful for planning a 3D path for the needle. During simulation, the force-feedback component generates and provides realistic forces acting on the biopsy needle in real time by synchronizing them to visual feedback. After each simulation, the evaluation module provides a performance analysis for the trainee. RESULTS: For an XCT abdomen volume data set of 256 x 256 x 256, the update rate of image rendering due to needle movement is over 25 Hz, with a force-feedback rate of 1 kHz. This performance proved to be good enough for the trainee to learn the relationship between visual and force feedback. CONCLUSIONS: The simulator is useful for the planning of and training in complicated 3D spine needle biopsy procedures. It may be used as an educational tool for beginners, a practice tool to increase expertise, or a test bed for new procedures.

Expression in Escherichia coli of the thermostable inorganic pyrophosphatase from the Aquifex aeolicus and purification and characterization of the recombinant enzyme.

The gene encoding the inorganic pyrophosphatase from a hyperthermophilic bacterium, Aquifex aeolicus (Aae), was amplified by PCR. Then, the gene was overexpressed in Escherichia coli using a pJR-based expression plasmid, pAIPD. The recombinant Aae pyrophosphatase was purified 16.2-fold with a 53.4% yield and a specific activity of 34 U/mg protein by a combination of heating (to denature E. coli proteins) and two steps of DEAE-Sephacel column chromatography (nonabsorbed enzyme at pH 7.3 and absorbed enzyme at pH 8.0). This enzyme has an approximate molecular mass of 105,000 Da and consists of four subunits, each with a molecular mass of 24,500 Da. The enzyme shows the optimal activity in the pH range 7.5-8.0. The enzyme was stable at 80-95 degrees C. A divalent cation was absolutely required for the enzyme activity, Mg(2+) being most effective.

Cloning, heterologous expression, and functional characterization of 5-epi-aristolochene-1,3-dihydroxylase from tobacco (Nicotiana tabacum).

Capsidiol is a bicyclic, dihydroxylated sesquiterpene produced by several solanaceous species in response to a variety of environmental stimuli. It is the primary antimicrobial compound produced by Nicotiana tabacum in response to fungal elicitation, and it is formed via the isoprenoid pathway from 5-epi-aristolochene. Much of the biosynthetic pathway for the formation of this compound has been elucidated, except for the enzyme(s) responsible for the conversion of 5-epi-aristolochene to its dihydroxylated form, capsidiol. Biochemical evidence from previous studies with N. tabacum (Whitehead, I. M., Threlfall, D. R., and Ewing, D. F., 1989, Phytochemistry 28, 775-779) and Capsicum annuum Hoshino, T., Yamaura, T., Imaishi, H., Chida, M., Yoshizawa, Y., Higashi, K., Ohkawa, H., Mizutani, J., 1995, Phytochemistry 38, 609-613. suggested that the oxidation of 5-epi-aristolochene to capsidiol was mediated by at least one elicitor-inducible cytochrome P450 hydroxylase. In extending these observations, we developed an in vivo assay for 5-epi-aristolochene hydroxylase activity and used it to demonstrate a dose-dependent inhibition of activity by ancymidol and ketoconazole, two well characterized inhibitors of cytochrome P450 enzymes. Using degenerate oligonucleotide primers designed to the well conserved domains found within most P450 enzymes, including the heme binding domain, cDNA fragments representing four distinct P450 families (CYP71, CYP73, CYP82, and CYP92) were amplified from a cDNA library prepared against mRNA from elicitor-treated cells using PCR. The PCR fragments were subsequently used to isolate full-length cDNAs (CYP71D20 and D21, CYP73A27 and A28, CYP82E1 and CYP92A5), and these in turn were used to demonstrate that the corresponding mRNAs were all induced in elicitor-treated cells, albeit with different induction patterns. Representative, full-length cDNAs for each of the P450s were engineered into a yeast expression system, and the recombinant yeast assessed for functional expression of P450 protein by measuring the CO difference spectra of the yeast microsomes. Only microsomal preparations from yeast expressing the CYP71D20 and CYP92A5 cDNAs exhibited significant CO difference absorbance spectra at 450 nm and were thus tested for their ability to hydroxylate 5-epi-aristolochene and 1-deoxycapsidiol, a putative mono-hydroxylated intermediate in capsidiol biosynthesis. Interestingly, the CYP71D20-encoded enzyme activity was capable of converting both 5-epi-aristolochene and 1-deoxycapsidiol to capsidiol in vitro, consistent with the notion that this P450 enzyme catalyzes both hydroxylations of its hydrocarbon substrate.

Purification and properties of Aquifex aeolicus DNA polymerase expressed in Escherichia coli.

The gene encoding Aquifex aeolicus (Aae) DNA polymerase was expressed under the control of the trp promoter on a high-copy plasmid, pTRPNS, in Escherichia coli. The expressed enzyme was purified 11-fold with a 13.8% yield and a specific activity of 2268.3 U mg(-1). The optimum pH of the enzyme was 6.8-7.2. The optimal concentrations of KCl and Mg(2+) were 20-30 mM and 4-5 mM, respectively. Aae DNA polymerase contained a double-strand-dependent 3'-->5' proofreading exonuclease activity but lacked any detectable 5'-->3' exonuclease activity.

Molecular characterization of beta-tubulin gene from Pleurotus sajor-caju.

A beta-tubulin gene (TUB1) from the basidiomycete Pleurotus sajor-caju was sequenced. TUB1 encodes a 446-amino-acid protein. The coding region is interrupted by 9 introns, all of which had a 5'-GTRNGT... YAG-3' sequence at the boundaries. Locations of the introns in TUB1 were common between the beta-tubulin genes of other basidiomycetes, but not with animals, ascomycetes, or plants. This suggests that the introns were inserted independently into the beta-tubulin gene after these divisions had diverged.

Heterotopic pancreas in the stomach: CT findings.

PURPOSE: To describe the computed tomographic (CT) findings of heterotopic pancreas in the stomach. MATERIALS AND METHODS: CT findings in 12 patients with heterotopic pancreas in the stomach were reviewed. Surgical resection (n = 11) or endoscopic excision (n = 1) was performed in cases of symptomatic heterotopic pancreas (n = 4), suspected submucosal tumors (n = 7), and gastric carcinoma (n = 1). Seven patients underwent helical CT with water as an oral contrast agent; five underwent nonhelical CT with water-soluble contrast material. RESULTS: Nine heterotopic pancreata were in the antrum and one each was in the body, fundus, and perigastric fat. Seven lesions were on the greater curvature aspect; five, on the lesser curvature aspect. Common CT findings were well-defined oval or round masses with smooth or serrated margins in the gastric antral wall. Four of the seven lesions in which helical CT was performed enhanced similarly to normal pancreas. Preoperatively, CT depicted 11 of the 12 lesions, but CT findings were interpreted correctly as heterotopic pancreas in only two; the remaining 10 were misinterpreted as other lesions. Atypical findings were cystic dilatation of heterotopic pancreatic duct in two, unusual location in the fundus or perigastric fat in two, and malignant transformation in one. CONCLUSION: CT findings of heterotopic pancreas in the stomach appear to be nonspecific for diagnosis, except for location.

Changes in blood flow during one stage lengthening of bone: an experimental study in rats.

Distraction osteogenesis is a well-accepted method of bone lengthening. Its disadvantages, however, are that it requires an external fixator and takes a long time. One-stage lengthening therefore offers certain advantages. A first point of reference for the safe limits of this procedure might be the changes of blood flow, and this is also the crucial factor in deciding on the appropriate method of lengthening, particularly where the hand or foot is involved. Using a laser Doppler flowmeter we measured blood flow in the dorsum of the foot after using bilateral minimonofixators to lengthen the tibias of 15 Sprague-Dawley rats. They were lengthened in four stages: stage 0 (before lengthening); stage I--12.5%; stage II--25%; and stage III--31.25% of lengthening. The blood flow during stage I decreased to 79% compared to that of stage 0; 16% during stage II; and 1% during stage III. This study suggests that the maximal permissible extent of lengthening might be less than a quarter according to the blood flow as suggested by this animal model.

Crystal structure of NAD(+)-dependent DNA ligase: modular architecture and functional implications.

DNA ligases catalyze the crucial step of joining the breaks in duplex DNA during DNA replication, repair and recombination, utilizing either ATP or NAD(+) as a cofactor. Despite the difference in cofactor specificity and limited overall sequence similarity, the two classes of DNA ligase share basically the same catalytic mechanism. In this study, the crystal structure of an NAD(+)-dependent DNA ligase from Thermus filiformis, a 667 residue multidomain protein, has been determined by the multiwavelength anomalous diffraction (MAD) method. It reveals highly modular architecture and a unique circular arrangement of its four distinct domains. It also provides clues for protein flexibility and DNA-binding sites. A model for the multidomain ligase action involving large conformational changes is proposed.

Crystallization and preliminary x-ray crystallographic analysis of NAD+-dependent DNA ligase from Thermus filiformis.

A highly thermostable DNA ligase from Thermus filiformis has been crystallized at room temperature using methoxypolyethylene glycol 5000 as a precipitant. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 90.63, b = 117.80, c = 98. 65 A, beta = 115.56 degrees. Two molecules of DNA ligase are present in the asymmetric unit, giving a crystal volume per protein mass (V(m)) of 3.1 A(3) Da(-1) and a solvent content of 61%. A native data set extending to 3.0 A resolution has been collected at 100 K using synchrotron X-rays.

Isolation and transformation of uracil auxotrophs of the edible basidiomycete Pleurotus ostreatus.

Uracil auxotrophs of Pleurotus ostreatus were isolated using the selectable marker, resistance to 5'-fluoro-orotic acid (5'-FOA). Two of the nine uracil auxotrophs obtained were transformed to prototrophy using plasmid pTRura 3-2 that contains the orotidine monophosphate decarboxylase (ura3) gene from Trichoderma reesei. Southern blot analyses of the transformants showed that the transforming DNA had integrated into the genome of the protoplasts. Using 2 x 10(7) protoplasts, this system gave a transformation efficiency of about 30 transformants per microg of DNA. Normal fruiting bodies were induced in the transformants by crossing them with wild-type monokaryons, and the basidiospores collected from these fruiting bodies showed a biased segregation rate to prototrophy. These results indicate the integrated DNA was stably inherited.