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Biomedical subjects

J J Jacobs

Publications and source records attributed to J J Jacobs.

At least 55 records · Page 3Linked to original sources

Detection of varicella-zoster virus and herpes simplex virus by the polymerase chain reaction with degenerate primers.

Varicella-zoster virus (VZV) and herpes simplex virus (HSV) are human pathogens of significance involved in multiple diseases with either typical or atypical clinical features. In neonates and immunocompromised patients these alphaherpesviruses may cause life-threatening diseases such as encephalitis. Detection of VZV by virus culture is difficult. Polymerase chain reaction (PCR) is quicker and more sensitive and applicable in most clinical microbiological laboratories. Using degenerate primers, glycoprotein B (gB) DNA was amplified from all alphaherpesvirus field strains present in clinical samples. The amplification of gB allowed virus typing of VZV, HSV-1 and HSV-2 using restriction enzyme digestion of the PCR products. Degenerate primers can replace conventional primers in diagnostic PCR without loss of sensitivity and specificity.

Base Sequence↗

Posttranscriptional gene silencing of gn1 in tobacco triggers accumulation of truncated gn1-derived RNA species.

Posttranscriptional silencing of basic beta-1,3-glucanase genes in the tobacco line T17 is manifested by reduced transcript levels of the gn1 transgene and homologous, endogenous basic beta-1,3-glucanase genes. An RNA ligation-mediated rapid amplification of cDNA ends (RLM-RACE) technique was used to compare the 3' termini of gn1 RNAs present in expressing (hemizygous and young homozygous) and silenced (mature homozygous) T17 plants. Full-length, polyadenylated gn1 transcripts primarily accumulated in expressing plants, whereas in silenced T17 plants, mainly 3'-truncated, nonpolyadenylated gn1 RNAs were detected. The relative abundance of these 3'-truncated gn1 RNA species gradually increased during the establishment of silencing in homozygous T17 plants. Similar 3'-truncated, nonpolyadenylated gn1 RNA products were observed in an independent case of beta-1,3-glucanase posttranscriptional gene silencing. This suggests that these 3'-truncated gn1 RNAs are a general feature of tobacco plants showing posttranscriptional silencing of the gn1 transgene.

Base Sequence↗

Impaction allografting with cement for revision of the femoral component. A minimum four-year follow-up study with use of a precoated femoral stem.

BACKGROUND: Cancellous impaction allografting with cement for revision of the femoral component has conventionally been performed with a polished, tapered implant, which was designed to allow subsidence of the component. However, subsidence has been associated with pain in the thigh, dislocation of the hip, and revision of the component. This prospective study tested the hypothesis that good clinical results can be achieved--without subsidence of the component--with use of impaction allografting and a precoated, collared, straight stem for difficult femoral revisions. METHODS: Twenty-nine patients had revision of the femoral component with use of impaction allografting with cement and a Harris Precoat stem. Impaction allografting was performed when loss of metaphyseal and diaphyseal bone precluded revision with more straightforward techniques or when reconstitution of bone was considered a specific goal of the reconstruction (as was sometimes the case with revision of the component in younger patients). The patients were followed prospectively and were evaluated with use of the Harris hip score and serial radiographs. The patients were followed for a minimum of four years (mean, sixty-three months), except for four who died. RESULTS: Four patients died before the minimum four-year follow-up period had elapsed; all four had the prosthesis in place at the time of death. The Harris hip scores improved from a preoperative mean of 54 points (poor) (range, 21 to 91 points) to a mean of 87 points (good) (range, 41 to 100 points) at the time of the most recent follow-up. Kaplan-Meier survivorship analysis, with aseptic loosening as the end point, was 92 percent (95 percent confidence interval, 82 to 100 percent) at six years; one additional hip failed because of a hematogenous infection at seventy-three months postoperatively, for an overall failure rate of 12 percent (three of twenty-five patients) at the time of the most recent follow-up. Two hips needed a repeat revision; one was revised because of subsidence of the stem with recurrent osteolysis and the other, in a patient who had hemodialysis, because of late sepsis. A third femoral component subsided and failed but was not revised. Radiographic evidence of bone-stock reconstitution was observed in six (29 percent) of the twenty-one patients for whom radiographs were available. As in other series of patients managed with impaction allografting, the complication rate was high; excluding the revisions, three reoperations were performed, and six patients had either intraoperative femoral fracture or perforation necessitating cerclage wiring or cortical strut allografting and cerclage wiring at the time of the procedure. There were six nonunions in eighteen patients who had been operated on with a transtrochanteric approach. CONCLUSIONS: Difficult revisions of the femoral component with use of impaction allografting and a precoated stem provided satisfactory clinical and radiographic results at the time of intermediate-term follow-up. However, the high rate of complications in our series led us to refine our indications for the procedure.

Adult↗

The fate of stable cemented acetabular components retained during revision of a femoral component of a total hip arthroplasty.

BACKGROUND: The decision as to whether to revise or retain a well fixed cemented acetabular component during revision of a femoral component is especially difficult; the rate of loosening of cemented acetabular components is high, whereas that of porous-coated acetabular components inserted during revision is low. However, removal of a well fixed cemented acetabular component can result in increased operative morbidity and cost and in loss of acetabular bone. Data that can be used to predict the long-term survival of retained well fixed cemented acetabular components are therefore needed. METHODS: We studied the five to thirteen-year clinical and radiographic results in a group of twenty-six consecutive patients in whom a well fixed cemented acetabular component had been retained during revision of a femoral component. Typical demographic data on the patients and information about the components were recorded, and the cemented acetabular components were graded as A through F, according to the system of Ranawat et al., at the time of the femoral revision. The average duration of follow-up was 8.4 years (range, 5.0 to 12.7 years). No patient was lost to follow-up. RESULTS: Four acetabular components (15 percent) had progressive radiolucency (at forty-eight, forty-eight, fifty-nine, and seventy-five months after the femoral revision) and were considered radiographically loose despite not being associated with symptoms. All four components were graded as either E or F at the time that they were retained during the femoral revision; radiographic loosening was significantly related to these two grades (p < 0.01). No acetabular component with a grade of A, B, C, or D loosened. The components that loosened had been in vivo for a relatively shorter, as opposed to longer, duration before the femoral revision compared with the components that did not loosen (p < 0.05). CONCLUSIONS: Retention of the well fixed cemented acetabular components was associated with good clinical results but with a 15 percent rate of loosening. Revision of a cemented acetabular component solely on the basis of the duration that it was in vivo or whether a previous revision had been done does not appear to be warranted. Our findings suggest that acetabular components with a grade of A, B, C, or D at the time of a femoral revision may be retained, as these components continued to function at the time of the five to thirteen-year follow-up in the current study.

Acetabulum↗

Unicompartmental knee arthroplasty. Clinical experience at 6- to 10-year followup.

Sixty-two consecutive cemented modular unicompartmental knee arthroplasties in 51 patients were studied prospectively. At surgery, the other compartments had at most Grade 2 chondromalacia. The average age of the patients at arthroplasty was 68 years (range, 51-84 years). One patient was lost to followup and 10 died with less than 6 years followup. The average followup of the remaining 51 knees was 7.5 years (range, 6-10 years). The preoperative Hospital for Special Surgery knee score of 55 points (range, 30-79 points) improved to 92 points (range, 60-100 points) at followup; 78% (40 knees) had excellent and 20% (10 knees) had good results. The mean range of motion at followup was 120 degrees with 26 knees (51%) having range of motion greater than 120 degrees. One patient underwent revision surgery for retained cement, one patient underwent knee manipulation, and one patient underwent revision surgery at 7 years for opposite compartment degeneration and pain. Radiographically, 26 knees (51%) had at least one partial radiolucency. There were no complete femoral radiolucencies, but there were three complete tibial radiolucencies, all less than 2 mm. No component was loose as seen on radiographs. At final followup, five of the opposite compartments (10%) and three of the patellofemoral joints (6%) had some progressive radiographic joint space loss; this was less than a 25% loss in all but one knee component that was revised. At 6- to 10-years followup, cemented unicompartmental knee arthroplasty yielded excellent clinical and radiographic results. The 10-year survival using radiographic loosening or revision as the end point was 98%. Using stringent selection criteria, unicompartmental knee replacement can yield excellent results and represents a superb alternative to total knee replacement.

Aged↗

Metal release and excretion from cementless titanium alloy total knee replacements.

Concentrations of titanium, aluminum, and vanadium were measured in the serum and urine of patients with titanium alloy cementless primary total knee arthroplasty components. Patients were categorized in one of five groups. In Group 1, the patellar and tibial articulating surfaces were made of carbon fiber reinforced ultrahigh molecular weight polyethylene. In Group 2, the patellar and tibial surfaces were made of ultrahigh molecular weight polyethylene. In Group 3, the femoral titanium alloy articulating surface was nitrogen ion implanted with ultrahigh molecular weight polyethylene patellar and tibial articulating surfaces. Patients in Group 4 had failed patellar components, and Group 5 was comprised of age and gender matched control subjects without implants. Serum concentrations of titanium were approximately 50 times greater in patients with failed patellar components (Group 4) and approximately 10 times greater in patients with carbon fiber reinforced polyethylene bearing surfaces (Group 1) when compared with Groups 2 and 3 and the control subjects (Group 5). For aluminum and vanadium, no detectable differences were observed among any of the groups. In addition, analysis of 24-hour urine samples showed no significant differences in titanium, aluminum, or vanadium concentrations among any of the groups. Elevated serum titanium levels may serve as a marker of patellar component failure or accelerated femoral component wear in total knee replacements with titanium alloy bearings. The toxicologic ramifications of these findings are unknown.

Aged↗

Silencing of beta-1,3-glucanase genes in tobacco correlates with an increased abundance of RNA degradation intermediates.

Post-transcriptional gene silencing of beta-1,3 glucanase genes in the transgenic tobacco line T17 is characterised by an increased turnover and, as a consequence, reduced levels of gn1 transgene and endogenous beta-1,3 glucanase mRNAs. Here, additional gn1 RNAs, both larger and smaller than the full-length messenger, are shown to accumulate in silenced plants of the transgenic tobacco line T17. The longer-than-full-length gn1 RNAs are the result of cryptic processing of the gn1 messenger. The small gn1 RNAs in silenced plants correspond to distal and proximal parts of the mature gn1 messenger. The proximal RNA products are intact at their 5' extremity, but terminate at different positions at the 3'-end. The distal RNA products contain a poly(A) tail and are truncated to various positions at the 5'-end. These observations indicate that degradation of the mature gn1 transcript does not start at the 5'- or 3'-end, but rather are consistent with degradation of the gn1 transcript starting with an endonucleolytic cleavage followed by internal exonuclease digestion. Importantly, the truncated products are more abundant in silenced plants than in expressing plants. This suggests, together with the previously reported silencing-related increased gn1 mRNA turnover and the similar rates of gn1 transcription in silenced and expressing T17 plants, that the predominant decay route for the gn1 transcripts differs between silenced and expressing conditions.

Chromosome Mapping↗

Progressive polyarthritis induced in BALB/c mice by aggrecan from normal and osteoarthritic human cartilage.

OBJECTIVE: To find an "unlimited" source of antigenic material (aggrecan) for arthritis induction in BALB/c mice; to analyze the specificities of immune reactions to aggrecan and type II collagen in 2 arthritis-susceptible murine strains, BALB/c mice for proteoglycan (aggrecan)-induced arthritis and DBA/1j mice for collagen-induced arthritis; to compare the histopathologic features of arthritis induced by purified aggrecans or total extracts of osteoarthritic (OA) cartilage; and to determine arthritis susceptibility in various BALB/c colonies. METHODS: Aggrecans from total extracts of human fetal, normal adult, OA, and rheumatoid cartilage samples and from osteophytes were isolated, purified by gradient centrifugation, deglycosylated, characterized, and tested for arthritis induction. Purified type II collagen and salt-soluble collagens from OA cartilage were denatured, stromelysin treated, and used for immunization and arthritis induction in arthritis-susceptible (DBA/1j and BALB/c) murine strains. RESULTS: Chondrocytes from OA cartilage synthesize predominantly fetal-type aggrecan, which is the most efficient antigenic material for arthritis induction in BALB/c mice. The critical autoimmune/arthritogenic T cell epitopes of aggrecan are located in the G1 domain. Although most of the aggrecan molecules are heavily degraded and lost from OA cartilage, the G1 domain-containing fragments accumulate in OA cartilage. The amount of G1-containing fragments is approximately twice as much in OA than in normal adult articular cartilage, and the arthritogenic epitope(s) remains intact in G1-containing fragments retained in cartilage. Thus, total extracts of OA cartilage (without additional purification), if deglycosylated appropriately, can be used as arthritogenic material in BALB/c mice. CONCLUSION: Predominantly G1 domain-containing fragments of aggrecan accumulate in OA cartilage, and these are the fragments which induce arthritis in BALB/c mice. Arthritis induction is highly specific for aggrecan epitopes and dictated by the genetic background of the BALB/c strain.

Adult↗

Specific expression in mouse mesoderm- and neural crest-derived tissues of a human PDGFRA promoter/lacZ transgene.

The platelet-derived growth factor alpha-receptor (PDGFR-alpha) displays a lineage-specific expression pattern in the mouse embryo and is required for normal development of mesoderm and cephalic neural crest derivatives. The purpose of the present study was to demonstrate the in vivo promoter function of genomic DNA fragments representing the 5'-flanking part of the human PDGFRA gene. 2.2, 0.9 and 0.4 kb PDGFRA promoter fragments, ligated to a lacZ reporter gene, were microinjected into fertilized mouse eggs and transgenic mouse lines were established. The expression patterns were basically similar in the 2.2 and 0.9 kb lines and overlapped grossly the endogenous Pdgfra gene expression pattern. The transgenic line with the highest expression level was chosen for detailed analysis. Expression was, as expected, mainly confined to tissues of mesodermal and neural crest origin. No expression was found in epithelial tissues of endo- or ectodermal origin. The promoter fragments were also active in neuroepithelium and in certain neuronal cell types that did not faithfully express PDGFR-alpha mRNA, while they failed to specify reporter expression in PDGFR-alpha expressing O-2A progenitor cells and other glial elements of the central nervous system. Thus, the isolated human PDGFRA promoter contains most but not all of the regulatory elements that are necessary to establish tissue specific gene expression during development.

Animals↗

Malrotation causing patellofemoral complications after total knee arthroplasty.

Thirty patients with isolated patellofemoral complications after total knee arthroplasty were compared with 20 patients with well functioning total knee replacements without patellofemoral complications. The epicondylar axis and tibial tubercle were used as references on computed tomography scans to measure quantitatively rotational alignment of the femoral and tibial components. The group with patellofemoral complications had excessive combined (tibial plus femoral) internal component rotation. This excessive combined internal rotation was directly proportional to the severity of the patellofemoral complication. Small amounts of combined internal rotation (1 degree-4 degrees) correlated with lateral tracking and patellar tilting. Moderate combined internal rotation (3 degrees-8 degrees) correlated with patellar subluxation. Large amounts of combined internal rotational (7 degrees-17 degrees) correlated with early patellar dislocation or late patellar prosthesis failure. The control group was in combined external rotation (10 degrees-0 degree). The direct correlation of combined (femoral and tibial) internal component rotation to the severity of the patellofemoral complication suggests that internal component rotation may be the predominant cause of patellofemoral complications in patients with normal axial alignment. The epicondylar axis and tibial tubercle are reproducible landmarks which are visible on computed tomography scans and can be used intraoperatively. Using this computed tomography study can determine wether rotational malalignment is present and thus, whether revision of one or both components may be indicated.

Arthroplasty, Replacement, Knee↗

Metal release in patients who have had a primary total hip arthroplasty. A prospective, controlled, longitudinal study.

UNLABELLED: There is an increasing recognition that, in the long term, total joint replacement may be associated with adverse local and remote tissue responses that are mediated by the degradation products of prosthetic materials. Particular interest has centered on the metal-degradation products of total joint replacements because of the known toxicities of the metal elements that make up the alloys used in the implants. We measured the concentrations of titanium, aluminum, cobalt, and chromium in the serum and the concentration of chromium in the urine of seventy-five patients during a three-year prospective, longitudinal study. Twenty patients had had a so-called hybrid total hip replacement (insertion of a modular cobalt-alloy femoral stem and head with cement and a titanium acetabular cup without cement), fifteen had had insertion of an extensively porous-coated cobalt-alloy stem with a cobalt-alloy head and a titanium-alloy socket without cement, and twenty had had insertion of a proximally porous-coated titanium-alloy stem with a cobalt-alloy head and a titanium socket without cement. The remaining twenty patients did not have an implant and served as controls. The results of our study showed that, thirty-six months postoperatively, patients who have a well functioning prosthesis with components containing titanium have as much as a threefold increase in the concentration of titanium in the serum and those who have a well functioning prosthesis with cobalt-alloy components have as much as a fivefold and an eightfold increase in the concentrations of chromium in the serum and urine, respectively. The predominant source of the disseminated chromium-degradation products is probably the modular head-neck junction and may be a function of the geometry of the coupling. Passive dissolution of extensively porous-coated cobalt-alloy stems was not found to be a dominant mode of metal release. CLINICAL RELEVANCE: Increased concentrations of circulating metal-degradation products derived from orthopaedic implants may have deleterious biological effects over the long term that warrant investigation. This is a particularly timely concern because of recent clinical trends, including the reintroduction of metal-on-metal bearing surfaces and the increasing popularity of extensively porous-coated devices with large surface areas of exposed metal. Accurate monitoring of the concentrations of metal in the serum and urine after total hip replacement also can provide insights into the mechanisms of metal release. Our findings suggest that fretting corrosion at the head-neck coupling is an important source of metal release that can lead to increased concentrations of chromium in the serum. Determinations of the concentrations of metal in the serum and urine may be useful in the diagnosis of patients who are symptomatic after a total joint replacement as increased levels are indicative of at least one mode of mechanical dysfunction (for example, fretting corrosion) of the device.

Adult↗

Human monocyte/macrophage response to cobalt-chromium corrosion products and titanium particles in patients with total joint replacements.

The responses of human peripheral blood monocytes of 10 normal volunteers and 14 patients with total hip replacements to particles of commercially pure titanium and chromium orthophosphate (a corrosion product from cobalt-chromium alloy implants) were studied. In addition, these phagocytosable particles were added to cultured mononuclear cells isolated from the interfacial membrane of 14 patients with failed implants. Peripheral blood monocytes from patients who had had a total hip replacement produced significantly higher levels of interleukin-1 (both interleukin-1 alpha and interleukin-1 beta) and prostaglandin E2 following particulate stimulation than those from normal volunteers. Supernatants from both titanium and chromium orthophosphate-stimulated peripheral blood monocytes from the volunteers and patients with total hip replacement induced bone resorption (assayed in organ cultures of newborn mouse calvariae) and the proliferation of human fibroblasts. The levels of bone resorption were significantly higher (p < 0.05) in patients with implants than in normal volunteers. There were no significant differences in the responses of cells between patients with focal osteolysis and those without osteolysis. Interfacial membrane mononuclear cells also produced high levels of interleukin-1 alpha, interleukin-1 beta, and prostaglandin E2 and expressed bone resorptive activities following stimulation with either titanium or chromium orthophosphate. More importantly, interfacial membrane mononuclear cells "spontaneously" produced high levels of prostaglandin E2 that were comparable with the response of peripheral blood monocytes stimulated with particulate wear debris. The clinical relevance of this study is 2-fold. First, mononuclear cells from patients with total hip replacement were some-how "sensitized" to metal particles in comparison with mononuclear cells from individuals without an implant. Second, the chromium orthophosphate corrosion product was a potent macrophage/monocyte activator and may contribute to macrophage-mediated osteolysis and aseptic loosening.

Adult↗

Suppression of osteoblast function by titanium particles.

In order to understand the effect of particulate debris on osteoblast function, we studied the effect of different particles, including titanium and polystyrene, on bone collagen mRNA (messenger RNA) with the use of Northern blot hybridization analysis, and we studied the effect of the particles on the biosynthesis of bone collagen with analysis of 3H-proline incorporation and with the Western blot technique. The steady-state levels of mRNA for procollagens alpha1(I) and alpha1(III) were markedly suppressed in human MG-63 osteoblast-like cells exposed to phagocytosable titanium particles that were smaller than three micrometers. Both titanium and polystyrene particles smaller than three micrometers suppressed the expression of the gene that codes for collagen, and the suppression of the expression of the gene was related to the size but not to the composition of the particles. The biosynthesis of both type-I and type-III collagen also was decreased in cells that had been treated with titanium particles. Neither the viability nor the proliferation of cells was affected by particulate debris. These data indicate that phagocytosable titanium particles can significantly suppress the expression of the gene that codes for collagen in osteoblast-like cells (p < 0.05).

Blotting, Northern↗

Primary total hip arthroplasty with a porous-coated acetabular component. Seven-to-ten-year results.

One hundred and eighty-four patients who had been managed, between 1984 and 1985, with a total of 204 consecutive primary total hip arthroplasties with insertion of a Harris-Galante type-I acetabular component without cement were prospectively studied. There were eighty-two men (45 per cent) and 102 women (55 per cent). The mean age at the time of the operation was fifty-two years (range, twenty to eighty-four years). One hundred and fifty-seven patients (173 hips) were available for clinical review at a mean of 104 months (range, seventy-eight to 126 months). At this time, the mean preoperative Harris hip score of 52 points (range, 12 to 79 points) had improved to a mean of 90 points (range, 44 to 100 points). Two patients (two hips) had an exchange of an excessively worn polyethylene liner. One patient (one hip) had debridement and grafting of an area of massive retroacetabular osteolysis. Two stable acetabular components (1 per cent) had been revised at the time of femoral revision. None of the cups had been revised because of aseptic loosening. Radiographic examination of 150 patients (165 hips) at a mean of 104 months (range, seventy-eight to 126 months) revealed that 156 cups (95 per cent) were stable. Eight cups (5 per cent) were considered to be possibly unstable, with a radiolucent line between the prosthesis and the bone that was one millimeter wide or less in at least four of five zones; two of the eight had a complete radiolucent line in all zones. One component, which had been implanted with a bulk allograft from a femoral head, migrated more than two millimeters in the first two years and then stabilized without complication. Osteolysis was seen in seven acetabula (4 per cent) and was limited to the periphery of the cup in six. Survivorship analysis at ten years revealed that the acetabular component had a 99 per cent chance of survival (95 per cent confidence interval, 0.98 to 1.0) with revision or aseptic loosening as the end point and a 97 per cent chance of survival (95 per cent confidence interval, 0.95 to 1.0) with revision, aseptic loosening, or reoperation because of a problem related to the acetabular component as the end point.

Adult↗

The fate of well fixed cemented femoral components left in place at the time of revision of the acetabular component.

The results were reviewed for thirty-seven hips (thirty-five patients) in which a well fixed femoral component that originally had been implanted with use of a so-called first-generation cementing technique was subsequently left in place at the time of revision of the acetabular component without cement. The purpose of the study was to determine the rate of survival of the femoral component and the complications associated with revision of one side of the joint. The femoral components had been in situ for a mean duration of 102 months (range, twelve to 216 months) at the time of the revision of the acetabular component. At the time of follow-up after the revision of the acetabular component, one patient (one femoral component; 3 per cent) had died, twenty-seven femoral components (73 per cent) were in place and radiographically stable, one femoral component had been removed because of infection, six (16 per cent) had been revised because of aseptic loosening, and two (5 per cent) were definitely loose according to radiographic criteria. Excluding the failures and death, the mean duration of follow-up was sixty-five months (range, forty-eight to 121 months). The predicted survival of the femoral component after the revision of the acetabular component was 88 per cent (95 per cent confidence interval, 82 to 94 per cent) at forty-eight months and 78 per cent (95 per cent confidence interval, 67 to 89 per cent) at eighty-eight months. Dislocation occurred in three hips (8 per cent), and a trochanteric non-union occurred in five (14 per cent). The mean rate of linear polyethylene wear did not differ significantly (0.13 as compared with 0.12 millimeter per year; p = 0.74) from that for a comparable group of forty-nine hips that had had a revision of the femoral component without cement and had been followed for a similar duration. The data support the decision to retain a well fixed femoral component that has been implanted with a so-called first-generation cementing technique when the acetabular component subsequently needs a revision. The survival of the femoral component and the risk of associated complications do not appear to be appreciably altered by revision of the other side of the joint.

Acetabulum↗