Integrative promoter cloning plasmid vectors for Rhizobium meliloti.

A set of integrative 'promoter probe' plasmids were constructed for both translational and transcriptional fusions. The vectors are based on the broad host range, low copy number plasmid pRK290 (IncPl) in which the attachment site of Rhizobium phage 16-3 and the lacZ gene of Escherichia coli were combined. The vectors integrate into the chromosome of Rhizobium meliloti, providing also the advantages of the single copy promoter probe cassettes. Thus they fulfil the prerequisite of the systems used for investigating gene regulation. The plasmids were applied for the study of the transcription regulation of the 16-3 phage. Their versatile use is also demonstrated.

Long-term effects of toe transfers on the donor feet.

The purpose of this study was to assess the long-term consequences to the donor site of microvascular transfer of one or two toes. This was achieved retrospectively by patient-reported, clinician-based and functional testing. Weight-bearing radiographs were taken of both feet. Seventy-four patients (80 feet, 84 transferred toes) participated in this study. After a median of 16 years follow-up, most patients reported no or only minor complaints concerning the donor site. Cold intolerance and pain during exertion were the most commonly reported complaints. According to the two functional scores used, 92% of patients reported no or minor complaint and 83% of patients received a good result at the donor feet. Maintaining first ray alignment and avoiding early postoperative complications predicted a better outcome. Even though donor site ramifications should be expected after microvascular transfer of toes, patient satisfaction remains high.

Adverse reaction to metal debris is more common in patients following MoM total hip replacement with a 36 mm femoral head than previously thought: results from a modern MoM follow-up programme.

We conducted a retrospective study to assess the prevalence of adverse reactions to metal debris (ARMD) in patients operated on at our institution with metal-on-metal (MoM) total hip replacements with 36 mm heads using a Pinnacle acetabular shell. A total of 326 patients (150 males, 175 hips; 176 females, 203 hips) with a mean age of 62.7 years (28 to 85) and mean follow-up of 7.5 years (0.1 to 10.8) participating in our in-depth modern MoM follow-up programme were included in the study, which involved recording whole blood cobalt and chromium ion measurements, Oxford hip scores (OHS) and plain radiographs of the hip and targeted cross-sectional imaging. Elevated blood metal ion levels (> 5 parts per billion) were seen in 32 (16.1%) of the 199 patients who underwent unilateral replacement. At 23 months after the start of our modern MoM follow-up programme, 29 new cases of ARMD had been revealed. Hence, the nine-year survival of this cohort declined from 96% (95% CI 95 to 98) with the old surveillance routine to 86% (95% CI 82 to 90) following the new protocol. Although ARMD may not be as common in 36 mm MoM THRs as in those with larger heads, these results support the Medicines and Healthcare Products Regulatory Agency guidelines on regular reviews and further investigations, and emphasise the need for specific a follow-up programme for patients with MoM THRs.

Long-term functional results of microvascular toe-to-thumb reconstruction.

The purpose of this study was to evaluate the long-term functional results of microvascular toe-to-thumb reconstruction after trauma. Forty-one patients meeting the inclusion criteria were available for a clinical follow-up study. The function of the hand was assessed with questionnaires as well as with modified Tamai and Sollerman hand function tests. According to the questionnaires, most activities were considered easy or quite easy and the majority of the patients (36/41) managed with no or minor complaints. Clinical tests showed good recovery of function. Patient satisfaction was high. There were superficial infections in five hands and in six donor feet. In total, 16 late corrective operations were done to eight patients. Microvascular toe transfer is a good option for grip reconstruction after thumb amputation. The extent of the initial injury influences the achievable outcome, yet even a single toe transfer can restore adequate grip function.

The functional results of post-traumatic metacarpal hand reconstruction with microvascular toe transfers.

The aim of this retrospective study was to evaluate the functional results of grip reconstruction after metacarpal amputation with microvascular toe transfer or transfers. The Sollerman hand function test and modified Tamai score were determined. Additionally, secondary objective outcomes were measured. As subjective outcomes, the patients were asked about pain, satisfaction, sensibility and activities of daily living (ADL). Complications and secondary operations were also recorded. An average of 12 years 5 months (range 11-270 months) follow-up of 8 patients (11 transfers) is presented. Sollerman hand function test averaged 54, range 36-73. The modified Tamai score averaged 63, range 54-70. All except one patient were either satisfied or highly satisfied. Patients' perception of function in terms of ADL was generally good (average 26 out of 44) with many activities causing no difficulty or only slight difficulty. The study showed that it is possible to reconstruct a reasonable grip using microvascular toe transfers.

Footprinting studies of specific complexes formed by RepA, a replication initiator of plasmid pCU1, and its binding site.

The basic replicon of plasmid pCU1 contains three different replication origins. Replication initiated from the oriB origin requires pCU1-encoded protein RepA. Previously, information analysis of 19 natural RepA binding sequences predicted a 20-bp sequence as a RepA binding site. Guanines contacting RepA in the major groove of DNA have also been determined. In this study, we used the missing-nucleoside method to determine all of the bases relevant to RepA binding. The importance of some thymine bases was also confirmed by a missing-thymine site interference assay. Participation of the 5-methyl groups of two thymines (at positions -6 and 7) in RepA binding was pointed out by a missing-thymine methyl site interference assay. Phosphate groups of the DNA backbone which strongly interfered with RepA binding upon ethylation were also identified. The pattern of contacting positions mapped by hydroxyl radical protection footprinting indicates that RepA binds to one face of B-form DNA. The length of the binding site was found to be 20 bp by dissociation rate measurement of complexes formed between RepA and a variety of binding sequences. The symmetry of the binding site and that of the contacting bases, particularly the reacting 5-methyl groups of two thymines, suggest that pCU1-encoded RepA may contact its site as a homodimer.

Four new inverted terminal repeat sequences from bovine adenoviruses reveal striking differences in the length and content of the ITRs.

The inverted terminal repeat (ITR) of the genome of four bovine adenovirus (BAdV) types have been sequenced, analysed and compared to the ITRs of other adenoviruses. The length of ITRs of the examined BAdVs ranged between 59 and 368 base pairs, thus the presently known longest adenovirus ITR sequence is from BAdV-10. The conserved motifs and characteristic sequence elements of the ITRs providing different binding sites for replicative proteins of viral and cellular origin seemed to be distributed according to the proposed genus classification of BAdVs. The ITRs of BAdV-10 share similarity with the members of the genus Mastadenovirus, while the ITRs of the other three sequenced serotypes (BAdV-4, 5 and strain Rus) which are candidate members of the genus Atadenovirus are very short and contain NFI and Sp1 binding sites only. The analysis of the new ITRs implied that the nucleotide sequence of the so-called core origin is highly preserved within the mastadenovirus genus only.