High CK Research Articles

Overexpression of farnesyl pyrophosphate synthase increases myocardial ischemia/reperfusion injury in mice

Farnesyl pyrophosphate synthase (FPPS) is a vital enzyme in the mevalonate pathway. Our previous study has indicated that overexpression of FPPS increases hypoxia/reoxygenation (HR) injury in Heart-derived H9c2 Cells. Hence, we designed this experiment to further investigate the effect of FPPS on myocardial ischemia/reperfusion (MIR) injury using a transgenic (Tg) model, and explore the relevant mechanisms. The results showed that when mouse hearts were subjected to ex vivo I/R, Tg mice have a higher CK and LDH, a larger myocardial infarct size and lower heart function recovery. These phenomena are associated with the increased Rac1 activity and ROS generation. These findings point to that FPPS might be a potential target in preventing MIR in vivo.

P.337 - Multilevel molecular analysis identifies all dystrophin gene mutations pointing out that DMD is a genetically homogenous disease: repercussions on diagnosis, prevention and therapy

Dystrophinopathies are a group of allelic diseases caused by mutation in the dystrophin (DMD) gene: Deletions (~65%), duplications (~10%), small mutations (25%) and deep intronic mutations and complex rearrangements (~1%). Many exceptions do exist to the reading frame rule and a fine genetic characterization is mandatory for providing genetic diagnosis and making patients eligible for novel personalized trials. We analyzed the dystrophin gene in a large heterogeneous cohort of 891 individuals; reasons for testing were: clinical diagnosis of DMD (485), clinical diagnosis of BMD (226), symptomatic female carriers (43), asymptomatic patient with high CK (38), limb girdle muscle dystrophy (24) and at risk females tested for known familial mutation (78). We identified the dystrophin causative mutation in all the probands except for two boys with a clinical diagnosis of DMD and absence of dystrophin in the biopsy; in both the transcript analysis pinpointed the presence of a dystrophin gene mutation. Our data demonstrate that dystrophinopathies are genetically homogenous and a multilevel diagnostic approach is able to identify all DMD mutations; therefore a monogenic NGS approach should be endorsed in this disease. These findings carry some consequences in the address and interpretation of dystrophin genetic tests in clinical genetic counseling, in the definition of the role of modifiers, in the design of new therapies and in the plan of clinical care flowcharts.

P.359 - Different kinetic profiles of 2 patients presenting with muscle b-enolase deficiency

We describe the kinetic profile of total enolase activity in muscle tissue of two patients presenting with rhabdomyolysis due to muscle b-enolase deficiency. Two patients presenting with recurrent rhabdomyolysis were diagnosed by next generation sequencing with compound heterozygous mutations in ENO3. Baseline CK was normal in both patients. Rhabdomyolysis was confirmed by high serum CK levels (CK 75,000 IU/L and 193,000 IU/L) during acute episodes of muscle pain following exercise. Muscle biopsy showed minimal changes. Other signs and symptoms included exercise related myalgia and low lactate and ammonia production on a non-ischaemic forearm exercise test. Biochemical studies showed differing kinetic profiles in enolase activity. Muscle b-enolase deficiency is a rare cause of rhabdomyolysis. Here we report two cases with different kinetic profiles of total enolase activity in muscle tissue, illustrating that the disease is not always caused by null mutations, but instead can be caused by mutations that lower the maximal work rate of the enzyme (V max).

P.142 - Novel recessive splice site mutation in POPDC1 (BVES) is associated with first-degree atrioventricular block and muscular dystrophy

The Myocapture project using exome sequencing was set up to identify novel genes involved in neuromuscular disorders. The Institute of Myology's cohort of 98 families was selected as partner of this French project, after extensive characterisation of the clinical phenotype, muscle imaging and histopathology. Within the cohort, one consanguineous family presented two affected siblings. The index case, was first assessed at age 19 after a heart attack. He showed a first-degree atrioventricular (AV)-block associated with a CPK level as high as 4400 IU/L. Despite having a myopathic pattern at EMG and the dystrophic appearance of the muscle biopsy done two years later, no muscle weakness was reported. His elder sister had high CK levels (>1500 U/l) with a first-degree AV-block. However, she complained of muscle pain by the age of 30. Three other siblings were unaffected. Exome sequencing of the mother and the two affected children was performed. Variants in 8 candidate genes, including a homozygous one in POPDC1 (BVES), were identified after applying a filter of variant's frequency below 1% and a recessive mode of inheritance. POPDC1 (BVES) is a member of the Popeye-domain-containing (POPDC) gene family, which encodes transmembrane cAMP-binding proteins present in skeletal muscle and heart. Null mutations in animal models are associated with severe cardiac arrhythmia and muscular dystrophy. A single family with limb-girdle muscular dystrophy and AV-block has been described carrying a recessive missense mutation in POPDC1 (BVES). Here we describe a new homozygous variant. The nucleotide change is found in position +2 of intron 8 (c.816 + 2T>C). RT-PCR analyses of muscle RNA revealed exon 8 skipping and decreased expression of POPDC1 (BVES). Interestingly, plasma membrane localisation of POPDC1 and -2 protein was significantly reduced in the patient's muscle biopsies. These data are highly suggestive of a deleterious effect of this variant in this family.

Rhabdomyolysis in an HIV cohort: epidemiology, causes and outcomes

BackgroundThe Literature on rhabdomyolysis in the HIV-positive population is sparse and limited. We aimed to explore the incidence, patient characteristics, etiologies and outcomes of rhabdomyolysis in a cohort of HIV-positive patients identified through the Johns Hopkins HIV clinical registry between June 1992 and April 2014.MethodsA retrospective analysis of 362 HIV-positive patients with non-cardiac CK elevation ≥1000 IU/L was performed. Both inpatients and outpatients were included. Incidence rate and potential etiologies for rhabdomyolysis were ascertained. The development of acute kidney injury (AKI, defined as doubling of serum creatinine), need for dialysis, and death in the setting of rhabdomyolysis were determined. Logistic regression was used to evaluate the association of peak CK level with the development of AKI.ResultsThree hundred sixty two cases of rhabdomyolysis were identified in a cohort of 7079 patients with a 38,382 person years follow-up time. The incidence rate was nine cases per 1000 person-years (95% CI: 8.5–10.5). Infection was the most common etiology followed by compression injury and drug/alcohol use. One-third of cases had multiple potential etiologies. AKI developed in 46% of cases; 20% of which required dialysis. Thirteen percent died during follow-up. After adjustment, AKI was associated with higher CK (OR 2.05 for each 1-log increase in CK [95% CI: 1.40–2.99]), infection (OR 5.48 [95% CI 2.65–11.31]) and higher HIV viral load (OR 1.22 per 1-log increase [95% CI: 1.03–1.45]).ConclusionRhabdomyolysis in the HIV-positive population has many possible causes and is frequently multifactorial. HIV-positive individuals with rhabdomyolysis have a high risk of AKI and mortality.

P.074 Myopathic aspects of Mowat-Wilson Syndrome

Background: Mowat-Wilson Syndrome (MWS) is a genetic syndrome (ZEB2, OMIM: 235730) that occurs in 1 in 50000 births. It is characterized by microcephaly, intellectual disability, dysmorphisms (prominent chin, cupped ears, broad nasal bridge) and Hirschsprung’s disease. Although motor delay and hypotonia are common components, a myopathy has not been described in MWS literature. A childhood case with myopathic features prompted further study of this rare disease. Methods: Patients were recruited from the Mowat-Wilson Foundation via email or social media to complete a survey. Results: Thirteen surveys were returned to date. Although 54% of the patients reported motor delay, none of the patients had myopathy investigations. The index patient, presented at 1 year old, with hypotonia and developmental delay. Pregnancy and family history were unremarkable. Investigations revealed high CK levels (range 300 to 500 U/L), EMG confirmed myopathic motor units, and muscle biopsy showed type 1 fibre predominance. Single gene sequencing revealed pathogenic mutations of ZEB2, confirming a diagnosis of MWS. Conclusions: The description of myopathic features expands the spectrum of this rare syndrome and adds to the differential diagnosis of hyperCKemia in early childhood.

Risk Stratification of Untreated Mantle Cell Lymphoma Patients Using MIPI, Ki67 Proliferative Index and Cytogenetics

Background:There are multiple prognostic markers used to risk stratify pts with mantle cell lymphoma (MCL) including the MCL International Prognostic Index (MIPI) which divides newly diagnosed patients into three groups based on available clinical and laboratory data.Complex karyotype (CK) defined as equal to or greater than 3 chromosomal abnormalities has also been associated with inferior outcomes in MCL although its association with MIPI is lesswell-described. We conducted a multicenter analysis of MCL pts to assess the relative contributions of MIPI, CK and Ki67 as predictors of progression-free (PFS) and overall survival (OS).Methods:Untreated pts atleast18 years of age with MCL from 5 academic centers diagnosed between 1993 and 2015 who had available data describing MIPI, Ki67, and cytogenetics were included. Patients with 3 or more chromosomal abnormalities were defined as having CK. High Ki67 was defined as immunohistochemistry staining >30%. We described the associations between MIPI risk group, CK, Ki67, and other clinical factors of interest as predictors of PFS and OS in univariate and multivariable models.Results:Of 92 eligible MCL patients, 30 (32.6%) had low-risk, 36 pts (39.1%) had intermediate-risk, and 26 pts (28.3%) had high-risk MIPI scores. Ki67 index was >30% in 44.6% of patients, and 17.4% had CK. Neither Ki67 > 30% (p=0.548) norpresence of CK (p=0.409) wereassociated with MIPI risk category. In the low-risk MIPI subgroup, 37% of patients had Ki67 > 30% and 10% had CK. Induction regimens included: R-HyperCVAD (n=15), R-CHOP/R-DHAP (n=7), Nordic regimen (n=13), R-CHOP (n=10), B-R (n=19), and others (n=28). Thirty-two patients (39%) had an autologous stem cell transplant (ASCT) in first remission. Median PFS for the entire cohort was 3.8 years, and median OS has not yet been reached. Median PFS for was 2.2 years, 2.5 years, and 7.9 years for patients with high-, intermediate-, and low- risk MIPI scores, respectively, at diagnosis. Median PFS was 2.4 years for patients with Ki67 > 30% and 7.9 years for patients with Ki67 < 30%. Median PFS was 1.3 years for patients with CK and 4.3 years for patients without CK. High risk MIPI (p=0.042), CK (p=0.002), and Ki67 index > 30% (p=0.002) were all associated with inferior PFS in a univariable analyses (See Figure). In a multivariable analysis (See Table) Ki67 >30% (HR=3.66, p<0.001), and complex karyotype (HR=3.9, p=0.002) were associated with inferior PFS while autologous stem cell transplantation in first CR was associated with improved PFS (HR=0.21, p=0.002). MIPI risk score was not independently associated with PFS.Conclusions:Our findings suggest that Ki67> 30% and CK are independently associated with inferior outcomes in MCL. In this analysis, MIPI risk score was not independently associated with PFS or OS when considering these additional prognostic markers, which needs to be confirmed in other retrospective and prospective cohort studies. Our high-risk MIPI patients, in general, have an improved outcome compared to prior reports. (Hoster2008, 2014;Lex2014), suggesting that alternative markers may play an important role in MCL risk-assessment. Efforts to incorporate additional patients and develop a new prognostic model in MCL are underway. [Display omitted] [Display omitted] DisclosuresDanilov:Pharmacyclics: Consultancy; Astra Zeneca: Research Funding; Takeda: Research Funding; GIlead Sciences: Research Funding; ImmunoGen: Consultancy; Dava Oncology: Honoraria; Prime Oncology: Honoraria. Hamadani:Janssen: Consultancy; Celgene: Honoraria, Research Funding; Takeda Pharmaceuticals: Research Funding. Flowers:TG Therapeutics: Research Funding; Genentech: Consultancy, Research Funding; Infinity: Research Funding; Millenium/Takeda: Research Funding; Mayo Clinic: Research Funding; NIH: Research Funding; Roche: Consultancy, Research Funding; Pharmacyclics, LLC, an AbbVie Company: Research Funding; ECOG: Research Funding; AbbVie: Research Funding; Acerta: Research Funding; Gilead: Consultancy, Research Funding. Cohen:Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Infinity: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees; Pharmacyclics: Consultancy, Membership on an entity's Board of Directors or advisory committees; Millennium/Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Seattle Genetics: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Bristol-Myers Squibb: Research Funding.

Активность сывороточных ферментов в мониторинге тренировочного процесса высококвалифицированных спортсменов циклических видов спорта

The article presents the results of serum enzymes study in endurance sports. 311 elite athletes were investigated in swimming, biathlon, cycling, cross country skiing, rowing and kayaking. Capillary blood examination was performed on training camps during ten years. We included in statistical analysis 6950 CK tests, 5904 - AST and 5667- ALT. They were analyzed depending on kind of sport, training stage preparation and gender. The physiological serum enzyme ranges of elite athletes were estimated in view of sport, gender differences and stage of preparation. Serum enzyme activity trend was found in endurance sports at various preparation stages of the annual cycle of training. The high individual variation of CK activity was in endurance sports (variation coefficient 65,0-102,9% for men and 44,0-128,8% for women). It revealed the significantly higher CK activity in men compared to women. The trend of CK activity decrease was found from preparatory to competitive period. We examined the excess frequency enzyme activity higher upper limit of reference ranges for healthy individuals. It was 29,0% for CK, 23,2% - AST and 9,3% - ALT.

Proof of concept: hypovolemic hyponatremia may precede and augment creatine kinase elevations during an ultramarathon.

It is not known if exercise-associated hyponatremia (EAH) is a cause or consequence of exertional rhabdomyolysis (ER).We hypothesized that osmotic stress (EAH) coupled with mechanical stress (running) potentiated muscle cell breakdown (ER). This concept would be supported if a nadir in serum sodium concentration ([Na(+)]) temporally preceded peak creatine kinase levels (CK) during an ultramarathon run. Fifteen participants ran ≥104 km and had blood drawn: prior to start; 53; 104 km; and 24-h post run. Serum [Na(+)], CK, urea, creatinine and estimated glomerular filtration rate (eGFR) were measured from serial blood samples. Two-way repeated-measures ANOVA was used to examine differences regarding both race distance and natremia status. Ten of 15 participants demonstrated EAH (serum [Na(+)] <135 mmol/L) at least once during serial testing. Participants were categorized post hoc into one of three natremia groups based on lowest recorded [Na(+)]: (1) <129 mmol/L (n = 3; moderate EAH); (2) between 129 and 134 mmol/L (n = 7; mild EAH); and (3) >134 mmol/L (n = 5; normonatremia). Participants with lowest [Na(+)] demonstrated highest CK values at subsequent checkpoints. Significant natremia group differences noted at the 53 km point (p = 0.0002) for [Na(+)] while significant natremia group effect noted for CK seen at the 24-h post-finish testing point (p = 0.02). Significant natremia group effects noted for renal biomarkers, with the moderate EAH group documenting the lowest eGFR (p = 0.005), and highest serum urea (p = 0.0006) and creatinine (p < 0.0001) levels. Hyponatremic runners had lower post-race urine [Na(+)] than normonatremic runners (26 ± 15 vs. 89 ± 79 mmol/L; p = 0.03). Preliminary data support the possibility that transient hypovolemic EAH may precede and augment CK during an ultramarathon.

The effects of monovalent metal ions on the conformation of human telomere DNA using analytical ultracentrifugation.

A human telomere DNA segment (HT-DNA) can fold into a G-quadruplex in the presence of some monovalent cations. These cations can interact with the phosphate groups of the DNA segment and/or with the O6 oxygen atom of guanines, which are called non-specific interactions and specific interactions, respectively. However, until now how these two interactions affect the structure of HT-DNA has not been well understood. In this study, a combination of analytical ultracentrifugation (AUC) and circular dichroism (CD) was used to explore the effects of these two interactions on the structure of a 22-mer single-stranded DNA with a sequence of 5'-AGGG(TTAGGG)3-3'. The results showed that the standard sedimentation coefficient (s20,w) of HT-DNA starts to increase when the concentration of potassium ions (CK(+)) is higher than 10.0 µM due to the formation of a G-quadruplex through specific interactions. Whereas, for a control DNA, a higher CK(+) value of 1.0 mM was needed for increasing s20,w due to non-specific interactions. Moreover, potassium ions could promote the formation of the G-quadruplex much more easily than lithium, sodium and cesium ions, presumably due to its appropriate size in the dehydrated state and easier dehydration. The molar mass of DNA at different cation concentrations was nearly a constant and close to the theoretical value of the molar mass of monomeric HT-DNA, indicating that what we observed is the structural change of individual DNA chains.

DERMATIOMIOSITE COM DOENÇA INTERSTICIAL PULMONAR - UMA ASSOCIAÇÃO COM ANTI-MDA-5

A dermatomiosite (DM) é uma doença auto-imune com atingimento cutâneo, muscular e pulmonar. O MDA-5 (melanoma differentiation-associated protein 5) foi recentemente descrito como alvo de uma resposta serológica específica da DM que está associada a maior risco de doença pulmonar. Doente do sexo feminino, 54 anos, com fraqueza muscular proximal, mialgias, incapacidade funcional marcada e sinais cutâneos de dermatomiosite. Analiticamente apresentava elevação da aldolase e CK, e electromiografia e biópsia muscular compatíveis com dermatomiosite. A TAC pulmonar revelou padrão em vidro despolido. O estudo de auto-imunidade mostrou anti-MDA-5 positivo forte. Não foi detectada neoplasia. O tratamento com prednisolona 20mg, hidroxicloroquina 400mg e metotrexato 20mg/semana levou a melhoria apenas discreta do quadro, pelo que se introduziu imunoglobulina endovenosa (IgIV), com melhor resposta. Descreve-se o caso de uma doente com DM e doença intersticial pulmonar grave, com anti-MDA-5 positivo, chamando a atenção para esta associação até há pouco desconhecida.

P88 – 2855: Deficient alpha-dystroglycan presenting with rhabdomyolysis

Objective To describe a case of deficient alpha-dystroglycan revealed by acute rhabdomyolysis. Rhabdomyolysis frequently occurs in metabolic myopathies induced by exercise or triggered by intercurrent disease. Furthermore, rhabdomyolysis has been described in muscular dystrophies, particularly Duchenne and Becker dystrophies. It has only rarely been reported in limb girdle muscular dystrophies (LGMD), even though it may be the first and unique presenting feature of some type of LGMD. Case description and results A 12 year old boy complained about fatigue after summer holidays with more intensive exercise than usually. As laboratory investigations showed mild elevation of serum AST (264 IU/l, N Conclusion Limb girdle muscular dystrophy type 2I (LGMD2I) is one of the most common autosomal recessive limb girdle muscular dystrophies presenting in childhood. It may present with acute rhabdomyolysis as the first sign. It should be included in the diagnostic work-up of rhabdomyolysis and persistent high CK levels should prompt muscle biopsy and/or molecular diagnosis.

Clinical and Treatment Management Decisions in Two Asymptomatic Late-Onset Pompe Disease Siblings – Further Evidence of Scoliosis as a Clinical Sentinel Sign for Juvenile Pompe Disease

disease patient was identifi ed by chance through his dermatologist due to partial alopecia. Through high CK (1,310 IU/L) and muscle/hepatic biomarkers (AST, 162 IU/L; ALT, 189 IU/L) a muscle biopsy was performed revealing vacuolated muscle pathology with H-E and PAS stains. A compound heterozygote pathogenic GAA genotype was detected (-32-13 T>G/ c.2560C>T). A series of clinical and laboratory evaluations identifi ed several abnormal clinical signs, despite the absence of clinical symptoms. As the patient had a positive DBS, diminished GAA activity in leukocytes (0.56 nml/h/mg protein – RV: 1.0–5.9), high urinary GLc4 levels on HPLC, abnormal FVC drop during sitting–supine transition (>14%), and abnormalities on his tongue and edema of his thighs, seen on MRI, the decision was taken to initiate enzymatic replacement therapy (ERT) with 20 mg/kg rhGAA (Myozyme®), even though no subjective clinical symptoms were present. After 4 years on ERT treatment and motor/respiratory rehabilitation programs, all clinical parameters worsened, especially CK levels, muscle MRI with an overall muscle substitution for fat with muscle mass volume reduction; and a higher FVC drop during sitting–supine transiti on (>18%) were noted. Based on these clinical and laboClinical and Treatment Management Decisions in Two Asymptomatic Late-Onset Pompe Disease Siblings – Further Evidence of Scoliosis as a Clinical Sentinel Sign for Juvenile Pompe Disease

The Effects of Exercise on Serum Chemistry, Non-Esterified Fatty Acid, Insulin and Glucagon Dynamics during a 400 Meter Sprint in Racing Greyhounds

The effects of Greyhound racing on plasma insulin, glucagon and non-esterified fatty acid have not been extensively studied. We sought to examine the effects of exercise in 19 greyhounds trialed over 400 meters on serum biochemistry, osmolality, nonessential fatty acids, and the major glucoregulatory hormones insulin and glucagon, before and after racing. Due to the non-traditional dietary patterns of many racing Greyhounds, the diet was analyzed for macro and micro-nutrient assessment to determine adequacy of the diet. Many significant biochemical changes were observed and agreed with previous studies, except for low resting glucose, low post-exercise globulin and phosphorus concentrations, and a high CK both before and after exercise. Insulin concentrations increased 10 minutes post-exercise which might be a result of the rise in serum glucose observed. Non-esterified fatty acids decreased 10 minutes post-exercise, as a likely result of the insulin facilitated cellular uptake of fatty acids early in recovery. Glucagon concentrations did not change after exercise despite the rise in insulin. The diet analysis found some mineral deficiencies in sodium, selenium, iodine and copper; and contained a far higher protein and fat content, being atypical of diets that have been shown to improve the performance in racing Greyhounds.

Clinical and molecular genetic study of limb-girdle muscular dystrophy type 2A in a Chinese family

Objective To analyze the clinical, muscle pathological features and molecular mutations in the 2 Chinese Han siblings with limb-girdle muscular dystrophy(LGMD) and conclude the phenotype/genotype correlations. Methods Clinical and muscle pathological data were collected.Genomic DNA of the two siblings and their parents were extracted using standard procedures from the peripheral leukocytes.A custom of targeted gene panel including 61 neuromuscular genes were designed by using the Agilent Sureselect Target Enrichment Kit.Targeted next generation sequencing(NGS) was performed in the proband, and CAPN3 gene mutation was verified with Sanger sequencing in the two siblings and their parents.The dbSNP138 and http: //www.dmd.nl were searched to determine the disease-causing mutations. Results The proband slowly showed muscle weakness profoundly with pelvic muscles, developed difficulty in squatting and standing and climbing stairs.She had a tight Achilles tendon, high CK level(1 908-9 241 IU/L), wi-thout winging scapula and hypertrophy calf.The affected brother was only diagnosed hyper CKemia. By using the targeted NGS, the two siblings possessed the same two compound heterozygous mutations(c.717delT and c. 2243G>A) in CAPN3 gene.The two mutations both were verified by Sanger sequencing and had been reported before. Conclusions LGMD is clinically and genetically heterogeneous, and targeted NGS is powerful in defining the causal mutation of LGMD and helpful in investigating the exact genotype/phenotype analysis. Key words: Limb-girdle muscular dystrophy 2A; CAPN3 gene; Next generation sequencing; Mutation

G.P.157: Clinical and pathological features associated with mutations in MICU1

Mutations in the MICU1 gene have been shown to cause a condition with overlapping clinical and pathological features between muscular dystrophy, congenital myopathy and mitochondrial disease. The molecular defect results in mitochondrial dysfunction secondary to disruption of calcium uptake. We present the clinical/pathological features in a cohort of 18 patients. Patients presented between birth and 8years with a mild, relatively static, proximal myopathy associated with high Creatinine Kinase (2000–10,000iu/L), learning difficulties and frequent microcephaly. At follow up (5–28yrs), all remained ambulant but variable extrapyramidal symptoms had developed in the majority by the end of the 1st decade. Other features suggestive of mitochondrial dysfunction included peripheral neuropathy, icthyosis, stroke like episodes, episodic weakness, ataxia and cataracts. Cardiomyopathy was not seen. Serum and CSF lactate were normal where tested. Two had low free carnitine. Other metabolic investigations were normal. One had abnormal signal change in globus pallidus on MRI at 10yrs. All had abnormal muscle biopsies, with myopathic features; diffuse variation in fibre size, increased internal and central nuclei and clusters of regenerating fibres without pronounced fibrosis or fatty infiltration. Necrotic fibers were rare and fibre typing was preserved. Focal areas devoid of mitochondria (minicores) were observed in both type I and type II fibres. Respiratory chain enzyme activity, where tested, was normal. All had mutations in the MICU1 gene, 2 different homozygous splice site mutations (12 and 4 cases respectively) and a deletion in a kindred of 2 cases. Although the numbers are small, there appears to be some correlation between genotype and phenotype. Testing for MICU1 mutations should be considered in children with a static proximal myopathy associated with a high CK and CNS involvement when immunohistochemical studies do not identify a protein defect.

G.P.49: RYR1 mutations in adults with acute rhabdomyolysis episodes

Recurrent rhabdomyolysis episodes in adults are commonly caused by glycogenosis or fatty-acid oxidation disorders. However, in many cases the etiology remains unknown, and recently mutations in RYR1 appeared as a possible cause of exercise induced rhabdomyolysis. We studied 14 adults who presented acute rhabdomyolysis (CK above 10,000 UI/l) triggered by exercise or fever, in whom McArdle disease, fatty acid oxidation disorders, and LPIN1 mutations were excluded. All patients underwent muscle biopsy in order to screen RYR1 mutation on cDNA. Potentially pathogenic mutations in RYR1 gene were detected in 5 unrelated patients. Mutations were the following: p.Val4847Leu, p.G593R, p R3539H, Gly2434Arg, p.Gln3461Pro, p.A1352T, p.A933T and p.Ser1342Gly. Two patients presented each 2 and 3 heterozygous variants. Baseline CK levels were elevated in all cases but one (400–700 UI/l). None of them complained of myalgia or exercise intolerance. Rhabdomyolysis were triggered by weight-training in two patients, viral infection in two cases, and retinoic acid treatment in the last case. Higher recorded CK levels were between 28,000 and 94,000 UI/l, without renal failure. Three patients had a unique episode, whereas two other had several rhabdomyolysis episodes. Muscle biopsies were either normal, or showed moderate structure disorganization. Two siblings of a patient, the father of another patient, and the brother of a third patient, also presented mutations in the RYR1 gene and high baseline CK level. Per-anesthetic malignant hyperthermia was noticed in a grand-mother of one patient. Mutations in RYR1 gene should be systematically searched in patients with exercise or fever induced rhabdomyolysis episodes, after having excluded common metabolic disorders. This diagnostic implies major consequences for genetic counselling in the families, due to the potential risk of anesthetic malignant for the patients and their relatives.

Overall Survival in Small Cell Lung Cancer Detected with Epithelial, Mesenchymal and Stem Cell Biomarkers

ABSTRACT Aim: Markers of epithelial to mesenchymal transition (EMT) and markers associated with cancer stem cells (CSC) have been associated with increased chemoresistance in small cell lung cancer (SCLC). We and others previously demonstrated that elevated circulating tumour cells (CTC) are associated with worse survival. In the current exploratory study we hypothesized that markers of EMT and CSC in tumour biopsies obtained at diagnosis were associated with overall survival in patients with SCLC. Methods: Biopsies selected from small cell lung cancer pts study published previously by our group [ Ann Oncol 2012;23:2937-42]. Epithelial markers [Cytokeratin 8,18,19 (CK; ab41825, Abcam), EPCAM (VU1D9, cellsignaling), E-cadherin (24E10, cellsignaling)] and mesenchymal markers [Vimentin (V9, Santa Cruz) and c-MET (EP1454Y, Abcam)] were used as biomarkers for EMT. SOX2 (L1D6A2, cellsignaling) and CD44 (IM7, Biolegend) were used as CSC markers. Scoring by two independent observers having no information about clinical outcome. Immuno-Reactive Score (IRS) was made by multiplying percentage (0-6) and intensity (0-3). IRS score>=5 was high, =2 low Results: From 38 patients (mean age 65 yrs [49-84], 18 female pts, ECOG performance score 1 [0-3], extensive disease stage 26 pts, CTC at baseline 19 [0-14040]) adequate material was available to cut at least 10 tumour containing slides for immunohistochemical staining. Staining intensity high/low: CK 10/27; Vimentin 23/15; CD44 19/19; SOX-2 17/21; cMET 19/19; IRS high/low EPCAM 20/18, E-cadherin 14/24. Combining primary tumour low c-MET intensity with high CK intensity and high E-Cadherin IRS resulted in a significantly worse survival vs the c-METhigh/CKlow/E-cadherinlow group, OS 10 vs 24 month, p=0.040. A trend towards a positive correlation between CTC and primary tumor c-METlow was seen, p=0.062. Conclusions: SCLC with an epithelial phenotype (E-Cadhigh, CKhigh, cMETlow) may give rise to more epithelial CTCs in circulation than those with an mesenchymal phenotype and have a worse prognosis. Disclosure: All authors have declared no conflicts of interest.

Explicit finite deformation analysis of isogeometric membranes

NURBS-based isogeometric analysis was first extended to thin shell/membrane structures which allows for finite membrane stretching as well as large deflection and bending strain. The assumed non-linear kinematics employs the Kirchhoff–Love shell theory to describe the mechanical behaviour of thin to ultra-thin structures. The displacement fields are interpolated from the displacements of control points only, and no rotational degrees of freedom are used at control points. Due to the high order Ck (k≥1) continuity of NURBS shape functions the Kirchhoff–Love theory can be seamlessly implemented. An explicit time integration scheme is used to compute the transient response of membrane structures to time-domain excitations, and a dynamic relaxation method is employed to obtain steady-state solutions. The versatility and good performance of the present formulation are demonstrated with the aid of a number of test cases, including a square membrane strip under static pressure, the inflation of a spherical shell under internal pressure, the inflation of a square airbag and the inflation of a rubber balloon. The mechanical contribution of the bending stiffness is also evaluated.

Serum cytokeratin 19 fragment in advanced lung cancer: could we eventually have a serum tumor marker?

Introduction:Lung cancer is one of the most lethal malignancies; however, no serum marker has been routinely recommended until now.Methods:This is a prospective case control study including two groups of patients: Group I—patients with advanced lung cancer and Group II—patients with benign lung disease as control. Serum cytokeratin 19 (CK19) fragment levels were measured at baseline by real-time polymerase chain reaction before first-line chemotherapy. The CK19 cut-off taken was 15-cycle threshold. The primary end point was the comparison of high CK19 in cases and controls. The secondary end point was the correlation between high CK19 and progressive disease (PD), progression-free survival, and overall survival (OS) in advanced lung cancer patients.Results:A total of 30 patients with advanced lung cancer (16 non-small and 14 small cell lung cancer) and 15 patients with benign lung disease were included and followed up during the period from October 2008 to October 2011 with median follow-up of one and half years. High CK19 was found in 90% of lung cancer cases as compared with 7% in controls (p < 0.001). High CK19 was found in all cases showing PD (p = 0.04). One-year OS in high CK was 61% as compared with 33% in normal CK (p = 0.1).Conclusion:Serum CK19 fragment is a potential diagnostic and prognostic marker for advanced lung cancer.