Biomedical subjects

A Lahm

Publications and source records attributed to A Lahm.

At least 19 recordsLinked to original sources

PubMed2006

Regression and correlation analysis of preoperative versus intraoperative assessment of axes during navigated total knee arthroplasty.

OBJECTIVE: Modern computer assisted surgery (CAS) systems allow accurate positioning of the implants in navigated Total Knee Arthroplasty (TKA). However, when an operation is performed with a navigation system, it is important to know if the anatomical situation of the knee is reflected equally in both the preoperative image (e.g., CT) and the intraoperative navigation setup. In this study, we compared the preoperative anatomical situation to the virtual intraoperative situation of the navigation setup. MATERIAL AND METHODS: We analyzed 24 navigated operations. Intraoperatively, the condylar twist angle (CTA) was documented with the navigation system by measuring the angle between the transepicondylar axis (TEA) and posterior condyle axis (PCA). This data was compared with the preoperative data from the CT scan. RESULTS: Statistical analysis revealed that there was no correlation between the pre- and intraoperative data (r = 0.095). CONCLUSIONS: Statistically, there is no possibility of collecting the same angles and axes when using the two different methods (CT and navigation) on the same knee. It is not possible to copy the preoperative anatomical situation exactly with the virtual intraoperative data. Reasons for this include systematic errors, as well as inter- and intraobserver errors in both methods.

Arthroplasty, Replacement, Knee↗

PubMed2005

Subchondral and trabecular bone remodeling in canine experimental osteoarthritis.

INTRODUCTION: We wanted to test the hypothesis that quality changes occur in early-stage arthritic subchondral cancellous bone after acute subchondral damage. So far, not much attention has been paid to changes of the subchondral bone after traumatic subchondral lesions. MATERIALS AND METHODS: With an established animal model, we produced pure subchondral damage without initial affection of the articular cartilage in 12 Beagle dogs under MRI and histological control. We utilized bone histomorphometry to evaluate bone turnover, its structure and the articular cartilage 6 months after the initial damage. RESULTS: On follow-up, bone remodelling was indicated, e.g. by a significant increase in the trabecular bone volume and thickness, osteoblast number and osteoid surface and a decrease in the trabecular number in all 12 samples. Several other parameters showed a tendency, e.g. osteoblast surface and osteoclast number. Cartilage analysis showed degenerative changes in ten of 12 samples that had not shown any evidence of damage during the initial examination. DISCUSSION: Our investigation indicates a significant deterioration in the architecture of the cancellous bone with degenerative changes of the overlying articular cartilage after subchondral lesions, which change the mechanical properties.

Animals↗

PubMed2005

Subchondral bone trauma causes cartilage matrix degeneration: an immunohistochemical analysis in a canine model.

UNLABELLED: Joint instability was believed to be the main cause of osteoarthritis following non-fracture articular trauma. However, sudden high impact load through articular cartilage onto subchondral bone may also cause osteoarthritic changes. OBJECTIVE: We asked whether early osteoarthritic changes following transarticular impact may be depicted using immunofluorescence on unfixed cryosections to contribute to a more detailed understanding of degenerative processes of joint impaction. DESIGN: Transarticular impacts were applied to patellofemoral joints of 12 skeletally mature beagle dogs (age: 15-16 months) using a drop tower. Biopsies of impact areas were sampled after 6 months and processed for standard light microscopy on formalin-fixed sections and for immunofluorescence for collagen type I (col I), type II (col II) and aggrecan (AC) on unfixed cryosections. Gross morphology and immunofluorescence on cryosections were documented using a semi-quantitative scaling system, compared to healthy controls and to standard light microscopy. RESULTS: Four biopsies showed almost entirely fibrocartilaginous morphology, four appeared to be of preserved hyaline morphology with only minor signs of fibrocartilaginous remodelling and four showed preserved hyaline appearance. We found decrease in col II and AC expression in highly degenerative specimens as well as increase of col I expression. Increased col I expression in the pericellular matrix could even be depicted in specimens with intact hyaline morphology. DISCUSSION/CONCLUSION: Observations suggest that joint impaction causes early osteoarthritic changes after 6 months. Collagen network disruption seems to lead to AC loss, although other researchers found isolated AC loss without denaturation of col II using immunofluorescence in formalin-fixed specimens. This is the first study on effects of transarticular impact using immunofluorescence on unfixed cryosections.

Aggrecans↗

PubMed2005

An experimental canine model for subchondral lesions of the knee joint.

Aim of the study was to create an animal model for the investigation of the role of subchondral bone damage without initial cartilage lesion in the pathogenesis of osteoarthritis, the mechanical properties of the joints as well as its role in cartilage metabolism. Therefore, after cadaver studies an animal model was created to apply a transarticular load to the femoro-patellar joint under reproducible conditions and produce a pure subchondral damage without affecting the articular cartilage. Following the cadaver studies a first group of four dogs was impacted to identify forces to produce isolated subchondral fractures in the femoral condyle. Then a second group of 12 dogs knee joints was impacted under identical conditions with forces of approximately 2100 N to produce similar subchondral fractures without cartilage damage in one joint under MRI control: T1-weighted SE-sequences. T2-weighted TSE, fat suppressed TIRM-sequences and 3D-FLASH fat saturated sequences. FLASH 3D-sequences revealed intact cartilage after impact in all cases and TIRM-sequences showed subchondral fractures representing bleeding, microfractures and fragmented bone trabecules. Turbo spin echo sequences and T1-weighted images revealed other intact intraarticular structures such as ligaments and menisci. The proposed experimental animal model is suitable to investigate the effect of pure subchondral damage on the articular cartilage and on means of treatment of cartilage defects without surgical intervention and without initial cartilage damage.

Animals↗

PubMed2005

Experimental autologous osteochondral plug transfer in the treatment of focal chondral defects: magnetic resonance imaging signs of technical success in sheep.

PURPOSE: To describe the magnetic resonance imaging (MRI) signs of technically successful osteochondral plug transfer and to correlate the findings with histology using the Mankin score. MATERIAL AND METHODS: The study was done in a prospective animal experiment: 11 adult black-head sheep underwent surgical treatment with osteochondral plug transfer of a knee joint. The animals were killed 6 months later and MRI of the joints was done immediately. MRI was applied with a 1.5T MR scanner using a spin-echo (SE) T1-weighted, turbo spin-echo (TSE) T2-weighted with spectral fat suppression and a fat-suppressed 3D-spoiled gradient echo (GRE) sequence (manufacturer's acronym: FLASH) (TR 50.0 ms, TE 11.0 ms, flip 35 degrees). After MRI, all knee joints were dissected and a biopsy of the plug and the adjacent cartilage was taken. Classification of the cartilage biopsies was carried out in accordance with a modified Mankin score. RESULTS: Cartilage repairs with a hypointense cartilage signal in the FLASH 3D sequence were correlated with poor histological results (lower Mankin score). Histologically, the regions of cartilage with a hypointense signal showed a fibrocartilage-like repair tissue. Hyaline cartilage with well-defined layers had the same signal intensity in the FLASH sequence relative to adjacent hyaline cartilage. There were two plugs with a surface defect, graded as Outerbridge grade 1 in MRI and histology. Both had a poor outcome in the histologic Mankin score. Grade 2-4 lesions were not observed in the MRI study nor in the histologic study. CONCLUSION: MRI is a useful non-invasive tool for evaluating the morphologic status of osteochondral plug transfers. A good postoperative result of the cartilage repair was found histologically if an isointense cartilage signal of the graft was documented in the FLASH 3D sequence, and the graft had good congruity with the articular surface without defects.

Animals↗

PubMed2005

Comparison of total and partial traumatic tears of the subscapularis tendon.

We treated surgically 16 shoulders with an isolated traumatic rupture of the subscapularis tendon over a six-year period. Nine patients had a total and seven a partial tear of the subscapularis tendon. Repair was undertaken through a small deltopectoral groove approach. The mean Constant score improved in total tears from 38.7 to 89.3 points (p = 0.003) and in partial tears from 50.7 to 87.9 points (p = 0.008). The total tears were significantly more improved by surgery than the partial tears (p = 0.001). The delay between trauma and surgery was inversely proportional to the improvement in the Constant score suggesting that early diagnosis and surgical repair improves outcome.

Adult↗

PubMed2004

Photoshop-based image analysis of canine articular cartilage after subchondral damage.

INTRODUCTION: The validity of histopathological grading is a major problem in the assessment of articular cartilage. Calculating the cumulative strength of signal intensity of different stains gives information regarding the amount of proteoglycan, glycoproteins, etc. Using this system, we examined the medium-term effect of subchondral lesions on initially healthy articular cartilage. MATERIALS AND METHODS: After cadaver studies, an animal model was created to produce pure subchondral damage without affecting the articular cartilage in 12 beagle dogs under MRI control. Quantification of the different stains was provided using a Photoshop-based image analysis (pixel analysis) with the histogram command 6 months after subchondral trauma. RESULTS: FLASH 3D sequences revealed intact cartilage after impact in all cases. The best detection of subchondral fractures was achieved with fat-suppressed TIRM sequences. Semiquantitative image analysis showed changes in proteoglycan and glycoprotein quantities in 9 of 12 samples that had not shown any evidence of damage during the initial examination. Correlation analysis showed a loss of the physiological distribution of proteoglycans and glycoproteins in the different zones of articular cartilage. CONCLUSION: Currently available software programs can be applied for comparative analysis of histologic stains of hyaline cartilage. After subchondral fractures, significant changes in the cartilage itself occur after 6 months.

Animals↗

PubMed2002

A model for recognition of polychlorinated dibenzo-p-dioxins by the aryl hydrocarbon receptor.

Ligand binding by the aryl hydrocarbon receptor (AhR), a member of the bHLH-PAS family of transcriptional regulatory proteins, has been mapped to a region within the second 'PAS' domain, a conserved sequence motif first discovered in the Per-ARNT-Sim family of proteins. In addition to the bacterial photoactive yellow protein (PYP), which had been proposed as a structural prototype for the three dimensional fold of PAS domains, two crystal structures of the PAS domain have recently been determined: the human potassium channel HERG and the heme binding domain of the bacterial O(2) sensing FixL protein. The three structures reveal a highly conserved structural framework in evolutionary rather distant PAS domains, provide a more general view of how these domains can recognize their ligands and suggest a structure-function relationship that we exploited to build a three-dimensional model of the ligand binding domain (LBD) of the mouse aryl hydrocarbon receptor (mAhR). The model allowed us to putatively identify the residues responsible for the recognition of polychlorinated dibenzo-p-dioxins (PCDDs) by AhR receptors and to formulate an hypothesis on the signal transduction mechanism.

Amino Acid Sequence↗

PubMed2002

Hepatitis C virus proteins as targets for drug development: the role of bioinformatics and modelling.

Hepatitis C virus (HCV), a member of the Flaviviridae family, has been recognised to be responsible for both parenterally transmitted and sporadic non-A and non-B hepatitis affecting 1-3% of the world population. HCV is a positive stranded RNA virus encoding a single polyprotein which contains at least ten unique structural and non-structural proteins. Amongst these the structural protein E2 has been of special interest for vaccine development and the serine protease NS3, which is responsible for cleavage of the polyprotein, for the development of small molecule inhibitors. We will focus on the contribution of computational techniques and the use of structural information for the design and discovery of novel therapeutic agents for these targets. Both drug discovery and vaccine design efforts will be discussed taking into account also the problem of emerging resistance.

Computational Biology↗

PubMed2001

Mimotopes of the hyper variable region 1 of the hepatitis C virus induce cross-reactive antibodies directed against discontinuous epitopes.

Hepatitis C virus (HCV) is a major cause worldwide of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma, and the development of an effective vaccine represents a high priority goal. The hyper variable region 1 (HVR1) of the second envelope protein (E2) of HCV contains a principal neutralizing determinant, but it is highly variable among different isolates and it is involved in the escape from host immune response. To be effective, a vaccine should elicit a cross-reacting humoral response against the majority of viral variants. We show that it is possible to achieve a broadly cross-reactive immune response in rabbits by immunization with mimotopes of the HVR1, selected from a specialized phage library using HCV patients' sera. Some of the cross-reacting anti-mimotope antibodies elicited in rabbits, recognize discontinuous epitopes in a manner similar to those induced by the virus in infected patients.

Amino Acid Sequence↗

PubMed2001

Characterization of the hepatitis C virus NS2/3 processing reaction by using a purified precursor protein.

The NS2-NS3 region of the hepatitis C virus polyprotein encodes a proteolytic activity that is required for processing of the NS2/3 junction. Membrane association of NS2 and the autocatalytic nature of the NS2/3 processing event have so far constituted hurdles to the detailed investigation of this reaction. We now report the first biochemical characterization of the self-processing activity of a purified NS2/3 precursor. Using multiple sequence alignments, we were able to define a minimal domain, devoid of membrane-anchoring sequences, which was still capable of performing the processing reaction. This truncated protein was efficiently expressed and processed in Escherichia coli. The processing reaction could be significantly suppressed by growth in minimal medium in the absence of added zinc ions, leading to the accumulation of an unprocessed precursor protein in inclusion bodies. This protein was purified to homogeneity, refolded, and shown to undergo processing at the authentic NS2/NS3 cleavage site with rates comparable to those observed using an in vitro-translated full-length NS2/3 precursor. Size-exclusion chromatography and a dependence of the processing rate on the concentration of truncated NS2/3 suggested a functional multimerization of the precursor protein. However, we were unable to observe trans cleavage activity between cleavage-site mutants and active-site mutants. Furthermore, the cleavage reaction of the wild-type protein was not inhibited by addition of a mutant that was unable to undergo self-processing. Site-directed mutagenesis data and the independence of the processing rate from the nature of the added metal ion argue in favor of NS2/3 being a cysteine protease having Cys993 and His952 as a catalytic dyad. We conclude that a purified protein can efficiently reproduce processing at the NS2/3 site in the absence of additional cofactors.

Amino Acid Sequence↗

PubMed2001

Induction of cross-reactive humoral immune response by immunization with mimotopes of the hypervariable region 1 of the hepatitis C virus.

Hepatitis C Virus (HCV) is a major cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma, worldwide, and the development of an effective vaccine represents a high priority goal. The Hyper Variable Region 1 (HVR1) of the second Envelope protein (E2) of HCV contains a principal neutralizing determinant, but it is highly variable among different isolates and it is involved in the escape from host immune response. Thus, to be effective, a vaccine should elicit a cross-reacting humoral response against the majority of viral variants. We show that it is possible to achieve a broadly cross-reactive immune response in rabbits by immunization with mimotopes of the HVR1. selected from a specialized phage library using HCV patients' sera. At least some of the cross-reacting anti-mimotope antibodies, elicited in rabbits, recognize discontinuous epitopes in a manner similar to those induced by the virus in infected patients.

Amino Acid Sequence↗

PubMed2000

A model for the hepatitis C virus envelope glycoprotein E2.

Several experimental studies on hepatitis C virus (HCV) have suggested the envelope glycoprotein E2 as a key antigen for an effective vaccine against the virus. Knowledge of its structure, therefore, would present a significant step forward in the fight against this disease. This paper reports the application of fold recognition methods in order to produce a model of the HCV E2 protein. Such investigation highlighted the envelope protein E of Tick Borne Encephalitis virus as a possible template for building a model of HCV E2. Mapping of experimental data onto the model allowed the prediction of a composite interaction site between E2 and its proposed cellular receptor CD81, as well as a heparin binding domain. In addition, experimental evidence is provided to show that CD81 recognition by E2 is isolate or strain specific and possibly mediated by the second hypervariable region (HVR2) of E2. Finally, the studies have also allowed a rough model for the quaternary structure of the envelope glycoproteins E1 and E2 complex to be proposed. Proteins 2000;40:355-366.

Amino Acid Sequence↗

PubMed2000

A new determinant of endoplasmic reticulum localization is contained in the juxtamembrane region of the ectodomain of hepatitis C virus glycoprotein E1.

Hepatitis C virus glycoproteins E1 and E2 do not reach the plasma membrane of the cell but accumulate intracellularly, mostly in the endoplasmic reticulum. Previous studies based on transient expression assays have shown that the transmembrane domains of both glycoproteins are sufficient to localize reporter proteins in the endoplasmic reticulum and that other localization signals may be contained in the ectodomain of E1 protein. To identify such signals we generated chimeric proteins between E1 and two reporter proteins, the human CD8 glycoprotein and the human alkaline phosphatase, and analyzed their subcellular localization in stable as well as transient transfectants. Our results showed that (i) an independent localization determinant for the endoplasmic reticulum is present in the juxtamembrane region of the ectodomain of E1 protein and (ii) the localization dictated by this determinant is either due to direct retention or to a recycling mechanism from the intermediate compartment/cis-Golgi complex region, which is clearly different from those previously described for other retrieval signals. These results show for the first time in mammalian cells that the localization in the endoplasmic reticulum of transmembrane protein can be determined by specific targeting signals acting in the lumen of the compartment.

Amino Acid Sequence↗

PubMed2000

Bacteriophage lambda display of complex cDNA libraries: a new approach to functional genomics.

We describe the construction and characterization of two lambda surface displayed cDNA expression libraries derived from human brain and mouse embryo. cDNA inserts were obtained by tagged random-priming elongation of commercially available cDNA libraries and cloned into a novel lambda vector at the 3' end of the D capsid protein gene, which produced highly complex repertoires (1x10(8) and 2x10(7) phage). These libraries were affinity selected with a monoclonal antibody against the neural specific factor GAP-43 and with polyclonal antibodies that recognize the EMX1 and EMX2 homeoproteins. In both cases rapid identification of specific clones was achieved, which demonstrates the great potential of the lambda display system for generating affinity selectable cDNA libraries from complex genomes.

Amino Acid Sequence↗

PubMed2000

Arthroscopic management of osteochondral lesions of the talus: results of drilling and usefulness of magnetic resonance imaging before and after treatment.

Since the advent of operative ankle arthroscopy and magnetic resonance imaging (MRI) specific treatment of osteochondritis dissecans of the talus has progressed rapidly. Drilling is still the treatment of choice in early stages of osteochondritis dissecans of the talus. Rear-entry guides and preoperative planning with MRI have led to better results with this kind of treatment. Within 5 years, 42 patients (26 male and 16 female) underwent arthroscopic treatment of osteochondritis dissecans of the talus, 22 underwent percutaneous drilling, 13 cancellous bone grafting, 4 refixation, and 3 curettage. The average age of the patients was 28 years (range, 11 to 53 years). A clinical score system was used in a clinical and MRI follow-up of 19 of the patients with K-wire drilling. Up to 100 points are given in the categories pain, stability/insecurity, efficiency/pain-free walking distance, gait, differences in circumference, range of motion, and power. There was a history of trauma in 31 of the 42 patients. The majority of lesions (24 cases) were localized at the lateral talus, and these patients all had trauma. In 11 of the 18 lesions at the medial talus, there was no evidence of trauma. The 19 patients in the follow-up achieved an average of 87 points. K-wire drilling represents the chief component of early stages with intact or partially fractured cartilage surface, whereas arthroscopically controlled cancellous bone grafts after curettage are used in grade II stages only. Results of K-wire drilling are not worse than those of cancellous bone grafts; this is attributable to a generous perforation of the sclerosis. This has contributed to an improved preoperative diagnosis with MRI.

Adolescent↗

PubMed2000

Mutational analysis of hepatitis C virus NS3-associated helicase.

Nonstructural protein 3 (NS3) of hepatitis C virus contains a bipartite structure consisting of an N-terminal serine protease and a C-terminal DEXH box helicase. To investigate the roles of individual amino acid residues in the overall mechanism of unwinding, a mutational-functional analysis was performed based on a molecular model of the NS3 helicase domain bound to ssDNA, which has largely been confirmed by a recently published crystal structure of the NS3 helicase-ssDNA complex. Three full-length mutated NS3 proteins containing Tyr(392)Ala, Val(432)Gly and Trp(501)Ala single substitutions, respectively, together with a Tyr(392)Ala/Trp(501)Ala double-substituted protein were expressed in Escherichia coli and purified to homogeneity. All individually mutated forms showed a reduction in duplex unwinding activity, single-stranded polynucleotide binding capacity and polynucleotide-stimulated ATPase activity compared to wild-type, though to different extents. Simultaneous replacement of both Tyr(392) and Trp(501) with Ala completely abolished all these enzymatic functions. On the other hand, the introduced amino acid substitutions had no influence on NS3 intrinsic ATPase activity and proteolytic efficiency. The results obtained with Trp(501)Ala and Val(432)Gly single-substituted enzymes are in agreement with a recently proposed model for NS3 unwinding activity. The mutant phenotype of the Tyr(392)Ala and Tyr(392)Ala/Trp(501)Ala enzymes, however, represents a completely novel finding.

Adenosine Triphosphatases↗

PubMed1999

[Clinical and cell biology aspects of autologous chondrocytes transplantation].

The treatment of deep cartilage defects is a challenge for every orthopaedic surgeon. The potential for regeneration of cartilage tissue is minimal and leads to mechanically inferior fibrous tissue. The established techniques induce the growth of fibrous tissue but fail to prevent arthrosis. Autologous chondrocyte transplantation seems to be the most promising therapy concept with clinical relevance to reserves a full thickness cartilage defect with hyaline-like cartilage. Outcome studies with a follow up from 2-10 years show in up to 90 % good and excellent results for defects on the femoral condyle and 70 % for the patella. Mechanical testing of the regenerated cartilage showed almost similar stiffness as nearly normal hyaline cartilage. The available data justify the acceptance of autologous chondrocyte transplantation as a standard procedure for limited indications and well-trained surgeons. Result of already inaugurated studies will show the potential of chondrocyte transplantation to prevent osteoarthritis.

Animals↗