Evaluation of Autophagy Process in Differentiation of Human Induced Pluripotent Stem Cells toward Insulin Producing Cells.

Background: Autophagy is an intracellular self-degradative homeostasis process which eliminates undesirable and harmful macromolecules and organelles. Autophagy is also involved in self-renewal and differentiation of induced pluripotent stem cell (iPSCs). Objective: In this study, we investigated the expression profile of autophagy marker genes in human iPSCs during their differentiation induction toward insulin producing ß-like cells. Methods: Human iPSC line, R1-hiPSC1, was used for differentiation induction toward ß-like cells. The mRNA expression of Nanog, OCT4 (pluripotency markers), SOX17, FOXA2 (endodermic markers), PTF1A, NKX6.1 (exocrine/endocrine determinants), and PDX1 were measured during differentiation stages. Autophagy was monitored by genes expression study of four autophagy markers, MAP1LC3B, BECN1, SQSTM1/P62 and ATG5, along with protein expression profile of LC3b-II during differentiation stages. Results: The mRNA expression measurement of pluripotency, endoderm and exocrine/endocrine marker genes confirmed that hiPSCs skipped pluripotency, differentiated into endoderm, passed through the pancreatic lineage commitment stage and successfully generated insulin producing ß-like cells. Expression profile of autophagy genes during differentiation stages indicated the decreased expression levels at the early stages (EB and MEI) and then increased at the definitive endoderm stages (DEI 1, DEI 2 and DE) followed by a subtractive pattern toward the end of differentiation. The results of protein expression of LC3b-II were consistent with gene expression data. Conclusion: This study demonstrated the high contribution of key autophagy genes/proteins during the differentiation of hiPSC toward ß-like cells. The enhanced autophagy levels were a prominent feature of early stages of differentiation and DE rather than the later stages.

Focus on radioiodine-131 biokinetics: the influence of methylprednisolone on intratherapeutic effective half-life of 131I during radioiodine therapy of Graves' disease.

AIM: Radioiodine therapy (RIT) may trigger the development of Graves' ophthalmopathy (GO) or exacerbate pre-existing subclinical GO. Therefore, glucocorticoid administration is recommended for patients with pre-existing GO. Aim of this study was to analyze the influence of glucocorticoid therapy with methylprednisolone on intratherapeutic effective half-life (EHL) of radioiodine-131 in patients with Graves' disease (GD) as recent studies showed an effect for prednisolone. METHODS: In a retrospective study, 264 patients with GD who underwent RIT without any additional antithyroid medication were evaluated. Intrathyroidal EHL was determined pre- and intratherapeutically. Patients with co-existing GO (n = 43) received methylprednisolone according to a fixed scheme starting 1 day prior to RIT, patients without GO (n = 221) did not receive any protective glucocorticoid medication. The ratios of EHL during RIT and during radioiodine uptake test (RIUT) were compared. RESULTS: Patients receiving methylprednisolone showed a slight decrease of the mean EHL from 5.63 d (RIUT) to 5.39 d (RIT) (p > 0.05). A comparable result was obtained in patients without glucocorticoids (5.71 d (RIUT) to 5.47 d (RIT); p > 0.05). The ratios of the EHL between RIT and RIUT failed to show a significant difference between the two groups. EHL is therefore not significantly influenced by an additional protective treatment with methylprednisolone. CONCLUSIONS: In the present study a decreased intrathyroidal EHL under glucocorticoid medication with methylprednisolone could not be detected. Therefore, co-medication with methylprednisolone in patients with GO may be preferred to avoid an intratherapeutic decrease of EHL by accompanying protective glucocorticoides.

Correction for hyperfunctioning radiation-induced stunning (CHRIS) in benign thyroid diseases.

PURPOSE: Radioiodine-131 treatment has been a well-established therapy for benign thyroid diseases for more than 75 years. However, the physiological reasons of the so-called stunning phenomenon, defined as a reduced radioiodine uptake after previous diagnostic radioiodine administration, are still discussed controversially. In a recent study, a significant dependence of thyroid stunning on the pre-therapeutically administered radiation dose could be demonstrated in patients with goiter and multifocal autonomous nodules. A release of thyroid hormones to the blood due to radiation-induced destruction of thyroid follicles leading to a temporarily reduced cell metabolism was postulated as possible reason for this indication-specific stunning effect. Therefore, the aim of this study was to develop dose-dependent correction factors to account for stunning and thereby improve precision of radioiodine treatment in these indications. METHODS: A retrospective analysis of 313 patients (135 with goiter and 178 with multifocal autonomous nodules), who underwent radioiodine uptake testing and radioiodine treatment, was performed. The previously determined indication-specific values for stunning of 8.2% per Gray in patients with multifocal autonomous nodules and 21% per Gray in patients with goiter were used to modify the Marinelli equation by the calculation of correction factors for hyperfunctioning radiation-induced stunning (CHRIS). Subsequently, the calculation of the required activity of radioiodine-131 to obtain an intra-therapeutic target dose of 150 Gy was re-evaluated in all patients. Furthermore, a calculation of the hypothetically received target dose by using the CHRIS-calculated values was performed and compared with the received target doses. RESULTS: After integrating the previously obtained results for stunning, CHRIS-modified Marinelli equations could be developed for goiter and multifocal autonomous nodules. For patients with goiter, the mean value of administered doses calculated with CHRIS was 149 Gy and did not differ from the calculation with the conventional Marinelli equation of 152 Gy with statistical significance (p = 0.60). However, the statistical comparison revealed a highly significant improvement (p < 0.000001) of the fluctuation range of the results received with CHRIS. Similar results were obtained in the subgroup of patients with multifocal autonomous nodules. The mean value of the administered dose calculated with the conventional Marinelli equation was 131 Gy and therefore significantly below the CHRIS-calculated radiation dose of 150 Gy (p < 0.05). Again, the fluctuation range of the CHRIS-calculated radiation dose in the target volume was significantly improved compared with the conventional Marinelli equation (p < 0.000001). CONCLUSIONS: With the presented CHRIS equation it is possible to calculate a required individual stunning-independent radioiodine activity for the first time by only using data from the radioiodine uptake testing. The results of this study deepen our understanding of thyroid stunning in benign thyroid diseases and improve precision of dosimetry in radioiodine-131 therapy of goiter and multifocal autonomous nodules.

Improved Drought Tolerance by AMF Inoculation in Maize (Zea mays) Involves Physiological and Biochemical Implications.

The role of arbuscular mycorrhizal fungus (AMF, Glomus versiforme) in amelioration of drought-induced effects on growth and physio-biochemical attributes in maize (Zea mays L.) was studied. Maize plants were exposed to two drought regimes, i.e., moderate drought (MD) and severe drought (SD), with and without AMF inoculation. Drought at both levels reduced plant height, and chlorophyll and carotenoid content, thereby impeding photosynthesis. In addition, drought stress enhanced the generation of toxic reactive oxygen species (ROS), including H2O2, resulting in membrane damage reflected as increased electrolyte leakage and lipid peroxidation. Such negative effects were much more apparent under SD conditions that those of MD and the control, however, AMF inoculation significantly ameliorated the deleterious effects of drought-induced oxidative damage. Under control conditions, inoculation of AMF increased growth and photosynthesis by significantly improving chlorophyll content, mineral uptake and assimilation. AMF inoculation increased the content of compatible solutes, such as proline, sugars and free amino acids, assisting in maintaining the relative water content. Up-regulation of the antioxidant system was obvious in AMF-inoculated plants, thereby mediating quick alleviation of oxidative effects of drought through elimination of ROS. In addition, AMF mediated up-regulation of the antioxidant system contributed to maintenance of redox homeostasis, leading to protection of major metabolic pathways, including photosynthesis, as observed in the present study. Total phenols increased due to AMF inoculation under both MD and SD conditions. The present study advocates the beneficial role of G. versiforme inoculation in maize against drought stress.

A survey of the incidence of dyslipidemia and its components in people over 20 years old in Ahvaz: A cohort study 2009-2014.

INTRODUCTION: Dyslipidemia is one of the major risk factors for atherosclerosis and the main cause of death in the developing and developed countries. In this cohort study, the incidence of dyslipidemia and its components have been studied among individuals over 20 years old in Ahvaz, southwest of Iran. MATERIALS AND METHODS: The Phase 1 of this study was conducted on the prevalence of Metabolic syndrome and its related factors by Diabetes Research Center in 2009. The target population included individuals over 20 years old who were selected using cluster sampling in Ahvaz health centers. In the second phase of the study, 5 years later in 2014, subjects were again recalled by health centers. Questionnaires, anthropometric indices and measurements were repeated similar to Phase 1. The blood sample was taken from everyone after 12h of fasting and then the factors (Chol, TG, HDL) were measured. RESULTS: The cumulative incidence of dyslipidemia obtained 60.7% (males 56.7% and females 64.8%). lipid profile {HDL, Triglycerides and Cholesterol had changes in their means(a significant reduction in the mean about 10, 22.25 and 13 units respectively) between baseline and 5 years later. Waist circumference here associated with incidence of high LDL and high chol, also sex and High LDL 'educational level and daily consumption of vegetables were associated with incidence of low HDL and dyslipidemia respectively. CONCLUSION: Age, educational level and daily intake of fruits and vegetables were effective variables on incidence of dyslipidemia. According to various studies, LDL and HDL levels in different regions of Iran, seems to be influenced by factors such as climate, nutrition and activity levels.

[Pancreatic neuroendocrine neoplasms]. / Neuroendokrine Neoplasien des Pankreas.

Pancreatic neuroendocrine neoplasms (NEN) account for 1-2% of all pancreatic neoplasms and represent a rare differential diagnosis. While some pancreatic NEN are hormonally active and exhibit endocrine activity associated with characteristic symptoms, the majority are hormonally inactive. Imaging techniques such as ultrasound, computed tomography (CT), magnetic resonance imaging (MRI) and positron emission tomography (PET) or as combined PET/CT play a crucial role in the initial diagnosis, therapy planning and control. Endoscopic ultrasound (EUS) and multiphase CT represent the reference methods for localization of the primary pancreatic tumor. Particularly in the evaluation of small liver lesions MRI is the method of choice. Somatostatin receptor scintigraphy and somatostatin receptor PET/CT are of particular value for whole body staging and special aspects of further therapy planning.

Introduction of a metabolic joint asymmetry score derived from conventional bone scintigraphy. A new tool to differentiate psoriatic from rheumatoid arthritis.

AIM: Clinical differentiation of psoriatic arthritis (PsA) and rheumatoid arthritis (rA) based on the pattern of joint involvement can be difficult; the frequent form of PsA with polyarthritis of the peripheral joints may sometime resemble rA. We investigated a metabolic joint asymmetry score (MJAS), reflecting the overall asymmetric joint involvement on conventional bone scintigraphy, for differentiating PsA from rA in patients presenting with peripheral polyarthritis. PATIENTS, METHODS: 106 patients (n = 61, PsA; n = 45, rA) with peripheral polyarthritis (≥ 5 joints) as well as 26 control subjects with no history of chronic joint disorders were analyzed. The intensity of articular 99mTc-MDP uptake in 40 peripheral joint pairs was scored regarding the bilateral difference of each joint based on a scale of 0-2 (no significant, moderate, and marked asymmetry, respectively). The patient's MJAS was defined as the sum of uptake difference scores of all joint pairs. The association of MJAS with the underlying condition (Psoriasis criteria, Revised Criteria of the ACR) was examined. RESULTS: 5280 peripheral joint pairs were investigated. There was no significant difference in the total number of involved joints in PsA 15.0 ± 8.2 versus rA 17.5 ± 8.8 patients (p = 0.132), but significantly less involvement in the control group (6.7 ± 5.0, p < 0.001). MJAS was markedly higher in PsA (17.0 ± 9.6) than in rA (4.8 ± 3.9, p < 0.001), and correlated with the total number of involved joints in PsA (r = 0.516, p < 0.001), but not in rA (r = 0.078, p = 0.380). The MJAS disparity between PsA and rA persisted after exclusion of the DIP joints (14.4 ± 7.7 vs. 4.4 ± 3.3; p<0.001). CONCLUSIONS: The new reproducible semi-quantification method for the asymmetry of metabolic joint involvement permits differentiation of psoriatic from rheumatoid peripheral arthritis with MJAS being markedly higher in patients with PsA as compared to rA patients. The score may offer an effective complementary tool for characterizing patients with peripheral polyarthritis.

Survival in patients with hepatocellular carcinoma treated with 90Y-microsphere radioembolization. Prediction by 18F-FDG PET.

AIM: This retrospective study aims to evaluate the predictive value of FDG PET/CT in patients with unresectable hepatocellular carcinoma (HCC) undergoing radioembolization with yttrium-90 labeled microspheres (RE). PATIENTS, METHODS: The study cohort comprised 33 patients who were treated with RE at our institution and underwent FDG PET/CT at baseline and four weeks after radioembolization. According to the baseline FDG metabolic status of the HCC lesions, patients were divided into two groups: FDG-negative (n = 12) and FDG-positive (n = 21) HCC. FDG-positive patients were further divided into early metabolic responders and non-responders according to the relative change in SUVmax of the treated lesions. Survival analyses were performed with the Kaplan-Meier method (log-rank test, p < 0.05). Multivariate analysis was performed to assess the influence of prognostic factors on overall survival (OS). RESULTS: FDG-negative patients had a significantly longer OS (13 months, 95%CI 7-19) than FDG-positive patients (9 months, 95%CI 7-11; p = 0.010). Among FDG-positive patients, metabolic responders survived significantly longer than metabolic non-responders (10 months, 95%CI 8-12 vs. 5 months, 95%CI 4-6; p = 0.003). From the other baseline factors (including performance status, hepatic tumour burden, presence of extra-hepatic disease, administered activity) only the BCLC stage had a significant impact on OS (p = 0.028). CONCLUSION: Pre- and post-therapeutic FDG PET independently predicts overall survival in patients with HCC undergoing radioembolization. Interestingly, early metabolic response seems to be assessable as early as four weeks post-treatment.

Can peptide receptor radionuclide therapy be safely applied in florid bone metastases? A pilot analysis of late stage osseous involvement.

AIM: Highly advanced metastatic bone disease with extensive osseous infiltration of neuroendocrine tumours (NET) may preclude patients from treatment with peptide receptor radionuclide therapy (PRRT) in concern about haematotoxicity. This study aims to assess the safety and efficacy of PRRT with 177Lu-octreotate in a patient cohort with this condition. PATIENTS, METHODS: 41 PRRT courses were performed in 11 patients with gastroenteropancreatic neuroendocrine tumours (GEP-NET) and florid bone metastases (severely advanced widespread metastatic bone disease). A mean activity of 6.95 GBq 177Lu-octreotate was administered per treatment cycle, aimed at four courses with standard intervals of 3 months. Haematological parameters were determined prior to each treatment course, in 2-4 weeks intervals between the courses, 8-12 weeks after the last course of PRRT and in 3 monthly intervals thereafter. Toxicity was recorded using Common Terminology Criteria for Adverse Events v3.0. Restaging was performed 3 months after termination of PRRT with CT/MRI and functional imaging (modified MDA criteria). RESULTS: Significant (grade III-IV), reversible haematotoxicity occurred in 4 (35%) patients and after 10 (24%) administrations. It either resolved spontaneously (1 patient) or was controlled by supportive measures (3 patients), such as blood transfusions (3 patients) or deferral of the subsequent therapy cycle (1 patient). Patients returned to baseline blood values within up to 23 months after termination of PRRT. The observed treatment response of bone metastases consisted of a partial response in 2, a minor response in 1, stable disease in 7, and progressive disease in 1 patient. Of the 4 patients with metastatic bone pain, 1 experienced complete and 3 partial resolution of symptoms within 3-10 weeks after commencement of PRRT. CONCLUSION: These preliminary data indicate that PRRT with 177Lu-octreotate can be safely applied even in florid bone metastases with extensive, severely advanced osseous replacement. The higher myelosuppression rate was not associated with serious complications and should not preclude patients from being treated and potentially experiencing remarkable treatment efficacy despite the very advanced stage.

Liver and lymph node metastases of prostate cancer visualized on post-therapy imaging after treatment with 188Re-HEDP / Metástasis hepáticas y linfáticas del cáncer de próstata en la imagen post-terapia con 188Re-HEDP

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Residual activity after radioembolization of liver tumours with 90Y resin microspheres. A safe calculation method.

UNLABELLED: The actual number of resin microspheres is approximately 30-60 times higher than glass microspheres per 3 GBq vial. Thus, radioembolization (RE) with resin microspheres exerts an embolization effect besides the radiation effect. This embolization effect can occasionally cause early back flow of the microspheres before application of the entire calculated dose. To avoid these adverse side effects, RE has to be terminated at an earlier time point. Measurement of the residual activity in the delivery box, which includes the v-vial, tube and catheter, to calculate the achieved target dose is often challenging. The aim of the current study was to establish a post-RE measurement method comparable to the glass microspheres method without unnecessary radiation exposure to the staff and risk of contamination. METHODS: Two different measurements were performed. First, total radioactivity in the shipping vial was measured in an ion chamber and then it was put in the delivery box and the radiation was measured from a 30 cm distance from the centre of the box with a dosimeter. The required radioactivity was then transferred to the v-vial, and the shipping vial was measured again. After that, the v-vial was measured from the same distance from the centre of the box with dosimeter. RESULTS: Altogether 62 times the shipping vial with different activities were measured with a significant positive correlation between the amount of the activity measured in the iron chamber and the radiation dose, measured with dosimeter (r² = 0.98; p< 0.001). There was also a strong positive correlation between these measurements of the v-vial (r² = 0.98; p< 0.001). CONCLUSION: With measurement of the residual activity in the delivery box using a dosimeter the percentage of the whole injected activity can be easily calculated. This facilitates the calculation of the actual, achieved target and non-target dose in those cases, where therapy had to be stopped because of eminent flow reversal or obstruction.

Early prediction of tumour response to PRRT. The sequential change of tumour-absorbed doses during treatment with 177Lu-octreotate.

UNLABELLED: [177Lu-DOTA0,Tyr3]-octreotate (177Lu-octreotate) in peptide receptor radionuclide therapy (PRRT) offers direct intra-therapeutic dosimetry. The aim of this study was to compare tumour and non-tumour parameters and assess intra-individual variations. PATIENTS, METHODS: Retrospective analysis of 53 consecutive PRRT treatment cycles (mean activity of 7.53 ± 0.46 GBq 177Lu-octreotate, intended four cycles at intervals of 10-14 weeks, standard nephroprotection) in 27 GEP NET patients. Extended planar dosimetry with serial whole-body imaging on selected, non-superimposed tumour and non-tumour regions; liver (LM), bone (BM), and other (OM) metastases. The per-cycle variation was compared with post-treatment response (CT/MRI three months post-treatment, modified SWOG criteria). RESULTS: Residence time in tumor lesions (133-147 h) exceeded that in kidneys (93 h). Tumour-to-kidney absorbed dose ratios ranged from 14 to 28 (LM, BM, OM). Intra-individual per-cycle dose variation was insignificant for kidneys, but significant for metastases (LM, BM, and OM; p < 0.05). The mean per-cycle decrease of tumour absorbed dose (ΔD/A0[%]) was linked to morphologic response after PRRT. A mean decrease of >20% was predictive of a partial or minor remission in all 11 evaluable patients, while absent significant dose reduction indicated stable or progressive disease in 4/5 patients. The dose decrease was unrelated to volume effects and also observed for BM. CONCLUSION: Besides confirmation of a favourable tumour-to-kidney parameter relation for 177Lu-octreotate, stepwise intra-lesional comparison seems to imply a prognostic impact of tumor dosimetry: The early per-cycle change ΔD/A0 between treatment cycles may predict the outcome after PRRT. Larger studies are needed to confirm this finding.

Association between troponin T and ICU mortality, a changing trend.

BACKGROUND: Initially elevated levels of troponin predict adverse outcomes in patients admitted to the intensive care unit (ICU). No research team has investigated the changes in concentration of cardiac troponin T (cTnT) during ICU stay and their association with patient outcome. OBJECTIVE: We investigated whether the change in cTnT levels during ICU stay could predict outcomes (death or survival). METHODS: In this cohort study, all patients admitted to the medical ICU (10 beds) from January to July 2008 were enrolled. Troponin levels were evaluated within the first 24 hours of ICU admission and on the fourth, seventh and 10th days after admission. RESULTS: The study population (135 patients) had a mean age of 60.9 ± 21.5 years. The outcome was significantly different with regard to normal or elevated cTnT concentrations on the first and seventh days of follow up (p = 0.03 and 0.023, respectively). This difference was non-significant for cTnT levels on the fourth and 10th days after admission (p = 0.69 and 0.78, respectively). The change in cTnT levels was not significantly different between the deceased and discharged patients (p = 0.4). CONCLUSION: Changes in cTnT levels during ICU stay did not show a significant trend (power: 0.26). Patients whose cTnT levels were increased on the first and seventh days of ICU stay had a worse survival, which could be associated with cardiac events on admission or at specific times during the stay in ICU.

Osseous metastases of gastro-enteropancreatic neuroendocrine tumours. Diagnostic value of intra-therapeutic 177Lu-octreotate imaging in comparison with bone scintigraphy.

AIM: Peptide receptor radionuclide therapy with 177Lu-octreotate is an effective treatment option for metastatic gastroenteropancreatic neuroendocrine tumors (GEP NET) and allows intratherapeutic imaging through a 177Lu-octreotate scan (LuS). The diagnostic value of this treatment scan is not yet established. This study aims to compare the sensitivity of LuS and bone scintigraphy (BS) regarding bone metastases and investigate potential implications of functional imaging results. PATIENTS, METHODS: We retrospectively analyzed 29 consecutive GEP NET patients with bone metastases and baseline BS treated with 177Lu-octreotate. A semi-quantitative scoring system was used for the comparative evaluation. Treatment outcome (time-to-progression of bone metastases) was correlated with the intra-individual imaging discrepancy (Kaplan-Meyer curves, log-rank test, p < 0.05). RESULTS: In 19 of 29 patients (65.5%) LuS was superior (LuS > BS), whereas in 10 patients (34.5%) both modalities were comparable. BS showed no additional (LuS-negative) metastatic bone lesions in our cohort. None of the investigated baseline characteristics was associated with imaging discrepancy. On the other hand, functional imaging discrepancy had no impact on treatment response (p = 0.43) or time-to-progression (p = 0.92). CONCLUSIONS: Intra-therapeutic 177Lu-octreotate imaging is superior over bone scintigraphy for detection of bone metastases in GEP NET. BS may help to distinguish osseous from non-osseous localization. The presence of an osteoblastic correlate in BS seems to have no impact on therapeutic outcome.

[SPECT-CT of the foot]. / SPECT-CT des Fußes.

UNLABELLED: During the last years SPECT-CT combining the high resolution of CT with high sensitivity of bone SPECT has gained widespread clinical application. Especially in complex anatomic situations, SPECT-CT allows to exactly define the area of the skeleton which is responsible for pain. This holds true especially in patients with foot diseases. CONCLUSION: Bone SPECT-CT gives additional useful information in arthrodesis, infection, accessory bones (sesamoids) as well as tarsal coalition.

Magnetic Resonance Measurement of Transient Shear Wave Propagation in a Viscoelastic Gel Cylinder.

A magnetic resonance measurement technique was developed to characterize the transient mechanical response of a gel cylinder subjected to angular acceleration. The technique employs tagged magnetic resonance imaging (MRI) synchronized to periodic impact excitation of a bulk specimen. The tagged MRI sequence provides, non-invasively, an array of distributed displacement and strain measurements with high spatial (here, 5 mm) and temporal (6 ms) resolution. The technique was validated on a cylindrical gelatin sample. Measured dynamic strain fields were compared to strain fields predicted using (1) a closed-form solution and (2) finite element simulation of shear waves in a three-parameter "standard" linear viscoelastic cylinder subjected to similar initial and boundary conditions. Material parameters used in the analyses were estimated from measurements made on the gelatin in a standard rheometer. The experimental results support the utility of tagged MRI for dynamic, non-invasive assays such as measurement of shear waves in brain tissue during angular acceleration of the skull. When applied in the inverse sense, the technique has potential for characterization of the mechanical behavior of gel biomaterials.

The National Registry of Veterans with amyotrophic lateral sclerosis.

BACKGROUND: The Department of Veterans Affairs (VA) Cooperative Studies Program has established a National Registry of Veterans with Amyotrophic Lateral Sclerosis (ALS). This article describes the objectives, methods, and sample involved in the registry. METHODS: United States military veterans with ALS were identified through national VA electronic medical record databases and nationwide publicity efforts for an enrollment period of 4 1/2 years. Diagnoses were confirmed by medical record reviews. Registrants were asked to participate in a DNA bank. Follow-up telephone interviews are conducted every 6 months to track participants' health status. RESULTS: As of September 30, 2007, 2,400 veterans had consented to participate in the registry, 2,068 were included after medical record review, 995 were still living and actively participating, and 1,573 consented to participate in the DNA bank. 979 participants had been enrolled in the registry for at least 1 year, 497 for at least 2 years, and 205 for at least 3 years. Fourteen studies have been approved to use registry data for epidemiological, observational, and interventional protocols. CONCLUSION: This registry has proven to be a successful model for identifying large numbers of patients with a relatively rare disease and enrolling them into multiple studies, including genetic protocols.

Skin biopsies demonstrate MPZ splicing abnormalities in Charcot-Marie-Tooth neuropathy 1B.

OBJECTIVE: To demonstrate that intronic mutations in the myelin protein zero (MPZ) cause Charcot-Marie-Tooth neuropathy 1B (CMT1B) by disrupting MPZ splicing. METHODS: We report a family with a T>G transversion at the invariant + 2 position in intron 4 of MPZ (c.614 + 2T>G) that abolishes 5' donor site recognition and is predicted to alter MPZ splicing. We obtained detailed clinical and neurophysiologic analysis of the family. We performed skin biopsies to investigate splicing abnormalities, MPZ protein levels, and localization in myelinated nerves. RESULTS: Patients developed a late onset neuropathy with minimally slow nerve conduction velocities. Skin biopsies confirmed the predicted skipping of exon 4 and downstream frameshift of the mutant MPZ. Quantitative immuno-EM demonstrated normal nerve MPZ levels, suggesting that the mutant MPZ was transported to compact myelin. CONCLUSIONS: Intronic mutations cause CMT1B by disrupting splicing and certain MPZ mutations may cause neuropathy by interacting with the wild type MPZ in the extracellular space of compact myelin.