Treatment of bone loss in proximal femurs of postmenopausal osteoporotic women with AGN1 local osteo-enhancement procedure (LOEP) increases hip bone mineral density and hip strength: a long-term prospective cohort study.

This first-in-human study of AGN1 LOEP demonstrated that this minimally-invasive treatment durably increased aBMD in femurs of osteoporotic postmenopausal women. AGN1 resorption was coupled with new bone formation by 12 weeks and that new bone was maintained for at least 5-7 years resulting in substantially increased FEA-estimated femoral strength. INTRODUCTION: This first-in-human study evaluated feasibility, safety, and in vivo response to treating proximal femurs of postmenopausal osteoporotic women with a minimally-invasive local osteo-enhancement procedure (LOEP) to inject a resorbable triphasic osteoconductive implant material (AGN1). METHODS: This prospective cohort study enrolled 12 postmenopausal osteoporotic (femoral neck T-score ≤ - 2.5) women aged 56 to 89 years. AGN1 LOEP was performed on left femurs; right femurs were untreated controls. Subjects were followed-up for 5-7 years. Outcomes included adverse events, proximal femur areal bone mineral density (aBMD), AGN1 resorption, and replacement with bone by X-ray and CT, and finite element analysis (FEA) estimated hip strength. RESULTS: Baseline treated and control femoral neck aBMD was equivalent. Treated femoral neck aBMD increased by 68 ± 22%, 59 ± 24%, and 58 ± 27% over control at 12 and 24 weeks and 5-7 years, respectively (p < 0.001, all time points). Using conservative assumptions, FEA-estimated femoral strength increased by 41%, 37%, and 22% at 12 and 24 weeks and 5-7 years, respectively (p < 0.01, all time points). Qualitative analysis of X-ray and CT scans demonstrated that AGN1 resorption and replacement with bone was nearly complete by 24 weeks. By 5-7 years, AGN1 appeared to be fully resorbed and replaced with bone integrated with surrounding trabecular and cortical bone. No procedure- or device-related serious adverse events (SAEs) occurred. CONCLUSIONS: Treating femurs of postmenopausal osteoporotic women with AGN1 LOEP results in a rapid, durable increase in aBMD and femoral strength. These results support the use and further clinical study of this approach in osteoporotic patients at high risk of hip fracture.

Endocarditis caused by culture-negative organisms visible by Brown and Brenn staining: utility of PCR and DNA sequencing for diagnosis.

Two cases of culture-negative endocarditis with cocci seen in valve vegetations are presented. The organisms were identified by molecular analysis using broad-range PCR primers complementary to the 16S rRNA gene, sequencing, and database search using BLAST software. The results and utility of this method are discussed.

Evaluation of very low-dose subcutaneous vitamin K during postoperative warfarin therapy.

STUDY OBJECTIVE: To determine the effect of very low-dose subcutaneous vitamin K (SCVK) compared with withholding warfarin for above-target international normalized ratio (INR) values after joint surgery. DESIGN: Historical controlled study. SETTING: University hospital. SUBJECTS: One hundred thirty-nine patients beginning warfarin after total joint surgery. INTERVENTION: For a high INR, warfarin was either withheld or SCVK 100, 300, or 400 microg was administered, depending on INR value. MEASUREMENTS AND MAIN RESULTS: The primary outcome was change in INR from the day of intervention (day 1) to the next day (day 2). Adjusting for day 1 INR, the mean day 2 INR was 2.10 (95% confidence interval [CI] 1.86-2.33) after SCVK, compared with 2.73 (95% CI 2.50-2.96) in controls. This corresponded to declines of -0.72 and -0.08, respectively (p=0.001). CONCLUSION: In orthopedic patients starting warfarin therapy, very low-dose SCVK was more effective than withholding warfarin in reducing high INRs. Investigations in other populations and assessment of the effect of low-dose SCVK on postoperative bleeding are indicated.

Platelet PlA2 polymorphism enhances risk of neurocognitive decline after cardiopulmonary bypass. Multicenter Study of Perioperative Ischemia (McSPI) Research Group.

BACKGROUND: Neurocognitive decline, often produced by atherosclerotic plaque embolization, remains a frequent complication of cardiopulmonary bypass. Plaque fragments may initiate local thrombosis, which, in turn, aggravates the embolic insult. Prothrombotic genetic factors may exacerbate this process. We investigated whether the PlA2 polymorphism of platelet GPIIIa, a prothrombotic risk factor in other cardiovascular settings, is associated with early neurocognitive decline after cardiopulmonary bypass. METHODS: Neurocognitive changes were evaluated by the Mini-Mental State Examination administered preoperatively and on postoperative day 4 and the PlA genotype determined in 70 patients undergoing cardiopulmonary bypass. RESULTS: Forty-nine patients were PlA1/A1, and 21 were PlA1/A2 or PlA2/A2. Fifty-two patients (74%) demonstrated post-cardiopulmonary bypass neurocognitive decline, of which 34 were PlA1/A1 and 18 were PlA1/A2 or PlA2/A2 Multivariate analysis revealed that the PlA2 genotype and baseline Mini-Mental State Examination were significantly associated with greater neurocognitive decline (decreased Mini-Mental State Examination scores, p = 0.036 and 0.024, respectively). CONCLUSIONS: This study demonstrates a link between the PlA2 allele of platelet GPIIIa and more severe neurocognitive decline after cardiopulmonary bypass. Although the mechanism is unknown, it could represent exacerbation of platelet-dependent thrombotic processes associated with plaque embolism.

Rapid reconstitution of Epstein-Barr virus-specific T lymphocytes following allogeneic stem cell transplantation.

Epstein-Barr virus (EBV)-specific CD8 T lymphocytes are present at remarkably high frequencies in healthy EBV(+) individuals and provide protection from EBV-associated lymphoproliferative diseases. Allogeneic peripheral blood stem cell transplantation (allo-PBSCT) is a commonly used therapy in which T-cell surveillance for EBV is temporarily disrupted. Herein, human leukocyte antigen (HLA) class I tetramers were used to investigate the reestablishment of the EBV-specific CD8 T-cell repertoire in patients following allo-PBSCT. CD8(+) T cells specific for lytic and latent cycle-derived EBV peptides rapidly repopulate the periphery of matched sibling allo-PBSCT patients. The relative frequencies of T cells specific for different EBV peptides in transplantation recipients closely reflect those of their respective donors. Investigation of patients at monthly intervals following unmanipulated allo-PBSCT demonstrated that the frequency of EBV-specific T cells correlates with the number of EBV genome copies in the peripheral blood and that expansion of EBV-specific T-cell populations occurs even in the setting of immunosuppressive therapy. In contrast, patients undergoing T-cell-depleted or unrelated cord blood transplantation have undetectable EBV-specific T cells, even in the presence of Epstein-Barr viremia. The protective shield provided by EBV-specific CD8 T cells is rapidly established following unmanipulated matched sibling allo-PBSCT and demonstrates that HLA class I tetramers complexed with viral peptides can provide direct and rapid assessment of pathogen-specific immunity in this and other vulnerable patient populations. (Blood. 2000;96:2814-2821)

Pediatric AIDS-associated lymphocytic interstitial pneumonia and pulmonary arterio-occlusive disease: role of VCAM-1/VLA-4 adhesion pathway and human herpesviruses.

Because the mechanisms of lymphocyte accumulation in the lungs of children with AIDS-associated lymphocytic interstitial pneumonia (LIP) are unknown, we studied the relative contributions of known adhesion pathways in mediating lymphocyte adherence to endothelium and the potential role of human herpesviruses in the expansion of these lesions. LIP was characterized by lymphoid hyperplasia of the bronchus-associated lymphoid tissue (BALT) and infiltration of the pulmonary interstitium with CD8(+) T lymphocytes. In some individuals there was expansion of the alveolar septae with dense aggregates of B lymphocytes, many containing the Epstein-Barr viral (EBV) genome. Patients with concurrent EBV infection also demonstrated large-vessel arteriopathy characterized by thickening of the intimae with collagen and smooth muscle. Venular endothelium from the lung of children with LIP, but not uninflamed lung from other children with AIDS or lung from children with nonspecific pneumonitis, expressed high levels of vascular cell adhesion molecule-1 (VCAM-1) protein. In turn, inflammatory cells expressing very late activation antigen-4 (VLA-4), the leukocyte ligand for VCAM-1, were the predominant perivascular infiltrate associated with vessels expressing VCAM-1. Expression of other endothelial adhesion molecules, including intracellular adhesion molecule-1 and E-selectin, was not uniformly associated with LIP. Using a tissue adhesion assay combined with immunohistochemistry for VCAM-1, we show that CD8(+) T cell clones that express VLA-4 bind preferentially to pulmonary vessels in sites of LIP: vessels that expressed high levels of VCAM-1. When tissues and cells were pretreated with antibodies to VCAM-1 or VLA-4, respectively, adhesion was inhibited by >/=80%. Thus, infiltration of alveolar septae with CD8(+) T cells was highly correlative with VCAM-1/VLA-4 adhesive interactions, and focal expansion of B cells was coincidental to co-infection with EBV.

Reduced incidence of Epstein-Barr virus-associated posttransplant lymphoproliferative disorder using preemptive antiviral therapy.

BACKGROUND: Posttransplant lymphoproliferative disorder (PTLD) has been observed with increasing frequency consequent to the availability of more effective and potent immunosuppression. Prior work suggested that a peripheral blood monitoring strategy detecting peripheral B lymphoproliferation was effective in the early diagnosis of PTLD among 7 of 179 (3.9%) consecutive transplant recipients. Each of those seven patients received at least one course of antithymocyte globulin, Minnesota antilymphocyte globulin, or OKT3 before developing PTLD. METHODS: To determine whether antiviral prophylaxis might reduce the incidence of PTLD, a subsequent group of 198 consecutive recipients received either ganciclovir or acyclovir during antilymphocyte antibody administration. When the donor or recipient were cytomegalovirus-seropositive, ganciclovir was given; acyclovir was used when both were cytomegalovirus-seronegative. Baseline and protocol posttransplant cell surface profiles were obtained using immunofluorescence and flow cytometry to detect T cells, lymphocyte activation markers, and the CD19 B cell antigen. RESULTS: Demographic factors, including the incidence of recipients more than 50 years of age, non-Caucasians, previous transplantation, and diabetes mellitus, were similar in both groups. Additionally, the number of patients receiving antilymphocyte preparations was similar. However, only one patient (0.5%) from the latter group who received preemptive antiviral therapy developed PTLD. Although elevations in CD19+ B cells preceded clinical PTLD among each of the seven earlier patients, evidence of peripheral B cell proliferation was not demonstrated for the sole patient from the latter group, which suggests a possible effect of antiviral therapy. CONCLUSIONS: Prophylactic antiviral therapy may reduce the sensitivity of peripheral monitoring for B lymphoproliferation, but the dramatic reduction in PTLD incidence strongly supports its use among transplant recipients at risk.

Epstein-Barr virus suspension cell assay using in situ hybridization and flow cytometry.

We have developed a procedure for quantitative assay of Epstein-Barr virus (EBV)-infected cells in suspension in either latent or replicative phase using in situ hybridization and flow cytometry. The cells were hybridized with EBV-specific digoxigenin or biotin-labeled oligonucleotide probes, followed by binding to fluorescein-conjugated anti-digoxigenin or phycoerythrin-conjugated streptavidin, respectively. The cells hybridizing to the specific probes were quantitated by flow cytometry. A strong shift in fluorescence intensity (20-fold) was observed when the EBV-positive culture cells were hybridized with a specific EBER1 antisense probe. The sensitivity of the assay was at least one positive cell out of 9,000 beyond the normal control mean +/- 2 S.D. We performed two-color in situ hybridization/flow cytometry using probes to an EBV replication phase-specific mRNA and EBER1 on B95-8 cells in which a small portion (2-4%) of cells induce spontaneously into the replicative phase. In addition, we have developed a method for simultaneous analysis of the cell surface phenotype and EBV nucleic acid content in individual cells.

Cementless hemispheric acetabular components. A 4- to 8-year follow-up report.

A retrospective radiographic analysis was performed on 71 cementless acetabular components followed from 4 to 8 years after surgery. A Harris-Galante (Zimmer, Warsaw, IN) cup was used in 46 cases and an Optifix (Smith & Nephew Richards, Memphis, TN) cup was used in 25 cases. There were 66 primary and 5 revision cases. Nonprogressive radiolucencies were common in both types of cups (64% of Optifix, 70% of Harris-Galante). Radiolucencies were almost exclusively less than or equal to 1 mm in width and were most common in zones 1 and 3. Ten cups had continuous but nonprogressive radiolucencies, none greater than 1 mm in all three zones. No radiolucent lines wider than 2 mm were seen in any case. Four cups had progressive radiolucency that stabilized. Radiolucency around fixation screws was seen in one case, and demonstrated stable ingrowth at revision surgery. No cases of osteolysis, screw breakage, migration, or loss of fixation surface occurred. A single case of a broken locking mechanism of a Harris-Galante cup 2 years after liner exchange is reported.

Posterior cruciate ligament strain biomechanics in total knee arthroplasty.

To evaluate the role of the posterior cruciate ligament in total knee arthroplasty, 8 normal whole-leg cadaveric legs were studied. Strain patterns of the posterior cruciate ligament were measured during active and passive knee flexion extension. Total knee arthroplasty was performed, and posterior cruciate ligament strain was measured again. The standard tibial insert was removed, and 3 additional tibial trays were used: flat, 10 degrees sloped, and 15 degrees sloped. The posterior cruciate ligament strain was recorded for each tibial insert. Total knee arthroplasty with a standard insert produced a large range of posterior cruciate ligament strain values (+/- 6%). Of the 8 specimens, 3 produced excessively taut posterior cruciate ligament strain, 3 were slack, and 2 returned strain to the baseline strain value. Changing the surface contour and slope of the tibial inserts did not produce any consistent change in the posterior cruciate ligament strain pattern. The ability of total knee arthroplasty to reproduce normal posterior cruciate ligament strain behavior has not been demonstrated.

Tibial plateau coverage in total knee arthroplasty.

Optimizing coverage of the resected tibial plateau is an important consideration in total knee arthroplasty. The tibial coverage of eight different tibial tray designs was examined in 35 resected tibial specimens. When no component overlap was permitted, the average tibial coverage of the different designs ranged from 76.4% to 80.8%. When the component was allowed slight overlap, a "surgical fit," the average coverage ranged from 78.5% to 85.5%. As a group, the six symmetric designs provided more coverage than the two asymmetric designs (p < 0.05). Areas of poor coverage were identified, and different tray geometries were compared. Asymmetric tibial components are thought to be more anatomic in nature and therefore cover the tibial plateau surface more completely than symmetric components. The results of this study do not support this, and in fact demonstrate that symmetric components can provide more coverage when compared with two asymmetric component designs.

Cementless hemispheric acetabular components. 2-4-year results.

A retrospective clinical and radiographic analysis was performed on 99 patients (106 hips) undergoing total hip arthroplasty with noncemented Harris-Galante (Zimmer, Warsaw, IN) or Optifix (Smith Nephew Richards, Memphis, TN) acetabular components. There were 94 primary and 12 revision procedures with a minimum follow-up period of 24 months (range, 24-52 months). The cups were evaluated for evidence of vertical and horizontal migration, as well as the presence of radiolucencies at the bone-implant interface. Age, sex, component inclination and medialization, cup coverage, and number of fixation screws used were examined to determine the influence of these factors on the incidence of cup migration or radiolucent line formation. A radiolucent line was present in at least one zone in 60% of the Harris-Galante and 45% of the Optifix cups. Progressive radiolucent lines were noted in two of the Optifix and three of the Harris-Galante components. Two Harris-Galante cups (1.9%) were revised. There were no Optifix cup migrations or evidence of instability in the Optifix or remaining Harris-Galante cups. Although there was a trend toward an increased incidence of radiolucencies in those cups lacking complete coverage, no statistically significant radiographic predictors for failure or impending failure could be determined from this short-term follow-up study. With the exception of the two revisions, all acetabular components performed well clinically over the study period.

Failure of the polyethylene liner leading to notching of the femoral component in bipolar prostheses.

Polyethylene wear is an increasingly recognized problem in joint replacement surgery. Three cases of polyethylene wear in bipolar hip prostheses are presented. Owing to failure of the polyethylene liner, impingement of the metal shell produced notching of the femoral component.

Upstream basal promoter element important for exclusive RNA polymerase III transcription of the EBER 2 gene.

The Epstein-Barr virus-encoded small RNA (EBER) genes are transcribed by RNA polymerase III, but their transcription unit appears to contain both class II and class III promoter elements. One of these promoter element, a TATA-like box which we call the EBER TATA box, or ETAB, is located in a position typical for a class II TATA box but contains G/C residues in the normal T/A motif and a conserved thymidine doublet. Experiments using chloramphenicol acetyltransferase constructs and mutations in the TATA box of the adenovirus major late promoter showed that the ETAB promoter element does not substitute for a class II TATA box. However, when the ETAB promoter element sequence was changed to a class II TATA box consensus sequence, the EBER 2 gene was transcribed in vitro by both RNA polymerases II and III. From these results, we conclude that the ETAB promoter element is important for the exclusive transcription of the EBER 2 gene by RNA polymerase III.

The effect of functional knee-braces on strain on the anterior cruciate ligament in vivo.

Functional knee-braces are widely used to protect injured or reconstructed anterior cruciate ligaments, despite the fact that few scientific data support their efficacy. We studied seven functional braces, representative of both the typical custom-fit and off-the-shelf designs. The braces were tested on subjects who had a normal anterior cruciate ligament and were scheduled for arthroscopic meniscectomy or exploration of the knee under local anesthesia. After the operative procedure, a Hall-effect strain-transducer was applied to the anterior cruciate ligament. Under low anterior shear loads, two braces provided some protective strain-shielding effect compared with no brace, but this strain-shielding effect did not occur at the higher anterior shear loads expected during the high-stress activities common to athletic events. The DonJoy, Townsend, C.Ti., and Lenox Hill braces demonstrated a strain-shielding effect on the anterior cruciate ligament with an internal torque of five newton-meters applied to the tibia. None of the braces had any effect on strain on the anterior cruciate ligament during active range of motion of the knee from 10 to 120 degrees or during isometric contraction of the quadriceps. Wearing of a brace did not produce an increase in the value for strain on the anterior cruciate ligament. For the activities that were evaluated in this study, none of the braces produced adverse effects on the anterior cruciate ligament, and there were no significant differences in the strain on the anterior cruciate ligament between the use of a custom-fit or an off-the-shelf brace design. There were no apparent advantages of the more expensive custom-made braces compared with the off-the-shelf designs.

Clinical versus instrumented knee testing on autopsy specimens.

The accuracy, reliability, and reproducibility of the Genucom Knee Analysis System and Knee Signature System (KSS) for anteroposterior knee laxity evaluations were compared. The devices detected the same relative change in laxity between normal and anterior cruciate ligament-deficient specimens during Lachman and drawer testing; however, the absolute values differed. In a clinical study, two examiners performed three independent Lachman and drawer tests using the Genucom, KSS, and a subjective clinical examination on ten patients. The Genucom demonstrated interexaminer differences during the Lachman test. The clinical examination proved to be more reliable than either instrumented device for both Lachman and drawer testing. The intraexaminer variability of the Genucom and KSS was large, with the average 95% confidence limits about the mean for the Genucom and KSS equal to +/- 4.2 and +/- 2.8 mm, respectively. These findings question the accuracy, reliability, and reproducibility of the instrumented methods.

The measurement of anterior cruciate ligament strain in vivo.

This article describes the use of the Hall Effect strain transducer (HEST) in a new arthroscopic technique to study the normal anterior cruciate ligament (ACL) in-vivo. Study participants were patient volunteers with normal ACLs undergoing diagnostic arthroscopic or meniscal surgery under local anaesthesia. The HEST was implanted into the Anterior Medial Band (AMB) of the ACL. Anterior shear loading of the tibia in relation to the fixed femur at 30 degrees of knee flexion (Lachman test), produced significantly greater strain values in comparison to anterior shear loading at 90 degrees (Anterior Drawer test). During isometric quadriceps contraction a significant increase in AMB strain was measured with the knee flexed to 30 degrees, while no significant change was measured at 90 degrees. For quadriceps contraction there were significantly higher values of AMB strain measured at 30 degrees of knee flexion in comparison to that observed at 90 degrees. For active range of motion (AROM) the AMB was strained between 10 degrees and 48 degrees, and unstrained between 48 degrees and 110 degrees. During passive range of motion (PROM) the AMB remained unstrained until the joint was brought into extension. There were significant differences in strain values found between AROM and PROM at the flexion angles 10 degrees, 20 degrees, 30 degrees and 40 degrees, while between 50 degrees and 110 degrees there were no significant differences. These results confirm previous studies that the Lachman test is a superior technique in comparison to the classic anterior drawer test for evaluating the AMB. They suggest that isometric quadriceps activity at 90 degrees of knee flexion can be prescribed for rehabilitation immediately after ACL reconstruction. These data indicate that AROM (between the limits of 50 degrees and 110 degrees) and PROM may also be performed with minimal risk of strain to a reconstructive replacement. The PROM data may also serve as an important standard for the reconstruction of the ACL.

Longitudinal crack propagation in bone around femoral prosthesis.

Intraoperative femoral fracture is a well-recognized technical complication of cementless total hip arthroplasty. This study was designed to establish an in vitro model for initiation and propagation of fractures of the proximal femur in cementless THA and to assess the effect of fracture fixation in the form of cerclage wiring and drilling a hole in the distal extent of the fracture line. Fourteen human anatomic femur specimens were studied. Longitudinal cracks were made and propagation was performed on a materials testing system machine. A drill hole at the tip of a longitudinal crack does not prevent crack propagation. However, cerclage wiring has a statistically significant effect (p less than 0.025) on the ability of the fractured femur to withstand increased load.