Janice A. Espinola

OBJECTIVE: Higher maternal intake of vitamin D during pregnancy is associated with a lower risk of wheezing in offspring. The relationship between cord-blood levels of 25-hydroxyvitamin D (25[OH]D) and childhood wheezing is unknown. We hypothesized that cord-blood levels would be inversely associated with risk of respiratory infection, wheezing, and asthma. PATIENTS AND METHODS: Cord blood from 922 newborns was tested for 25(OH)D. Parents were asked if their child had a history of respiratory infection at 3 months of age or a history of wheezing at 15 months of age and then annually thereafter. Incident asthma was defined as doctor-diagnosed asthma by the time the child was 5 years old and reported inhaler use or wheezing since the age of 4 years. RESULTS: The median cord-blood level of 25(OH)D was 44 nmol/L (interquartile range: 29-78). Follow-up was 89% at the age of 5 years. Adjusting for the season of birth, 25(OH)D had an inverse association with risk of respiratory infection by 3 months of age (odds ratio: 1.00 [reference] for ≥75 nmol/L, 1.39 for 25-74 nmol/L, and 2.16 [95% confidence interval: 1.35-3.46] for <25 nmol/L). Likewise, cord-blood 25(OH)D levels were inversely associated with risk of wheezing by 15 months, 3 years, and 5 years of age (all P < .05). Additional adjustment for more than 12 potential confounders did not materially change these results. In contrast, we found no association between 25(OH)D levels and incident asthma by the age of 5 years. CONCLUSIONS: Cord-blood levels of 25(OH)D had inverse associations with risk of respiratory infection and childhood wheezing but no association with incident asthma.

Importance: Suicide is a leading cause of deaths in the United States. Although the emergency department (ED) is an opportune setting for initiating suicide prevention efforts, ED-initiated suicide prevention interventions remain underdeveloped. Objective: To determine whether an ED-initiated intervention reduces subsequent suicidal behavior. Design, Setting, and Participants: This multicenter study of 8 EDs in the United States enrolled adults with a recent suicide attempt or ideation and was composed of 3 sequential phases: (1) a treatment as usual (TAU) phase from August 2010 to December 2011, (2) a universal screening (screening) phase from September 2011 to December 2012, and (3) a universal screening plus intervention (intervention) phase from July 2012 to November 2013. Interventions: Screening consisted of universal suicide risk screening. The intervention phase consisted of universal screening plus an intervention, which included secondary suicide risk screening by the ED physician, discharge resources, and post-ED telephone calls focused on reducing suicide risk. Main Outcomes and Measures: The primary outcome was suicide attempts (nonfatal and fatal) over the 52-week follow-up period. The proportion and total number of attempts were analyzed. Results: A total of 1376 participants were recruited, including 769 females (55.9%) with a median (interquartile range) age of 37 (26-47) years. A total of 288 participants (20.9%) made at least 1 suicide attempt, and there were 548 total suicide attempts among participants. There were no significant differences in risk reduction between the TAU and screening phases (23% vs 22%, respectively). However, compared with the TAU phase, patients in the intervention phase showed a 5% absolute reduction in suicide attempt risk (23% vs 18%), with a relative risk reduction of 20%. Participants in the intervention phase had 30% fewer total suicide attempts than participants in the TAU phase. Negative binomial regression analysis indicated that the participants in the intervention phase had significantly fewer total suicide attempts than participants in the TAU phase (incidence rate ratio, 0.72; 95% CI, 0.52-1.00; P = .05) but no differences between the TAU and screening phases (incidence rate ratio, 1.00; 95% CI, 0.71-1.41; P = .99). Conclusions and Relevance: Among at-risk patients in the ED, a combination of brief interventions administered both during and after the ED visit decreased post-ED suicidal behavior.

CONTEXT: Vitamin D may be important in the pathogenesis of severe preeclampsia. Given the few effective preventive strategies for severe preeclampsia, studies establishing this link are needed so that effective interventions can be developed. OBJECTIVE: Our objective was to assess whether midgestation vitamin D deficiency is associated with development of severe preeclampsia. DESIGN AND SETTING: We conducted a nested case-control study of pregnant women who had previously given blood for routine genetic multiple marker screening and subsequently delivered at a tertiary hospital between January 2004 and November 2008. PATIENTS: Participants included women with singleton pregnancies in the absence of any chronic medical illnesses. From an overall cohort of 3992 women, 51 cases of severe preeclampsia were matched by race/ethnicity with 204 women delivering at term with uncomplicated pregnancies. Banked maternal serum was used to measure maternal 25-hydroxyvitamin D [25(OH)D]. MAIN OUTCOME MEASURE: The main outcome was severe preeclampsia. RESULTS: Midgestation maternal 25(OH)D concentration was lower in women who subsequently developed severe preeclampsia compared with controls [median (interquartile range), 75 (47-107) nmol/liter vs. 98 (68-113) nmol/liter; P = 0.01]. Midgestation maternal 25(OH)D of less than 50 nmol/liter was associated with an almost 4-fold odds of severe preeclampsia (unadjusted odds ratio, 3.63; 95% confidence interval, 1.52-8.65) compared with midgestation levels of at least 75 nmol/liter. Adjustment for known confounders strengthened the observed association (adjusted odds ratio, 5.41; 95% confidence interval, 2.02-14.52). CONCLUSION: Maternal midgestation vitamin D deficiency was associated with increased risk of severe preeclampsia. Vitamin D deficiency may be a modifiable risk factor for severe preeclampsia.

Juvenile neuronal ceroid lipofuscinosis is caused by mutation of a novel, endosomal/lysosomal membrane protein encoded by CLN3. The observation that the mitochondrial ATPase subunit c protein accumulates in this disease suggests that autophagy, a pathway that regulates mitochondrial turnover, may be disrupted. To test this hypothesis, we examined the autophagic pathway in Cln3Δex7/8 knock-in mice and CbCln3Δex7/8 cerebellar cells, accurate genetic models of juvenile neuronal ceroid lipofuscinosis. In homozygous knock-in mice, we found that the autophagy marker LC3-II was increased, and mammalian target of rapamycin was down-regulated. Moreover, isolated autophagic vacuoles and lysosomes from homozygous knock-in mice were less mature in their ultrastructural morphology than the wild-type organelles, and subunit c accumulated in autophagic vacuoles. Intriguingly, we also observed subunit c accumulation in autophagic vacuoles in normal aging mice. Upon further investigation of the autophagic pathway in homozygous knock-in cerebellar cells, we found that LC3-positive vesicles were altered and overlap of endocytic and lysosomal dyes was reduced when autophagy was stimulated, compared with wildtype cells. Surprisingly, however, stimulation of autophagy did not significantly impact cell survival, but inhibition of autophagy led to cell death. Together these observations suggest that autophagy is disrupted in juvenile neuronal ceroid lipofuscinosis, likely at the level of autophagic vacuolar maturation, and that activation of autophagy may be a prosurvival feedback response in the disease process. Juvenile neuronal ceroid lipofuscinosis is caused by mutation of a novel, endosomal/lysosomal membrane protein encoded by CLN3. The observation that the mitochondrial ATPase subunit c protein accumulates in this disease suggests that autophagy, a pathway that regulates mitochondrial turnover, may be disrupted. To test this hypothesis, we examined the autophagic pathway in Cln3Δex7/8 knock-in mice and CbCln3Δex7/8 cerebellar cells, accurate genetic models of juvenile neuronal ceroid lipofuscinosis. In homozygous knock-in mice, we found that the autophagy marker LC3-II was increased, and mammalian target of rapamycin was down-regulated. Moreover, isolated autophagic vacuoles and lysosomes from homozygous knock-in mice were less mature in their ultrastructural morphology than the wild-type organelles, and subunit c accumulated in autophagic vacuoles. Intriguingly, we also observed subunit c accumulation in autophagic vacuoles in normal aging mice. Upon further investigation of the autophagic pathway in homozygous knock-in cerebellar cells, we found that LC3-positive vesicles were altered and overlap of endocytic and lysosomal dyes was reduced when autophagy was stimulated, compared with wildtype cells. Surprisingly, however, stimulation of autophagy did not significantly impact cell survival, but inhibition of autophagy led to cell death. Together these observations suggest that autophagy is disrupted in juvenile neuronal ceroid lipofuscinosis, likely at the level of autophagic vacuolar maturation, and that activation of autophagy may be a prosurvival feedback response in the disease process. Macroautophagy (herein referred to as autophagy) is a nonselective process by which cytoplasmic constituents are turned over (reviewed in Refs. 1.Cuervo A.M. Trends Cell Biol. 2004; 14: 70-77Abstract Full Text Full Text PDF PubMed Scopus (699) Google Scholar and 2.Yorimitsu T. Klionsky D.J. Cell Death Differ. 2005; 12: 1542-1552Crossref PubMed Scopus (1169) Google Scholar). Elegant work in yeast has led to the genetic dissection of many of the proteins involved in this pathway (3.Tsukada M. Ohsumi Y. FEBS Lett. 1993; 333: 169-174Crossref PubMed Scopus (1357) Google Scholar, 4.Thumm M. Egner R. Koch B. Schlumpberger M. Straub M. Veenhuis M. Wolf D.H. FEBS Lett. 1994; 349: 275-280Crossref PubMed Scopus (476) Google Scholar), which are mostly conserved in mammals (5.Mizushima N. Ohsumi Y. Yoshimori T. Cell Struct. Funct. 2002; 27: 421-429Crossref PubMed Scopus (741) Google Scholar, 6.Levine B. Klionsky D.J. Dev. Cell. 2004; 6: 463-477Abstract Full Text Full Text PDF PubMed Scopus (3103) Google Scholar). The pathway is initiated at times of stress or nutritional deprivation to generate metabolites for cellular survival, but autophagy is also responsible for normal housekeeping. Although the regulatory control of autophagy remains to be fully elucidated, the general process from engulfment to degradation has been delineated (2.Yorimitsu T. Klionsky D.J. Cell Death Differ. 2005; 12: 1542-1552Crossref PubMed Scopus (1169) Google Scholar). Initiation of the pathway involves de novo formation of an isolation membrane that expands to engulf cytoplasmic constituents, including organelles, and encloses to form a double membrane autophagic vacuole. This formed autophagic vacuole is then increasingly acidified, and proteases are delivered through fusion with late endosomes and lysosomes. The inner membrane is degraded to form a single membrane autolysosome, where constituents are finally completely degraded for recycling of metabolites to the cell. Autophagy is emerging as a major pathway involved in a number of neurodegenerative diseases, including Parkinson disease (7.Anglade P. Vyas S. Javoy-Agid F. Herrero M.T. Michel P.P. Marquez J. Mouatt-Prigent A. Ruberg M. Hirsch E.C. Agid Y. Histol. Histopathol. 1997; 12: 25-31PubMed Google Scholar, 8.Stefanis L. Larsen K.E. Rideout H.J. Sulzer D. Greene L.A. J. Neurosci. 2001; 21: 9549-9560Crossref PubMed Google Scholar), Huntington disease (9.Kegel K.B. Kim M. Sapp E. McIntyre C. 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In each case, autophagic vacuoles accumulate, suggesting that activation of autophagy is a common feature of neurodegeneration. However, the events leading to autophagic activation and how these impact any given disease process is not yet understood, and it is likely that autophagic involvement will differ in each case, since the diseases are clinically and pathologically distinct. The neuronal ceroid lipofuscinoses (NCLs) 3The abbreviations used are: NCL, neuronal ceroid lipofuscinosis; 3-MA, 3-methyladenine; AV, autophagic vacuole(s); AV15, light autophagic vacuolar fraction; AV20, heavy autophagic vacuolar fraction; E, tubulovesicular fraction; ER, endoplasmic reticulum; JNCL, juvenile neuronal ceroid lipofuscinosis; mTOR, mammalian target of rapamycin; Tricine, N-[2-hydroxy-1, 1-bis(hydroxymethyl)ethyl]glycine; Bistris, 2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)propane-1,3-diol.3The abbreviations used are: NCL, neuronal ceroid lipofuscinosis; 3-MA, 3-methyladenine; AV, autophagic vacuole(s); AV15, light autophagic vacuolar fraction; AV20, heavy autophagic vacuolar fraction; E, tubulovesicular fraction; ER, endoplasmic reticulum; JNCL, juvenile neuronal ceroid lipofuscinosis; mTOR, mammalian target of rapamycin; Tricine, N-[2-hydroxy-1, 1-bis(hydroxymethyl)ethyl]glycine; Bistris, 2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)propane-1,3-diol. are the most common cause of neurodegeneration among children. These recessively inherited disorders typically manifest with blindness, seizures, progressive cognitive and motor failure, and the NCLs are always fatal (14.Wisniewski K.E. Neurology. 2001; 57: 576-581Crossref PubMed Scopus (91) Google Scholar). Although heterogeneous in genetic origin and some disease features, including age at onset, nearly all of the NCLs lead to lysosomal accumulation of subunit c of the mitochondrial ATPase F0 complex (15.Palmer D.N. Fearnley I.M. Walker J.E. Hall N.A. M. J. Genet. PubMed Scopus Google Scholar), in the of this protein the autophagic genetic cause with subunit c accumulation (reviewed in 2004; 14: PubMed Scopus Google Scholar). Juvenile the most common form of NCL, from of Full Text PDF PubMed Scopus Google Scholar). major mutation in the for the of disease Full Text PDF PubMed Scopus Google Scholar). This mutation and and the to protein which are Full Text PDF PubMed Scopus Google Scholar, L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google Scholar). The and is a protein that to the late endosomal/lysosomal where it has been in and T. B. F. Nat. Genet. PubMed Scopus Google Scholar, D. Hum. Mol. Genet. 2005; 14: PubMed Scopus Google and K. K. L. A. A. Hum. Mol. Genet. 2004; PubMed Scopus Google Scholar, E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). knock-in L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google and cerebellar models E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google that the common mutation in the in JNCL, and in homozygous Cln3Δex7/8 mice progressive accumulation of subunit c that L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google Scholar), and homozygous CbCln3Δex7/8 cerebellar subunit c when by aging at E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). In the we test the that autophagy is involved in the disease process and the subunit c accumulation of These of autophagy in and a for the in normal used in these were as and a from J. M. M. Y. R. K. Uchiyama Y. T. E. J. PubMed Scopus Google and c E. J. D. K. T. J. Biochem. PubMed Scopus Google from and and target of rapamycin and were as and from rapamycin and from Cln3Δex7/8 and and wildtype were and as L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google Scholar). and of vacuoles were isolated from by in an isolation from L. J. J. Cell Biol. PubMed Scopus Google Scholar), as W.H. Cuervo A.M. Kumar A. Peterhoff C.M. Schmidt S.D. Lee J.H. Mohan P.S. Mercken M. Farmery M.R. Tjernberg L.O. Jiang Y. Duff K. Uchiyama Y. Naslund J. Mathews P.M. Cataldo A.M. Nixon R.A. J. Cell Biol. 2005; 171: 87-98Crossref PubMed Scopus (797) Google Scholar). from were by for the for each was as E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). The of the and the protein in the and each were for of the protein in the and of ultrastructural an of the was in and in with for of were for as L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google Scholar). of each were for subunit c and which were of each were the to the were by by the over times from to were a and The of was with the times were and were to used for further were from the subunit c in and in a R. Fearnley I.M. D.N. Walker J.E. J. Biol. 2004; Full Text Full Text PDF PubMed Scopus Google the from was used for of subunit in the subunit c is as the of and subunit from to the protein in the to from were at a of a at this was were for the number of vesicles of the double and membrane were as vesicles with a double were as vesicles of heterogeneous with or vesicles of a double were as heterogeneous vesicles of membrane a single or double were as vesicles heterogeneous including a single each an test was to the for wildtype homozygous Cln3Δex7/8 were significantly and Cell and cell were Full Text PDF PubMed Scopus Google and were for these as In 20: PubMed Scopus Google Scholar). CbCln3Δex7/8 cerebellar cell also been E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). CbCln3Δex7/8 cerebellar were with the to the were in or at the and to to the The was for with to or The and cell were as E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). endocytic was the of or and times were from to was a or a and were and were always with for wild-type and homozygous CbCln3Δex7/8 cells. of in the was used as of the was for for each was a cell the were of cell at the compared with the for each cell was a of in cell at the was an c and homozygous CbCln3Δex7/8 cerebellar were at a of and to This for the of the The was for or were in and for subunit c as E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). c were in with a was an of Autophagy in Cln3Δex7/8 test the that autophagy is involved in the of JNCL, we examined the of autophagy in Cln3Δex7/8 knock-in mice. is a protein that is activation of autophagy LC3-II with the and membrane and is a marker of autophagy N. Int. J. Biochem. Cell Biol. 2004; 36: PubMed Scopus Google Scholar). with an that of were observed in from control mice or Cln3Δex7/8 were observed in homozygous Cln3Δex7/8 in to mitochondrial ATPase subunit c including the of the cell of the and L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google To the of that was in the LC3-II which than and is the form with the we and of and LC3-II in wild-type and homozygous Cln3Δex7/8 in homozygous Cln3Δex7/8 knock-in to wild-type were observed at each age These observations are with an activation of autophagy in homozygous Cln3Δex7/8 mice. in the autophagic we a W.H. Cuervo A.M. Kumar A. Peterhoff C.M. Schmidt S.D. Lee J.H. Mohan P.S. Mercken M. Farmery M.R. Tjernberg L.O. Jiang Y. Duff K. Uchiyama Y. Naslund J. Mathews P.M. Cataldo A.M. Nixon R.A. J. Cell Biol. 2005; 171: 87-98Crossref PubMed Scopus (797) Google to autophagic vacuoles from wildtype and Cln3Δex7/8 knock-in for and This the isolation of autophagic vacuoles of a light in and a heavy of the of autophagy in and the observed in in homozygous Cln3Δex7/8 mice L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google Scholar), we examined the To the was in the which suggest a for in the autophagic we of wild-type and homozygous Cln3Δex7/8 a J. M. M. Y. R. K. Uchiyama Y. T. E. J. PubMed Scopus Google Scholar). wild-type we observed a in in the and tubulovesicular mostly of which was in the AV15, AV20, and lysosomal and this was homozygous Cln3Δex7/8 as with and in homozygous Cln3Δex7/8 L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google Scholar), a at with and lysosomes was homozygous Cln3Δex7/8 in mostly with the and a at with J. M. M. Y. R. K. Uchiyama Y. T. E. J. PubMed Scopus Google Scholar), were not wildtype and homozygous Cln3Δex7/8 of from Cln3Δex7/8 the of from wild-type and homozygous Cln3Δex7/8 we an of reduced protein in the AV15, AV20, and lysosomal from homozygous Cln3Δex7/8 compared with the wild-type which with age not These observations in the and lysosomal from homozygous Cln3Δex7/8 we to further the ultrastructural morphology of the and lysosomal of the AV15, AV20, and lysosomal from wildtype mice an progressive and degradation of with where the were in the vesicles in the lysosomal with observations (12.Yu W.H. Kumar A. Peterhoff C. Shapiro Kulnane L. Uchiyama Y. Lamb B.T. Cuervo A.M. Nixon R.A. Int. J. Biochem. Cell Biol. 2004; 36: 2531-2540Crossref PubMed Scopus (243) Google Scholar, 13.Yu W.H. Cuervo A.M. Kumar A. Peterhoff C.M. Schmidt S.D. Lee J.H. Mohan P.S. Mercken M. Farmery M.R. Tjernberg L.O. Jiang Y. Duff K. Uchiyama Y. Naslund J. Mathews P.M. Cataldo A.M. Nixon R.A. J. Cell Biol. 2005; 171: 87-98Crossref PubMed Scopus (797) Google Scholar). In this was not observed in the from homozygous Cln3Δex7/8 vesicles in and in the lysosomal from homozygous Cln3Δex7/8 were of in with the observed in Moreover, with were in the from wild-type mice, but these were not observed in the homozygous Cln3Δex7/8 c in in Cln3Δex7/8 and of mitochondrial subunit c in is to at the level of the (15.Palmer D.N. Fearnley I.M. Walker J.E. Hall N.A. M. J. Genet. PubMed Scopus Google Scholar). However, given that the subunit c protein is to to the the autophagic pathway 2002; PubMed Scopus Google Scholar), we to the in which subunit c the and lysosomal from wild-type and homozygous Cln3Δex7/8 knock-in we a of subunit c by and of subunit c of this protein in mitochondrial Moreover, with of subunit c protein was also in and lysosomes from wild-type and homozygous Cln3Δex7/8 knock-in of subunit by of to protein in each a of subunit c in the homozygous Cln3Δex7/8 knock-in to wild-type The homozygous Cln3Δex7/8 knock-in lysosomal also subunit c to the wild-type lysosomal Intriguingly, the from Cln3Δex7/8 did not subunit c major in subunit c were observed in the tubulovesicular and mitochondrial marker the of the and major in of these were observed the wild-type and homozygous Cln3Δex7/8 However, LC3-II was in the from homozygous Cln3Δex7/8 compared with with observations in and were also in the from homozygous Cln3Δex7/8 compared with and not the progressive of subunit c in homozygous Cln3Δex7/8 mice and L.A. Lee A.M. K. Hum. Mol. Genet. 2002; PubMed Google Scholar, M. J. M. 1997; PubMed Scopus Google Scholar), the accumulation of subunit c in the was at subunit c from and lysosomal in homozygous Cln3Δex7/8 knock-in from to Surprisingly, however, an in subunit c from and lysosomal was also observed for wild-type the subunit c accumulation in homozygous Cln3Δex7/8 knock-in which in the subunit c was in the from wild-type Moreover, subunit c in wild-type and homozygous Cln3Δex7/8 knock-in and lysosomal was of in observations at the level of suggest that autophagy may be in and from homozygous Cln3Δex7/8 knock-in mice, the of the formed vesicles may be The pathway is to autophagy T. Ohsumi Y. J. Biol. Full Text Full Text PDF PubMed Scopus Google Scholar, J. Mol. Cell. PubMed Scopus Google Scholar). To the involvement of the pathway in we the of activation in homozygous Cln3Δex7/8 knock-in mice. in we found significantly reduced of in homozygous Cln3Δex7/8 compared with Cln3Δex7/8 and wild-type To observations in homozygous Cln3Δex7/8 mice were also in cell we the of activation to and CbCln3Δex7/8 cerebellar cells, which we from Cln3Δex7/8 knock-in mice E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). in the of the target of mTOR, were significantly reduced to observed in control cell and in wild-type cerebellar cell and was also not in CbCln3Δex7/8 autophagy may be in genetic models of JNCL, the formed may not mature leading to a in This a of autophagic in CbCln3Δex7/8 cerebellar cells. that homozygous CbCln3Δex7/8 not subunit c accumulation normal subunit c were by aging at which may autophagy E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google Scholar). To test stimulation of autophagy to subunit c accumulation in homozygous CbCln3Δex7/8 cerebellar cells, we with the and subunit c The of rapamycin to autophagy was in CbCln3Δex7/8 cell by LC3-II with in observations homozygous CbCln3Δex7/8 cerebellar cell to than wild-type cells, and rapamycin the in wild-type and homozygous CbCln3Δex7/8 stimulation of autophagy by rapamycin led to subunit c formation in homozygous CbCln3Δex7/8 cerebellar further that subunit c accumulation in the autophagic also examined by with a In wild-type cerebellar cells, we observed and vesicles that were in with S. C. T. E. P. J. Cell 2004; PubMed Scopus Google However, homozygous CbCln3Δex7/8 cerebellar the but with a in fusion with late endosomes and which to and activation for degradation of the (5.Mizushima N. Ohsumi Y. Yoshimori T. Cell Struct. Funct. 2002; 27: 421-429Crossref PubMed Scopus (741) Google Scholar, 6.Levine B. Klionsky D.J. Dev. 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Neurosci. 2004; PubMed Scopus Google Scholar), the endocytic and lysosomes and dyes were reduced in homozygous CbCln3Δex7/8 cerebellar cells, to wild-type cerebellar Upon rapamycin in wild-type and homozygous CbCln3Δex7/8 cells, the endocytic and were increased, and overlap of the dyes was of the homozygous CbCln3Δex7/8 cerebellar with the wild-type a in to that observed for cells, but we also observed in the of these in homozygous CbCln3Δex7/8 cells, to the wild-type These observations the that endocytic and which involves is in homozygous CbCln3Δex7/8 cerebellar cells. of Autophagy Cell is a prosurvival pathway and an pathway leading to cell N. Cell Death Differ. 2005; 12: PubMed Scopus Google Scholar). To the of autophagy and cell in wild-type and homozygous CbCln3Δex7/8 cells, we cell with rapamycin or 3-MA, a as an of autophagy W.H. Cuervo A.M. Kumar A. Peterhoff C.M. Schmidt S.D. Lee J.H. Mohan P.S. Mercken M. Farmery M.R. Tjernberg L.O. Jiang Y. Duff K. Uchiyama Y. Naslund J. 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Biol. 2005; PubMed Scopus Google Scholar), in cell in wild-type and homozygous CbCln3Δex7/8 cells, but in the of homozygous CbCln3Δex7/8 was significantly compared with wild-type cerebellar cell at all with control and these where cell was observed not These and accurate and cerebellar cell which the common an for autophagy in the of ATPase subunit c and in disease of ATPase subunit c in membrane and in aging were of autophagy in homozygous for the common including cells, as in the in The observation that subunit c accumulation in homozygous Cln3Δex7/8 is not to the but is most in the light suggests that is a in the of to fully in The ultrastructural in the degradation of the this Although the of the are not yet understood, the light with the is with and J. Biol. 20: Google Scholar). the of the in this this the observations of accumulation of subunit c in the light are with a in of in of the autophagic has been in as where autophagic through to the is to be (12.Yu W.H. Kumar A. Peterhoff C. Shapiro Kulnane L. Uchiyama Y. Lamb B.T. Cuervo A.M. Nixon R.A. Int. J. Biochem. Cell Biol. 2004; 36: 2531-2540Crossref PubMed Scopus (243) Google Scholar, 13.Yu W.H. Cuervo A.M. Kumar A. Peterhoff C.M. Schmidt S.D. Lee J.H. Mohan P.S. Mercken M. Farmery M.R. Tjernberg L.O. Jiang Y. Duff K. Uchiyama Y. Naslund J. Mathews P.M. Cataldo A.M. Nixon R.A. J. Cell Biol. 2005; 171: 87-98Crossref PubMed Scopus (797) Google Scholar). The that to the accumulation in may fusion the formed and late endosomes and leading to a in and subunit which in the This is by cell in homozygous CbCln3Δex7/8 cerebellar in which and lysosomal is Moreover, the observation that is in the of is for the that a in the The that to of the or it may be that membrane protein or by as membrane with these and lysosomal K. K. L. A. A. Hum. Mol. Genet. 2004; PubMed Scopus Google Scholar, E. Wolf P. T. A.M. D. E. Neurosci. 2004; PubMed Scopus Google and lysosomal T. B. F. Nat. Genet. 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