Identifikation und Charakterisierung zellulärer Zielproteine zur antiviralen Therapie der SARS-Coronavirus Infektion

The severe acute respiratory syndrome (SARS) was first observed in the Chinese province Guangdong in November 2002. The disease quickly spread around the globe via air travelling and caused a worldwide epidemic. Several research institutions together with the World Health Organisation (WHO) identified the SARS-coronavirus (SARS-CoV) as the causative agent of this disease. During the epidemic, about 8,000 people were infected with a mortality of approximately 10%. Although no new infections have been observed since the summer of 2003, a recurrence of the pathogen cannot be excluded. Up to now, no specific therapy against the virus have been available. Viruses contain a very compact genome, which does not encode all proteins necessary for independant replication. Thus, viruses necessarily depend on host proteins and have to interact directly with them. The analysis of protein-protein interactions between SARS-CoV and human host cells contributes to a better understanding of the viral replication and pathogenicity. Prior to this work, an automated, genome-wide yeast-two-hybrid (Y2H) screen between all 28 proteins of SARS-CoV and the gene products of three human cDNA libraries had been performed, and approximately 460, mostly new protein-protein interactions had been identified. The aim of this work was to confirm newly identified virus-host SARS-CoV protein interactions and to functionally analyse them to identify new targets for antiviral therapy. 89 newly identified protein-protein interactions were examined via a modified LUMIER binding-assay to confirm individual interactions. 37 out of 89 protein interactions were found to be positive, resulting in a confirmation rate of 42%. In subsequent functional analyses of protein-protein interactions between the SARS-CoV non-structural protein 1 (Nsp1) and proteins of the immunophilin family, two different functional consequences were observed. First, it could be shown that SARS-CoV Nsp1 boosts the expression of genes regulated via the calcineurin/NFAT-signalling cascade. The increased expression of NFAT-regulated genes in SARS-CoV infection may cause the cytokine dysregulation described in SARS patients which leads to severe lung tissue destructions and which correlates with high mortality. The considerably less harmful human coronavirus HCoV-NL63 and mouse coronavirus (MHV) did not boost the expression of NFAT-regulated genes. It was thus hypothesized that the therapy of the cytokine dysregulation with the immunosuppressive drug Cyclosporine A (CspA) might improve the course of the disease. In addition, it could be shown for the first time that the replication of the SARS-CoV can be inhibited by the immunosuppressive drug CspA. Subsequent experiments showed a similar inhibition of the viral replication of the less harmful human coronavirus HCoV-NL63 and HCoV-229E mediated by CspA. In cooperation with several groups of the ”SARS-Zoonose- Verbund”, further inhibition experiments were performed with animal coronaviruses like FCoV, IBV Bd and TGEV PUR46, which showed a similar antiviral effect of CspA. The two cellular proteins Cyclophilin A and FK506 binding-protein 1A were shown to be essential for viral replication of HCoV-NL63. The findings of this work may contribute to a better understanding of the interactions between SARS-CoV and infected host cells and their innate immune response. The application of the general coronaviral inhibitor CspA identified in this study and of non-immunosuppressive CspA analogues like DEBIO-025 procures promising options for anti-coronaviral therapy.Im November 2002 brach das Schwere Akute Atemwegssyndrom (severe acute respiratory syndrom, SARS) zum ersten Mal in der chinesischen Provinz Guangdong aus. Dieser Erreger verursachte aufgrund des internationalen Flugverkehrs erstmals eine weltweite Epidemie. Verschiedene Forschungseinrichtungen konnten in Zusammenarbeit mit der Weltgesundheitsorganisation (WHO) das SARS-assoziierte Coronavirus (SARS-CoV) als Erreger der schweren Krankheit identifizieren. Insgesamt wurden während der Epidemie etwa 8000 Menschen infiziert, von denen ca. 10 % verstarben. Obwohl seit Sommer 2003 keine Neuinfektionen mehr beobachtet wurden, kann ein erneutes Auftreten dieses Pathogens nicht ausgeschlossen werden. Bis heute steht keine spezifische Therapie gegen SARS-CoV zur Verfügung. Viren haben ein sehr kompaktes Genom, in dem nicht alle notwendigen Proteine kodiert sind, die für einen kompletten Infektionszyklus benötigt werden. Aus diesem Grund sind Viren ausnahmslos abhängig von den Protein-Protein-Interaktionen mit einer lebenden Wirtszelle. Die Analyse von Protein-Protein-Interaktionen zwischen dem SARS-CoV und der humanen Wirtszelle trägt zum besseren Verständnis der viralen Replikation und Pathogenität bei. Im Vorfeld dieser Arbeit wurde ein automatisierter, genomweiter Hefe-Zwei-Hybrid (H2H)-Screen zwischen allen 28 Proteinen des SARS-CoV und den Genprodukten von drei humanen cDNA-Banken durchgeführt, wobei ca. 460, zumeist völlig neue, Protein-Protein-Interaktionen zwischen dem SARS-CoV und dem humanen Wirt identifiziert wurden. Ziel dieser Arbeit war es, die neu identifzierten Protein-Protein-Interaktionen zu bestätigen und funktionelle Analysen ausgewählter Interaktionen durchzuführen, um neue Angriffspunkte für die antivirale Therapie zu finden. 89 Protein-Protein-Interaktionen, die im H2H-Screen neu identifiziert werden konnten, wurden mit Hilfe des modifizierten LUMIER Bindungs-Assays zur Bestätigung der einzelnen Interaktionen untersucht. Von diesen 89 getesteten Protein-Protein-Interaktionen waren 37 Tests positiv, wodurch sich eine Bestätigungsrate von 42 % ergab. In anschließenden funktionellen Analysen der Protein-Interaktionen zwischen dem SARS-CoV Nicht-Strukturprotein 1 (Nsp1) und Proteinen der Immunophilinfamilie konnten zwei Funktionen dieser Interaktionen aufgezeigt werden. Zunächst konnte gezeigt werden, dass das SARS-CoV Nsp1 die Expression von Genen, welche über die Calcineurin/NFAT-Signalkaskade reguliert werden, erhöht. Die SARS-spezifische Erhöhung der Expression NFAT-regulierter Gene kann eine Ursache der in SARS-Patienten beschriebenen Zytokindysregulation sein. Diese Zytokindysregulation führt zu schweren Gewebeschädigungen in der Lunge und trägt maßgeblich zum schlechten Ausgang der Krankheit bei. Das weniger pathogene humane Coronavirus HCoV-NL63 und das Maus-Coronavirus MHV zeigten diese Erhöhung der Expression NFAT-regulierter Gene nicht auf. Es wurde deshalb die Hypothese aufgestellt, dass eine Behandlung der Zytokindysregulation mit dem Immunsuppressivum Cyclosporin A positive Auswirkungen auf den Verlauf der Krankheit hat. Zum zweiten konnte erstmals gezeigt werden, dass die Replikation des SARS-CoV durch das Immunsuppressivum Cyclosporin A gehemmt werden kann. Anschließende Inhibitionsversuche der deutlich harmloseren humanen Coronaviren HCoV-NL63 und HCoV-229E zeigten die gleiche Hemmung der viralen Replikation durch Cyclosporin A. In Kooperation mit verschiedenen Arbeitsgruppen des SARS-Zoonose-Verbunds konnten weitere Inhibitionsversuche mit den Tiercoronaviren FCoV, IBV Bd und TGEV PUR46 durchgeführt werden und ebenfalls ein inhibitorisches Potential des Cyclosporin A auf die virale Replikation dieser Tiercoronaviren gezeigt werden. In weiterführenden Untersuchungen zum Wirkmechanismus der CspA- und FK506-vermittelten Inhibition der Replikation des humanen Coronavirus HCoV-NL63 konnten die beiden zellulären Proteine Cyclophilin A und FK506-Bindeprotein1A (FKBP1A) erstmals als essentielle Proteine für die virale Replikation identifiziert werden. Die Erkenntnisse dieser Arbeit können dazu beitragen, die komplexen Interaktionen zwischen dem SARS-CoV, der infizierten Wirtszelle und der Immunabwehr besser zu verstehen. Weiterhin konnte im Rahmen dieser Arbeit ein allgemeiner, coronaviraler Inhibitor in Form von Cyclosporin A identifiziert werden. Nicht-immunsuppressive Cyclosporin A Analoga wie DEBIO 025 sind deshalb mögliche Kandidaten für die Therapie coronaviraler Infektionen

Identifikation und Charakterisierung zellulärer Zielproteine zur antiviralen Therapie der SARS-Coronavirus Infektion

The severe acute respiratory syndrome (SARS) was first observed in the Chinese province Guangdong in November 2002. The disease quickly spread around the globe via air travelling and caused a worldwide epidemic. Several research institutions together with the World Health Organisation (WHO) identified the SARS-coronavirus (SARS-CoV) as the causative agent of this disease. During the epidemic, about 8,000 people were infected with a mortality of approximately 10%. Although no new infections have been observed since the summer of 2003, a recurrence of the pathogen cannot be excluded. Up to now, no specific therapy against the virus have been available. Viruses contain a very compact genome, which does not encode all proteins necessary for independant replication. Thus, viruses necessarily depend on host proteins and have to interact directly with them. The analysis of protein-protein interactions between SARS-CoV and human host cells contributes to a better understanding of the viral replication and pathogenicity. Prior to this work, an automated, genome-wide yeast-two-hybrid (Y2H) screen between all 28 proteins of SARS-CoV and the gene products of three human cDNA libraries had been performed, and approximately 460, mostly new protein-protein interactions had been identified. The aim of this work was to confirm newly identified virus-host SARS-CoV protein interactions and to functionally analyse them to identify new targets for antiviral therapy. 89 newly identified protein-protein interactions were examined via a modified LUMIER binding-assay to confirm individual interactions. 37 out of 89 protein interactions were found to be positive, resulting in a confirmation rate of 42%. In subsequent functional analyses of protein-protein interactions between the SARS-CoV non-structural protein 1 (Nsp1) and proteins of the immunophilin family, two different functional consequences were observed. First, it could be shown that SARS-CoV Nsp1 boosts the expression of genes regulated via the calcineurin/NFAT-signalling cascade. The increased expression of NFAT-regulated genes in SARS-CoV infection may cause the cytokine dysregulation described in SARS patients which leads to severe lung tissue destructions and which correlates with high mortality. The considerably less harmful human coronavirus HCoV-NL63 and mouse coronavirus (MHV) did not boost the expression of NFAT-regulated genes. It was thus hypothesized that the therapy of the cytokine dysregulation with the immunosuppressive drug Cyclosporine A (CspA) might improve the course of the disease. In addition, it could be shown for the first time that the replication of the SARS-CoV can be inhibited by the immunosuppressive drug CspA. Subsequent experiments showed a similar inhibition of the viral replication of the less harmful human coronavirus HCoV-NL63 and HCoV-229E mediated by CspA. In cooperation with several groups of the ”SARS-Zoonose- Verbund”, further inhibition experiments were performed with animal coronaviruses like FCoV, IBV Bd and TGEV PUR46, which showed a similar antiviral effect of CspA. The two cellular proteins Cyclophilin A and FK506 binding-protein 1A were shown to be essential for viral replication of HCoV-NL63. The findings of this work may contribute to a better understanding of the interactions between SARS-CoV and infected host cells and their innate immune response. The application of the general coronaviral inhibitor CspA identified in this study and of non-immunosuppressive CspA analogues like DEBIO-025 procures promising options for anti-coronaviral therapy.Im November 2002 brach das Schwere Akute Atemwegssyndrom (severe acute respiratory syndrom, SARS) zum ersten Mal in der chinesischen Provinz Guangdong aus. Dieser Erreger verursachte aufgrund des internationalen Flugverkehrs erstmals eine weltweite Epidemie. Verschiedene Forschungseinrichtungen konnten in Zusammenarbeit mit der Weltgesundheitsorganisation (WHO) das SARS-assoziierte Coronavirus (SARS-CoV) als Erreger der schweren Krankheit identifizieren. Insgesamt wurden während der Epidemie etwa 8000 Menschen infiziert, von denen ca. 10 % verstarben. Obwohl seit Sommer 2003 keine Neuinfektionen mehr beobachtet wurden, kann ein erneutes Auftreten dieses Pathogens nicht ausgeschlossen werden. Bis heute steht keine spezifische Therapie gegen SARS-CoV zur Verfügung. Viren haben ein sehr kompaktes Genom, in dem nicht alle notwendigen Proteine kodiert sind, die für einen kompletten Infektionszyklus benötigt werden. Aus diesem Grund sind Viren ausnahmslos abhängig von den Protein-Protein-Interaktionen mit einer lebenden Wirtszelle. Die Analyse von Protein-Protein-Interaktionen zwischen dem SARS-CoV und der humanen Wirtszelle trägt zum besseren Verständnis der viralen Replikation und Pathogenität bei. Im Vorfeld dieser Arbeit wurde ein automatisierter, genomweiter Hefe-Zwei-Hybrid (H2H)-Screen zwischen allen 28 Proteinen des SARS-CoV und den Genprodukten von drei humanen cDNA-Banken durchgeführt, wobei ca. 460, zumeist völlig neue, Protein-Protein-Interaktionen zwischen dem SARS-CoV und dem humanen Wirt identifiziert wurden. Ziel dieser Arbeit war es, die neu identifzierten Protein-Protein-Interaktionen zu bestätigen und funktionelle Analysen ausgewählter Interaktionen durchzuführen, um neue Angriffspunkte für die antivirale Therapie zu finden. 89 Protein-Protein-Interaktionen, die im H2H-Screen neu identifiziert werden konnten, wurden mit Hilfe des modifizierten LUMIER Bindungs-Assays zur Bestätigung der einzelnen Interaktionen untersucht. Von diesen 89 getesteten Protein-Protein-Interaktionen waren 37 Tests positiv, wodurch sich eine Bestätigungsrate von 42 % ergab. In anschließenden funktionellen Analysen der Protein-Interaktionen zwischen dem SARS-CoV Nicht-Strukturprotein 1 (Nsp1) und Proteinen der Immunophilinfamilie konnten zwei Funktionen dieser Interaktionen aufgezeigt werden. Zunächst konnte gezeigt werden, dass das SARS-CoV Nsp1 die Expression von Genen, welche über die Calcineurin/NFAT-Signalkaskade reguliert werden, erhöht. Die SARS-spezifische Erhöhung der Expression NFAT-regulierter Gene kann eine Ursache der in SARS-Patienten beschriebenen Zytokindysregulation sein. Diese Zytokindysregulation führt zu schweren Gewebeschädigungen in der Lunge und trägt maßgeblich zum schlechten Ausgang der Krankheit bei. Das weniger pathogene humane Coronavirus HCoV-NL63 und das Maus-Coronavirus MHV zeigten diese Erhöhung der Expression NFAT-regulierter Gene nicht auf. Es wurde deshalb die Hypothese aufgestellt, dass eine Behandlung der Zytokindysregulation mit dem Immunsuppressivum Cyclosporin A positive Auswirkungen auf den Verlauf der Krankheit hat. Zum zweiten konnte erstmals gezeigt werden, dass die Replikation des SARS-CoV durch das Immunsuppressivum Cyclosporin A gehemmt werden kann. Anschließende Inhibitionsversuche der deutlich harmloseren humanen Coronaviren HCoV-NL63 und HCoV-229E zeigten die gleiche Hemmung der viralen Replikation durch Cyclosporin A. In Kooperation mit verschiedenen Arbeitsgruppen des SARS-Zoonose-Verbunds konnten weitere Inhibitionsversuche mit den Tiercoronaviren FCoV, IBV Bd und TGEV PUR46 durchgeführt werden und ebenfalls ein inhibitorisches Potential des Cyclosporin A auf die virale Replikation dieser Tiercoronaviren gezeigt werden. In weiterführenden Untersuchungen zum Wirkmechanismus der CspA- und FK506-vermittelten Inhibition der Replikation des humanen Coronavirus HCoV-NL63 konnten die beiden zellulären Proteine Cyclophilin A und FK506-Bindeprotein1A (FKBP1A) erstmals als essentielle Proteine für die virale Replikation identifiziert werden. Die Erkenntnisse dieser Arbeit können dazu beitragen, die komplexen Interaktionen zwischen dem SARS-CoV, der infizierten Wirtszelle und der Immunabwehr besser zu verstehen. Weiterhin konnte im Rahmen dieser Arbeit ein allgemeiner, coronaviraler Inhibitor in Form von Cyclosporin A identifiziert werden. Nicht-immunsuppressive Cyclosporin A Analoga wie DEBIO 025 sind deshalb mögliche Kandidaten für die Therapie coronaviraler Infektionen

Fetal eye movements on magnetic resonance imaging.

OBJECTIVES: Eye movements are the physical expression of upper fetal brainstem function. Our aim was to identify and differentiate specific types of fetal eye movement patterns using dynamic MRI sequences. Their occurrence as well as the presence of conjugated eyeball motion and consistently parallel eyeball position was systematically analyzed. METHODS: Dynamic SSFP sequences were acquired in 72 singleton fetuses (17-40 GW, three age groups [17-23 GW, 24-32 GW, 33-40 GW]). Fetal eye movements were evaluated according to a modified classification originally published by Birnholz (1981): Type 0: no eye movements; Type I: single transient deviations; Type Ia: fast deviation, slower reposition; Type Ib: fast deviation, fast reposition; Type II: single prolonged eye movements; Type III: complex sequences; and Type IV: nystagmoid. RESULTS: In 95.8% of fetuses, the evaluation of eye movements was possible using MRI, with a mean acquisition time of 70 seconds. Due to head motion, 4.2% of the fetuses and 20.1% of all dynamic SSFP sequences were excluded. Eye movements were observed in 45 fetuses (65.2%). Significant differences between the age groups were found for Type I (p = 0.03), Type Ia (p = 0.031), and Type IV eye movements (p = 0.033). Consistently parallel bulbs were found in 27.3-45%. CONCLUSIONS: In human fetuses, different eye movement patterns can be identified and described by MRI in utero. In addition to the originally classified eye movement patterns, a novel subtype has been observed, which apparently characterizes an important step in fetal brainstem development. We evaluated, for the first time, eyeball position in fetuses. Ultimately, the assessment of fetal eye movements by MRI yields the potential to identify early signs of brainstem dysfunction, as encountered in brain malformations such as Chiari II or molar tooth malformations

MR-based morphometry of the posterior fossa in fetuses with neural tube defects of the spine.

OBJECTIVES: In cases of "spina bifida," a detailed prenatal imaging assessment of the exact morphology of neural tube defects (NTD) is often limited. Due to the diverse clinical prognosis and prenatal treatment options, imaging parameters that support the prenatal differentiation between open and closed neural tube defects (ONTDs and CNTDs) are required. This fetal MR study aims to evaluate the clivus-supraocciput angle (CSA) and the maximum transverse diameter of the posterior fossa (TDPF) as morphometric parameters to aid in the reliable diagnosis of either ONTDs or CNTDs. METHODS: The TDPF and the CSA of 238 fetuses (20-37 GW, mean: 28.36 GW) with a normal central nervous system, 44 with ONTDS, and 13 with CNTDs (18-37 GW, mean: 24.3 GW) were retrospectively measured using T2-weighted 1.5 Tesla MR -sequences. RESULTS: Normal fetuses showed a significant increase in the TDPF (r = .956; p<.001) and CSA (r = .714; p<.001) with gestational age. In ONTDs the CSA was significantly smaller (p<.001) than in normal controls and CNTDs, whereas in CNTDs the CSA was not significantly smaller than in controls (p = .160). In both ONTDs and in CNTDs the TDPF was significantly different from controls (p<.001). CONCLUSIONS: The skull base morphology in fetuses with ONTDs differs significantly from cases with CNTDs and normal controls. This is the first study to show that the CSA changes during gestation and that it is a reliable imaging biomarker to distinguish between ONTDs and CNTDs, independent of the morphology of the spinal defect

Temporal muscle thickness is an independent prognostic marker in melanoma patients with newly diagnosed brain metastases.

OBJECTIVES: The purpose of this study was to evaluate the prognostic relevance of temporal muscle thickness (TMT) in melanoma patients with newly diagnosed brain metastases. METHODS: TMT was retrospectively assessed in 146 melanoma patients with newly diagnosed brain metastases on cranial magnetic resonance images. Chart review was used to retrieve clinical parameters, including disease-specific graded prognostic assessment (DS-GPA) and survival times. RESULTS: Patients with a TMT > median showed a statistically significant increase in survival time (13 months) compared to patients with a TMT < median (5 months; p < 0.001; log rank test). A Cox regression model revealed that the risk of death was increased by 27.9% with every millimeter reduction in TMT. In the multivariate analysis, TMT (HR 0.724; 95% 0.642-0.816; < 0.001) and DS-GPA (HR 1.214; 95% CI 1.023-1.439; p = 0.026) showed a statistically significant correlation with overall survival. CONCLUSION: TMT is an independent predictor of survival in melanoma patients with brain metastases. This parameter may aid in patient selection for clinical trials or to the choice of different treatment options based on the determination of frail patient populations

The SARS-coronavirus-host interactome

Coronaviruses (CoVs) are important human and animal pathogens that induce fatal respiratory, gastrointestinal and neurological disease. The outbreak of the severe acute respiratory syndrome (SARS) in 2002/2003 has demonstrated human vulnerability to (Coronavirus) CoV epidemics. Neither vaccines nor therapeutics are available against human and animal CoVs. Knowledge of host cell proteins that take part in pivotal virus-host interactions could define broad-spectrum antiviral targets. In this study, we used a systems biology approach employing a genome-wide yeast-two hybrid interaction screen to identify immunopilins (PPIA, PPIB, PPIH, PPIG, FKBP1A, FKBP1B) as interaction partners of the CoV non-structural protein 1 (Nsp1). These molecules modulate the Calcineurin/NFAT pathway that plays an important role in immune cell activation. Overexpression of NSP1 and infection with live SARS-CoV strongly increased signalling through the Calcineurin/NFAT pathway and enhanced the induction of interleukin 2, compatible with late-stage immunopathogenicity and long-term cytokine dysregulation as observed in severe SARS cases. Conversely, inhibition of cyclophilins by cyclosporine A (CspA) blocked the replication of CoVs of all genera, including SARS-CoV, human CoV-229E and -NL-63, feline CoV, as well as avian infectious bronchitis virus. Non-immunosuppressive derivatives of CspA might serve as broad-range CoV inhibitors applicable against emerging CoVs as well as ubiquitous pathogens of humans and livestock

High correlation of temporal muscle thickness with lumbar skeletal muscle cross-sectional area in patients with brain metastases.

OBJECTIVES: This study aimed to assess the correlation of temporal muscle thickness (TMT), measured on routine cranial magnetic resonance (MR) images, with lumbar skeletal muscles obtained on computed tomography (CT) images in brain metastasis patients to establish a new parameter estimating skeletal muscle mass on brain MR images. METHODS: We retrospectively analyzed the cross-sectional area (CSA) of skeletal muscles at the level of the third lumbar vertebra on computed tomography scans and correlated these values with TMT on MR images of the brain in two independent cohorts of 93 lung cancer and 61 melanoma patients (overall: 154 patients) with brain metastases. RESULTS: Pearson correlation revealed a strong association between mean TMT and CSA in lung cancer and melanoma patients with brain metastases (0.733; p<0.001). The two study cohorts did not differ significantly in patient characteristics, including age (p = 0.661), weight (p = 0.787), and height (p = 0.123). However, TMT and CSA measures differed significantly between male and female patients in both lung cancer and melanoma patients with brain metastases (p<0.001). CONCLUSION: Our data indicate that TMT, measured on routine cranial MR images, is a useful surrogate parameter for the estimation of skeletal muscle mass in patients with brain metastases. Thus, TMT may be useful for prognostic assessment, treatment considerations, and stratification or a selection factor for clinical trials in patients with brain metastases. Further studies are needed to assess the association between TMT and clinical frailty parameters, and the usefulness of TMT in patients with primary brain tumors

Sarcopenia in Neurological Patients: Standard Values for Temporal Muscle Thickness and Muscle Strength Evaluation.

Temporal muscle thickness (TMT) was investigated as a novel surrogate marker on MRI examinations of the brain, to detect patients who may be at risk for sarcopenia. TMT was analyzed in a retrospective, normal collective cohort (n = 624), to establish standard reference values. These reference values were correlated with grip strength measurements and body mass index (BMI) in 422 healthy volunteers and validated in a prospective cohort (n = 130) of patients with various neurological disorders. Pearson correlation revealed a strong association between TMT and grip strength (retrospective cohort, ρ = 0.746; p < 0.001; prospective cohort, ρ = 0.649; p < 0.001). A low or no association was found between TMT and age (retrospective cohort, R2 correlation coefficient 0.20; p < 0.001; prospective cohort, ρ = -0.199; p = 0.023), or BMI (retrospective cohort, ρ = 0.116; p = 0.042; prospective cohort, ρ = 0.227; p = 0.009), respectively. Male patients with temporal wasting and unintended weight loss, respectively, showed significantly lower TMT values (p = 0.04 and p = 0.015, unpaired t-test). TMT showed a high correlation with muscle strength in healthy individuals and in patients with various neurological disorders. Therefore, TMT should be integrated into the diagnostic workup of neurological patients, to prevent, delay, or treat sarcopenia

Survival prediction using temporal muscle thickness measurements on cranial magnetic resonance images in patients with newly diagnosed brain metastases.

OBJECTIVES: To evaluate the prognostic relevance of temporal muscle thickness (TMT) in brain metastasis patients. METHODS: We retrospectively analysed TMT on magnetic resonance (MR) images at diagnosis of brain metastasis in two independent cohorts of 188 breast cancer (BC) and 247 non-small cell lung cancer (NSCLC) patients (overall: 435 patients). RESULTS: Survival analysis using a Cox regression model showed a reduced risk of death by 19% with every additional millimetre of baseline TMT in the BC cohort and by 24% in the NSCLC cohort. Multivariate analysis included TMT and diagnosis-specific graded prognostic assessment (DS-GPA) as covariates in the BC cohort (TMT: HR 0.791/CI [0.703-0.889]/p < 0.001; DS-GPA: HR 1.433/CI [1.160-1.771]/p = 0.001), and TMT, gender and DS-GPA in the NSCLC cohort (TMT: HR 0.710/CI [0.646-0.780]/p < 0.001; gender: HR 0.516/CI [0.387-0.687]/p < 0.001; DS-GPA: HR 1.205/CI [1.018-1.426]/p = 0.030). CONCLUSION: TMT is easily and reproducibly assessable on routine MR images and is an independent predictor of survival in patients with newly diagnosed brain metastasis from BC and NSCLC. TMT may help to better define frail patient populations and thus facilitate patient selection for therapeutic measures or clinical trials. Further prospective studies are needed to correlate TMT with other clinical frailty parameters of patients. KEY POINTS: • TMT has an independent prognostic relevance in brain metastasis patients. • It is an easily and reproducibly parameter assessable on routine cranial MRI. • This parameter may aid in patient selection and stratification in clinical trials. • TMT may serve as surrogate marker for sarcopenia

Fetal eye movements on magnetic resonance imaging.

OBJECTIVES: Eye movements are the physical expression of upper fetal brainstem function. Our aim was to identify and differentiate specific types of fetal eye movement patterns using dynamic MRI sequences. Their occurrence as well as the presence of conjugated eyeball motion and consistently parallel eyeball position was systematically analyzed. METHODS: Dynamic SSFP sequences were acquired in 72 singleton fetuses (17-40 GW, three age groups [17-23 GW, 24-32 GW, 33-40 GW]). Fetal eye movements were evaluated according to a modified classification originally published by Birnholz (1981): Type 0: no eye movements; Type I: single transient deviations; Type Ia: fast deviation, slower reposition; Type Ib: fast deviation, fast reposition; Type II: single prolonged eye movements; Type III: complex sequences; and Type IV: nystagmoid. RESULTS: In 95.8% of fetuses, the evaluation of eye movements was possible using MRI, with a mean acquisition time of 70 seconds. Due to head motion, 4.2% of the fetuses and 20.1% of all dynamic SSFP sequences were excluded. Eye movements were observed in 45 fetuses (65.2%). Significant differences between the age groups were found for Type I (p = 0.03), Type Ia (p = 0.031), and Type IV eye movements (p = 0.033). Consistently parallel bulbs were found in 27.3-45%. CONCLUSIONS: In human fetuses, different eye movement patterns can be identified and described by MRI in utero. In addition to the originally classified eye movement patterns, a novel subtype has been observed, which apparently characterizes an important step in fetal brainstem development. We evaluated, for the first time, eyeball position in fetuses. Ultimately, the assessment of fetal eye movements by MRI yields the potential to identify early signs of brainstem dysfunction, as encountered in brain malformations such as Chiari II or molar tooth malformations