Large multi-chromosomal duplications encompass many members of the olfactory receptor gene family in the human genome

The human genome contains thousands of genes that encode a diverse repertoire of odorant receptors (ORs). We report here on the identification and chromosomal localization of 74 OR-containing genomic clones. Using fluorescence in situ hybridization (FISH), we demonstrate a striking homology among a set of approximately 20 OR locations, illustrating a history of duplications that have distributed OR sequences across the genome. Half of the OR-containing BACs cloned from total genomic DNA and 86% of cosmids derived from chromosome 3 cross-hybridize to a subset of these locations, many to 17 of them. These paralogous regions are distributed on 13 chromosomes, and eight lie in terminal bands. By analyzing clones from an approximately 250 kb clone-walk across one of these sites (3p13), we show that the homology among these sites is extensive (>150 kb) and encompasses both OR genes and intergenic genomic sequences. The FISH signals appear significantly larger at some sites than at the native location, indicating that portions of some duplicons have undergone local amplification/attrition. More restricted duplications involving pairs of other genomic locations are detected with 12% of the OR-BACs. Only a small subset of OR locations is sufficiently diverged from the others that clones derived from them behave as single-copy FISH probes. We estimate that duplications encompassing members of the OR gene family account for >0.1% of the human genome. A comparison of FISH signals at orthologous locations in other primates indicates that a portion of this OR 'subgenome' has been in flux during the divergence of primates, possibly as a mechanism for evolving the repertoire of olfactory receptors.     

Comparison of closed-suction drainage and no drainage after primary total knee arthroplasty

One hundred thirty-six primary total knee arthroplasty patients were randomized for the use of closed-suction, nonreinfusable wound drains. Blood loss was identical in the drained and undrained groups. Forty percent of undrained wounds compared with 0% of drained wounds required dressing reinforcement. Sixty-nine percent of undrained wounds compared with 39% of drained wounds developed ecchymosis, measuring 92 cm2 in the undrained group and 28 cm2 in the drained group. This study concludes that a simple wound drain effectively minimizes the undesirable accumulation of blood in the surrounding soft tissues and the postoperative wound dressing after total knee arthroplasty.     

Treatment of complications in primary cementless total hip arthroplasty

Five hundred eight consecutive cases (481 patients) treated with the extensively porous coated Anatomic Medullary Locking prosthesis were followed for an average of 9 years (range, 5-14 years). Thirty-one (6%) hips were lost to followup and 33 (7%) hips had complications that required revision surgery. The indications for revision were symptomatic stem loosening (six cases), symptomatic cup loosening (five cases), asymptomatic periarticular osteolysis (seven cases); trochanteric fracture through an osteolytic cyst (four cases), polyethylene fracture (five cases), sepsis (one case), and heterotopic ossification (one case). The surgical treatment of these complications is described. After these revisions, 11 (33%) cases had additional complications, most commonly a dislocation. Four required a second revision. Questionnaires and physical examinations were used to compare the outcome of the cases requiring revision with the outcome of those that did not. There were no differences in patient satisfaction between cases requiring revision surgery and those that did not (97% and 95% patient satisfaction, respectively). Function was also similar between the two groups, with 93% reporting increased function in each group.     

The Coventry Award. Modular tibial insert micromotion. A concern with contemporary knee implants

This study investigates micromotion between modular tibial components, one of the causes of wear on the undersurface of polyethylene inserts. The authors measured motion at the interface of nine contemporary total knee implant designs by mechanically testing the implants in a servohydraulic testing machine. Anteroposterior and mediolateral motion between the tibial insert and baseplate were measured with an extensometer placed across the interface. These tests revealed that in all implants analyzed, sufficient motion occurred to create fretting at the modular interface. Although the testing configuration in this study was not a stimulation of in situ loading patterns, the authors observed hundreds of microns of motion even under a 100 N load and variability between implants of the same design, showing that there is room for improvement among locking mechanisms in modular total knee implants.     

Cementless acetabular components: optimums and outcomes

The first known case of an intravascular catheter-related primary cutaneous mucormycosis in a heart transplant patient is reported. The patient had corticosteroid-induced hyperglycemia and experienced an acute tissue rejection episode. A necrotic lesion appeared around the insertion site of a peripheral venous catheter. A biopsy revealed typical mucorales hyphae. The lesion continued to spread during the following 24 hours and necessitated amputation of the forearm. The organism was identified as a Mucor species.     

Porous-coated total hip replacement

Fifteen years of clinical experience with porous-coated prostheses demonstrated the durability of this type of fixation. This experience was documented by clinical follow-up study of the 393 cases treated by the senior author before 1985. Only six of these femoral components have been revised: three for loosening, two for stem breakage, and one for infection. Thus, the revision rate for the porous-coated stems was 1.5%. Porous-coated acetabular components were used in 227 of the arthroplasties. Five of these porous-coated cups have been revised: four for malposition leading to dislocation and one for late loosening secondary to osteolysis. Thus, the revision rate for these porous-coated acetabular components was 2.2%. Twenty bipolar and 146 cemented acetabular components were used in the remaining 166 cases treated before 1985. Eleven (7.5%) of the cemented acetabular components were revised. Revisions of the porous-coated components were rare in the first ten postoperative years. The clinical data were supplemented with analysis of postmortem specimens from 15 patients. Mechanical testing of the femoral specimens showed the relative micromotion at the porous surface to be exceptionally small (less than 40 microns). Seven of these postmortem retrievals involved cases with unilateral arthroplasties. In these cases, the contralateral normal femur also was removed, and a prosthesis identical to that in the in vivo implanted side was inserted to simulate the immediate postoperative condition. Dual-energy X-ray absorptiometry (DEXA) of the seven paired femora demonstrated that bone remodeling can be expected to produce a 5%-52% loss of periprosthetic bone mineral content, with the greatest loss occurring in the more osteoporotic patients.(ABSTRACT TRUNCATED AT 250 WORDS)     

The influence of clinical factors on periprosthetic bone remodeling

The femurs of 11 patients with well-functioning unilateral hip replacements were retrieved at autopsy and analyzed for periprosthetic bone remodeling by dual energy xray absorptiometry. Each case involved a femur with a porouscoated endoprosthesis; the endoprosthesis remained implanted for an average of 5.9 years. In the contralateral femur, a matching prosthesis was implanted in vitro, to serve as a control for comparisons. There was an average 22.6% decrease in bone mineral content in the in vivo implanted femur (range, 5.4%-47.4%). Females experienced an average bone loss of 31.2%, which was significantly higher than the 12.3% average loss in males. Longitudinal analysis revealed an average decrease in bone mineral content of 42.1% proximally, 23% in the midsection, and 5.5% distally. Percent decreases in total bone mineral content were correlated with the following clinical variables: weight, age, implant diameter, duration of implantation, and contralateral femoral bone mineral content. Only the bone mineral content of the contralateral femur had a strong predictive value. Bone loss was greater in femurs with low bone mineral content than in those with high bone mineral content. Weight, age, implant diameter, and duration of implantation were not correlated with bone loss.     

Diarylsulfonamides as selective, non-peptidic thrombin inhibitors

Based on the structures of aminopyridine thrombin inhibitors (1), a series of aminoalkyl- and guanidinoalkyl-substituted diarylsulfonamides were prepared. The most potent derivative, N-[3-(4-guanidinobutoxy)-5-methyl-phenyl]-benzenesulfonamide (6c) had Ki = 0.18 microM for thrombin and did not inhibit trypsin, plasmin, or factor Xa. Comparison of the X-ray structures of the thrombin/1b and the thrombin/6c complexes revealed important aspects which govern the binding of such diarylsulfonamides to thrombin.     

New enzyme lineages by subdomain shuffling

Protein functions have evolved in part via domain recombination events. Such events, for example, recombine structurally independent functional domains and shuffle targeting, regulatory, and/or catalytic functions. Domain recombination, however, can generate new functions, as implied by the observation of catalytic sites at interfaces of distinct folding domains. If useful to an evolving organism, such initially rudimentary functions would likely acquire greater efficiency and diversity, whereas the initially distinct folding domains would likely develop into single functional domains. This represents the probable evolution of the S1 serine protease family, whose two homologous beta-barrel subdomains assemble to form the binding sites and the catalytic machinery. Among S1 family members, the contact interface and catalytic residues are highly conserved whereas surrounding surfaces are highly variable. This observation suggests a new strategy to engineer viable proteins with novel properties, by swapping folding subdomains chosen from among protein family members. Such hybrid proteins would retain properties conserved throughout the family, including folding stability as single domain proteins, while providing new surfaces amenable to directed evolution or engineering of specific new properties. We show here that recombining the N-terminal subdomain from coagulation factor X with the C-terminal subdomain from trypsin creates a potent enzyme (fXYa) with novel properties, in particular a broad substrate specificity. As shown by the 2.15-A crystal structure, plasticity at the hydrophobic subdomain interface maintains activity, while surface loops are displaced compared with the parent subdomains. fXYa thus represents a new serine proteinase lineage with hybrid fX, trypsin, and novel properties.