Decreased Lamina Cribrosa Beam Thickness and Pore Diameter Relative to Distance From the Central Retinal Vessel Trunk.
To investigate how the lamina cribrosa (LC) microstructure changes with distance from the central retinal vessel trunk (CRVT), and to determine how this change differs in glaucoma. One hundred nineteen eyes (40 healthy, 29 glaucoma suspect, and 50 glaucoma) of 105 subjects were imaged using swept-source optical coherence tomography (OCT). The CRVT was manually delineated at the level of the anterior LC surface. A line was fit to the distribution of LC microstructural parameters and distance from CRVT to measure the gradient (change in LC microstructure per distance from the CRVT) and intercept (LC microstructure near the CRVT). A linear mixed-effects model was used to determine the effect of diagnosis on the gradient and intercept of the LC microstructure with distance from the CRVT. A Kolmogorov-Smirnov test was applied to determine the difference in distribution between the diagnostic categories. The percent of visible LC in all scans was 26 ± 7%. Beam thickness and pore diameter decreased with distance from the CRVT. Glaucoma eyes had a larger decrease in beam thickness (-1.132 ± 0.503 μm, P = 0.028) and pore diameter (-0.913 ± 0.259 μm, P = 0.001) compared with healthy controls per 100 μm from the CRVT. Glaucoma eyes showed increased variability in both beam thickness and pore diameter relative to the distance from the CRVT compared with healthy eyes (P < 0.05). These findings results demonstrate the importance of considering the anatomical location of CRVT in the assessment of the LC, as there is a relationship between the distance from the CRVT and the LC microstructure, which differs between healthy and glaucoma eyes.


The retention of gold crowns on human dentine preparations--a comparison of eight cements.
Experiments were carried out to compare the retentive properties of eight dental luting cements, using gold crowns cemented onto human dentine. The order of retention of the cements was: 1 Composite (Panavia-Ex, J & S Davis); 2 Glass-ionomer. (AquaCem, DeTrey); Glass-ionomer (Ketac-Bond, Cottrell); and Polycarboxylate (Bondalcap, Vivadent); 3 Polycarboxylate (Poly F Plus, DeTrey); Zinc phosphate (DeTrey); and Zinc phosphate (Phosphacap, Vivadent); and 4 Zinc oxide/eugenol, alumina, EBA (Opotow, Teledyne Getz).


Strategies for improving the production of bio-based vanillin.
Vanillin (4-hydroxy-3-methoxybenzaldehyde) is one of the most popular flavors with wide applications in food, fragrance, and pharmaceutical industries. However, the high cost and limited yield of plant extraction failed to meet the vast market demand of natural vanillin. Vanillin biotechnology has emerged as a sustainable and cost-effective alternative to supply vanillin. In this review, we explored recent advances in vanillin biosynthesis and highlighted the potential of vanillin biotechnology. In particular, we addressed key challenges in using microorganisms and provided promising approaches for improving vanillin production with a special focus on chassis development, pathway construction and process optimization. Future directions of vanillin biosynthesis using inexpensive precursors are also thoroughly discussed.


Electrophysiological responses of nucleus tractus solitarius neurons to CCK and gastric distension in newborn lambs.
The effect of cervical vagus nerve stimulation, gastric distension and CCK-8S administration was studied on the activity of 120 neurons located in the nucleus tractus solitarius (NTS) of anesthetized newborn lambs. One hundred cells responded to the three different inputs. The distribution of the cells in the NTS was from 3 mm rostral to 3 mm caudal to the obex, the major responsive cells being located at the level of the obex. Neurons were either excited or inhibited by gastric distension and CCK-8S, and the responses to these two stimuli were always in the same direction. A small number of cells responded to gastric distension and CCK-8S but not to vagus nerve stimulation. Injection of the CCK-A receptor antagonist 2-NAP abolished both the responses to CCK-8S and to gastric distension. The results are consistent with the idea that CCK-8S acts directly on vagal mechanoreceptive endings in the gastric corpus close to duodenum. These results from lambs may reflect the pathway by which gastric distension and peripheral CCK-8S modulate NTS cells activity during colostrum ingestion, which could in turn activate structures related to learning and memory processes involved in the development of mother preference.


Selection of cell fate in the organ of Corti involves the integration of Hes/Hey signaling at the Atoh1 promoter.
Determination of cell fate within the prosensory domain of the developing cochlear duct relies on the temporal and spatial regulation of the bHLH transcription factor Atoh1. Auditory hair cells and supporting cells arise in a wave of differentiation that patterns them into discrete rows mediated by Notch-dependent lateral inhibition. However, the mechanism responsible for selecting sensory cells from within the prosensory competence domain remains poorly understood. We show in mice that rather than being upregulated in rows of cells, Atoh1 is subject to transcriptional activation in groups of prosensory cells, and that highly conserved sites for Hes/Hey repressor binding in the Atoh1 promoter are needed to select the hair cell and supporting cell fate. During perinatal supporting cell transdifferentiation, which is a model of hair cell regeneration, we show that derepression is sufficient to induce Atoh1 expression, suggesting a mechanism for priming the 3' Atoh1 autoregulatory enhancer needed for hair cell expression.


The microbial pan-genome.
A decade after the beginning of the genomic era, the question of how genomics can describe a bacterial species has not been fully addressed. Experimental data have shown that in some species new genes are discovered even after sequencing the genomes of several strains. Mathematical modeling predicts that new genes will be discovered even after sequencing hundreds of genomes per species. Therefore, a bacterial species can be described by its pan-genome, which is composed of a "core genome" containing genes present in all strains, and a "dispensable genome" containing genes present in two or more strains and genes unique to single strains. Given that the number of unique genes is vast, the pan-genome of a bacterial species might be orders of magnitude larger than any single genome.


Isoproterenol inhibits transcription of cardiac cytokine genes induced by reactive oxygen intermediates.
Cytokines such as tumor necrosis factor alpha (TNF-alpha) are produced by the myocardium in heart disease and might be stimulated by reactive oxygen. In some cell types, cyclic adenosine monophosphate (AMP) inhibits TNF-alpha production. The authors tested the hypothesis that stimulation of cardiac beta-adrenergic receptors would inhibit cytokine gene transcription induced by reactive oxygen. Rat hearts were perfused with buffer containing hypoxanthine. Reactive oxygen intermediates were generated by infusion of xanthine oxidase. Myocardial mRNA encoding 11 cytokines was determined. TNF-alpha, interleukin-6, and cyclic AMP were measured in the coronary effluent. In control hearts, of the screened RNA, only mRNA encoding interleukin-1beta, -4, and -6 was detected. Stimulation with hypoxanthine-xanthine oxidase (HX-XO) induced detectable mRNA for TNF-alpha and interleukin-5 and increased mRNA band density for interleukin-1beta, -4, and -6. Simultaneous infusion of isoproterenol inhibited HX-XO-stimulated cytokine gene expression and caused release of cyclic AMP into the coronary effluent. In control hearts, TNF-alpha was not detected in the coronary effluent. After HX-XO administration, TNF-alpha was reliably detected at 60 min and interleukin-6 at 90 min. Simultaneous infusion of isoproterenol inhibited TNF-alpha and interleukin-6 release. Inclusion of propranolol in the perfusion buffer blocked the isoproterenol-induced inhibition of HX-XO-stimulated TNF-alpha release and release of cyclic AMP into the coronary effluent. In addition, elevating myocardial cyclic AMP with forskolin also blocked release of TNF-alpha stimulated by HX-XO. Finally, delaying infusion of isoproterenol until 30 min after HX-XO administration still suppressed release of TNF-alpha. Reactive oxygen species activate cytokine gene transcription in the myocardium. The sympathetic nervous system, acting through beta-receptors to elevate myocardial cyclic AMP, regulates cardiac cytokine production by inhibition of transcription.


A facile template route to periodic mesoporous organosilicas nanospheres with tubular structure by using compressed CO2.
Periodic mesoporous organosilicas (PMOs) nanospheres with tubular structure were prepared with compressed CO2 using cationic and anionic mixed surfactant (CTAB/SDS) and triblock copolymer Pluronic P123 as bi-templates. TEM, N2 adsorption-desorption, solid NMR, and FTIR were employed to characterize the obtained materials. Compressed CO2 severed as acidic reagent to promote the hydrolysis of organosilicas, and could tune the morphology and structure of the obtained PMOs nanomaterials simple by adjusting the CO2 pressure during the synthesis process. Rhodamine B (RB) and Ibuprofen (IBU), as the model dye and drug, were loaded into the prepared nanomaterials to reveal its adsorption and desorption ability. Furthermore, different molars of the surfactant (CTAB/SDS) and organosilane precursor (BTEB) were investigated to show the effect of the surfactant concentration on the morphology and structure of the PMOs prepared with compressed CO2, and some different structures were obtained. A possible mechanism for the synthesis of PMOs with tubular structure using compressed CO2 was proposed based on the experimental results.


Circle chain embracing donor-acceptor organic dye: simultaneous improvement of photocurrent and photovoltage for dye-sensitized solar cells.
We demonstrate for the first time that employing a circle chain embracing π-conjugated backbone is a promising strategy to construct superior organic sensitizers for dye-sensitized solar cells (DSCs), with simultaneous improvement of photocurrent and photovoltage. A DSC based on one circle chain embracing dye produced a high conversion efficiency of 8.34%.


Echocardiographically evaluated site of attachment of atrial myxoma may predict recurrence.
We explore the association between the site of attachment of nonfamilial left atrial myxoma and it's recurrence. Forty-three (11 male and 32 female; mean age, 55.9±13.6 years) of 49 consecutive patients with nonfamilial left atrial myxoma who had been evaluated with preoperative echocardiography, X were available for postoperative follow-up with transthoracic echocardiography, transesophageal echocardiography, or both, for an average period of 85.2±54.2 months (range, 6.5 to 215.5 months). We compared preoperative clinical and echocardiographic features of recurrent and nonrecurrent myxomas. Three (7%) of the 43 cases of atrial myxoma recurred at the same site after 24.1±7.6 months. Involvement of the mitral valve annulus or mitral valve leaflet (3 vs 0,p<0.001) was observed in the recurrent myxomas, but the two groups did not differ significantly in any other clinical features, laboratory data, or echocardiographic features of the recurrent and nonrecurrent myxoma. Preoperative echocardiographic observation of involvement of the mitral valve annulus or mitral valve leaflet may predict recurrence after surgery. Regular follow-up echocardiography was useful in the early detection of recurrence.


Influence of an electron-deficient bridging o-carborane on the electronic properties of an [FeFe] hydrogenase active site model.
The IR carbonyl stretching frequencies of [Fe2(SRS)(CO)6] complexes correlate well with their first reduction potential; an [FeFe] hydrogenase model with a very mild reduction potential has been realized by using a strongly electron deficient carborane-dithiolate bridge.


Neuroadaptive changes in the mesocortical glutamatergic system during chronic nicotine self-administration and after extinction in rats.
Nicotine self-administration causes adaptation in the mesocorticolimbic glutamatergic system, including the up-regulation of ionotropic glutamate receptor subunits. We therefore determined the effects of nicotine self-administration and extinction on NMDA-induced glutamate neurotransmission between the medial prefrontal cortex (mPFC) and ventral tegmental area (VTA). On day 19 of nicotine SA, both regions were microdialyzed for glutamate while mPFC was sequentially perfused with Kreb's Ringer buffer (KRB), 200 microM NMDA, KRB, 500 microM NMDA, KRB, and 100 mM KCl. Basal glutamate levels were unaffected, but nicotine self-administration significantly potentiated mPFC glutamate release to 200 microM NMDA, which was ineffective in controls. Furthermore, in VTA, nicotine self-administration significantly amplified glutamate responses to both mPFC infusions of NMDA. This hyper-responsive glutamate neurotransmission and enhanced glutamate subunit expression were reversed by extinction. Behavioral studies also showed that a microinjection of 2-amino-5-phosphonopentanoic acid (NMDA-R antagonist) into mPFC did not affect nicotine or sucrose self-administration. However, in VTA, NBQX (AMPA-R antagonist) attenuated both nicotine and sucrose self-administration. Collectively, these studies indicate that mesocortical glutamate neurotransmission adapts to chronic nicotine self-administration and VTA AMPA-R may be involved in the maintenance of nicotine self-administration.


Number of adult day care centers increasing, but payment is slow.
Adult day care programs are expanding throughout the United States, but funding has not kept pace with organizational development. Hospitals explores the issues and looks at three successful programs in diverse settings.


Retrograde transport of goldfish optic nerve proteins labeled by N-succinimidyl [3H]propionate.
Following injection of the acylating reagent N-succinimidyl [3H]propionate into the optic nerve of goldfish, labeled protein appeared in the ipsilateral retina and contralateral tectum in a time-dependent manner. Autoradiography indicated the presence of the labeled material in the neuroplasm of the retinal ganglion cells and their projections. While most of the recoverable injected radioactivity was confined to the injection site even after 1 week, labeled proteins arriving in the retina by retrograde flow or in the tectum by anterograde flow had distinctly different patterns, a result suggesting specific transport processes rather than diffusion. In contrast to reported studies with the labeling agent in other species, a prominent 68,000 molecular weight component was not seen. The results are discussed in relation to the role of retrograde transport in regeneration.


Linker histone-DNA complexes: enhanced stability in the presence of aluminum lactate and implications for Alzheimer's disease.
The binding of human brain linker histone proteins to a radiolabelled human Alu repetitive element was examined by mobility shift assay. Analysis of the complexes formed from protein extracts of whole neocortical nuclei, under physiological conditions in vitro revealed that linker histone H1(0) has the highest affinity for the Alu DNA sequence. The linker histone-DNA complexes assembled in the presence of aluminum lactate were more resistant to sodium chloride-induced dissociation than those formed in the presence of sodium lactate. The enhanced stability of deoxyribonucleoprotein (DNP) complexes in the presence of the aluminum cation may be of significance in neurodegenerative conditions such as Alzheimer's disease where aluminum preferentially associates with DNA containing structures of the nucleus.


Pharmacokinetics of naftidrofuryl in patients with renal impairment.
Naftidrofuryl (CAS 31329-57-4) is used, mainly in elderly patients, in the treatment of various vascular disorders. The aim of this study was to evaluate and compare the pharmacokinetics of naftidrofuryl after single oral administration of a 200 mg naftidrofuryl tablet (Praxilene) in caucasian male and female subjects with renal impairment versus healthy volunteers. This prospective and open study was conducted in three parallel groups: Group A = healthy subjects with a Cl(CR) > 80 ml/min, Group B = uraemic patients with a 20 < or = Cl(CR) < 40 ml/min, Group C = uraemic patients with a Cl(CR) < 20 ml/min. Blood samples were taken over a period of 32 h after dosing. The mean values (+/-SD) of the pharmacokinetic parameters of naftidrofuryl for group A were as follows: tmax: 1.3 h (median), Cmax: 174 +/- 46 ng/ml, t(1/2 beta): 4.4 +/- 1.1 h, AUC(0-infinity): 1541 +/- 384 ng x h/ml; for group B: tmax: 2.5 h (median), Cmax: 239 +/- 94 ng/ml, t(1/2 beta): 5.0 +/- 1.2 h, AUC(0-infinity): 2361 +/- 751 ng x h/ml; for group C: tmax: 3.0 h (median), Cmax: 236 +/- 104 ng/ml, t(1/2 beta): 5.0 +/- 2.1 h, AUC(0-infinity): 2488 +/- 2003 ng x h/ml. The statistical analysis was performed on the pharmacokinetic parameters with one-way ANOVA in order to compare each group. No significant difference between each group was observed. In conclusion, renal insufficiency did not appear to influence the pharmacokinetic profile of oral naftidrofuryl.


Determination of sunset yellow in soft drinks based on fluorescence quenching of carbon dots.
Fluorescent carbon dots was prepared by heating N-(2-hydroxyethyl)ethylene diaminetriacetic acid in air. The carbon dots were not only highly soluble in water but also uniform in size, and possessed strong blue fluorescence and excitation wavelength-dependent emission properties with the maximum excitation and emission wavelength at 366nm and 423nm, respectively. Food colorant sunset yellow whose excitation and emission wavelength at 303nm and 430nm could selectively quench the fluorescence of carbon dots, efficient fluorescent resonance energy transfer between the carbon dots and sunset yellow is achieved. This was exploited to design a method for the determination of sunset yellow in the concentration range from 0.3 to 8.0μmolL(-1), with a limit of detection (3σ/k) of 79.6nmolL(-1). Furthermore the fluorimetric detection method was established and validated for sunset yellow in soft drinks samples with satisfactory results.


Modification of gamma ray induced changes in the mouse hepatocytes by Centella asiatica extract: in vivo studies.
Ionizing radiation, besides causing direct damage also generates reactive oxygen species that are capable of inducing damage to various organs. Pretreatment with Centella asiatica 1 h prior to irradiation at a dose rate of 100 mg/kg body weight was found to be effective against radiation induced damage in the liver. The number of normal hepatocytes was higher in the Centella asiatica pretreated group in comparison with the irradiated only group. The number of binucleated cells and abnormal hepatocytes was less in comparison with the animals irradiated without Centella asiatica pretreatment.


Power boosting the grafts in hair transplantation surgery. Evaluation of a new storage medium.
When performing hair transplantation procedures, it is of the foremost importance to try to obtain the maximum survival rate possible of transplanted micrografts. Aim of this study was to evaluate, in an in vitro model, the effects of preserving micrografts, for five hours, in an enriched storage medium in order to enhance the survival rate of hair micrografts. A total of 200 human anagen hair follicles was obtained from ten male patients. Follicles were thus randomly assigned to one of the following group: Group A (control; n = 100 follicles), preserved for five hours in saline, and Group B (experimental; n = 100 follicles), preserved for five hours in a storage medium, containing adenosine triphosphate-magnesium chloride and deferoxamine mesylate. Isolated hair follicles from both Groups were then cultured for 10 days. A statistically significant difference was found between the survival rate of experimental (98%) and control follicles (87%). In our opinion, a "metabolic preconditioning" of micrografts by means of storing them for 5 hours in the described medium may be of some utility in augmenting the survival rate of hair grafts when performing hair transplantation surgery.


Study into the role of cholecystokinin in bombesin-stimulated pancreatic growth in rats and hamsters.
The effects of cholecystokinin and bombesin on growth of the exocrine pancreas have been studied extensively in rats but not in hamsters. Since hamsters are frequently used for studying pancreatic carcinogenesis it seems highly relevant to determine the effects of these peptides on growth of the hamster pancreas as well. In order to determine whether or not bombesin stimulates pancreatic growth in hamsters and to investigate the role of cholecystokinin in mediating this effect, we conducted a 2-week experiment in which cholecystokinin and bombesin were administered to both rats and hamsters, either with or without lorglumide (CR-1409; a specific cholecystokinin receptor antagonist). Rats were included in the study for comparison. Cholecystokinin and bombesin were found to stimulate pancreatic growth and DNA synthesis in both species. Lorglumide did not significantly influence the effect of bombesin on pancreatic weight but did significantly inhibit DNA synthesis in both species. The effect of cholecystokinin on pancreatic weight was greater in hamsters than in rats. This effect was significantly inhibited by lorglumide in hamsters but not in rats, whereas a significant decrease in DNA content was attained in both species. Thus, this study shows marked differences between rats and hamsters in the pancreatic growth response to cholecystokinin and bombesin.


Surface microanalysis with slow electrons.
Microanalysis on the 10-nm level using imaging, diffraction, and spectroscopy of slow photo-emitted and reflected electrons is discussed. The instrumentation that uses a cathode lens is briefly reviewed, and a number of applications illustrate the power of this microanalysis method.


Quality information disclosure and health insurance demand: evidence from VA hospital report cards.
This study examines the effect of public reporting of quality information on the demand for public insurance. In particular, we examine the effect of the introduction of Veterans Affairs (VA) hospital quality report cards in 2008. Using data from the Current Population Survey in 2005-2015, we find that new information about the quality of a VA hospital had a significant effect on VA coverage among veterans living in the same Metropolitan Statistical Area (MSA). Despite the significant effect on VA coverage, the quality report did not have a spillover effect on veterans' labor supply. Moreover, updated quality information released in later years, which was presented in a less straightforward form, led to no additional changes in VA coverage. These findings suggest that quality reports for public insurance programs can be used as a policy lever to facilitate take up decision among potential beneficiaries.


Subversion of early innate antiviral responses during antibody-dependent enhancement of Dengue virus infection induces severe disease in immunocompetent mice.
Dengue is a mosquito-borne disease caused by one of four serotypes of Dengue virus (DENV-1-4). Epidemiologic and observational studies demonstrate that the majority of severe dengue cases, dengue hemorrhagic fever and dengue shock syndrome (DHF/DSS), occurs predominantly in either individuals with cross-reactive immunity following a secondary heterologous infection or in infants with primary DENV infections born from dengue-immune mothers, suggesting that B-cell-mediated and antibody responses impact on disease evolution. We demonstrate here that B cells play a pivotal role in host responses against primary DENV infection in mice. After infection, μMT(-/-) mice showed increased viral loads followed by severe disease manifestation characterized by intense thrombocytopenia, hemoconcentration, cytokine production and massive liver damage that culminated in death. In addition, we show that poly and monoclonal anti-DENV-specific antibodies can sufficiently increase viral replication through a suppression of early innate antiviral responses and enhance disease manifestation, so that a mostly non-lethal illness becomes a fatal disease resembling human DHF/DSS. Finally, treatment with intravenous immunoglobulin containing anti-DENV antibodies confirmed the potential enhancing capacity of subneutralizing antibodies to mediate virus infection and replication and induce severe disease manifestation of DENV-infected mice. Thus, our results show that humoral responses unleashed during DENV infections can exert protective or pathological outcomes and provide insight into the pathogenesis of this important human pathogen.


Liver surgery in the 2020s: ante-situm and in-situ resection are still indicated - A single-center study.
Tumor infiltration of the hepatic outflow comprising all three hepatic veins and the inferior vena cava remains a surgical challenge. Liver resection under total vascular exclusion with or without extracorporeal bypass has been described as a therapeutic option for these tumors. Here, we present our experience with these complex surgical methods. We searched our database for patients treated with an in-situ or ante-situm liver resection (ISR and ASR, respectively) with extracorporeal bypass. We collected demographic and perioperative data. From January 2010 to December 2021, we performed 2122 liver resections. Nine patients were treated with ASR and five were treated with ISR. Out of these 14 patients, six had colorectal liver metastases, six had cholangiocarcinoma, and two had non-colorectal liver metastases. The median operative time and bypass time in all patients were 536.5 and 150 min, respectively. Compared with ISR, ASR required a longer operative time (ASR 586 min and ISR 495 min) and a longer bypass time (ASR 155 min and ISR 122 min). Morbidity (Clavien-Dindo grade > 3A adverse events) occurred in 78.5% of patients. 90-day postoperative mortality was 7%. Median overall survival was 33 months. Seven patients experienced recurrence. In these patients, median disease-free survival was 9 months. Resection of tumors infiltrating the hepatic outflow poses a high risk for patients. However, with rigorous selection and an experienced perioperative team, these patients can be treated surgically with reasonable oncological outcomes.


A new role for the cellular PABP repressor Paip2 as an innate restriction factor capable of limiting productive cytomegalovirus replication.
The capacity of polyadenylate-binding protein PABPC1 (PABP1) to stimulate translation is regulated by its repressor, Paip2. Paradoxically, while PABP accumulation promotes human cytomegalovirus (HCMV) protein synthesis, we show that this is accompanied by an analogous increase in the abundance of Paip2 and EDD1, an E3 ubiquitin ligase that destabilizes Paip2. Coordinate control of PABP1, Paip2, and EDD1 required the virus-encoded UL38 mTORC1 activator and resulted in augmented Paip2 synthesis, stability, and association with PABP1. Paip2 synthesis also increased following serum stimulation of uninfected normal fibroblasts, suggesting that this coregulation may play a role in how uninfected cells respond to stress. Significantly, Paip2 accumulation was dependent on PABP accrual, as preventing PABP1 accumulation suppressed viral replication and inhibited the corresponding Paip2 increase. Furthermore, depleting Paip2 restored the ability of infected cells to assemble the translation initiation factor eIF4F, promoting viral protein synthesis and replication without increasing PABP1. This establishes a new role for the cellular PABP1 inhibitor Paip2 as an innate defense that restricts viral protein synthesis and replication. Moreover, it illustrates how a stress-induced rise in PABP1 triggered by virus infection can counter and surpass a corresponding increase in Paip2 abundance and stability.


Intravenous immunoglobulin therapy in systemic lupus erythematosus-associated thrombocytopenia.
Seven patients with thrombocytopenia and systemic lupus erythematosus were treated with intravenous (IV) doses of human immunoglobulin to assess clinical response and to examine the mechanism of action of IV immunoglobulin in these patients. Five of 7 patients had a greater than 50% increase in their platelet counts. Four of these patients had a sustained benefit of at least 6 months duration. The initial effectiveness of IV immunoglobulin therapy was not dependent on the reduction of levels of circulating platelet-binding IgG or circulating immune complexes.


Expression of cyclin dependent kinase inhibitor p21waf1/cip1 in premalignant and malignant oral lesions: relationship with p53 status.
p21waf1/cip1 protein, an inhibitor of cyclin dependent kinases, is a critical downstream target in the p53-specific pathway of growth control, and can also be induced by p53 independent pathways in relation to terminal differentiation. p21waf1 is also a putative tumour suppressor. Hence, we sought to determine whether this protein is abnormally expressed during betel- and tobacco-related oral oncogenesis. The aim was to determine whether a correlation exists between the expression profile of p21 and clinicopathological parameters of the patients, as well as with their p53 status. Immunohistochemical analysis showed that the expression of p21 protein in premalignant lesions was consistently elevated in the superficial, differentiated cells of the epithelium, while overexpression of the p53 tumour suppressor gene was observed in the basal proliferating layers of the epithelium. Our study demonstrated that p21 overexpression is associated with differentiation in proliferating dysplasias and squamous cell carcinomas (SCCs). The expression of p21 and p53 proteins was observed in 11/25 premalignant lesions. In 7 of these 11 cases, a heterogenous pattern of expression of p21 and p53 was observed. Four of these 11 premalignant and 30/51 malignant lesions showed concordant expression of both p21 and p53 proteins. The discordant p21 +/p53- phenotype was observed in 4/25 premalignant lesions and 5/51 oral SCCs. The p21-/p53+ phenotype was observed in 5/25 premalignant lesions and 7/51 oral SCCs. These results suggest that induction of p21 occurs by both p53 dependent and independent mechanisms during oral tumorigenesis.


Analysis of left ventricular wall motion based on volumetric deformable models and MRI-SPAMM.
We present a new approach for the analysis of the left ventricular shape and motion based on the development of a new class of volumetric deformable models. We estimate the deformation and complex motion of the left ventricle (LV) in terms of a few parameters that are functions and whose values vary locally across the LV. These parameters capture the radial and longitudinal contraction, the axial twisting, and the long-axis deformation. Using Lagrangian dynamics and finite-element theory, we convert these volumetric primitives into dynamic models that deform due to forces exerted by the datapoints. We present experiments where we used magnetic tagging (MRI-SPAMM) to acquire datapoints from the LV during systole. By applying our method to MRI-SPAMM datapoints, we were able to characterize the 3-D shape and motion of the LV both locally and globally, in a clinically useful way. In addition, based on the model parameters we were able to extract quantitative differences between normal and abnormal hearts and visualize them in a way that is useful to physicians.


Three-dimensional sonography of the placental and uterine spiral vasculature: influence of maternal age and parity.
: To test the influence of maternal age and parity on placental volume and uterine spiral vasculature volume, their mean gray value, and 3-dimensional power Doppler (3DPD) indices in normal pregnancy. : Women with a gestational age of 14-25 weeks were included in the study. Exclusion criteria included posterior placenta, structural or chromosomal anomaly, pregnancy complications, and unknown pregnancy outcome. 3D sonograms of the placenta and uterine spiral vasculature were obtained. Virtual Organ Computer-aided AnaLysis imaging software was used to calculate the volumes, mean gray value, and 3DPD indices. : The study included 199 women, the majority of whom were Filipino, Japanese, or part Hawaiian. The placental volume, uterine spiral vasculature volume, mean gray value, and 3DPD indices were similar in all women regardless of maternal age. However, uterine spiral vasculature volume was significantly larger in women younger than 25 years, compared with women 35 years of age and older. Parity influenced all placental 3DPD indices, whereas placental volume, uterine spiral vasculature volume, mean gray value, and 3DPD indices remained unaffected. : Maternal age influenced uterine spiral vasculature volume, whereas parity influenced all placental 3DPD indices.


miR-223 overexpression inhibits doxorubicin-induced autophagy by targeting FOXO3a and reverses chemoresistance in hepatocellular carcinoma cells.
Doxorubicin is conventionally used in chemotherapy against hepatocellular carcinoma (HCC), but acquired resistance developed during long-term therapy limits its benefits. Autophagy, a conserved catabolic process for cellular self-protection and adaptation to the changing environment, is regarded as a potential clinical target to overcome doxorubicin resistance. In this study, the potential role of miR-223 in modulating doxorubicin-induced autophagy and sensitivity were evaluated in four transfected human HCC cell lines, and the in vivo relevance was assessed using a mouse xenograft model of HCC. We found that the well-defined miR-223 is expressed at low levels in doxorubicin treated HCC cells and that miR-223 overexpression inhibits the doxorubicin-induced autophagy that contributes to chemoresistance. Blockade of autophagic flux by chloroquine resulted in the failure of miR-223 inhibitor to suppress doxorubicin sensitivity of HCC cells. We further identified FOXO3a as a direct downstream target of miR-223 and primary mediator of the regulatory effect of miR-223 on doxorubicin-induced autophagy and chemoresistance in HCC cells. Finally, we confirmed the enhancement of doxorubicin sensitivity by agomiR-223 in xenograft models of HCC. These findings establish a novel miRNA-based approach for autophagy interference to reverse doxorubicin resistance in future chemotherapy regimens against human HCC.


Development and psychometric testing of the medication taking behavior tool in Thai patients.
The previous Thai version of the 8-item Morisky Medication Adherence Scale (8-item MMAS) showed poor sensitivity and unacceptable internal consistency reliability. To develop and test the psychometric properties of a new medication taking behavior measure for Thai patients (MTB-Thai) including practicality, reliability and validity. This study was conducted with adult outpatients regularly taking any medicines for at least three months from three university hospitals in Bangkok, Thailand, between July 2014 and March 2015. The study was approved by the Ethical Committees of the three hospitals. Practicality was assessed by administration time and the percentage of missing data. Internal consistency and test-retest reliability were evaluated employing Cronbach's alpha and intraclass correlation coefficients (ICCs), respectively. Validity was evaluated with content, construct, convergent and known-groups validity. Of 1156 patients, the 6-item MTB-Thai had an average administration time of 2 min and no missing data. It showed good Cronbach's alpha value of 0.76 and excellent ICCs of 0.83. The MTB-Thai showed good content validity with the item level and scale level of content validity indexes greater than the acceptable levels of 0.8 and 0.9, respectively. Both exploratory and confirmatory factor analyses showed the MTB-Thai had two domains including unintentional and intentional domains. The MTB-Thai correlated well with the overall medication adherence scale with a Pearson's correlation coefficient of 0.62 (p < 0.001). The sensitivity, specificity, positive predictive value and negative predictive value of the MTB-Thai were 76, 35, 55 and 57 %, respectively. The MTB-Thai was practical, reliable and valid in assessing mediation taking behaviors in Thai patients with chronic diseases.


Hypoglycemic effect of cotton seed aqueous extract in alloxan-induced diabetes mellitus in rats.
The aqueous extract of cotton seed is able to reduce blood sugar in alloxan-induced diabetes mellitus in rats. A dose of 1000 mg/kg was found to be an effective dose. Cotton seed extract was able to enhance the liver glycogen, like glibenclamid, and was also able to reduce blood cholesterol which was found raised in the diabetic state. Further it was able to normalize the altered level in the liver lipid peroxide content. The role of cotton seed aqueous extract is suggested in the lipid metabolism which is altered during diabetes mellitus.


Distribution of substance P immunoreactive nerve terminals within the nucleus tractus solitarius of the rat.
The relative density of the substance P (SP) immunoreactive nerve terminals in the subnuclei of the nucleus tractus solitarius (NTS) of the rat was examined by immunohistochemical light and electron microscopy. A higher density of SP immunoreactive terminals was observed in the medial and dorsolateral subnuclei of the rostral levels to the obex, while SP immunoreactive terminals were only sparsely found in the medial, ventrolateral and interstitial subnuclei from levels 1 mm rostral to 0.5 mm caudal to the obex. Most other parts of the NTS were moderately innervated by SP immunoreactive nerve terminals.


Viral transduction of primary Schwann cells using a Cre-lox system to regulate GDNF expression.
Glial cell-line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor known to enhance motor nerve regeneration following its delivery. However, recent studies have determined that extended GDNF delivery to regenerating axons can entrap motor axons at the site of GDNF delivery. This entrapment leads to reduced motor axons available to reinnervate muscle. To address this issue, we designed a cell-based GDNF expression system that can temporally regulate protein expression using an inducible gene excision mechanism to prevent entrapment at the site of expression. To design this system for regulation of GDNF expression, we transduced two lentiviral vectors, one containing a constitutively active GDNF transgene flanked by two loxP sites, and the other containing a tetracycline-inducible cre transgene along with its constitutively active transactivator, into Schwann cells (SCs). These SCs over-express GDNF, but expression can be suppressed through the administration of tetracycline family antibiotics, such as doxycycline. The engineered SCs produced significantly more GDNF as compared to untransduced controls, as measured by enzyme-linked immunosorbent assay (ELISA). Following doxycycline treatment, these SCs produced significantly lower levels of GDNF and induced less neurite extension as compared to untreated SCs. Engineered SCs treated with doxycycline showed a marked increase in Cre recombinase expression, as visualized by immunohistochemistry (IHC), providing evidence of a mechanism for the observed changes in GDNF expression levels and biological activity. This cell-based GDNF expression system could have potential for future in vivo studies to provide a temporally controlled GDNF source to promote axon growth.


Severe Intracranial Hypotension After a Middle Fossa Approach for Spontaneous Otogenic Pneumocephalus.
Spontaneous otogenic pneumocephalus is a rare condition caused by erosion of the tegmen tympani with subsequent entrance of air into the intracranial space. Its pathogenesis is thought to involve a previous state of intracranial hypotension, which pulls air into the intracranial cavity. The surgical management involves obliteration of the tegmen defect via a mastoidectomy or a middle fossa approach. Lumbar drainage has been used safely as an adjunct to middle fossa approaches so as to provide brain relaxation and decrease the incidence of postoperative cerebrospinal fluid leaks. A 66-year-old male patient with otogenic pneumocephalus caused by nose blowing underwent repair of a tegmen tympani defect through a middle fossa approach, with the aid of intraoperative lumbar drainage. Progressive neurologic deterioration was seen postoperatively with obtundation and anisocoria. Computed tomography scans of the head demonstrated marked midline shift and transtentorial herniation. Recumbency and blood patch failed to improve the neurological condition. Intrathecal infusion of normal saline enabled clinical and radiological improvement. Intraoperative lumbar drainage during a middle fossa approach for spontaneous otogenic pneumocephalus may dramatically aggravate a state of preexisting intracranial hypotension and lead to transtentorial herniation.


Surface scattering mechanisms of tantalum nitride thin film resistor.
In this letter, we utilize an electrical analysis method to develop a TaN thin film resistor with a stricter spec and near-zero temperature coefficient of resistance (TCR) for car-used electronic applications. Simultaneously, we also propose a physical mechanism mode to explain the origin of near-zero TCR for the TaN thin film resistor (TFR). Through current fitting, the carrier conduction mechanism of the TaN TFR changes from hopping to surface scattering and finally to ohmic conduction for different TaN TFRs with different TaN microstructures. Experimental data of current-voltage measurement under successive increasing temperature confirm the conduction mechanism transition. A model of TaN grain boundary isolation ability is eventually proposed to influence the carrier transport in the TaN thin film resistor, which causes different current conduction mechanisms.


Barriers to Implementation of Rapid and Point-of-Care Tests for Human Immunodeficiency Virus Infection: Findings From a Systematic Review (1996-2014).
Implementation of human immunodeficiency virus rapid and point-of-care tests (RDT/POCT) is understood to be impeded by many different factors that operate at 4 main levels-test devices, patients, providers, and health systems-yet a knowledge gap exists of how they act and interact to impede implementation. To fill this gap, and with a view to improving the quality of implementation, we conducted a systematic review. Five databases were searched, 16,672 citations were retrieved, and data were abstracted on 132 studies by 2 reviewers. Across 3 levels (ie, patients, providers, and health systems), a majority (59%, 112/190) of the 190 barriers were related to the integration of RDT/POCT, followed by test-device-related concern (ie, accuracy) at 41% (78/190). At the patient level, a lack of awareness about tests (15/54, 28%) and time taken to test (12/54, 22%) dominated. At the provider and health system levels, integration of RDT/POCT in clinical workflows (7/24, 29%) and within hospitals (21/34, 62%) prevailed. Accuracy (57/78, 73%) was dominant only at the device level. Integration barriers dominated the findings followed by test accuracy. Although accuracy has improved during the years, an ideal implementation could be achieved by improving the integration of RDT/POCT within clinics, hospitals, and health systems, with clear protocols, training on quality assurance and control, clear communication, and linkage plans to improve health outcomes of patients. This finding is pertinent for a future envisioned implementation and global scale-up of RDT/POCT-based initiatives.


A multigene deletion in the immunoglobulin heavy chain region in a highly atopic individual.
Highly atopic individuals, with marked allergy, have extremely elevated total plasma IgE levels. To determine if atopy could be associated with structural alterations involving the IGHE gene of the immunoglobulin heavy chain constant region, the genomic DNA from five atopic individuals was examined. We describe here the identification of a deletion of approximately 120kb, including the IGHA1, IGHGP, IGHG2, AGHG4, and IGHE genes of the IGH constant region, in one atopic patient. This deletion arose de novo from a maternally derived chromosome. The deletion, although apparently not the primary cause of the atopic phenotype of this patient, could be indirectly responsible for the phenotype by exposing aberrant immunoglobulin-regulating elements within the paternally derived IGH constant region.


A technique for digital alignment of cross-articulated casts in a computer-aided design software program for complete-mouth rehabilitation.
With the advent of computer-aided design and computer-aided manufacturing (CAD-CAM), dental restorations are no longer fabricated in an entirely analog fashion. Successful execution of complete-mouth rehabilitations requires articulation of definitive maxillary and mandibular casts with tooth preparations, as well as cross-articulation with the patient's interim restorations. These cross-articulated records serve as a guide for the dental laboratory technician in fabricating the definitive restorations. In the CAD-CAM workflow, cross-articulation and overlaying of a patient's interim casts over the definitive casts of tooth preparations poses some challenges because of loss of common anatomic data points such as incisive papilla, rugae, and the palate. This article describes a straightforward technique to overcome this common challenge, where a dental laboratory technician can predictably overlay the digitized interim casts to allow for cross-articulation and proceed with designing and fabricating the definitive restorations.


Nondestructive microbial discrimination using single-cell Raman spectra and random forest machine learning algorithm.
Raman microspectroscopy is a powerful tool for obtaining biomolecular information from single microbial cells in a nondestructive manner. Here, we detail steps to discriminate prokaryotic species using single-cell Raman spectra acquisitions followed by data preprocessing and random forest model tuning. In addition, we describe the steps required to evaluate the model. This protocol requires minimal preprocessing of Raman spectral data, making it accessible to non-spectroscopists, yet allows intuitive visualization of feature importance. For complete details on the use and execution of this protocol, please refer to Kanno et al. (2021).


Open-label pilot study of ethosuximide as adjunctive therapy for relieving abdominal pain related to Irritable Bowel Syndrome.
There is clear evidence for an association between irritable bowel syndrome (IBS) and visceral hypersensitivity. This clinical study aimed to assess the adjunct role of ethosuximide, an antiepileptic drug with T-type calcium channel blocking activity, in the relieving of IBS-related abdominal pain. This is a prospective, 3-month, randomized and controlled study of parallel groups. Fifty outpatients who met the inclusion criteria participated in the trial. Patients were allocated randomly: 25 received mebeverine 135 mg three times daily (t.i.d), whereas the other 25 received mebeverine 135 mg t.i.d and ethosuximide 500 mg t.i.d. At baseline and 12 weeks after starting the drug, patients were evaluated by a gastroenterologist. Serum tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-8 (IL-8), faecal myeloperoxidase and faecal neutrophile gelatinase-associated lipocalin (NGAL) levels were tested before and after treatment. The Numeric Pain Rating Scale (NRS) was assessed before and after three months of therapy. After 12 weeks, the ethosuximide group showed a statistically and significantly greater reduction in the serum levels of TNF-α, IL-6, IL-8, faecal myeloperoxidase and faecal NGAL in comparison with the control group after the treatment. Moreover, the ethosuximide group showed a statistically significant decrease in NRS compared with the mebeverine group. Ethosuximide could be a promising adjunct to antispasmodics in the treatment of IBS patients. Trial registration identifier: NCT04217733.


Could chatbots help devise the next pandemic virus?
An MIT class exercise suggests AI tools can be used to order a bioweapon, but some are skeptical.


CT phenotypes in mild-to-moderate chronic obstructive pulmonary disease: difference before and after the age of 60 years.
To investigate the pulmonary phenotype of mild-to-moderate chronic obstructive pulmonary disease (COPD) using quantitative computed tomography CT analysis techniques. Sixty-three patients with stable-phase mild-to-moderate COPD and 78 healthy controls, categorised as those aged <60 (28 and 40 patients, respectively) and ≥60 years (35 and 38, respectively), underwent chest low-dose respiratory dual-phase CT and pulmonary function test. Automatic software was used to measure the proximal airway parameters, and the emphysema and air-trapping indices were recorded. The intergroup differences in each parameter were assessed by one-way analysis of variance. At <60 years of age, the mean Pi10WA (Wall area of a hypothetical airway with an internal perimeter of 10 mm) was greater in the mild-to-moderate COPD group than in the healthy control group (17.04 ± 1.63 versus 16.03 ± 1.16 mm2; p=0.004). Mild or moderate air-trapping was observed in the mild-to-moderate COPD group aged <60 years. There was no significant difference in the proximal airway parameters and inspiratory VI-950 (percent voxels less than -950 HU) between the two groups at age ≥60 years (all p>0.05); however, the expiratory VI-856 (percent voxels less than -856 HU) and mean lung density expiratory/inspiratory ratio (MLDE/I) were higher in the mild-to-moderate COPD group than those in the healthy control group (26.02 [30.23] versus 6.45 [11.16]; 0.88 ± 0.05 versus 0.84 ± 0.04; p<0.001 and p=0.024). For patients with mild-to-moderate COPD, the CT phenotype was predominantly the "airway remodelling" type at <60 years of age, and the "air-trapping" type at ≥60 years of age. Thus, pulmonary CT phenotypes of mild-to-moderate COPD patients of different age groups are different.


Detection of Anaplasma bovis and Anaplasma phagocytophilum from cattle on Yonaguni Island, Okinawa, Japan.
DNA fragments of Anaplasma bovis and Anaplasma phagocytophilum were detected in cattle on Yonaguni Island, Okinawa, Japan. Although the pathogenesis and epidemiology of these pathogens have not yet been clarified, this is the first detection of their presence in cattle from Japan.


Obstructed gall-bladder causing false. Positive liver scanning.
A case is reported in which obstructed gallbladder resulted in a large defect in the liver scan. Causes of false positive liver scans are discussed.


Numerical analysis of pulsatile blood flow and vessel wall mechanics in different degrees of stenoses.
Hemodynamics factors and biomechanical forces play key roles in atherogenesis, plaque development and final rupture. In this paper, we investigated the flow field and stress field for different degrees of stenoses under physiological conditions. Disease is modelled as axisymmetric cosine shape stenoses with varying diameter reductions of 30%, 50% and 70%, respectively. A simulation model which incorporates fluid-structure interaction, a turbulence model and realistic boundary conditions has been developed. The results show that wall motion is constrained at the throat by 60% for the 30% stenosis and 85% for the 50% stenosis; while for the 70% stenosis, wall motion at the throat is negligible through the whole cycle. Peak velocity at the throat varies from 1.47 m/s in the 30% stenosis to 3.2m/s in the 70% stenosis against a value of 0.78 m/s in healthy arteries. Peak wall shear stress values greater than 100 Pa were found for > or =50% stenoses, which in vivo could lead to endothelial stripping. Maximum circumferential stress was found at the shoulders of plaques. The results from this investigation suggest that severe stenoses inhibit wall motion, resulting in higher blood velocities and higher peak wall shear stress, and localization of hoop stress. These factors may contribute to further development and rupture of plaques.


Superspiralization of chiral strings.
A phenomenon of superspiralization was revealed in homochiral solutions of biomimetics with strings lengths from 10(2) nm to 10(2) μ and longer: strings of greater size spontaneously formed in solution are twined of the smaller strings, which also have a helical structure. The chiral pitch depends on the conditions of formation of a particular string and can vary by several times in different strings of the same specimen.


Bmi1 Regulates IκBα Degradation via Association with the SCF Complex.
Bmi1 is a polycomb group protein and regulator that stabilizes the ubiquitination complex PRC1 in the nucleus with no evidently direct link to the NF-κB pathway. In this study, we report a novel function of Bmi1: its regulation of IκBα ubiquitination in the cytoplasm. A deficiency of Bmi1 inhibited NF-κB-mediated gene expression in vitro and a NF-κB-mediated mouse model of arthritis in vivo. Mechanistic analysis showed that Bmi1 associated with the SCF ubiquitination complex via its N terminus and with phosphorylation by an IKKα/β-dependent pathway, leading to the ubiquitination of IκBα. These effects on NF-κB-related inflammation suggest Bmi1 in the SCF complex is a potential therapeutic target for various diseases and disorders, including autoimmune diseases.


Liver transplantation for iatrogenic bile duct injury: a systematic review.
Bile duct injury (BDI) is an infrequent but serious complication of cholecystectomy, often with life-changing consequences. Liver transplantation (LT) may be required following severe BDI, however given the rarity, few large studies exist to guide management for complex BDI. A systematic review was performed to assess post-operative complications, 30-day mortality, retransplant rate and 1-year and 5-year survival following LT for BDI in Medline, EMBASE, Web of Science or Cochrane Clinical Trials Database. Seven articles met inclusion criteria, describing 179 patients that underwent LT for BDI. Secondary biliary cirrhosis (SBC) was the main indication for LT (82.2% of patients). Median model for end-stage liver disease (MELD) scores at time of LT ranged from 16 to 20.5. Median 30-day mortality was 20.0%. The 1-year and 5-year survival ranges were 55.0-84.3% and 30.0-83.3% respectively, and the overall retransplant rate was 11.5%. BDI is rarely indicated for LT, predominantly for SBC following multiple prior interventions. MELD scores poorly reflect underlying morbidity, and exception criteria for waitlisting may avoid prolonged LT waiting times. 30-day mortality was higher than for non-BDI indications, with comparable long term survival, suggesting that LT remains a viable but high risk salvage option for severe BDI.


Laser scanning microscopy approach for semiquantitation of in vitro dermal particle penetration.
Skin penetration of nanoparticles is a recent research area in focus for the aim of development of topical nanoparticulate delivery systems as well as for health risk analysis. So far, monitoring skin penetration of nanoparticles is mostly based on qualitative microscopical examination. Here, we describe an experimental approach for extracting semiquantitative data from multiphoton images of skin specimens treated with gold nanoparticles. This will aid in depicting the factors responsible for enhancing or limiting nanoparticle penetration through the skin barrier.


On the quantitation of SV40 virus by plaque assay and immunoperoxidase technique.
Procedures are described for the quantitation of SV40 virus infectivity by plaque formation within 7 days and T antigen assay by the sensitive and economical indirect immunoperoxidase technique.


Monocyte esterase? A factor involved in the pathogenesis of lymphoproliferative neoplasia.
Monocyte esterase activity was studied in 1,000 doctor-attending patients with normal hematological indices and in 56 patients with non-Hodgkin's lymphoma (NHL) or B chronic lymphocytic leukemia (CLL). The incidence of esterase deficiency was significantly greater in the NHL-CLL patients (7.1%) than in the population group (1.7%; p less than 0.05). In the NHL-CLL group, study of the families showed the esterase deficiency to be a familial characteristic. We postulate that the presence of the anomaly may be either a factor predisposing to the development of the NHL or CLL or a factor indicating a predisposition to these disorders.


Metastatic undifferentiated pleomorphic sarcoma diagnosed by endoscopic ultrasound-guided fine-needle aspiration.
Pathological differentiation is important for suspected lesions of metastatic undifferentiated pleomorphic sarcoma (UPS) because no reliable imaging criteria exist for this entity yet. In the present case, transgastric endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) for the pancreatic tumor and transcolonic EUS-FNA for the intraabdominal tumor contributed to the definitive diagnosis of metastatic UPS, leading to appropriate treatment selection.


Detection of neurophysiological features in female R255X MeCP2 mutation mice.
Rett syndrome (RTT) is a severe neurodevelopmental disorder (NDD) that is nearly always caused by loss of function mutations in Methyl-CpG-binding Protein 2 (MECP2) and shares many clinical features with other NDD. Genetic restoration of Mecp2 in symptomatic mice lacking MeCP2 expression can reverse symptoms, providing hope that disease modifying therapies can be identified for RTT. Effective and rapid clinical trial completion relies on well-defined clinical outcome measures and robust biomarkers of treatment responses. Studies on other NDD have found evidence of differences in neurophysiological measures that correlate with disease severity. However, currently there are no well-validated biomarkers in RTT to predict disease prognosis or treatment responses. To address this, we characterized neurophysiological features in a mouse model of RTT containing a knock-in nonsense mutation (p.R255X) in the Mecp2 locus. We found a variety of changes in heterozygous female Mecp2R255X/X mice including age-related changes in sleep/wake architecture, alterations in baseline EEG power, increased incidence of spontaneous epileptiform discharges, and changes in auditory evoked potentials. Furthermore, we identified association of some neurophysiological features with disease severity. These findings provide a set of potential non-invasive and translatable biomarkers that can be utilized in preclinical therapy trials in animal models of RTT and eventually within the context of clinical trials.


Identification and characterization of two antisense transcripts from the major immediate early region of rat cytomegalovirus.
We have identified and characterized two antisense transcripts from the rat cytomegalovirus (RCMV) major immediate early (MIE) region. These transcripts, designated IE-AS1 and IE-AS2, are complementary to part of the sense IE1 transcript. The IE-AS transcripts were first detected in peripheral blood leukocytes (PBL) of RCMV-infected rats at 7 days post-infection (pi) in the absence of IE1 transcription. Nevertheless, both the IE1 and IE-AS transcripts were found at the same time in the salivary glands of RCMV-infected rats at 7 and 120 days pi as well as in RCMV-infected rat embryo fibroblasts (REFs) at 48 h pi.


COVID-19 pandemic underscores role of green space in urban carbon dynamics.
For Detroit Michigan the arrival of COVID-19 led to intensive measures to prevent further spread of the virus resulting in consequent changes in traffic and energy use. We take advantage of these different emission scenarios to explore CO2 dynamics in a postindustrial city with a declining population and increasing green space. We present atmospheric CO2 concentration and net urban ecosystem exchange of CO2 (NUE) from a typical eddy covariance system and canopy greenness from a field camera on the Wayne State University campus in midtown Detroit. We categorized our study period (January 18, 2020-July 31, 2020) into three subperiods associated with the state-wide shelter-in-place order. Our results support that the city was a net carbon source throughout the period, particularly during the shelter-in-place period, although reduced traffic lowered CO2 concentrations and NUE. However, during the post-order period when traffic was highest, atmospheric CO2 concentrations and NUE were lowest, suggesting that the greening of urban vegetation may have greater carbon mitigation potential than lowering anthropogenic carbon emissions through traffic reductions.


A comparison of the chemical composition of mixed bacteria harvested from the liquid and solid fractions of rumen digesta.
Steers, equipped with simple rumen cannulas, were given diets of approximately equal parts of rolled barley and straw supplemented with urea. The diets provided sufficient estimated rumen degradable nitrogen (RDN; RDN:metabolizable energy values of 1:3) to maintain maximum microbial synthesis. In some experiments Na235SO4 was introduced into the rumen to label microbial protein. Rumen digesta samples were taken before feeding and mixed rumen bacteria were separated from the solid (solid-associated bacteria; SAB) and liquid (liquid-associated bacteria; LAB) fractions of digesta. The most effective method of removing SAB from the fibre was a combination of homogenizing and pummelling. This process did not affect the physical form or chemical composition of the bacteria. Samples of SAB contained significantly (P less than or equal to at least 0.05) less ash, total N, RNA and diaminopimelic acid (DAP) and significantly (P less than or equal to 0.01) more lipid than samples of LAB. Concentrations (g/kg dry matter) of ash, total N, RNA, DAP and lipid in SAB were approximately 87, 70, 35, 2.2 and 245 respectively. Corresponding values for LAB were 157, 80, 50, 3.8 and 124 respectively. RNA-N:total N and DAP-N:total N values in SAB were significantly lower than those in LAB (P less than or equal to 0.05 and 0.02 respectively). 35S:total N values were similar in both groups of bacteria. The importance of differences in constituent:total N values in the two groups of bacteria in relation to their use as indices of microbial protein synthesis is discussed.


The R215W mutation in NBS1 impairs gamma-H2AX binding and affects DNA repair: molecular bases for the severe phenotype of 657del5/R215W Nijmegen breakage syndrome patients.
Nijmegen breakage syndrome (NBS) is a genetic disorder characterized by chromosomal instability and hypersensitivity to ionising radiation. Compound heterozygous 657del5/R215W NBS patients display a clinical phenotype more severe than the majority of NBS patients homozygous for the 657del5 mutation. The NBS1 protein, mutated in NBS patients, contains a FHA/BRCT domain necessary for the DNA-double strand break (DSB) damage response. Recently, a second BRCT domain has been identified, however, its role is still unknown. Here, we demonstrate that the R215W mutation in NBS1 impairs histone gamma-H2AX binding after induction of DNA damage, leading to a delay in DNA-DSB rejoining. Molecular modelling reveals that the 215 residue of NBS1 is located between the two BRCT domains, affecting their relative orientation that appears critical for gamma-H2AX binding. Present data represent the first evidence for the role of NBS1 tandem BRCT domains in gamma-H2AX recognition, and could explain the severe phenotype observed in 657del5/R215W NBS patients.


ClassifyCNV: a tool for clinical annotation of copy-number variants.
Copy-number variants (CNVs) are an important part of human genetic variation. They can be benign or can play a role in human disease by creating dosage imbalances and disrupting genes and regulatory elements. Accurate identification and clinical annotation of CNVs is essential, however, manual evaluation of individual CNVs by clinicians is challenging on a large scale. Here, we present ClassifyCNV, an easy-to-use tool that implements the 2019 ACMG classification guidelines to assess CNV pathogenicity. ClassifyCNV uses genomic coordinates and CNV type as input and reports a clinical classification for each variant, a classification score breakdown, and a list of genes of potential importance for variant interpretation. We validate ClassifyCNV's performance using a set of known clinical CNVs and a set of manually evaluated variants. ClassifyCNV matches the pathogenicity category for 81% of manually evaluated variants with the significance of the remaining pathogenic and benign variants automatically determined as uncertain, requiring a further evaluation by a clinician. ClassifyCNV facilitates the implementation of the latest ACMG guidelines in high-throughput CNV analysis, is suitable for integration into NGS analysis pipelines, and can decrease time to diagnosis. The tool is available at https://github.com/Genotek/ClassifyCNV .


An x-ray microanalysis study of differences in concentration of elements in brain cells due to opiates, cell type, and subcellular location.
Adult female Sprague Dawley rats were divided into three groups and given two daily doses of: 1) morphine (25 mg/kg at each injection), or 2) methadone (5 mg/kg), or 3) saline for nine days. Two hours before the rats were killed on day 10, they were given a double dose of the drugs. Fifteen minutes before being killed, some of the morphine-treated rats were given the opiate antagonist naloxone (2.5 mg/kg), which caused a sudden arousal in these rats. At the time of killing, the preoptic-hypothalamic region was repidly removed and frozen in liquid propane to prevent translocation of elements in cells. Frozen 4-micrometer sections were cut, freeze-dried, and electron-probed in a scanning electron microscope, and energy-dispersive x-ray spectra were collected. The characteristic peak-to-continuum ratio for all detectable elements was determined in both the nucleus and the cytoplasm of ependymal cells and neurons. The data from nine cells of each type in each rat brain were then subjected to one- and three-way analysis of variance. The results show significant differences in the distribution of elements (sodium, magnesium, phosphorus, sulfur, chlorine, potassium, and calcium) which are dependent upon: 1) subcellular localization, 2) cell type, and particularly, 3) opiate treatment. The behavioral state produced by the opiates is correlated with the effects they have on intracellular concentration of several elements, most notably, sodium.


Juvenile hormone diol kinase, a calcium-binding protein with kinase activity, from the silkworm, Bombyx mori.
Juvenile hormone (JH) diol kinase (JHDK) is an important enzyme involved in the JH degradation pathway. Bombyx mori (Bommo)-JHDK cDNA (637bp) contains an open reading frame encoding a 183-amino acid protein, which reveals a high degree of identity to the two previously reported JHDKs. JHDK is similar to GTP-binding proteins with three conserved sequence elements involved in purine nucleotide binding, contains eight alpha-helices and three EF-hand motifs, and resembles the three-dimensional model of 2SCP and some other calcium-binding proteins. The Bommo-JHDK gene has only a single copy in the silkworm haploid genome, contains only one exon, and its 5'-upstream sequence does not have a JH response element. Although Bommo-JHDK is highly expressed in the gut of the silkworm, its mRNA expression remains at a constant level during larval development suggesting this enzyme is constitutive and not regulated by JH, at least at the transcriptional level. Recombinant Bommo-JHDK catalyzed the conversion of 10S-JH diol into JH diol phosphate, confirming its enzymatic function. Recombinant enzyme formed a dimer and had biochemical characteristics similar to other JHDKs. Bommo-JHDK, a calcium-binding protein with kinase activity, provides unique insights on how JH levels are regulated in the silkworm.


Ankle fracture: radiographic approach according to the Lauge-Hansen classification.
Ankle fractures account for 9 % of fractures (Clare in Foot Ankle Clin 13(4):593-610, 1) representing a significant portion of the trauma workload; proximal femoral fractures are the only lower limb fracture to present more frequently. Ankle fractures have a bimodal age distribution with peaks in younger males and older females (Arimoto and Forrester in AJR Am J Roentgenol 135(5):1057-1063, 2). There has been threefold increase in the incidence among elderly females over the past three decades (Haraguchi and Armiger in J Bone Joint Surg Am 91(4):821-829, 3). In 1950, Lauge-Hansen devised a classification of ankle fractures based on the position of the foot and the deforming force at the time of injury. This has been widely accepted by orthopedists, but is not in general use by radiologists. Identification of the fractures and classification of the type of injury allows diagnosis of the otherwise occult ligamentous injuries. Three radiographic views of the ankle (anteroposterior, mortise, and lateral) are necessary to classify an injury with the Lauge-Hansen system. Two additional criteria are also necessary: the position of the foot at the time of injury and the direction of the deforming force.


Classification of rice leaf blast severity using hyperspectral imaging.
Rice leaf blast is prevalent worldwide and a serious threat to rice yield and quality. Hyperspectral imaging is an emerging technology used in plant disease research. In this study, we calculated the standard deviation (STD) of the spectral reflectance of whole rice leaves and constructed support vector machine (SVM) and probabilistic neural network (PNN) models to classify the degree of rice leaf blast at different growth stages. Average accuracies at jointing, booting and heading stages under the full-spectrum-based SVM model were 88.89%, 85.26%, and 87.32%, respectively, versus 80%, 83.16%, and 83.41% under the PNN model. Average accuracies at jointing, booting and heading stages under the STD-based SVM model were 97.78%, 92.63%, and 92.20%, respectively, versus 88.89%, 91.58%, and 92.20% under the PNN model. The STD of the spectral reflectance of the whole leaf differed not only within samples with different disease grades, but also among those at the same disease level. Compared with raw spectral reflectance data, STDs performed better in assessing rice leaf blast severity.


Risk factors for recurrence after surgical repair of coarctation of the aorta in children: a single-center experience based on 51 children.
Coarctation of the aorta (CoA), is a congenital malformation, often combined with several cardiac abnormalities. At present, the operation effect is satisfactory, but postoperative restenosis is still a matter. Identification of risk factors for restenosis and prompt therapy adjustments may improve patient outcomes. A retrospective clinical study of patients under 12 who had CoA repair in 2012-2021, with a randomized cohort population of 475 patients. A total of 51 patients (M/F: 30/21) with a mean age of 5.33 (2.00-15.00) months and a median weight of 5.60 (4.20-10.00) kg. The mean follow-up was 8.93 (3.77-19.37) months. Patients were divided into 2 groups: no-restenosis (n-reCoA) (G1, 38 patients) and restenosis (reCoA) (G2, 13 patients). ReCoA was defined as a restenosis requiring interventional or surgery or a pressure gradient >20 mmHg at the repair site as reported by B-ultrasound with the presence of an upper and lower limb blood pressure gradient or growing dysplasia. The overall reCoA incidence was 25% (13/51). In multivariate COX regression, smaller preoperative z-score of the ascending aorta (P = 0.009, HR = 0.68) and transverse aortic arch (P = 0.015, HR = 0.66), arm-leg systolic pressure gradient ≥12.5 mmHg at discharge (P = 0.003, HR = 1.09) were independent risk factors for reCoA. The overall outcome of CoA surgery is successful. Smaller preoperative z-score of the ascending aorta and transverse aortic arch, and an arm-leg systolic pressure gradient ≥12.5 mmHg at discharge increase reCoA risk, and closer follow-up for such patients are required especially within 1 postoperative year.


Labour outcome of low-risk multiparas of 40 years and older. A case-control study.
Between January 1, 1994 and December 31, 1997 a total of 76 low-risk multiparas of 40 years and older with spontaneous onset of labour were delivered and were compared with 152 younger (25-30 years-old) low-risk multiparas of similar parity in a case-control study. The labour and perinatal outcomes of the 2 groups were compared. The duration of the first stage of labour was longer (233 minutes versus 149 minutes, p<0.0005) in the older women. Significantly more labours of older multiparas were complicated by intrapartum fetal distress (6.6% versus 1.3%, p<0.05); received intramuscular analgesia (11.8% versus 2.6%, p<0.01); and had operative deliveries (17.1% versus 4.6%, p<0.01). The incidence of instrumental delivery (11.8% versus 3.9%, p<0.05) and Caesarean section (5.3% versus 0.7%, p<0.05) were higher among older multiparas. The incidences of Syntocinon augmentation of labour, prolonged second stage, episiotomy and third stage complications such as perineal tear, primary postpartum haemorrhage, and retained placenta were similar in both groups. Both groups had similar perinatal outcomes. These women should be treated as others with high-risk pregnancies with appropriate careful attention during labour such as continuous fetal heart rate monitoring. They should be counselled to be made aware of the increased risks during labour.


In vitro production of a hybrid monoclonal antibody that preferentially binds to cells that express both HLA-A2 and HLA-B7.
A hybrid mouse monoclonal IgGl having one low affinity combining site for HLA-A2 and one low affinity combining site for HLA-B7 was made by the chemical method of Nisonoff and Palmer (Science 143:376,1964). This involved selective reduction of interchain disulphides, a splitting of the IgGl into half molecules at low pH and ionic strength, and reassociation of the half molecules by neutralization. Serologically active hybrids were separated from parental IgGl by an absorbtion procedure and recovered in about 10% yield. The hybrid discriminated between cells that express either HLA-A2 or HLA-B7 from cells that express both A2 and B7. This is because it could bind bivalently to the cell with both A2 and B7 but could only bind with a single combining site to cells expressing A2 or B7. The consequence of these different modes of attachment was to give up to sevenfold greater binding to the cell expressing A2 and B7 in comparison to the cell expressing only A2 or B7.


Radiation-induced solitary rectal ulcer.
A case of radiation-induced solitary rectal ulcer is presented. The unusual posterior location with involvement of the internal iliac artery produced massive lower intestinal hemorrhage. Temporary control was obtained with percutaneous arterial catheter embolization, but definitive control necessitated surgical resection.


Mini-open carpal tunnel release: technique, feasibility and clinical outcome compared to the conventional procedure in a long-term follow-up.
We sought to evaluate the findings of our anatomically landmarks based mini-open procedure (MCTR) through a palmar approach and to compare its outcome and practicability to the conventional method (OCTR). The study consisted of 100 matched patients (n = 50 MCTR, n = 50 OCTR) with a minimum follow-up of three years. The outcome was characterized via the Disabilities of Arm, Shoulder and Hand Score (DASH), Symptom Severity Scale (SSS), Functional Status Scale (FSC), and Visual Analogue Scale (VAS). All adverse events were observed. An alpha of 0.05 and a confidence level of 95% were set for statistical analyses. Both techniques showed comparable functional results in a long-term period (mean follow-up MCTR: 60 months and OCTR: 54 months). MCTR versus OCTR at mean: DASH: 4.6/8.3 (p = 0.398), SSS: 1.3/1.2 (p = 0.534), FSC: 1.3/1.2 (p = 0.617), VAS: 0.4/0.7 (p = 0.246). The MCTR convinced through a lower rate of scar sensibility (MCTR: 0% vs. OCTR: 12%, 0/50 vs. 6/50; p = 0.007) and pillar pain, as well as a shortened recovery period and surgical time relative to the OCTR. Low complication rates were observed in both groups, no recurrences had to be documented. The MCTR procedure revealed a similar good clinical outcome as the conventional technique. MCTR is a minimally-invasive, reliable, fast and simple procedure with an obvious benefit regarding scar sensibility.


Coronary artery wall imaging in mice using osmium tetroxide and micro-computed tomography (micro-CT).
The high spatial resolution of micro-computed tomography (micro-CT) is ideal for 3D imaging of coronary arteries in intact mouse heart specimens. Previously, micro-CT of mouse heart specimens utilized intravascular contrast agents that hardened within the vessel lumen and allowed a vascular cast to be made. However, for mouse coronary artery disease models, it is highly desirable to image coronary artery walls and highlight plaques. For this purpose, we describe an ex vivo contrast-enhanced micro-CT imaging technique based on tissue staining with osmium tetroxide (OsO(4) ) solution. As a tissue-staining contrast agent, OsO(4) is retained in the vessel wall and surrounding tissue during the fixation process and cleared from the vessel lumens. Its high X-ray attenuation makes the artery wall visible in CT. Additionally, since OsO(4) preferentially binds to lipids, it highlights lipid deposition in the artery wall. We performed micro-CT of heart specimens of 5- to 25-week-old C57BL/6 wild-type mice and 5- to 13-week-old apolipoprotein E knockout (apoE(-/-) ) mice at 10 μm resolution. The results show that walls of coronary arteries as small as 45 μm in diameter are visible using a table-top micro-CT scanner. Similar image clarity was achieved with 1/2000th the scan time using a synchrotron CT scanner. In 13-week-old apoE mice, lipid-rich plaques are visible in the aorta. Our study shows that the combination of OsO(4) and micro-CT permits the visualization of the coronary artery wall in intact mouse hearts.


Prevalence of HLA DQB1*0602 allele in patients with migraine.
Studies have shown a high prevalence of migraine among narcoleptic patients. HLA-DQB1*0602 and HLA DRB1 alleles are closely associated with narcolepsy. An increase in the HLA-DRB1 allele frequency in patients with visual aura has raised greater awareness of the genetic background in migraine. Since the regions DR and DQ of the HLA are in tightly linkage desiquilibrium we hypothesize that HLA-DQB1*0602 might be associated to the pathophysiology of migraine. We analyzed the presence of HLA DQB1*0602 allele in 50 healthy subjects with no history of migraine, 53 patients with migraine without aura and 52 patients with migraine with aura. There was no difference in the frequency of HLA DQB1*0602 allele when control subjects and all patients were compared. We failed to note any difference in frequencies when comparing migraine patients with and without aura. Further studies with different patient populations, with other hypothalamic markers (melatonin, hypocretin) in migraine patients may shed light on to its pathophysiology.


A new mutation in the GNAL gene in familial dystonia presenting with mental symptoms.
GNAL mutations (DYT25) have lately been identified as the firstly proven cause of focal adult-onset dystonia. We report here a new mutation in the GNAL gene in two siblings with dystonia. The new mutation is called NM 001,142,339:c.97C > T. Our research emphasizes the possible effects of new mutation on disease risk and the significance of genetic tests for GNAL mutations in confirming the molecular diagnosis.


Oxidative damage in human gingival fibroblasts exposed to cigarette smoke.
Cigarette smoke, a complex mixture of over 7000 chemicals, contains many components capable of eliciting oxidative stress, which may induce smoking-related disorders, including oral cavity diseases. In this study, we investigated the effects of whole (mainstream) cigarette smoke on human gingival fibroblasts (HGFs). Cells were exposed to various puffs (0.5-12) of whole cigarette smoke and oxidative stress was assessed by 2',7'-dichlorofluorescein fluorescence. The extent of protein carbonylation was determined by use of 2,4-dinitrophenylhydrazine with both immunocytochemical and Western immunoblotting assays. Cigarette smoke-induced protein carbonylation exhibited a puff-dependent increase. The main carbonylated proteins were identified by means of two-dimensional electrophoresis and MALDI-TOF mass spectrometry (redox proteomics). We demonstrated that exposure of HGFs to cigarette smoke decreased cellular protein thiols and rapidly depleted intracellular glutathione (GSH), with a minimal increase in the intracellular levels of glutathione disulfide and S-glutathionylated proteins, as well as total glutathione levels. Mass spectrometric analyses showed that total GSH consumption is due to the export by the cells of GSH-acrolein and GSH-crotonaldehyde adducts. GSH depletion could be a mechanism for cigarette smoke-induced cytotoxicity and could be correlated with the reduced reparative and regenerative activity of gingival and periodontal tissues previously reported in smokers.


IL-13 induces the expression of the alternative activation marker Ym1 in a subset of testicular macrophages.
Macrophages are thought to play an important role in the maintenance of immune privilege in the testis, which functions to prevent immune responses to developing sperm. Two populations of macrophages are known to exist in the testis, one of which exhibits immunosuppressive activity. Macrophages that are alternatively activated with either IL-4 or IL-13 have been shown to be anti-inflammatory and promote wound healing. Expression of the Ym1 protein is an established marker of alternatively activated macrophages. Testicular macrophages were examined for expression of Ym1 protein, and it was found to be highly expressed in a subpopulation of CD11b(+) cells. Furthermore, we have shown that Ym1 protein expression in the testis is dependent upon IL-13R signaling, and that IL-13 is produced in the testis. These data suggest that IL-13 plays a role in testicular immune privilege by the maintenance of an alternatively activated macrophage population.


Differences in the respiratory response to temperature and hypoxia across four life-stages of the intertidal porcelain crab Petrolisthes laevigatus.
For aquatic breathers, hypoxia and warming can act synergistically causing a mismatch between oxygen supply (reduced by hypoxia) and oxygen demand (increased by warming). The vulnerability of these species to such interactive effects may differ during ontogeny due to differing gas exchange systems. This study examines respiratory responses to temperature and hypoxia across four life-stages of the intertidal porcelain crab Petrolisthes laevigatus. Eggs, megalopae, juveniles and adults were exposed to combinations of temperatures from 6 to 18 °C and oxygen tensions from 2 to 21 kPa. Metabolic rates differed strongly across life-stages which could be partly attributed to differences in body mass. However, eggs exhibited significantly lower metabolic rates than predicted for their body mass. For the other three stages, metabolic rates scaled with a mass exponent of 0.89. Mass scaling exponents were similar across all temperatures, but were significantly influenced by oxygen tension (the highest at 9 and 14 kPa, and the lowest at 2 kPa). Respiratory responses across gradients of oxygen tension were used to calculate the response to hypoxia, whereby eggs, megalopae and juveniles responded as oxyconformers and adults as oxyregulators. The thermal sensitivity of the metabolic rates (Q10) were dependent on the oxygen tension in megalopae, and also on the interaction between oxygen tension and temperature intervals in adults. Our results thus provide evidence on how the oxygen tension can modulate the mass dependence of metabolic rates and demonstrate changes in respiratory control from eggs to adults. In light of our results indicating that adults show a good capacity for maintaining metabolism independent of oxygen tension, our study highlights the importance of assessing responses to multiple stressors across different life-stages to determine how vulnerability to warming and hypoxia changes during development.


Evaluation of the Manchester classification system for chronic pancreatitis.
Classifications of chronic pancreatitis based on either histologic (Marseilles) or endoscopic (Cambridge) criteria are not widely used. The present study describes the development and validation of a three-stage clinical categorical classification system for chronic pancreatitis. Patients with a diagnosis of chronic pancreatitis (577.1: ICD-9) for 1993 were identified from records of the Hepatopancreaticobiliary service at a University hospital. Endoscopic or CT evidence of chronic pancreatitis were mandatory for inclusion. Forty one patients met the criteria and were categorized according to a 3-stage system as mild, moderate or end-stage disease. The clinical course over the subsequent decade was followed by chart review with re-categorization of stage at each review. At the outset of the study, 18 (44%) patients were categorised as having mild disease, 19 (46%) as moderate and 4 (10%) as end-stage. The number of patients with mild disease fell over the subsequent 5 years and at the end of the 10-year chart study period, no patients were categorised as mild. The number of patients with diabetes at the outset of the study period was 2 (5%). At two years this was 3 (7%), five years 10 (24%) and 10 years was 25 (61%). These results show that the Manchester classification of chronic pancreatitis is both practical and feasible and now requires prospective evaluation and independent appraisal by other centres.


Unnecessary wait hours: a novel ED system-level metric.
Emergency department (ED) operations leaders are under increasing pressure to make care delivery more efficient. Publicly reported ED efficiency metrics are traditionally patient centred and do not show situational or facility-based improvement opportunities. We propose the consideration of a novel metric, the 'Number of Unnecessary Waits (NUW)' and the corresponding 'Unnecessary Wait Hours (UWH)', to measure space efficiency, and we describe how we used NUW to evaluate operational changes in our ED. UWH summarises the relationship between the number of available rooms and the number of patients waiting by returning a value equal to the number of unnecessary patient waits. We used this metric to evaluate reassigning a clinical technician assistant (CTA) to the new role of flow CTA. We retrospectively analysed 3.5 months of data from before and after creation of the flow CTA. NUW metric analysis suggested that the flow CTA decreased the amount of unnecessary wait hours, while higher patient volumes had the opposite effect. Situational system-level metrics may provide a new dimension to evaluating ED operational efficiencies. Studies focussed on system-level metrics to evaluate an ED practice are needed to understand the role these metrics play in evaluation of a department's operations.


Direct reporting of laboratory test results to patients by mail to enhance patient safety.
Missed test results are common in clinical practice and compromise patient safety. Direct reporting, whereby testing centers systematically notify both patients and providers of important test results, constitutes a potential solution, but provider acceptance is unknown. To assess provider interest in direct reporting of selected test results and how interest varied across different tests. Survey of primary care physicians at a tertiary care academic medical center. Five-point Likert scores were used to gauge each physician's interest (1 = not at all interested to 5 = very interested) in scenarios pertaining to the direct reporting of 3 diagnostic tests of low (DXA scan), intermediate (genital herpes testing), and high (breast biopsy) "emotional impact" and whether interest varied with each test's result (normal vs abnormal). Physicians were also asked to cite specific advantages and disadvantages of direct reporting. The response rate was 73% (148/202). Physician interest in direct reporting decreased progressively as scenarios shifted from low (DXA scan) to high (breast biopsy) emotional impact (P < .001); interest in direct reporting was also higher when results were normal rather than abnormal (P < .001). Common advantages of direct reporting cited by respondents were reductions in workload (selected by 75% of respondents) and reductions in missed diagnoses (38%). The most common concerns were that patients would become unnecessarily frightened (70%) and would seek unreliable information (65%). Direct reporting of selected test results to patients is one system for insuring that important results are not missed, but implementation should consider the specific test in question, the test result, and provider preferences.


Automated apparent diffusion coefficient analysis for genotype prediction in lower grade glioma: association with the T2-FLAIR mismatch sign.
The prognosis of lower grade glioma (LGG) patients depends (in large part) on both isocitrate dehydrogenase (IDH) gene mutation and chromosome 1p/19q codeletion status. IDH-mutant LGG without 1p/19q codeletion (IDHmut-Noncodel) often exhibit a unique imaging appearance that includes high apparent diffusion coefficient (ADC) values not observed in other subtypes. The purpose of this study was to develop an ADC analysis-based approach that can automatically identify IDHmut-Noncodel LGG. Whole-tumor ADC metrics, including fractional tumor volume with ADC > 1.5 × 10-3mm2/s (VADC>1.5), were used to identify IDHmut-Noncodel LGG in a cohort of N = 134 patients. Optimal threshold values determined in this dataset were then validated using an external dataset containing N = 93 cases collected from The Cancer Imaging Archive. Classifications were also compared with radiologist-identified T2-FLAIR mismatch sign and evaluated concurrently to identify added value from a combined approach. VADC>1.5 classified IDHmut-Noncodel LGG in the internal cohort with an area under the curve (AUC) of 0.80. An optimal threshold value of 0.35 led to sensitivity/specificity = 0.57/0.93. Classification performance was similar in the validation cohort, with VADC>1.5 ≥ 0.35 achieving sensitivity/specificity = 0.57/0.91 (AUC = 0.81). Across both groups, 37 cases exhibited positive T2-FLAIR mismatch sign-all of which were IDHmut-Noncodel. Of these, 32/37 (86%) also exhibited VADC>1.5 ≥ 0.35, as did 23 additional IDHmut-Noncodel cases which were negative for T2-FLAIR mismatch sign. Tumor subregions with high ADC were a robust indicator of IDHmut-Noncodel LGG, with VADC>1.5 achieving > 90% classification specificity in both internal and validation cohorts. VADC>1.5 exhibited strong concordance with the T2-FLAIR mismatch sign and the combination of both parameters improved sensitivity in detecting IDHmut-Noncodel LGG.


Infection by exosome-carried Coxsackievirus B3 induces immune escape resulting in an aggravated pathogenesis.
Increasing evidence has shown that extracellular vesicles or exosomes released from virus-infected cells contain viral particles, genomes, or other pathogenic factors that move to neighbor cells, contributing to virus dissemination and productive infection. Our recent study demonstrated that exosomes carrying CVB3 virions exhibited greater infection efficiency than free virions because they accessed various entry routes, overcoming restrictions to viral tropism. However, the pathogenicity of exosomes carried CVB3 and their effect on immunological properties have not yet been completely explained. In the current study, we sought to explore whether exosomes exert their effect on the CVB3-induced pathogenesis or evade the immune attack. Our results showed that exosomes-carried CVB3 could effectively infect viral receptor-negative immune cells in vivo, resulting in inducing immune system loss. Importantly, the exosomes-carried CVB3 had the ability to escape the neutralizing antibodies activity resulting in inducing the severe onset of myocarditis. Using the genetically engineered mouse with deficiency of exosomes, we observed that the exosomes-carried CVB3 reinforced an aggravated pathogenesis. By understanding how exosomes promote the course of viral disease, clinical applications of exosomes can be developed.


Quality of life in patients with multifocal motor neuropathy from Serbia.
Multifocal motor neuropathy (MMN) is a rare, chronic disorder with potentially severe and progressive disability, which may affect patients' quality of life (QoL). Since there is still small number of studies that predominantly investigated QoL in patients with MMN, we sought to analyze QoL in these patients. Our study comprised 17 patients diagnosed with MMN at the same clinic. Following scales were used: SF-36 questionnaire, INCAT disability scale, Krupp's Fatigue Severity scale, and Beck Depression Inventory. Physical domains of QoL were slightly more affected than mental ones, but with no statistical significance (64.8 ± 22.3 vs. 70.0 ± 19.5, p > 0.05). Total SF-36 score was 69.2 ± 19.9. INCAT arm disability score at testing was found to correlate with the total SF-36 score (rho = -0.603, p < 0.05). INCAT arm disability score at diagnosis (rho = -0.57, p < 0.05) and at testing (rho = -0.48, p = 0.05) correlated with physical composite score (PCS). Disease duration (rho = -0.51, p < 0.05) and INCAT arm disability score at testing (rho = -0.60, p = 0.01) were associated with mental composite score (MCS). QoL in patients with MMN was reduced, especially in physical domains. Although arm disability was the most significant parameter which affected QoL of MMN patients in both physical and mental aspects, longer disease duration should not be underestimated as a psychological burden for these patients.


Prenatal diagnosis of GM2-gangliosidosis B1 variant.
Prenatal diagnosis in a pregnancy at risk for a juvenile B1 variant of GM2-gangliosidosis was carried out. The biochemical study of the cultured amniocytes and the affected fetal brain is reported. The results obtained show that the sulphated artificial substrate can be used in the diagnosis of B1 variant, but not the neutral one. The accumulation of GM2-ganglioside in the fetal brain of the B1 juvenile form and an infantile form of GM2-gangliosidosis (0 variant) was compared.


Comparison of emergency department time performance between a Canadian and an Australian academic tertiary hospital.
To compare performance and factors predicting failure to reach Ontario and Australian government time targets between a Canadian (Sunnybrook Hospital) and an Australian (Austin Health) academic tertiary-level hospitals in 2012, and to assess for change of factors and performance in 2016 between the same hospitals. This was a retrospective, observational study of patient administrative data in two calendar years. The main outcome measure was reaching Ontario and Australian ED time targets for admissions, high and low urgency discharges. Secondary outcomes were factors predicting failure to reach these targets. Between 2012 and 2016, Sunnybrook and Austin experienced increased patient volume of 10.2% and 19.2%, respectively. Bed capacity decreased at Sunnybrook (-10.8%) but increased at the Austin (+30.3%). For both years, Austin failed to achieve the Australian time target, but succeeded for all Ontario targets except for low urgency discharges. Sunnybrook failed all targets irrespective of year. The top factors for failing Ontario ED length-of-stay targets for both hospitals in 2012 and 2016 were bed request greater than 6 h, access block greater than 1 h, use of cross-sectional imaging, consultation and waiting for the emergency physician greater than 2 h. Austin outperformed Sunnybrook for Ontario and Australian government time targets. Both hospitals failed the Australian targets. Factors predicting failure to achieve targets were different between hospitals, but were mainly clinical resources. Sunnybrook focussed on increasing human resources. Austin focussed on increasing human resources, observation unit and hospital beds. Intrinsic hospital characteristics and infrastructure influenced target success.


Surgical Technique: Supraclavicular First Rib Resection.
Thoracic outlet syndrome is a condition of compression involving the brachial plexus and subclavian vessels. Although there are multiple surgical approaches to address thoracic outlet decompression, supraclavicular first rib resection with scalenectomy and brachial plexus neurolysis allow for complete exposure of the first rib, brachial plexus, and vasculature. This technique is described in detail. This approach is safe and can produce excellent outcomes in all variants of thoracic outlet syndrome.


Microcystin-leucine-arginine promotes the development of gallbladder carcinoma via regulating ELAC2.
Gallbladder carcinoma (GBC) is the most prevalent cancer of the bile tract, with unexpected GBC accounting for almost half of all GBC cases in some tertiary medical centers. Although the involvement of microcystin-leucine-arginine (MC-LR) in the development of intrahepatic cholangiocarcinoma has been established, there is a paucity of data regarding its association with GBC. The present study aims to investigate whether MC-LR level in the gallbladder of patients is associated with GBC development and, if so, to characterize the underlying mechanism in GBC cells. Our clinical data revealed that MC-LR level was significantly increased in GBC patients compared to patients with gallbladder stones only (P = 0.009). Moreover, our findings demonstrated that MC-LR could promote the proliferation and metastasis of human GBC cell lines. Furthermore, ELAC2 was identified as a critical mRNA involved in GBC progression through RNA sequencing. Collectively, our study suggests that MC-LR might be involved in the development of GBC by modulating the expression of ELAC2.