Atypical phenotypes in titinopathies explained by second tit

RESEARCH ARTICLE

Atypical Phenotypes in Titinopathies

Explained by Second Titin Mutations

Anni Evil€a,MSc,1Anna Vihola,PhD,1Jaakko Sarparanta,MSc,1 Olayinka Raheem,PhD,2Johanna Palmio,MD,PhD,2Satu Sandell,MD,2,3 Bruno Eymard,MD,PhD,4Isabel Illa,MD,PhD,5Ricard Rojas-Garcia,MD,PhD,5

Karolina Hankiewicz,MD,5Luis Negr~a o,MD,PhD,6Tuija L€o pp€o nen,MD,PhD,7

Pekka Nokelainen,MD,PhD,7Mikko K€a rpp€a,MD,PhD,8Sini Penttil€a,MSc,2 Mark Screen,MSc,1Tiina Suominen,PhD,2Isabelle Richard,PhD,9

Peter Hackman,PhD,1and Bjarne Udd,MD,PhD1,2,10

Objective:Several patients with previously reported titin gene(TTN)mutations causing tibial muscular dystrophy (TMD)have more complex,severe,or unusual phenotypes.This study aimed to clarify the molecular cause of the var-iant phenotypes in8patients of7European families.

Methods:Clinical,histopathological,and muscle imaging data of patients and family members were reanalyzed.The titin protein was analyzed by Western blotting and TTN gene by reverse transcription polymerase chain reaction(RT-PCR)and Sanger sequencing.

Results:Western blotting showed more pronounced C-terminal titin abnormality than expected for heterozygous probands,suggesting the existence of additional TTN mutations.RT-PCR indicated unequal mRNA expression of the TTN alleles in biopsies of6patients,3with an limb-girdle muscular dystrophy type2J(LGMD2J)phenotype.Novel frameshift mutations were identified in5patients.A novel A-band titin mutation,c.92167C>T(p.P30723S),was found in1patient,and1Portuguese patient with a severe TMD phenotype proved to be homozygous for the previ-ously reported Iberian TMD mutation.

Interpretation:The unequal expression levels of TTN transcripts in5probands suggested severely reduced expres-sion of the frameshift mutated allele,probably through nonsense-mediated decay,explaining the more severe phe-notypes.The Iberian TMD mutation may cause a more severe TMD rather than LGMD2J when homozygous.The Finnish patient compound heterozygous for the FINmaj TMD mutation and the novel A-band titin missense mutation showed a phenotype completely different from previously described titinopathies.Our results further expand the complexity of muscular dystrophies caused by TTN mutations and suggest that the coexistence of second mutations may constitute a more common general mechanism explaining phenotype variability.

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T ibial muscular dystrophy(TMD;Online Mendelian Inheritance in Man database[OMIM]#600334)is an autosomal dominant distal myopathy caused by mutations in the titin gene(TTN).The disease is charac-terized by weakness and atrophy especially in the anterior compartment muscles of the lower leg,tibialis anterior,

View this article online at https://www.wendangku.net/doc/d34611052.html,.DOI:10.1002/ana.24102

Received Oct22,2013,and in revised form Dec23,2013.Accepted for publication Dec25,2013.

Address correspondence to Dr Hackman,Folkh€a lsan Institute of Genetics and Department of Medical Genetics,Haartman Institute,University of

Helsinki,Helsinki,Finland.E-mail address:peter.hackman@helsinki.fi

From the1Folkh€a lsan Institute of Genetics and Department of Medical Genetics,Haartman Institute,University of Helsinki,Helsinki,Finland;

2Neuromuscular Research Center,University of Tampere and Tampere University Hospital,Tampere,Finland;3Sein€a joki Central Hospital,Department of Neurology,Sein€a joki,Finland;4Institute of Myology,National Reference Center for Neuromuscular Disorders,University Hospital of Salp^e trie`re,Paris, France;5Unitat Patologia Neuromuscular,Servei Neurologia,Hospital Santa Creu i Sant Pau,Universitat Aut o noma,Barcelona,Spain;6Neuromuscular Unit,Neurology Department,Coimbra University Hospital,Coimbra,Portugal;7Department of Child Neurology,Kuopio University Hospital,Kuopio,Fin-land;8Department of Clinical Medicine,Neurology,University of Oulu and Clinical Research Center,Oulu University Hospital,Oulu,Finland;9G e n e thon, Evry,France;and10Neurology Department,Vaasa Central Hospital,Vaasa,Finland.

Additional supporting information can be found in the online version of this article.

230V C2014American Neurological Association

extensor hallucis longus,and extensor digitorum longus.1Clinical symptoms typically begin at age 35to 55years,and the disease is slowly progressive.1TMD was first described in Finnish patients;its prevalence in Finland is estimated to be >1=10,000,2which makes it the most common muscle disease in Finland.The Finnish founder mutation (FINmaj)is an 11bp insertion-deletion mutation exchanging 4amino acids in the 363rd and last exon of TTN (Mex6),which encodes the C-terminal immuno-globulin domain M10of M-line titin.3In addition to the FINmaj mutation,we have reported several other muta-tions causing TMD in different European populations.These are missense or truncating mutations located in the last 2TTN exons (Mex5and Mex63–6;Fig 1).

Homozygosity of the FINmaj mutation is rare but causes a completely different limb-girdle muscular dystro-phy phenotype,LGMD2J (OMIM #608807).7,8LGMD2J is a severe childhood onset disease causing proximal muscle weakness in the first or second decade and progresses over the next 20years to wheelchair con-finement.7,8One patient homozygous for the truncating French TMD mutation c.107890C >T (p.Q35964*;NM_001267550.1)has also been reported with a proxi-mal phenotype somewhat different from the homozygous Finnish LGMD2J patients.In the French patient,the disease onset was at 25years,and the first muscle weak-ness occurred in the proximal upper limbs.Weakness and wasting progressed to all 4limbs,and the patient lost ambulation at the age of 56years.9

TMD FINmaj mutations in the last M10domain of titin are predicted to cause cleavage of a larger part of the titin C-terminus,because immunofluorescence studies of homozygote LGMD2J muscles showed an absence of titin M8=M9domain epitopes.3Also,Western blot anal-ysis showed severe reduction of the C-terminal titin frag-ments in LGMD2J patient muscle.5Loss of protein interactions of C-terminal titin is thus a likely conse-quence of the TMD =LGMD2J mutations.Obscurin and obscurin-like 1are known to interact with the titin M10domain,and in LGMD2J muscles the FINmaj mutation leads to mislocalization of obscurin.10Next to the M10domain is the is7region,encoded by the second-last

exon 362(Mex5),which contains an M-line binding site for the muscle-specific protease calpain 3.11Calpain 3is severely reduced in LGMD2J,and it has been postulated that the LGMD2J phenotype is caused by a loss-of-function mechanism including secondary calpain 3abnormality,12,13in contrast to the dominant late onset effect of the FINmaj mutation in TMD heterozygous patients.

Several families and patients with previously reported TMD-causing TTN mutations have more com-plex,severe,or unusual phenotypes.5,8This study was initiated to clarify the molecular cause of the variant phe-notypes observed in 4Finnish and 3Southern European families.Five patients proved to be compound heterozy-gotes with novel titin frameshift mutations combined with the previously reported TMD mutations.One Por-tuguese patient was homozygous for the previously reported Iberian TMD mutation,and 1patient had the FINmaj mutation combined with a missense mutation in the A-band titin causing a new phenotype completely different from previously described titinopathies.

Subjects and Methods

Patients and Biopsies

DNA samples from 8patients of 7families were obtained from clinicians in different countries and screened for mutations in TTN .Four families were from Finland and 1each from France,Spain,and Portugal.Muscle biopsies were available for protein and mRNA studies in all probands.The study was approved by the local ethics committees,and samples were obtained accord-ing to the Helsinki declaration.

Production and Testing of Monoclonal Antibodies against the M10Domain

The M10domain of human titin was cloned into pGEX-6P-2(GE Healthcare,Little Chalfont,UK).The GST-M10was expressed in Escherichia coli BL21and affinity purified at Biomolecular Tools Finland Oy (Turku,Finland).The GST moiety was removed by cleavage with PreScission protease (GE Healthcare)and bound to Glutathione Sepharose resin.The M10domain was purified by gel filtration and concentrated with Amicon Ultra cartridge (Merck Millipore,Billerica,

MA).

FIGURE 1:Titin M-line.Tibial muscular dystrophy (TMD)mutations are located in Mex5and Mex6exons of TTN encoding the

is7region and the most C-terminal domain M10in the sarcomere M-line.Calpain 3has binding site in is7and obscurin,obscurin-like 1,and myospryn bind to M10.

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Monoclonal antibody development against the M10 domain conjugated to diphtheria toxin was carried out at Bio-Genes(Berlin,Germany).The hybridoma clones were screened by enzyme-linked immunosorbent assay against native and denatured immunogen and further tested by Western blotting of wild-type and mutant titin is6-M10constructs14expressed in COS-1cells,as well as muscle extracts.For the3clones selected for production,epitope mapping was performed on a SPOT array containing human and mouse M10sequences as 15-mer peptides with a tiling of3residues(Peptide Synthesis Unit,Haartman Institute,University of Helsinki).

C-Terminal M10Domain Monoclonal Titin Antibodies

Development of M10monoclonal antibodies resulted in3 clones(designated7-4-4,11-4-3,and14-2-7)that efficiently stained C-terminal titin constructs in Western blotting,and rec-ognized linear epitopes of human titin on peptide array.11-4-3 recognized the sequence QGRFHI in the M10domain of titin. The epitope is identical between human and mouse titin,and the antibody bound sequences from both species equally well on the peptide array.The specificity of the antibodies was fur-ther tested by Western blot using skeletal muscle lysates from healthy control,FINmaj heterozygous TMD,and FINmaj homozygous LGMD2J.All3antibodies detected several bands of various sizes,but only11-4-3showed specificity to titin C-terminus in skeletal muscle lysates,as demonstrated by loss of most bands in the LGMD2J sample.Only2bands(approxi-mate molecular weights530and100kDa)remained unchanged,suggesting that they represent unspecific cross reactions.

Western Blotting

Muscle biopsies were homogenized in reducing sample buffer containing4%sodium dodecyl sulfate(SDS)and10%b-mercaptoethanol,and heated for5minutes at100 C.SDS–polyacrylamide gel electrophoresis and Western blotting were performed according to standard methods.T wo different pri-mary antibodies raised against titin M10,rabbit polyclonal anti-body M10-15and mouse monoclonal antibody11-4-3,were used.In addition,mouse monoclonal calpain3antibody clones 2C4and12A2(Leica Biosystems,Newcastle Upon Tyne,UK) were used,followed by horseradish peroxidase–conjugated sec-ondary antibodies(Dako,Glostrup,Denmark)and enhanced chemiluminescence detection using the Pierce SuperSignal West Femto substrate(Thermo Fisher Scientific,Waltham,MA). RNA Extraction and Reverse Transcription Polymerase Chain Reaction

Total RNA was extracted with the RNeasy Fibrous Tissue Mini Kit(Qiagen,Hilden,Germany)according to the manufacturer’s instructions.cDNA was synthesized with the High Capacity cDNA Reverse T ranscription Kit(Applied Biosystems,Foster City,CA)using random hexanucleotides.Primers spanning exon–exon junctions were designed for cDNA with Primer3 software.Fragments were amplified with DreamTaq DNA Polymerase(Fermentas,Vilnius,Lithuania),separated in agarose gel,and isolated from gel for sequencing.

Sequencing

Sanger sequencing of genomic DNA and cDNA was performed with standard techniques.Primer sequences are listed in the Supplementary Table.Polymerase chain reaction(RT-PCR)was performed using DreamTaq DNA Polymerase(Fermentas) according to standard protocol.PCR products were sequenced on an ABI3730xl DNA Analyzer(Applied Biosystems),using the Big-Dye T erminator v3.1kit and analyzed with Sequencher 5.0software(Gene Codes Corporation,Ann Arbor,MI). Results

Clinical Characteristics of the Patients PATIENT1.A16-year-old Finnish female was known to be heterozygous for the FINmaj mutation,inherited from her mother.The mother was diagnosed at age50years with rather mild TMD;the father is healthy.The first symptoms occurred in early infancy,with slightly delayed walking and no stage of crawling.At the age of4years, she was referred because of a peculiar walking and short-ness of breath after walking100m.At the age of6years she was unwilling to walk and had difficulties in climbing stairs.She had large calves,external rotation of the lower limbs while walking,and increased lordosis.Muscle biopsy showed early dystrophic findings with rimmed vacuolar pathology,whereas the immunohistochemistry of sarco-lemmal membrane proteins was normal.Muscle magnetic resonance imaging(MRI)at age12years showed no defi-nite fatty replacement,but proximal muscle volume was decreased,and there was minor edema on short time inversion recovery sequences in the right rectus femoris (Fig2A).At age16years,she needs help in carrying her bag and getting up from the floor.She has mild contrac-tures in the ankles,knees,and elbows.

PATIENTS2AND 3.Patients2and3have been reported earlier.8Patient2was characterized as having infantile-onset generalized weakness and Patient3as hav-ing childhood onset limb-girdle muscular dystrophy. Muscle imaging of Patient3is shown in Figure2B. PATIENT4.A51-year-old Finnish male is heterozygous for the FINmaj mutation but with a phenotype differing completely from TMD,LGMD2J,or any other described titinopathy.The patient’s father is asymptom-atic and the mother has TMD with the FINmaj muta-tion.The first symptoms occurred at the age of30years with muscle atrophy in the right calf.Ten years later, muscle atrophy was observed also in the right thigh,pro-gressing later to the left thigh.The upper and left distal lower limbs remained normal.The patient started to use

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a cane after age 40years.Muscle biopsy from the right vastus lateralis showed end-stage level nonspecific dystro-phic changes,whereas a second biopsy from the left vastus lateralis showed pronounced rimmed-vacuolar pathology without myofibrillar myopathy pathology.MRI findings were unique,with fatty degenerative and

atrophic changes in soleus and quadriceps on both sides and total tissue replacement by fat and connective tissue in the right gastrocnemius medialis and lateralis muscles (see Fig 2C).In addition,similar less severe changes were observed in gluteus medius,adductor longus,and semite-ndinosus on the right

side.

FIGURE 2:Muscle imaging of the patients.(A)Patient 1at the age of 12years.The musculature is still relatively well pre-served.There is edema in the right rectus femoris muscle when compared to the left side (upper),but without dystrophic

degenerative change.The left medial head of gastrocnemius shows edema,and its volume is reduced compared to the right side.(B)Patient 3at the age of 30years.All muscles are severely affected.All hamstring and adductor muscles show variable degrees of severe fatty degeneration and atrophy.(C)Patient 4at the age of 51years.The quadriceps and soleus muscles show advanced dystrophic changes.At the right side also the semitendinosus,gastrocnemius medialis,and gastrocnemius lat-eralis show severe fatty degeneration and marked atrophy.Less severe changes are seen in the adductor longus muscle on the right.The tibialis anterior muscles are relatively spared.(D)Patient 5B at the age of 40years.Severe dystrophic fatty degener-ation is seen in the anterolateral compartments as well as soleus muscles of the lower legs.(E)Patient 6at the age of 26years.The posterior thigh muscles,particularly on the right side,show dystrophic changes.Most severely affected are the long head of biceps femoris,semitendinosus,and semimembranosus muscles.On the lower legs,the left tibialis anterior is already severely involved.The left gastrocnemius medialis is more severely affected than the right,and the soleus more on the right side.(F)Patient 7at the age of 25years.At the thigh level,biceps femoris (long head),semimembranosus,and semitendinosus show moderate fatty degeneration,and quadriceps shows early changes on the left side.Gracilis is hypertrophic,and biceps femoris (short head)is spared.The tibialis anterior,extensor hallucis longus,soleus,and peroneal muscles are severely dystro-phic,and the extensor digitorum longus is replaced more on the left side.The gastrocnemius medialis and lateralis muscles are hypertrophic.

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PATIENTS 5A AND 5B.Patients 5A and 5B (French Family C),have been reported earlier.5

Patient 5A is the mother of Patient 5B.Muscle imaging of Patient 5B is shown in Figure 2D.

PATIENT 6.The patient is a 36-year-old Spanish

female heterozygous for the previously reported Iberian TMD mutation c.107889delA (p.K35963Nfs*9).5She has an unusual early onset of the disease and a

relatively

FIGURE 3:Western blotting.(A)Western blotting of control skeletal muscle lysate with the titin C-terminal antibodies M10-1

and 11-4-3reveals a distinctive pattern of bands,representing C-terminal protein fragments of different lengths.The specific-ity of the bands is demonstrated in a FINmaj tibial muscular dystrophy (TMD)sample,which shows reduction of bands,and in FINmaj limb-girdle muscular dystrophy type 2J (LGMD2J),where the immunoreactivity is nearly completely lost.M10-1bands appear specific,whereas with 11-4-3,the bands of approximately 30and 90kDa probably represent unspecific cross-reactions with unknown proteins.Especially the samples from Patients 1,5A,6,and 7show a fragment pattern similar to LGMD2J,with dramatic loss of titin C-terminal epitope recognition.(B)Western blotting of calpain 3shows clear reduction of the full-size 90kDa calpain 3and its proteolytic fragments (30and 60kDa with 2C4and 12A2,respectively)in Patients 1,3,6,and 7.Cal-pain 3levels in Patient 5A appear normal,whereas Patient 4shows slight reduction possibly caused by degradation as seen by enhanced proteolytic fragment bands,and lower total protein loading.In addition,the lower sensitivity of 12A2antibody may account for the reduced 90kDa band.

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severe phenotype,especially compared to her heterozy-gous mother,who is still asymptomatic at the age of 58years.Motor milestones were slightly delayed with walk-ing after the age of 18months and general motor clum-siness during infancy.Walking difficulties and weakness slightly progressed,and the first neurological examination at the age of 25years showed distal weakness in ankle dorsiflexors and mild weakness of hip flexors and knee flexors in the lower limbs.Muscle MRI at the age of 26years displayed bilateral atrophy of soleus and tibialis anterior with more fatty replacement on the left side (see Fig 2E).The muscles in the posterior compartment of the thighs showed atrophy on both sides and fatty degen-eration more on the right side.

PATIENT 7.The patient,a 27-year-old Portuguese

female,has a more severe distal TMD phenotype,with early adult onset at 22years of age and slowly progressive difficulty in running and frequent tripping.Neurological examination showed bilateral atrophy of the anterolateral compartment of the legs and severe weakness of ankle dorsiflexion.Achilles tendon reflexes were abolished with mild Achilles contractures.Lower limb muscle MRI showed marked atrophy and fat infiltration of the antero-lateral compartment and soleus muscles of the lower legs (see Fig 2F).At the thigh level,hamstring muscles were severely replaced,as were gluteus medius and minimus of the pelvic region.The patient proved to be homozygous for the previously reported Iberian TMD mutation c.107889delA (p.K35963Nfs*9).5The parents,first-degree cousins,are heterozygous for the same mutation,without symptoms or clinical signs of muscle disease at the age of 55to 60years,although subclinical myopathy with smaller lesions of fatty degeneration in gastrocne-mius and soleus muscles were present on MRI of the father.

Patients’clinical characteristics are summarized in Table 1.

Molecular Biological Findings

Titin and calpain 3proteins were studied from muscle samples of patients by Western blotting.Reduction of C-terminal titin protein fragments of <200kDa was observed in all analyzed patients,and a complete loss of C-terminal antigen recognition was seen in Patients 1,2,5A,5B,6,and 7,corresponding to the pattern seen in FINmaj homozygous LGMD2J (Fig 3A).Calpain 3showed clear reduction in Patients 1,3,6,and 7,but was within normal range in Patients 4and 5A (see Fig 3B).Normal CAPN3transcript in Patients 1and 3,ana-lyzed by cDNA sequencing,indicated that calpain 3pro-tein deficiency was secondary,as reported previously in the case of FINmaj homozygous LGMD2J.5

Expression levels of the TTN alleles were studied by RT-PCR from muscle biopsies of Patients 1,3,5A,and 5B following RNA extraction and cDNA synthesis.A region spanning exons Mex4–Mex6or Mex5–Mex6was amplified from the cDNA and sequenced.Sequenc-ing chromatograms indicated unequal expression of the 2TTN alleles,with lower signals from the other alleles compared to the alleles containing the previously known TMD mutation (Fig 4).These Western blotting and RT-PCR results indicated that the other TTN allele might contain a frameshift mutation affecting its expression.

In search of additional mutations,genomic DNA of the patients was sequenced starting from the last exon of TTN and continuing upstream.A single-nucleotide deletion c.101113delT was identified in exon 358

of

FIGURE 4:cDNA sequencing chromatograms of the TTN Mex6FINmaj tibial muscular dystrophy (TMD)region of a wild-type (WT)subject,a FINmaj TMD patient,and Patient 1.The chromatograms indicate unequal expression of the 2TTN alleles in Patient 1,with lower expression of the other allele compared to the FINmaj TMD allele.

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Patient1,a single-nucleotide duplication c.39492dupT in exon207of Patient2,a single-nucleotide deletion c.98105delC in exon352of Patient5A,a4-nucleotide duplication c.100558-100561dupACTG in exon357of Patient5B,and a single-nucleotide deletion c.67089delT in exon318of Patient6,all5causing frameshifts and premature stop codons.Patient4had,in addition to the FINmaj mutation,a missense mutation c.92167C>T (p.P30723S)in TTN exon339.All these mutations were novel.Patient7showed the previously reported Iberian TMD mutation c.107889delA(p.K35963Nfs*9)homo-zygously.All found mutations and their consequences on protein level are summarized in Table2.All363TTN exons from Patient3were also sequenced,but no addi-tional mutation was identified with the primers used. Sequencing the DNA from both parents of Patients1,2, 4,and6,and from the mother of Patient5B confirmed that the new mutations were located on different alleles than the previously found and reported TMD mutations.

Subsequently,RT-PCR and sequencing of cDNA from muscle biopsy were performed with primers designed for the TTN regions containing the new muta-tions found in Patients1,2,5A,5B,and 6.The sequencing chromatograms indicated a lower signal from the frameshift mutated alleles compared to the TMD alleles,thus confirming earlier results of RT-PCR of exons Mex4–Mex6.

Discussion

Five of the studied patients with complex TMD pheno-types turned out to be compound heterozygotes,each with1previously known TMD mutation and5different novel frameshift mutations.Based on sequencing chro-matograms of RT-PCR products from Patients1,2,5A, 5B,and6,there was in each case an indication of a much lower expression of the TTN allele with the frame-shift mutation compared to the other allele with previ-ously known TMD mutation.All the novel frameshift mutations cause a premature stop codon in the sequence more than50to55nucleotides upstream of the last exon–exon junction,which is likely to trigger nonsense-mediated decay(NMD).15When the frameshift mutated mRNAs are degraded,the other TTN alleles with the known TMD mutations will be predominantly expressed. This may lead to compound heterozygous patients hav-ing a more severe phenotype or phenotypes similar to those homozygous for the corresponding TMD muta-tion.Because NMD is usually not complete,small amounts of truncated protein may still be expressed.As expected,the frameshift mutated alleles are recessive and alone do not cause disease;the fathers of Patients1and 6and the mother of Patient2,carriers of the frameshift alleles,were healthy.

A second mutation has not been found from Patient3,although all TTN exons have been sequenced. It is,however,likely that also this patient has a frameshift mutation in the other TTN allele,as there is loss of C-terminal epitopes on Western blotting and the RT-PCR results resemble those of Patients1,2,5A,5B,and6.A possible explanation for this putative second mutation escaping detection could be that the second mutation is a bigger frameshift-causing deletion or an inversion not detectable with Sanger sequencing,or that the primers used just do not identify the mutation despite showing good normal sequence.Other possibilities would include a deeper intronic mutation causing aberrant splicing,a mutation in the promoter or enhancer region of TTN, or an epigenetic change decreasing expression of the allele.

Loss of titin C-terminus in Western blotting in samples of Patients1,2,5A,5B,6,and7was indistin-guishable from that seen in LGMD2J.This is in line with the predominant expression of the TMD alleles due to NMD of the other allele,leading to loss of functional C-terminal M10domain at the protein level.Patient3 showed residual staining of a high molecular weight titin band with the11-4-3monoclonal antibody,although the smaller C-terminal titin fragments were missing.The case of Patient4confirms the conclusion above,as the combination of the FINmaj and the novel A-band titin missense mutation shows less severe loss of titin C-terminus as compared to the FINmaj combined with a frameshift mutation(see Fig3).

Surprisingly,calpain3protein appeared normal in Patient5A,with the Mex5TMD mutation,in contrast to Patients1,3,6,and7,who showed variable but clear calpain3reduction.Patient4showed a slightly reduced 94kDa calpain3band,but proteolytic fragments were more pronounced,suggesting mild sample degradation that together with slightly lower total protein loading indicates that calpain3is within normal range.Previ-ously all FINmaj LGMD2J patients have shown severe reduction of calpain3,whereas in heterozygous FINmaj TMD patients the amount of calpain3has been variably normal.

The Iberian TMD mutation c.107889delA (p.K35963Nfs*9)appears to have a reduced penetrance or recessive properties,because the parents of Patient7 and the mother of Patient6are heterozygous for the mutation but seem to be subjectively healthy at age55 to60years.It is also possible that they have a very late disease onset,as indicated by minor changes on muscle MRI in the father of Patient7.

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Patient7,homozygous for the Iberian TMD muta-tion,and the compound heterozygous Patients5A,5B and 6,have a more severe TMD phenotype rather than LGMD2J phenotype seen in FINmaj homozygous patients and in Patients1and2with FINmaj combined with a fra-meshift mutation.The reason why some homozygous TMD mutations or compound heterozygous TMD with frameshift mutation cause a more severe TMD,whereas others cause LGMD2J,is currently not known.The Ibe-rian TMD mutation is located C-terminally from the FIN-maj mutation,very close to titin C-terminus,and could thus have a milder effect on the domain structure.How-ever,1patient was recently reported to have the Iberian TMD mutation combined with a nonsense mutation and causing an LGMD2J phenotype.16Also a patient homozy-gous for the French Mex6TMD mutation c.107890C>T (p.Q35964*),located next to the Iberian mutation and thus expected to have a comparable effect,showed a pheno-type that resembled LGMD2J but also included involve-ment of distal muscles.9

The TMD phenotypes in Patients5A and5B are interesting.The patients carry,in addition to the French Mex5mutation,2different second mutations severely decreasing the expression level of the other allele on the mRNA level.The clear reduction of C-terminal titin,con-firmed by Western blotting,is consistent with functional homozygosity for the Mex5mutation;yet the patients show a severe TMD phenotype rather than LGMD2J. One factor contributing to the phenotype in both patients of this family could be the alternative splicing of Mex5;as the Mex52(is72)isoforms skip the mutation,C-terminal titin could actually retain some of its function,explaining why the patients do not develop the LGMD2J phenotype. However,the titin M10immunoreactivity in the Western blot was very weak,and did not reveal significant amounts of normal Mex52titin in the analyzed biopsies.

Mex5encodes the is7region containing a binding site for calpain3.Surprisingly,Western blotting did not indicate a reduction of calpain3in Patient5A,with the Mex5TMD mutation,in contrast to patients with Mex6 TMD mutations.We have previously postulated that in the case of FINmaj mutation,the loss of calpain3bind-ing could be due to proteolytic cleavage of titin upstream of the domains M8=M9,because LGMD2J(FINmaj homozygote)muscles are missing immunoreactivity to the M8=M9domain antibody3and they also have severely reduced calpain3.12As the French Mex5TMD mutation truncates the is7region C-terminally,close to the is7=M10boundary,it is possible that the mutation does not directly interfere with the binding site of cal-pain 3.Moreover,because the mutation removes the entire M10without causing domain unfolding,the mutant protein might be less prone to proteolytic cleav-age,thus providing adequate sites for calpain3binding.

Patient4had the FINmaj mutation combined with a missense mutation c.92167C>T(p.P30723S)that changes a proline to a serine in the A140domain of A-band titin. The patient has inherited the A-band mutation from his healthy father,proving its recessive properties.Thus far, reported pathogenic mutations in the A-band titin are mis-sense mutations in the A150domain causing hereditary myopathy with early respiratory failure,17–21missense mutations in different A-band regions causing arrhythmo-genic right ventricular cardiomyopathy,22and truncating mutations in different A-band regions causing dilated car-diomyopathy.23,24The phenotype of the patient,however, differs completely from previously described titinopathies. Although the patient has the FINmaj mutation,the symp-toms are more severe and not similar to either TMD or LGMD2J.He has marked dystrophic changes in quadri-ceps femoris and soleus,but tibialis anterior has a normal appearance on MRI.So far,we do not have a plausible explanation for how this compound heterozygosity gives rise to the observed new titinopathy phenotype.

Frameshift and nonsense mutations in different A-band regions of titin have previously been reported to cause dilated cardiomyopathy.23,24The mutations may cause expression of truncated titin proteins,which are integrated into the sarcomere and cause disease by means of a domi-nant negative mechanism.23T runcated titin proteins are also expressed in patients with early onset myopathy with fatal cardiomyopathy25and in patients with core myopa-thy26who have homozygous frameshift mutations in the M-line titin.None of the patients in our study showed car-diomyopathy.Instead,the identified frameshift mutations in the compound heterozygous patients of our study very likely cause NMD with no or only small amounts of pro-tein product expressed from the frameshift mutated allele.

Our results considerably expand the range of mus-cle disorders caused by TTN mutations and suggest that the coexistence of second mutations may be a more com-mon general mechanism causing phenotypic variability. Many muscular dystrophy phenotype variations are known to exist in patients with identical causative muta-tions,and our study may provide new insight into an underlying mechanism of those variations. Acknowledgment

This study was supported by the Folkh€a lsan Research Foundation,the Association Franc?aise contreles Myopa-thies(SB/CP2013-0106,B.U.),the Academy of Finland (no.138491, B.U.),the Sigrid Jus e lius Foundation (B.U.),the Liv och H€a lsa Foundation(P.H., B.U.),

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Tampere University Hospital Research Funds(B.U.),the Alfred Kordelin Foundation(J.S.),and the Orion-Farmos Research Foundation(J.S.).

We thank M.Soininen,H.Luque,J.Leppikangas, and S.Luhtasela for technical assistance.

Authorship

A.E.,A.V.:acquisition of data,analysis and interpretation of data,drafting=revising manuscript.J.S.:analysis and interpretation of data,drafting=revising manuscript. O.R.:acquisition of data,analysis and interpretation of data,revising manuscript.J.P.:analysis and interpretation of data,drafting=revising manuscript.S.S.,

B.E.,I.I., R.R.-G.,K.H.,L.N.,T.L.,P.N.,M.K.,S.P.,M.S.,T.S., I.R.:acquisition of data,analysis and interpretation of data,revising manuscript.P.H.:acquisition of data,anal-ysis and interpretation of data,drafting=revising manu-script,study supervision. B.U.:study conception and design,acquisition of data,analysis and interpretation of data,drafting=revising manuscript,study supervision. Potential Conflicts of Interest

R.R.-G.:grants=grants pending,FIS;paid advisory board,Pfizer.P.N.:speaking fees,Glaxo-Smith-Kline Fin-land;travel expenses,Novartis.

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ANNALS of Neurology

240Volume75,No.2

常用二极管参数

常用整流二极管 型号VRM/Io IFSM/ VF /Ir 封装用途说明1A5 600V/1.0A 25A/1.1V/5uA[T25] D2.6X3.2d0.65 1A6 800V/1.0A 25A/1.1V/5uA[T25] D2.6X3.2d0.65 6A8 800V/6.0A 400A/1.1V/10uA[T60] D9.1X9.1d1.3 1N4002 100V/1.0A 30A/1.1V/5uA[T75] D2.7X5.2d0.9 1N4004 400V/1.0A 30A/1.1V/5uA[T75] D2.7X5.2d0.9 1N4006 800V/1.0A 30A/1.1V/5uA[T75] D2.7X5.2d0.9 1N4007 1000V/1.0A 30A/1.1V/5uA[T75] D2.7X5.2d0.9 1N5398 800V/1.5A 50A/1.4V/5uA[T70] D3.6X7.6d0.9 1N5399 1000V/1.5A 50A/1.4V/5uA[T70] D3.6X7.6d0.9 1N5402 200V/3.0A 200A/1.1V/5uA[T105] D5.6X9.5d1.3 1N5406 600V/3.0A 200A/1.1V/5uA[T105] D5.6X9.5d1.3 1N5407 800V/3.0A 200A/1.1V/5uA[T105] D5.6X9.5d1.3 1N5408 1000V/3.0A 200A/1.1V/5uA[T105] D5.6X9.5d1.3 RL153 200V/1.5A 60A/1.1V/5uA[T75] D3.6X7.6d0.9 RL155 600V/1.5A 60A/1.1V/5uA[T75] D3.6X7.6d0.9 RL156 800V/1.5A 60A/1.1V/5uA[T75] D3.6X7.6d0.9 RL203 200V/2.0A 70A/1.1V/5uA[T75] D3.6X7.6d0.9 RL205 600V/2.0A 70A/1.1V/5uA[T75] D3.6X7.6d0.9 RL206 800V/2.0A 70A/1.1V/5uA[T75] D3.6X7.6d0.9 RL207 1000V/2.0A 70A/1.1V/5uA[T75] D3.6X7.6d0.9 RM11C 1000V/1.2A 100A/0.92V/10uA D4.0X7.2d0.78 MR750 50V/6.0A 400A/1.25V/25uA D8.7x6.3d1.35 MR751 100V/6.0A 400A/1.25V/25uA D8.7x6.3d1.35 MR752 200V/6.0A 400A/1.25V/25uA D8.7x6.3d1.35 MR754 400V/6.0A 400A/1.25V/25uA D8.7x6.3d1.35 MR756 600V/6.0A 400A/1.25V/25uA D8.7x6.3d1.35 MR760 1000V/6.0A 400A/1.25V/25uA D8.7x6.3d1.35 常用整流二极管(全桥） 型号VRM/Io IFSM/ VF /Ir 封装用途说明RBV-406 600V/*4A 80A/1.10V/10uA 25X15X3.6 RBV-606 600V/*6A 150A/1.05V/10uA 30X20X3.6 RBV-1306 600V/*13A 80A/1.20V/10uA 30X20X3.6 RBV-1506 600V/*15A 200A/1.05V/50uA 30X20X3.6 RBV-2506 600V/*25A 350A/1.05V/50uA 30X20X3.6 常用肖特基整流二极管SBD 型号VRM/Io IFSM/ VF Trr1/Trr2 封装用途说明EK06 60V/0.7A 10A/0.62V 100nS D2.7X5.0d0.6 SK/高速 EK14 40V/1.5A 40A/0.55V 200nS D4.0X7.2d0.78 SK/低速 D3S6M 60V/3.0A 80A/0.58V 130p SB340 40V/3.0A 80A/0.74V 180p SB360 60V/3.0A 80A/0.74V 180p SR260 60V/2.0A 50A/0.70V 170p MBR1645 45V/16A 150A/0.65V <10nS TO220 超高速

常用二极管参数

常用二极管参数 2008-10-22 11:48 05Z6.2Y 硅稳压二极管 Vz=6~6.35V, Pzm=500mW, 05Z7.5Y 硅稳压二极管 Vz=7.34~7.70V, Pzm=500mW, 05Z13X 硅稳压二极管 Vz=12.4~13.1V, Pzm=500mW, 05Z15Y 硅稳压二极管 Vz=14.4~15.15V, Pzm=500mW, 05Z18Y 硅稳压二极管 Vz=17.55~18.45V, Pzm=500mW, 1N4001 硅整流二极管 50V, 1A,(Ir=5uA, Vf=1V, Ifs=50A) 1N4002 硅整流二极管 100V, 1A, 1N4003 硅整流二极管 200V, 1A, 1N4004 硅整流二极管 400V, 1A, 1N4005 硅整流二极管 600V, 1A, 1N4006 硅整流二极管 800V, 1A, 1N4007 硅整流二极管 1000V, 1A, 1N4148 二极管 75V, 4PF, Ir=25nA, Vf=1V, 1N5391 硅整流二极管 50V, 1.5A,(Ir=10uA, Vf=1.4V, Ifs=50A) 1N5392 硅整流二极管 100V, 1.5A, 1N5393 硅整流二极管 200V, 1.5A, 1N5394 硅整流二极管 300V, 1.5A, 1N5395 硅整流二极管 400V, 1.5A, 1N5396 硅整流二极管 500V, 1.5A, 1N5397 硅整流二极管 600V, 1.5A, 1N5398 硅整流二极管 800V, 1.5A, 1N5399 硅整流二极管 1000V, 1.5A, 1N5400 硅整流二极管 50V, 3A,(Ir=5uA, Vf=1V, Ifs=150A) 1N5401 硅整流二极管 100V, 3A, 1N5402 硅整流二极管 200V, 3A, 1N5403 硅整流二极管 300V, 3A, 1N5404 硅整流二极管 400V, 3A, 1N5405 硅整流二极管 500V, 3A, 1N5406 硅整流二极管 600V, 3A, 1N5407 硅整流二极管 800V, 3A, 1N5408 硅整流二极管 1000V, 3A, 1S1553 硅开关二极管 70V, 100mA, 300mW, 3.5PF, 300ma, 1S1554 硅开关二极管 55V, 100mA, 300mW, 3.5PF, 300ma, 1S1555 硅开关二极管 35V, 100mA, 300mW, 3.5PF, 300ma, 1S2076 硅开关二极管 35V, 150mA, 250mW, 8nS, 3PF, 450ma, Ir≤1uA, Vf≤0.8V,≤1.8PF, 1S2076A 硅开关二极管 70V, 150mA, 250mW, 8nS, 3PF, 450ma, 60V, Ir≤1uA, Vf≤0.8V,≤1.8PF, 1S2471 硅开关二极管80V, Ir≤0.5uA, Vf≤1.2V,≤2PF, 1S2471B 硅开关二极管 90V, 150mA, 250mW, 3nS, 3PF, 450ma, 1S2471V 硅开关二极管 90V, 130mA, 300mW, 4nS, 2PF, 400ma, 1S2472 硅开关二极管50V, Ir≤0.5uA, Vf≤1.2V,≤2PF, 1S2473 硅开关二极管35V, Ir≤0.5uA, Vf≤1.2V,≤3PF,

joinin剑桥小学英语

Join In剑桥小学英语(改编版)入门阶段Unit 1Hello,hello第1单元嗨,嗨 and mime. 1 听,模仿 Stand up Say 'hello' Slap hands Sit down 站起来说"嗨" 拍手坐下来 Good. Let's do up Say 'hello' Slap hands Sit down 好. 我们来再做一遍.站起来说"嗨"拍手坐下来 the pictures. 2 再听一遍给图画编号. up "hello" hands down 1 站起来 2 说"嗨" 3 拍手 4 坐下来说 3. A ,what's yourname 3 一首歌嗨,你叫什么名字 Hello. , what's yourname Hello. Hello. 嗨. 嗨. 嗨, 你叫什么名字嗨,嗨. Hello, what's yourname I'm Toby. I'm Toby. Hello,hello,hello.嗨, 你叫什么名字我叫托比. 我叫托比 . 嗨,嗨,嗨. I'm Toby. I'm Toby. Hello,hello, let's go! 我是托比. 我是托比. 嗨,嗨, 我们一起! Hello. , what's yourname I'm 'm Toby. 嗨.嗨.嗨, 你叫什么名字我叫托比.我叫托比. Hello,hello,hello. I'm 'm Toby. Hello,hello,let's go! 嗨,嗨,嗨.我是托比. 我是托比. 嗨,嗨,我们一起! 4 Listen and stick 4 听和指 What's your name I'm Bob. 你叫什么名字我叫鲍勃. What's your name I'm Rita. What's your name I'm Nick. 你叫什么名字我叫丽塔. 你叫什么名字我叫尼克. What's your name I'm Lisa. 你叫什么名字我叫利萨. 5. A story-Pit'sskateboard. 5 一个故事-彼德的滑板. Pa:Hello,Pit. Pa:好,彼德. Pi:Hello,:What's this Pi:嗨,帕特.Pa:这是什么 Pi:My new :Look!Pi:Goodbye,Pat! Pi:这是我的新滑板.Pi:看!Pi:再见,帕特! Pa:Bye-bye,Pit!Pi:Help!Help!pi:Bye-bye,skateboard! Pa:再见,彼德!Pi:救命!救命!Pi:再见,滑板! Unit 16. Let's learnand act 第1单元6 我们来边学边表演.

常用二极管型号及参数大全精编版

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常用稳压二极管大全,

常用稳压管型号对照——（朋友发的） 美标稳压二极管型号 1N4727 3V0 1N4728 3V3 1N4729 3V6 1N4730 3V9 1N4731 4V3 1N4732 4V7 1N4733 5V1 1N4734 5V6 1N4735 6V2 1N4736 6V8 1N4737 7V5 1N4738 8V2 1N4739 9V1 1N4740 10V 1N4741 11V 1N4742 12V 1N4743 13V 1N4744 15V 1N4745 16V 1N4746 18V 1N4747 20V 1N4748 22V 1N4749 24V 1N4750 27V 1N4751 30V 1N4752 33V 1N4753 36V 1N4754 39V 1N4755 43V 1N4756 47V 1N4757 51V 需要规格书请到以下地址下载， 经常看到很多板子上有M记的铁壳封装的稳压管，都是以美标的1N系列型号标识的，没有具体的电压值，刚才翻手册查了以下3V至51V的型号与电压的对 照值，希望对大家有用 1N4727 3V0 1N4728 3V3 1N4729 3V6 1N4730 3V9

1N4733 5V1 1N4734 5V6 1N4735 6V2 1N4736 6V8 1N4737 7V5 1N4738 8V2 1N4739 9V1 1N4740 10V 1N4741 11V 1N4742 12V 1N4743 13V 1N4744 15V 1N4745 16V 1N4746 18V 1N4747 20V 1N4748 22V 1N4749 24V 1N4750 27V 1N4751 30V 1N4752 33V 1N4753 36V 1N4754 39V 1N4755 43V 1N4756 47V 1N4757 51V DZ是稳压管的电器编号，是和1N4148和相近的，其实1N4148就是一个0.6V的稳压管，下面是稳压管上的编号对应的稳压值，有些小的稳压管也会在管体 上直接标稳压电压，如5V6就是5.6V的稳压管。 1N4728A 3.3 1N4729A 3.6 1N4730A 3.9 1N4731A 4.3 1N4732A 4.7 1N4733A 5.1 1N4734A 5.6 1N4735A 6.2 1N4736A 6.8 1N4737A 7.5 1N4738A 8.2 1N4739A 9.1 1N4740A 10 1N4741A 11 1N4742A 12 1N4743A 13

Join In剑桥小学英语.doc

Join In剑桥小学英语(改编版)入门阶段 Unit 1Hello,hello第1单元嗨,嗨 1.Listen and mime. 1 听,模仿 Stand up Say 'hello' Slap hands Sit down 站起来说"嗨" 拍手坐下来 Good. Let's do itagain.Stand up Say 'hello' Slap hands Sit down 好. 我们来再做一遍.站起来说"嗨"拍手坐下来 2.listen again.Number the pictures. 2 再听一遍给图画编号. 1.Stand up 2.Say "hello" 3.Slap hands 4.Sit down 1 站起来 2 说"嗨" 3 拍手 4 坐下来说 3. A song.Hello,what's yourname? 3 一首歌嗨,你叫什么名字? Hello. Hello.Hello, what's yourname? Hello. Hello. 嗨. 嗨. 嗨, 你叫什么名字? 嗨,嗨. Hello, what's yourname? I'm Toby. I'm Toby. Hello,hello,hello. 嗨, 你叫什么名字? 我叫托比. 我叫托比 . 嗨,嗨,嗨. I'm Toby. I'm Toby. Hello,hello, let's go! 我是托比. 我是托比. 嗨,嗨, 我们一起! Hello. Hello.Hello, what's yourname? I'm Toby.I'm Toby. 嗨.嗨.嗨, 你叫什么名字? 我叫托比.我叫托比. Hello,hello,hello. I'm Toby.I'm Toby. Hello,hello,let's go! 嗨,嗨,嗨.我是托比. 我是托比. 嗨,嗨,我们一起! 4 Listen and stick 4 听和指 What's your name? I'm Bob. 你叫什么名字? 我叫鲍勃. What's your name ? I'm Rita. What's your name ? I'm Nick. 你叫什么名字? 我叫丽塔. 你叫什么名字? 我叫尼克. What's your name ? I'm Lisa. 你叫什么名字? 我叫利萨. 5. A story-Pit'sskateboard. 5 一个故事-彼德的滑板. Pa:Hello,Pit. Pa:好,彼德. Pi:Hello,Pat.Pa:What's this? Pi:嗨,帕特.Pa:这是什么? Pi:My new skateboard.Pi:Look!Pi:Goodbye,Pat! Pi:这是我的新滑板.Pi:看!Pi:再见,帕特! Pa:Bye-bye,Pit!Pi:Help!Help!pi:Bye-bye,skateboard! Pa:再见,彼德!Pi:救命!救命!Pi:再见,滑板! Unit 16. Let's learnand act 第1单元6 我们来边学边表演.

二极管封装大全

二极管封装大全 篇一：贴片二极管型号、参数 贴片二极管型号.参数查询 1、肖特基二极管SMA（DO214AC） 2010-2-2 16:39:35 标准封装： SMA 2010 SMB 2114 SMC 3220 SOD123 1206 SOD323 0805 SOD523 0603 IN4001的封装是1812 IN4148的封装是1206 篇二：常见贴片二极管三极管的封装 常见贴片二极管/三极管的封装 常见贴片二极管/三极管的封装 二极管： 名称尺寸及焊盘间距其他尺寸相近的封装名称 SMC SMB SMA SOD-106 SC-77A SC-76/SC-90A SC-79 三极管： LDPAK

DPAK SC-63 SOT-223 SC-73 TO-243/SC-62/UPAK/MPT3 SC-59A/SOT-346/MPAK/SMT3 SOT-323 SC-70/CMPAK/UMT3 SOT-523 SC-75A/EMT3 SOT-623 SC-89/MFPAK SOT-723 SOT-923 VMT3 篇三：常用二极管的识别及ic封装技术 常用晶体二极管的识别 晶体二极管在电路中常用“D”加数字表示，如： D5表示编号为5的二极管。 1、作用：二极管的主要特性是单向导电性，也就是在正向电压的作用下，导通电阻很小;而在反向电压作用下导通电阻极大或无穷大。正因为二极管具有上述特性，无绳电话机中常把它用在整流、隔离、稳压、极性保护、编码控制、调频调制和静噪等电路中。 电话机里使用的晶体二极管按作用可分为：整流二极管(如1N4004)、隔离二极管(如1N4148)、肖特基二极管(如BAT85)、发光二极管、稳压二极管等。 2、识别方法：二极管的识别很简单，小功率二极管的N极(负极)，在二极管外表大多采用一种色圈标出来，有些二极管也用二极管专用符号来表示P极(正极)或N极(负极)，也有采用符号标志为“P”、“N”来确定二极管极性的。发光二极管的正负极可从引脚长短来识别，长

1N系列常用整流二极管的主要参数

1N 系列常用整流二极管的主要参数
反向工作 峰值电压 URM/V 额定正向 整流电流 整流电流 IF/A 正向不重 复浪涌峰 值电流 IFSM/A 正向 压降 UF/V 反向 电流 IR/uA 工作 频率 f/KHZ 外形 封装
型 号
1N4000 1N4001 1N4002 1N4003 1N4004 1N4005 1N4006 1N4007 1N5100 1N5101 1N5102 1N5103 1N5104 1N5105 1N5106 1N5107 1N5108 1N5200 1N5201 1N5202 1N5203 1N5204 1N5205 1N5206 1N5207 1N5208 1N5400 1N5401 1N5402 1N5403 1N5404 1N5405 1N5406 1N5407 1N5408
25 50 100 200 400 600 800 1000 50 100 200 300 400 500 600 800 1000 50 100 200 300 400 500 600 800 1000 50 100 200 300 400 500 600 800 1000
1
30
≤1
＜5
3
DO-41
1．5
75
≤1
＜5
3
DO-15
2
100
≤1
＜10
3
3
150
≤0.8
＜10
3
DO-27
常用二极管参数： 05Z6.2Y 硅稳压二极管 Vz=6~6.35V,Pzm=500mW,

剑桥小学英语Join_In

《剑桥小学英语Join In ——Book 3 下学期》教材教法分析2012-03-12 18:50:43| 分类：JOIN IN 教案| 标签：|字号大中小订阅. 一、学情分析： 作为毕业班的学生，六年级的孩子在英语学习上具有非常显著的特点： 1、因为教材的编排体系和课时不足，某些知识学生已遗忘，知识回生的现象很突出。 2、有的学生因受学习习惯及学习能力的制约，有些知识掌握较差，学生学习个体的差异性，学习情况参差不齐、两级分化的情况明显，对英语感兴趣的孩子很喜欢英语，不喜欢英语的孩子很难学进去了。 3、六年级的孩子已经进入青春前期，他们跟三、四、五年级的孩子相比已经有了很大的不同。他们自尊心强，好胜心强，集体荣誉感也强，有自己的评判标准和思想，对知识的学习趋于理性化，更有自主学习的欲望和探索的要求。 六年级学生在英语学习上的两极分化已经给教师的教学带来很大的挑战，在教学中教师要注意引导学生调整学习方式： 1、注重培养学生自主学习的态度 如何抓住学习课堂上的学习注意力，吸引他们的视线，保持他们高涨的学习激情，注重过程的趣味化和学习内容的简易化。 2、给予学生独立思考的空间。 3、鼓励学生坚持课前预习、课后复习的好习惯。 六年级教材中的单词、句子量比较多，难点也比较多，学生课前回家预习，不懂的地方查英汉词典或者其它资料，上课可以达到事半功倍的效果，课后复习也可以很好的消化课堂上的内容。 4、注意培养学生合作学习的习惯。 5、重在培养学生探究的能力：学习内容以问题的方式呈现、留给学生更多的发展空间。 二、教材分析： （一）.教材特点： 1．以学生为主体，全面提高学生的素质。

常见二极管参数大全

1N系列稳压管

快恢复整流二极管

常用整流二极管型号和参数 05Z6.2Y 硅稳压二极管 Vz=6~6.35V,Pzm=500mW, 05Z7.5Y 硅稳压二极管 Vz=7.34~7.70V,Pzm=500mW, 05Z13X硅稳压二极管 Vz=12.4~13.1V,Pzm=500mW, 05Z15Y硅稳压二极管 Vz=14.4~15.15V,Pzm=500mW, 05Z18Y硅稳压二极管 Vz=17.55~18.45V,Pzm=500mW, 1N4001硅整流二极管 50V, 1A,(Ir=5uA,Vf=1V,Ifs=50A) 1N4002硅整流二极管 100V, 1A, 1N4003硅整流二极管 200V, 1A, 1N4004硅整流二极管 400V, 1A, 1N4005硅整流二极管 600V, 1A, 1N4006硅整流二极管 800V, 1A, 1N4007硅整流二极管 1000V, 1A, 1N4148二极管 75V, 4PF,Ir=25nA,Vf=1V, 1N5391硅整流二极管 50V, 1.5A,(Ir=10uA,Vf=1.4V,Ifs=50A) 1N5392硅整流二极管 100V,1.5A, 1N5393硅整流二极管 200V,1.5A, 1N5394硅整流二极管 300V,1.5A, 1N5395硅整流二极管 400V,1.5A, 1N5396硅整流二极管 500V,1.5A, 1N5397硅整流二极管 600V,1.5A, 1N5398硅整流二极管 800V,1.5A, 1N5399硅整流二极管 1000V,1.5A, 1N5400硅整流二极管 50V, 3A,(Ir=5uA,Vf=1V,Ifs=150A) 1N5401硅整流二极管 100V,3A, 1N5402硅整流二极管 200V,3A, 1N5403硅整流二极管 300V,3A, 1N5404硅整流二极管 400V,3A,

常用稳压二极管技术参数及老型号代换.

常用稳压二极管技术参数及老型号代换 型号最大功耗 (mW) 稳定电压(V) 电流(mA) 代换型号国产稳压管日立稳压管 HZ4B2 500 3.8 4.0 5 2CW102 2CW21 4B2 HZ4C1 500 4.0 4.2 5 2CW102 2CW21 4C1 HZ6 500 5.5 5.8 5 2CW103 2CW21A 6B1 HZ6A 500 5.2 5.7 5 2CW103 2CW21A HZ6C3 500 6 6.4 5 2CW104 2CW21B 6C3 HZ7 500 6.9 7.2 5 2CW105 2CW21C HZ7A 500 6.3 6.9 5 2CW105 2CW21C HZ7B 500 6.7 7.3 5 2CW105 2CW21C HZ9A 500 7.7 8.5 5 2CW106 2CW21D HZ9CTA 500 8.9 9.7 5 2CW107 2CW21E HZ11 500 9.5 11.9 5 2CW109 2CW21G HZ12 500 11.6 14.3 5 2CW111 2CW21H HZ12B 500 12.4 13.4 5 2CW111 2CW21H HZ12B2 500 12.6 13.1 5 2CW111 2CW21H 12B2 HZ18Y 500 16.5 18.5 5 2CW113 2CW21J HZ20-1 500 18.86 19.44 2 2CW114 2CW21K HZ27 500 27.2 28.6 2 2CW117 2CW21L 27-3 HZT33-02 400 31 33.5 5 2CW119 2CW21M RD2.0E(B) 500 1.88 2.12 20 2CW100 2CW21P 2B1 RD2.7E 400 2.5 2.93 20 2CW101 2CW21S RD3.9EL1 500 3.7 4 20 2CW102 2CW21 4B2 RD5.6EN1 500 5.2 5.5 20 2CW103 2CW21A 6A1 RD5.6EN3 500 5.6 5.9 20 2CW104 2CW21B 6B2 RD5.6EL2 500 5.5 5.7 20 2CW103 2CW21A 6B1 RD6.2E(B) 500 5.88 6.6 20 2CW104 2CW21B RD7.5E(B) 500 7.0 7.9 20 2CW105 2CW21C RD10EN3 500 9.7 10.0 20 2CW108 2CW21F 11A2 RD11E(B) 500 10.1 11.8 15 2CW109 2CW21G RD12E 500 11.74 12.35 10 2CW110 2CW21H 12A1 RD12F 1000 11.19 11.77 20 2CW109 2CW21G RD13EN1 500 12 12.7 10 2CW110 2CW21H 12A3 RD15EL2 500 13.8 14.6 15 2CW112 2CW21J 12C3 RD24E 400 22 25 10 2CW116 2CW21H 24-1

剑桥小学英语join in五年级测试卷

五 年 级 英 语 测 试 卷 学校 班级 姓名 听力部分（共20分） 一、Listen and colour . 听数字，涂颜色。（5分） 二、 Listen and tick . 听录音，在相应的格子里打“√”。 （6分） 三、Listen and number.听录音，标序号。（9分） pig fox lion cow snake duck

sheep 笔试部分（共80分） 一、Write the questions.将完整的句子写在下面的横线上。(10分) got it Has eyes on a farm it live sheep a it other animals eat it it Is 二、Look and choose.看看它们是谁，将字母填入括号内。(8分) A. B. C. D.

E. F. G. H. ( ) pig ( ) fox ( ) sheep ( ) cat ( ) snake ( ) lion ( ) mouse ( ) elephant 三、Look at the pictures and write the questions.看图片，根据答语写出相应的问题。(10分) No,it doesn’t. Yes,it is.

Yes,it does. Yes,it has. Yes,it does. 四、Choose the right answer.选择正确的答案。(18分) 1、it live on a farm？ 2. it fly？

3. it a cow？ 4. it eat chicken？ 5. you swim？ 6. you all right？ 五、Fill in the numbers.对话排序。(6分) Goodbye. Two apples , please. 45P , please. Thank you.

常用稳压管型号参数对照

常用稳压管型号参数对照 3V到51V 1W稳压管型号对照表1N4727 3V0 1N4728 3V3 1N4729 3V6 1N4730 3V9 1N4731 4V3 1N4732 4V7 1N4733 5V1 1N4734 5V6 1N4735 6V2 1N4736 6V8 1N4737 7V5

1N4739 9V1 1N4740 10V 1N4741 11V 1N4742 12V 1N4743 13V 1N4744 15V 1N4745 16V 1N4746 18V 1N4747 20V 1N4748 22V 1N4749 24V 1N4750 27V 1N4751 30V

1N4753 36V 1N4754 39V 1N4755 43V 1N4756 47V 1N4757 51V 摩托罗拉IN47系列1W稳压管IN4728 3.3v IN4729 3.6v IN4730 3.9v IN4731 4.3 IN4732 4.7 IN4733 5.1

IN4735 6.2 IN4736 6.8 IN4737 7.5 IN4738 8.2 IN4739 9.1 IN4740 10 IN4741 11 IN4742 12 IN4743 13 IN4744 15 IN4745 16 IN4746 18 IN4747 20

IN4749 24 IN4750 27 IN4751 30 IN4752 33 IN4753 34 IN4754 35 IN4755 36 IN4756 47 IN4757 51 摩托罗拉IN52系列 0.5w精密稳压管IN5226 3.3v IN5227 3.6v

(完整版)剑桥小学英语Joinin六年级复习题(二)2-3单元.doc

2011-2012 学年度上学期六年级复习题（Unit2-Unit3 ） 一、听力部分 1、听录音排序 ( ) () ()() () 2、听录音，找出与你所听到的单词属同一类的单词 （） 1. A. spaceman B. pond C . tiger （） 2. A.mascots B. potato C . jeans （） 3. A. door B. behind C . golden （） 4. A. sometimes B. shop C . prince （） 5. A. chair B. who C . sell 3、听录音，将下面人物与他的梦连线 Sarah Tim Juliet Jenny Peter 4、听短文，请在属于Mr. Brown的物品下面打√ ( ) ( ) ( ) ( ) ( ) ( ) ( ) 5、听问句选答句 （） 1. A. Yes, I am B. Yes, I have C . Yes, you do （） 2. A.Pink B. A friendship band C . Yes. （） 3. A. OK B. Bye-bye. C . Thanks, too. （） 4. A. Monday. B. Some juice. C . Kitty. （） 5. A. I ’ve got a shookbag. B. I ’m a student. C . It has got a round face. 6、听短文，选择正确答案 （） 1. Where is Xiaoqing from? She is from . A.Hebei B. Hubei C . Hunan （） 2. She goes to school at . A.7:00 B.7:30 C . 7:15 （） 3. How many classes in the afternoon? classes. A. four B. three C . two （） 4. Where is Xiaoqing at twelve o ’clock? She is . A. at home B. at school C .in the park （） 5. What does she do from seven to half past eight? She . A.watches TV B. reads the book C. does homework

很全的二极管参数

G ENERAL PURPOSE RECTIFIERS – P LASTIC P ASSIVATED J UNCTION 1.0 M1 M2 M3 M4 M5 M6 M7 SMA/DO-214AC G ENERAL PURPOSE RECTIFIERS – G LASS P ASSIVATED J UNCTION S M 1.0 GS1A GS1B GS1D GS1G GS1J GS1K GS1M SMA/DO-214AC 1.0 S1A S1B S1D S1G S1J S1K S1M SMB/DO-214AA 2.0 S2A S2B S2D S2G S2J S2K S2M SMB/DO-214AA 3.0 S3A S3B S3D S3G S3J S3K S3M SMC/DO-214AB F AST RECOVERY RECTIFIERS – P LASTIC P ASSIVATED J UNCTION MERITEK ELECTRONICS CORPORATION

U LTRA FAST RECOVERY RECTIFIERS – G LASS P ASSIVATED J UNCTION

S CHOTTKY B ARRIER R ECTIFIERS

S WITCHING D IODES Power Dissipation Max Avg Rectified Current Peak Reverse Voltage Continuous Reverse Current Forward Voltage Reverse Recovery Time Package Part Number P a (mW) I o (mA) V RRM (V) I R @ V R (V) V F @ I F (mA) t rr (ns) Bulk Reel Outline 200mW 1N4148WS 200 150 100 2500 @ 75 1.0 @ 50 4 5000 SOD-323 1N4448WS 200 150 100 2500 @ 7 5 0.72/1.0 @ 5.0/100 4 5000 SOD-323 BAV16WS 200 250 100 1000 @ 7 5 0.8 6 @ 10 6 5000 SOD-323 BAV19WS 200 250 120 100 @ 100 1.0 @ 100 50 5000 SOD-323 BAV20WS 200 250 200 100 @ 150 1.0 @ 100 50 5000 SOD-323 BAV21WS 200 250 250 100 @ 200 1.0 @ 100 50 5000 SOD-323 MMBD4148W 200 150 100 2500 @ 75 1.0 @ 50 4 3000 SOT-323-1 MMBD4448W 200 150 100 2500 @ 7 5 0.72/1.0 @ 5.0/100 4 3000 SOT-323-1 BAS16W 200 250 100 1000 @ 7 5 0.8 6 @ 10 6 3000 SOT-323-1 BAS19W 200 250 120 100 @ 100 1.0 @ 100 50 3000 SOT-323-1 BAS20W 200 250 200 100 @ 150 1.0 @ 100 50 3000 SOT-323-1 BAS21W 200 250 250 100 @ 200 1.0 @ 100 50 3000 SOT-323-1 BAW56W 200 150 100 2500 @ 75 1.0 @ 50 4 3000 SOT-323-2 BAV70W 200 150 100 2500 @ 75 1.0 @ 50 4 3000 SOT-323-3 BAV99W 200 150 100 2500 @ 75 1.0 @ 50 4 3000 SOT-323-4 BAL99W 200 150 100 2500 @ 75 1.0 @ 50 4 3000 SOT-323- 5 350mW MMBD4148 350 200 100 5000 @ 75 1.0 @ 10 4 3000 SOT-23-1 MMBD4448 350 200 100 5000 @ 75 1.0 @ 10 4 3000 SOT-23-1 BAS16 350 200 100 1000 @ 75 1.0 @ 50 6 3000 SOT-23-1 BAS19 350 200 120 100 @ 120 1.0 @ 100 50 3000 SOT-23-1 BAS20 350 200 200 100 @ 150 1.0 @ 100 50 3000 SOT-23-1 BAS21 350 200 250 100 @ 200 1.0 @ 100 50 3000 SOT-23-1 BAW56 350 200 100 2500 @ 70 1.0 @ 50 4 3000 SOT-23-2 BAV70 350 200 100 5000 @ 70 1.0 @ 50 4 3000 SOT-23-3 BAV99 350 200 100 2500 @ 70 1.0 @ 50 4 3000 SOT-23-4 BAL99 350 200 100 2500 @ 70 1.0 @ 50 4 3000 SOT-23-5 BAV16W 350 200 100 1000 @ 75 0.86 @ 10 6 3000 SOD-123 410-500mW BAV19W 410 200 120 100 @ 100 1.0 @ 100 50 3000 SOD-123 BAV20W 410 200 200 100 @ 150 1.0 @ 100 50 3000 SOD-123 BAV21W 410 200 250 100 @ 200 1.0 @ 100 50 3000 SOD-123 1N4148W 410 150 100 2500 @ 75 1.0 @ 50 4 3000 SOD-123 1N4150W 410 200 50 100 @ 50 0.72/1.0 @ 5.0/100 4 3000 SOD-123 1N4448W 500 150 100 2500 @ 7 5 1.0 @ 200 4 3000 SOD-123 1N4151W 500 150 75 50 @ 50 1.0 @ 10 2 3000 SOD-123 1N914 500 200 100 25 @ 20 1.0 @ 10 4 1000 10000 DO-35 1N4148 500 200 100 25 @ 20 1.0 @ 10 4 1000 10000 DO-35 LL4148 500 150 100 25 @ 20 1.0 @ 10 4 2500 Mini-Melf SOT23-1 SOT23-2 SOT23-3 SOT23-4 SOT23-5 SOT323-1 SOT323-2 SOT323-3 SOT323-4 SOT323-5