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Efforts to develop and commercialize T cell immunotherapies would benefit from sensitive and easy-to-use assays to monitor target cell killing. We have developed an approach to selectively measure the death of target cells mediated by CAR/TCR-T cells or other types of effector cells. The method relies on the release of a HiBiT-tagged protein from target cells following cell lysis. Once released, HiBiT, an 11 a.a. peptide tag, binds to cell-impermeable Large BiT (LgBiT), a 17.6 kDa protein, to reconstitute NanoBiT Luciferase. In the presence of furimazine substrate, the luminescent signal is proportional to the amount of target cell killing, and cell lysis can be quantified at a single time point using Maximum Release and Spontaneous Release controls. The assay is compatible with a wide range of target cells per well (e.g. 100-500,000 cells/well), and low levels of Spontaneous Release allow the sensitive detection of low levels of target cell lysis (e.g. 1-5%). Using a modified format, target cell killing can be measured continuously in a bench-top luminometer for 24 hours or more. We demonstrate these approaches using commercially available CAR-T cells and target cells stably expressing a HiBiT-tagged protein via random integration of plasmid DNA or viral transduction. We also demonstrate the use of a thaw-and-use format to facilitate MOA-based lot release assays. The approach provides simple, non-radioactive assay formats for selectively monitoring target cell killing in mixed culture experiments. Citation Format: Brock Binkowski, Aileen Paguio, Christopher Eggers, Braeden Butler, Michael Beck, Frank Fan. A novel, bioluminescent assay for the selective detection of target cell killing in mixed cultures [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3181.

Purpose Recent studies have reported low reliability using the multistage cervical vertebral maturation (CVM) approach, raising concerns about the clinical usefulness of this method. The objective of the present study was to examine the reliability of a 2 stage CVM method. Materials and methods Thirteen orthodontic residents received standard trainings on the CVM methods (5 stage version), and classified the maturation status of 15 CVM radiographs representing all 5 stages of maturation. The radiographs were reevaluated 2 weeks later. The intra- and inter-rater reliability of the residents’ classifications were assessed by percentage agreement, Kendall's W test, weighted kappa (K w ) for the five stage method and simple kappa (K) for the pre vs. post mandibular growth peak phases (2-stage approach). The κ w values for the individual stages of the five stage method were also calculated. To assess percentage agreement and Kappa changes caused solely by category reduction (from 5 to 2), percentage agreement and simple Kappa statistics were also calculated for all other possible cut points (i.e. 1–2, 3–4 and 4–5). Results Based on 5 stages, the average intra-rater reliability showed 62% for exact agreement and 0.86 for κ w , the average inter-rater reliability showed 46.7% for exact agreement, 0.79 for both κ w and Kendall's W . The κ w for the intra-rater reliability of individual five stages ranged from .06 to .62 with the two highest at the stage 2–3 interface. Based on two stages (pre-post mandibular growth peak), the average intra-rater reliability showed 95% exact agreement and 0.82 for K The average inter-rater reliability showed 91% for exact agreement and 0.76 for K. For all other possible 2-stage cut points, the percentage agreement and Kappa measures were smaller than those from the cut point of pre and post peak growth. Conclusions The 2 stage CVM method has excellent reliability in differentiating pre and post peak mandibular growth phases. As such information is necessary for planning growth modification, the CVM method is a useful clinical tool and should be combined with other clinical assessments.

Aim To identify a means to objectively measure corneal clouding in patients with mucopolysaccharidosis in a prospective controlled clinical trial. Methods Corneal haze was assessed by slit lamp examination and measured using the densitometry programme of the Pentacam, a rotating Scheimpflug camera in 33 mucopolysaccharidoses (MPS) patients and 32 controls. Results Pentacam measurements were available in 31 right and 31 left eyes of 32 patients and in 32 left and right eyes of 32 subjects in the control group. Slit lamp findings correlated very well with corneal density measurements (Spearman correlation right eye (OD)/left eye (OS)=0.782/0.791). MPS patients had higher density units (median OD/OS=14.1/14.7) than control subjects (median OD/OS=6.7/6.9, p

AIM Prostate-specific membrane antigen (PSMA) is overexpressed in most prostate cancers (PCs). Here, we report our first experience using the Tc-labeled PSMA inhibitor MIP-1404 (Progenics Pharmaceuticals, Inc, Tarrytown, NY) in 60 patients with biochemically recurrent PC. METHODS Whole-body planar scintigraphy and SPECT/CT of the lower abdominal pelvic region of 60 patients with biochemical relapse of PC were analyzed retrospectively. In these subjects, an average dose of 733.1 ± 49.5 MBq (19.8 ± 1.3 mCi) Tc-labeled MIP-1404 was injected 4 to 5 hours prior to imaging. In addition to visual evaluation, SUVmax were determined in the tumor lesions using a previously developed protocol for quantitative SPECT/CT. RESULTS In 42 of 60 patients, Tc-MIP-1404-positive lesions could be detected (70%; 95% confidence interval [CI], 0.58-0.82). Twenty patients had Tc-MIP-1404-positive lymph nodes suggestive of metastasis, 14 patients had pathological uptake in the prostate region indicative of local recurrence, and for another 19 patients, there was tracer accumulation in the skeleton (n = 18) or lungs (n = 1). Detection rate was 91.4% (95% CI, 0.82-1) at prostate-specific antigen levels greater than 2 ng/mL and 40.0% (95% CI, 0.21-0.59) at lower prostate-specific antigen values (P < 0.01). Of the 60 patients, in total, 82 positive lesions were analyzed quantitatively. Average SUVmax of the lesions was 16.3 ± 21.6 with a range of 1.7 to 142.9. CONCLUSION Tc-labeled PSMA inhibitor MIP-1404 is a promising SPECT tracer for detection of locally recurrent or metastatic prostate cancer.

Background The therapy response of osseous metastases (OM) is commonly monitored by bone scintigraphies (BS). The aim of this study was to compare visual evaluation of changes in tracer uptake with quantitation in absolute units in OMs; 52 OMs from 19 patients who underwent BS with SPECT/CT at time points one and two (TP1/2) were analyzed retrospectively, with an average of 10.3 months between TP1 and 2. Tracer uptake in lesions was visually compared by two independent readers in both planar scintigraphies and SPECT/CT across both TPs and classified as regressive, stable, or progressive. Quantitative analysis was performed by measuring peak standardized uptake values (SUV). Based on quantitation, lesions were similarly classified as regressive (>30 % decrease), progressive (>30 % increase), or stable (rest). If available, uptake in reference regions in the lower thoracic or lumbar spine was used for normalization. Results In OMs at TP1 and TP2, mean SUVpeak (±SD) was found to be 20.4 (±20.8) and 16.4 (±11.5), respectively. For the reference region, mean SUVmean was 5.6 (±1.9) and 4.9 (±2.2). Agreement between quantitative and visual assessment was only moderate, with an average Cohen’s kappa of 0.42 for planar scintigraphy and 0.62 for SPECT/CT. Discrepancies occurred in between 11 and 22 of the 52 lesions, depending on the reader and whether planar or SPECT imaging was considered. Conclusions Compared to measuring uptake in absolute units, visual evaluation of skeletal scintigraphies for change in tumor metabolism yields inconsistent results in roughly one third of the cases.

WOS: 000377110800007, PubMed ID: 26908836, Background Mucopolysaccharidosis VII (MPS VII) is an ultra-rare disease characterised by the deficiency of beta-glucuronidase (GUS). Patients' phenotypes vary from severe forms with hydrops fetalis, skeletal dysplasia and mental retardation to milder forms with fewer manifestations and mild skeletal abnormalities. Accurate assessments on the frequency and clinical characteristics of the disease have been scarce. The aim of this study was to collect such data. Methods We have conducted a survey of physicians to document the medical history of patients with MPS VII. The survey included anonymous information on patient demographics, family history, mode of diagnosis, age of onset, signs and symptoms, severity, management, clinical features and natural progression of the disease. Results We collected information on 56 patients from 11 countries. Patients with MPS VII were classified based on their phenotype into three different groups: (1) neonatal non-immune hydrops fetalis (NIHF) (n=10), (2) Infantile or adolescent form with history of hydrops fetalis (n=13) and (3) Infantile or adolescent form without known hydrops fetalis (n=33). Thirteen patients with MPS VII who had the infantile form with history of hydrops fetalis and survived childhood, had a wide range of clinical manifestations from mild to severe. Five patients underwent bone marrow transplantation and one patient underwent enzyme replacement therapy with recombinant human GUS. Conclusions MPS VII is a pan-ethnic inherited lysosomal storage disease with considerable phenotypical heterogeneity. Most patients have short stature, skeletal dysplasia, hepatosplenomegaly, hernias, cardiac involvement, pulmonary insufficiency and cognitive impairment. In these respects it resembles MPS I and MPS II. In MPS VII, however, one unique and distinguishing clinical feature is the unexpectedly high proportion of patients (41%) that had a history of NIHF. Presence of NIHF does not, by itself, predict the eventual severity of the clinical course, if the patient survives infancy., Ultragenyx Pharmaceutical [16232], This work was supported in part by Ultragenyx Pharmaceutical grant number 16232.

We use time-resolved photoluminescence (TRPL) spectroscopy to unequivocally clarify the microscopic origin of the nanosecond free exciton photoluminescence rise in GaAs at low temperatures. In crucial distinction from previous work, we examine the TRPL of the GaAs free exciton second LO-phonon replica. This enables us to simultaneously monitor the unambiguous time evolution of the total exciton population and the cooling dynamics of the initially hot free exciton ensemble. We demonstrate by a model based on the Saha equation and the experimentally determined cooling behavior that the long-debated slow photoluminescence rise is caused by time-dependent shifts in the thermodynamic quasiequilibrium between free excitons and the uncorrelated electron-hole plasma.

The purposes of this study were to quantify bone damage associated with insertion of 2 types of miniscrew implants and to relate the amount of bone damage to monocortical plate thickness.Nondrilling (n = 28) and self-drilling (n = 28) miniscrew implants (6 × 1.6 mm, Dentaurum, Newtown, Pa), and pilot holes (n = 26) were placed bilaterally in the maxillae and the mandibles of 5 adult dogs immediately after death. Bone blocks were cut, bulk stained with 1% basic fuchsin, embedded in methyl methacrylate, sectioned, and mounted. Monocortical plate thickness was measured adjacent to the miniscrew implant insertion site. Damage amounts were quantified at distances of 0 to 0.5 mm (adjacent region) and 0.5 to 1 mm (distant region) from the bone-implant interface. Total fractional damaged area (%), fractional microcracked area (%), and fractional diffuse damaged area (%) were quantified by using standard histomorphometric methods.The mean monocortical plate thickness of the specimens from the mandible (2.2 mm) was significantly (P0.001) greater than that of the maxillary specimens (0.9 mm). In the mandible, the 3 damage parameters were greater with self-drilling miniscrew implants than with nondrilling miniscrew implants; however, there were no differences in the damage parameters in the maxilla.Bone damage accumulation is related to the type of miniscrew implant and the thickness of the bone.

We investigated the effect of light availability and soil moisture on growth and biomass partitioning of Norway spruce and European beech seedlings in a three (light availability levels) × two (soil moisture levels) factorial greenhouse experiment. The effects of factor levels on allocation to biomass compartments were analyzed using ANCOVA. As plant allocation patterns are size-dependent, tree size was used as a covariate. In both tree species, growth and biomass allocation to above and belowground plant components were affected by light availability. European beech showed a distinct increase in allocation to leaves, stem and branch biomass at the expense of fine and coarse roots with decreasing light availability. For Norway spruce, only allocation to stem biomass increased and allocation to fine root biomass decreased under low light. To drought a significant increase of the percentage of belowground compartments was found for European beech but not for Norway spruce. Overall, European beech seedlings were more plastic than Norway spruce seedlings. European beech seedlings appear better able to adjust biomass partitioning to resource availability. In contrast Norway spruce responded languidly. Our results indicate that biomass partitioning is not only driven by ontogeny, and thus tree size, but is environmentally determined to a substantial degree. A possible explanation for this divergence from other results on the role of ontogeny in biomass partitioning may be that seedling plasticity in response to limited resources declines with increasing age and/or time of exposure to the limited resources.

Introduction Comprehension of informed consent information has been problematic. The purposes of this study were to evaluate the effectiveness of a shortened explanation of an established consent method and whether customized slide shows improve the understanding of the risks and limitations of orthodontic treatment. Methods Slide shows for each of the 80 subject-parent pairs included the most common core elements, up to 4 patient-specific custom elements, and other general elements. Group A heard a presentation of the treatment plan and the informed consent. Group B did not hear the presentation of the informed consent. All subjects read the consent form, viewed the customized slide show, and completed an interview with structured questions, 2 literacy tests, and a questionnaire. The interviews were scored for the percentages of correct recall and comprehension responses. Three informed consent domains were examined: treatment, risk, and responsibility. These groups were compared with a previous study group, group C, which received the modified consent and the standard slide show. Results No significant differences existed between groups A, B, and C for any sociodemographic variables. Children in group A scored significantly higher than did those in group B on risk recall and in group C on overall comprehension, risk recall and comprehension, and general risks and limitations questions. Children in group B scored significantly higher than did those in group C on overall comprehension, treatment recall, and risk recall. Elements presented first in the slide show scored better than those presented later. Conclusions This study suggested little advantage of a verbal review of the consent (except for patients for risk) when other means of review such as the customized slide show were included. Regression analysis suggested that patients understood best the elements presented first in the informed consent slide show. Consequently, the most important information should be presented first to patients, and any information provided beyond the first 7 points should be given as supplemental take-home material.

We study some relationships between holomorphic motions, continuous motions, and monodromy. We also study extensions of holomorphic motions over Riemann surfaces and characterize the extendability of holomorphic motions over some planar regions in terms of monodromy.

Zusammenfassung Zur Entwicklung neuer Medikamente für seltene Erkrankungen (Orphan Drugs) müssen auf Grund der geringen Zahl an Probanden andere Studien-Designs als das konventionelle randomisierte, doppelblinde, Plazebo-kontrollierte Protokoll zur Anwendung kommen. Die europäische Gesundheitsbehörde EMA unterstützt pharmazeutische Firmen bei der Entwicklung von Orphan Drugs durch erleichterte Zulassungsbedingungen. Der erhöhte Entwicklungsaufwand führt zu hohen Medikamentenpreisen, die den Patienten in vielen Ländern nicht erstattet werden.

Purpose The Mucopolysaccharidosis Type I (MPS I) Registry is an international observational database that tracks the natural history and the outcomes of patients with MPS I. The Registry was a regulatory requirement following the approval of laronidase enzyme replacement therapy for MPS I in 2003. All data are collected voluntarily after informed consent from the patient or family. Data are checked through queries, monthly reviews, and electronic audits to identify missing, inconsistent, or invalid data. This analysis sought to determine overall data accuracy in the Registry through source document verification (SDV). Methods Two phases of SDV were performed. In each phase, Registry data were compared against source documents at sites in Europe, Latin America, and North America. Three patients were randomly selected for SDV at each of the selected sites among all patients enrolled ≥18 months and ever receiving laronidase. Key parameters central to MPS I and its treatment were examined from the baseline and the last available assessments. Results Results indicate an overall source-to-database error rate in the MPS I Registry of 2.7% (47 discrepancies out of 1715 items; 95% confidence interval [2.2%, 3.5%]) in Phase 1 and 3.7% (64 discrepancies out of 1732 items; 95% confidence interval [2.9%, 4.7%]) in Phase 2. No systematic errors were found. Conclusions The overall error rates in both phases of SDV demonstrate acceptable data accuracy in the MPS I Registry within the data fields that were assessed. Copyright © 2011 John Wiley & Sons, Ltd.

Background: Neuropsychiatric symptoms (NPS), mainly cognitive deficits up to dementia and depressive syndromes have been described repeatedly in Fabry disease (FD). However, examinations regarding the pattern, extent, and frequency of the NPS in FD are still lacking. Moreover, the relationship between NPS and brain structural alterations in FD is unknown. The aim of this study was 1) to characterize NPS in a relatively large cohort of adult subjects with FD, and 2) to explore the association of cognitive performance and depressive syndromes with the FD-typical brain structural findings. Methods: Twenty-five Fabry patients (age 36.5 ± 11.0) with mild to moderate disease involvement and 20 age, gender-, and education-matched healthy controls were extensively studied by neuropsychiatric assessment, structural magnetic resonance imaging, magnetic resonance angiography, and diffusion-tensor imaging. Results: Patients with FD showed deficits only in the attention domain. Clinically relevant depressive syndromes were noted in 60% of the patients. The subgroup of patients with markedly elevated volumes of white matter lesions (not associated with actual stroke; n = 7) showed slightly more learning and memory deficits, but no higher depression rate compared to less affected patients. Conclusions: Against the prevailing assumption, Fabry patients, even those with marked brain structural alterations, showed only mild cognitive deficits. The high frequency of depression in FD is likely to be related to the burden of this chronic multiorganic hereditary disease, but not to the FD-typical brain structural alterations. Longitudinal studies are necessary to clear, if the mild cognitive deficits in FD might precede clinically relevant cognitive decline.

Introduction Computer-based smile esthetic surveys based on slider technology allow more precise control of variables and the possibility of obtaining continuous data. Variations in the perception of smiles from different facial perspectives have not been resolved. The objective of this study was to quantify the ideal and the range of acceptable values for smile variables judged by laypersons from a full-face perspective for comparison with lower-face data. Methods Mirrored and symmetric male and female full faces previously determined by peers to be of average attractiveness were used. Ninety-six laypersons judged these smile variables: smile arc, buccal corridor fill, maxillary gingival display, maxillary midline to face, maxillary to mandibular midline discrepancy, overbite, central incisor gingival margin discrepancy, maxillary anterior gingival height discrepancy, incisal edge discrepancy, and cant. The judges manipulated the variables using adjustable image technology that allowed the variable to morph and appear continuous on a computer monitor. Medians for each smile variable were compiled, and the Fleiss-Cohen weighted kappa statistic was calculated to measure reliability. Multiple randomization tests with adjusted P values were used to compare these data with those for lower-face views. Results Reliability ranged from 0.25 for ideal overbite to 0.60 for upper midline to face, except for upper and lower buccal corridor limits, which each had a kappa value near 0. There were no statistically significant differences between the ratings of male and female raters. The following variables showed statistically and clinically significant differences (>1 mm) when compared with the lower-face view: ideal smile arc, ideal buccal corridor, maximum gingival display, upper to lower midline, and occlusal cant. Although the smile arc values differed because of model lip curvature variations, the principle of tracking the curve of the lower lip was confirmed. For the full-face view, the raters preferred less maximum gingival display, less buccal corridor, more upper to lower midline discrepancy, and less cant of the occlusal plane. Conclusions Reliability was fair to moderate with the exception of the buccal corridor limits. Most variables showed no clinically meaningful differences from the lower-face view. The acceptable range was quite large for most variables. Detailed knowledge of the ideal values of the various variables is important and can be incorporated into orthodontic treatment to produce an optimal esthetic smile.

Helm NB, Padala S, Beck FM, D’Atri AM, Huja SS. Short-term zoledronic acid reduces trabecular bone remodeling in dogs. Eur J Oral Sci 2010; 118: 460–465. © 2010 Eur J Oral Sci The effects of zoledronic acid, a bisphosphonate, on the trabecular bone remodeling (TBR) of the mandibular condyle are unknown. The objectives of this study were to quantify and compare TBR in the mandibular condyle and vertebrae of 2- to 3-yr-old dogs and to evaluate the effects of short-term zoledronic acid on TBR. Bone samples from two, age-matched groups of dogs [seven dogs were given no treatment (NT group) and seven dogs were treated with four total infusions of zoledronic acid administered monthly (ZOL group)] were analyzed using histomorphometry. Trabecular bone remodeling and microarchitecture were quantified and analyzed statistically. Physiologic TBR, quantified in the NT group, was significantly higher (more than sixfold) in the vertebrae than in the mandibular condyle. Trabecular bone remodeling in the vertebrae of dogs of the ZOL group was 96% lower than in dogs of the NT group. By contrast, TBR in the mandibular condyle of dogs in the ZOL group was statistically equivalent to that of dogs in the NT group. Our results demonstrate that the physiological TBR in aged dogs is vastly different in the mandibular condyle compared to that in the vertebra. A higher level of physiologic TBR in the vertebra than in the mandibular condyle results in greater reduction of TBR in response to short-term treatment with zoledronic acid.

Immune costimulatory receptors synergize with TCR-CD3 signaling to promote cell cycle progression, cytokine production, T cell survival and effector functions. They belong to immunoglobulin super-family (CD28, ICOS) and tumor necrosis factor receptor (TNFR) superfamily (4-1BB, OX40, CD27, CD40, GITR, DR3, and HVEM). Preclinical and clinical evidence shows that agonist antibodies against costimulatory receptors can stimulate antitumor immunity and are emerging as a promising approach for cancer immunotherapies. Furthermore, FcγR engagement, especially FcγRIIB due to its bioavailability, has been shown to provide a crosslinking scaffold for the antibody IgG to facilitate TNFR clustering/activation and become requisites for some agonist antibody activities. Here, we report the development of a suite of cell-based reporter bioassays to quantitatively measure the potencies of therapeutic antibodies designed to target costimulatory receptors. For this, we developed engineered effector cells that express each costimulatory receptor and a luciferase reporter driven by response element that specifically responds to costimulatory receptor signaling. We also developed a FcγR-expressing cell line to evaluate the involvement of FcγR in agonist antibody-mediated costimulatory signaling. The assays are specific for each costimulatory receptor, and can be used to detect both agonistic and blocking activities for the antibodies. When co-incubated with the engineered FcγR-expressing cells, they are able to show the Fcγ receptor-dependent or independent agonist activities. In summary, these reporter bioassays can serve as powerful tools in immunotherapy drug development for antibody screening and potency determination. Citation Format: Jun Wang, Michael Beck, Jamison Grailer, Pete Stecha, Jim Hartnett, Frank Fan, Mei Cong, Zhi-Jie Jey Cheng. Cell-based reporter bioassays to evaluate the Fc gamma receptor-dependent agonistic activities for therapeutic antibodies targeting immune costimulatory receptors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2732.

Background: Fabry disease is associated with cardiomyopathy, early-onset stroke, and progressive renal failure, and other features. No markers predict multisystemic disease progression. The hypothesis of the current study is to assess a clinically relevant marker for Fabry disease using polymorphic genotyping of a marker that has been shown to be involved in interrelated cardiac, vascular, and renal abnormalities in patients not affected by Fabry disease. The paraoxonase (PON1) polymorphisms, Leu55Met and Gln192Arg, modulate intima-medial artery thickening, prognosis of cardiovascular stroke, and renal failure in other diseases. Methods: PON1 polymorphisms were ascertained. The Mainz Severity Score Index (MSSI) for Fabry disease was calculated. Local Institutional Review Board approval was received. Results: 104 patients (58 female) and 46 controls (23 female) were evaluated. There was a significant difference (p = 0.04) in PON55LL (42.3%) among patients as compared to controls (21.7%) but none in PON192 genotypes. PON55 variant (MM) was correlated with severe MSSI renal sub-scores (p < 0.001) also when age-adjusted but not with cardiac, neurological, or general sub-scores. PON55LL genotype, correlated with higher PON1 activity, had lowest α-galactosidase-A activity (n = 45). Conclusion: There was no combined effect of PON55-PON192 polymorphisms. PON55LL was more common among patients. PON55MM genotype was correlated with non-mild renal sub-scores. However, sample size needs to be enlarged.

Because orphan drug regulations encouraged development of drugs for rare disorders by granting marketing exclusivity for many years and other commercial benefits, treatment has become achievable for a few lysosomal storage disorders also. The presently available therapies, however, are not able to address all aspects of these multisystemic disorders and do not cure the patient. Therefore, there is a need for producing new drugs that are based on known pathophysiological mechanisms, such as enzyme replacement or inhibition of substrate synthesis, or which use new approaches to prevent the build-up of storage material.New compounds that are being designed by different pharmaceutical companies can be divided in two groups, enzyme targeted and substrate targeted drugs. Enzyme targeted drugs include substances that modify the enzyme to make it more accessible to organs such as the bone or the brain, enhance enzyme activity (chaperones) or activate enzyme synthesis by small molecules that induce read-through of premature stop codons of genes that bear a nonsense mutation. To the group of substrate targeted drugs belong substances that inhibit the synthesis or modify the structure of the substrate (substrate deprivation or substrate optimization, respectively). For this review, a literature research has been undertaken that covers the years 1968 - 2010.The reader of this paper will get an overview of drugs for lysosomal storage disorders that are on the market or are under development. This will help in the understanding of the pathophysiological mechanisms that underlie these rare metabolic diseases. In addition, the reader should realize that the increasing number of these very expensive drugs may lead to significant consequences for the health economic system.In the last years, the interest of scientists and pharmaceutical companies in lysosomal storage disorders has increased dramatically, leading to the production of a rising number of different drugs. These drugs act at several stages of the pathophysiological cascade, for example, at the level of the substrate (substrate deprivation) or of the enzyme (enzyme enhancement). The presently available treatments are not able to address all clinical manifestations of these disorders, and it will still take a long time until new drugs are developed that are capable of curing the patients.

The mucopolysaccharidoses (MPS) are a group of lysosomal storage disorders characterized by a wide variation in symptoms and progression rates. Each of the MPS types is caused by a deficiency in one of the enzymes involved in the catabolic pathway of glycosaminoglycans. MPS are usually suspected on the basis of the concomitant presence of several typical features of the disease, such as short stature, coarse facial features, loss of hearing, bone deformities, joint stiffness, organomegaly, respiratory and cardiovascular complications, and, in some cases, developmental delay. Very reduced or absent enzyme activity is required to confirm an MPS diagnosis. In the past, treatment of MPS was limited to the management of individual symptoms of the diseases. Currently, specific therapies such as hematopoietic stem cell transplantation and enzyme replacement therapy have improved the outlook for many MPS patients. To monitor disease progression and the impact of therapy in these patients, there is need for a validated scoring system for evaluating disease severity in MPS. A scoring system should take into account all aspects of MPS, particularly quality of life and functioning of arms, hands and legs and related mobility, endurance, self care and social functioning. This paper discusses several of the available scoring systems for bone/skeletal disorders and their potential usefulness in patients with MPS.

Background: Fabry disease is an X-chromosomally inherited lysosomal storage disorder leading to accumulation of glycosphingolipids, mainly globotriaosylceramide (ceramide-trihexoside, Gb3). Concentrations of Gb3 in plasma and urine have been used to diagnose Fabry disease and to monitor enzyme replacement therapy with recombinant α-galactosidase. Methods: Gb3 was purified from plasma or urine by combined liquid extraction/protein precipitation and solid-phase extraction, and was detected by flow-injection analysis electrospray mass spectrometry (MS) using multi-reaction-monitoring. Calibration was performed via standard addition using C17-Gb3 as internal standard. The most abundant isoforms were monitored for calculation of total Gb3. Results: A MS-based assay for quantification of Gb3 in plasma and urine was established and validated. Intra- and inter-assay coefficient of variation (CV) of the method were ≤12%. However, at low concentrations the CV was 16%. The linear range covers roughly two orders of magnitude, down to 0.54 mg/L in plasma and 0.07 mg/L in urine. Careful adjustment of tuning parameters was necessary to obtain identical isoform intensities and quantitative results on different mass spectrometers. Gb3 concentrations in healthy controls were Conclusions: An improved MS protocol for Gb3 quantification has been developed, validated, and shown to be suitable for diagnosis and monitoring of Fabry patients. Clin Chem Lab Med 2010;48:189–98.

Microcomputed tomography (micro-CT) is increasingly being used to analyze the three-dimensional structure and architecture of microvascular networks. Therefore we have evaluated a micro-CT analysis of VEGF-induced vessel ingrowth into a porous polyurethane scaffold through comparison with analyses by CD31 immunohistochemistry, vascular perfusion by intravital Lycopersicon esculentum lectin perfusion and vascular corrosion casting. Micro-CT scanning found a similar level of vascularisation within the VEGF treated scaffolds to that determined by the other analytical methods. However, although the relative increase in vascularisation (17 fold above PBS controls p0.05) induced by VEGF determined by micro-CT was similar to the perfusion based analyses (20.1 and 10.4 fold for lectin perfusion and vascular corrosion respectively p0.05), it differed substantially from that determined by CD31 immunohistochemistry (3.2 fold p0.05). This difference was due to a large proportion of unperfused vessels in the PBS control that were not present in the VEGF group. The increase in perfusion probably resulted in part from an increase in average vessel diameter. Though this increase was detected by micro-CT, the actual diameters were overestimated by 60-90% most likely as a consequence of a merging effect for juxtaposed vessels. Thus whilst micro-CT gives an accurate three-dimensional quantification of the VEGF-induced increase in perfused vessels, resolution needs to be maximized for accurate sizing of a microvascular network's components.

Morbus Fabry ist eine erbliche lysosomale Speicherkrankheit, die durch einen Mangel an α-Galaktosidase A (α-Gal A) verursacht wird. Hierdurch sammeln sich Stoffwechselzwischenprodukte in unterschiedlichen Zelltypen an und beeintrachtigen die Funktion zahlreicher Organe. Mit progredienter Hamaturie, Albuminurie und Abnahme der glomerularen Filtrationsrate (GFR) sind die Nieren besonders schwer betroffen. Bisher ist die Substitution des fehlenden Enzyms die einzige Moglichkeit zur Behandlung der Erkrankung. Zwei Formen sind fur den Einsatz bei Patienten mit Morbus Fabry zugelassen: Agalsidase alfa und Agalsidase beta. In mehreren Studien bewirkten beide Enzyme nach Einleitung der Therapie eine Stabilisierung der Nierenfunktion, evaluiert mittels eGFR, Kreatininclearance und Serumkreatinin. Die Prognose ist offenbar deutlich besser fur Patienten mit milder oder mittelgradiger Niereninsuffizienz zum Zeitpunkt des Einsetzens der Enzymersatztherapie (ERT). Aus diesem Grund ist eine fruhestmogliche Enzymsubstitution obligatorisch. Des Weiteren wurde eine dosisabhangige Reduktion der Globotriaosylceramide intrazellular und im Urin nachgewiesen. Unbehandelt kommt es zu einer Abnahme der eGFR von ca. 12 ml/min/Jahr. Obwohl eine durchgehende Stabilisierung der Nierenfunktion durch die ERT mehrmals gezeigt wurde, konnte bisher eine sichere antiproteinurische Wirkung der ERT nicht bewiesen werden. Ferner sind renoprotektive Medikamente bisher in den ERT-Studien nicht konsequent eingesetzt worden. In jedem Fall sind weitere Studien zur Beurteilung der langfristigen ERT-Wirksamkeit auf Morbiditat und Mortalitat der Patienten mit Morbus Fabry notwendig.

Although Fabry disease is X linked and considered to affect primarily male hemizygotes, female heterozygotes may experience all the signs and symptoms of this metabolic disorder. This prospective, single-center, open-label, clinical trial was performed to evaluate the long-term response of female patients with Fabry disease to enzyme replacement therapy.Symptomatic women (average age = 47 years) enrolled in this 4-year study were treated with agalsidase alfa (Replagal, Shire HGT, Inc.) at a dose of 0.2 mg/kg, every other week for 4 years (N = 36). Clinical and biochemical assessments were conducted at 12-month intervals.The Mainz Severity Score Index, a measure of total disease burden, was significantly reduced after 12 months (P0.01) of treatment and continuously improved over 4 years. Brief Pain Inventory "pain at its worst" score was reduced from 4.6 +/- 2.9 at baseline to 3.3 +/- 2.9 after 12 months (P = 0.001) and remained reduced through 4 years. Mean left-ventricular mass decreased from 89.4 +/- 29.3(2.7) g/m at baseline to 66.5 +/- 29.3(2.7) g/m after 12 months (P0.001) and remained reduced through 4 years. Average kidney function (estimated glomerular filtration rate and proteinuria) remained constant during the study. No safety issues were identified.Long-term agalsidase alfa is effective and was well tolerated in women with Fabry disease.

In male patients with Fabry disease, an X-linked disorder of glycosphingolipid metabolism caused by deficient activity of the lysosomal enzyme alpha-galactosidase A, kidney dysfunction becomes apparent by the third decade of life and invariably progresses to ESRD without treatment. Here, we summarize the effects of agalsidase alfa on kidney function from three prospective, randomized, placebo-controlled trials and their open-label extension studies involving 108 adult male patients. The mean baseline GFR among 54 nonhyperfiltrating patients (measured GFR135 ml/min per 1.73 m(2)) treated with placebo was 85.4 +/- 29.6 ml/min per 1.73 m(2); during 6 mo of placebo, the mean annualized rate of change in GFR was -7.0 +/- 32.9 ml/min per 1.73 m(2). Among 85 nonhyperfiltrating patients treated with agalsidase alfa, the annualized rate of change was -2.9 +/- 8.7 ml/min per 1.73 m(2). Treatment with agalsidase alfa did not affect proteinuria. Multivariate analysis revealed that GFR and proteinuria category (1 oror = 1 g/d) at baseline significantly predicted the rate of decline of GFR during treatment. This summary represents the largest group of male patients who had Fabry disease and for whom the effects of enzyme replacement therapy on kidney function have been studied. These data suggest that agalsidase alfa may stabilize kidney function in these patients.

Neurologic hallmarks of Fabry disease (FD) include small fiber neuropathy as well as cerebral micro- and macroangiopathy with premature stroke. Cranial MRI shows progressive white matter lesions (WML) at an early age, increased signal intensity in the pulvinar, and tortuosity and dilatation of the larger vessels. To unravel the most promising imaging tool for the detection of CNS involvement in FD we compared the diagnostic utility of the different MR imaging findings.Twenty-five clinically affected patients with FD (age 36.5 +/- 11.0) and 20 age-matched controls were investigated by structural MRI, MR angiography, and diffusion tensor imaging (DTI). Individual WML volumes, global mean diffusivity (MD), and mean cerebral artery diameters were determined.Using receiver operating characteristic analyses, enlarged diameters of the following cerebral arteries significantly separated patients with FD from controls: middle cerebral artery: area under curve (AUC) = 0.75, p = 0.005; posterior cerebral artery: AUC = 0.69, p = 0.041; carotid artery: 0.69, p = 0.041; basilar artery: AUC = 0.96, p0.0005. A total of 87% of the individuals were correctly classified by basilar artery diameters (sensitivity 95%, specificity 83%). WML volumes and global MD values did not significantly separate patients from controls.With an accuracy of 87%, basilar artery diameters were superior to all other MR measures for separating patients with Fabry disease (FD) from controls. Future studies should adopt basilar artery measurements for early detection and monitoring of brain involvement in FD. Moreover, further investigations should reveal if the dilated vasculopathy in FD could be a screening marker to detect FD in a cohort of other cerebrovascular diseases, especially in cryptogenic stroke.

Fabry disease is an X-linked disease caused by the deficiency of the lysosomal enzyme alpha-galactosidase A. The lack of enzyme activity leads to progressive depositions of undegraded glycolipids in a great number of organs, resulting in a multisystemic disorder.Enzyme replacement therapy is available for patients with Fabry disease. Two enzyme preparations have been approved in the European Union by the European Agency for the Evaluation of Medicinal Products (EMEA): agalsidase beta (Fabrazyme, Genzyme Corporation), produced in Chinese hamster ovary cells; and agalsidase alfa (Replagal, Shire Human Genetic Therapies, Inc.), produced in human cell lines.This review details the latest reports regarding the clinical efficacy and tolerability of agalsidase alfa in patients with Fabry disease.Agalsidase alfa was shown to be effective in treating pain and in reducing heart size in patients with Fabry disease, to stabilize kidney function and to improve hearing, sweating and quality of life. It is able slow down progression of renal failure in patients with end-stage renal disease.

The objective of this study was to evaluate the long-term clinical benefits and safety of recombinant human arylsulfatase B (rhASB) treatment of mucopolysaccharidosis type VI (MPS VI: Maroteaux-Lamy syndrome), a lysosomal storage disease. Fifty-six patients derived from 3 clinical studies were followed in open-label extension studies for a total period of 97–260 Weeks. All patients received weekly infusions of rhASB at 1mg/kg. Efficacy was evaluated by (1) distance walked in a 12-minute walk test (12MWT) or 6-minute walk test (6MWT), (2) stairs climbed in the 3-minute stair climb (3MSC), and (3) reduction in urinary glycosaminoglycans (GAG). Safety was evaluated by compliance, adverse event (AE) reporting and adherence to treatment. Results: A significant reduction in urinary GAG (71–79%) was sustained. For the 12MWT, subjects in Phase 2 showed improvement of 255±191m (mean±SD) at Week 144; those in Phase 3 Extension demonstrated improvement from study baseline of 183±26m (mean± SE) in the rhASB/rhASB group at Week 96 and from treatment baseline (Week 24) of 117±25m in the placebo/rhASB group. The Phase 1/2 6MWT and the 3MSC from Phase 2 and 3 also showed sustained improvements through the final study measurements. Compliance was 98% overall. Only 560 of 4121 reported AEs (14%) were related to treatment with only 10 of 560 (2%) described as severe. Conclusion: rhASB treatment up to 5 years results in sustained improvements in endurance and has an acceptable safety profile.

Purpose: Hunter syndrome (Mucopolysaccharidosis II) is a rare, X-linked disorder of glycosaminoglycan metabolism. It is caused by a deficiency in the lysosomal enzyme iduronate-2-sulfatase, and in affected patients glycosaminoglycan accumulates in lysosomes of various tissues and organs and contributes to the pathophysiology of Hunter syndrome. The Hunter Outcome Survey (HOS) was established to better describe the natural history of this disorder and to evaluate the long-term effect of enzyme replacement therapy. Methods: HOS is an international, multicenter, long-term observational survey that will collect data on participating patients with a confirmed diagnosis of Hunter syndrome. Data will be collected during regular physician examinations and entered into an electronic database. Examples of observations include vital signs, laboratory values, signs and symptoms of organ involvement, and the results of selected functional tests (e.g., audiometry, echocardiogram, joint mobility, etc.). Results: As of May 15, 2007, 263 patients from 16 countries have enrolled in HOS; 24% of these patients were currently being treated with enzyme replacement therapy. The median age at enrollment was 12.2 years. The median age of onset of symptoms and diagnosis of Hunter syndrome were 1.5 and 3.5 years, respectively. Otitis media and abdominal hernia were the earliest presenting symptoms. Facial dysmorphism and hepatosplenomegaly were demonstrated by 95% and 89% of patients, respectively. Conclusions: HOS will be a valuable resource for enhancing the understanding of Hunter syndrome and will provide important information about the natural history of the disease and the role of enzyme replacement therapy in its treatment. Patients and their physicians should be encouraged to participate.

Aim: Fabry disease (Fabry) is a rare X-linked disorder caused by a deficiency of the lysosomal enzyme α-galactosidase A. The progressive accumulation of the major substrate, globotriaosylceramide, leads to renal dysfunction and hypertrophic cardiomyopathy, which are reported to become apparent in the third decade. This study was performed to determine if signs of cardiac manifestations of Fabry are seen in younger Fabry patients. Methods: Twenty children and adolescents of ≤18 years of age with confirmed Fabry underwent a standard 12-lead electrocardiogram (ECG), a 2-h Holter ECG, blood pressure measurements and a two-dimensional echocardiogram. Follow-up examinations were conducted for 14 patients after a mean interval of 25.9 months. Results: At baseline, mean left ventricular mass (LVM) indexed to height (LVM/h2.7) was 45.0 ± 2.3 and 47.0 ± 3.4 g/m2.7 in boys (n = 8) and girls (n = 12), respectively, and all patients had LVM/h2.7 of >75th percentile of that in healthy controls. After a mean 26-month follow-up, 12 out of 14 patients (85.7%) showed a mean increase of 7.5 ± 3.2 g/m2.7 in LVM/h2.7. Heart rate variability (HRV) analyses revealed that male, but not female, Fabry patients had significantly reduced HRV, reflecting a reduction in parasympathetic stimulation of the heart (p < 0.05). Conclusion: Cardiac involvement in children with Fabry is frequent and may progress even at young age.

Fabry disease, an X-linked inborn error of metabolism, is characterized by multi-organ involvement including cardiac signs of left ventricular hypertrophy and abnormal intima-medial (IMT) thickening of arteries, progressive renal failure, neurological involvement, and more. The vitamin D receptor (VDR) and an enzyme producing vitamin D3 result in an autocrine loop with direct effects on blood vessels. The purpose of this study is to assess VDR polymorphisms (BsmI, FokI, ApaI, and TaqI) relative to clinically important disease parameters using a disease-specific severity score (MSSI) and haplotype analysis. There were statistically significant differences between females (43% of 74 patients) and males in MSSI total scores, and in general and neurologic sub-scores. There appears to be a protective effect of the TaqI tt genotype so that there were significantly lower scores in clinical categories between those with the tt genotype versus those with the TT genotype. Multivariate models of haplotypes with MSSI scores reveal that T-A-f-B and t-a-F-b haplotypes of the VDR gene polymorphisms are significantly associated with variation in the Fabry phenotype. Despite the limitations of using the MSSI score as a clinical correlate, these results are provocative and further studies in larger cohorts with more males are recommended.

Mucopolysaccharidosis type II (MPS II; Hunter syndrome) is a rare X-linked recessive disease caused by deficiency of the lysosomal enzyme iduronate-2-sulphatase, leading to progressive accumulation of glycosaminoglycans in nearly all cell types, tissues and organs. Clinical manifestations include severe airway obstruction, skeletal deformities, cardiomyopathy and, in most patients, neurological decline. Death usually occurs in the second decade of life, although some patients with less severe disease have survived into their fifth or sixth decade. Until recently, there has been no effective therapy for MPS II, and care has been palliative. Enzyme replacement therapy (ERT) with recombinant human iduronate-2-sulphatase (idursulfase), however, has now been introduced. Weekly intravenous infusions of idursulfase have been shown to improve many of the signs and symptoms and overall wellbeing in patients with MPS II. This paper provides an overview of the clinical manifestations, diagnosis and symptomatic management of patients with MPS II and provides recommendations for the use of ERT. The issue of treating very young patients and those with CNS involvement is also discussed. ERT with idursulfase has the potential to benefit many patients with MPS II, especially if started early in the course of the disease.

This multicenter, open-label study evaluated pharmacokinetics, pharmacodynamics, and safety of agalsidase alpha in pediatric compared with adult patients with Fabry disease. The pharmacokinetic parameters of pediatric patients (19 boys, 5 girls, 6-18 years old; mean age, 11.8 years) were compared to those of adult male and female patients who participated in other clinical studies. All patients received agalsidase alpha at a dose of 0.2 mg/kg infused over 40 minutes every other week. Agalsidase alpha exhibited a biphasic serum elimination profile with a maximum serum concentration at the end of the 40-minute infusion

The different types (I–VII) of mucopolysaccharidoses (MPS), a hereditary lysosomal storage disorder where the enzyme defect leads to glycosaminoglycan deposits in different tissues, present with characteristic lesions of the central nervous system (CNS). This review reports the CT and MRI findings of patients suffering from mucopolysaccharidosis in the context with aspects of biochemistry, pathophysiology, histological findings and clinical course. Neuroradiological findings include hydrocephalus, focal or global brain atrophy, and spinal cord compression at the atlantoaxial joint. Cribriform white matter lesions, corresponding to mucopolysaccharide deposition in the enlarged perivascular space, combined with leukencephalopathy are found in about half of the patients. Another half of the patients show minor to severe myelopathy at the atlantoaxial joint due to cervical cord compression. These findings are observed mainly in patients with MPS type IV, but as well in nearly all patients with MPS type VI. In the literature brain involvement is described exclusively in MPS type I, II and III, but the authors show definite demyelination and widening of the perivascular space in a number of patients with MPS type VI, who additionally suffered from progressive visual deficits, caused by ocular and/or optic nerve involvement.

Mucopolysaccharidosis type VI (MPS VI; Maroteaux-Lamy syndrome) is a lysosomal storage disorder caused by mutations in the N-acetylgalactosamine-4-sulfatase (arylsulfatase B, ARSB) gene. ARSB is a lysosomal enzyme involved in the degradation of the glycosaminoglycans (GAG) dermatan and chondroitin sulfate. ARSB mutations reduce enzyme function and GAG degradation, causing lysosomal storage and urinary excretion of these partially degraded substrates. Disease onset and rate of progression is variable, producing a spectrum of clinical presentation. In this study, 105 MPS VI patients—representing about 10% of the world MPS VI population—were studied for molecular genetic and biochemical parameters. Direct sequencing of patient genomic DNA was used to identify ARSB mutations. In total, 83 different disease-causing mutations were found, 62 of which were previously unknown. The novel sequence changes included: 38 missense mutations, five nonsense mutations, 11 deletions, one insertion, seven splice-site mutations, and four polymorphisms. ARSB mutant protein and residual activity were determined on fibroblast extracts for each patient. The identification of many novel mutations unique to individuals/their families highlighted the genetic heterogeneity of the disorder and provided an appropriate cohort to study the MPS VI phenotypic spectrum. This mutation analysis has identified a clear correlation between genotype and urinary GAG that can be used to predict clinical outcome. Hum Mutat 28(9), 897–903, 2007. Published 2007 Wiley-Liss, Inc.

Background: Fabry disease (FD) is a lysosomal storage disorder that is associated with marked cerebrovascular disease. Conventional MRI shows an extensive load of white matter lesions (WMLs) already at an early stage in FD. Objective: Investigator independent and sensitive quantification of brain structural changes in clinically affected men and women with FD. Methods: The authors performed a voxel-based analysis of diffusion tensor images (DTI) in 25 FD patients and 20 age-matched normal controls. Results: DTI revealed significant increases of cerebral white matter diffusivity (MD) in FD patients that were pronounced in the periventricular white matter. Even the subgroup of patients without significant WMLs-load (n = 18) showed increased diffusivity in the cerebral white matter. In gray matter areas MD elevation could only be detected in the posterior part of the thalamus, independently from visible pulvinar alterations on the T1-weighted images. Voxel-based anisotropy (FA) measurements did not differ significantly between patients and controls. Conclusions: The present study demonstrates the clinical feasibility of voxel-based analysis of DTI as a sensitive tool to quantify brain tissue alterations in FD. The pattern of increased brain tissue diffusivity is probably due to microangiopathic alterations mainly affecting the long perforating arteries.

OBJECTIVE. Our objective was to evaluate the safety, pharmacokinetics, and efficacy of laronidase in young, severely affected children with mucopolysaccharidosis I. METHODS. This was a prospective, open-label, multinational study of 20 patients who had mucopolysaccharidosis I and were RESULTS. Laronidase was well tolerated at both dosages. Investigators reported improved clinical status in 94% of patients at week 52. The mean urinary glycosaminoglycan level declined by ∼50% at week 13 and was sustained thereafter. A more robust decrease in urinary glycosaminoglycan was observed in patients with low antibody levels and those who were receiving the 200 U/kg dosage. On examination, the liver edge was reduced by 69.5% in patients with a palpable liver at baseline and week 52 (n = 10). The proportion of patients with left ventricular hypertrophy decreased from 53% to 17%. Global assessment of sleep studies showed improvement or stabilization in 67% of patients, and the apnea/hypopnea index decreased by 5.8 events per hour (−8.5%) in those with abnormal baseline values. The younger patients with Hurler syndrome ( CONCLUSIONS. Laronidase seems to be well tolerated and to provide clinical benefit in patients who have severe mucopolysaccharidosis I and are

Background: Fabry disease is a multisystemic life-threatening lysosomal storage disorder caused by deficiency of α-galactosidase A. Symptoms of the disease may occur in different organs including kidney, heart, and the nervous system. Objectives: To evaluate the nature and prevalence of pain in a large cohort of patients with Fabry disease and to assess the effect of enzyme replacement therapy (ERT) with agalsidase alfa. Methods: Retrospective analysis of the data of 752 patients with Fabry disease (393 females, 353 males) enrolled in the Fabry Outcome Survey, a multicentre database. Results: The prevalence of pain in male patients was 81.4% (females 65.3%). Mean age at onset of pain was 14.8 ±1.0 year in males (females 19.8 ± 1.4y). Pain was most frequently reported in the hands (males 76%, females 60%) and feet (males 73%, females 52%), but often affected the whole body. Interference of pain with daily life was higher in females than in males, and was observed predominantly for general activities, mood, and normal work. Fifty-eight percent of the patients were on ERT with agalsidase alfa. At 24 and 36 months after commencement of ERT, pain severity classification shifted towards lower severity (P < 0.05). Moreover, after 36 months, "average pain" and "pain now" were significantly reduced (P < 0.05). Conclusions: Pain is one of the most prevalent symptoms in Fabry disease with onset early in childhood. ERT with agalsidase alfa significantly reduces pain in this debilitating disorder.

Peridinin-chlorophyll a protein (PCP) is a unique water soluble antenna complex that employs the carotenoid peridinin as the main light-harvesting pigment. In the present study the near edge X-ray absorption fine structure (NEXAFS) spectrum of PCP was recorded at the carbon K-edge. Additionally, the NEXAFS spectra of the constituent pigments, chlorophyll a and peridinin, were measured. The energies of the lowest unoccupied molecular levels of these pigments appearing in the carbon NEXAFS spectrum were resolved. Individual contributions of the pigments and the protein to the measured NEXAFS spectrum of PCP were determined using a "building block" approach combining NEXAFS spectra of the pigments and the amino acids constituting the PCP apoprotein. The results suggest that absorption changes of the pigments in the carbon near K-edge region can be resolved following excitation using a suitable visible pump laser pulse. Consequently, it may be possible to study excitation energy transfer processes involving "optically dark" states of carotenoids in pigment-protein complexes by soft X-ray probe optical pump double resonance spectroscopy (XODR).

Aim: Left ventricular (LV) hypertrophy is a common feature in Fabry disease-related progressive infiltrative hypertrophic cardiomyopathy and affects both men and women, but at different ages. To date, however, little is known about the role of right ventricular (RV) function in Fabry disease. Therefore, this study aimed to investigate the extent of RV involvement in patients with Fabry disease. Methods: Echocardiographic examination of the right and left ventricle was carried out in 129 patients (80 women and 49 men) with Fabry disease. Results: RV hypertrophy was present in 46 patients (35.7%). Of these patients, 13 showed signs of severely depressed right systolic function (tricuspid annulus movement

Background To explore long-term effects of agalsidase alfa on Fabry disease cardiomyopathy in adults. Methods Retrospective analysis of prospectively collected data at a single center in Mainz, Germany, revealed that 45 adult patients (21 men, 24 women) had received agalsidase alfa for approximately 10 years. Data were extracted for cardiac and heart failure status, echocardiographic evaluations of cardiac structure and function, and renal function at treatment start and during agalsidase alfa treatment. Results After 10 years of agalsidase alfa treatment, heart failure classification had improved by at least 1 class in 22/42 patients, and angina scores were stable or improved in 41/42 patients. During treatment, no patients without left ventricular hypertrophy (LVH) at treatment initiation developed LVH, and no patients with LVH at treatment initiation showed a decline in left ventricular mass. Conclusions Approximately 10 years of agalsidase alfa treatment appeared to have beneficial effects for controlling progression and improving some symptoms of Fabry-associated cardiomyopathy.

Diagnostic imaging of head and neck cancer has made enormous progress during recent years. Next to morphological imaging modalities (computed tomography [CT] and magnetic resonance imaging [MRI]), there are also hybrid imaging systems that combine functional and morphological information (positron emission tomography [PET]/CT and PET/MRI). The aim of this study was to compare the diagnostic accuracy of PET/MRI in the diagnosis of head and neck cancer with other imaging modalities (MRI, CT, PET/CT). Ten patients (nine male and one female) with histologically proven oral squamous cell carcinoma participated in an 18 F-FDG-PET/CT scan and an additional 18 F-FDG PET/MRI scan prior to surgery. The morphological and functional results were compared with the histological results. Inclusion criteria were histologically proven oral squamous cell carcinoma and no prior surgical intervention, medical therapy, or local external radiation. There was no significant correlation between tumor differentiation and maximum standard uptake values. Functional imaging showed a slightly better correlation with the measurement of the maximal tumor diameter, whereas pure morphological imaging showed a better correlation with the measurement of infiltration depth. Only with PET/MRI could correct lymph node staging be reached; the other imaging tools showed false-negative or false-positive results. In conclusion, we showed in our limited patient cohort that PET/MRI is superior to the morphological imaging modalities, especially for lymph node staging.