A panel of eGFP reporters for single base editing by APOBEC-Cas9 editosome complexes.

The prospect of introducing a single C-to-T change at a specific genomic location has become feasible with APOBEC-Cas9 editing technologies. We present a panel of eGFP reporters for quantification and optimization of single base editing by APOBEC-Cas9 editosomes. Reporter utility is demonstrated by comparing activities of seven human APOBEC3 enzymes and rat APOBEC1 (BE3). APOBEC3A and RNA binding-defective variants of APOBEC3B and APOBEC3H display the highest single base editing efficiencies. APOBEC3B catalytic domain complexes also elicit the lowest frequencies of adjacent off-target events. However, unbiased deep-sequencing of edited reporters shows that all editosomes have some degree of local off-target editing. Thus, further optimization is required to generate true single base editors and the eGFP reporters described here have the potential to facilitate this process.

Revisiting early hypothyroidism screening in infants with Down syndrome.

OBJECTIVE: To identify if the incidence of hypothyroidism in infants with Down syndrome is higher than previous childhood estimates (15%) when examined prior to the standard retesting at 6 months of age. STUDY DESIGN: A retrospective observational cohort study of 122 children with Down syndrome admitted to a university-based birthing hospital between May 2000 and March 2012. Demographic data (for example, date of birth, gender, gestational age, inborn) and diagnostic data (Down syndrome, congenital heart disease and gastrointestinal disease) were cross-linked with thyroid hormone laboratory tests (total thyroxine, free thyroxine and thyroid stimulating hormone) to determine incidence of identified hypothyroidism and thyroid testing prior to 4 months of age (n=80). RESULT: In all, 32.5% were found to have any hypothyroidism. Of these, 14 were primary hypothyroidism (17.5%) needing supplemental T4 therapy, 12 were compensated hypothyroidism (15%) and euthyroid was identified in 54 infants (67.5%). CONCLUSION: Despite normal newborn screens, the incidence of any hypothyroidism (early compensated hypothyroidism and primary hypothyroidism) was higher than previously reported.

Local plasticity of Al thin films as revealed by x-ray microdiffraction.

Grain-to-grain interactions dominate the plasticity of Al thin films and establish effective length scales smaller than the grain size. We have measured large strain distributions and their changes under plastic strain in 1.5-microm-thick Al 0.5% Cu films using a 0.8-microm-diameter white x-ray probe at the Advanced Light Source. Strain distributions arise not only from the distribution of grain sizes and orientation, but also from the differences in grain shape and from stress environment. Multiple active glide plane domains have been found within single grains. Large grains behave like multiple smaller grains even before a dislocation substructure can evolve.

Scanning X-ray microdiffraction with submicrometer white beam for strain/stress and orientation mapping in thin films.

Scanning X-ray microdiffraction (microSXRD) combines the use of high-brilliance synchrotron sources with the latest achromatic X-ray focusing optics and fast large-area two-dimensional-detector technology. Using white beams or a combination of white and monochromatic beams, this technique allows for the orientation and strain/stress mapping of polycrystalline thin films with submicrometer spatial resolution. The technique is described in detail as applied to the study of thin aluminium and copper blanket films and lines following electromigration testing and/or thermal cycling experiments. It is shown that there are significant orientation and strain/stress variations between grains and inside individual grains. A polycrystalline film when investigated at the granular (micrometer) level shows a highly mechanically inhomogeneous medium that allows insight into its mesoscopic properties. If the microSXRD data are averaged over a macroscopic range, results show good agreement with direct macroscopic texture and stress measurements.

Efficacy and durability of ultraviolet tints in CR-39 ophthalmic lenses.

BACKGROUND: Ocular protection from solar ultraviolet (UV) radiation has been emphasized in recent years as a result of the thinning of the ozone layer in the atmosphere. The purpose of this study was to evaluate the absorptive properties of UV tints in CR-39 lenses. METHODS: We used a spectrophotometer to measure the UV transmittance of three groups of UV tinted CR-39 lenses, including (1) lenses tinted by local optical laboratories: (2) lenses tinted by us, using commercially available dyes: and (3) stock UV lenses that have UV absorptive molecules throughout the lens. We also tested the durability of these tints to daily washing/drying by measuring their UV transmittance characteristics at 3, 6, and 12 months. RESULTS: All the tested lenses absorbed all of the UV-B and at least 99% of UV-A. The durability of these UV tints when exposed to daily washing/drying was excellent: all lenses continued to absorb all of the UV-B and at least 99% of UV-A after 1 year. CONCLUSIONS: These data suggest that UV tinted CR-39 lenses provide protection against UV radiation that meets the ANSI Z80.3-1996 Standard for non-prescription sunglasses and fashion eyewear. Furthermore, normal daily washing/drying for 1 year does not cause a significant decrease in the protective effect of the UV tint.

Cell-specific delivery of bacteriophage-encapsidated ricin A chain.

We have used covalent coupling of deglycosylated ricin A chain (RAC) to the assembly initiation/translational repression RNA stem-loop (TR) of the bacteriophage MS2 to direct encapsulation of the toxin in bacteriophage capsids. Multiple copies of the TR-RAC conjugate can be incorporated into single capsid shells. The resultant particles can then be directed to specific cells by receptor-mediated endocytosis (RME) of complexes formed with anti-MS2 coat protein antibodies or by further covalent modification of the capsids by addition of human transferrin molecules. The results suggest that bacteriophage encapsulation and targeting is an efficient way to deliver toxins in a cell-specific fashion. The system may have widespread application in the field of targeted drug delivery, including antisense reagents.

Experience with early postcesarean hospital dismissal.

OBJECTIVE: Our purpose was to assess early postcesarean hospital dismissal. STUDY DESIGN: A retrospective review was performed of all women receiving cesarean delivery over the most recent 6-month period in a busy private obstetrics practice that routinely dismisses its cesarean patients on postoperative day 2. Women who meet certain criteria (uncomplicated pregnancy, Pfannenstiel incision, uncomplicated surgery, no febrile morbidity, stable vital signs, ability to ambulate without assistance, ability to urinate without assistance, and auscultation of active bowel sounds) on postoperative day 2 are dismissed from the hospital. Outcomes were compared against women undergoing cesarean delivery during the 6 months immediately before the institution of the early dismissal program. RESULTS: Among 147 women undergoing cesarean deliveries, 117 (80%) met the criteria for early dismissal. When compared with controls (n = 93), there was no difference in outcomes. No one in the early dismissal group required readmission to the hospital. CONCLUSION: Among properly selected candidates, early postcesarean hospital admission is a reasonable option.

Underrecognition of tardive dyskinesia and drug-induced parkinsonism by psychiatric residents.

Recognition of tardive dyskinesia (TD) and other neuroleptic, drug-induced, extrapyramidal side effects presents a major challenge in modern clinical psychopharmacology. Failure to recognize these disorders can lead to poor patient care and may contribute to societal pressure for external control of psychiatric practice. This study reports the occurrence of tardive dyskinesia and drug-induced parkinsonism (DIP) in 101 inpatients, and documents underrecognition of both disorders by resident physicians. Researchers noted TD in 28% of cases and residents only described TD (or symptoms of TD) in 12%. The researcher determined DIP prevalence rate of 26% contrasted with an 11% rate found by residents. Patients with psychotic disorders were more likely than other patients to have researcher-identified TD, whereas DIP (researcher cases) occurred more often in patients with affective diagnoses. Residents tended to miss milder cases of TD, and to miss DIP in younger patients and in patients with affective disorders. Improved teaching and clinical exams are recommended to improve recognition.

Measurement of the biotransfer and time constant of radon from ingested water by human breath analysis.

Forty-one tests were performed on 38 volunteers to measure elimination rates of 222Rn in expired breath. Participants ranged from ages 9 to 85 y, with 16 males and 22 females. The levels of physical activity of the subjects ranged from very inactive to marathon level. Calibration of our flow-through scintillation cell was accomplished using a medical ventilator and 222Rn reservoir for 5-15 L min-1 flow rates. We found a wide range of percent elimination (12-68%) in 30 min. The percent elimination has a mild correlation with the predicted forced expiratory volume in 1 s and with time passed since eating. Our observations of bio-retention half-times range from 17-400 min. The whole-body dose calculations yield a mean of 2.70 +/- 3.43 nGy Bq-1, and the stomach dose calculations yield a mean of 276 +/- 186 nGy Bq-1. These means range beyond those previously reported.

A longitudinal study of correlations among tardive dyskinesia, drug-induced parkinsonism, and psychosis.

Tardive dyskinesia (TD) and drug-induced parkinsonism (DIP) have been hypothesized to reflect opposing states of dopamine (DA) function. In this longitudinal study, 57 psychotic inpatients were rated repeatedly for TD, DIP, and psychosis while receiving neuroleptic medication. Cross-sectional correlations among TD, DIP, and psychosis were weak or nonexistent. Factor and cluster analyses found that 13 patients (23%) were classified into groups characterized by the expected negative correlations. Thus, only partial support was found for the hypothesis that TD and DIP represent opposing states of DA function.

The distal akin osteotomy: a new approach.

The authors describe a modification of the Akin procedure using a distal oblique osteotomy with rigid internal fixation. The anatomic, biomechanic, and physiologic advantages of this modification are discussed. Its proper use and an analysis of results is included.

Erroneous findings in polarized testing caused by plastic prisms.

Certain plastic prisms, when used with polarized tests (eg, stereopsis), can destroy the uniqueness of each eye's view. We investigated this effect by administering the Randot stereotest to 12 visually normal observers. The test was administered monocularly (simulating suppression), under habitual conditions, and through six representative prisms, both fused and monocularly. Whereas stereo thresholds with an anaglyphic random dot stereogram (TNO) were not significantly affected by the prism, fusing through some of the prisms resulted in significantly poorer stereo thresholds and a failure to detect random dot stereopsis (RDS) on the polarized stereotest. Furthermore, some subjects appreciated RDS viewing monocularly through each of the prisms. Thus, it is important to conduct polarized testing through prisms that are known not to interfere with polarization. This optical phenomenon is explained here.

Accurate, TATA box-dependent polymerase III transcription from promoters of the c-myc gene in injected Xenopus oocytes.

We have investigated the factors that permit a gene normally transcribed by RNA polymerase II to be transcribed by RNA polymerase III. It was shown previously that the human c-myc gene could be transcribed in vitro and in Xenopus oocytes by both alpha-amanitin-sensitive and alpha-amanitin-resistant polymerases, probably corresponding to polymerase II and polymerase III. We confirmed this observation in microinjected oocytes and showed that the alpha-amanitin-resistant transcription of c-myc was competed by known polymerase III genes. Polymerase III transcription of c-myc was very inefficient compared to other polymerase III genes, however, and was observed only when large amounts of template DNA were injected. At lower DNA concentrations the gene was transcribed, exclusively by polymerase II. In contrast, the adenovirus major late promoter was not transcribed by polymerase III. The 5' ends of polymerase III RNAs were almost indistinguishable from those of polymerase II RNAs initiating at the P1 and P2 promoters of the human and mouse c-myc genes. Furthermore, point mutations in the TATA box of the human P2 promoter greatly reduced polymerase III activity. At this promoter, therefore, polymerase II and polymerase III recognize a common element, the TATA box, which probably plays an important role in specifying the start site of transcription for both polymerases. We suggest that the highly accurate though inefficient mimicry of polymerase II by polymerase III at the c-myc promoters reflects the common evolutionary origin of these two enzymes.

Risk factors for drug-induced parkinsonism in tardive dyskinesia patients.

Using multivariate statistical analyses, the authors identified risk factors for development of drug-induced parkinsonism (DIP) in 66 tardive dyskinesia (TD) patients. Older age, recent use of neuroleptics, shorter duration of past neuroleptic exposure, and severity of TD were associated with increased risk of DIP. The clinician should devise treatment strategies in anticipation of the occurrence of DIP regardless of the presence or absence of TD, especially in older patients. New models for the pathophysiology of the two disorders are needed.

Covalent Group IV Atomic Clusters.

Atomic clusters containing from two to several hundred atoms offer the possibility of studying the transition from molecules to crystalline solids. The covalent group IV elements carbon, silicon, and germanium are now being examined with this long-range objective. These elements are particularly interesting because of the very different character of their crystalline solids and because they are intermediate between metals and insulators in the nature of their bonding. Small mass-selected atom cluster ions are formed by pulsed laser techniques and identified by time-of-flight methods. Laser photoexcitation is used to study the relative stability of these clusters and their modes of fragmentation. These modes for C(n)(+) clusters, which tend to fragment with a characteristic loss of a neutral C(3), are found to be different from the modes for Si(n)(+) and Ge(n)(+) clusters, which tend to fragment to "magic" clusters such as Si(4)(+), Si(6)(+) and Si(10)(+). These experimental results can be accounted for by recent theoretical calculations of the ground-state structure and stability of small silicon and carbon clusters. Several theoretical approaches give consistent results, showing that small silicon clusters are compact and different from small fragments of the bulk crystal. Calculations show that carbon clusters change from linear structures toward cyclic structures as the cluster size increases, but with significant odd-even differences.

Pharyngo-oesophageal reconstruction by myocutaneous flaps.

With the progressive development of ablative surgery of pharyngolaryngeal cancer in recent decades, the demands for an effective one-stage pharyngo-oesophageal reconstruction have spawned a wide variety of imaginative techniques. The pectoralis major myocutaneous flap has proven to be the most effective for immediate reconstruction of the cervical oesophagus after pharyngolaryngectomy in which the inferior limit of resection is at most 3 cm below the oesophageal orifice. This reconstruction may be circumferential, partially circumferential or only partial in the case of large pharyngostomas.