Age-dependent change in the hyaluronic acid content of the human chorioretinal complex.
Hyaluronic acid (HA) has a key role in the structure and organization of the extracellular matrix. We sought to identify the distribution of HA in human eye tissue with regard to age using a biotinylated HA-binding protein. Fetal and adult (from donors ranging from 28 to 94 years of age) eye tissues were fixed less than 24 hours post mortem and embedded in JB-4 medium (Polysciences, Warrington, Pa). Sections of 2-microns thickness were used. Control sections were pretreated either with Streptomyces hyaluronidase or HA-binding protein inactivated by HA. The binding of the protein to HA was detected with avidinbiotin alkaline phosphatase and developed by incubation with naphthol as-mx phosphate and Texas Red Salt (Pierce, Rockford, Ill). Specific staining for HA was observed in fetal eyes in the choroid, Bruch's membrane, sclera, retinal pigment epithelium, and developing retina from the vitreoretinal interface to the inner plexiform layer. Specific staining decreased with age in the choroid, retinal pigment epithelium, and Bruch's membrane. Hyaluronic acid-specific staining was undetectable in tissues from donors over 50 years of age. The localization of HA in the chorioretinal complex and its disappearance after the fifth decade of life may play a role in aging and age-related retinal disorders.


Upper extremity venous aneurysm as a source of pulmonary emboli.
Spontaneous venous aneurysms of the upper extremities and neck are rare and typically asymptomatic. We present the first reported case of a symptomatic, primary upper extremity venous aneurysm in a patient who initially presented with pulmonary emboli. A 22-year-old patient was admitted with chest pain, dyspnea, and a right axillary mass. Computed tomography revealed diffuse, bilateral pulmonary emboli in addition to a thrombosed axillary vein. Venography confirmed the diagnosis, and also revealed a subclavian vein stenosis at the crossing of the first rib. Pharmacomechanical thrombolysis, catheter-directed thrombolysis, and venoplasty were performed with adequate flow restoration, also revealing the presence of a previously thrombosed proximal brachial/distal axillary venous aneurysm. Hematologic testing showed a positive and persistent lupus anticoagulant. The patient declined surgical reconstruction and opted for long-term anticoagulation. At 24 months, the patient continued to remain symptom-free.


FGFR3-targeted mAb therapy for bladder cancer and multiple myeloma.
Gain-of-function mutations in FGF receptor 3 (FGFR3) have been implicated in severe skeletal dysplasias and in a variety of cancers. In their study in this issue of the JCI, Qing et al. used specific shRNA probes to demonstrate that FGFR3 functions as an important driver of bladder carcinoma cell proliferation (see the related article beginning on page 1216). A unique anti-FGFR3 mAb was shown to exhibit antitumor activity in human bladder carcinoma cells in vitro and in mouse bladder cancer or multiple myeloma xenograft tumor models bearing either wild-type or mutant FGFR3. These results suggest that clinical development of anti-FGFR3 mAbs should be considered for targeted therapy of cancer and other diseases.


Impact of tumor vascularity on responsiveness to antiangiogenesis in a prostate cancer stem cell-derived tumor model.
Drugs that target the tumor vasculature and inhibit angiogenesis are widely used for cancer treatment. Individual tumors show large differences in vascularity, but it is uncertain how these differences affect responsiveness to antiangiogenesis. We investigated this question using two closely related prostate cancer models that differ markedly in tumor vascularity: PC3, which has very low vascularity, and the PC3-derived cancer stem-like cell holoclone PC3/2G7, which forms tumors with high microvessel density, high tumor blood flow, and low hypoxia compared with parental PC3 tumors. Three angiogenesis inhibitors (axitinib, sorafenib, and DC101) all induced significantly greater decreases in tumor blood flow and microvessel density in PC3/2G7 tumors compared with PC3 tumors, as well as significantly greater decreases in tumor cell proliferation and cell viability and a greater increase in apoptosis. The increased sensitivity of PC3/2G7 tumors to antiangiogenesis indicates they are less tolerant of low vascularity and suggests they become addicted to their oxygen- and nutrient-rich environment. PC3/2G7 tumors showed strong upregulation of the proangiogenic factors chemokine ligand 2 (CCL2) and VEGFA compared with PC3 tumors, which may contribute to their increased vascularity, and they have significantly lower endothelial cell pericyte coverage, which may contribute to their greater sensitivity to antiangiogenesis. Interestingly, high levels of VEGF receptor-2 were expressed on PC3 but not PC3/2G7 tumor cells, which may contribute to the growth static response of PC3 tumors to VEGF-targeted antiangiogenesis. Finally, prolonged antiangiogenic treatment led to resumption of PC3/2G7 tumor growth and neovascularization, indicating these cancer stem-like cell-derived tumors can adapt and escape from antiangiogenesis.


Minimally invasive evacuation of parenchymal and ventricular hemorrhage using the Apollo system with simultaneous neuronavigation, neuroendoscopy and active monitoring with cone beam CT.
The Apollo system is a low profile irrigation-aspiration system which can be used for the evacuation of intracranial hemorrhage. We demonstrate the feasibility of using Apollo to evacuate intracranial hemorrhage in a series of three patients with combined neuronavigation, neuroendoscopy, and cone beam CT (CB-CT). Access to the hematoma was planned using neuronavigation software. Parietal (n=2) or frontal (1) burr holes were created and a 19 F endoscopic sheath was placed under neuronavigation guidance into the distal aspect of the hematoma along its longest accessible axis. The 2.6 mm Apollo wand was then directed through the working channel of a neuroendoscope and used to aspirate the blood products under direct visualization, working from distal to proximal. After a pass through the hematoma, the sheath, neuroendoscope, and Apollo system were removed. CB-CT was then used to evaluate for residual hematoma. When required, the CB-CT data could then be directly uploaded into the neuronavigation system and a new trajectory planned to approach the residual hematoma. Three patients with parenchymal (n=2) and mixed parenchymal-intraventricular (n=1) hematomas underwent minimally invasive evacuation with the Apollo system. The isolated parenchymal hematomas measured 93.4 and 15.6 mL and were reduced to 11.2 (two passes) and 0.9 mL (single pass), respectively. The entire parenchymal component of the mixed hemorrhage was evacuated, as was the intraventricular component within the right frontal horn (single pass). No complications were experienced. All patients showed clinical improvement after the procedure. The average presenting National Institutes of Health Stroke Scale was 19.0, which had improved to 5.7 within an average of 4.7 days after the procedure. The Apollo system can be used within the neuroangiography suite for the minimally invasive evacuation of intracranial hemorrhage using simultaneous neuronavigation for planning and intraprocedural guidance, direct visualization with neuroendoscopy, and real time monitoring of progress with CB-CT.


Analysis of the albino-locus region of the mouse. I. Origin and viability.
Numerous specific-locus experiments designed to test the mutagenic effect of external radiation have yielded, in over 3,600,000 animals observed, altogether 119 presumed mutations involving the c locus. Of these, 55 were viable and albino (cav), 13 were viable and of various intermediate pigment types (cxv), four were subvital (cas and cxs), seven were neonatally lethal albinos (cal), 28 prenatally lethal albinos (cal); 12 died untested. All of the prenatally lethal and at least one of the neonatally lethal c-locus mutations (cal classes) are probably deficiencies that we have analyzed extensively in other experiments. Since absence of the locus mimics albino in phenotype, the intermediates (cxv and cxs groups) probably resulted from intragenic changes. The class of viable albino mutants (cav) might include, in addition to intragenic changes, some extremely small deficiencies. --The effects on viability of c-locus lethals (cal's) in heterozygous condition are not drastic enough to be perceived in stocks of mixed genetic background except in the case of the two longest known deficiencies and a few others. --Analysis of the relation between radiation treatment and type of c-locus mutants obtained shows that the relative frequency of viable mutations, for each germ-cell type, is greater for low-LET than for neutron irradiation; however, the difference for any individual cell type is not significant. The majority (66.7%) of mutations derived from X- or gamma-ray irradiated spermatogonia are viable, and the proportion of "intermediates" among these viables is similar to that among presumed spontaneous c-locus mutations. No significant dose-rate effect on the proportion of lethals could be demonstrated within the set of mutants induced by low-LET irradiation of spermatogonia. Although sets from other germ-cell stages are too small for statistical tests, the results for oocytes are similar, as far as they go. Furthermore, most of the c-locus mutations induced in spermatogonia, even by high-dose-rate X-ray or gamma irradiation, are of a type most likely to result from single-tract events (62% cxv, cxs, and cav; plus 16% presumed deficiencies not involving the closest marker). These results support the view that most of the reduction in mutation frequency at low dose rates is not due to a change in relative proportion of two-track and one-track ionizing events.


Mitochondria as the Target of Hepatotoxicity and Drug-Induced Liver Injury: Molecular Mechanisms and Detection Methods.
One of the major mechanisms of drug-induced liver injury includes mitochondrial perturbation and dysfunction. This is not a surprise, given that mitochondria are essential organelles in most cells, which are responsible for energy homeostasis and the regulation of cellular metabolism. Drug-induced mitochondrial dysfunction can be influenced by various factors and conditions, such as genetic predisposition, the presence of metabolic disorders and obesity, viral infections, as well as drugs. Despite the fact that many methods have been developed for studying mitochondrial function, there is still a need for advanced and integrative models and approaches more closely resembling liver physiology, which would take into account predisposing factors. This could reduce the costs of drug development by the early prediction of potential mitochondrial toxicity during pre-clinical tests and, especially, prevent serious complications observed in clinical settings.


Frailty in Patients With Trauma Who Are Critically Ill: A Prospective Observational Study to Determine Feasibility, Concordance, and Construct and Predictive Validity of 2 Frailty Measures.
As the older population increases, more older people are exposed to trauma. Frailty can be used to highlight patients at risk of a poorer outcome. The objectives of this study were to compare 2 frailty measures with regard to concordance, floor and ceiling effects, and construct and predictive validity and to determine which is more valid and clinically applicable in a critically ill trauma population. This was a prospective observational study. Patients were included if admitted to an intensive care unit (ICU) under a trauma medical unit and ≥ 50 years old. Frailty was determined using 2 frailty measures, the Frailty Phenotype (FP) and Clinical Frailty Scale (CFS). One hundred people were enrolled; their mean age was 69.2 years (SD = 10.4) and 81% had major trauma (as determined with the Injury Severity Score). Frailty was identified with the FP in 22 participants and with the CFS in 13 participants. The 2 frailty measures had an excellent correlation (Spearman rank correlation coefficient = 0.77; 95% confidence interval = 0.66-0.85). Both the FP and the CFS had large floor effects but no ceiling effects. The FP and CFS showed construct validity, with frailty being significantly associated with increasing age, requiring an aid to mobilize, and more falls and hospital admissions. Frailty on the FP was predictive of ICU and hospital mortality, whereas frailty on the CFS was predictive of hospital mortality. The limitations of this study include the use of a single site, small sample size, and collection of frailty measures retrospectively. Measuring frailty in a trauma ICU population was feasible, with excellent correlation between the 2 frailty measures. Both showed aspects of construct and predictive validity; however, the FP identified frailty in more participants and was associated with more comorbidities and higher mortality at ICU discharge. Therefore, the FP might be more clinically relevant in this population.


Identification of a missense variant in CLDN2 in obstructive azoospermia.
Obstructive azoospermia (OA), defined as an obstruction in any region of the male genital tract, accounts for 40% of all azoospermia cases. Of all OA cases, ~30% are thought to have a genetic origin, however, hitherto, the underlying genetic etiology of the majority of these cases remain unknown. To address this, we took a family-based whole-exome sequencing approach to identify causal variants of OA in a multiplex family with epidydimal obstruction. A novel gain-of-function missense variant in CLDN2 (c.481G>C; p.Gly161Arg) was found to co-segregate with the phenotype, consistent with the X-linked inheritance pattern observed in the pedigree. To assess the pathogenicity of this variant, the wild and mutant protein structures were modeled and their potential for strand formation in multimeric form was assessed and compared. The results showed that dimeric and tetrameric arrangements of Claudin-2 were not only reduced, but were also significantly altered by this single residue change. We, therefore, envisage that this amino acid change likely forms a polymeric discontinuous strand, which may lead to the disruption of tight junctions among epithelial cells. This missense variant is thus likely to be responsible for the disruption of the blood-epididymis barrier, causing dislodged epithelial cells to clog the genital tract, hence causing OA. This study not only sheds light on the underlying pathobiology of OA, but also provides a basis for more efficient diagnosis in the clinical setting.


PI3K/Akt/mTOR signaling & its regulator tumour suppressor genes PTEN & LKB1 in human uterine leiomyomas.
Despite their high occurrence and associated significant level of morbidity manifesting as spectrum of clinical symptoms, the pathogenesis of uterine leiomyomas (ULs) remains unclear. We investigated expression profile of tumour suppressor genes PTEN (phosphatase and tensin homolog deleted on chromosome ten) and LKB1 (liver kinase B1), and key signaling components of P13K (phosphatidylinositol 3-kinase)/Akt (protein kinase B)/mTOR (mammalian target of rapamycin) pathway in leiomyomas and adjacent normal myometrium in women of reproductive age, to explore the possibility of targeting this pathway for future therapeutic implications. Real time PCR (qPCR) was used to quantify relative gene expression levels of PTEN, Akt1, Akt2, mTOR, LKB1 and VEGFA (vascular endothelial growth factor A) in leiomyoma as compared to adjacent normal myometrium. Immunohistochemistry was subsequently performed to analyze expression of PTEN, phospho-Akt, phospho-mTOR, phospho-S6, LKB1 and VEGFA in leiomyoma and adjacent normal myometrium. Significant upregulation of PTEN (2.52 fold; P=0.03) and LKB1 (3.93 fold; P0.01), and downregulation of VEGFA (2.95 fold; P=0.01) genes were observed in leiomyoma as compared to normal myometrium. Transcript levels of Akt1, Akt2 and mTOR did not vary significantly between leiomyoma and myometrium. An increased immunoexpression of PTEN (P=0.015) and LKB1 (P<0.001) and decreased expression of VEGFA (P=0.01) was observed in leiomyoma as compared to myometrium. Immunostaining for activated (phosphorylated) Akt, mTOR and S6 was absent or low in majority of leiomyoma and myometrium. Upregulation of PTEN and LKB1 in concert with negative or low levels of activated Akt, mTOR and S6 indicates that PI3K/Akt/mTOR pathway may not play a significant role in pathogenesis of leiomyoma.


The complex between a four-way DNA junction and T7 endonuclease I.
The junction-resolving enzyme endonuclease I is selective for the structure of the DNA four-way (Holliday) junction. The enzyme binds to a four-way junction in two possible orientations, with a 4:1 ratio, opening the DNA structure at the centre and changing the global structure into a 90 degrees cross of approximately coaxial helices. The nuclease cleaves the continuous strands of the junction in each orientation. Binding leads to pronounced regions of protection of the DNA against hydroxyl radical attack. Using all this information together with the known structure of the enzyme and the structure of the BglI-DNA complex, we have constructed a model of the complex of endonuclease I and a DNA junction. This shows how the enzyme is selective for the structure of a four-way junction, such that both continuous strands can be accommodated into the two active sites so that a productive resolution event is possible.


Retrospective assessment of the dismasking flap procedure as a craniofacial approach.
In 1997 the authors presented the case of a 3-year-old girl who underwent complete resection of a huge tumor via a new technique using a dismasking flap. Since that report, 14 patients have undergone surgery using this technique. There have been few reports on long-term follow-up after a craniofacial surgical approach and reconstruction of a huge tissue defect. The authors report details of this procedure based on these cases, including long-term follow-up in the original case. The dismasking flap is a cranioorbitofacial degloving method that uses a circumpalpebral approach with or without piriform margin incisions and presents a wide surgical field under direct vision. Fourteen patients have undergone surgery using the dismasking flap technique. These patients had multiple craniofacial bone fractures (3 cases) and malignant or premalignant craniofacial tumors (11 cases). Patients ranged in age from 3 to 62 years old. The longest follow-up period was 15 years. The results of these cases are reported regarding changes in the facial bones and soft tissues with growth, the various pathologies involved, and complications. Complications using this approach were lagophthalmos with ectropion of the lower eyelid, disturbance of the superior orbital nerve, disturbance of the inferior orbital nerve, maxillary hypoplasia, and blepharoptosis. In all affected patients these complications were almost always temporary. In 2 of the 14 cases, a repair operation for lagophthalmos was necessary, while others showed spontaneous improvement approximately 6 months to a year after the procedure. In the case with the longest follow-up duration, the patient's nose is asymmetrical, especially the alar portion, due to hypoplasia of the grafted bone and/or the fixation with titanium miniplates. This flap is very useful for en bloc resection of huge skull base tumors, multiple craniofacial bone fractures, and as a lateral approach to a deep portion of the middle cranial base. Careful attention is necessary, however, because one may encounter unexpected complications if one does not ensure adequate protection of the perioperative flap. Meticulous postoperative management is also essential.


Virus diagnostics on microarrays.
Whereas the majority of microarray applications still deal with expression analysis for gathering information about levels of gene products at certain cell states, other approaches simply ask the question whether particular genes, which are usually indicative for particular microorganisms and pathogens, are present in a sample or not. Investigations that are more detailed try to evaluate the presence of particular subtypes of a given pathogen. The combination of microarray technology and virus diagnostics promises to generate an ideal platform for fast, sensitive, specific, and parallelized virus diagnostics. Performing virus diagnostics on microarrays, however, requires other basic techniques to be optimized. This is necessary in order to obtain unambiguous and reproducible results, which are compatible with the needs for clinical routine. Parameters that have to be considered include supports, coupling chemistry, chemical oligonucleotide synthesis, signal enhancement strategies, and optimal coordination of PCR reactions, hybridizations, and signal detection, as well as interpretation strategies. Finally, considerations should be given to economic aspects, one chip-one patient strategies and low integrated arrays as a custom-tailored way to fast and accurate diagnostic tools.


Effects of polymorphisms in CAPN1 and CAST genes on meat tenderness of Chinese Simmental cattle.
Considerable evidence has demonstrated that the μ -calpain (CAPN1) gene and its inhibitor calpastatin (CAST) gene are major factors affecting meat quality. Marker-assisted selection (MAS) has been widely used to improve beef quality traits. Therefore, the objective of the present study was to investigate the single nucleotide polymorphisms (SNPs) of bovine CAPN1 and CAST genes using 367 animals representing the four main Chinese cattle breeds and to explore the effects of these SNPs on meat quality traits. Two SNPs within CAPN1 and one SNP in CAST were successfully identified in cattle. Genetic diversity analyses suggested that most SNPs in the four breeds exhibited a moderate genetic diversity. Moreover, associations between individual markers and meat quality traits were analyzed in Chinese Simmental cattle. The CAPN1 4558 A > G locus was found to be significantly associated with shear force value (SFV) and marbling score (BMS), and CAPN1 4684 C > T exerted a significant effect on SFV, while the CAST genotype was not significantly associated with any of the measured traits. SFV, commonly used to measure meat tenderness, represents an important quality trait as it contributes to the flavor of cooked meat. This work confirms the effect of CAPN1 on beef tenderness and lays an important foundation for future cattle breeding.


Melanocyte-stimulating activity in fetal, newborn and adult guinea-pigs: studies of the pituitary, blood plasma and amniotic fluid.
Melanocyte-stimulating activity in the pituitary and plasma of fetal, newborn and adult guinea-pigs, and in amniotic fluid, was bioassayed on isolated Anolis skin. In the fetuses, levels in the posterior, anterior and whole pituitary increased exponentially between 39 and 70 days of gestation; in all cases, the increases were close to 35-fold in terms of ng/lobe, and 6-fold in ng/mg of lyophilized tissue. The increases were due to growth, and to an increased accumulation within the tissues. Ratios of activity in the posterior pituitary: anterior pituitary: hypothalamus approximated 200:30:1 (ng/mg of dry tissue). Fetal plasma levels showed some scatter; however, there were no trends with age, and the average value (10.0 +/- 1.6 ng/ml) was close to that of the mothers (10.6 +/- 2.6 ng/ml). Plasma activity appeared to fall markedly around birth; newborn pups, 2-6 hours after birth, showed levels 65% below those of the fetuses, and also below those of the adults (significant; P less than 0.05). At the same time the pituitary levels increased; this suggested a shut-down in release. The possible implications of this in the birth processes are discussed. Amniotic fluid showed only low, inconsistent activity, below 1% of plasma levels.


Rotavirus serotypes and electropherotypes identified among hospitalised children in São Luís, Maranhão, Brazil.
During June 1997-June 1999 rotavirus infection was screened in infants aged up to 2 years and hospitalised with acute diarrhoea in São Luís, Northeastern Brazil. Altogether, 128 stool samples were collected from diarrhoeic patients and additional 122 faecal specimens from age and temporal matched inpatients without diarrhoea were obtained; rotavirus positivity rates for these groups were 32.0% (41/128) and 9.8% (12/122), respectively (p < 0.001). Both electropherotyping and serotyping could be performed in 42 (79.2%) of the 53 rotavirus-positive stool samples. Long and short electropherotypes were detected at similar rates--38.1% and 40.5% of specimens, respectively. Overall, a G serotype could be assigned for 35 (83.3%) of specimens, the majority of them (66.7%) bearing G1-serotype specificity. Taking both electropherotypes and serotypes together, G1 rotavirus strains displaying long and short RNA patterns accounted for 30.9% and 19.0% of tested specimens, respectively; all G2 strains had short electropherotype. Rotavirus gastroenteritis was detected year-round and, in 1998, the incidence rates tended to be higher during the second semester than in the first semester: 45.2% and 26.1% (p = 0.13), respectively. Rotavirus infections peaked at the second semester of life with frequencies of 30.1% and 13.5% for diarrhoeic children and controls, respectively. While the six rotavirus strains bearing G2-type specificity were circulating throughout the whole study period, G1 serotypes (n = 27) emerged as from June 1998 onwards, 20 (74.1%) of which clustering in 1998. These data underscore the importance of rotaviruses in the aetiology of severe infantile gastroenteritis in Northeastern Brazil and sustain the concept that a future vaccine should confer protection against more than one serotype.


[A mathematical model of the status of mitochondrial DNA for assessing the effectiveness of energy biosynthesis in mammalian cells].
A concept is advanced according to which the processes of ageing, non-specific pathology and malignant transformation are caused by mutations of mitochondrial DNA which controls protein synthesis of the oxidation phosphorylation system. A mathematical model is plotted for describing the efficiency of the system of energy supply of mammalian cells as a function of accumulation of mtDNA mutations. It is shown that at a definite degree of heterogeneity of mitochondria population malignant transformation proceeds. Explanations are presented of the existence of a latent period between the effect of cancerogenic agent and occurrence of tumour, mechanism of the realization of remote effects of chronic action of low intensity agents (these explanations are absent in modern theories of cancerogenesis and quantitative radiobiology).


[Comparative genetics of yeasts. XXII. The determination of alpha-methylglucoside fermentation by the maltose genes MAL6c2 and malx in the offspring of Saccharomyces cerevisiae N. C. Y. C. 74 strain].
In offsprings of N.C.Y.C. 74, maltose regulatory constitutive MAL6C2 mutation controls alpha-methylglucoside (alpha-mgl) fermentation in the presence of MALx. MAL6C2 MALx system described by ten Berge et al, is analogous in function (polymeric interaction) to, at least, one MGLa gene from the system of complementary alpha-mgl genes MGLa MGLb MGLc identified by ten Berge. Suppressor malx mutation inhibits both the maltose and alpha-mgl activity of MAL6C2 allele. A brief review on participation of maltose genes in alpha-mgl fermentation is presented.


The thyroid risk score (TRS) for nodules with indeterminate cytology.
Cytology is the gold standard method for the differential diagnosis of thyroid nodules, though 25-30% of them are classified as indeterminate. We aimed to set up a 'thyroid risk score' (TRS) to increase the diagnostic accuracy in these cases. We prospectively tested 135 indeterminate thyroid nodules. The pre-surgical TRS derived from the sum of the scores assigned at cytology, EU-TIRADS classification, nodule measurement, and molecular characterization, which was done by our PTC-MA assay, a customized array able to cost-effectively evaluate 24 different genetic alterations including point mutations and gene fusions. The risk of malignancy (ROM) increased paralleling the score: in the category >4 and ≤ 6 (low suspicion), >6 ≤ 8 (intermediate suspicion), and >8 (high suspicion); ROM was 10, 47 and 100%, respectively. ROC curves selected the score >6.5 as the best threshold to differentiate between malignant and benign nodules (P < 0.001). The TRS > 6.5 had a better performance than the single parameters evaluated separately, with an accuracy of 77 and 82% upon inclusion of noninvasive follicular thyroid neoplasm with papillary-like nuclear features among malignant or benign cases, respectively. In conclusion, for the first time, we generated a score combining a cost-effective molecular assay with already validated tools, harboring different specificities and sensitivities, for the differential diagnosis of indeterminate nodules. The combination of different parameters reduced the number of false negatives inherent to each classification system. The TRS > 6.5 was highly suggestive for malignancy and retained a high accuracy in the identification of patients to be submitted to surgery.


A Subdivision-Based Representation for Vector Image Editing.
Vector graphics has been employed in a wide variety of applications due to its scalability and editability. Editability is a high priority for artists and designers who wish to produce vector-based graphical content with user interaction. In this paper, we introduce a new vector image representation based on piecewise smooth subdivision surfaces, which is a simple, unified and flexible framework that supports a variety of operations, including shape editing, color editing, image stylization, and vector image processing. These operations effectively create novel vector graphics by reusing and altering existing image vectorization results. Because image vectorization yields an abstraction of the original raster image, controlling the level of detail of this abstraction is highly desirable. To this end, we design a feature-oriented vector image pyramid that offers multiple levels of abstraction simultaneously. Our new vector image representation can be rasterized efficiently using GPU-accelerated subdivision. Experiments indicate that our vector image representation achieves high visual quality and better supports editing operations than existing representations.


Supporting mental health, wellbeing and study skills in Higher Education: an online intervention system.
Dealing with psychological and study skill difficulties can present a challenge for both Higher Education (HE) students, who suffer from them, but also for HE Institutions and their support services. Alternative means of support, such as online interventions, have been identified as cost-effective and efficient ways to provide inclusive support to HE students, removing many of the barriers to help-seeking as well as promoting mental health and wellbeing. The current case study initially outlines the rigorous approach in the development of one such online intervention system, MePlusMe. It further highlights key features that constitute innovative delivery of evidence-based psychological and educational practice in the areas of mental health, promotion of wellbeing, support of mood and everyday functioning, and study-skills enhancement. This case study aims to present the innovative features of MePlusMe in relation to current needs and evidence-basis. Finally, it presents future directions in the evaluation, assessment, and evidence of the fitness-for-purpose process.


Onset of Elasto-capillary Bundling of Micropillar Arrays: A Direct Visualization.
When a liquid drop gets into contact with a soft array of microstructures, capillary forces at the three-phase contact line can lead to critical deformations. Microstructures may collapse and form bundles or even patterns. So far, viewing the kinetics of bundling at the menisci scale has remained elusive. Here, we use laser scanning confocal microscopy to directly image the menisci between micropillars. We image structural changes in polydimethylsiloxane micropillar arrays during the Cassie-to-Wenzel transitions of a water drop evaporating on top of the array. We demonstrate how the regular pillar array undergoes a spontaneous symmetry breaking as the first step to the formation of pillar bundles. A comparison of the Cassie-to-Wenzel transition in air and FC40 indicates that the local contact angle determines the outcome of the bundling process. Based on these observations, we develop a simple model using the local contact angle, stiffness of the pillars, and interfacial tension of the liquid to predict the onset of the symmetry breaking.


[The birth of neuro-psychomotor therapy of developmental age].
The article reconstruct the development of NPMTDA (Neuro-Psychomotor Therapy of Developmental Age) at the University 'La Sapienza' of Rome, where Giovanni Bollea created a team of childhood and adolescence Neuropsychiatry. In the Fifties three physiotherapists coming form the U.S.A., expert in physical, occupational and speech therapy, were the first trainers of roman therapists. Their assessment scheme was adapted to the needs of neuropsychiatric disorders and gradually the above mentioned practices flowed into both evaluation and therapeutic method, thanks also to the competence of other NPCA specialists. A rehabilitation school officially dedicated to children started only in 1973, though it had existed informally since the beginning. It progressively increased in academic dignity finally becoming, in 2000, a graduate school. The professional progress of therapists was favoured by medical advances, National Health Service institution (in the 70's), territorial spreading of rehabilitation centres, academic evolution of NPMTDA teaching.


Mesoporous silica-protected silver nanoparticle disinfectant with controlled Ag+ ion release, efficient magnetic separation, and effective antibacterial activity.
Ag is the most effective metal disinfectant against pathogenic microorganisms and thus, various approaches have been exploited to enhance the dispersity and control the release of Ag+ ions from Ag nanoparticles. In this study, a superparamagnetic Fe3O4@SiO2@Ag@porous SiO2 disinfectant with a double-layer core-shell structure was developed. Its superparamagnetic Fe3O4 nanosphere core ensured its good dispersity in water and allowed its easy magnetic separation after treatment. Its dense SiO2 inner shell protected the Fe3O4 nanosphere core and allowed a good loading of Ag nanoparticles. Its mesoporous SiO2 outer layer effectively protected the Ag nanoparticles from detachment, and its mesoporous channels resulted in lower silver oxidation and dissolution for the controlled release of Ag+ ions. Thus, a highly efficient, silver-based disinfectant was developed, as demonstrated by its effective disinfection of Escherichia coli bacteria with good recycle performance, while the silver concentration in the treated water met the MCL of silver for drinking water.


Diaphragm muscle activity across respiratory motor behaviors in awake and lightly anesthetized rats.
Respiratory muscles such as the diaphragm are active across a range of behaviors including ventilation and higher-force behaviors necessary for maintenance of airway patency, and minimal information is available regarding anesthetic effects on the capacity of respiratory muscles to generate higher forces. The purpose of the present study was to determine whether diaphragm EMG activity during lower-force behaviors, such as eupnea and hypoxia-hypercapnia, is differentially affected compared with higher-force behaviors, such as a sigh, in lightly anesthetized animals. In adult male rats, chronically implanted diaphragm EMG electrodes were used to measure the effects of low-dose ketamine (30 mg/kg) and xylazine (3 mg/kg) on root mean square (RMS) EMG amplitude across a range of motor behaviors. A mixed linear model was used to evaluate the effects of ketamine-xylazine anesthesia on peak RMS EMG and ventilatory parameters, with condition (awake vs. anesthetized), behavior (eupnea, hypoxia-hypercapnia, sigh), side (left or right hemidiaphragm), and their interactions as fixed effects and animal as a random effect. Compared with the awake recordings, there was an overall reduction of peak diaphragm RMS EMG across behaviors during anesthesia, but this reduction was more pronounced during spontaneous sighs (which require ~60% of maximal diaphragm force). Respiratory rates and duty cycle during eupnea and hypoxia-hypercapnia were higher in awake compared with anesthetized conditions. These results highlight the importance of identifying anesthetic effects on a range of respiratory motor behaviors, including sighs necessary for maintaining airway patency. NEW & NOTEWORTHY Respiratory muscles accomplish a range of motor behaviors, with forces generated for ventilatory behaviors comprising only a small fraction of their maximal force generating capacity. Induction of anesthesia exerts more robust effects on the higher-force diaphragm motor behaviors such as sighs compared with eupnea. This novel information on effects of low, sedative doses of a commonly used anesthetic combination (ketamine-xylazine) highlights the importance of identifying anesthetic effects on a range of respiratory motor behaviors.


Multifocal oligodendroglioma in three dogs.
This report describes the clinical, histopathologic, and imaging findings of multifocal oligodendrogliomas from three canine patients. Clinical history varied but included seizure activity and behavior changes. Neurologic examination abnormalities included ataxia, proprioceptive deficits, cranial nerve deficits, and changes in mentation. MRI in one patient revealed multifocal brain lesions; however, the MRI was normal in another one of the patients. Histopathologic evaluation identified multifocal neoplastic infiltrates in all three patients involving the cerebral cortex, brainstem, and spinal cord, with leptomeningeal extension in two of the three patients. All three patients were euthanized due to progression of their neurologic condition and/or complications due to aspiration pneumonia. Oligodendrogliomas should be considered a differential diagnosis for patients with multifocal brain disease.


[Muramyldipeptide - based compounds in current medicine: focus on glucosaminylmuramyl dipeptide].
The role of immune mechanisms in the pathogenesis of almost all human diseases shown in recent decades, increase in antibiotic resistance and secondary immunodeficiency, aging of the population and widespread use of immunosuppressive drugs and procedures suggest a wider use of immunomodulators in current clinical practice, but the use of most of them limits the lack of knowledge. The most promising compounds for the development as immunomodulating agents and adjuvants for a wide range of vaccines are low molecular weight fragments of peptidoglycan - muramylpeptides. The article describes the mechanisms of action of muramylpeptides, their biological effects and properties of medicines developed on their basis. Special emphasis is placed to glucosaminylmuramyl dipeptide registered in the Russian Federation under the trade name Likopid, which is currently the best - studied drug in its group. The results of Likopid studies when used as a prophylactic and therapeutic agent for infections of various localization in adults and children, for oncological diseases and complications of chemotherapy and radiation therapy, psoriasis, atopic and other diseases are presented. It is emphasized that in diseases associated with human papillomavirus and plaque psoriasis, according to current criteria of evidence - based medicine, Likopid should be classified as drug with level A efficacy (high efficiency in 80-100% of patients). High safety of Likopid in adults and children, including newborns, is noted.


Metachronous double spinal dural arteriovenous fistulas. Case report and review of the literature.
Spinal dural arteriovenous fistulas (DAVFs), the most common of spinal vascular malformations, are AVFs in the dura mater of the nerve root and/or adjacent spinal dura. These fistulas are most often solitary and are fed by a single radicular artery that primarily supplies the dura mater. Multiple spinal DAVFs are rarely reported in the literature. Those that have been documented have been synchronous in their presentation in that they were found during the same examination or were present at the initial examination but missed and only recognized at the second examination. The authors report the case of a patient with two spinal DAVFs occurring at different spinal levels at different points in time (metachronous).


Feedback power control strategies in wireless sensor networks with joint channel decoding.
In this paper, we derive feedback power control strategies for block-faded multiple access schemes with correlated sources and joint channel decoding (JCD). In particular, upon the derivation of the feasible signal-to-noise ratio (SNR) region for the considered multiple access schemes, i.e., the multidimensional SNR region where error-free communications are, in principle, possible, two feedback power control strategies are proposed: (i) a classical feedback power control strategy, which aims at equalizing all link SNRs at the access point (AP), and (ii) an innovative optimized feedback power control strategy, which tries to make the network operational point fall in the feasible SNR region at the lowest overall transmit energy consumption. These strategies will be referred to as "balanced SNR" and "unbalanced SNR," respectively. While they require, in principle, an unlimited power control range at the sources, we also propose practical versions with a limited power control range. We preliminary consider a scenario with orthogonal links and ideal feedback. Then, we analyze the robustness of the proposed power control strategies to possible non-idealities, in terms of residual multiple access interference and noisy feedback channels. Finally, we successfully apply the proposed feedback power control strategies to a limiting case of the class of considered multiple access schemes, namely a central estimating officer (CEO) scenario, where the sensors observe noisy versions of a common binary information sequence and the AP's goal is to estimate this sequence by properly fusing the soft-output information output by the JCD algorithm.


The Mechanical Properties of Rat Tail Tendon.
The load-strain and stress-relaxation behavior of wet rat tail tendon has been examined with respect to the parameters strain, rate of straining, and temperature. It is found that this mechanical behavior is reproducible after resting the tendon for a few minutes after each extension so long as the strain does not exceed about 4 per cent. If this strain is exceeded, the tendon becomes progressively easier to extend but its length still returns to the original value after each extension. Extensions of over 35 per cent can be reached in this way. Temperature has no effect upon the mechanical behavior over the range 0-37 degrees C. Just above this temperature, important changes take place in the mechanical properties of the tendon which may have biological significance. The application of the techniques used here to studies of connective tissue disorders is suggested. Some of the mechanical properties of tendon have been interpreted with a simple model.


The perforated tympanic membrane.
Tympanic membrane perforations typically result from trauma or acute otitis media. Most perforations do not cause more than a mild conductive hearing loss, aural fullness and mild tinnitus. Blood, purulent secretions and other debris should be carefully suctioned out of the canal and the perforation size and location described. Irrigation and pneumatic otoscopy should be avoided. A history of vertigo, nausea and vomiting and an audiogram showing a conductive hearing loss of more than 30 dB suggest disruption of the ossicular chain. Profound sensorineural loss may signify inner ear nerve damage. Mastoid radiographs and computed tomographic scans may be useful in cases of significant trauma and infection. Most small perforations resolve spontaneously. The affected ear should be kept dry. Oral and topical antibiotics may be prescribed for perforations related to acute otitis media. Otolaryngologic referral may be necessary to evaluate traumatic perforations associated with vertigo or significant hearing loss, perforations from chronic otitis media or perforations from acute otitis media that do not heal within one month.


Emergence of valley selectivity in monolayer tin(ii) sulphide.
The emergence of valley selectivity in tin(ii) sulphide is explained with the use of density functional theory and the momentum operator matrix elements for the optical transitions. After application of electric stress, the polarization efficiency was found to decrease in the zigzag direction. Wannier functions are further used to derive an effective Tight Binding (TB) model. The velocity matrix elements of the Wannier functions reveal further details about how the p orbitals of Sn and S contribute to optical transitions. Using the TB model in the Wannier basis in a nanoribbon configuration, the bandgap shows an overall decrease as the width of the nanoribbon increases for both zigzag and armchair directions of the structure up to ≈42 Å further presenting opportunities for Optoelectronic applications.


State dependence and temporal evolution of resistance in projected phase change memory.
Phase change memory (PCM) is being actively explored for in-memory computing and neuromorphic systems. The ability of a PCM device to store a continuum of resistance values can be exploited to realize arithmetic operations such as matrix-vector multiplications or to realize the synaptic efficacy in neural networks. However, the resistance variations arising from structural relaxation, 1/f noise, and changes in ambient temperature pose a key challenge. The recently proposed projected PCM concept helps to mitigate these resistance variations by decoupling the physical mechanism of resistance storage from the information-retrieval process. Even though the device concept has been proven successfully, a comprehensive understanding of the device behavior is still lacking. Here, we develop a device model that captures two key attributes, namely, resistance drift and the state dependence of resistance. The former refers to the temporal evolution of resistance, while the latter refers to the dependence of the device resistance on the phase configuration of the phase change material. The study provides significant insights into the role of interfacial resistance in these devices. The model is experimentally validated on projected PCM devices based on antimony and a metal nitride fabricated in a lateral device geometry and is also used to provide guidelines for material selection and device engineering.


A generalized solid-state nudged elastic band method.
A generalized solid-state nudged elastic band (G-SSNEB) method is presented for determining reaction pathways of solid-solid transformations involving both atomic and unit-cell degrees of freedom. We combine atomic and cell degrees of freedom into a unified description of the crystal structure so that calculated reaction paths are insensitive to the choice of periodic cell. For the rock-salt to wurtzite transition in CdSe, we demonstrate that the method is robust for mechanisms dominated either by atomic motion or by unit-cell deformation; notably, the lowest-energy transition mechanism found by our G-SSNEB changes with cell size from a concerted transformation of the cell coordinates in small cells to a nucleation event in large cells. The method is efficient and can be applied to systems in which the force and stress tensor are calculated using density functional theory.


Partnering between monomers of cyclooxygenase-2 homodimers.
Prostaglandin endoperoxide H synthases (PGHSs) 1 and 2 convert arachidonic acid to prostaglandin H2 in the committed step of prostanoid biosynthesis. These enzymes are pharmacological targets of nonsteroidal antiinflammatory drugs and cyclooxygenase (COX) 2 inhibitors. Although PGHSs function as homodimers and each monomer has its own COX and peroxidase active sites, the question of whether there is cross-talk between monomers has remained unresolved. Here we describe two heterodimers in which a native subunit of human PGHS-2 has been coupled to a subunit having a defect within the COX active site at some distance from the dimer interface. Native/G533A PGHS-2, a heterodimer with a COX-inactive subunit, had the same specific COX activity as the native homodimer. Native/R120Q PGHS-2, a heterodimer in which both subunits can oxygenate arachidonic acid but in which the R120Q subunit cannot bind the COX inhibitor flurbiprofen, was inhibited by flurbiprofen to about the same extent as native PGHS-2. These results imply that native PGHS-2 exhibits half-of-sites reactivity. Isothermal titration calorimetry established that only one monomer of the native PGHS-2 homodimer binds flurbiprofen tightly. In short, binding of ligand to the COX site of one monomer alters its companion monomer so that it is unable to bind substrate or inhibitor. We conclude that PGHS monomers comprising a dimer, although identical in the resting enzyme, differ from one another during catalysis. The nonfunctioning subunit may provide structural support enabling its partner monomer to catalyze the COX reaction. This subunit complementarity may prove to be characteristic of other dimeric enzymes having tightly associated monomers.


Atorvastatin 10 mg plus ezetimibe 10mg compared with atorvastatin 20 mg: impact on the lipid profile in Japanese patients with abnormal glucose tolerance and coronary artery disease.
Oxidized low-density lipoprotein (LDL) cholesterol is a sensitive lipid marker for predicting atherosclerosis. Ezetimibe and statins are reported to decrease both LDL cholesterol and oxidized LDL cholesterol. This prospective randomized open-label crossover study compared combination therapy with atorvastatin plus ezetimibe versus high-dose atorvastatin monotherapy. Changes in serum lipids, including malondialdehyde-modified LDL (MDA-LDL) as a representative form of oxidized LDL cholesterol, and glucose metabolism were assessed. The subjects were 39 Japanese patients with coronary artery disease and type 2 diabetes or impaired glucose tolerance who were taking 10 mg/day of atorvastatin (30 men and 9 women with a mean age of 67.8 years). They were randomized to a group that first received add-on ezetimibe (10 mg/day) or a group that first received atorvastatin monotherapy at a higher dose of 20 mg/day. Both treatments were given for 12 weeks each in a crossover fashion. Add-on ezetimibe significantly decreased MDA-LDL (109.0 ± 31.9 mg/dl to 87.7 ± 29.4 mg/dl, p=0.0009), while up-titration of atorvastatin did not. The decrease with add-on ezetimibe was significantly greater than with up-titration of atorvastatin (p=0.0006). Total cholesterol and LDL cholesterol were significantly decreased by both treatments, but the percent reduction with add-on ezetimibe was significantly greater (p<0.05). High-density lipoprotein cholesterol was significantly increased by both treatments and there was no significant difference between them. The apolipoprotein B/apolipoprotein A-I ratio and remnant-like particle cholesterol were only significantly decreased by add-on ezetimibe. Both treatments caused similar elevation of hemoglobin A(1c). In Japanese patients with type 2 diabetes or impaired glucose tolerance and coronary artery disease, adding ezetimibe (10 mg/day) to atorvastatin (10 mg/day) significantly improved the lipid profile compared with atorvastatin monotherapy at 20 mg/day.


Evaluation of the two bundles of the anterior cruciate ligament with 1.5 tesla magnetic resonance imaging.
Studies on the anatomy of the anterior cruciate ligament (ACL) have shown that the normal ACL consists of two functional bundles named as anteromedial (AM) and posterolateral (PL) bundles. In this study, we evaluated the AM and PL bundles of the ACL using 1.5 tesla magnetic resonance imaging (MRI), which is routinely used in clinical practice. The study included 150 patients (96 females, 54 males; mean age 33.4+/-11.6 years; range 18 to 59 years) who did not have any signs of ACL insufficiency and whose knees were examined by MRI for other reasons. Standard magnetic resonance images (77 right, 73 left) were evaluated independently by an orthopedist and a radiologist in terms of distinguishable ACL bundles. The angle between the ACL (and each bundle) and the tibial plateau was measured on sagittal and coronal sections. Arthroscopic surgery was performed in 64 patients (42.7%) for primary diagnoses and arthroscopic and MRI findings were compared. Magnetic resonance imaging showed an intact ACL in all the patients. The ACL was assessed as a single bundle in the axial, coronal, and sagittal planes in 93 patients (62%). A double-bundle appearance was noted in 57 patients (38%), involving all three planes in 14 patients (9.3%), axial and coronal planes in 41 patients (27.3%), coronal and sagittal planes in one patient (0.7%), and only coronal plane in one patient (0.7%). On MRI sections showing a single bundle ACL, the mean angle between the ACL and the tibial plateau was found as 55.3 degrees in the sagittal plane, and 70.3 degrees in the coronal plane. On sections with a double-bundle appearance, the mean angles between the AM bundle and the tibial plateau were 70.1 degrees and 55.1 degrees in the coronal and sagittal planes, respectively. The corresponding angles for the PL bundle were 81 degrees and 53.5 degrees . The incidence of double bundle ACL appearance in coronal, sagittal, and axial MRI sections was not influenced by sex and side (p>0.05). The number of bundles identified in each plane did not show a significant difference between the two observers (p>0.05). During arthroscopic surgery, both bundles were identified with normal integrity and function of the ACL in all the patients. Of these, MRI could depict a double-bundle appearance in one or more planes in only 42.2% of the patients. Even though standard 1.5 tesla MRI, routinely used in clinical practice, has a very high success rate in demonstrating the ACL, it can visualize the two-bundle structure only in about one-third of the patients.


The impact of resistance training on brain-derived neurotrophic factor and depression among older adults aged 60 years or older: A systematic review and meta-analysis of randomized controlled trials.
This systematic review and meta-analysis aimed to investigate the impact of resistance training on brain-derived neurotrophic factor (BDNF) and depression among older adults aged 60 years or older. Four electronic databases were systematically searched. A total of 11 randomized controlled trials, with a pooled sample of 868 participants, met our inclusion criteria. Meta-analysis demonstrated that resistance training significantly improved circulating BDNF levels (mean difference; MD: 0.73 ng/ml; 95% CI [0.04, 1.42]; p = 0.04). Additionally, resistance training was associated with significant improvements in depression (standardized mean difference; SMD: -0.38; 95% CI [- 0.62, -0.14]; p = 0.002). These findings suggest that resistance training may be an effective intervention for improving BDNF levels and reducing depression symptoms in older adults. Further research is needed to confirm these findings and to investigate the underlying mechanisms.


Magnitude of preoperative cervical lordotic compensation and C2-T3 angle are correlated to increased risk of postoperative sagittal spinal pelvic malalignment in adult thoracolumbar deformity patients at 2-year follow-up.
Cervical deformity (CD) is prevalent among patients with adult spinal deformity (ASD). The effect of baseline cervical alignment on achieving optimal thoracolumbar alignment in ASD surgery is unclear. This study assesses the relationship between preoperative (preop) cervical spinal parameters and global alignment after thoracolumbar ASD surgery at 2-year follow-up. This study is a retrospective review of a multicenter, prospective database. Surgical ASD patients with 2-year follow-up and cervical X-rays were included. The outcome measures were radiographic parameters and self-reported health-related quality-of-life measures (Short-Form 36 [SF-36], Oswestry Disability Index [ODI], and Scoliosis Research Society 22 [SRS-22]). Surgical ASD patients of 18 years and older with scoliosis greater than or equal to 20° and one of the following radiographic parameters were included: sagittal vertical axis (SVA) greater than or equal to 5 cm, pelvic tilt (PT) greater than or equal to 25°, or thoracic kyphosis (TK) greater than 60°. The SRS-Schwab sagittal modifiers (PT, global alignment, and pelvic incidence and lumbar lordosis [PI-LL]) were assessed at 2-year postoperatively as either normal ("0") or abnormal ("+" or "++"). Patients were classified in the aligned group (AG) or malaligned group (MG) at 2-year follow-up if all three sagittal modifiers were normal or abnormal, respectively. Patients were assessed for CD based on the following criteria: C2-C7 SVA greater than 4 cm, C2-C7 SVA less than 4 cm, cervical kyphosis (CL greater than 0), cervical lordosis (CL less than 0), any deformity (C2-C7 SVA greater than 4 cm or CL greater than 0), and both CD (C2-C7 SVA greater than 4 cm and CL greater than 0). Univariate testing was performed using t or chi-square test, looking at the following preop parameters: CD, C2-C7 SVA, C2-T3 SVA, CL, T1 slope (T1S), T1S-CL, C2-T3 angle, LL, TK, PT, C7-S1 SVA, and PI-LL. One hundred four patients met the initial inclusion criteria with 70 in the AG and 34 in MG. Preoperative, patients in the MG had a higher CL (11.7 vs. 4.9, p=.03), higher C2-T3 angle (13.59 vs 4.9 p=.01), higher PT (p<.0001), higher SVA (p<.0001), and higher PI-LL (p<.0001) compared with the AG. Interestingly, the prevalence of CD at baseline was similar for both groups. There was no statistically significant difference among groups in the amount of improvement more than 2 years on the ODI or the Physical Component Summary of SF-36. Patients with sagittal spinal malalignment associated with significant cervical compensatory lordosis are at increased risk of realignment failure at 2-year follow-up. Assessment of the degree of cervical compensation may be helpful in preop evaluation to assist in realignment outcome prediction.


Bridging 3D Slicer and ROS2 for Image-Guided Robotic Interventions.
Developing image-guided robotic systems requires access to flexible, open-source software. For image guidance, the open-source medical imaging platform 3D Slicer is one of the most adopted tools that can be used for research and prototyping. Similarly, for robotics, the open-source middleware suite robot operating system (ROS) is the standard development framework. In the past, there have been several "ad hoc" attempts made to bridge both tools; however, they are all reliant on middleware and custom interfaces. Additionally, none of these attempts have been successful in bridging access to the full suite of tools provided by ROS or 3D Slicer. Therefore, in this paper, we present the SlicerROS2 module, which was designed for the direct use of ROS2 packages and libraries within 3D Slicer. The module was developed to enable real-time visualization of robots, accommodate different robot configurations, and facilitate data transfer in both directions (between ROS and Slicer). We demonstrate the system on multiple robots with different configurations, evaluate the system performance and discuss an image-guided robotic intervention that can be prototyped with this module. This module can serve as a starting point for clinical system development that reduces the need for custom interfaces and time-intensive platform setup.


Duplex ultrasonography, digital subtraction angiography, and conventional angiography in assessing carotid atherosclerosis.
We prospectively compared duplex ultrasonography with digital subtraction angiography (DSA) and conventional angiography in the evaluation of carotid atherosclerosis by studying 494 arteries. Arteries were graded independently as normal, mildly stenotic, moderately stenotic, severely stenotic, or occluded. There was absolute agreement between duplex ultrasonography and DSA as to disease severity in 296 (68%) of 434 vessels, good correlation in an additional 119 vessels (28%), and poor correlation in 19 vessels (4%). When duplex studies were compared with conventional angiograms, the correlations were similar: absolute for 60 (71%) of 85 vessels, good for 17 (20%) of 85 vessels, and poor for eight (9%) of 85 vessels. With conventional angiography as the standard, duplex studies tended to overestimate the severity of disease in 64% of vessels and underestimate it in 36%, v 28% and 72%, respectively, for DSA. We concluded that duplex ultrasonography is as accurate as DSA in assessing carotid bifurcation atherosclerosis. The tendency of DSA to underestimate disease severity may lead to diagnostic errors in patients with nonobstructive plaques.


Increased Slc12a1 expression in β-cells and improved glucose disposal in Slc12a2 heterozygous mice.
The products of the Slc12a1 and Slc12a2 genes, commonly known as Na(+)-dependent K(+)2Cl(-) co-transporters NKCC2 and NKCC1, respectively, are the targets for the diuretic bumetanide. NKCCs are implicated in the regulation of intracellular chloride concentration ([Cl(-)]i) in pancreatic β-cells, and as such, they may play a role in glucose-stimulated plasma membrane depolarization and insulin secretion. Unexpectedly, permanent elimination of NKCC1 does not preclude insulin secretion, an event potentially linked to the homeostatic regulation of additional Cl(-) transporters expressed in β-cells. In this report we provide evidence for such a mechanism. Mice lacking a single allele of Slc12a2 exhibit lower fasting glycemia, increased acute insulin response (AIR) and lower blood glucose levels 15-30 min after a glucose load when compared to mice harboring both alleles of the gene. Furthermore, heterozygous expression or complete absence of Slc12a2 associates with increased NKCC2 protein expression in rodent pancreatic β-cells. This has been confirmed by using chronic pharmacological down-regulation of NKCC1 with bumetanide in the mouse MIN6 β-cell line or permanent molecular silencing of NKCC1 in COS7 cells, which results in increased NKCC2 expression. Furthermore, MIN6 cells chronically pretreated with bumetanide exhibit increased initial rates of Cl(-) uptake while preserving glucose-stimulated insulin secretion. Together, our results suggest that NKCCs are involved in insulin secretion and that a single Slc12a2 allele may protect β-cells from failure due to increased homeostatic expression of Slc12a1.


ADAM17 Inhibitors Attenuate Corneal Epithelial Detachment Induced by Mustard Exposure.
Sulfur mustard, nitrogen mustard (NM), and 2-chloroethyl ethyl sulfide all cause corneal injury with epithelial-stromal separation, differing only by degree. Injury can resolve in a few weeks or develop into chronic corneal problems. These vesicants induce microbullae at the epithelial-stromal junction, which is partially caused by cleavage of transmembranous hemidesmosomal collagen XVII, a component anchoring the epithelium to the stroma. ADAM17 is an enzyme involved in wound healing and is able to cleave collagen XVII. The activity of ADAM17 was inhibited in vesicant-exposed corneas by four different hydroxamates, to evaluate their therapeutic potential when applied 2 hours after exposure, thereby allowing ADAM17 to perform its early steps in wound healing. Rabbit corneal organ cultures exposed to NM for 2 hours were washed, then incubated at 37°C for 22 hours, with or without one of the four hydroxamates (dose range, 0.3-100 nmol in 20 μL, applied four times). Corneas were analyzed by light and immunofluorescence microscopy, and ADAM17 activity assays. Nitrogen mustard-induced corneal injury showed significant activation of ADAM17 levels accompanying epithelial-stromal detachment. Corneas treated with hydroxamates starting 2 hours post exposure showed a dose-dependent ADAM17 activity inhibition up to concentrations of 3 nmol. Of the four hydroxamates, NDH4417 (N-octyl-N-hydroxy-2-[4-hydroxy-3-methoxyphenyl] acetamide) was most effective for inhibiting ADAM17 and retaining epithelial-stromal attachment. Mustard exposure leads to corneal epithelial sloughing caused, in part, by the activation of ADAM17 at the epithelial-stromal junction. Select hydroxamate compounds applied 2 hours after NM exposure mitigated epithelial-stromal separation.


A novel ARID DNA-binding protein interacts with SymRK and is expressed during early nodule development in Lotus japonicus.
During the establishment of symbiosis in legume roots, the rhizobial Nod factor signal is perceived by the host cells via receptor-like kinases, including SymRK. The NODULE INCEPTION (NIN) gene in Lotus japonicus is required for rhizobial entry into root cells and for nodule organogenesis. We describe here a novel DNA-binding protein from L. japonicus, referred to as SIP1, because it was identified as a SymRK-interacting protein. SIP1 contains a conserved AT-rich interaction domain (ARID) and represents a unique member of the ARID-containing proteins in plants. The C terminus of SIP1 was found to be responsible for its interaction with the kinase domain of SymRK and for homodimerization in the absence of DNA. SIP1 specifically binds to the promoter of LjNIN but not to that of LjCBP1 (a calcium-binding protein gene), both of which are known to be inducible by Nod factors. SIP1 recognizes two of the three AT-rich domains present in the NIN gene promoter. Deletion of one of the AT-rich domains at the NIN promoter diminishes the binding of SIP1 to the NIN promoter. The protein is localized to the nuclei when expressed as a red fluorescence fusion protein in the onion (Allium cepa) epidermal cells. The SIP1 gene is expressed constitutively in the uninfected roots, and its expression levels are elevated after infection by Mesorhizobium loti. It is proposed that SIP1 may be required for the expression of NIN and involved in the initial communications between the rhizobia and the host root cells.


Instrumental variable estimation of early treatment effect in randomized screening trials.
The primary analysis of randomized screening trials for cancer typically adheres to the intention-to-screen principle, measuring cancer-specific mortality reductions between screening and control arms. These mortality reductions result from a combination of the screening regimen, screening technology and the effect of the early, screening-induced, treatment. This motivates addressing these different aspects separately. Here we are interested in the causal effect of early versus delayed treatments on cancer mortality among the screening-detectable subgroup, which under certain assumptions is estimable from conventional randomized screening trial using instrumental variable type methods. To define the causal effect of interest, we formulate a simplified structural multi-state model for screening trials, based on a hypothetical intervention trial where screening detected individuals would be randomized into early versus delayed treatments. The cancer-specific mortality reductions after screening detection are quantified by a cause-specific hazard ratio. For this, we propose two estimators, based on an estimating equation and a likelihood expression. The methods extend existing instrumental variable methods for time-to-event and competing risks outcomes to time-dependent intermediate variables. Using the multi-state model as the basis of a data generating mechanism, we investigate the performance of the new estimators through simulation studies. In addition, we illustrate the proposed method in the context of CT screening for lung cancer using the US National Lung Screening Trial data.


Enhanced ROCK1 dependent contractility in fibroblast from chronic obstructive pulmonary disease patients.
During wound healing processes fibroblasts account for wound closure by adopting a contractile phenotype. One disease manifestation of COPD is emphysema which is characterized by destruction of alveolar walls and our hypothesis is that fibroblasts in the COPD lungs differentiate into a more contractile phenotype as a response to the deteriorating environment. Bronchial (central) and parenchymal (distal) fibroblasts were isolated from lung explants from COPD patients (n = 9) (GOLD stage IV) and from biopsies from control subjects and from donor lungs (n = 12). Tissue-derived fibroblasts were assessed for expression of proteins involved in fibroblast contraction by western blotting whereas contraction capacity was measured in three-dimensional collagen gels. The basal expression of rho-associated coiled-coil protein kinase 1 (ROCK1) was increased in both centrally and distally derived fibroblasts from COPD patients compared to fibroblasts from control subjects (p < 0.001) and (p < 0.01), respectively. Distally derived fibroblasts from COPD patients had increased contractile capacity compared to control fibroblasts (p < 0.01). The contraction was dependent on ROCK1 activity as the ROCK inhibitor Y27632 dose-dependently blocked contraction in fibroblasts from COPD patients. ROCK1-positive fibroblasts were also identified by immunohistochemistry in the alveolar parenchyma in lung tissue sections from COPD patients. Distally derived fibroblasts from COPD patients have an enhanced contractile phenotype that is dependent on ROCK1 activity. This feature may be of importance for the elastic dynamics of small airways and the parenchyma in late stages of COPD.


How Do You Feel the Rhythm: Dynamic Motor-Auditory Interactions Are Involved in the Imagination of Hierarchical Timing.
Predicting and organizing patterns of events is important for humans to survive in a dynamically changing world. The motor system has been proposed to be actively, and necessarily, engaged in not only the production but the perception of rhythm by organizing hierarchical timing that influences auditory responses. It is not yet well understood how the motor system interacts with the auditory system to perceive and maintain hierarchical structure in time. This study investigated the dynamic interaction between auditory and motor functional sources during the perception and imagination of musical meters. We pursued this using a novel method combining high-density EEG, EMG, and motion capture with independent component analysis to separate motor and auditory activity during meter imagery while robustly controlling against covert movement. We demonstrated that endogenous brain activity in both auditory and motor functional sources reflects the imagination of binary and ternary meters in the absence of corresponding acoustic cues or overt movement at the meter rate. We found clear evidence for hypothesized motor-to-auditory information flow at the beat rate in all conditions, suggesting a role for top-down influence of the motor system on auditory processing of beat-based rhythms, and reflecting an auditory-motor system with tight reciprocal informational coupling. These findings align with and further extend a set of motor hypotheses from beat perception to hierarchical meter imagination, adding supporting evidence to active engagement of the motor system in auditory processing, which may more broadly speak to the neural mechanisms of temporal processing in other human cognitive functions.SIGNIFICANCE STATEMENT Humans live in a world full of hierarchically structured temporal information, the accurate perception of which is essential for understanding speech and music. Music provides a window into the brain mechanisms of time perception, enabling us to examine how the brain groups musical beats into, for example a march or waltz. Using a novel paradigm combining measurement of electrical brain activity with data-driven analysis, this study directly investigates motor-auditory connectivity during meter imagination. Findings highlight the importance of the motor system in the active imagination of meter. This study sheds new light on a fundamental form of perception by demonstrating how auditory-motor interaction may support hierarchical timing processing, which may have clinical implications for speech and motor rehabilitation.


Effects of free fatty acids on meibomian lipid films.
The purpose of this study was to evaluate the impact of free fatty acids (FFA), namely oleic (OA) and linoleic (LA) ones, on meibomian lipid films (MLF) using a Langmuir trough (LT) and a Brewster angle microscope (BAM). Human meibum was collected from healthy volunteers. A Tris-buffered saline (TBS, pH 7.4) was used as the control aqueous subphase for LT experiments. Then, varying amounts of OA and LA were dissolved in TBS to make FFA-containing subphases. Predetermined amounts of meibum were loaded onto the surface of the (TBS/±FFA) subphases to form MLF. Then, surface pressure-area (π/A) isotherms of MLF were recorded. Standard rheological parameters such as rigidity, elasticity, and hysteresis, were computed. In a separate experiment, OA and LA were pre-mixed with meibum at different weight ratios prior their spreading onto the control TBS subphase, and the (π/A) isotherms of the resulting mixed films of meibum and FFA were studied and analyzed in the same fashion as described above. When studied at the normal corneal temperature of 34 °C with the (TBS/-FFA) subphase, meibum formed stable films. When (TBS/+FFA) subphase was used, both FFA quickly disrupted the MLF, acting in a similar fashion. BAM revealed that the most dramatic changes in the structure of MLF occurred in the range of OA concentrations between 5 and 15 μM. However, this effect was apparent even with 2.5 μM OA. When OA was pre-mixed with meibum, but was absent from the subphase, it caused gradual concentration-dependent changes in the (π/A) isotherms, but the MLF did not disappear from the surface. Thus, tested FFA showed a remarkable ability to disrupt, and/or prevent the formation of, human MLF, which could contribute to the onset of those forms of dry eye disease that are associated with enhanced activity of lipolytic enzymes, such as chronic blepharitis.


Efficacy of combination-chemotherapy with pirarubicin, ifosfamide, and etoposide for soft tissue sarcoma: a single-institution retrospective analysis.
The standard chemotherapy regimens for soft tissue sarcoma are doxorubicin-based. This retrospective study aimed to assess the efficacy and safety of pirarubicin, ifosfamide, and etoposide combination therapy for patients with this disease. Between 2008 and 2017, 25 patients with soft tissue sarcoma were treated with pirarubicin (30 mg/m2, 2 days), ifosfamide (2 g/m2, 5 days), and etoposide (100 mg/m2, 3 days) every 3 weeks. The primary endpoint was overall response, and the secondary endpoint was adverse events of this regimen. Responses to this regimen according to RECIST criteria were partial response (n = 9, 36%), stable disease (n = 9, 36%) and progressive disease (n = 7, 28%). During the treatment phase, frequent grade 3 or worse adverse events were hematological toxicities including white blood cell decreases (96%), febrile neutropenia (68%), anemia (68%), and platelet count decreases (48%). No long-term adverse events were reported during the study period. This regimen was comparable to previously published doxorubicin-based combination chemotherapy in terms of response rate. Although there were no long-lasting adverse events, based on our results, severe hematological toxicity should be considered.


The hidden cost of audit.
Although a considerable sum of money has been made available for the introduction of medical audit, many of the indirect costs have not been considered. We have assessed one of these areas, the medical audit meeting, by calculating the cost to Preston Acute Hospitals (PAH) Trust and extrapolating it to England and Wales. A potential cost of Pounds 678,000 per annum to PAH Trust could be reduced to 14 per cent of this figure if current practices were changed. The hidden cost for England and Wales could be up to Pounds 106 million per annum. The cost of medical audit meetings could be dramatically reduced if current practices were modified. This could be done with little detriment to the audit process.


Induction of thymic stromal lymphopoietin by a steroid alkaloid derivative in mouse keratinocytes.
Thymic stromal lymphopoietin (TSLP) plays critical roles in inducing and exacerbating allergic diseases. Chemical compounds that induce TSLP production can enhance sensitization to antigens and exacerbate allergic inflammation. Hence, identifying such chemicals will be important to prevent an increase in allergic diseases. In the present study, we found, for the first time, that a steroid alkaloid derivative, code no. 02F04, concentration and time dependently induced mRNA expression and production of TSLP in a mouse keratinocyte cell line, PAM212. In particular, the activity of 02F04 was selective to TSLP. As an analogue of the liver X receptor (LXR) endogenous ligand, 02F04 rapidly increased ATP-binding cassette transporter A1 (ABCA1) expression by regulating the nuclear receptor of LXR. However, instead of being inhibited by the LXR antagonist, 02F04-induced TSLP production was delayed and markedly suppressed by inhibitors of phospholipase C (PLC), pan-protein kinase C (PKC), PKCδ, Rho-associated protein kinase (ROCK), extracellular signal-regulated kinase (ERK) 1/2, and IκΒ kinase 2 (IKK2). Treatment with 02F04 caused the formation of F-actin filaments surrounding the nucleus of PAM212 cells, which then disappeared following addition of ROCK inhibitor. 02F04 also induced phosphorylation of ERK1/2 from 2h after treatment, with a maximum at 24h, and increased nuclear factor-κB (NF-κB) promoter activity by 1.3-fold. Taken together, these results indicate that 02F04-induced TSLP production is regulated via distinct signal transduction pathways, including PLC, PKC, ROCK, ERK1/2, and NF-κB but not nuclear receptors. 02F04, with a unique skeletal structure in inducing TSLP production, can represent a potential new tool for investigating the role of TSLP in allergic diseases.


Cutting edge: mast cells critically augment myeloid-derived suppressor cell activity.
Myeloid-derived suppressor cells (MDSCs) are primarily recognized for their immunosuppressive properties in malignant disease. However, their interaction with other innate immune cells and their regulation of immune responses, such as in parasitic infection, necessitate further characterization. We used our previously published mouse model of MDSC accumulation to examine the immunoregulatory role of MDSCs in B16 melanoma metastasis and Nippostrongylus brasiliensis infection. In this study, we demonstrate that the activity of MDSCs is dependent on the immune stimuli and subset induced. Monocytic MDSCs predictably suppressed antitumor immune responses but granulocytic MDSCs surprisingly enhanced the clearance of N. brasiliensis infection. Intriguingly, both results were dependent on MDSC interaction with mast cells (MCs), as demonstrated by adoptive-transfer studies in MC-deficient (Kit(Wsh)(/)(Wsh)) mice. These findings were further supported by ex vivo cocultures of MCs and MDSCs, indicating a synergistic increase in cytokine production. Thus, MCs can enhance both immunosuppressive and immunosupportive functions of MDSCs.


Use of Technology-Based Tools to Support Adolescents and Young Adults With Chronic Disease: Systematic Review and Meta-Analysis.
With the large amount of material that is readily available on the internet, there are endless opportunities for electronic health-literate patients to obtain and learn new information. Although novel, a Web- or mobile-based program can be a powerful way to engage adolescents and young adults (AYAs). The ongoing engagement of AYAs with chronic disease is vital not only to empower them but also to ensure a smooth transition from pediatric to adult health care. This study aimed to evaluate the current evidence on Web- or mobile-based interventions designed for AYAs. This review was registered with PROSPERO: CRD42018096487. A systematic search of MEDLINE Complete, EMBASE, and CINAHL Complete was conducted on April 10, 2019, for studies that examined the perspectives of transition-age patients about technology-based interventions, the process involved in intervention development, or the evaluation of intervention efficacy. For each study, the comprehensiveness of reporting was appraised. The Downs and Black checklist was used for intervention efficacy trials, the Standards for Reporting Qualitative Research checklist was used for qualitative work, and a 16-item tool developed by Tong et al was used for questionnaire research. The search uncovered 29 relevant studies, which included qualitative studies (n=14), intervention efficacy studies (n=7), questionnaire studies (n=4), mixed qualitative and questionnaire studies (n=2), and a mixed qualitative and pilot randomized controlled trial study (n=1). The reporting comprehensiveness score of questionnaires was rated considerably lower (n=6, 13%-57% [2/16-8/14]) than the scores of intervention efficacy trials (n=8, 48%-85% [13/27-23/27]) and qualitative research (n=17, 40%-93% [8.5/21-19.5/21]). AYAs were receptive to obtaining information via a website or mobile app. An intervention was more likely to be perceived as useful by AYAs when there was a concerted effort to involve AYAs and subject matter experts in the process of intervention design, as opposed to relying solely on the AYAs or the experts alone. The preferred medium of intervention delivery varied greatly for AYAs, ranging from static text to audiovisual materials. However, AYAs considered being concise was the most important aspect. Across different conditions, AYAs were interested in receiving information on diverse topics, such as anxiety and stress management, dealing with insurance, and having social relationships. Patients also requested for disease-specific information, such as weather forecasts and pollen levels for patients with asthma and information related to the pretransplant period for organ transplant recipients. Meta-analyses showed no significant group differences across time on quality of life, self-efficacy, and self-management. Owing to the lack of intervention efficacy trials, no conclusion can be drawn if an intervention delivered via a mobile app is better than that delivered via a website. However, through this systematic review, it is confirmed that AYAs were receptive to receiving medical information electronically.


Protective effect of endogenous beta-adrenergic tone on lung fluid balance in acute bacterial pneumonia in mice.
Some investigators have reported that endogenous beta-adrenoceptor tone can provide protection against acute lung injury. Therefore, we tested the effects of beta-adrenoceptor inhibition in mice with acute Escherichia coli pneumonia. Mice were pretreated with propranolol or saline and then intratracheally instilled with live E. coli (10(7) colony-forming units). Hemodynamics, arterial blood gases, plasma catecholamines, extravascular lung water, lung permeability to protein, bacterial counts, and alveolar fluid clearance were measured. Acute E. coli pneumonia was established after 4 h with histological evidence of acute pulmonary inflammation, arterial hypoxemia, a threefold increase in lung vascular permeability, and a 30% increase in extravascular lung water as an increase in plasma catecholamine levels. beta-Adrenoceptor inhibition resulted in a marked increase in extravascular lung water that was explained by both an increase in lung vascular permeability and a reduction in net alveolar fluid clearance. The increase in extravascular lung water with propranolol pretreatment was not explained by an increase in systemic or vascular pressures. The increase in lung vascular permeability was explained in part by anti-inflammatory effects of beta-adrenoceptor stimulation because plasma macrophage inflammatory protein-2 levels were higher in the propranolol pretreatment group compared with controls. The decrease in alveolar fluid clearance with propranolol was explained by a decrease in catecholamine-stimulated fluid clearance. Together, these results indicate that endogenous beta-adrenoceptor tone has a protective effect in limiting accumulation of extravascular lung water in acute severe E. coli pneumonia in mice by two mechanisms: 1) reducing lung vascular injury and 2) upregulating the resolution of alveolar edema.


Cell morphology, markers, spreading, and proliferation on orthopaedic biomaterials. An innovative cellular model for the "in vitro" study.
The aims of tissue engineering are the in vitro reconstruction of functionally active tissues, and the in vivo induction of their appropriate development. The great progresses in the fields of biology and biomaterials represent key events, which allowed the recent improvement of tissue engineering. In the orthopaedic perspective, tissue engineering is focused on the development of innovative materials, whose action consists in recruiting bone progenitor cells and in stimulating their proliferation. In this context, it should remind that these materials should not only allow cells adhesion and proliferation, but also ensure that attached cells maintain the cellular properties of the original tissue. In this study, a new cellular model, suitable for the rapid in vitro determination of the above parameters, is presented. The cell model derives from a human osteosarcoma cell line, Saos-2, which maintained the cytological features of the osteoblast cells. The cell line was genetically modified to express constitutively the enhanced green fluorescent protein. The engineered cell line Saos-eGFP represents a suitable in vitro mode for studying the biocompatibility, the cell adhesion, spreading, and proliferation on biomaterials developed for clinical applications.


SPO11.2 is essential for programmed double-strand break formation during meiosis in bread wheat (Triticum aestivum L.).
Meiotic recombination is initiated by formation of DNA double-strand breaks (DSBs). This involves a protein complex that includes in plants the two similar proteins, SPO11-1 and SPO11-2. We analysed the sequences of SPO11-2 in hexaploid bread wheat (Triticum aestivum), as well as in its diploid and tetraploid progenitors. We investigated its role during meiosis using single, double and triple mutants. The three homoeologous SPO11-2 copies of hexaploid wheat exhibit high nucleotide and amino acid similarities with those of the diploids, tetraploids and Arabidopsis. Interestingly, however, two nucleotides deleted in exon-2 of the A copy lead to a premature stop codon and suggest that it encodes a non-functional protein. Remarkably, the mutation was absent from the diploid A-relative Triticum urartu, but present in the tetraploid Triticum dicoccoides and in different wheat cultivars indicating that the mutation occurred after the first polyploidy event and has since been conserved. We further show that triple mutants with all three copies (A, B, D) inactivated are sterile. Cytological analyses of these mutants show synapsis defects, accompanied by severe reductions in bivalent formation and numbers of DMC1 foci, thus confirming the essential role of TaSPO11-2 in meiotic recombination in wheat. In accordance with its 2-nucleotide deletion in exon-2, double mutants for which only the A copy remained are also sterile. Notwithstanding, some DMC1 foci remain visible in this mutant, suggesting a residual activity of the A copy, albeit not sufficient to restore fertility.


Idiopathic ventricular fibrillation: the ongoing quest for diagnostic refinement.
Prior to the recognition of distinct clinical entities, such as Brugada syndrome, catecholaminergic polymorphic ventricular tachycardia, and long QT syndrome, all sudden cardiac arrest (SCA) survivors with ventricular fibrillation (VF) and apparently structurally normal hearts were labelled as idiopathic ventricular fibrillation (IVF). Over the last three decades, the definition of IVF has changed substantially, mostly as result of the identification of the spectrum of SCA-predisposing genetic heart diseases (GHDs), and the molecular evidence, by post-mortem genetic analysis (aka, the molecular autopsy), of cardiac channelopathies as the pathogenic basis for up to 35% of unexplained cases of sudden cardiac death (SCD) in the young. The evolution of the definition of IVF over time has led to a progressively greater awareness of the need for an extensive diagnostic assessment in unexplained SCA survivors. Nevertheless, GHDs are still underdiagnosed among SCA survivors, due to the underuse of pharmacological challenges (i.e. sodium channel blocker test), misrecognition of electrocardiogram (ECG) abnormalities/patterns (i.e. early repolarization pattern or exercise-induced ventricular bigeminy) or errors in the measurement of ECG parameters (e.g. the heart-rate corrected QT interval). In this review, we discuss the epidemiology, diagnostic approaches, and the controversies related to role of the genetic background in unexplained SCA survivors with a default diagnosis of IVF.


Temperature-dependent molecular motions of cholesterol esters: a carbon-13 nuclear magnetic resonance study.
Carbon-13 NMR spectroscopy at 50.3 MHz has been used to study four long-chain cholesterol esters with a double bond in the omega-9 position: cholesteryl oleate, C18:1, omega-9; cholesteryl linoleate, C18:2, omega-6,9; cholesteryl erucate, C22:1, omega-9; cholesteryl nervonate, C24:1, omega-9. The linoleate and oleate esters exhibit two metastable liquid-crystalline phases (cholesteric and smectic), whereas the longer chain esters form a stable smectic phase but no cholesteric phase [Ginsburg, G. S., & Small, D. M. (1981) Biochim. Biophys. Acta 664, 98-107]. Line widths (nu 1/2), spin--lattice relaxation times (T1), and nuclear Overhauser enhancements (NOE) were measured for all well-resolved resonances from ring and fatty acyl (FA) carbons at different temperatures in the isotropic liquid of each ester. T1 and NOE values of FA resonances were constant between the FA-2 carbon and olefinic region of each acyl chain and increased markedly for carbons near the chain terminus. FA carbon motions are thus restricted and/or highly correlated in the region between the ring and the olefinic carbons, suggesting that strong interactions occur between cholesterol ester molecules in this region of the FA chain. These results also suggest that the FA chains are approximately extended in the isotropic liquid. Steroid ring methine C-6 and C-3 nu 1/2's increased differentially on cooling to the liquid leads to liquid crystal transition temperature (Tm) of each ester, indicative of increasingly anisotropic ring rotations. The rotational anisotropy was quantitated by using a prolate ellipsoid model for the cholesterol ester molecule for which two correlation times (corresponding to rotations about the long and short molecular axes) were calculated from the C-3 and C-6 nu 1/2 values. The C-3/C-6 nu 1/2 ratio was directly proportional to the anisotropy of the ring motions as measured by the ratio of the two correlation times. At any given temperature relative to Tm, the C-3 and C-6 nu 1/2's and the C-3/C-6 nu 1/2 ratios were larger for cholesterol esters which have a cholesteric phase than for esters which have no cholesteric phase, showing that steroid ring motions were more restricted and more anisotropic prior to the formation of a cholesteric phase. Cholesteryl erucate and cholesteryl nervonate have longer regions of FA chain interactions which result in greater chain cooperativity, apparently preventing the preordering of steroid rings to the degree necessary for formation of a cholesteric phase. Thus, these esters form the smectic phase directly from the isotropic liquid. These results are applied to the cholesterol ester transition in plasma low-density lipoproteins.


Acute hepatic steatosis: a helpful diagnostic feature in metallic phosphide-poisoned horses.
Metal phosphides, particularly zinc and aluminum phosphide, occasionally poison horses and other equids following their use as rodenticides and insecticides. Grain-based aluminum phosphide baits are used to control rodents such as prairie dogs. The clinical course in intoxicated horses is short (<24-48 h), and animals may be found dead. Hepatic lesions caused by phosphine poisoning are not well described. Laboratory confirmation depends on detecting phosphine gas in gastric contents. Eight horses and a mule were exposed to zinc phosphide used to control prairie dogs on a Wyoming ranch. Three of 9 exposed equids developed some combination of sweating, ataxia, anxiety, and colic; 2 died acutely, and 1 recovered. A diagnosis of zinc phosphide was made by detecting phosphine in stomach contents from a horse and a mule. The liver was pale and swollen in the affected horse, which died after a clinical course of ~12 h. Other changes were generalized congestion and edema, pulmonary edema, and acute cerebrocortical edema. There was diffuse hepatocellular microvesicular steatosis. Similar histologic lesions were present in 7 equine livers from 2 previously published episodes of metallic phosphide poisoning. Older lesions (>24 h of clinical signs) had centrilobular hepatic necrosis with congestion and a mixture of microvesicular and macrovesicular steatosis. Phosphine poisoning should be considered in horses that die acutely and are found to have steatosis, either with or without hepatocellular necrosis.


Tobacco expenditures and its implications for household resource allocation in Cambodia.
To assess the determinants of smoking behaviour and to estimate the impact of tobacco consumption on the consumption of other commodities by Cambodian households. To assess the determinants of smoking in Cambodia, the authors used a logistic regression model that estimated the probability of an individual smoking, given a set of socioeconomic and demographic characteristics. A Seemingly Unrelated Regression method was used to assess the impact of tobacco consumption on the consumption of other commodities. The nationally representative 2004 Cambodia Socio-Economic Survey, collected by the National Institute of Statistics of the Ministry of Planning in Cambodia, was used for the analysis. Smoking in Cambodia is influenced by a variety of factors such as gender, marital status, age, ethnicity, literacy, health status and perceptions about the health consequences of tobacco use. The authors found that spending on tobacco crowds out expenditures on education and clothing at the national level and expenditures on food for low- and middle-income households. The first analysis of the study showed that increased education is associated with lower daily smoking, and the second analysis revealed that expenditures on tobacco crowds out expenditures on education. Combining these two results points to a vicious circle where low education means higher likelihood of smoking, which in turn results in lower spending on education. Such budget allocation clearly has negative intergenerational consequences.


Population pharmacokinetics of troxacitabine, a novel dioxolane nucleoside analogue.
To develop and validate a population pharmacokinetic model for troxacitabine, a novel l-nucleoside analogue, administered by short infusion; to characterize clinical covariates that influence pharmacokinetic variability; and to design a dosage rate for continuous infusion administration to achieve low micromolar concentrations, which may be more efficacious than shorter infusions. Plasma samples from 111 cancer patients receiving troxacitabine (0.12-12.5 mg/m(2)) as a 30-minute infusion in phase I trials were used to develop the model with NONMEM. Clinical covariates evaluated included creatinine clearance, body surface area, age, and sex. From the model, a troxacitabine dosage rate of 2.0 to 3.0 mg/m(2)/d was expected to achieve a target concentration of 0.1 micromol/L; plasma samples were obtained during the infusion from eight patients receiving troxacitabine as a 3-day infusion. Troxacitabine pharmacokinetics were characterized by a three-compartment linear model. The mean value for systemic clearance [interindividual variability (CV%)] from the covariate-free model was 9.1 L/h (28%). Creatinine clearance and body surface area accounted for 36% of intersubject variation in clearance. Troxacitabine 2.0 mg/m(2)/d (n = 3) and 3.0 mg/m(2)/d (n = 5) for 3 days produced mean +/- SD end of infusion concentrations of 0.12 +/- 0.03 and 0.15 +/- 0.03 micromol/L, respectively. Renal function and body surface area were identified as sources of troxacitabine pharmacokinetic variability. The population pharmacokinetic model model-derived dosage rates for continuous infusion administration successfully achieved predetermined target plasma concentrations. The present model may be used to optimize treatment with troxacitabine by developing a dosing strategy based on both renal function and body size.