ABSTRACT
BACKGROUND: We aimed to clarify the emerging epigenetic landscape in a group of genes classified as "modifier genes" of the ß-type globin genes (HBB cluster), known to operate in trans to accomplish the two natural developmental switches in globin expression, from embryonic to fetal during the first trimester of conception and from fetal to adult around the time of birth. The epigenetic alterations were determined in adult sickle cell anemia (SCA) homozygotes and SCA/ß-thalassemia compound heterozygotes of Greek origin, who are under hydroxyurea (HU) treatment. Patients were distinguished in HU responders and HU non-responders (those not benefited from the HU) and both, and in vivo and in vitro approaches were implemented. RESULTS: We examined the CpG islands' DNA methylation profile of BCL11A, KLF1, MYB, MAP3K5, SIN3A, ZBTB7A, and GATA2, along with γ-globin and LRF/ZBTB7A expression levels. In vitro treatment of hematopoietic stem cells (HSCs) with HU induced a significant DNA hypomethylation pattern in ZBTB7A (p*, 0.04) and GATA2 (p*, 0.03) CpGs exclusively in the HU non-responders. Also, this group of patients exhibited significantly elevated baseline methylation patterns in ZBTB7A, before the HU treatment, compared to HU responders (p*, 0.019) and to control group of healthy individuals (p*, 0.021), which resembles a potential epigenetic barrier for the γ-globin expression. γ-Globin expression in vitro matched with detected HbF levels during patients' monitoring tests (in vivo) under HU treatment, implying a good reproducibility of the in vitro HU epigenetic effect. LRF/ZBTB7A expression was elevated only in the HU non-responders under the influence of HU. CONCLUSIONS: This is one of the very first pharmacoepigenomic studies indicating that the hypomethylation of ZBTB7A during HU treatment enhances the LRF expression, which by its turn suppresses the HbF resumption in the HU non-responders. Its role as an epigenetic regulator of hemoglobin switching is also supported by the wide distribution of ZBTB7A-binding sites within the 5' CpG sequences of all studied human HBB cluster "modifier genes." Also, the baseline methylation level of selective CpGs in ZBTB7A and GATA2 could be an indicator of the negative HU response among the ß-type hemoglobinopathy patients.
Subject(s)
Anemia, Sickle Cell/genetics , DNA Methylation/genetics , DNA-Binding Proteins/genetics , Hydroxyurea/administration & dosage , Transcription Factors/genetics , beta-Thalassemia/genetics , Anemia, Sickle Cell/blood , Anemia, Sickle Cell/drug therapy , Anemia, Sickle Cell/pathology , Carrier Proteins/genetics , DNA Methylation/drug effects , Female , GATA2 Transcription Factor/genetics , Gene Expression Regulation/drug effects , Heterozygote , Humans , Hydroxyurea/adverse effects , Kruppel-Like Transcription Factors/genetics , MAP Kinase Kinase Kinase 5/genetics , Male , Nuclear Proteins/genetics , Repressor Proteins/genetics , Sin3 Histone Deacetylase and Corepressor Complex , beta-Globins/genetics , beta-Thalassemia/blood , beta-Thalassemia/drug therapy , beta-Thalassemia/pathologyABSTRACT
PURPOSE: Aim of this study was to record and compare the functional and activity level as well as the manifestations of osteoarthritis after isolated ACL ruptures between patients with conservative treatment and ACL reconstruction with hamstrings tendon graft. METHODS: Thirty-two patients diagnosed with ACL rupture were recorded. Clinical examination included the Tegner and Lysholm activity scale, the International Knee Documentation Committee Subjective Form and KT-1000 arthrometer. Narrowing of the medial and lateral joint spaces was assessed using the IKDC knee examination score. RESULTS: Median follow-up was 10.3 years (range 10-11). Fifteen patients were conservatively treated (median age 33 years, range 25-39). Seventeen patients were operated (median age 31 years, range 20-36). There was significant difference between the mean values of IKDC scores in favour of the ACL-reconstruction group of patients, 86.8 (SD 6.5) versus 77.5 (SD 13.8), respectively (p = 0.04). The mean value of anteroposterior tibial translation was 1.5 mm (SD 0.2) for ACL-reconstruction group of patients, while the corresponding mean value for ACL-conservative group was 4.5 mm (SD 0.5), p = 0.03. Four patients in ACL-reconstruction group had radiological findings of grade C or D according to IKDC form. In ACL-conservative group, five patients presented similar signs (n.s.). CONCLUSIONS: ACL reconstruction using hamstrings autograft resulted in better functional outcome and laxity measurements than ACL-conservative management. However, the incidence of radiological osteoarthritis was similar between the two groups and independent on the pre-operative grade of laxity and functional status of the patients. Equally, bone bruises were not found as a risk factor for the development of osteoarthritis after ACL rupture. LEVEL OF EVIDENCE: Prospective randomized study, Level II.
Subject(s)
Anterior Cruciate Ligament Injuries/therapy , Anterior Cruciate Ligament Reconstruction/adverse effects , Osteoarthritis, Knee/etiology , Postoperative Complications/etiology , Adult , Anterior Cruciate Ligament Injuries/surgery , Anterior Cruciate Ligament Reconstruction/methods , Female , Follow-Up Studies , Humans , Joint Instability/etiology , Male , Osteoarthritis, Knee/diagnostic imaging , Prospective Studies , Radiography , Rupture/surgery , Tendons/transplantation , Transplantation, Autologous , Treatment Outcome , Young AdultABSTRACT
Although postoperative septic arthritis is rare after ACL reconstruction, it carries a high morbidity that results in poor clinical outcome. Despite low incidence, it is important to recognize that infection and treat it without delay because of devastating consequences, such as loss of hyaline cartilage and arthrofibrosis, in order to avoid osteoarthritis development and near full range of motion achieved. Herein we discuss the pathogenesis, risk factors, clinical presentation, diagnostic evaluation, treatment protocols and complications of septic knee arthritis after ACL reconstruction.
Subject(s)
Anterior Cruciate Ligament/surgery , Arthritis, Infectious/etiology , Plastic Surgery Procedures , Surgical Wound Infection/etiology , Arthritis, Infectious/diagnosis , Arthritis, Infectious/therapy , Diagnosis, Differential , Humans , Plastic Surgery Procedures/adverse effects , Risk Factors , Surgical Wound Infection/diagnosis , Surgical Wound Infection/therapyABSTRACT
Experimental models for studying transplantation have up to now been unable to isolate reperfusion injury with minimal surgical manipulation and without the interference of graft rejection. Six pigs were subjected to left hilum preparation only (control group), and eight pigs were subjected to left hilum preparation plus in situ cooling ischemia and reperfusion of the lung (experimental group). The hilum was dissected free from other tissues in both groups. Lung preservation was achieved by antegrade flush perfusion via the left pulmonary artery. Pulmonary veins were clamped at the left atrium and a vent was created. The left main bronchus was clamped. Lung temperature was maintained at 4 degrees -8 degrees C, while core temperature was kept at 38 degrees C. After 3 hrs of cold ischemia the clamps were removed and the lung was reperfused. Elevated pulmonary vascular resistance and local and systemic aspects of ischemia-reperfusion syndrome were consistently reproduced. This large-animal model of in situ unilateral lung cold ischemia with warm reperfusion proved to be very reliable in reproducing all aspects of ischemia-reperfusion injury. It excludes the interference of rejection and extensive surgical manipulation. We therefore propose its use in experimental studies investigating pharmaceutical or cooling modifications affecting lung ischemia-reperfusion outcomes.
