text:
Type: Bug
Status: Closed
Priority: Major
Resolution: Fixed
Affects Version/s: 1.9.2
Fix Version/s: 1.9.10
Component/s: Backup
Environment:Moodle 1.9.2+ (Build: 20080806)
Apache/2.2.9 (Win32) mod_ssl/2.2.9 OpenSSL/0.9.8h PHP/5.2.6
Windows 2003 SP1
SQL 2005 (via FreeTDS)
Database:Microsoft SQL
Affected Branches:MOODLE_19_STABLE
Fixed Branches:MOODLE_19_STABLE
When viewing Backup reports in Moodle, you receive the following error:
Ambiguous column name 'laststarttime'.
SELECT DISTINCT laststarttime,laststarttime FROM mdl_backup_log WHERE courseid = '16' ORDER BY laststarttime DESC
? line 686 of lib\dmllib.php: call to debugging()
? line 966 of lib\dmllib.php: call to get_recordset_sql()
? line 102 of admin\report\backups\index.php: call to get_records_sql()
No logs have been found
The fix simply appears to be removing the extraneous 'laststarttime' from the SQL query on line 99 of admin\report\backups\index.php.
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I have enabled the automated backups function in Moodle 1.9.9+ (Build: 20100908) and they run nicely, except for a few "Error" messages that I see in the Last execution log of the Backup report. This implies that there is something wrong with the backup files that Moodle is creating. I was obviously eager to determine what these errors are so I tried view the Detailed execution log for the courses that are showing the "Error" message, but the only message I ever get is "No logs have been found".
Eventually I turned the Debug messages option up to the maximum level (Developer) and tried to view the Detailed execution logs for the problem courses again. The debug text shows the following message:
Ambiguous column name 'laststarttime'.
SELECT DISTINCT laststarttime,laststarttime FROM mdl_backup_log WHERE courseid = '1' ORDER BY laststarttime DESC
line 686 of lib\dmllib.php: call to debugging()
line 966 of lib\dmllib.php: call to get_recordset_sql()
line 99 of admin\report\backups\index.php: call to get_records_sql()
I then went into \admin\report\backups\index.php and changed line 96 from:
$executions = get_records_sql("SELECT DISTINCT laststarttime,laststarttime
FROM {$CFG->prefix}backup_log
WHERE courseid = '$courseid'
ORDER BY laststarttime DESC");
$executions = get_records_sql("SELECT DISTINCT laststarttime
FROM {$CFG->prefix}backup_log
WHERE courseid = '$courseid'
ORDER BY laststarttime DESC");
In other words, I removed one of the "laststarttime" columns from the sql statement. Now when I view the Detailed execution logs for any of my courses I see the whole log instead of a "No logs have been found" message.
I'm happy to make this change on my production box so that I can troubleshoot the problem backups, but will this change cause any regressions?
I see that the admin\report\backups\index.php file in Moodle 2.0 Preview 4+ (Build: 20100902) (2010082600) the code has changed to:
$executions = $DB->get_records_sql("SELECT DISTINCT laststarttime,laststarttime
WHERE courseid = ? AND backuptype = ?
ORDER BY laststarttime DESC", array($courseid,'scheduledbackup'));
but it still selects the laststarttime column twice.
SELECT DISTINCT laststarttime,laststarttime FROM...
valid or should it be
SELECT DISTINCT laststarttime FROM...?
Change applied to CVS, that was one "old trick" needed in previous versions to be able to get only one field, but in 1.9.something we fixed the limit and all the occurrences (usually "id, id"). It seems that we forgot this ("laststarttime, laststarttime").
Also have fixed 2.0 although that report surely will change because logs aren't stored there anymore.
Thanks and ciao
Eloy, not sure if I should raise this here on in the forums - but why would I see this error now when all I have done is move a fully functioning Moodle site onto a new webserver? The DB server has remained the same.
Running exactly the same moodle version?
Somewhere in the 1.9.x life the change happened so, for example, if your original server was running 1.9.5, then it could be accepting the double "laststarttime", but if the new server is 1.9.7, then it has stopped accepting the doubles.
(note the versions above, 1.9.5 and 1.9.7, are invented, I haven't looked exactly when the change happened)
Could that be your case?
Hi Eloy,
It's exactly the same Moodle version. We just moved the code from one server to the next.
wow, and in the old one you're getting the logs displayed and in the other you get the error?
same code-base, same DB server, same... everything? Crazy! I cannot imagine any cause for that. Sorry.


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Corticospinal motor neurons (CSMN) receive, integrate, and relay cerebral cortex's input toward spinal targets to initiate and modulate voluntary movement. CSMN degeneration is central for numerous motor neuron disorders and neurodegenerative diseases. Previously, 5 patients with mutations in the ubiquitin carboxy-terminal hydrolase-L1 (UCHL1) gene were reported to have neurodegeneration and motor neuron dysfunction with upper motor neuron involvement. To investigate the role of UCHL1 on CSMN health and stability, we used both in vivo and in vitro approaches, and took advantage of the Uchl1nm3419 (UCHL1−/−) mice, which lack all UCHL1 function. We report a unique role of UCHL1 in maintaining CSMN viability and cellular integrity. CSMN show early, selective, progressive, and profound cell loss in the absence of UCHL1. CSMN degeneration, evident even at pre-symptomatic stages by disintegration of the apical dendrite and spine loss, is mediated via increased ER stress. These findings bring a novel understanding to the basis of CSMN vulnerability, and suggest UCHL1−/− mice as a tool to study CSMN pathology.
Corticospinal motor neurons (CSMN) are exceptional in their ability to receive, integrate, translate, and transmit information from the cerebral cortex to spinal cord targets (Molyneaux, Arlotta, Macklis 2007; Molyneaux Arlotta, Menezes et al. 2007; Greig et al. 2013). They are the cortical component of motor neuron circuitry that initiates and modulates very precise aspects of voluntary movement (Alstermark and Isa 2012). Execution of such complicated tasks emphasizes the importance of corticospinal motor neuron (CSMN) health and stability for proper motor neuron circuitry. Their degeneration, therefore, has significant impact on various neurodegenerative diseases in which voluntary movement is impaired, such as amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD; Novarino et al. 2014).
Identification of mutations in the ubiquitin carboxy-terminal hydrolase-L1 (UCHL1) gene of 2 patients with a PD-like phenotype (Leroy et al. 1998), and a recent clinical report describing 3 siblings with progressive and early neurodegeneration, and upper motor neuron dysfunction (Bilguvar et al. 2013) sparked interest in the potential importance of UCHL1 function in a spectrum of neurodegenerative diseases, especially those that affect voluntary movement and motor neuron circuitry.
UCHL1 is a unique deubiquitinating enzyme that has both hydrolase and ligase activities: It can remove ubiquitin from poly-ubiquitin chains (Osaka et al. 2003), and also add ubiquitin to already ubiquitinylated proteins (Liu et al. 2002). Since inhibition of UCHL1 results in a 50% reduction of free ubiquitin in vitro (Leroy et al. 1998; Cartier et al. 2009), UCHL1 was suggested to be critically important for maintaining free ubiquitin levels and for the proper function of the ubiquitin–proteasome system in neurons (Day and Thompson 2010). Abnormalities in protein homeostasis have been implicated in motor neuron diseases (Nishimura et al. 2004; Petrucelli and Dawson 2004; Johnson et al. 2010; Deng et al. 2011; Rubino et al. 2012), but the impact of a dysfunctional UCHL1 on the health and stability of motor neurons, especially upper motor neurons, remains poorly understood.
To reveal the potential link between UCHL1 function and especially the cortical component of the motor neuron circuitry, we investigated CSMN health in Uchl1nm3419 (UCHL1−/−) mice, which lack all UCHL1 function (Walters et al. 2008), and display motor function defects. Using retrograde labeling, molecular marker expression analysis, and adeno-associated virus (AAV)-mediated anterograde and retrograde transduction, we demonstrate early, selective, progressive, and profound CSMN degeneration in the absence of UCHL1 function. Other projection neurons in the motor cortex, such as callosal projection neurons (CPN), are not affected. Vulnerable CSMN display cellular abnormalities with vacuolated apical dendrites and spine loss, as early as postnatal day (P) 60, and their degeneration is linked to increased ER stress both in vivo and in vitro. Our findings begin to reveal a unique importance of UCHL1 function for CSMN health, and suggest that UCHL1−/− mice can be used as a tool to study the underlying mechanisms for CSMN dysfunction in diseases with prominent upper motor neuron involvement.
Materials and Methods
All animal procedures were approved by Northwestern University and the Massachusetts General Hospital–Harvard Medical School Animal Care and Use Committee, and conformed to the standards of the National Institutes of Health. Uchl1nm3419 (UCHL1−/−) mice carry a spontaneous 795-bp intragenic deletion that results in the removal of 24 bp of exon 6 and 771 bp of intron 6 (Fig. 1A; Walters et al. 2008). Heterozygous mice (UCHL1+/−) were viable, fertile, and bred to generate UCHL1-deficient (UCHL1−/−) mice. All mice were on a C57BL/6J background. Survival times and motor function defects were comparable between males and females with 100% penetrance. Primers used to determine genotypes are UCHL1 forward: tggacggctgtgtgtgctaatg, wild-type (WT) reverse: ctaagggaagggtcttgctcatc, mutant (Mt) reverse: gtcatctacctgaagagagccaag, yielding 668 bp WT and 334 bp Mt PCR products, respectively.
Postmortem Human Brain Samples
Acquisition of postmortem human tissue was carried out according to protocols approved by an institutional review board, and clinical records were available for every subject. Two control (A12–105, 52-year-old male and A12–106, 58-year-old female) cases were made available by the University of Chicago, IL, USA. A neurologist examined all human subjects and a neuropathologist reviewed autopsy tissue. Control case A12–105: 52-year-old male with a history of coronary artery disease, heart failure, morbid obesity, and implanted defibrillator who died of cardiac decompensation. Control case A12–106: 58-year-old female with pulmonary hypertension, scleroderma, and Raynaud's disease who died of cardiorespiratory failure. Brains were fixed in 20% neutral-buffered formalin for 1 week, and sections were paraffin-embedded. Areas of the primary motor cortex were retrieved; 4-μm thick serial sections were cut and used for immunocytochemical analyses.
Rotarod Analysis, DigiGait, and Grip Test
Fore and hind limbs of WT and UCHL1−/− mice were painted with red and black paints, respectively, prior to walking on white paper in an enclosed area. A rotating rod that accelerates linearly from 4 to 40 rpm (Rotarod, Ugo Basile) was used and an average time spent on the rotating rod for 3 consecutive trials was calculated for maximum 5 min. Gait analysis was performed as previously described (Gogliotti et al. 2011) using an enclosed DigiGait Imaging system (Mouse Specifics, Quincy, MA, USA). A transparent treadmill was adjusted to 17 cm/s. Swing and stride parameters were determined from a 5-s video. Time each mouse spent hanging on to an upside-down wire mesh (50 cm above a bench) was recorded in 3 consecutive trials, for Grip testing.
Generation of AAV
AAV vectors were generated by the University of Pennsylvania Vector Core facility by triple transfection of subconfluent HEK293 cells using 3 plasmids: an AAV trans-plasmid encoding AAV2 capsid, an adenovirus helper plasmid pΔF6, and an AAV cis-shuttle plasmid expressing eGFP driven by a CMV promoter (pENN.AAV.CMV.PI.eGFP .WPRE.bGH). The culture medium was collected, concentrated by tangential flow filtration, and purified by iodixanol gradient ultracentrifugation as previously described (Lock et al. 2010).
Anterograde and Retrograde Labeling and Transduction Surgeries
Surgeries were performed on a stereotaxic platform. Microinjections were performed using pulled-beveled glass micropipettes attached to a nanojector (Drummond Scientific, Broomall, PA, USA).
CSMN Retrograde Labeling and Transduction
CSMN were retrogradely labeled either by injection of fluorescent microspheres (LumaFluor, Inc., Naples, FL, USA; ∼97 nL for P2, and ∼207 nL for P38 injections), or by 0.5% Fluoro-Gold (FG; Fluorochrome LLC, Denver, CO, USA; 96 nL in 0.9% saline solution) at P30 (Ozdinler et al. 2011; Jara et al. 2012). Mice were sacrificed either at P40, P65, P80, or P100 (n = 3). To study CSMN cytoarchitecture, AAV-encoding eGFP (AAV2-eGFP; 621 nL containing 1.16 × 109 viral particles) was injected into the CST and CSMN were retrogradely transduced (n = 3) as described (Jara et al. 2012).
CSMN Anterograde Labeling
CSMN were anterogradely labeled by AAV-eGFP injection into the motor cortex. A unilateral craniotomy of approximately 5 mm2 (coordinates = +0.5 mm anterior-posterior; 1.5 mm mediolateral) was performed into the left hemisphere using a microdrill (Fine Science Tools, Foster City, CA, USA). Viral particles (1.16 × 109) were injected (10 injections, total 621 nL/mouse). Surgeries were performed at P30, and mice were sacrificed at P60 (n = 3).
CPN Retrograde Labeling
To retrogradely label CPN in the contralateral hemisphere, a craniotomy of approximately 5 mm2 (coordinates = +0.5 mm anterior-posterior; 1.5 mm mediolateral) was performed into the left hemisphere using microdrill (Fine Science Tools, Foster City, CA, USA). About 64 nL of 0.5% FG in 0.9% saline solution was injected (10 injections/mouse). Surgeries were performed at P30, and mice were sacrificed at P100 (n = 3).
Tissue Collection and Histology
Mice were deeply anesthetized with ketamine (90 mg/kg) and xylazine (10 mg/kg), and perfused with 4% PFA in PBS. The brain was removed intact from each mouse, post-fixed by 4% PFA overnight, and kept in PBS–sodium azide (0.01%) at 4 °C. Brains were sectioned (coronal; 50 μm) using a vibrating microtome (VT1000S, Leica Instruments, Nussloch, Germany). Nissl and hematoxylin–eosin (H&E) staining were performed. Optic nerves were embedded in resin and semi-thin sections were stained with toluidine blue.
Western Blot
Brains and spinal cords were isolated from WT and UCHL1−/− mice at P40 and P100. Protein lysates from primary motor cortex as well as from cervical and lumbar spinal cord were obtained, resolved by 10% SDS–PAGE, and transferred onto a PVDF membrane (Bio-Rad, Hercules, CA, USA) as previously described (Jara et al. 2007). Blots were incubated with primary antibodies overnight at 4 °C and developed by enhanced chemiluminescence (Millipore, Temecula, CA, USA) using HRP-conjugated secondary antibodies.
Immunocytochemistry and In Situ Hybridization
Immunocytochemistry was performed on every 12th coronal section of mouse brains. Antigen retrieval was performed for Ctip2, PERK, and PDI immunocytochemistry; sections were treated with 0.01 M sodium citrate, pH 9.0, at 80 °C water bath for 3 h prior to incubation with primary antibody. After PBS washes, either fluorescent-conjugated (Alexa 488, Cy3) or biotinylated secondary antibodies (Vector Laboratories, Burlingame, CA, USA) were used.
For ER stress analysis on CSMN, PERK and PDI expression were studied on neurons that express Ctip2 (developed with alkaline phosphatase-conjugated secondary antibody and AP substrate Kit II; Vector Laboratories, Inc., Burlingame, CA, USA). Biotinylated secondary antibodies, ABC Kit (Vector Laboratories, Inc.), and NovaRED substrate (Vector Laboratories, Inc.) were used to determine PDI and PERK expression, as directed by the manufacturer.
Primary antibodies were: anti-Ctip2 (1 : 500; Abcam); anti-FG (1 : 1000; Fluorochrome, LLC); anti-GFP (1 : 1000; Invitrogen); anti-Lmo4 (1 : 500; Millipore); anti-NeuN (1 : 1000; Millipore); anti-PERK (1 : 1000; Cell Signaling Technology); anti-PDI (1 : 1000; Cell Signaling Technology); anti-ubiquitinylated proteins (1 : 1000; clone FK1; Millipore), and anti-UCHL1 (1 : 1000; ProteinTech).
For in situ hybridization, Crym and Fezf2 cRNAs were digoxigenin-labeled, and all probes were alkali hydrolyzed to an average length of 250–500 bp. Serial frozen sections (16 μm) were collected on slides, post-fixed with 4% PFA, rinsed in PBS, submerged in hybridization buffer containing a probe (100 ng/mL), and hybridized overnight at 68 °C. Sense probes were used as negative controls in all experiments and in situ hybridization was performed as previously described (Arlotta et al. 2005).
Tissue Culture
P2 motor cortices were dissected, dissociated, and cultured on glass coverslips (4.44 × 104cells per 18 mm diameter coverslip, Fisherbrand) coated with poly-l-lysine (10 mg/mL, Sigma). Neurons were cultured in serum-free medium [SFM; 0.034 mg/L BSA, 1 mM l-glutamine, 25 U/mL penicillin, 0.025 mg/mL streptomycin, 35 mM glucose, and 0.5% B27 in Neurobasal-A medium (Life Technologies)] in a humidified tissue culture incubator in the presence of 5% CO2 at 37 °C.
To induce ER stress, Tunicamycin (Sigma-Aldrich; 100 μM) or proteasome inhibitor Bortezomib (Santa Cruz Biotechnology; 100 nM) was added directly into culture medium after 1 day in vitro (DIV). Guanabenz (Sigma-Aldrich; 100 μM) was added at 1DIV to reduce ER stress. Dissociated cortical neurons were fixed prior to treatment, after 3 h and after 10 h of treatment to assess the levels of PERK and PDI and the changes in survival of CSMN and other cortical neurons. Percent survival was determined upon application of ethidium homodimer (EtHom; Invitrogen; 2 μM), the dead cell indicator, for 30 min prior to fixation (2% PFA, 15 min RT).
CSMN were identified based on their Ctip2 expression and large soma size. Ctip2/PERK and Ctip2/PDI co-expression analyses were performed in a serial staining protocol in which cells were stained with PERK or PDI antibody first, using an alkaline phosphatase-conjugated secondary antibody and AP Substrate Kit III (Vector Laboratories, Inc.), followed by Ctip2 immunocytochemistry using a biotinylated secondary antibody and ABC kit with NovaRED substrate (Vector Laboratories, Inc.). To determine percent CSMN survival, Ctip2 immunocytochemistry was performed on cultures treated with EtHom.
System Analysis
Proteins that are reported to have direct interactions with UCHL1 were determined using curated information generated from published data sets [Ingenuity Pathway Analysis (IPA); QIAGEN Comp, LA, USA] by setting stringent criterion that included only the experimental findings and excluded data generated from non-neuronal cell lines. Hits were individually subjected to confirmation and upon verification of relevance were included. The data set was used to determine the canonical pathways and the cellular networks.
Imaging, Quantification, and Statistical Analysis
Nikon SMZ1500 and Nikon Eclipse TE2000-E fluorescence microscopes equipped with Intensilight C-HGFI (Nikon, Inc.) were used. Epifluorescence images were acquired using a Digital Sight DS-Qi1MC CCD camera (Nikon, Inc.) and light images were acquired using a Ds-Fi1 camera (Nikon, Inc.). Confocal images were collected using a Zeiss 510 Meta confocal microscope (Carl Zeiss, Inc.).
For behavioral tests, including Rotarod, Gait, and Grip test, statistical differences were determined by one-way ANOVA with post hoc Tukey's multiple comparison parametric tests.
For quantification of CSMN soma diameter, progressive CSMN degeneration, percent PERK expression in Ctip2+ neurons, and percent PDI expression in Ctip2+ neurons, 2 well-defined sections spanning the motor cortex (Section 1: Bregma 0.38 mm, interaural 4.18 mm; Section 2: Bregma −0.34 mm; interaural 3.46 mm; Paxinos and Franklin 2001) were used per mouse.
Numbers and diameters of CSMN were determined in WT and UCHL1−/− mice at P40, P65, P80, and P100. CSMN numbers were counted and averaged (2 sections/per mouse; n = 3). Average CSMN diameter (at least 100 neurons/mouse; n = 3) was measured using the Elements Software (Nikon, Inc., Melville, NY, USA).
To determine the percentage of Ctip2+ neurons that express PERK and PDI, the total numbers of Ctip2+ neurons in layer V of the motor cortex (CSMN) were counted in WT and UCHL1−/− mice at P40, P65, P80, and P100 (n = 3). The Ctip2+ neurons that co-express PERK and PDI were quantified. Statistical differences between WT and UCHL1−/− mice were determined by one-way ANOVA with post hoc Tukey's multiple comparison parametric test.
The presence of total PERK and PDI in CSMN (determined by Ctip2 expression and large soma size) and other cortical neurons were determined by co-immunocytochemistry analysis in vitro using both WT and UCHL1−/− mice. Results were presented as the percent PERK and PDI expression in CSMN and other cortical neurons in the presence of compounds that either enhance (i.e. Tunicamycin and Bortezomib) or reduce (Guanabenz) ER stress. In addition, the percent survival of CSMN and other cortical neurons were quantified using EtHom exclusion. Statistical differences between compound treatments, and survival experiments in WT and UCHL1−/− mice, were determined by the unpaired t-test.
For CSMN spine density measurements, a 50-μm segment of apical dendrites (in layer II/III, in the primary apical dendrite) and basal dendrites (in layer V, separated by 30 μm from the soma) were selected and the total numbers of spines were counted (10 segments/mouse; n = 3). Statistical differences were determined by the unpaired t-test.
All statistical analyses were performed using the Prism software (version 5.0a; Graphpad Software, Inc., La Jolla, CA, USA). Statistically significant differences were determined after either one-way ANOVA with post hoc Tukey's multiple comparison tests or t-test. Statistically significant differences were considered at P < 0.05, and values were expressed as the mean ± SEM.
UCHL1−/− Mice Display Motor Function Defects
To bring an insight on the potential involvement of UCHL1, especially for upper motor neuron biology, we developed a multidisciplinary approach and took advantage of Uchl1nm3419 (UCHL1−/−) mice, which lack all UCHL1 activity (Walters et al. 2008), and display prominent motor function defects (Fig. 1).
UCHL1−/− mice lack part of exon 6 and the intronic sequence between exons 6 and 7 of the Uchl1 gene due to a spontaneous intragenic deletion (Fig. 1A). This mutation results in the creation of a premature stop codon (denoted by "asterisk" in Fig. 1A) resulting in elimination of the transcript. Therefore, functional UCHL1 protein is not detected in UCHL1−/− mice brain (Fig. 1B,C; Walters et al. 2008). UCHL1−/− mice are born at the expected Mendelian ratio, and do not display any phenotypic differences or functional defects at birth and during early development. Heterozygous mice (UCHL1+/−) are viable, fertile with no signs of motor impairment at any age.
DigiGait analysis shows signs of gait impairment by P65: stride (fore limb: WT, 0.34 ± 0.01 s, n = 6; UCHL1−/−, 0.29 ± 0.01 s, n = 7; hind limb: WT, 0.34 ± 0.01 s, n = 6; UCHL1−/−, 0.29 ± 0.01 s, n = 7; P < 0.05, Fig. 1G) and swing (fore limb: WT, 0.14 ± 0.01 s, n = 6; UCHL1−/−, 0.11 ± 0.05 s, n = 7; hind limb: WT, 0.14 ± 0.01 s, n = 6; UCHL1−/−, 0.11 ± 0.01 s, n = 7; P < 0.05). Unlike WT mice, gait parameters of UCHL1−/− mice do not improve with age (Fig. 1G). Although UCHL1−/− pups are similar to their WT littermates at birth and show comparable growth, by P100 all UCHL1−/− mice display profound motor dysfunction (Fig. 1D). There was no difference at P20, but by P40 and onwards hind limb stiffness accompanied with clasping was observed when held upside-down (Fig. 1E; Supplementary Movie 1). UCHL1−/− mice spent progressively less time on the accelerating rotating rod (WT, 4.6 ± 0.1 min, n = 14; UCHL1−/−, 2.8 ± 0.3 min, n = 10; P < 0.001; Fig. 1F). In addition, UCHL1−/− mice gradually lose their ability to hold on to an inverted wire grid (WT, 60 s, n = 4; UCHL1−/−, 27 ± 5 s, n = 6; P < 0.05; Fig. 1H). Even though DigiGait results may suggest the lack of proper gait development, both Rotarod and Grip tests reveal a progressive decline in their motor function with age. These defects are observed with 100% penetrance in the UCHL1−/− mice, and the timing of the progressive motor function defect is comparable among mice and litters. However, both the optic nerve and the cerebellum appear normal, with no signs of apparent degeneration in the UCHL1−/− mice (Supplementary Fig. 1; Bilguvar et al. 2013), suggesting that not all systems are equally affected, and UCHL1−/− mice would be informative for the investigation of UCHL1's role in motor neuron biology.
CSMN Undergo Early, Progressive, and Profound Degeneration in the UCHL1−/− Mice
Retrograde labeling coupled with immunocytochemistry revealed high levels of UCHL1 expression particularly within CSMN in the motor cortex (Supplementary Fig. 2A–D), and initial studies reported high levels of UCHL1 in the cerebral cortex (Day and Thompson 1987, 2010). In addition, UCHL1 was primarily present in Betz cells of postmortem human brain samples (control cases, n = 2; A12–105 and A12–106; Supplementary Fig. 2E). Since CSMN have a unique function for the initiation and modulation of voluntary movement, we next investigated whether CSMN are affected in UCHL1−/− mice. WT and UCHL1−/− motor cortices were comparable with Nissl staining (Fig. 2A,B) and NeuN expression (Fig. 2C,D). Likewise, expression of Lmo4, a molecular marker for CPN (Arlotta et al. 2005), did not show any difference (Fig. 2E,F). However, the expression of Ctip2 (Fig. 2G,H), Crym (Fig. 2I,J), and Fezf2 (Fig. 2K,L), –molecular markers of CSMN in layer V of the motor cortex, and subcerebral projection neuron (SCPN) outside of motor cortex (Arlotta et al. 2005), were reduced at P100, suggesting CSMN degeneration.
To differentiate between cellular degeneration of CSMN and a mere reduction in molecular marker expression, we performed 2 different retrograde labeling experiments (Fig. 3A,B). CSMN were labeled with FG at P30 (Fig. 3A). Their numbers and soma diameter were qualitatively (Fig. 3C,D) and quantitatively (Fig. 3E,F) analyzed at P40, P65, P80, and P100. CSMN morphology, soma diameter, and numbers were similar between WT and UCHL1−/− mice at P40 (Fig. 3C–F). However, by P65, soma diameter (WT, 17.1 ± 0.2 μm; UCHL1−/−, 14.4 ± 0.1 μm, n = 3; P < 0.001, Fig. 3E) and CSMN numbers (WT, 103 ± 3 neurons; UCHL1−/−, 71 ± 2 neurons, n = 3; P < 0.01, Fig. 3F) were significantly different. By P100, there was 88 ± 2% reduction in average CSMN numbers in the UCHL1−/− mice (Fig. 3F). The remaining CSMN had an approximately 50% reduction in soma diameter (Fig. 3E) and displayed the absence of a prominent apical dendrite (Fig. 3D). CPN were retrogradely labeled by contralateral injection of FG at P30, and analyzed at P100 to investigate whether degeneration is restricted to CSMN (Fig. 3B). CPN were comparable between WT and UCHL1−/− mice at P100 (Fig. 3G,H), suggesting that vulnerability is not common to all neurons in the motor cortex.
UCHL1 Function Is Important for the Maintenance of Cytoarchitectural Stability and Protein Homeostasis
To better understand the link between the absence of UCHL1 function and CSMN vulnerability, and to determine the cellular events that would be primarily affected in the absence of UCHL1 function, a system approach was taken. The list of proteins that directly interact with UCHL1 (n = 49) were determined based on a stringent data selection paradigm using IPA. Investigation of the canonical pathways these UCHL1-interacting proteins shared in common, and the molecular networks they activate suggested involvement of UCHL1 primarily with organization of cytoskeleton (n = 21), ubiquitination of proteins (n = 12), and microtubule dynamics (n = 18), apoptosis (n = 29), degradation of proteins (n = 12), and cellular homeostasis (n = 19). Among functions and diseases, movement disorders (n = 18), motor dysfunction (n = 8), and neuromuscular disease (n = 15) were the most prominent. This protein interaction network suggested the potential importance of UCHL1 function in 2 important cellular events: Maintenance of cellular architecture (cytoarchitecture) and protein homeostasis (Supplementary Fig. 3).
Vulnerable CSMN Exhibit Spine Loss with Apical Dendrite Degeneration That Is Not Secondary to CST Defects
To investigate how the absence of UCHL1 function affects cytoarchitecture of CSMN at the soma, apical, and basal dendrites, CSMN were retrogradely transduced with AAV2–2 eGFP (Fig. 4A). AAV transduction did not cause pathological changes, but revealed the details of cytoarchitecture of both WT and UCHL1−/− CSMN (Fig. 4D–O). Transduced WT CSMN exhibited the morphology of healthy CSMN, with large pyramidal cell bodies, prominent apical dendrites, and spines throughout the dendrites, mostly in layers II/III (Fig. 4D,F–I; Jara et al. 2012). In striking contrast, CSMN of UCHL1−/− mice displayed massive disintegration with profound vacuolization and breakdown of cell membrane especially along the apical dendrites (Fig. 4E,J–O; UCHL1−/−, 42 ± 4%, n = 167 apical dendrites, n = 3; P < 0.01; and WT, 7%, n = 281 apical dendrites, n = 3). UCHL1−/− mice had a significant reduction in the numbers of spines throughout the apical (WT, 3.9 ± 0.3; UCHL1−/−, 1.7 ± 0.2; n = 3; P < 0.05, Fig. 4B), but not basal, dendrites (WT, 3.3 ± 0.1; UCHL1−/−, 2.5 ± 0.4; n = 3, Fig. 4C).
Progressive axonal degeneration is suggested as a potential mechanism for CSMN degeneration (Miura et al. 1993; Eisen and Weber 2001; Fischer et al. 2004; Morfini et al. 2013). To investigate if CST defects occur prior to apical dendrite degeneration, and to visualize CST axon fibers, AAV-eGFP was directly injected into the motor cortex (Fig. 5). Large projection neurons were transduced in the motor cortex at P30 and the axons of SCPN of WT and UCHL1−/− mice (n = 3; Fig. 5A) were examined by eGFP expression at the cervical (Fig. 5D), thoracic (Fig. 5E), and lumbar (Fig. 5F) sections of the spinal cord at P60, –when CSMN numbers and soma diameter showed a significant reduction– (Fig. 3E,F). Unilateral injection into the cerebral cortex demonstrated the presence of axons on the left side of the CST in both cervical and lumbar sections of the spinal cord, and further revealed detailed axon morphology, especially on axons that branch and project toward the spinal cord targets (boxed areas enlarged in Fig. 5D′,F′). Axon bundles were observed within the dorsal funiculus with branching toward the anterior and ventral horn (Fig. 5D,F), and they were comparable between WT and UCHL1−/− mice at all levels of the spinal cord. Subtle enlargements along the branches were observed, and in the pons area a subset of axons showed "beads on a string" appearance (UCHL1−/−, 40 ± 6%, n = 114 axons, n = 4; WT, 17 ± 7%, n = 144 axons, n = 4; P < 0.005, Fig. 5B). These results reveal the lack of massive axonal degeneration along the CST at P60, and suggest that apical dendrite defects and spine loss are not secondary to CST degeneration.
ER Stress Is Increased in CSMN of UCHL1−/− Mice
Since UCHL1 is implicated in determining the levels of free ubiquitin, and is important for maintaining protein homeostasis in the cell, we investigated ER stress in CSMN of UCHL1−/− mice. Ubiquitinylated proteins were increased in the motor cortex of UCHL1−/− mice at P100 (Fig. 6A,B). However, PDI, and other markers of ER stress, such as CHOP, BIP, and ATF4, failed to reveal differences between total lysates from motor cortices of WT and UCHL1−/− mice (Supplementary Fig. 4A,B). This could be due to selective increase in a subset of neuron population that cannot be detected when cortical homogenate is used. To investigate if ER stress is specifically increased in CSMN, 2 lines of experiments were performed. First, CSMN were retrogradely labeled with FG. The expression profile of PERK, a major ER stress sensor upregulated upon accumulation of ubiquitinylated and misfolded proteins (Walter and Ron 2011), was studied in retrogradely labeled CSMN (Supplementary Fig. 4C–I). Secondly, Ctip2 expression was used as a marker for CSMN in the motor cortex. PERK (WT, 33% ± 2; UCHL1−/−, 47% ± 4; n = 2) and PDI (WT, 30% ± 7; UCHL1−/−, 45% ± 4; n = 2) immunoreactivity were present especially in FG-labeled CSMN by P65. Even though total numbers of CSMN were decreasing, CSMN (Ctip2+, blue; Fig. 6C,D) with PERK (red; Fig. 6C) significantly increased by P65 (WT, 13.2% ± 1.7; UCHL1−/−, 21.8% ± 2,4; n = 3; P < 0.05; Fig. 6E), and continued to increase with age only in the UCHL1−/− mice. Similarly, PDI expression (red; Fig. 6D) was significantly higher in CSMN of UCHL1−/− mice by P65 (WT, 22.7% ± 0.3; UCHL1−/−, 35.6% ± 1.4; n = 3; P < 0.01; Fig. 6F) and remains significantly higher at older ages (P100 WT, 14.5% ± 1; UCHL1−/−, 39.9% ± 1; n = 3; P < 0.001; Fig. 6F). Albeit total numbers of CSMN reduce with age, the increase in the percentage of CSMN with PERK and PDI suggests a strong correlation between increased ER stress and progressive CSMN degeneration in the UCHL1−/− mice.
CSMN Are Vulnerable to Increased ER Stress
To investigate whether UCHL1−/− CSMN is more vulnerable to increased ER stress, dissociated cortical cultures were established from WT and UCHL1−/− mice at P2–P3. CSMN were identified among other cortical neurons by Ctip2 expression (Fig. 7A–E; red color in the nucleus) and their large soma size (Fig. 7C–E; black arrowheads).
In the first set of experiments, dissociated mixed cultures of WT neurons were exposed to 2 different compounds that increase ER stress via different mechanisms. Tunicamycin blocks N-glycosylation of all proteins and induces ER stress (Olden et al. 1979), whereas Bortezomib induces ER stress by blocking the catalytic domain of the 26S-proteosome (Bonvini et al. 2007; Tuffy et al. 2010). In the second set of experiments, the ER stress was reduced in the dissociated mixed cortical cultures isolated from UCHL1−/− mice by the application of Guanabenz, which enhances the PERK pathway by selectively inhibiting GADD34-mediated dephosphorylation of eIF2α (100 μM; Tsaytler et al. 2011; Vaccaro et al. 2013; Wang et al. 2014). The links between the levels of ER stress, presence of total PERK, PDI, and the percent survival of CSMN in comparison with other cortical neurons were assayed based on selection criteria of PERK, PDI, and Ctip2 expression. Ctip2 expression was restricted to the nucleus (red) and both PERK and PDI were detected in the cell body (blue; Fig. 7A).
In control conditions, both CSMN and other cortical neurons expressed low levels of PERK (CSMN: 25 ± 3%, n = 237 neurons, n = 3; other neurons: 24 ± 5%, n = 134 neurons, n = 3; Fig. 7C,G) and PDI (CSMN, 15 ± 1%, n = 251 neurons, n = 3; other neurons, 17 ± 3%, n = 256, n = 3; Fig. 7D,H). Upon addition of Tunicamycin (100 μM), however, there was an overall increase in the percentage of CSMN that contained high levels of PERK (CSMN, 73 ± 3%; n = 146 neurons; n = 4 mice; other neurons, 48 ± 5%, n = 120 neurons, n = 3; P < 0.005; Fig. 7E,G) and PDI (CSMN, 54 ± 5%, n = 140 neurons, n = 3; other neurons, 31 ± 6%, n = 120 neurons, n = 3; P < 0.05; Fig. 7F,H), and significant differences between CSMN and other cortical neurons were present (Fig. 7G,H).
Addition of Bortezomib (100 nM) increased the presence of PERK and PDI in general, but the percentage of CSMN with PERK (70 ± 2%, n = 167 neurons, n = 4) was significantly higher than other cortical neurons (48 ± 6%, n = 156 neurons, n = 4, P < 0.0001; Fig. 7E,G). Similarly, there was a significant difference between the percentages of CSMN with PDI (47 ± 2%, n = 181 neurons, n = 3) and other neurons (36 ± 2%, n = 141 neurons, n = 3, P < 0.001; Fig. 7F,H). In previous studies, increases in the total levels of PERK and PDI were detected as early as 3 h (Tsaytler et al. 2011). Our current results revealed that, in the presence of increased ER stress, CSMN displayed a selective and profound increase of both PERK and PDI.
Unlike WT CSMN, CSMN of UCHL1−/− mice had intrinsically high levels of total PERK (60 ± 5%, n = 113 neurons, n = 4) and PDI (59 ± 4%, n = 117 neurons, n = 4). This could explain their immediate death upon treatment with Tunicamycin and Bortezomib, and our inability to establish reliable cultures (data not shown). However, other cortical neurons of UCHL1−/− mice had similar levels of PERK (25 ± 1%, n = 236 neurons, n = 3) and PDI (19 ± 5%, n = 245 neurons, n = 3) that are comparable with WT neurons.
Addition of Guanabenz (100 μM) significantly reduced the percentage of CSMN with PERK (40 ± 1%, n = 132 neurons, n = 2, P < 0.05) and PDI (37 ± 1%, n = 143 neurons, n = 2, P < 0.05) within 9 h, but other cortical neurons of UCHL1−/− mice were not affected (PERK; 19 ± 1%, n = 192 neurons, n = 2, P < 0.05 and PDI; 18 ± 3%, n = 180 neurons, n = 2).
These results, together with in vivo expression analyses, suggest that CSMN show selective upregulation of ER stress in the absence of UCHL1 function. This could be one of the underlying causes of CSMN vulnerability. To test the link between increased ER stress and CSMN survival, we performed cell death assay (Fig. 8A,B). Percent survival of CSMN and other cortical neurons of WT mice were similar in control conditions (CSMN, 3 h: 79 ± 2%, n = 283 neurons; 9 h: 75 ± 4%, n = 275 neurons, n = 5; Fig. 8C). In contrast, upon induction of ER stress, more than half of CSMN were dead in culture within 9 h, whereas other cortical neurons were not affected. The percent survival of cortical neurons did not change in the presence of Tunicamycin (3 h: 74 ± 3%, n = 502 neurons; 9 h: 79 ± 1%, n = 396 neurons, n = 5) or Bortezomib (3 h: 77 ± 1%, n = 459 neurons; 9 h: 84 ± 3%, n = 469 neurons, n = 5). However, CSMN of WT mice showed rapid cellular degeneration in the presence of both Tunicamycin (3 h: 64 ± 5%, n = 308 neurons, P < 0.05; 9 h: 40 ± 6%, n = 214 neurons, n = 5, P < 0.001) and Bortezomib (3 h: 67 ± 7%, n = 335 neurons; 9 h: 40 ± 6%, n = 214 neurons, n = 5, P < 0.01). These results suggested that unlike other cortical neurons, CSMN display a unique vulnerability and undergo cellular degeneration in the presence of increased ER stress.
CSMN of UCHL1−/− mice displayed significantly low levels of survival in culture (WT: 85 ± 1%, n = 232 neurons; CSMN: 19 ± 3%, n = 62 neurons, n = 2, P < 0.001; Fig. 8A–C). To test whether reduction in ER stress would improve survival of UCHL1−/− CSMN, cultures were treated with Guanabenz (100 mM) for 9 h (Fig. 8D). In the presence of Guanabenz, CSMN survival significantly improved (38 ± 2%, n = 68 neurons, P < 0.05), whereas survival of other cortical neurons was not affected (76 ± 4%, n = 444 neurons, n = 2; Fig. 8D). These findings show that, in the absence of UCHL1 function, CSMN display selective cellular degeneration, and that increased ER stress is one of the cellular mechanisms responsible for CSMN vulnerability.
UCHL1 is detected in cortical Lewy bodies and neurofibrillary tangles in patients with amyotrophic lateral sclerosis (ALS), PD, Alzheimer's disease (AD), and frontotemporal dementia (Lowe et al. 1990; Alves-Rodrigues et al. 1998; McNaught et al. 2001; Kersaitis et al. 2006; Gong and Leznik 2007). Interestingly, reduced UCHL1 levels are reported in PD, AD, and ALS patients (Choi et al. 2004; Lombardino et al. 2005; Barrachina et al. 2006; Lederer et al. 2007). Two patients with I93M mutation in the UCHL1 gene displayed PD-like symptoms associated with motor dysfunction, stiffness, and tremor (Leroy et al. 1998), and the importance of UCHL1 in the motor system was reinforced with a recent description of UCHL1GLU7ALA mutation in 3 patients displaying an early vision loss followed by progressive neurological dysfunction of the pyramidal system, cerebellum, and spinal dorsal columns, severely affecting movement. Muscle strength was grossly normal, but increased deep tendon reflexes with spasticity indicated prominent upper motor neuron degeneration (Bilguvar et al. 2013).
Mice with varied mutations in the Uchl1 gene (i.e., Uchl1gad mice; Yamazaki et al. 1988; Saigoh et al. 1999) and a targeted deletion-generated murine line, Uchl1tm1Dgen (Chen et al. 2010), have been described. They all report the lack of UCHL1 protein, but interestingly display varied phenotypes. Our goal is not to compare different lines, but to study the role of UCHL1 in motor neuron biology. Uchl1nm3419 (UCHL1−/−) mice, which lack all UCHL1 function (Walters et al. 2008), display major motor function defects without significant sensory ataxia. Unlike the Uchl1gad (Yamazaki et al. 1988; Saigoh et al. 1999) mice, the gracile nucleus is intact in the UCHL1−/− mice. Even though other lines of UCHL1 could be useful for different studies, UCHL1−/− mice offer significant advantages over other currently available lines for the investigation of motor neuron circuitry and disease. Most interestingly, UCHL1−/− mice show similarities to patients with Glu7Ala mutation (Bilguvar et al. 2013), and display early neurodegeneration, with deficits as early as P40. Pathology in the motor cortex of patients is remarkable and CSMN loss is evident by P65 with pronounced degeneration in the UCHL1−/− mice. These observations further support the choice of UCHL1−/− mice to investigate the importance of UCHL1 function for motor neuron biology. Even though CSMN vulnerability has been reported in other mouse models, such as hSOD1G93A mice (Ozdinler et al. 2011) and Prp-TDP-43A315T mice (Wegorzewska et al. 2009), UCHL1−/− mice stand out by their early, progressive, and profound CSMN degeneration that is in part due to ER stress-mediated vulnerability. Interestingly, vacuolation of the apical dendrite and cellular degeneration with spine loss is observed in both UCHL1−/− and hSOD1G93A mice, potentially a common mechanism for cellular pathology (Jara et al. 2013).
CSMN act as a "spokesperson" of the cerebral cortex for the initiation and control of voluntary movement. It is important for CSMN to receive proper input and effective modulation. Since CSMN receive much of their input from layer II/III (Anderson et al. 2010), apical dendrite degeneration, and spine loss, especially in this layer, would impair their ability to receive proper cortical input (Alstermark and Isa 2012). Thus, the message sent to the spinal targets would be cryptic and the motor neuron circuitry would be impaired.
There is a developing need for the generation and characterization of novel tools that mimic human pathology at a cellular level. Failures in clinical trials revealed that the focus has to be on the vulnerable neurons, and not on the extension of lifespan in mice (Genc and Ozdinler 2014). CSMN is of particular importance. Its degeneration is central in various neurodegenerative diseases, especially the ones that involve voluntary movement and motor function defect. Therefore, understanding the reasons behind CSMN vulnerability, and characterization of novel tools with CSMN pathology, would have implications in numerous neurodegenerative disorders, including AD and PD as well as primary lateral sclerosis, hereditary spastic paraplegia, and ALS (Novarino et al. 2014).
Our results suggest a new role for UCHL1 in promoting CSMN health and stability. In the absence of UCHL1 function, CSMN are vastly affected. UCHL1 is previously suggested to mediate cytoarchitectural plasticity by interacting with numerous cellular adhesion complexes, including paxillin, vinculin, and β-catenin (Frisan et al. 2012). Interestingly, Wnt/β-catenin complex contributes to motor neuron pathology in ALS (Yu et al. 2013). UCHL1 has also been reported on the recycling and determining the levels of NCAM (Wobst et al. 2012), one of the best-characterized components of CST. In addition, in the UCHL1-eGFP mouse, a reporter line that expresses eGFP gene under the UCHL1 promoter (Yasvoina et al. 2013), eGFP expression was restricted to CSMN in the motor cortex, further implicating an important role of UCHL1 for CSMN.
We find that CSMN are prone to cellular degeneration in the absence of UCHL1 function, and undergo early and progressive cytoarchitectural defects, especially observed with disintegration of apical dendrites. There is early upregulation of ER stress only in the CSMN of UCHL1−/− mice, and the rate of CSMN survival is proportionally related to increased ER stress. Focusing on the health of vulnerable CSMN would reveal the important information required to develop effective long-term treatment strategies for motor neuron diseases, and diseases in which voluntary movement is impaired.
Authors' Contributions
J.H.J., B.G., E.U., and P.H.O. performed experiments. All authors analyzed data and contributed to the writing of this manuscript.
Supplementary Material
Supplementary material can be found at: http://www.cercor.oxfordjournals.org/.
This work was supported by grants from NIH-R01NS085161-01 (P.H.O.), Les Turner ALS Foundation, Wenske Foundation (P.H.O.), NUCATS (P.H.O. and M.C.B.); NIH/NINDS NS45523, NS49553, and NS41590 (J.D.M.), NIH (5T32AG020506-09 M.A.D. Postdoctoral Training; B.G.), and ALSA Safenowitz fellowship (J.H.J.). Funding to pay the Open Access publication charges for this article was provided by the Les Turner ALS Foundation.
We thank Dr Peter Pytel for the analysis of the human brain samples. D.K. Goncharoff, N.A. Khan, W.D. Weber, M.W. Tu, V. Hernandez, M.J. Standford, M. Schultz, and J. Xie helped with histochemistry and imaging. We thank UPenn Viral Vector Core, B. Goosens for help with confocal microscopy at CMRC Microscopy and Imaging Facility, and NU Behavioral Phenotyping Core Facility. Semi-thin sectioning and toluidine staining were performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. Conflict of Interest: None declared.


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David Cameron and his wife, Samantha, take 100 per cent responsibility for the "error" that resulted in their eight-year-old daughter being left in a country pub.
A spokeswoman for the Prime Minister said the couple were "distraught" when they arrived back at their country residence, Chequers, only to realise that Nancy was not with them.
But she said they did not blame security staff who travelled with the family that day.
"They take responsibility for their own children," she said. "No one is going to face disciplinary action. This was an error. It was a mistake to leave Nancy behind."


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Cultural Competency
The young child population is growing in racial, cultural and linguistic diversity. Children in immigrant families make up a large and growing share of the U.S. child population. CLASP works to ensure that early childhood programs, standards, and policies are responsive to the needs of culturally and linguistically diverse children. We also study the barriers that prevent immigrant families from accessing high-quality child care and early education programs and work to promote policies that remove these barriers and improve access to quality programs.
Feb 3, 2017 | PERMALINK »
Safe Spaces in Early Childhood: New Resources to Support Young Children in Immigrant Families
Last week, the new administration began to issue a series of executive orders on immigration and refugee policy, creating a great deal of uncertainty among immigrant families and the people who serve them. One of the orders focused on interior enforcement would significantly expand immigration enforcement measures, increasing the risk that over 5 million children—the majority of whom are U.S. citizens—could be separated from a parent. Schools and child care and Head Start programs should be prepared to support immigrant families during this difficult time and be aware of the existing protections under current laws and policies.
Last month, the U.S. Department of Education (ED) issued a user-friendly fact sheet to help clarify current policy from the U.S. Department of Homeland Security restricting immigration enforcement actions at or near schools and other "sensitive locations." The fact sheet clearly specifies that child care centers, bus stops, and similar places fall under the "sensitive locations" definition. Another immigration enforcement guide developed by the American Federation of Teachers (AFT) and other partners also proposes how educators and service providers can support and advocate for students and families at risk of being targeted by enforcement actions.
Research consistently shows that fear of separation from a parent due to immigration enforcement hinders a child's ability to concentrate in school and can have harmful long-term implications for a child's overall health and well-being. Heightened concerns over immigration enforcement can also create a chilling effect on school attendance, particularly when local police are engaged in immigration enforcement efforts. Thus, it is important that administrators and providers address the increased anxiety among children and families due to the recent immigration executive orders by assuring them that early childhood programs and schools will remain safe spaces for all students.
Another important tool recently released by ED is a comprehensive resource guide to promote access to quality early learning and elementary education for immigrant families. The guide includes tips for providers on how to better serve children, along with a handbook to educate parents and families about early learning opportunities. The guide also compiles critical information on educational rights of children in immigrant families—such as the federal right to equal access to public education for all children regardless of immigration status—and federal civil rights law that prohibits discrimination on the basis of race, color, national origin, sex, or disability. Finally, it includes an overview of schools' obligations to meet the needs of English Learners and accommodate Limited English Proficient parents.
These resources come at a critical time when our child population is more diverse than ever, yet our policies and systems continue struggling to respond effectively. While young children of immigrants now comprise the fastest growing segment of the U.S. child population, they remain underrepresented in early learning programs, and the recent executive orders now threaten to further deter immigrant families from seeking early learning opportunities.
Schools and program administrators have a responsibility to uphold the educational rights of children in immigrant families, provide families with accurate information, and create welcoming environments where all children can feel safe and supported. CLASP looks forward to working with early childhood advocates to promote programs and policies that will ensure every child has the opportunity to pursue an education free from fear and discrimination. We encourage our partners to share these resources with administrators, providers, and parents.
Read the ED Sensitive Locations Fact Sheet>>
Read the AFT Immigrant and Refugee Children Guide>>
Read the ED Resource Guide: Building a Bright Future for All>>
- Stephanie Schmit and Christina Walker | Feb 17, 2016 Disparate Access: Head Start and CCDBG Data by Race and Ethnicity
- Helly Lee, Christina Walker, Olivia Golden | Dec 14, 2015 Two-Generational Strategies to Improve Family and Child Outcomes
- Emily Firgens and Hannah Matthews | Nov 20, 2012 State Child Care Policies for Limited English Proficient Families
- Stephanie Schmit | Oct 29, 2013 A Strengths-Based Look at the State of the Black Child
- Hannah Matthews and Danielle Ewen | Aug 31, 2010 Early Education Programs and Children of Immigrants: Learning Each Other's Language
- Wendy Cervantes | Feb 03, 2017 Safe Spaces in Early Childhood: New Resources to Support Young Children in Immigrant Families
- Christina Walker and Stephanie Schmit | Dec 13, 2016 A Closer Look at Latino Access to Child Care Subsidies
- Oct 26, 2016 Study Finds Implicit Racial Bias in Preschool Teachers
- CLASP | Aug 16, 2016 2013-2014 Bi-Annual Report
- Jun 02, 2016 Policy Statement Advances HHS and ED's Shared Vision to Improve Access to High-Quality Early Education Opportunities for Dual Language Learners


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- Intraceuticals
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IngredientsTBDWhy We Love IttbdExpert Tips"Obviously, this rejuvenating gel is great for the delicate eye area, but you can also use it to treat fine lines around the lips! "Shipping
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- Members receive Free Standard US Ground Shipping on orders $50 and over. Standard shipping charges apply for non-members.
- Joyus ships to all 50 U.S states. We do not ship to P.O. Boxes, American Samoa, Guam, The Federated States of Micronesia, The Marshall Islands, Northern Mariana Islands, Palau, or The Virgin Islands.
- Return shipping is always on us. Return within the first 30 days, and you'll receive your refund in its original form of payment. Please note that shipping charges are not refundable. After 30 days you'll receive Joyus Dollars to spend on any future purchase.
- Intraceuticals
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Rouge: The Beginning
After years of searching, Robert Haas (Vineyard Brands) and the Perrins (Château de Beaucastel) found the perfect site for their new estate in the steep and stony hills west of Paso Robles. Tablas Creek Vineyard was conceived with vine selections similar to Beaucastel. Natural farming techniques in the vineyard allow our grapes to be the purest expression of the warm climate and the calcareous clay soils in which they grow. Fermenting with native yeasts and selective blending of the varietals, as practiced by Beaucastel for 150 years, allow the wine to best express its rich and elegant esthetic qualities. We have left no stone unturned in our effort to make Tablas Rouge a forceful statement of its outstanding terroir.
Tablas Rouge is estate bottled from vines grown on the Tablas Creek Vineyard Estate cultivated without the use of herbicides or systemic pesticides. Only hand hoeing, cover crops, mulch, scorching and mowing are used for weed control. Natural predatory insects, "Bordeaux Mixture" and sulfur control pests and fungi.
Tablas Creek Vineyard Rouge is modeled after Château de Beaucastel, with a Mourvèdre-based cuvée that combines an appealing earthiness with layers of rich fruit. The harvest was early in 1997, benefitting from a warm spring and a hot, dry summer. Tablas Rouge 1997 shows a very deep cherry red color and presents a ripe berryish nose with leathery overtones. It is complex and layered with jammy red fruit and just a hint of oak. There is good intensity and depth and the wine finishes with excellent persistence on the palate.
Updated tasting notes from a August 2015 tasting can be found on the Tablas Creek blog.
Not Available for Purchase
July 2nd - 9th, 2017
We're excited to be returning to the Mediterranean in the summer of 2017 to host a Rhone River cruise aboard the wonderful Uniworld ship S.S. Catherine. Partner Jason Haas, with his wife Meghan, and Executive Winemaker Neil Collins, with his wife Marci, will host this 8-day cruise from Avignon to Lyon. For all the details, and to book, visit our travel partners' Web site at foodandwinetrails.com/tablascreek2017 or call Food & Wine Trails at (800) 367-5348. We hope that you will join us!
We are excited that we have received our shipping permit for the great state of Arizona. Arizonans can now order wine or join our wine club. More shipping news »
In Antonio Galloni's Vinous (Sept. 2016) 28 Tablas wines topped 90 points, including 2014 Esprit Blanc (93), 2013 Panoplie (95), 2014 Patelin de Tablas (91) and 2014 En Gobelet (93). Read the review » More press »
The CA Sustainable Winegrowing Alliance awarded Tablas Creek one of four 2016 Green Medals for leadership in sustainability. Our practices »
Each month we feature one item that we think is showing particularly well at a 10% discount. February's feature is our 2014 Full Circle Pinot Noir. Details »
February 20, 2017
As many of you know, we have been building up our flock this year. The animals help build up our soil, spreading manure thoroughly and evenly, reducing or eliminating our need to bring in outside fertilizer. They help keep weeds down and reduce the number of tractor passes we need come spring. And they attract different microbes and insects into soil that is vibrantly alive in a way that just doesn't happen in a monoculture. The past few years, we've had around 80 sheep, along with a few alpacas, two donkeys, and a llama. Now, thanks to a fertile winter season, we're up to 165 sheep, plus the other members of the menagerie. The flock can at times be seen from the tasting room, but is more often working quietly, out of view: Read More »


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Head to Old Navy early on Saturday, August 15, 2009 to take advantage of the $2 women's rib knit tank top deal! Limit 5 per customer in select styles.
Thanks Nicole!
Frugal Living Without Sacrifice
By: Briana Cartercomment
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Home > News > Chemistry award announced
September 2nd, 2007
Chemical company Rohm and Haas (NYSE: ROH) has provided an endowment to the American Chemical Society (ACS), supporting the Award for Affordable Green Chemistry. The company, which reported sales of $8.2 billion for 2006, announced in May it would dedicate $150 million — half of its R&D budget — for sustainability efforts, including new product development, energy efficiency and process improvements.
On the heels of the announcement, Rohm and Haas introduced a nanotechnology additive designed to increase the strength of clear cornbased plastics, as well as a formaldehyde-free binder for fiberglass insulation aimed at improving indoor air quality.
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Cutting-edge nanotechnologies are breaking into industries November 18th, 2016
STMicroelectronics' Semiconductor Chips Contribute to Connected Toothbrush from Oral-B That Sees What You Don't: Microcontroller and Accelerometer help brushers clean their teeth more effectively October 4th, 2016
Particle Works launches range of high quality magnetic nanoparticles August 31st, 2016
Sandia use confined nanoparticles to improve hydrogen storage materials performance: Big changes from a small package for hydrogen storage February 25th, 2017
Atom-scale oxidation mechanism of nanoparticles helps develop anti-corrosion materials February 24th, 2017
EmTech Asia breaks new barriers with potential applications of space exploration with NASA and MIT February 22nd, 2017
Molecular phenomenon discovered by advanced NMR facility: Cutting edge technology has shown a molecule self-assembling into different forms when passing between solution state to solid state, and back again - a curious phenomenon in science - says research by the University of Wa February 22nd, 2017
Sandia use confined nanoparticles to improve hydrogen storage materials performance: Big changes from a small package for hydrogen storage February 25th, 2017
New nano approach could cut dose of leading HIV treatment in half February 24th, 2017
Atom-scale oxidation mechanism of nanoparticles helps develop anti-corrosion materials February 24th, 2017
Atomic force imaging used to study nematodes: KFU bionanotechnology lab (head - Dr. Rawil Fakhrullin) has obtained 3-D images of nematodes' cuticles February 23rd, 2017
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Apart from the marble painting mentioned in my previous post we have also been doing a bit more painting this weekend. We tried some stencilling using paper doilies.We painted some Liquid Amber seed pods collected from the front lawn. They look a bit like Christmas decorations so it might be time for Christmas in July...
and we finished painting a treasure box we started at a friend house a couple of weeks ago.
Sunday, July 26, 2009
A bit more painting
Posted by Aussie Mum at 2:08 PM
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Brief historical explanation of the Revelation of st. John, acording to the 'Horć Apocalypticć' of ... E.B. Elliott
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Brief Historical Explanation of the Revelation of St. John, Acording to the ...
Henry Carre Tucker
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Henry Carre Tucker
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Henry Carre Tucker
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abyss Alexander Severus altar Antichrist Apocalyptic apostasy apostle behold bishop Blessed blood Caliph cast century chap Chapter Chris Christendom Christian cloth Commodus Constantine Constantinople council crown Daniel's Danube death denarius Diocletian Divine Domitian dragon earthquake emperor Euphrates expiry faith fell fire four living creatures French Revolution given unto glory God's gospel Goths hand hath heard heathen heaven Heruli holy horns horse imperial judgment kingdoms kings Lamb loud voice mark martyrs Maximian midst mouth nations opened Ostrogoths pagan Papacy Papal persecution plagues Pope preach priests professing prophecy prophetic Reformation reign religion Roman earth Roman empire Rome saints Saracens saying scene Scriptures seal seven angels seven heads shew sitteth slain spirit St John superstition sword symbolised temple thee things third thou throne tion tongue true Church trumpet Turkish vial vision Waldenses whilst wild beast witnesses woman words worship wrath
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Page 111 - And the seventh angel poured out his vial into the air; and there came a great voice out of the temple of heaven, from the throne, saying, It is done.
Page 111 - And he cried mightily with a strong voice, saying, Babylon the great is fallen, is fallen, and is become the habitation of devils, and the hold of every foul spirit, and a cage of every unclean and hateful bird.
Page 99 - And I looked, and behold a white cloud, and upon the cloud- one sat like unto the Son of man, having on his head a golden crown, and in his hand a sharp sickle.
Page 82 - And the woman fled into the wilderness, where she hath a place prepared of God, that they should feed her there a thousand two hundred and threescore days.
Page 7 - And I beheld, and I heard the voice of many angels round about the throne, and the beasts, and the elders: and the number of them was ten thousand times ten thousand, and thousands of thousands; 12 Saying with a loud voice, Worthy is the Lamb that was slain, to receive power, and riches, and wisdom, and strength, and honour, and glory, and blessing.
Page 100 - And the angel thrust in his sickle into the earth, and gathered the vine of the earth, and cast it into the great winepress of the wrath of God.
Page 88 - He that leadeth into captivity shall go into captivity : he that killeth with the sword, must be killed with the sword.
Page 117 - He that is unjust, let him be unjust still: and he which is filthy, let him be filthy still: and he that is righteous, let him be righteous still: and he that is holy, let him be holy still.
Page 100 - King of saints; who shall not fear thee, O Lord, and glorify thy name ? for thou only art holy ; for all nations shall come and worship before thee ; for thy judgments are made manifest.
Page 115 - And he said unto me, It is done. I am Alpha and Omega, the beginning and the end. I will give unto him that is athirst of the fountain of the water of life freely.


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Experience Something Extraordinary in Montgomery County, Maryland
The Victorian Lyric Opera Company (VLOC), a Rockville-based community theater company founded in 1978, will present six performances of Jacques Offenbach's "La Périchole" during two weekends at the F. Scott Fitzgerald Theatre in Rockville. The French comic opera premiered in 1868 in Paris. VLOC's production will be performed in English and accompan View more
After putting together a trilogy of musicals marking the past, present and future of hip-hop culture, local writer and director Psalmayene 24 was looking for a new challenge. He turned to the familiar childhood story of Snow White.
David Benoit has a dream. As a highly-regarded character actor, he can be found on Broadway and off, or crisscrossing the country on regional and national tours. "Jekyll and Hyde," "Phantom of the Opera," "Les Misérables," "Avenue Q"—his resume is lengthy. And when Olney Artistic Director Jason Loewith offered him the opportunity to play the eponym View more
Helen Keller is an American heroine, an icon of strength and accomplishment who overcame tremendous odds to succeed. She was the first deaf and blind woman to earn a bachelor of arts degree and to emerge from almost complete non-verbal isolation to author, activist and lecturer. Triumphs such as these would have been impossible without a voice. It View more
Gaithersburg playwright John Morogiello's work has been performed all over the world, including Theatre 40 in Beverly Hills, Ca. His earliest works—written on his mom's old manual typewriter—were set in less glamorous locales. "She loaded up a piece of paper for me, showed me how it worked, and I immediately started plagiarizing old episodes of 'F View more
Song, dance and a bit of poetry bring to life a fairytale world of magic spells, good and evil, and love and betrayal in the world premiere musical, "Ella Enchanted," on stage at Adventure Theatre MTC in Glen Echo Park.
Ford is one of several child performers in Metro region professional who balance the demands of the theater alongside adolescence and schoolwork. The commitments sometimes require them to leave school early and do additional assignments outside of education hours. "I get everything done sooner or later," Ford said.
Imagine living in a world where everything was the color blue. Azure flowers greet you in the cobalt-colored grass under a turquoise sky. This is the world Inky and Pale have come to know, understand and accept until....they find something red.
It's a classic play, more than 60 years old, but Sandy Spring Theatre Group (SSTG) opens a new production of Reginald Rose's noir courtroom drama "12 Angry Men" at The Arts Barn in Gaithersburg on Jan. 6. "One of the marvelous things is the story itself is timeless," said Bill Spitz, director of the play and SSTG president. "The things that are hap View more
Think you're swamped this holiday season? You should meet Paul Morella. Of course, if you live in Montgomery County, you probably already have.
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3704 S Claridge Rd,Mobile AL, 36608
4 beds , 2 Full/1 Half baths
2,200 sq. ft
Status: Active
Berkshire Hathaway HomeServices Cooper & Co. Inc., REALTORS
(251) 344-5925
Last Updated: 2/21/2017 8:11 AM
Description for 3704 S Claridge Rd Mobile, AL 36608
Beautifully updated home in Yester Oaks/Llanfair. Updated kitchen w/granite countertops, stainless appliances and breakfast area. Seller installed beautiful hardwood floors throughout in 2012 and updated all bathrooms. Formal living room, dining + family room w/built-ins and wood burning fireplace. Plantation shutters throughout. Rannai tankless water heater. Master bedroom has 2 closets, 1 is a walk in. Master bath completely updated w/double sinks, air jet tub and separate, custom shower. Attached double carport with storage and fenced yard. Seller reserves hall bathroom mirror & mounted TV's are negotiable. All updates and improvements are per seller.
Listing Information
Yester Oaks
Electric, Natural Gas
Room Information
Whirlpool Bath
Interior Features
Hardwood, Tile, Carpet Wall to Wall
Central Air, Ceiling Fan(s)
Gas Heat, Central
Exterior / Lot Features
Attached Garage
Wood, Brick
Patio, Rear Porch, Front Porch, Open
Financial Considerations
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Current market conditions for 3704 S Claridge Rd
Active Market Trends
Source:MLS DATA
Median list price$157,900
Days on market136
Properties for sale1,276
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Median sold price$132,000
Sales to list price9,973.0%
Properties sold606
Median sq ft price$76
Arrows indicate 90 day trend
Listing Price History for 3704 S Claridge Rd
Mobile Real Estate
Mobile is located in Alabama. Mobile, Alabama 36608 has a population of 195,239.
The median household income in Mobile, Alabama 36608 is $38,722. The median household income for the surrounding county is $42,973 compared to the national median of $53,046. The median age of people living in Mobile 36608 is 35.6 years.
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FPI Management Inc. - The Marc Palo Alto: redefining urban living. This prestigious downtown community is located near popular University Avenue, restaurants, shopping and vibrant nightlife. Our spacious homes are perfect for every lifestyle and feature gorgeous city views, fully equipped kitchens and air conditioning. Keep life interesting; splash in → Read More
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text: Seller: attictreasures7 (2,567) 100%, Location: Pompano Beach, Florida, Ships to: US & many other countries, Item: 361841094213 Offering For Your Consideration Brand New Sealed 6 Mid Century Wood Pulls. I Think The Wood Is Teak But Not Certain I Only Opened One Of The Packets To Take The Picture. All Of The Pulls Have Screws. The Packaging Is Worn And The Box Is Very Worn. Thanks For Looking And Please Excuse Any Light Glare Just From My Camera Flash. Material: Wood, Color: Medium Wood Tone


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Trump Does Not Deny Campaign in Contact With Russia
He denied personal knowledge or involvement. He also denied ordering Flynn to talk to Russia about sanctions.
Trump Nudges Israel and Palestine Towards Calamity
The White House has yielded to Israel's version of the alt-right.
Rio Olympics Prove Devastating to Brazil
Playing host to the world's largest sporting event isn't such a great deal, after all.
Trump Threatens to "Destroy" Opponents of Civil Forfeiture
For this president, nothing is overreach if the police are doing it.
Democrats Hold a Muted Protest Against Trump's Ban
They're trying to fight the only way they can.


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YP - The Real Yellow PagesSM - helps you find the right local businesses to meet your specific needs. Search results are sorted by a combination of factors to give you a set of choices in response to your search criteria. These factors are similar to those you might use to determine which business to select from a local Yellow Pages directory, including proximity to where you are searching, expertise in the specific services or products you need, and comprehensive business information to help evaluate a business's suitability for you. "Preferred" listings, or those with featured website buttons, indicate YP advertisers who directly provide information about their businesses to help consumers make more informed buying decisions. YP advertisers receive higher placement in the default ordering of search results and may appear in sponsored listings on the top, side, or bottom of the search results page.
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The NACP Review Committee has recently revamped the NACP application process and its forms to incorporate some necessary changes and to address feedback we have received from the field.
We are pleased to announce our new website will launch in January 2017 at www.TheNACP.org Along with the new website, the revised NACP application forms will also be available and will be accepted beginning with the April 1-April 30 application period.
Finally, a change will be made to the NACP application periods. Effective February 1, 2017, applications will no longer be accepted on a rolling basis; the applications will be accepted twice a year during April 1-30 and during October 1-30. Accepting applications on a rolling basis four times per year proved difficult for a volunteer review committee and with a continued effort to keep application fees at a reasonable rate. Those credentialed advocates with expiration dates in February 2018 and August 2018 will have their expiration dates extended to April 30, 2018 and October 31, 2018.
Current application forms may be used for anyone applying for the January 31, 2017 deadline; however, no old forms will be accepted beginning with the April 2017 application period.
We look forward to the new website, your feedback on the revised application forms and to our continuing collaboration to professionalize our advocacy field!
Best wishes for a peaceful and safe holiday season,
The NACP Review Committee
The National Advocate Credentialing Program® (NACP), developed through the cooperative effort of multiple national and state victim assistance organizations, was launched in 2003 as the first voluntary credentialing program available to crime victim advocates nationwide.
NACP is designed to promote a minimum of forty hours of pre-service training for advocates, recognizes the advocate's years of experience, whether volunteer or paid, and requires continuing education for all advocates credentialed under the program.
Four levels of credentialing are offered – Provisional, Basic, Intermediate and Advanced-for all of which eligibility is based upon minimum requirements.
In order to be eligible to apply for Provisional credential, you must have completed a NACP pre-approved forty-hour basic training program. Completion of one of these Pre-approved training programs is also acceptable to meet the "A" Series requirement for all other NACP credential levels (Basic, Intermediate or Advanced). Please click here to see a list of NACP Pre-approved training programs. For more information Contact NACP.
You can submit your application via email, fax, or snail mail:
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text: This topic dominates political discussion on this board. Let's get the truth out. For simplicity I'm grouping two positions together: Against Abortion 1) In all cases 2) Except when the life of the mother is at stake Pro Abortion 1) In all cases 2) In any other case except when the mother's life is at stake. In other words, the Against abortion position only would allow abortion in that fraction of 1% case in which the mother's life is at risk. The Pro Abortion case would allow abortion in other cases.


text:
Joyful Weddings : Anytime
I shot my very first wedding in October 2006. I was a nervous wreck and anxiously paced the length of my living and dining room 2,946 times every day leading to the morning of the wedding. By the time JD and I arrived to the ceremony location, I had gone over every possible situation in my mind and talked JD through a myriad of scenarios as if we were on a CIA co-op. All we needed was the Grassy Knoll.
The day flowed beautifully and I was extremely thankful to have worked with such an awesome bride and groom. One thing I didn't take into consideration was the chance of working with a wonderful wedding coordinator, Courtney Toney of Joyful Weddings and Events. I created a slideshow during the wedding reception and after she saw it, we struck up a conversation, which led to a lovely friendship two years later. Courtney really helped me out in the beginning of my business and sent me some fabulous clients during my first year, for which I was extremely thankful!
I was stoked when Courtney contacted me to photograph her for updated photos of her and her associate, Darcie. We met and wandered the streets of Orange together and had a blast…and just reaffirmed how thankful I am to have met such a wonderful industry peer.
The lovely Courtney Toney…
The lovely Darcie…
Courtney–being the crafty person she is–made these letter cutouts and brought them along to the shoot…
Courtney's one request: A jumping shot…
Happy Sunday!
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November 16th, 2008 at 11:38 pm
I smell me some paper source products….sniff sniff….YUM!
And wowza that's one fantastical jump!
November 16th, 2008 at 11:44 pm
Wow, you've come a long way in just two years. Congratulations! And I love these shots.
November 17th, 2008 at 12:08 am
only 2 years!!! OMG thats amazing 🙂 I did my first real wedding at the same time as you…. I feel so inferior LOL
you are an inspiration to all of us
November 17th, 2008 at 12:20 am
Wow! 2 years. That's amazing. I can't remember what I did before I read your blog first thing in the am. 😉 Totally random question~ and I know you get millions so please forgive me.. but about a year ago you blogged about a cheap video light that you use.. {it lasts for like 30 minutes and is rechargeable. } I've been searching your blog all day and can't seem to find the wedding that you mentioned it. If it's not too much trouble.. What video light is it??
November 17th, 2008 at 12:32 am
So glad I found your blog. Love the suitcase shots! LOVE EM!
You ROCK!
November 17th, 2008 at 12:42 am
Not sure why, but I always love people jumping in the air. It just seems so happy!
November 17th, 2008 at 12:47 am
Wow, it's hard to believe you've only been shooting weddings for 2 years! I love the one on the vintage suitcases!!
November 17th, 2008 at 12:48 am
Oh wow! SO CUTE. I totally sympathize with your pre-wedding jitters. I had NIGHTMARES about mine. I'll have to blog about it soon. Haha, that was back in May, only a few months ago. Sheesh. Scariness. It's a little easier now. 😉
November 17th, 2008 at 12:48 am
I just had my first wedding this past weekend and I too was a nervous wreck. But it went well. I think I could get used to it.
November 17th, 2008 at 1:16 am
I love these gals! Their work is fantastic. Great shots, Jasmine!
November 17th, 2008 at 1:32 am
We love you girl! Thanks so much- these are so great! I am so thankful that our paths crossed- I cannot imagine these past couple of years in the industry without you.
Funny note: David laughed really hard at the last one and told me it was scandalous… oops! I guess that one won't be going on the website 🙂 I am highly impressed with the height I got will all 5 ft of myself, though!
November 17th, 2008 at 3:41 am
Very very cool set! I love the last one
November 17th, 2008 at 3:53 am
I love your blog! It makes me happy coming here everyday. And I especially love that you are helping me find all my wedding vendors ahead of time…now all I need is a man! =-)
November 17th, 2008 at 4:02 am
these are so adorable jasmine! courtney just makes me happy, she's the biggest sweetheart ever. love the suitcases!!!
November 17th, 2008 at 4:29 am
Oh my, I have not done a wedding yet and I can so related with the anxiety you had felt, cause weddings is a hit or miss, you have seconds to capture very important parts of the ceremony and key moments, any tips for a first timer would be grealty humbly appreciated, love the fun pictures of the girls.
November 17th, 2008 at 5:03 am
I did my first wedding in March of this year and I felt exactly the same way!
November 17th, 2008 at 6:33 am
Courtney looks GORGEOUS! Jasmine, what a way to capture her spirit!! Even as a fellow coordinator, Courtney's "joyful"ness comes out constantly. I can only imagine what she's like to work with!! I love these shots of her and Darcie!
November 17th, 2008 at 6:40 am
YOU got a workout on that first wedding. ha!
November 17th, 2008 at 7:03 am
I know I shouldn't be surprised by now, but I'm always amazed at how you capture light! Not just the stuff from the sun, the the inner light in people. I think you just have a way of bringing it out in people, and your camera just happens to be around to capture it!
November 17th, 2008 at 9:42 am
It also reminds me of my first wedding. I couldn't sleep at night and had to go to the bathroom 3 times before leaving my place
November 17th, 2008 at 12:47 pm
you should write a book! I would love to hear what all you did leading up to that first wedding to become such a flippin' awesome photographer!
November 17th, 2008 at 1:17 pm
Love the suitcases by the brick wall shot 🙂
November 17th, 2008 at 2:42 pm
LOVE the "joyful" letters. she is super crafty! :o) great pics. I too wish one day for a "jumping" shot. hehe
November 17th, 2008 at 2:46 pm
Rockin photos!! Love the jumping one, she got some serious air! and with heels!!! Way to go!
November 17th, 2008 at 2:50 pm
LOVE the suitcases in this shoot! What a cute and quirky little prop 😉
November 17th, 2008 at 3:18 pm
I love this series! especially the last one with her jumping! so much fun!
November 17th, 2008 at 3:45 pm
great shots jasmine. anytime….. 🙂
November 17th, 2008 at 3:57 pm
Ooooo I like the suitcase turned to sitting area idea! I LOVE HOW YOU THINK! You are so creative!
November 17th, 2008 at 5:11 pm
They look so fab J*. I really like the ones of them together. They look great! These are head shots anyone would be proud to use.
November 17th, 2008 at 5:31 pm
My favorite is the jumping shot!
November 17th, 2008 at 6:56 pm
Because of your blog, I'm starting to believe that everyone in CA is just beautiful! 🙂 Great job as always!
November 17th, 2008 at 7:33 pm
I been wanting to so a shoot in downtown orange.
November 17th, 2008 at 9:26 pm
adore courtney, adore you, and adore these pics!! xoxo
November 17th, 2008 at 9:32 pm
Awesome! Do you bring your own props like the suitcases or do your clients bring them along?
November 17th, 2008 at 10:13 pm
courtney is the best wedding coordinator! jasmine-you did an amazing job with these photos! she looks gorgeous! everyone should book you two for their wedding!
November 18th, 2008 at 12:22 am
Love the suitcases!! Great shots!
November 18th, 2008 at 8:32 am
Here's another vote for the suitcases [since I collect vintage suitcases], I thought that was brilliant! But they are all terrific!
November 18th, 2008 at 3:35 pm
Love the use of the suitcases and the last photo of Courtney jumping. She got some height on that one.
November 18th, 2008 at 4:30 pm
Great jump shot!!! …well great set all around really!
November 24th, 2008 at 3:25 am
those "joyful" letters are such a "courtney touch"! i love it!
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