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1.
Osteoarthr Imaging ; 4(1)2024 Mar.
Article in English | MEDLINE | ID: mdl-38549836

ABSTRACT

Objective: Chronic inflammation and altered walking biomechanics are common after ACL reconstruction (ACLR) and contribute to the development of osteoarthritis. Clinically accessible techniques are needed to monitor inflammation (ultrasound-assessed effusion-synovitis) and walking biomechanics (force-measuring insoles), and they must improve the translation of these assessments and determine whether inflammation and walking biomechanics are related in patients after ACLR. This study aimed to determine the association between ultrasound-detected knee effusion-synovitis and limb loading asymmetries during walking in patients 1-5 years post-ACLR. Design: 15 participants (9 women; age: 26 ± 6yrs; mass: 71 ± 15 kg; height: 173 ± 9 cm; months post-ACLR: 29 ± 13) were included in this cross-sectional study. Knee effusion-synovitis was assessed using a standardized protocol and graded using a validated scoring atlas (0 = absent, 1 = mild, 2 = moderate, 3 = severe) in the ACLR limb. Force-measuring insoles were used to capture the vertical ground reaction force (vGRF) during a one-minute treadmill walking trial. Limb symmetry indices (LSIs) were used to quantify limb loading asymmetry for the peak vGRF and the instantaneous loading rate (vGRF-LR). Spearman correlations determined whether effusion-synovitis grade was associated with peak vGRF and vGRF-LR LSI. Results: Effusion-synovitis was present in the ACLR limb of 13/15 (87 %) participants (Grade 0: n = 2; Grade 1: n = 8; Grade 2: n = 4, Grade 3: n = 1). Effusion-synovitis grade was not significantly associated with peak vGRF LSI (mean±sd: 98.0 ± 5.6; ρ = 0.38, p = 0.162), but was significantly associated with vGRF-LR LSI (98.2 ± 11.4; ρ = 0.55, p = 0.035). Conclusion: Most participants 1-5 years post-ACLR have ultrasound-detected effusion-synovitis. Participants with more severe effusion-synovitis load their ACLR limb more rapidly. This study highlights the utility of clinically accessible techniques in assessing inflammation and walking biomechanics in ACLR patients.

2.
J Orthop Res ; 42(4): 729-736, 2024 Apr.
Article in English | MEDLINE | ID: mdl-37874323

ABSTRACT

This study aimed to create a conversion equation that accurately predicts cartilage magnetic resonance imaging (MRI) T2 relaxation times using ultrasound echo-intensity and common participant demographics. We recruited 15 participants with a primary anterior cruciate ligament reconstruction between the ages of 18 and 35 years at 1-5 years after surgery. A single investigator completed a transverse suprapatellar scan with the ACLR limb in max knee flexion to image the femoral trochlea cartilage. A single reader manually segmented the femoral cartilage cross-sectional area to assess the echo-intensity (i.e., mean gray-scale pixel value). At a separate visit, a T2 mapping sequence with the MRI beam set to an oblique angle was used to image the femoral trochlea cartilage. A single reader manually segmented the cartilage cross-sectional area on a single MRI slice to assess the T2 relaxation time. A stepwise, multiple linear regression was used to predict T2 relaxation time from cartilage echo-intensity and common demographic variables. We created a conversion equation using the regression betas and then used an ICC and Bland-Altman plot to assess agreement between the estimated and true T2 relaxation time. Cartilage ultrasound echo-intensity and age significantly predicted T2 relaxation time (F = 7.33, p = 0.008, R2 = 0.55). When using the new conversion equation to estimate T2 relaxation time from cartilage echo-intensity and age, there was strong agreement between the estimated and true T2 relaxation time (ICC2,k = 0.84). This study provides promising preliminary data that cartilage echo-intensity combined with age can be used as a clinically accessible tool for evaluating cartilage composition.


Subject(s)
Anterior Cruciate Ligament Injuries , Anterior Cruciate Ligament Reconstruction , Cartilage, Articular , Humans , Adolescent , Young Adult , Adult , Knee Joint/pathology , Cartilage, Articular/pathology , Femur/diagnostic imaging , Femur/surgery , Anterior Cruciate Ligament Injuries/surgery , Anterior Cruciate Ligament Reconstruction/methods , Magnetic Resonance Imaging/methods
3.
Ultrasound Med Biol ; 49(12): 2590-2595, 2023 12.
Article in English | MEDLINE | ID: mdl-37770296

ABSTRACT

OBJECTIVE: The aim of the work described here was to evaluate quadriceps muscle ultrasound metrics and common demographic variables to create a conversion equation that validly predicts magnetic resonance imaging (MRI) percent intramuscular fat after anterior cruciate ligament reconstruction (ACLR). METHODS: We recruited 15 participants between the ages of 18 and 35 y who were 1-5 y post-ACLR. For the MRI assessment, we used an iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL) sequence to assess the mid-thigh. A single reader manually segmented the rectus femoris on two consecutive MRI slices using ITK-Snap to estimate the percent intramuscular fat. For the ultrasound assessment, a single investigator captured transverse panoramic ultrasound images of the mid-thigh with the participant positioned supine and the knee flexed to 30°. A separate single reader used ImageJ to manually segment the rectus femoris ultrasound images. Ultrasound metrics included muscle cross-sectional area, echo intensity and subcutaneous fat thickness. A stepwise linear multiple regression was used to develop an equation to predict MRI percent intramuscular fat using the ultrasound metrics and common demographics (i.e., age, sex, height, mass). Additionally, intraclass correlation coefficients (ICC2,k) and Bland-Altman plots were used to assess the agreement between true and estimated percent intramuscular fat. RESULTS: Echo intensity and age significantly predicted MRI intramuscular fat percent (p = 0.003, r2 = 0.62). When using the conversion equation, there was high agreement (ICC2,k = 0.87, 95% confidence interval: 0.62-0.96) between the estimated and true percent intramuscular fat. CONCLUSION: Our patient population-specific conversion equation that uses quadriceps muscle ultrasound echo intensity and age is a valid estimate of MRI percent intramuscular fat.


Subject(s)
Anterior Cruciate Ligament Reconstruction , Quadriceps Muscle , Humans , Adolescent , Young Adult , Adult , Quadriceps Muscle/diagnostic imaging , Quadriceps Muscle/physiology , Knee Joint , Knee , Thigh
4.
Cartilage ; 13(2): 19476035221093069, 2022.
Article in English | MEDLINE | ID: mdl-35438030

ABSTRACT

OBJECTIVE: To validate a semi-automated technique to segment ultrasound-assessed femoral cartilage without compromising segmentation accuracy to a traditional manual segmentation technique in participants with an anterior cruciate ligament injury (ACL). DESIGN: We recruited 27 participants with a primary unilateral ACL injury at a pre-operative clinic visit. One investigator performed a transverse suprapatellar ultrasound scan with the participant's ACL injured knee in maximum flexion. Three femoral cartilage ultrasound images were recorded. A single expert reader manually segmented the femoral cartilage cross-sectional area in each image. In addition, we created a semi-automatic program to segment the cartilage using a random walker-based method. We quantified the average cartilage thickness and echo-intensity for the manual and semi-automated segmentations. Intraclass correlation coefficients (ICC2,k) and Bland-Altman plots were used to validate the semi-automated technique to the manual segmentation for assessing average cartilage thickness and echo-intensity. A dice correlation coefficient was used to quantify the overlap between the segmentations created with the semi-automated and manual techniques. RESULTS: For average cartilage thickness, there was excellent reliability (ICC2,k = 0.99) and a small mean difference (+0.8%) between the manual and semi-automated segmentations. For average echo-intensity, there was excellent reliability (ICC2,k = 0.97) and a small mean difference (-2.5%) between the manual and semi-automated segmentations. The average dice correlation coefficient between the manual segmentation and semi-automated segmentation was 0.90, indicating high overlap between techniques. CONCLUSIONS: Our novel semi-automated segmentation technique is a valid method that requires less technical expertise and time than manual segmentation in patients after ACL injury.


Subject(s)
Anterior Cruciate Ligament Injuries , Cartilage, Articular , Anterior Cruciate Ligament Injuries/diagnostic imaging , Cartilage, Articular/diagnostic imaging , Cartilage, Articular/injuries , Humans , Knee Joint/diagnostic imaging , Reproducibility of Results , Ultrasonography
5.
Clin J Sport Med ; 32(3): e316-e318, 2022 05 01.
Article in English | MEDLINE | ID: mdl-35316824

ABSTRACT

ABSTRACT: Tarsal tunnel syndrome (TTS) typically occurs from extrinsic or intrinsic sources of compression on the tibial nerve. We present 3 cases of patients, all of whom have a prolonged time to diagnosis after evaluation with multiple specialties, with foot pain ultimately secondary to an accessory flexor digitorum longus muscle causing TTS. The literature describing the association between TTS and accessory musculature has been limited to single case reports and frequently demonstrate abnormal electrodiagnostic testing. In our series, 2 cases had normal electrodiagnostic findings despite magnetic resonance imaging (MRI) that later revealed TTS and improvement with eventual resection. A normal electromyogram should not preclude the diagnosis of TTS and MRI of the ankle; it should be considered a useful diagnostic tool when examining atypical foot pain.


Subject(s)
Tarsal Tunnel Syndrome , Ankle , Foot/diagnostic imaging , Humans , Muscle, Skeletal/diagnostic imaging , Pain , Tarsal Tunnel Syndrome/diagnosis , Tarsal Tunnel Syndrome/etiology , Tarsal Tunnel Syndrome/surgery
6.
Skeletal Radiol ; 51(7): 1433-1438, 2022 Jul.
Article in English | MEDLINE | ID: mdl-34988628

ABSTRACT

OBJECTIVE: Glenoid bone loss is estimated using a best-fit circle method and requires software tools that may not be available. Our hypothesis is that a vertical reference line drawn parallel to the long axis of the glenoid and passing through the inflection point of the coracoid and glenoid will represent a demarcation line of approximately 20% of the glenoid. Our aim is to establish a more efficient method to estimate a surgical threshold for glenoid insufficiency. METHODS: Fifty patients with normal glenoid anatomy were randomly chosen from an orthopedic surgeon's database. Two orthopedic surgeons utilized T1-weighted sagittal MRIs and the coracoglenoid line technique to determine the percentage of bony glenoid anterior to vertical line. Two musculoskeletal radiologists measured the same 50 glenoids using the circle technique. Differences were determined using dependent t test. Reliability was compared using interclass correlation coefficient and Kappa. Validity was compared using Pearson correlation coefficient. RESULTS: Mean surface area of the glenoid anterior to the vertical line was on average 21.69% ± 3.12%. Surface area of the glenoid using the circle method was on average 20.86% ± 2.29%. Inter-rater reliability of the circle method was 0.553 (fair). Inter-rater reliability of the vertical line technique was 0.83 (excellent). There was a linear relationship between circle and vertical line measurements, r = 0.704 (moderate to high). CONCLUSION: The coracoglenoid line appears to represent a line of demarcation of approximately 21% of glenoid bone anterior to the coracoglenoid line. Our technique was found to be reliable, valid, and accurate.


Subject(s)
Bone Diseases, Metabolic , Glenoid Cavity , Joint Instability , Shoulder Joint , Glenoid Cavity/diagnostic imaging , Glenoid Cavity/pathology , Humans , Joint Instability/pathology , Magnetic Resonance Imaging , Reproducibility of Results , Scapula/diagnostic imaging , Scapula/pathology , Shoulder Joint/diagnostic imaging , Shoulder Joint/pathology , Tomography, X-Ray Computed/methods
7.
J Orthop Res ; 40(10): 2240-2247, 2022 10.
Article in English | MEDLINE | ID: mdl-35001419

ABSTRACT

Women with anterior cruciate ligament reconstruction report worse pain and knee-related symptoms, and also exhibit biomechanical changes that may be related to knee osteoarthritis (OA) development. This is particularly concerning as symptom state has been previously associated with knee OA development. The purpose of this study was to compare lower extremity walking biomechanics between women (age: 21.40 ± 8.54 years) experiencing clinically significant knee-related symptoms and women with acceptable symptoms 6 months following surgery. Twenty-eight women with history of primary, unilateral anterior cruciate ligament reconstruction who completed a lower extremity walking biomechanics assessment 6 months following surgery were included in this analysis. Women were dichotomized as experiencing acceptable or clinically significant knee symptoms according to Knee injury and OA Outcomes Score cut-offs described by Englund et al. Walking biomechanics were compared between women with clinically significant and acceptable symptoms using one-way analysis of covariances for involved limb biomechanics. Biomechanical variables of interest were: peak vertical ground reaction forces (vGRFs), vGRF loading rates, knee flexion angles, knee extension moments, knee adduction angles, and knee adduction moments, and gait speed. Nearly 60% of women reported clinically significant knee symptoms 6 months postoperative. There were no statistically significant differences between symptom groups for walking biomechanics and gait speed outcomes. These findings suggest patient reported knee symptoms may not be a primary influence on walking biomechanics 6 months following anterior cruciate ligament reconstruction. Though, longitudinal assessment of changes in symptom state and walking biomechanics may be warranted as poorer walking biomechanics and symptoms are indicators of knee OA.


Subject(s)
Anterior Cruciate Ligament Injuries , Anterior Cruciate Ligament Reconstruction , Osteoarthritis, Knee , Adolescent , Adult , Anterior Cruciate Ligament Injuries/surgery , Biomechanical Phenomena , Child , Female , Gait , Humans , Knee Joint/surgery , Osteoarthritis, Knee/surgery , Walking , Young Adult
8.
ACS Biomater Sci Eng ; 7(2): 718-726, 2021 02 08.
Article in English | MEDLINE | ID: mdl-33449622

ABSTRACT

Clinical effectiveness of implantable medical devices would be improved with in situ monitoring to ensure device positioning, determine subsequent damage, measure biodegradation, and follow healing. While standard clinical imaging protocols are appropriate for diagnosing disease and injury, these protocols have not been vetted for imaging devices. This study investigated how radiologists use clinical imaging to detect the location and integrity of implanted devices and whether embedding nanoparticle contrast agents into devices can improve assessment. To mimic the variety of devices available, phantoms from hydrophobic polymer films and hydrophilic gels were constructed, with and without computed tomography (CT)-visible TaOx and magnetic resonance imaging (MRI)-visible Fe3O4 nanoparticles. Some phantoms were purposely damaged by nick or transection. Phantoms were implanted in vitro into tissue and imaged with clinical CT, MRI, and ultrasound. In a blinded study, radiologists independently evaluated whether phantoms were present, assessed the type, and diagnosed whether phantoms were damaged or intact. Radiologists identified the location of phantoms 80% of the time. However, without incorporated nanoparticles, radiologists correctly assessed damage in only 54% of cases. With an incorporated imaging agent, the percentage jumped to 86%. The imaging technique which was most useful to radiologists varied with the properties of phantoms. With benefits and drawbacks to all three imaging modalities, future implanted devices should be engineered for visibility in the modality which best fits the treated tissue, the implanted device's physical location, and the type of required information. Imaging protocols should also be tailored to best exploit the properties of the imaging agents.


Subject(s)
Magnetic Resonance Imaging , Tomography, X-Ray Computed , Phantoms, Imaging , Prostheses and Implants , Ultrasonography
9.
Arthroscopy ; 32(1): 63-8, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26343942

ABSTRACT

PURPOSE: To determine differences in tibial tubercle-trochlear groove (TT-TG) distance between patients with a history of noncontact anterior cruciate ligament (ACL) injury and an uninjured control group. METHODS: MRI studies of 60 patients (age range, 14 to 25 years) with ACL-deficient (ACLD) knees were compared with 60 intact-ACL controls. All patients underwent MRI after a noncontact sports injury. TT-TG distances were measured on proton density-weighted axial images. Independent t-tests were used to determine differences in TT-TG distance between the ACLD and control groups. RESULTS: The mean TT-TG distance in the ACLD group was 12.07 mm (95% confidence interval [CI], 11.11 to 13.02), compared with 10.44 mm (95% CI, 9.64 to 11.24) in the control group. The mean TT-TG distance in the male ACLD group was 12.95 mm (95% CI, 11.39 to 14.51), compared with 10.87 mm (95% CI, 9.52 to 12.21) in the male control group. The mean TT-TG distance in the female ACLD group was 11.48 mm (95% CI, 10.24 to 12.71), compared with 10.04 mm (95% CI, 9.06 to 11.02) in the female control group. There were statistically significant differences in TT-TG distance between the ACLD and control groups (P = .011) and between the male ACLD and control groups (P = .041). CONCLUSIONS: In adolescents and young adults, the TT-TG distance was statistically larger in knees with noncontact ACL tears than in intact-ACL control knees. When the groups were stratified on the basis of sex, only the male patients showed a statistical difference, with a 2.08 mm increase in TT-TG distance between the ACLD and intact-ACL patients. No difference in TT-TG distance was found between the ACLD and control groups for female patients. Despite the findings of this study, the clinical significance of an increased TT-TG distance as an isolated risk factor for noncontact ACL injury remains unanswered. LEVEL OF EVIDENCE: Level III, case-control study.


Subject(s)
Anterior Cruciate Ligament Injuries , Patellofemoral Joint/anatomy & histology , Tibia/anatomy & histology , Adolescent , Adult , Athletic Injuries , Case-Control Studies , Female , Humans , Magnetic Resonance Imaging , Male , Retrospective Studies , Sex Factors , Young Adult
10.
J Orthop Res ; 33(12): 1835-45, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26147652

ABSTRACT

Subchondral bone is thought to play a significant role in the initiation and progression of the post-traumatic osteoarthritis. The goal of this study was to document changes in tibial and femoral subchondral bone that occur as a result of two lapine models of anterior cruciate ligament injury, a modified ACL transection model and a closed-joint traumatic compressive impact model. Twelve weeks post-injury bones were scanned via micro-computed tomography. The subchondral bone of injured limbs from both models showed decreases in bone volume and bone mineral density. Surgical transection animals showed significant bone changes primarily in the medial hemijoint of femurs and tibias, while significant changes were noted in both the medial and lateral hemijoints of both bones for traumatic impact animals. It is believed that subchondral bone changes in the medial hemijoint were likely caused by compromised soft tissue structures seen in both models. Subchondral bone changes in the lateral hemijoint of traumatic impact animals are thought to be due to transmission of the compressive impact force through the joint. The joint-wide bone changes shown in the traumatic impact model were similar to clinical findings from studies investigating the progression of osteoarthritis in humans.


Subject(s)
Bone and Bones/diagnostic imaging , Osteoarthritis/diagnostic imaging , Animals , Anterior Cruciate Ligament/surgery , Bone Density , Cartilage, Articular/diagnostic imaging , Disease Models, Animal , Disease Progression , Female , Femur/diagnostic imaging , Femur/pathology , Hindlimb/pathology , Magnetic Resonance Imaging , Male , Menisci, Tibial/surgery , Rabbits , Tibia/diagnostic imaging , Tibia/pathology , X-Ray Microtomography
11.
J Biomech ; 48(2): 246-53, 2015 Jan 21.
Article in English | MEDLINE | ID: mdl-25523754

ABSTRACT

The objective of this study was to induce anterior cruciate ligament (ACL) and meniscal damage, via a single tibiofemoral compressive impact, in order to document articular cartilage and meniscal changes post-impact. Tibiofemoral joints of Flemish Giant rabbits were subjected to a single blunt impact that ruptured the ACL and produced acute meniscal damage. Animals were allowed unrestricted cage activity for 12 weeks before euthanasia. India ink analysis of the articular cartilage revealed higher degrees of surface damage on the impacted tibias (p=0.018) and femurs (p<0.0001) compared to controls. Chronic meniscal damage was most prevalent in the medial central and medial posterior regions. Mechanical tests revealed an overall 19.4% increase in tibial plateau cartilage thickness (p=0.026), 34.8% increase in tibial plateau permeability (p=0.054), 40.8% increase in femoral condyle permeability (p=0.029), and 20.1% decrease in femoral condyle matrix modulus (p=0.012) in impacted joints compared to controls. Both instantaneous and equilibrium moduli of the lateral and medial menisci were decreased compared to control (p<0.02). Histological analyses revealed significantly increased presence of fissures in the medial femur (p=0.036). In both meniscus and cartilage there was a significant decrease in GAG coverage for the impacted limbs. Based on these results it is clear that an unattended combined meniscal and ACL injury results in significant changes to the soft tissues in this experimental joint 12 weeks post-injury. Such changes are consistent with a clinical description of mid to late stage PTOA of the knee.


Subject(s)
Cartilage, Articular/pathology , Knee Injuries/pathology , Mechanical Phenomena , Menisci, Tibial/pathology , Animals , Anterior Cruciate Ligament/pathology , Anterior Cruciate Ligament Injuries , Biomechanical Phenomena , Cartilage, Articular/injuries , Compressive Strength , Femur/injuries , Femur/pathology , Rabbits , Rupture/pathology , Tibia/injuries , Tibia/pathology , Tibial Meniscus Injuries , Time Factors
12.
Am J Sports Med ; 40(6): 1337-41, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22422932

ABSTRACT

BACKGROUND: Acetabular labral tears as a source of potential hip pain have received a great deal of attention in recent literature. The gold standard for identifying acetabular labral tears is hip arthroscopy, but recent advances in optimized, noncontrast magnetic resonance imaging (MRI) have proven effectiveness in identifying intra-articular hip pathological changes without the invasive nature of hip arthroscopy or gadolinium-enhanced arthrography. There are little data in the literature on imaging results in an asymptomatic population. PURPOSE: The objective of this study was to use an optimized, noncontrast 1.5-T MRI protocol to identify hip abnormalities, including paralabral cysts, in asymptomatic volunteers. STUDY DESIGN: Case series (prevalence); Level of evidence, 4. METHODS: In this study, 42 hips in asymptomatic patients with an average age of 34 years (range, 27-43 years) were prospectively imaged with optimized, noncontrast 1.5-T MRI scans. Two fellowship-trained musculoskeletal radiologists interpreted the scans at 2 different points in time and commented on the presence of labral abnormalities including paralabral cysts. The results were analyzed for both interobserver and intraobserver reliability. RESULTS: Acetabular paralabral cysts were identified in 11 of 42 (26.2%) and 9 of 42 (21.4%) hips by the 2 respective radiologists, with an interobserver reliability of 90.5% (κ = .74) and intraobserver reliability of 95.2% (κ = .87). In addition, acetabular labral tears were identified in 36 of 42 (85.7%) and 34 of 42 (80.9%) hips, with an interobserver reliability of 90.5% (κ = .70) and intraobserver reliability of 95.2% (κ = .83). CONCLUSION: Utilizing an optimized, noncontrast 1.5-T MRI protocol, we report the previously undescribed prevalence of acetabular labral pathological abnormalities and paralabral cysts in a young, asymptomatic population. This emphasizes the importance of correlating patient symptoms with history and physical examination when evaluating patients with hip pain and radiographic abnormalities as defined by MRI criteria. These data demonstrate that labral tears can occur without symptoms.


Subject(s)
Acetabulum/pathology , Cysts/pathology , Hip Injuries/pathology , Hip/pathology , Magnetic Resonance Imaging/methods , Acetabulum/injuries , Adolescent , Adult , Cysts/diagnosis , Female , Hip/abnormalities , Hip Injuries/diagnosis , Hip Injuries/epidemiology , Humans , Male , Middle Aged , Prevalence , Reproducibility of Results , Young Adult
13.
Semin Musculoskelet Radiol ; 12(1): 62-71, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18382945

ABSTRACT

Stress fractures are common, representing the final stage in a continuum of bone response to continued mechanical damage. Encompassing fatigue- and insufficiency-type fractures, stress fractures of the pelvis are likely underreported. Radiographs are insensitive to stress injuries, particularly those in the pelvis, whereas scintigraphy and magnetic resonance imaging are exquisitely sensitive. In this article we discuss the pathophysiology and imaging appearances of stress injuries of the pelvis and sacrum. Relevant literature regarding risk factors, problem-solving issues, and an imaging algorithm are discussed, with the goal of improving accuracy in the diagnosis of these common injuries.


Subject(s)
Athletic Injuries/diagnosis , Diagnostic Imaging , Fractures, Stress/diagnosis , Pelvic Bones/injuries , Pelvic Pain/diagnosis , Athletic Injuries/physiopathology , Fractures, Stress/physiopathology , Humans , Pelvic Pain/physiopathology
15.
Infect Immun ; 72(3): 1693-9, 2004 Mar.
Article in English | MEDLINE | ID: mdl-14977977

ABSTRACT

The pathogenic yeast Cryptococcus neoformans produces a laccase enzyme (CNLAC1), which catalyzes the synthesis of melanin in the presence of phenolic compounds. A number of genes have been implicated in the regulation of laccase and melanization, including IPC1, GPA1, MET3, and STE12. Albino mutants derived from random mutagenesis techniques may contain mutations in genes that regulate multiple virulence factors, including CNLAC1. The goal of our study is to investigate the role of CNLAC1 in virulence and evasion of pulmonary host defenses after infection via the respiratory tract. Using a set of congenic laccase-positive (2E-TUC-4) and laccase-deficient (2E-TU-4) strains, we found that both strains are avirulent at a lower dose (10(4) CFU/mouse) in mice. After the infectious dose was increased to 10(6) CFU/mouse, 70% mortality was observed in mice infected with 2E-TUC-4 compared to no mortality in mice infected with 2E-TU-4 at day 30 postinfection. This observation confirms the requirement for CNLAC1 in virulence. Interestingly, we observed no differences between the two strains in pulmonary growth or in elicitation of cellular immune responses in the lung. The only measurable defect of 2E-TU-4 was in dissemination to extrapulmonary sites. To examine the role of CNLAC1 in dissemination, mice were infected intravenously. By week 3 postinfection, equal numbers of strains 2E-TUC-4 and 2E-TU-4 were recovered from the brain and spleen. This observation indicates that CNLAC1 facilitates escape from the lung, but not growth in the lungs or brain, and suggests a novel role for CNLAC1 in virulence during an infection aquired via the respiratory tract.


Subject(s)
Cryptococcosis/etiology , Cryptococcus neoformans/enzymology , Cryptococcus neoformans/pathogenicity , Laccase/physiology , Lung Diseases, Fungal/etiology , Animals , Cryptococcosis/microbiology , Cryptococcosis/pathology , Cryptococcus neoformans/genetics , Female , Genes, Fungal , Immunity, Cellular , Laccase/genetics , Lung/immunology , Lung/microbiology , Lung/pathology , Lung Diseases, Fungal/microbiology , Lung Diseases, Fungal/pathology , Mice , Mice, Inbred CBA , Mutation , Virulence/genetics , Virulence/physiology
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