Biomedical subjects

P Wehling

Publications and source records attributed to P Wehling.

At least 19 recordsLinked to original sources

PubMed2006

Mapping of Rym16Hb, the second soil-borne virus-resistance gene introgressed from Hordeum bulbosum.

Rym16(Hb), a gene conferring resistance to soil-borne viruses, was introgressed from Hordeum bulbosum to barley chromosome 2HL. Mechanical inoculation with BaMMV and field tests on a plot contaminated with different viruses demonstrated that Rym16(Hb) is effective against all European viruses of the soil-borne virus complex (BaMMV, BaYMV-1, -2). Genetic analysis revealed a dominant inheritance of the resistance controlled by Rym16(Hb). Using 2HL anchor markers, the size of the introgression was estimated to be about 30 M. In its proximal part, the introgression was characterized by a rearrangement of markers Xbcd266, ABC153 and ABC252, accompanied with pronounced linkage drag by factor 4 in segregating mapping populations. The introgression was found to be associated with a recessive lethality factor, l(Hb), which was closely linked to the markers mentioned above. Recombination occurring within the introgressed H. bulbosum segment allowed us to separate l(Hb) from Rym16(Hb) and to reduce the size of the introgression to 23 cM or less.

Chromosome Mapping↗

PubMed2005

Behaviour of Sinapis alba chromosomes in a Brassica napus background revealed by genomic in-situ hybridization.

Genomic in-situ hybridization (GISH) was applied to study the behaviour of addition chromosomes in first and second backcross (BC) progenies of hybrids between Brassica napus ssp. napus L. (AACC, 2n = 38) and Sinapis alba L. (SS, 2n = 24) produced by electrofusion. With GISH using genomic DNA of S. alba was used as probe it was possible to clearly distinguish both of the parental genomes and effectively monitor the fate of S. alba chromosomes in the BC(1) and BC(2) progenies. GISH analysis confirmed the sesquidiploid genome composition (AACCS) of the BC(1) progenies, which contained 38 chromosomes from B. napus and 12 chromosomes from S. alba. Genome painting in the pollen mother cells (PMCs) of the BC(1) plants revealed intergenomic association between B. napus and S. alba chromosomes, whereby a maximum of 4 trivalents between AC and S chromosomes were identified at metaphase I. In the BC(2) progenies, aneuploids with different numbers of additional chromosomes from S. alba, ranging from 1 to 7, were confirmed. Three putative monosomic alien addition lines were characterized, and the results are discussed with respect to the potential for intergenomic chromosome recombination.

Brassica napus↗

PubMed2005

Approaching the self-incompatibility locus Z in rye (Secale cereale L.) via comparative genetics.

Using barley and wheat expressed sequence tags as well as rice genomic sequence and mapping information, we revisited the genomic region encompassing the self-incompatibility (SI) locus Z on rye chromosome 2RL applying a comparative approach. We were able to arrange 12 novel sequence-tagged site (STS) markers around Z, spanning a genetic distance of 32.3 cM, with the closest flanking markers mapping at a distance of 0.5 cM and 1.0 cM from Z, respectively, and one marker cosegregating with Z, in a testcross population of 204 progeny. Two overlapping rice bacterial artifical chromosomes (BACs), OSJNBa0070O11 and OSJNBa0010D21, were found to carry rice orthologs of the three rye STS markers from the 1.5-cM interval encompassing Z. The STS-marker orthologs on these rice BACs span less than 125,000 bp of the rice genome. The STS marker TC116908 cosegregated with Z in a mapping population and revealed a high degree of polymorphism among a random sample of rye plants of various origin. TC116908 was shown via Southern hybridization to correspond to gene no. 10 (OSJNBa0070O11.10) on rice BAC OSJNBa0070O11. Reverse transcription-PCR with a TC116908-specific primer pair resulted in the amplification of a fragment of the expected size from the rye pistil but not from leaf cDNA. OSJNBa0070O11.10 was found to show a highly significant sequence similarity to AtUBP22, a ubiquitin-specific protease (UBP). TC116908 likely represents a putative UBP gene that is specifically expressed in rye pistils and cosegregates with Z. Given that the ubiquitination of proteins is emerging as a general mechanism involved in different SI systems of plants, TC116908 appears to be a promising target for further investigation with respect to its relation to the SI system of the grasses.

Blotting, Southern↗

PubMed2004

Leaf-rust resistance in rye (Secale cereale L.). 2. Genetic analysis and mapping of resistance genes Pr3, Pr4, and Pr5.

Three dominant resistance genes, Pr3, Pr4, and Pr5, were identified by genetic analysis of resistance to leaf rust in rye (Puccinia recondita f. sp. secalis). Each of the three genes confers resistance to a broad scale of single-pustule isolates (SPIs), but differences could be observed for specific Pr gene/SPI combinations. Resistance conferred by the three genes was effective in both detached-leaf tests carried out on seedlings and in field tests of adult plants. Molecular marker analysis mapped Pr3 to the centromeric region of rye chromosome arm 1RS, whereas Pr4 and Pr5 were assigned to the centromeric region of 1RL. Chromosomal localization and reaction patterns to specific SPIs provide evidence that the three Pr genes represent distinct and novel leaf-rust resistance genes in rye. The contributions of these genes to resistance breeding in rye and wheat are discussed.

Base Sequence↗

PubMed2004

Treatment of muscle injuries by local administration of autologous conditioned serum: animal experiments using a muscle contusion model.

Muscle contusions represent a major part of sports injuries. The suggested treatments are generally sufficient to support muscle healing, but require a relatively long period of time. Given that autologous blood products are safe treatments, we have used a technique which stimulates the release of certain growth factors in the autologous conditioned serum (ACS). Those growth factors are known to improve the proliferative activity of myogenic precursor cells. Mice were subjected to an experimental contusion injury to their gastrocnemius muscle; one group received local injections of ACS at 2 hrs, 24 hrs, and 48 hrs after injury, a control group received saline injections. The histology results showed that satellite cell activation at 30/48 hrs post injury was accelerated and the diameter of the regenerating myofibers was increased compared to the controls within the first week after injury. ELISA results on the ACS have shown that the elevations in FGF-2 (460 %) and TGF-beta1 (82 %) could be partly responsible for the accelerating effects on regeneration due to proliferative and chemotactic properties. We conclude that ACS injection is a promising approach to reduce the time of recovery from muscle injury. In terms of clinical targets, this new approach could be used in the treatment of sports injuries and may also be interesting in postoperative situations.

Animals↗

PubMed2004

Treatment of muscle injuries by local administration of autologous conditioned serum: a pilot study on sportsmen with muscle strains.

Muscle injuries represent a major part of sports injuries and are a challenging problem in traumatology. Strain injuries are the most common muscle injuries after contusions. These injuries can lead to significant pain and disability causing time to be lost to training and competition. Despite the frequency of strain injuries the treatment available is limited and is generally not sufficient to enhance muscle regeneration efficiently when fast resumption of sport activity is a primary target. A number of growth factors play a specific role in regeneration and it has been proven that a previously described method of physically and chemically stimulating whole blood (to produce autologous conditioned serum) induces concentration increases in FGF-2, HGF, and TGF-beta1. A preliminary study was conducted on muscle strain injuries in professional sportsmen receiving either: 1. autologous conditioned serum (ACS) or 2. Actovegin/Traumeel treatment as control. Assessment of recovery from injury was done by: 1. sport professional's ability to participate to 100 % under competition conditions in their respective sport and 2. MRI analysis. A significant difference in the recovery time from injury was demonstrated: 16.6 +/- 0.9 in the ACS treated instead of 22.3 +/- 1.2 (mean +/- SEM) days in the Actovegin/Traumeel control group (p = 0.001). MRI analysis supported the observed acceleration of the lesion recovery time. We conclude that ACS injection is a promising approach to reduce the time to recovery from muscle injury.

Athletic Injuries↗

PubMed2004

Cytogenetic characterization and fae1 gene variation in progenies from asymmetric somatic hybrids between Brassica napus and Crambe abyssinica.

Sexual progenies of asymmetric somatic hybrids between Brassica napus and Crambe abyssinica were analyzed with respect to chromosomal behavior, fae1 gene introgression, fertility, and fatty-acid composition of the seed. Among 24 progeny plants investigated, 11 plants had 38 chromosomes and were characterized by the occurrence of normal meiosis with 19 bivalents. The other 13 plants had more than 38 chromosomes, constituting a complete chromosomal set from B. napus plus different numbers of additional chromosomes from C. abyssinica. The chromosomes of B. napus and C. abyssinica origin could be clearly discriminated by genomic in situ hybridization (GISH) in mitotic and meiotic cells. Furthermore, meiotic GISH enabled identification of intergenomic chromatin bridges and of asynchrony between the B. napus and C. abyssinca meiotic cycles. Lagging, bridging and late disjunction of univalents derived from C. abyssinica were observed. Analysis of cleaved amplified polymorphic sequence (CAPS) markers derived from the fae1 gene showed novel patterns different from the B. napus recipient in some hybrid offspring. Most of the progeny plants had a high pollen fertility and seed set, and some contained significantly greater amounts of seed erucic acid than the B. napus parent. This study demonstrates that a part of the C. abyssinica genome can be transferred into B. napus via asymmetric hybridization and maintained in sexual progenies of the hybrids. Furthermore, it confirms that UV irradiation improves the fertility of the hybrid and of its sexual progeny via chromosomal elimination and facilitates the introgression of exotic genetic material into crop species.

Acetyltransferases↗

PubMed2003

Mapping of Rym14Hb, a gene introgressed from Hordeum bulbosum and conferring resistance to BaMMV and BaYMV in barley.

Hordeum bulbosum represents the secondary gene pool of barley and constitutes a potential source of various disease resistances in barley breeding. Interspecific crosses of H. vulgare x H. bulbosum resulted in recombinant diploid-barley progeny with immunity to BaMMV after mechanical inoculation. Tests on fields contaminated with different viruses demonstrated that resistance was effective against all European viruses of the soil-borne virus complex (BaMMV, BaYMV-1, -2). Genetic analysis revealed that resistance was dominantly inherited. Marker analysis in a F5 mapping family was performed to map the introgression in the barley genome and to estimate its size after several rounds of recombination. RFLP anchor-marker alleles indicative of an H. bulbosum introgression were found to cover an interval 2.9 cM in length on chromosome 6HS. The soil-borne virus resistance locus harboured by this introgressed segment was designated Rym14(Hb). For marker-assisted selection of Rym14(Hb) carriers, a diagnostic codominant STS marker was derived from an AFLP fragment amplified from leaf cDNA of homozygous-resistant genotypes inoculated with BaMMV.

Chromosome Mapping↗

PubMed2003

Leaf-rust resistance in rye (Secale cereale L.). 1. Genetic analysis and mapping of resistance genes Pr1 and Pr2.

Genetic analysis of resistance to leaf rust in rye (Puccinia recondita f. sp. secalis) led to the identification of two dominant resistance genes, Pr1 and Pr2. Both genes proved to be effective against a local leaf-rust population as well as a subset of single-pustule isolates (SPIs) the latter of which comprised SPIs with very high virulence complexity. Resistance conferred by Pr1 and Pr2 was expressed in detached-leaf tests of seedlings as well as in field tests of adult plants. Molecular marker analysis allowed us to map Pr1 in the proximal part of rye chromosome 6RL, whereas Pr2 was assigned to the distal part of chromosome 7RL. These results are discussed in view of homoeology relationships among Triticeae. A proposal is submitted for the designation of resistance genes to rye leaf rust which would avoid interference with existing gene-symboling in respect to wheat leaf-rust resistances introgressed from rye into wheat or triticale.

Basidiomycota↗

PubMed2003

The production of anti-inflammatory cytokines in whole blood by physico-chemical induction.

OBJECTIVE AND DESIGN: Cytokines such as interleukin-1 (IL-1) play an important role in degenerative musculo-skeletal diseases, including osteoarthritis, and a multitude of inflammatory disorders. Agents that inhibit the action of such cytokines have a high therapeutic potential in such diseases. Here we describe a new method for enhancing the production of the interleukin-l receptor antagonist (IL-1Ra) and other anti-inflammatory cytokines in whole blood. MATERIAL AND METHODS: Human venous blood was incubated in the presence of CrSO(4)-treated glass beads. Serum was recovered and the concentrations of IL-1Ra and other relevant cytokines were measured by ELISA. RESULTS: The interaction of the glass bead surface with cells in whole blood increased production of IL-1Ra and anti-inflammatory cytokines. Removal of the beads and centrifugation generated a serum preparation enriched in anti-inflammatory cytokines. This preparation is of therapeutic value in treating various inflammatory and degenerative disorders. CONCLUSIONS: The increased de novo production of anti-inflammatory cytokines by a direct physico-chemical induction of whole blood in the Orthokin system is feasible and offers an alternative, novel approach to treating mild to moderate OA and other orthopaedic conditions such as degenerative spine diseases.

Chemical Phenomena↗

PubMed2001

Transfer of genes to intervertebral disc cells: proposal for a treatment strategy of spinal disorders by local gene therapy.

Current treatments for spinal disease are unsatisfactory, and gene therapy holds promise as a means of ensuring prolonged and consistent delivery of therapeutic proteins into the spine. Direct injection of vectors into spinal structures is limited by the current lack of vectors with a satisfactory efficacy and safety profile. Conversely, ex vivo gene transfer into cells from the spine or other tissues (bone, nervous tissue, synovial membrane) followed by re-injection of these cells into the spine seems both appropriate and feasible in patients with degenerative disk disease. Candidate genes include genes encoding interleukin-1 antagonists, tumor necrosis factor antagonists, and growth factors. Further work is in order to move gene therapy research to the clinical trial stage in patients with degenerative disk disease, thus following in the footsteps of research on rheumatoid arthritis.

Animals↗

PubMed2000

Clinical trials in the gene therapy of arthritis.

Gene therapy clinical trials raise important safety issues that complicate their design and require extensive preclinical testing. Human protocols for the treatment of arthritis and most other orthopaedic and rheumatologic indications are complicated additionally by the perception that they are largely acquired, nonlethal conditions. Taking these considerations into account, the first such human study used the local, ex vivo delivery of a gene whose product, the interleukin-1 receptor antagonist, has an outstanding safety profile. This gene was delivered to the metacarpophalangeal joints of postmenopausal women 1 week before these joints were removed during total joint replacement surgery. In addition to providing an additional safety cushion, the surgical removal of the genetically modified joints made available large amounts of tissue to examine for evidence of successful gene transfer and gene expression. This Phase I safety study was approved at the local and federal levels, and its funding was contingent on the establishment of an external monitoring board. This trial now has been completed and a Phase II, efficacy study is being planned. A similar study has begun in Dusseldorf, Germany and results from the first two patients are similar to the results of the American patients. Permission has been given for two additional human trials, one in the United States and one in the Netherlands, in which a gene encoding herpes thymidine kinase will be transferred to the joints of patients with rheumatoid arthritis who then will be administered gancyclovir. This procedure aims to treat the disease by producing a genetic synovectomy. Additional development of human gene therapies for arthritis and other orthopaedic and rheumatic conditions will be aided by the successful completion of these studies.

Animals↗

PubMed1998

Spontaneous secretion of interleukin 1 receptor antagonist (IL-1ra) by cells isolated from herniated lumbar discal tissue after discectomy.

In the study presented, cells of a herniated lumbar disc were cultivated in vitro and analysed for interleukin 1beta (IL-1beta) and interleukin 1 receptor antagonist (IL-1Ra) production. The objective of this study was the detection of IL-1beta and IL-1Ra secreted by herniated lumbar discal cells after discectomy. The involvement of cytokines in the degeneration of intervertebral discs and in the pathophysiology of radiculopathy is established. Antagonizing proteins, e.g. IL-1Ra are thought to have considerable therapeutic potential. In the present study, a 51-year-old male with massive sequestrated lumbar disc herniation at L5/S1 was treated by microsurgical discectomy. Discal cells were isolated, cultures and culture supernatants immunochemically analysed for IL-1beta and IL-1Ra secretion. Spontaneous secretion of IL-1Ra was found. IL-1beta was not detected. Our findings might contradict recent studies on the role of IL-1beta and IL-1Ra. A possible therapeutic role of exogenous IL-1Ra in disc degeneration needs further research.

Cells, Cultured↗

PubMed1997

[Acupuncture together with cytokine depressing herbs in comparison to injection therapy with steroids in sciatic pain].

INTRODUCTION: Because of their frequency and consequences sciatica remains a demanding clinical entity with significant social and economical impact. There is a high demand on therapeutic modalities, using folk medicine methods in the treatment of orthopaedic diseases. No data is available on the usefulness of methods like natural herbs in combination with acupuncture. Aim of our study was to present data on the effect of acupuncture and cytokine-inhibiting natural herbs in sciatic pain. We compared these results with nerve root infiltration by local anaesthetics and corticosteroids in our orthopaedic outpatient clinic. MATERIALS AND RESULTS: 278 patients with chronic pain for at least 3 months were observed. All three therapeutical modalities showed improvement of pain scores. Best results were gained with steroid injection ( n=26; 66% pain reduction), acupuncture in combination with herbs improved the pain in average of 62% ( n=230), whereas injection with local anaesthetic had a pain reduction of 48% ( n=22). Sole Mepivacain-HCl treatment had significant less pain reduction compared to the other treatment modalities. CONCLUSION: According to our results acupuncture in combination with herbs and steroid nerve blocks appear to be an effective and thus recommendable conservative therapy in cases of sciatic pain. Taking into account that patients increasingly prefer ethnomedical modalities of treatment our study gains importance for practising orthopaedists.

English Abstract↗

PubMed1997

Transfer of genes to chondrocytic cells of the lumbar spine. Proposal for a treatment strategy of spinal disorders by local gene therapy.

STUDY DESIGN: In the current study, chondrocytic cells from bovine intervertebral end plates were cultivated in vitro and modified genetically. OBJECTIVE: The authors intended to perform isolation and cultivation of cells from bovine end plates of the spine. They also intended to show, in principle, the feasibility of introducing exogenous genes into chondrocytic cells from bovine intervertebral end plates by way of retroviral vectors. SUMMARY OF BACKGROUND DATA: The involvement of cytokines in the destruction of articular cartilage is established. It appears possible that similar mechanisms may play a role in intervertebral disc degeneration and other spinal disorders. Conventional medication and surgery of intervertebral disc degeneration addresses neither the pathophysiology nor the chronicity of the disease. Therapeutic proteins carry great potential as locally produced drugs after transfer of their cognate genes to the sites of interest. METHODS: Vertebral end plate tissue was obtained from bovine os coccygis. Chondrocytic cells were isolated and cultured in vitro. The bacterial beta-galactosidase (LacZ) gene and, alternatively, the complementary DNA (DNA copy of the mRNA) of the human interleukin-1 receptor antagonist were introduced into the isolated cells by retrovirus mediated gene transfer. beta-galactosidase activity was determined by staining, and interleukin-1 receptor antagonist protein was quantitated by enzyme-linked immunosorbent assay. RESULTS: Isolation and cultivation of chondrocytic end plate cells is possible. Native cells continue to grow in culture for more than 2 months. Transfer of the beta-galactosidase gene to cultured cells resulted in approximately 1% beta-galactosidase positive cells. Transfer of the interleukin-1 receptor antagonist complementary DNA resulted in the production of 24 ng/ml/10(6) cells interleukin-1 receptor antagonist protein in 48 hours. CONCLUSIONS: The introduction of exogenous therapeutic genes into cells from the intervertebral end plate opens the possibility for a local gene-based treatment of intervertebral disc degeneration. This approach avoids some of the shortcomings of conventional drug- and surgery-based treatments and has the potential to be specific, effective, and appropriate to the chronicity of the disease.

Animals↗

PubMed1997

Growth changes of solidly fused kyphotic bloc after surgery for tuberculosis. Comparison of four procedures.

STUDY DESIGN: A study to analyze the changes of the spinal deformity during the growth period, with regard to different operations for spinal tuberculosis in children. OBJECTIVES: To quantify the changes in the kyphotic angle and the growth ratio of the fusion bloc during spinal growth for different fusion techniques. SUMMARY OF BACKGROUND DATA: Most of the publications dealing with spinal tuberculosis in children focused on the clinical outcome with regard to different conservative and operative treatments. There is little reliable information concerning the growth of the solidly fused kyphotic bone bloc and its influence on the changes of the kyphotic deformity after different operative procedures. METHODS: The study included 117 children operated on for spinal tuberculosis at the age of 2-6 years at the Ruttonjee Sanatorium in Hong Kong during the 1950s and 1960s. Lateral radiographs obtained postoperatively and 5 and 10 years after the operation were analyzed for the growth changes of the solidly fused bone bloc. These results were compared with the different operation techniques (e.g., anterior fusion, posterior fusion, combined anterior and posterior fusion, and anterior debridement without fusion). RESULTS: The patients treated by anterior fusion showed the worst results with respect to the kyphotic angle. This was especially true when the lesion was located in the thoracic spine and several segments were involved. Regarding the growth ratio of the fusion bloc, only the combined fusion and the anterior debridement guaranteed an equal growth of the anterior and posterior height. CONCLUSIONS: Radical anterior surgery for spinal tuberculosis destroys the anterior growth and limits the capacity for spinal remodeling. Therefore, it should be avoided, if it is not absolutely necessary, for the healing of the infection or the primary correction of the tuberculous deformity.

Bone Remodeling↗

PubMed1996

Neurophysiologic changes in lumbar nerve root inflammation in the rat after treatment with cytokine inhibitors. Evidence for a role of interleukin-1.

STUDY DESIGN: The present study was designed to evaluate the effect of cytokine inhibitors in experimental allergic radiculitis. OBJECTIVE: Evaluation of the effect of cytokine inhibitors in experimental allergic radiculitis. SUMMARY OF BACKGROUND DATA: A number of cytokines are known to be involved in hyperalgesia and may play a role in radiculitis. Corticosteroids and other cytokine inhibitors antagonize their effects. METHODS: Experimental allergic radiculitis was induced in rats by injection of bovine myelin from the peripheral nervous system. The sham group subsequently received saline injections; the treatment groups received either prednisolone or interleukin-1 receptor antagonist. Treatment effect was assessed on the basis of motor performance and neurophysiologic parameters. RESULTS: Treatment ameliorated the symptoms of experimental allergic radiculitis. Prednisolone appeared to be somewhat more effective than interleukin-1 receptor antagonist. CONCLUSIONS: Because interleukin-1 receptor antagonist specifically blocks the effects of interleukin-1 at its receptors, the present results imply that interleukin-1 is a causal factor in the model of experimental radiculitis used. Its specificity and apparent lack of side effects make interleukin-1 receptor antagonist an attractive candidate treatment for the human disease.

Animals↗

PubMed1995

Magnetic stimulation as a diagnostic tool in cervical nerve root compression and compression-induced neuropathy.

Magnetic stimulation of motor cortex and cervical spinal cord was used to assess motor impairment in patients with either cervical nerve root compression or myelopathy. Evoked potentials were recorded from biceps brachii and abductor pollicis brevis. Our interest centered on whether the paresis resulting from root compression is accompanied by an increased latency of magnetically evoked muscle potentials and whether latency is increased in cases of cervical root compression or myelopathy in which no paresis occurs. Latency increase does appear to be a moderately good indicator of disturbed nerve conduction. In particular, prolonged latencies in cases of "subclinical" paresis could be used as an important diagnostic tool for the early detection of motor deficits in cervical compression radiculopathy and myelopathy.

Electrodiagnosis↗