Total knee replacement in triple deformity with posterior subluxation of the knee joint.

Dislocation or subluxation following total knee arthroplasty has been extensively reported, but vice versa that is total knee replacement for subluxed or dislocated knee has not been published. Triple deformity of knee that is flexion, external rotation, valgus at knee associated with posterior subluxation of tibia occurs in rheumatoid arthritis, advanced tubercular arthritis and neglected posttraumatic residual dislocated knee. A 50 year old female with seropositive rheumatoid arthritis had the above disabling deformity in left lower limb and varus with medial tibial thrust in the other. Bilateral total knee arthroplasty was planned. Conservative method of reduction of left knee posterior subluxation preoperatively by 90-90 skeletal traction failed; hence patient was subjected to a staged bilateral total knee replacement using an innovative technique. The most difficult and determining initial surgical step of knee replacement in such dislocated/subluxed knee is reduction of posterior subluxation and gaining flexion at knee, as only after gaining flexion and reducing dislocated tibia, will we be able to do knee arthroplasty in triple deformity of knee. These knees are grossly unstable as most of the capsule-ligamentous structures are attritioned/non-existent. So, a fine balance of bone cuts and soft-tissue release needs to be done in a sequential manner to fine tune valgus and posterior subluxation correction without jeoparadising neurovascular structures. After 3 years of knee arthroplasty the patient has painless, stable knee with good range of motion and is able to do all her activities, of living in a hilly terrain.

Streptococcus pneumoniae serotype 6C: an intra- and interclonal complex comparison.

We report the annotated draft genome sequences of four serotype 6C Streptococcus pneumoniae isolates of differing genetic backgrounds. Serotype 6C isolates are increasing in prevalence and becoming progressively more resistant to antibiotics. As a result, these strains are likely to become more important in the near future.

Mapping of AFLP markers linked to seed coat colour loci in Brassica juncea (L.) Czern.

Association mapping of the seed-coat colour with amplified fragment length polymorphism (AFLP) markers was carried out in 39 Brassica juncea lines. The lines had genetically diverse parentages and varied for seed-coat colour and other morphological characters. Eleven AFLP primer combinations were used to screen the 39 B. juncea lines, and a total of 335 polymorphic bands were detected. The bands were analysed for association with seed-coat colour using multiple regression analysis. This analysis revealed 15 markers associated with seed-coat colour, obtained with eight AFLP primer combinations. The marker E-ACA/M-CTG(350 )explained 69% of the variation in seed-coat colour. This marker along with markers E-AAC/M-CTC(235 )and E-AAC/M-CTA(250) explained 89% of the total variation. The 15 associated markers were validated for linkage with the seed-coat colour loci using a recombinant inbred line (RIL) mapping population. Bands were amplified with the eight AFLP primer combinations in 54 RIL progenies. Of the 15 associated markers, 11 mapped on two linkage groups. Eight markers were placed on linkage group 1 at a marker density of 6.0 cM, while the remaining three were mapped on linkage group 2 at a marker density of 3.6 cM. Marker E-ACA/M-CTG(350 )co-segregated with Gene1 controlling seed-coat colour; it was specific for yellow seed-coat colour and mapped to linkage group 1. Marker E-AAC/M-CTC(235) (AFLP8), which had been studied previously, was present on linkage group 2; it was specific for brown seed-coat colour. Since AFLP markers are not adapted for large-scale applications in plant breeding, it is important to convert these to sequence-characterised amplified region (SCAR) markers. Marker E-AAC/M-CTC(235) (AFLP8) had been previously converted into a SCAR. Work is in progress to convert the second of the linked markers, E-ACA/M-CTG(350), to a SCAR. The two linked AFLP markers converted to SCARs will be useful for developing yellow-seeded B. juncea lines by means of marker-assisted selection.

Screening interspecific hybrids of Populus (P. ciliata x maximowiczii) using AFLP markers.

Hybrids of Populus ciliata x maximowiczii are very vigorous and outperform both the parents in growth performance and yield. Genetic evaluation of 24 of these interspecific hybrids along with the two mother trees ( Populus ciliata), and five male-parent ( Populus maximowiczii) genotypes was carried out using the AFLP marker assay. Eight AFLP primer combinations detected 428 markers, of which 280 (66%) were polymorphic. Genetic relationships within the samples were evaluated by generating the similarity matrix based on Jaccard's coefficient. The phenetic dendrograms, as well as the PCO plots, separated the hybrids and the two parent species into three distinct clusters. The hybrids grouped closer to the P. ciliata (female parent) cluster as compared to the P. maximowiczii (male parent) cluster. The hybrid cluster contained internal groupings, which correlated to some extent with growth performance. The four best performing hybrids (42m1, 65m1, 23m2, Cm2-5-20/91) formed a distinct sub-cluster. Data from a single primer combination was sufficient for distinguishing the hybrids from the parents and assigning paternity. The hybrids showed 22 markers that were absent in P. ciliata but were monomorphically present in all the hybrids, suggesting outcrossing and common paternity. Further, these 22 markers were found in all the P. maximowiczii genotypes confirming it as the male parent. These male-specific markers can be converted to SCAR markers and used for rapid screening of the P.ciliata x maximowiczii hybrids. The primer combination E-AAC x M-CAA was identified as most suitable for ascertaining true hybridity. AFLP proves to be a useful tool for screening of P. ciliata x maximowiczii hybrids at the early stages of development.

Resorptive state and cell size influence intracellular pH regulation in rabbit osteoclasts cultured on collagen-hydroxyapatite films.

Diseases exhibiting excessive bone loss are often characterized by an increase in the size and number of osteoclasts in affected areas, suggesting that osteoclast size is associated with increased resorptive activity or efficiency. Because osteoclastic bone resorption depends on proton extrusion via a bafilomycin A1-sensitive vacuolar type H+ ATPase (V-ATPase), we investigated the relationship between osteoclast size and state of activity on the one hand, and proton-extruding mechanisms (bafilomycin A1-sensitive V-ATPase and amiloride-sensitive Na+/H+ exchange) on the other. In determining resorptive activities of individual osteoclasts, osteoclast-containing cell suspensions obtained from newborn rabbit long bones were cultured on apatite-collagen complex (ACC)-coated coverslips. Large osteoclasts resorbed 2.5 times more per cell than small osteoclasts, but the amount resorbed per nucleus was the same for the two categories. However, a much larger percentage of large osteoclasts was resorbing compared with small osteoclasts. To study pH regulatory mechanisms in individual large and small osteoclasts, the cells were loaded with the pH-sensitive indicator BCECF and analyzed by single-cell fluorescence. Small and large resorbing osteoclasts had significantly higher basal pH(i) than their nonresorbing counterparts. Also, small nonresorbing osteoclasts were insensitive to bafilomycin A1 addition or Na+ removal from the medium, large nonresorbing osteoclasts responded slightly, and all resorbing osteoclasts (small and large) responded strongly. Differences were also seen in the recovery from an acid load: both small and large nonresorbing osteoclasts were more sensitive to amiloride inhibition, while large resorbing cells were more sensitive to bafilomycin A1 inhibition. Small resorbing cells were inhibited equally by bafilomycin A1 and amiloride. These results clearly show that a greater proportion of large osteoclasts are active in resorption and that pH(i) regulation is associated with enhanced proton pump activity in actively resorbing osteoclasts. Thus, large and small osteoclasts differ in the proportion of cells that are resorbing, while pH regulatory mechanisms differ mainly between resorbing and nonresorbing cells.

Prophylactic blood patch performed prior to continuous epidural analgesia.

Two cases of inadvertent dural puncture in patients at high risk for dural puncture headache are presented. These patients were managed with an immediate epidural blood patch through a catheter placed at another level with placement of this catheter occurring immediately following the dural puncture. Following the blood patch, the patients were anesthetized with the epidural catheter being used for supplementation of the general anesthetic and for postoperative pain management. Neither patient required further management of a dural puncture headache nor did it appear that the blood interfered with the patients receiving adequate postoperative analgesia through the use of this epidural catheter.

Mutants of Streptococcus lactis subsp. diacetylactis lacking diacetyl reductase activity.

Three strains of Streptococcus lactis subsp. diacetylactis, namely DRC-1, DRC-2 and DRC-3 which produced diacetyl up to 120 h of incubation were exposed to the ultraviolet irradiation as well as N-methyl-N'-nitro-N-nitrosoguanidine (NTG) to isolate mutants lacking diacetyl reductase activity. UV irradiation did not produce any isolate completely devoid of diacetyl reductase activity, though, 99.5% loss in activity could be achieved. NTG treatment proved to be more effective and seven survivors exhibiting complete loss of diacetyl reductase activity were recovered. These altered characteristics were retained on repeated subculturing.