Depletion of COPI in cancer cells: the role of reactive oxygen species in the induction of lipid accumulation, noncanonical lipophagy and apoptosis.

The coatomer protein complex 1 (COPI) is a multisubunit complex that coats intracellular vesicles and is involved in intracellular protein trafficking. Recently we and others found that depletion of COPI complex subunits zeta (COPZ1) and delta (ARCN1) preferentially kills tumor cells relative to normal cells. Here we delineate the specific cellular effects and sequence of events of COPI complex depletion in tumor cells. We find that this depletion leads to the inhibition of mitochondrial oxidative phosphorylation and the elevation of reactive oxygen species (ROS) production, followed by accumulation of lipid droplets (LDs) and autophagy-associated proteins LC3-II and SQSTM1/p62 and, finally, apoptosis of the tumor cells. Inactivation of ROS in COPI-depleted cells with the mitochondrial-specific quencher, mitoquinone mesylate, attenuated apoptosis and markedly decreased both the size and the number of LDs. COPI depletion caused ROS-dependent accumulation of LC3-II and SQSTM1 which colocalizes with LDs. Lack of double-membrane autophagosomes and insensitivity to Atg5 deletion suggested an accumulation of a microlipophagy complex on the surface of LDs induced by depletion of the COPI complex. Our findings suggest a sequence of cellular events triggered by COPI depletion, starting with inhibition of oxidative phosphorylation, followed by ROS activation and accumulation of LDs and apoptosis.

Numerical method for the projected Gross-Pitaevskii equation in an infinite rotating two-dimensional Bose gas.

We present a method for evolving the projected Gross-Pitaevskii equation in an infinite rotating Bose-Einstein condensate, the ground state of which is a vortex lattice. We use quasiperiodic boundary conditions to investigate the behavior of the bulk superfluid in this system, in the absence of boundaries and edge effects. We also give the Landau gauge expression for the phase of a BEC subjected to these boundary conditions. Our spectral representation uses the eigenfunctions of the one-body Hamiltonian as basis functions. Since there is no known exact quadrature rule for these basis functions we approximately implement the projection associated with the energy cutoff, but we show that by choosing a suitably fine spatial grid the resulting error can be made negligible. We show how the convergence of this model is affected by simulation parameters such as the size of the spatial grid and the number of Landau levels. Adding dissipation, we use our method to find the lattice ground state for N vortices. We can then perturb the ground-state, to investigate the melting of the lattice.

Striated myogenesis and peristalsis in the fetal murine esophagus occur without cell migration or interstitial cells of Cajal.

Esophageal striated myogenesis progresses differently from appendicular myogenesis, but the mechanism underlying this process is incompletely understood. Early theories of transdifferentiation of smooth muscle into striated muscle are not supported by transgenic fate-mapping experiments; however, the origin of esophageal striated muscle remains unknown. To better define the process of striated myogenesis, we examined myogenesis in murine fetal cultured esophageal whole-organ explants. Embryonic day 14.5 (E14.5) esophagi maintained a functional contractile phenotype for up to 7 days in culture. Striated myogenesis, as evidenced by myogenin expression, proceeded in a craniocaudal direction along the length of the esophagus. Esophageal length did not change during this process. Complete, but not partial, mechanical disruption of the rostral esophagus inhibited myogenesis distally. Addition of fibroblast growth factor-2 (FGF-2) to the culture media failed to inhibit striated myogenesis, but attenuated smooth muscle actin expression and reduced peristaltic activity. Inhibition of c-kit failed to inhibit peristalsis. These results suggest that striated myogenic precursors are resident along the entire length of the esophagus by day 14.5 and do not migrate along the esophagus after E14.5. Induction of myogenesis craniocaudally appears to require physical continuity of the esophagus and is not inhibited by FGF-2. Finally, peristalsis in E14.5 esophagi appears not to be regulated by interstitial cells of Cajal.

Posterior capsulotomy and retinal detachment following extracapsular lens surgery.

A retrospective analysis of the results of extracapsular cataract surgery with and without implant is presented. Surgery was performed by a group of consultants and registrars from 1975 to 1982, and minimum patient follow-up was 12 months (mean 26.2 months). The incidence of postoperative capsulotomy was 14.3%, but the need for this procedure declines, becoming negligible after 3.5 years. Of 242 eligible eyes, aphakic retinal detachment occurred in only one.

A device for automated laser delivery in retinal photocoagulation.

We report a modification to the argon-krypton laser (Lasertek Model AK 41) which allows optional automatic operation. This is a safe, time-saving device for use in cases requiring extensive retinal laser treatment.

Extracapsular cataract extraction - some problems.

Comments are made on the method of removing the anterior capsule during the performance of extracapsular cataract operations. The difficulty of performing this manoeuvre adequately is emphasised. Observations carried out during the postoperative period by means of the slit-lamp revealed some hitherto undescribed changes in the capsule. The edge of the anterior capsule can be clearly seen and looks like broken glass. Fine fibrils have been observed attached to the fractured capsular edge. In the early postoperative period the anterior capsule tends to be separated from the posterior by quite a deep space, but at an early stage, usually about the fourth day, the edge of the anterior capsule becomes adherent to the posterior capsule, and rather rapidly a dense fibrous line forms along the joint.

The usefulness of Vistech and FACT contrast sensitivity charts for cataract and refractive surgery outcomes research.

AIM: To investigate the repeatability and sensitivity of two commonly used sine wave patch charts for contrast sensitivity (CS) measurement in cataract and refractive surgery outcomes. METHODS: The Vistech CS chart and its descendant, the Functional Acuity Contrast Test (FACT), were administered in three experiments: (1) Post-LASIK and age matched normal subjects; (2) Preoperative cataract surgery and age matched normal subjects; (3) Test-retest repeatability data in normal subjects. RESULTS: Contrast sensitivity was similar between post-LASIK and control groups and between the Vistech and FACT charts. The percentage of subjects one month post-LASIK achieving the maximum score across spatial frequencies (1.5, 3, 6, 12, 18 cycles per degree) were (50, 33, 13, 13, 0 respectively) for FACT, but only (0, 0, 13, 4, 0 respectively) for Vistech. A small number of cataract patients also registered the maximum score on the FACT, but up to 60% did not achieve the minimum score. Test-retest intraclass correlation coefficients varied from 0.28 to 0.64 for Vistech and 0.18 to 0.45 for FACT. Bland-Altman limits of agreement across spatial frequencies were between +/-0.30 and +/-0.85 logCS for Vistech, and +/-0.30 to +/-0.75 logCS for FACT. DISCUSSION: The Vistech was confirmed as providing poorly repeatable data. The FACT chart, likely because of a smaller step size, showed slightly better retest agreement. However, the reduced range of scores on the chart due to the smaller step size led to ceiling (post-LASIK) and floor (cataract) effects. These problems could mask subtle differences between groups of patients with near normal visual function as found post-refractive or cataract surgery. The Vistech and FACT CS charts are ill suited for refractive or cataract surgery outcomes research.

Indirect choroidal tears at the posterior pole: a fluorescein angiographic and perimetric study.

Retinal fluorescein angiographic and visual field studies were performed on 10 patients who had developed indireect choroidal tears, these procedures being repeated at intervals until the retinal disturbances stabilised. Fluorographic investigations revealed that a transient breakdown of the choroidoretinal barrier to fluorescein dye could be detected in cases investigated within 4 days of injury, and also that the late complication of neovascularisation, producing a serous maculopathy, may resolve and good central vision be retained without recourse to photocoagulation therapy. Patients who had visual field examinations performed within a few days of trauma were noted to have dense central scotomata, but some recovery occurred in most cases, although to a variable degree. Field defects away from fixation corresponded to an extent with areas of post-traumatic pigmentary retinopathy, but in some portions of retina with normal background appearances sensitivity was also noted to be reduced. Scotomata corresponding specifically to choroidal tears or nerve fibre bundle defects were not recorded. Improvement of visual acuity may be expected, except in cases where a choroidal tear has involved the fovea or progressive choroidal neovascularisation led to permanent macular damage.

Microsporidial keratoconjunctivitis in a patient with AIDS.

A male patient is described with acquired immune deficiency syndrome (AIDS) who developed chronic keratoconjunctivitis and chronic sinusitis due to infection with the microsporidian Encephalitozoon cuniculi. Diagnosis was confirmed by electron microscopic examination of conjunctival epithelial cells and nasal polypectomy specimens. Treatment with propamidine isethionate 0.1% (Brolene) eye drops six times daily led to a prompt resolution of the keratoconjunctivitis.

Learning effects of a back education program.

STUDY DESIGN: This study involved a post-test only, control group design. OBJECTIVES: To analyze the learning effects of back education programs (video and classroom learning). SUMMARY OF BACKGROUND DATA: Previous research has examined lost work time and workers compensation costs but has not addressed the learning effects of back schools. This study used the American Back School as the education intervention. The American Back School teaches students to maintain the lumbarlordosis while lifting. METHODS: The subjects (n = 205) were assigned to three groups through modified randomization. Three employees who previously sustained low back injury were placed in the back school group. The back school group, Group 1, (n = 74) attended a back school program that included cognitive learning strategies and practice in correct lifting. A video group, Group II, (n =64) viewed a similar program that consisted of spinal anatomy and biomechanics and instruction in correct lifting technique. A control group, Group III, (n = 67) received no back education. One week after the education intervention, 145 of the subjects from the three groups had the lumbar lordosis measured with a flexible ruler while assuming a lifting position. The ruler was placed over the lumbar spinous processes, and the lordotic angle was calculated A 12-item multiple choice test and a 10-item. Likert scale were administrated to 199 of the subjects in the three groups to determine the cognitive learning effect and the perceived relevance of the program, respectively. RESULTS: Multivariate analysis of variance was used and demonstrated significant differences between the back school group and the control group on the cognitive, psychomotor, and affective measures at the 0.001 level. No significant differences were found between the video and control groups on the measures with additional univariate testing. CONCLUSIONS: The results indicate that the back school is an effective tool for influencing lifting posture and conveying information regarding spinal mechanics and lifting technique. In addition, the back school videos may not be an effective means of preventing low back injury.

Survival, intracranial lesions, and neurodevelopmental outcome in infants with congenital diaphragmatic hernia treated with extracorporeal membrane oxygenation.

OBJECTIVE: Before the use of extracorporeal membrane oxygenation (ECMO), infants with a severe form of congenital diaphragmatic hernia (CDH) had a high mortality and morbidity. Recent studies have shown an improvement in the survival of these infants after ECMO treatment; however, the existing data do not provide sufficient informations regarding the quality of survival and developmental outcome of these infants. The objective of this study was to evaluate survival, intracranial lesions, and the neurodevelopmental outcome of infants with CDH treated with ECMO. METHODS: We retrieved data for 51 (n = 51) infants with CDH who were treated with ECMO at Huntington Memorial Hospital between 1985 and 1994. Their mean gestational age was 38.5 +/- 2.4 weeks (mean +/- SD); their mean birth weight was 3170 +/- 620 gm. Vital signs, arterial blood gases, chest radiographs, cranial and cardiac ultrasonography were routinely obtained before ECMO treatment. Cranial ultrasounds were performed daily on all infants while on ECMO; computerized tomography scans were obtained on all infants after completion of ECMO treatment. The surviving infants were followed at our neonatal follow-up clinic for neurodevelopmental assessment. RESULTS: A total of 39 infants were placed on venoarterial ECMO and 12 infants were placed on venovenous ECMO; a total of 35 infants had CDH repair before ECMO, whereas 16 infants had delayed surgery. A total of 31 infants (61%) survived. The infants who survived had a mean pH of 7.33 +/- 0.20, mean airway pressure of 19.6 +/- 5.8 cm H2O, and an oxygenation index (OI) of 87 +/- 55 before ECMO intervention. The infants who expired (n = 20) had a mean pH of 7.31 +/- 0.15, mean airway pressure of 23.1 +/- 5.5 cm H2O, and a mean oxygenation index of 127 +/- 56 before ECMO treatment. Before ECMO, survivors had a significantly lower oxygenation index and a higher Pao2 compared with nonsurvivors (p < 0.01). A total of 18 infants (35%) had abnormal central nervous system findings. Of the 51 infants, 10 had ventricular dilatation, 6 had intracranial hemorrhage, and 11 had focal or diffuse cerebral atrophy diagnosed by computerized tomography scan or at autopsy (1 patient had an infarct). Eight infants had more than one central nervous system abnormality. A total of 16 survivors had a neurodevelopmental evaluation at 12 months, and 11 of these survivors were evaluated at 24 months of age (Bayley Scales of Infant Development). The developmental progress of these infants falls within the low-average range of cognitive and motor abilities. Their mean Bayley Mental Developmental Index was 85 +/- 25 (50 to 145) at 24 months; their Psychomotor Developmental Index was 89 +/- 21 (50 to 113) at 24 months of age. Follow-up at 4 and 6 years of age is in progress. CONCLUSION: Our preliminary findings indicate that 35% of infants with severe CDH requiring ECMO had central nervous system abnormalities (intracranial lesions, including ventricular dilatation). The survival rate in our study population is consistent with recent reports. As a group, infants with severe CDH display mild neuromotor and cognitive delay in development at 24 months of age.

Design, analysis, and implementation of a parallel tree search algorithm.

The alpha-beta algorithm for searching decision trees is adapted to allow parallel activity in different parts of a tree during the search. The algorithm has been implemented in a procedural simulation language (GASP IV). The simulation environment provides the illusion of multiple software processes and multiple hardware processors. A number of preliminary experiments have been done to gather statistics on run time, nodes scored, and nodes visited. Results indicate that a substantial reduction in time of search occurs because of the use of parallelism. An analytic expression for the storage requirements of the algorithm is derived. The analysis provides an example of the classical tradeoff between time and storage.

Use of stapling instruments to aid in the removal of ovarian tumours in mares.

Ovarian granulosa theca cell tumours were removed surgically in 10 mares. Stapling instruments were used to facilitate haemostasis of the ovarian pedicle in five out of 10 cases and as a sole method of haemostasis in the remaining five cases. Postoperative haemorrhage from the ovarian pedicle was suspected in only one case. All cases survived long term follow-up from six months to four years. Results from this series of cases indicate that stapling instruments provide a rapid, easy and efficient means of ligating ovarian pedicles when performing ovariectomy in the mare.

Comparative effects of cholera toxin, Salmonella typhimurium culture lysate, and viable Salmonella typhimurium in isolated colon segments in ponies.

Isolated segments of left dorsal colon and a side-to-side colocolostomy (between the left ventral colon and left dorsal colon) were surgically created in 6 adult ponies. Four segments, each separated by an empty segment, were inoculated (20 ml) with 1 of the following 4 solutions: phosphate buffered saline solution (PBSS)/1% polyethylene glycol (PEG); purified cholera toxin in PBSS/1% PEG (5 micrograms cholera toxin/ml of PBSS/1% PEG); lyophilized Salmonella typhimurium UCD 1755 culture lysate, reconstituted in PBSS/1% PEG; and viable S typhimurium UCD 1755 (10(8) organisms/ml of PBSS/1% PEG). Twenty hours following inoculation of the treatment solutions into the isolated colon segments, the ponies were reanesthetized. Fluid accumulation in the isolated segments was measured, and tissue samples from isolated segments were taken for examination by light microscopy and electron microscopy, and for measurement of mucosal cyclic adenosine monophosphate levels. There was fluid accumulation in segments inoculated with cholera toxin in 4 ponies (29.5 +/- 12.7 ml), and in segments inoculated with S typhimurium UCD 1755 culture lysate in 3 ponies (14.0 +/- 8.7 ml). There was no fluid accumulation in segments inoculated with either the control solution (PBSS/1% PEG) or viable S typhimurium UCD 1755. There was significantly (P less than 0.05) less cyclic adenosine monophosphate in segments inoculated with cholera toxin, Salmonella lysate, and viable Salmonella, compared with control segments. Histologically, there were minimal changes in control segments, consisting of mild to moderate submucosal edema and capillary congestion.(ABSTRACT TRUNCATED AT 250 WORDS)

Duodenal leiomyoma associated with colic in a two-year-old horse.

A 2-year-old horse with signs of colic, reflux on nasogastric intubation, and large colon distention on transrectal palpation was found to have a leiomyoma involving the duodenum. The mass was excised without penetration of the lumen of the small intestine, and the horse recovered without complications.

Use of a bone plate for treatment of middle phalangeal fractures in horses: seven cases (1979-1984).

Four adult horses and 3 foals with middle phalangeal fractures were treated by arthrodesis of the proximal interphalangeal joint, using a bone plate. Six of the 7 horses survived greater than 2 years; 2 of the 6 horses had intermittent lameness after hard work, and 4 horses didn't have evidence of lameness. The use of a bone plate for arthrodesis of the proximal interphalangeal joint was a successful treatment alternative for middle phalangeal fractures in horses.

Secondary exotropia: a retrospective analysis of matched cases.

Secondary exotropia, that is exotropia following surgery for esotropia, remains frustratingly common, even among experienced strabismologists. In this study, we have compared a group of secondary exotropia cases with a matched group of cases of residual esotropia. Of the factors analyzed, only the presence of an A- or V-pattern preoperatively (P = .0077), the presence of limited postoperative adduction (P < .001), and postoperative binocular function (P = .0073) identified the secondary exotropes. The presence of reduced adduction postoperatively in subjects with secondary exotropia was a poor prognostic sign with 11 of the 13 cases requiring further surgery for poor cosmesis. The authors suggest two subclassifications of secondary exotropia.

Adjustable and non-adjustable strabismus surgery: a retrospective case-matched study.

Adjustable suture strabismus surgery was introduced to improve results in unpredictable strabismus cases. However, its usage is increasing and in some centres is routine. The present authors propose that the routine use of the adjustable suture technique can only be justified if it can be shown to confer an advantage to the patient. In a retrospective analysis of matched data, they compared the results of their adjustable with non-adjustable strabismus surgery. The adjustable suture procedures performed during a 5-year period, on non-thyroid eye disease patients, were matched to non-adjustable cases according to the type and aetiology of strabismus and the magnitude of deviation. Pairs were matched as closely as possible according to age and strabismus surgery history. All cases were incomitant. The surgical results of the two groups were analysed with regard to the pre-operative and post-operative angles of deviation, the post-operative drift, and a successful outcome, pre-defined by carefully selected criteria. Twenty-six cases were analysed in each of the two groups. Mean pre- and post-operative angles of deviation showed no significant difference between the two groups. 'Success' rates were 81% in the adjustable group and 88% in the non-adjustable group. Given that the success rate of the two techniques is similarly high, a much larger study is required to detect a difference in results, with statistical significance. The authors conclude that there is currently insufficient evidence that patients, without thyroid eye disease, benefit from the longer and potentially uncomfortable procedure of adjustable suture strabismus surgery to support its rapidly growing use and that a prospective randomised controlled trial is indicated.