Neonatal screening for severe primary immunodeficiency diseases using high-throughput triplex real-time PCR

Severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia (XLA) are inborn errors of immune function that require prompt diagnosis and treatment to prevent life-threatening infections. The lack of functional T or B lymphocytes in these diseases serves as a diagnostic criterion and can be applied to neonatal screening. A robust triplex PCR method for quantitation of T-cell receptor excision circles (TRECs) and κ-deleting recombination excision circles (KRECs), using a single Guthrie card punch, was developed and validated in a cohort of 2560 anonymized newborn screening cards and in 49 original stored Guthrie cards from patients diagnosed with SCID, XLA, ataxia-telangiectasia, Nijmegen-breakage-syndrome, common variable immunodeficiency, immunoglobulin A deficiency, or X-linked hyper-IgM syndrome. Simultaneous measurement of TREC and KREC copy numbers in Guthrie card samples readily identified patients with SCID, XLA, ataxia-telangiectasia and Nijmegen-breakage-syndrome and thus facilitates effective newborn screening for severe immunodeficiency syndromes characterized by the absence of T or B cells.     

Assessing the validity of corneal power estimation using conventional keratometry for intraocular lens power calculation in eyes with Fuch’s dystrophy undergoing Descemet membrane endothelial keratoplasty

Graefe's Archive for Clinical and Experimental Ophthalmology - The present retrospective study was designed to test the hypothesis that the postoperative posterior to preoperative anterior...     

Myocardial Uptake and Release of Substrates in Type II Diabetics Undergoing Coronary Surgery

Objective - Knowledge about myocardial metabolism in diabetic patients is limited, and even more so in association with myocardial ischaemia and cardiac surgery. This study investigates myocardial substrate utilization in type II diabetic patients after elective coronary artery bypass graft surgery (CABG). Design - Myocardial substrate utilization in 10 type II diabetic patients was assessed during the first hours after elective CABG with the coronary sinus catheter technique. Results - No significant myocardial uptake of carbohydrates was observed. Free fatty acids were extracted throughout the study period but uptake was not related to arterial levels. As arterial levels of beta-hydroxybutyric acid tended to increase a significant myocardial uptake emerged. The average extraction rate of beta-hydroxybutyric acid was 31% with a peak of 42%. A comparable extraction rate of glutamate was also found whereas alanine was released from the heart. Conclusion - Free fatty acids were the main source of energy for the heart whereas uptake of carbohydrates was restricted. The high extraction rates of beta-hydroxybutyric acid and glutamate suggest an adaptive role for these substrates during this unfavourable metabolic state for the post-ischaemic diabetic heart.     

Catastrophic failure of an uncemented acetabular component due to high wear and osteolysisAn analysis of 154 Omnifit prostheses with mean 6-year follow-up

Background?The purposes of this study were (1) to evaluate the wear pattern of the hydroxyapatite-coated “Dual Radius” Omnifit cup, (2) to investigate whether wear is correlated to any demographic or prosthesis-related factors, and (3) to describe micromotion of both the cup and the stem.Patients and methods?154 hips were implanted between 1990 and 1996 and followed for an average of 6 years. Wear was measured according to the “Charnley-duo” method and, in 79 hips, with radiostereometry (RSA). RSA was also used to evaluate micromotion. We analyzed the femoral heads using scanning electron microscopy, energy dispersive X-ray spectroscopy and an atomic force microscope.Result?66 cups were revised and had a mean annual wear of 0.32?mm compared to 0.12?mm in hips not revised. Osteolytic processes were observed in 35 hips but at revision osteolysis was present in 51 cases. 43/66 sockets were loose. Micromotion evaluated by RSA, weight, age, side, size of cup, screws, polyethylene thickness or shelf-life of the polyethylene did not correlate to wear, whereas male gender did.Interpretation?It is still unclear why about half of our cases had an abnormal wear rate. Annual wear exceeding 0.2?mm is prognostic of late failure and should be considered a warning sign.     

A PROSPECTIVE RANDOMISED STUDY (NAMED SVEA) OF THREE METHODS OF DELAYED BREAST RECONSTRUCTION. STUDY DESIGN,PATIENTS' PREOPERATIVE PROBLEMS AND EXPECTATIONS

A randomised study named SVEA to compare three methods for delayed breast reconstruction was initiated in 1994. The methods were: the lateral thoracodorsal flap, the latissimus dorsi flap, and the pedicled transverse rectus abdominis muscle flap. In the present paper we describe the study design, primary endpoints, and inclusion and exclusion criteria. Results from preoperative questionnaires about problems after mastectomy and expectations of the reconstruction are presented. The preoperative questionnaires, completed before randomisation, included a health related quality of life questionnaire, the SF-36, and a questionnaire concerning the impact of breast loss and expectations of the reconstruction. A total of 87 patients have been randomised in SVEA and 30 patients have been followed up outside the randomised study, comprising a reference group. The results from preoperative questionnaires, completed by 106 of the total 117 women, showed that they encountered many problems after mastectomy, primarily about feeling mutilated and being bothered socially. They held high expectations on the breast reconstruction in these areas. The women in the present study did not differ from Swedish women in general regarding health related quality of life, with two exceptions: the study sample scored lower on physical functioning and mental health.     

QT correction using Bazett’s formula remains preferable in long QT syndrome type 1 and 2

BackgroundThe heart rate (HR) corrected QT interval (QTc) is crucial for diagnosis and risk stratification in the long QT syndrome (LQTS). Although its use has been questioned in some contexts, Bazett''s formula has been applied in most diagnostic and prognostic studies in LQTS patients. However, studies on which formula eliminates the inverse relation between QT and HR are lacking in LQTS patients.We therefore determined which QT correction formula is most appropriate in LQTS patients including the effect of beta blocker therapy and an evaluation of the agreement of the formulae when applying specific QTc limits for diagnostic and prognostic purposes.MethodsAutomated measurements from routine 12‐lead ECGs from 200 genetically confirmed LQTS patients from two Swedish regions were included (167 LQT1, 33 LQT2). QT correction was performed using the Bazett, Framingham, Fridericia, and Hodges formulae. Linear regression was used to compare the formulae in all patients, and before and after the initiation of beta blocking therapy in a subgroup (n = 44). Concordance analysis was performed for QTc ≥ 480 ms (diagnosis) and ≥500 ms (prognosis).ResultsThe median age was 32 years (range 0.1–78), 123 (62%) were female and 52 (26%) were children ≤16 years. Bazett''s formula was the only method resulting in a QTc without relation with HR. Initiation of beta blocking therapy did not alter the result. Concordance analyses showed clinically significant differences (Cohen''s kappa 0.629–0.469) for diagnosis and prognosis in individual patients.ConclusionBazett''s formula remains preferable for diagnosis and prognosis in LQT1 and 2 patients.     

Validation of plasma thyroxine and triiodothyronine methods on the ADVIA Centaur® XP

Standardization programs for thyroid hormones have revealed bias between immunochemical methods and the reference method ED-ID-LC/MS. Lack of standardization between methods, suboptimal reference intervals and replacement of serum with plasma may compromise the capability of the immunochemical thyroid methods to diagnose thyroid disease. To accommodate the demand for faster turn-around times for laboratory replies, we replaced serum with plasma on some serum CE marked thyroid methods. This forced us to do on-board analytical correction for the plasma total T4 (TT4) method on ADVIA Centaur^® XP. We, next, validated the capability of the ADVIA Centaur^® XP thyroid methods on plasma by (1) first carrying out a prospective method comparison with the ED-ID-LC/MS reference method using collected plasma samples, (2) we verified the clinical reference intervals by analyzing collected plasma samples from healthy individuals, and (3) retrospectively compared laboratory results from two different time periods using serum TT4 and serum total triiodothyronine (TT3) versus plasma free thyroxine (FT4) and plasma TT3, respectively, to diagnose thyroid disease. The plasma FT4 method displayed a negative concentration-dependent bias against the reference method. This bias was apparently counteracted by a fitted reference interval for the plasma FT4 method. Indeed, overt hyperthyroid disease was found in 1.0% and 1.1% of the cases using serum and plasma and overt hypothyroid condition were in 1.3% and 0.6% of the cases using serum and plasma, respectively. In conclusion, the ADVIA Centaur^® XP FT4 method displayed a negative bias at high plasma FT4 concentrations against the reference method, but the diagnostic performance was not compromised due to a fitted reference interval.