Exome sequencing in 32 patients with anophthalmia/microphthalmia and developmental eye defects.

Anophthalmia/microphthalmia (A/M) is a genetically heterogeneous birth defect for which the etiology is unknown in more than 50% of patients. We used exome sequencing with the ACE Exome(TM) (Personalis, Inc; 18 cases) and UCSF Genomics Core (21 cases) to sequence 28 patients with A/M and four patients with varied developmental eye defects. In the 28 patients with A/M, we identified de novo mutations in three patients (OTX2, p.(Gln91His), RARB, p.Arg387Cys and GDF6, p.Ala249Glu) and inherited mutations in STRA6 in two patients. In patients with developmental eye defects, a female with cataracts and cardiomyopathy had a de novo COL4A1 mutation, p.(Gly773Arg), expanding the phenotype associated with COL4A1 to include cardiomyopathy. A male with a chorioretinal defect, microcephaly, seizures and sensorineural deafness had two PNPT1 mutations, p.(Ala507Ser) and c.401-1G>A, and we describe eye defects associated with this gene for the first time. Exome sequencing was efficient for identifying mutations in pathogenic genes for which there is no clinical testing available and for identifying cases that expand phenotypic spectra, such as the PNPT1 and COL4A1-associated disorders described here.

PDBe: Protein Data Bank in Europe.

The Protein Data Bank in Europe (PDBe; pdbe.org) is a partner in the Worldwide PDB organization (wwPDB; wwpdb.org) and as such actively involved in managing the single global archive of biomacromolecular structure data, the PDB. In addition, PDBe develops tools, services and resources to make structure-related data more accessible to the biomedical community. Here we describe recently developed, extended or improved services, including an animated structure-presentation widget (PDBportfolio), a widget to graphically display the coverage of any UniProt sequence in the PDB (UniPDB), chemistry- and taxonomy-based PDB-archive browsers (PDBeXplore), and a tool for interactive visualization of NMR structures, corresponding experimental data as well as validation and analysis results (Vivaldi).

Safety, Efficacy and Clinical Applications of Focused Ultrasound-Mediated Blood Brain Barrier Opening in Alzheimer's Disease: A Systematic Review.

Alzheimer's disease is a neurodegenerative disorder marked by cognitive decline and brain pathology involving amyloid plaques and neurofibrillary tangles. Current drug development focuses on disease-modifying therapies, primarily antibodies targeting amyloid or tau. However, the blood-brain barrier (BBB) poses a challenge for drug delivery to the brain. Pre- and early clinical data suggests that Focused Ultrasound (FUS) technology safely enhances BBB permeability without damaging brain tissue, enabling drug delivery. This systematic review discusses the application of FUS to open the BBB for the treatment of Alzheimer's disease (AD). We review the safety, efficacy, and potential biological effects of FUS-mediated BBB opening in AD patients.

Primary and coupled motions after cervical total disc replacement using a compressible six-degree-of-freedom prosthesis.

This study tested the hypotheses that (1) cervical total disc replacement with a compressible, six-degree-of-freedom prosthesis would allow restoration of physiologic range and quality of motion, and (2) the kinematic response would not be adversely affected by variability in prosthesis position in the sagittal plane. Twelve human cadaveric cervical spines were tested. Prostheses were implanted at C5-C6. Range of motion (ROM) was measured in flexion-extension, lateral bending, and axial rotation under ± 1.5 Nm moments. Motion coupling between axial rotation and lateral bending was calculated. Stiffness in the high flexibility zone was evaluated in all three testing modes, while the center of rotation (COR) was calculated using digital video fluoroscopic images in flexion-extension. Implantation in the middle position increased ROM in flexion-extension from 13.5 ± 2.3 to 15.7 ± 3.0° (p < 0.05), decreased axial rotation from 9.9 ± 1.7 to 8.3 ± 1.6° (p < 0.05), and decreased lateral bending from 8.0 ± 2.1 to 4.5 ± 1.1° (p < 0.05). Coupled lateral bending decreased from 0.62 ± 0.16 to 0.39 ± 0.15° for each degree of axial rotation (p < 0.05). Flexion-extension stiffness of the reconstructed segment with the prosthesis in the middle position did not deviate significantly from intact controls, whereas the lateral bending and axial rotation stiffness values were significantly larger than intact. Implanting the prosthesis in the posterior position as compared to the middle position did not significantly affect the ROM, motion coupling, or stiffness of the reconstructed segment; however, the COR location better approximated intact controls with the prosthesis midline located within ± 1 mm of the disc-space midline. Overall, the kinematic response after reconstruction with the compressible, six-degree-of-freedom prosthesis within ± 1 mm of the disc-space midline approximated the intact response in flexion-extension. Clinical studies are needed to understand and interpret the effects of limited restoration of lateral bending and axial rotation motions and motion coupling on clinical outcome.

Evaluation of nature and extent of injuries during Dahihandi festival.

BACKGROUND: Injuries related to the Hindu festival of Dahihandi where a human pyramid is formed and a pot of money kept at a height is broken, celebrated in the state of Maharashtra, have seen a significant rise in the past few years. The human pyramid formed is multi-layered and carries with it a high risk of injury including mortality. AIMS: To evaluate the nature, extent and influencing factors of injuries related to Dahihandi festival. SETTINGS AND DESIGN: We present a retrospective analysis of patients who presented in a tertiary care center with injuries during the Dahihandi festival in the year 2010. MATERIALS AND METHODS: 124 patients' records were evaluated for timing of injury, height of the Dahihandi pyramid, position of the patient in the multi-layered pyramid, mode of pyramid collapse and mechanism of an injury. A binary regression logistic analysis for risk factors was done at 5% significance level. STATISTICAL ANALYSIS: Univariate and multi-variate binary logistic regression of the risk factors for occurrence of a major or minor injury was done using Minitab™ version 16.0 at 5% significance. RESULTS: Out of 139 patients presented to the center, 15 were not involved directly in the formation of pyramid, rest 124 were included in the analysis. A majority of the patients were above 15 years of age [110 (83.6%)]. 46 (37.1%) patients suffered major injuries. There were 39 fractures, 3 cases of chest wall trauma with 10 cases of head injuries and 1 death. More than half of the patients [78 (56.1%)] were injured after 1800 hours. 73 (58.9%) injured participants were part of the pyramid constructed to reach the Dahihandi placed at 30 feet or more above the ground. 72 (51.8%) participants were part of the middle layers of the pyramid. Fall of a participant from upstream layers on the body was the main mechanism of injury, and majority [101 (81.5%)] of the patients suffered injury during descent phase of the pyramid. CONCLUSIONS: There is a considerable risk of serious, life-threatening injuries inherent to human pyramid formation and descent in the Dahihandi festival. Safety guidelines are urgently needed to minimize risk and prevent loss of human life.

Computed tomography-based morphometric analysis of cervical pedicles in Indian population: a pilot study to assess feasibility of transpedicular screw fixation.

BACKGROUND: Cervical transpedicular screw fixation is safe and is probably going to be the gold standard for cervical spine fixation. However, cervical transpedicular screw use in the Asian population can be limited as the transverse diameter in this group of patients may not be adequate to accommodate the 3.5-mm pedicular screw thus injuring the vital structures located in the close proximity of the pedicles. Thus lateral mass fixation remains the mainstay of treatment. The present study evaluated the transverse cervical pedicle diameter of C2-C7 vertebrae in a pilot study in 27 Indian subjects using computed tomography (CT) imaging and evaluated the feasibility of transpedicular screw fixation in them. AIMS: To evaluate the feasibility of transpedicular screw fixation in the Indian population. SETTINGS AND DESIGN: The cervical pedicle diameter size differs between the Asian and non-Asian population. The authors studied the transverse pedicle diameter of the C2-C7 of the cervical spine in the Indian population using CT measurements. This cross-sectional study was carried out at a tertiary care centre for a period of four months from October 2010 to December 2010. MATERIAL AND METHODS: Measurements of cervical pedicles in the subjects were performed on the CT workstation from the CT images taken at 2.5-mm interval. The transverse pedicle diameter was defined as the outermost diameter of the pedicle, taken perpendicular to the axis of the pedicle at the narrowest point and measured in millimeters±0.1 mm. STATISTICAL ANALYSIS: Descriptive statistics was used to represent percentage of transverse diameter of cervical pedicles less than 5 mm in male and female subjects at C2-C7 levels. Since there is no previous study done in India, we initiated the study with sample size of 27 as a pilot study. The statistical analysis was performed using SPSS software. RESULTS: The mean transverse diameters of the cervical pedicles of C2, C3, C4, C5, C6 and C7 in males were 5.3, 5.3, 5.3, 5.6, 5.6 and 6.1 mm respectively and ranged between 5.3 to 6.1 mm. The mean transverse diameters of the cervical pedicles of C2, C3, C4, C5, C6 and C7 in females were 5.1, 4.6, 4.7, 4.7, 5.3 and 5.6 mm respectively and ranged between 4.6 to 5.6 mm. Between 2.1% and 55.7% of pedicles in our male population and between 5.5% and 74.3% pedicles in our female population was smaller than 5.0 mm in transverse diameter and thus cannot have fixation with a 3.5 mm screw using this technique. CONCLUSIONS: We found that the transverse pedicle diameter of cervical pedicles in the Indian subjects is smaller compared to the Western population. Although transpedicular screw fixation has stronger pullout strength compared to lateral mass fixation, its use must be considered carefully and individually. Preoperative CT evaluation is a must before transpedicular fixation in the cervical spine, especially in the Indian female population. As an option 2.7-mm screws can be devised for the Indian population giving a wider safety margin.

A Biomechanical Analysis of Tibial Fixation Methods in Hamstring-Graft Anterior Cruciate Ligament Reconstruction.

Background: There are conflicting results on the biomechanical properties of tibial fixation devices in anterior cruciate ligament reconstruction. The objective of this study is to compare the initial biomechanical properties of tibial fixation in hamstring-graft ACL reconstruction with interference screw, suspension button, and Tape Locking ScrewTM devices. We hypothesized there are no differences in the initial biomechanical properties of these three tibial fixation techniques. Methods: Twenty-one fresh-frozen porcine tibiae were equally divided into three groups of seven tibiae to evaluate the fixation of human hamstring tendon grafts with interference screw, suspension button, or Tape Locking Screw fixation. Using a servohydraulic materials testing system, each graft was subjected to 500 cycles of loading followed by a monotonic failure test. Results: Interference screw fixation demonstrated significantly lower cyclic displacement (1.28 ± 0.73 mm) than the other groups fixated with either a suspension button device (2.54 ± 0.27 mm, p = 0.003) or a Tape Locking Screw (2.32 ± 0.42 mm, p = .009), and a significantly greater cyclic stiffness (212.19 ± 40.30 N/mm) than the Tape Locking Screw (137.64 ± 26.17 N/mm, p = 0.002). The interference screw also demonstrated significantly higher pullout stiffness (166.83 ± 23.22 N/mm) than the suspension button (112.78 ± 24.14 N/mm, P = 0.002) and Tape Locking Screw (109.11 ± 12.91 N/mm, P = 0.0002). Conclusions: Tibial fixation with an interference screw demonstrated superior biomechanical properties for cyclic testing compared to the suspension button and Tape Locking Screw. Load to failure did not differ between groups, and there were no significant biomechanical differences between the suspension button and Tape Locking Screw fixation devices. Clinical Relevance: Despite the initial biomechanical differences, all three fixation devices exhibited mean loads to failure and cyclic displacements below clinically relevant thresholds of failure. These data suggest all three fixation methods are viable options for achieving a functional ACL reconstruction.Level of Evidence: V.

Trigeminal nociceptors express prostaglandin receptors.

Orofacial inflammation is associated with prostaglandin release and the sensitization of nociceptive receptors such as the transient receptor potential subtype V(1) (TRPV(1)). We hypothesized that certain PGE(2) receptor subtypes (EP1-EP4) are co-expressed with TRPV(1) in trigeminal nociceptors and sensitize responses to a TRPV(1) agonist, capsaicin. Accordingly, combined in situ hybridization was performed with immunohistochemistry on rat trigeminal ganglia. We next evaluated the effects of specific EP2 and EP3 agonists (butaprost and sulprostone) in cultured trigeminal ganglia neurons. The results showed that EP2 and EP3 are expressed in trigeminal neurons (58% and 53% of total neurons, respectively) and are co-expressed in TRPV(1)-positive neurons (64% and 67 % of TRPV(1)-positive neurons, respectively). Moreover, most of the cells expressing EP2 or EP3 mRNA were of small to medium diameter (< 30 microm). The application of butaprost and sulprostone triggered neuropeptide exocytosis, and butaprost sensitized capsaicin responses. Analysis of these data, collectively, supports the hypothesis that prostaglandins regulate trigeminal TRPV(1) nociceptors via activation of the EP2 and EP3 receptors.

Biologic therapies for refractory juvenile dermatomyositis: five years of experience of the Childhood Arthritis and Rheumatology Research Alliance in North America.

BACKGROUND: The prognosis of children with juvenile dermatomyositis (JDM) has improved remarkably since the 1960's with the use of corticosteroid and immunosuppressive therapy. Yet there remain a minority of children who have refractory disease. Since 2003 the sporadic use of biologics (genetically-engineered proteins that usually are derived from human genes) for inflammatory myositis has been reported. In 2011-2016 we investigated our collective experience of biologics in JDM through the Childhood Arthritis and Rheumatology Research Alliance (CARRA). METHODS: The JDM biologic study group developed a survey on the CARRA member experience using biologics for Juvenile DM utilizing Delphi consensus methods in 2011-2012. The survey was completed online by the CARRA members interested in JDM in 2012. A second survey was similarly developed that provided more opportunity to describe their experiences with biologics in JDM in detail and was completed by CARRA members in Feb 2013. During three CARRA meetings in 2013-2015, nominal group techniques were used for achieving consensus on the current choices of biologic drugs. A final survey was performed at the 2016 CARRA meeting. RESULTS: One hundred and five of a potential 231 pediatric rheumatologists (42%) responded to the first survey in 2012. Thirty-five of 90 had never used a biologic for Juvenile DM at that time. Fifty-five of 91 (denominators vary) had used biologics for JDM in their practice with 32%, 5%, and 4% using rituximab, etanercept, and infliximab, respectively, and 17% having used more than one of the three drugs. Ten percent used a biologic as monotherapy, 19% a biologic in combination with methotrexate (mtx), 52% a biologic in combination with mtx and corticosteroids, 42% a combination of a biologic, mtx, corticosteroids (steroids), and an immunosuppressive drug, and 43% a combination of a biologic, IVIG and mtx. The results of the second survey supported these findings in considerably more detail with multiple combinations of drugs used with biologics and supported the use of rituximab, abatacept, anti-TNFα drugs, and tocilizumab in that order. One hundred percent recommended that CARRA continue studying biologics for JDM. The CARRA meeting survey in 2016 again supported the study and use of these four biologic drug groups. CONCLUSIONS: Our CARRA JDM biologic work group developed and performed three surveys demonstrating that pediatric rheumatologists in North America have been using multiple biologics for refractory JDM in numerous scenarios from 2011 to 2016. These survey results and our consensus meetings determined our choice of four biologic therapies (rituximab, abatacept, tocilizumab and anti-TNFα drugs) to consider for refractory JDM treatment when indicated and to evaluate for comparative effectiveness and safety in the future. Significance and Innovations This is the first report that provides a substantial clinical experience of a large group of pediatric rheumatologists with biologics for refractory JDM over five years. This experience with biologic therapies for refractory JDM may aid pediatric rheumatologists in the current treatment of these children and form a basis for further clinical research into the comparative effectiveness and safety of biologics for refractory JDM.

The Escherichia coli large ribosomal subunit at 7.5 A resolution.

BACKGROUND: In recent years, the three-dimensional structure of the ribosome has been visualised in different functional states by single-particle cryo-electron microscopy (cryo-EM) at 13-25 A resolution. Even more recently, X-ray crystallography has achieved resolution levels better than 10 A for the ribosomal structures of thermophilic and halophilic organisms. We present here the 7.5 A solution structure of the 50S large subunit of the Escherichia coli ribosome, as determined by cryo-EM and angular reconstitution. RESULTS: The reconstruction reveals a host of new details including the long alpha helix connecting the N- and C-terminal domains of the L9 protein, which is found wrapped like a collar around the base of the L1 stalk. A second L7/L12 dimer is now visible below the classical L7/L12 'stalk', thus revealing the position of the entire L8 complex. Extensive conformational changes occur in the 50S subunit upon 30S binding; for example, the L9 protein moves by some 50 A. Various rRNA stem-loops are found to be involved in subunit binding: helix h38, located in the A-site finger; h69, on the rim of the peptidyl transferase centre cleft; and h34, in the principal interface protrusion. CONCLUSIONS: Single-particle cryo-EM is rapidly evolving towards the resolution levels required for the direct atomic interpretation of the structure of the ribosome. Structural details such as the minor and major grooves in rRNA double helices and alpha helices of the ribosomal proteins can already be visualised directly in cryo-EM reconstructions of ribosomes frozen in different functional states.

Databases and Archiving for CryoEM.

CryoEM in structural biology is currently served by three public archives-EMDB for 3DEM reconstructions, PDB for models built from 3DEM reconstructions, and EMPIAR for the raw 2D image data used to obtain the 3DEM reconstructions. These archives play a vital role for both the structural community and the wider biological community in making the data accessible so that results may be reused, reassessed, and integrated with other structural and bioinformatics resources. The important role of the archives is underpinned by the fact that many journals mandate the deposition of data to PDB and EMDB on publication. The field is currently undergoing transformative changes where on the one hand high-resolution structures are becoming a routine occurrence while on the other hand electron tomography is enabling the study of macromolecules in the cellular context. Concomitantly the archives are evolving to best serve their stakeholder communities. In this chapter, we describe the current state of the archives, resources available for depositing, accessing, searching, visualizing and validating data, on-going community-wide initiatives and opportunities, and challenges for the future.

Preparation and electrochemical behavior of gramicidin-bipolar lipid monolayer membranes supported on gold electrodes.

Gramicidin-containing synthetic bolalipid membranes comprised of 2,2'-di-O-decyl-3,3'-O-1,20-eicosanyl-bis-rac-glycero-1,1'-diphosphocholine (C20BAS) have been synthesized and supported on gold electrodes. Supported membranes were prepared by first depositing a partial bolalipid layer on the electrode using a thioctic acid-modified bolalipid (1'-O-omega-thioctamidetetraethylene glycol-2,2'-di-O-decyl-3,3'-di-O-1,20-eicosanyl-bis-rac-glycero-1-phosphate, SSC20BAS) as an anchoring group, followed by a vesicle fusion step using either pure C20BAS or gramicidin-containing C20BAS (C20BAS-GA) vesicles. The latter configuration was designed to immobilize single, continuously-on channels of gramicidin in the C20BAS membrane. Vesicle deposition to form supported bolalipid monolayer membranes was monitored by impedance spectroscopy and cyclic voltammetry. Impedances were observed to increase with vesicle deposition time. Pretreatment of the impedance electrode with SSC20BAS accelerated the supported monolayer membrane deposition rate. Impedances decreased in a gramicidin concentration-dependent manner when gramicidin was incorporated into the C20BAS membrane. These supported bolalipid membranes are also surprisingly inert to organic solvent exposure (CH(3)CH(2)OH;CH(2)Cl(2)), suggesting that they may serve as robust host matrices for integral membrane protein-based sensors.

False positive head-up tilt: hemodynamic and neurohumoral profile.

OBJECTIVES: This study examined differences in mechanisms of head-up tilt (HUT)-induced syncope between normal controls and patients with neurocardiogenic syncope. BACKGROUND: A variable proportion of normal individuals experience syncope during HUT. Differences in the mechanisms of HUT-mediated syncope between this group and patients with neurocardiogenic syncope have not been elucidated. METHODS: A 30-min 80 degrees HUT was performed in eight HUT-negative volunteers (Group I), eight HUT-positive volunteers (Group II) and 15 patients with neurocardiogenic syncope. Heart rate and blood pressure (BP) were monitored continuously. Epinephrine and norepinephrine plasma levels, as well as left ventricular dimensions and contractility determined by echocardiography, were measured at baseline and at regular intervals during the test. RESULTS: The main findings of this study were the following: 1) All parameters were similar at baseline in the three groups; and 2) During tilt: a) the time to syncope was shorter in Group III than in group II (9.5 +/- 3 vs. 17 +/- 3 min p < 0.05); b) there was an immediate, persisting drop in mean BP in Group III; c) the decrease rate of left ventricular end-diastolic dimensions was greater in Group III than in Group II or Group I (-1.76 +/- 0.42 vs. -0.87 +/- 0.35 and -0.67 +/- 0.29 mm/min, respectively, p < 0.05); d) the leftventricular shortening fraction was greater in Group III than in the other two groups (39 +/- 1 vs. 34 +/- 1 and 32 +/- 1%, respectively, p < 0.05); and e) although the norepinephrine level remained comparable among the groups, there was a significantly higher peak epinephrine level in Group III than in Group II and Group I (112.3 +/- 34 vs. 77.6 +/- 10 and 65 +/- 12 pg/ml, p < 0.05). CONCLUSIONS: Mechanisms of syncope during HUT appeared to be different in normal volunteers and patients with neurocardiogenic syncope. In the latter, there was evidence of an impaired vascular resistance response from the beginning of the orthostatic challenge. Furthermore, in the patients there was more rapid peripheral blood pooling, as indicated by the echocardiographic measurements of left ventricular end-diastolic changes, leading to more precocious symptoms. In syncopal patients, the higher level of plasma epinephrine probably mediated the increased cardiac contractility and possibly contributed to the impaired vasoconstrictive response.

A purification strategy for inositol 1,4,5-trisphosphate 3-kinase from rat liver based upon heparin interaction chromatography.

Rat liver inositol 1,4,5-trisphosphate [Ins (1,4,5)P3] 3-kinase was purified in high yield by a three-step procedure reliant upon chromatography on heparin and calmodulin agarose. Purified enzyme was stable in the presence of the detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS) (0.1-0.5%) and the sulphydryl reducing reagent dithiothreitol (DTT). The purified enzyme was activated 2-3-fold by Ca2+ (1 microM) in the presence of calmodulin. Pyrophosphate and heparin were identified as inhibitors of the enzyme.

Larvicidal activity of the fungus Aphanomyces (oomycetes : Saprolegniales) against Culex quinquefasciatus.

Several species of fungi are currently being considered for operational use in the microbial control of mosquito larvae. The oomycetous fungi are the prominent ones amongst them because of their ability to complete life cycle in water. During our studies on zoosporic fungi from riverine waters of Mula and Mutha flowing through Pune City, Maharashtra, India, Aphanomyces laevis de Bary (Oomycetes: Saprolegniales) was isolated from polluted waters. After critical observations it was found to be mosquito larvicidal. Sporulating hemp seed cultures when inoculated under laboratory conditions revealed that it causes 80%/ mortality after seven days to Culex quinquefasciatus larvae. Laboratory assays were conducted to determine the effects of water quality on the ability of the isolate to infect mosquito larvae in varying degrees of pollution levels. In all the experiments, a non sexual strain of Aphanomyces (zoospores) was found to be the pathogenic agent for the Culex larvae.

Ablation surgery for atrial fibrillation: "freeze it or buzz it; just do it and cure it".

Patients in normal sinus rhythm have lesser stroke rate, better functional class and quality of life than those in atrial fibrillation. Adding a surgical procedure to cure atrial fibrillation in patients needing correction of structural heart disease has been shown to be a safe option, which benefits the majority in restoration of sinus rhythm. Age is no bar to implement this option. The same does not hold true for lone atrial fibrillation. The affirm trial has shown that there is need for improved treatment strategies for patients in atrial fibrillation, although young patients were not represented in sizable proportion. There is need to develop curative treatment for patients with lone atrial fibrillation. And there are technological advances in the form of ablative energy sources and hardware for applying these with minimal invasion. "Between tomorrow's dream and yesterday's regret is today's opportunity". Let's make the best of it!

A study of the precision of confocal, ratiometric, Fura-2-based [Ca2+] measurements.

The precision with which quantitative confocal ratiometric Fura-2-based calcium measurements can be performed has been investigated. A standard confocal scanning laser microscope system has been modified so as to enable excitation with the 351 nm and 364 nm lines of the UV argon laser and simultaneous separation of the fluorescence emanating from the two different excitation wavelengths. Experiments have been performed on living cells and mainly on Fura-2 containing calibrated Ca2+ buffer solutions in order to study the variation of the precision with various experimental parameters in a controlled manner. Furthermore, expressions relating the precision of the calcium determination to various measured parameters are derived. These expressions have been used in conjunction with published excitation intensity spectra for Fura-2 in order to verify the experimental results and to analyse the variation of parameters that could not be experimentally tested. The results show that a photon number of the order of 10,000 per pixel, at maximum Ca2+ concentration and 351 nm excitation, is required in order to reduce the relative error at all concentrations to less than 30%. The commercially available laser wavelength combinations, 325 nm/380 nm and 334 nm/380 nm, were found to be the best as concerns the precision of the method.

Attenuation of frontal asymmetry in pediatric posttraumatic stress disorder.

BACKGROUND: Volumetric imaging research has shown abnormal brain morphology in adults with posttraumatic stress disorder (PTSD) when compared with matched control subjects. In this article, we present brain imaging findings from a study of children with PTSD symptoms. METHODS: Twenty-four children between the ages of 7 and 14 with a history of trauma and PTSD symptoms were assessed with the Clinician-Administered PTSD Scale for Children and Adolescents (CAPS-CA). The sample underwent magnetic resonance imaging in a 1.5 T scanner. Brain images were analyzed by raters blind to diagnostic status using well-standardized methods, and images were compared with age- and gender-matched healthy control subjects. RESULTS: The clinical group demonstrated attenuation of frontal lobe asymmetry and smaller total brain and cerebral volumes when compared with the control group. There were no statistically significant differences in hippocampal volume between clinical and control subjects. CONCLUSIONS: Frontal lobe abnormalities may occur as a result of PTSD in children or, alternatively, be a risk factor for the development of the syndrome in this age group. The implications of the findings and their consistency with previous research are discussed.

Brain morphology in Klinefelter syndrome: extra X chromosome and testosterone supplementation.

OBJECTIVE: This study focuses on variation in brain morphology associated with supernumerary X chromosome and Klinefelter syndrome (KS). Using an unselected birth cohort of KS subjects and high-resolution MRI, the authors investigated the neuroanatomic consequences of the 47,XXY karyotype in the presence and absence of exogenous testosterone supplementation. METHODS: Regional brain volumes were measured in 10 subjects with KS and 10 age-matched control men. Five of the KS subjects had received testosterone supplementation since puberty (KS+T) and five had not (KS-T). RESULTS: KS subjects showed significant (p < 0.01) reduction in left temporal lobe gray matter volumes compared with normal control subjects. Differences in left temporal gray volumes were also significant between the KS+T and KS-T groups (p < 0.01). Verbal fluency scores were significantly different between the KS+T and KS-T groups as well. CONCLUSION: Supernumerary X chromosome material in men is associated with a reduction in left temporal lobe gray matter, a finding that is consistent with the verbal and language deficits associated with KS. Also, relative preservation of gray matter in the left temporal region is associated with exposure to exogenous androgen during development. A history of testosterone supplementation also appears to be associated with increased verbal fluency scores in KS patients.

Heterozygous oim mice exhibit a mild form of osteogenesis imperfecta.

The oim strain of mice is one of several rodent models that exhibit an osteogenesis imperfecta (OI) phenotype. These mice have a mutation in the gene encoding alpha-2 chain of type I procollagen that prevents proper assembly of this propeptide with alpha-1 propeptides. Homozygous oim mice experience multiple bone fractures under standard laboratory animal housing conditions and are representative of moderate to severe forms of OI. Because fractures are not typically experienced by heterozygous oim mice, they have not been studied extensively. The present studies show that the organization of cortical bone is deficient in heterozygotes, exhibiting a morphology intermediate to specimens from homozygotes and wild-type mice. The biomechanical properties of femurs isolated from heterozygous oim mice are also intermediate to homozygotes and wild-type mice when tested in four-point bending. Although it is not possible to distinguish visually between heterozygous oim and wild-type mice, the quality and biomechanical properties of bone in heterozygotes is significantly reduced by twelve weeks of age. Heterozygous oim mice are useful as a model for a mild form of OI.