PREVALENCE AND RISK FACTORS OF ELLIPSOID ZONE DAMAGE AFTER PARS PLANA VITRECTOMY FOR IDIOPATHIC EPIRETINAL MEMBRANE.
To assess factors associated with external limiting membrane (ELM)/ellipsoid zone (EZ) damage after pars plana vitrectomy (PPV) for epiretinal membrane and evaluate ELM/EZ damage impact on functional and anatomical outcomes. Patients who underwent PPV with epiretinal membrane ± inner limiting membrane peeling were retrospectively analyzed. Best-corrected visual acuity and central macular thickness were longitudinally collected based on the available follow-up. Demographic data, clinical data, and surgical details were included in logistic regression models having ELM/EZ damage as a binary outcome. Overall, 179 eyes (171 patients) were included; 93 had a 12-month follow-up. Thirty-four eyes (19%) had ELM/EZ damage after surgery; in nine eyes (5%), ELM/EZ damage persisted at 12 months. Phacoemulsification during PPV (odds ratio = 6.97; P = 0.007) and ELM/EZ damage before PPV (odds ratio = 6.91; P = 0.007) were risk factors for postoperative ELM/EZ disruption. Thicker outer nuclear layer (P = 0.002), thicker ectopic inner foveal layer (P < 0.001), and higher endoillumination power (P = 0.03) were associated with slower visual recovery. Inner limiting membrane peeling (P = 0.04) was associated with slower anatomical recovery. Cataract extraction and outer retinal damage before PPV are associated with higher risk of photoreceptor damage postoperatively. The rate of visual improvement varies as a function of retinal layers' distortion before surgery; additional retinal manipulation slows restoration of the macular anatomy.


Exosomal microRNA-222-3p increases UVB sensitivity of lens epithelium cells by suppressing MGMT.
Age-related cataract (ARC) is a leading cause of blindness worldwide with multiple pathogenic factors. Oxidative damage of lens epithelium cells (LECs) is one of the well-accepted pathogenesis of ARC which can be regulated by DNA repair genes (DRGs). The present research aimed to clarify the regulatory mechanism of exosomal microRNAs (miRNAs) on DRGs in LECs. The LECs oxidative damage model was established by UVB-irradiation on SRA01/04 (human lens epithelium cell line). Exosomes from UVB-irradiated cells (UVB-exo) and exosomes from normal control cells (NC-exo) were collected from the culture medium. To explore the functions of LECs exosomes, SRA01/04 were incubated with UVB-exo/NC-exo. Then, we detected SRA01/04 proliferation, viability and apoptosis respectively using 5'-ethynyl-2'-deoxyuridine (EdU), cell-counting kit-8 (CCK-8) and TdT-mediated dUTP Nick-End Labeling (TUNEL) assay. Next, the miRNA expression profiles of UVB-exo and NC-exo were identified by miRNA microarrays. RNA expression in exosomes, cells, and clinical samples was verified by qRT-PCR. The location and expression of MGMT and CD63 proteins were detected by immunofluorescence and western blot. The 3'UTR regulation of miR-222-3p to MGMT was verified by luciferase analyses. MGMT down-regulated while miR-222-3p up-regulated in LECs sub-central anterior capsule from ARC lenses. MGMT and miR-222-3p expressions in central and peripheral LECs from anterior lens capsules were differential. UVB-exo can transport the up-regulated miR-222-3p from oxidative-damaged LECs to normal LECs, which could suppress MGMT expression and increase UVB sensitivity of LECs. Findings on exosomal miRNA functions provided novel insights into pathogenesis of ARC. Exosomal miR-222-3p can be a potential target for prevention and cure of ARC.


MDFF-Net: A multi-dimensional feature fusion network for breast histopathology image classification.
Breast cancer is a common malignancy and early detection and treatment of it is crucial. Computer-aided diagnosis (CAD) based on deep learning has significantly advanced medical diagnostics, enhancing accuracy and efficiency in recent years. Despite the convenience, this technology also has certain limitations. When the morphological characteristics of the patient's pathological section are not evident or complex, certain small lesions or cells deep within the lesion cannot be recognized, and misdiagnosis is prone to occur. As a result, MDFF-Net, a CNN-based multidimensional feature fusion network, is proposed. The model consists of a one-dimensional feature extraction network, a two-dimensional feature extraction network, and a feature fusion classification network. The basic part of the two-dimensional feature extraction network is stacked by modules integrated with multi-scale channel shuffling networks and channel attention modules. Furthermore, inspired by natural language processing, this model integrates a one-dimensional feature extraction network to extract detailed information in the image to avoid misdiagnosis caused by insufficient information extraction such as cell morphological characteristics and differentiation degree. Finally, the extracted one-dimensional and two-dimensional features are fused in the feature fusion network and employed for the final classification. The effectiveness of MDFF-Net and classical classification models were evaluated on the BreakHis and the BACH datasets. According to experimental results, MDFF-Net achieves an accuracy of 98.86% on the BreakHis and 86.25% on the BACH dataset. Furthermore, to further assess the effectiveness of the model in other classification tasks, the colon cancer and the lung cancer datasets were employed for additional experiments, achieving a classification accuracy of 100% in both cases.


Immobilization of chromium ore processing residue by alkali-activated composite binders and leaching characteristics.
Chromium ore processing residue (COPR) is classified as hazardous solid waste because of the leachable Cr(VI). Cementitious materials are often used to solidify and stabilize heavy metals. However, most of them focus on the leaching concentration of particles after solidification and stabilization and lack research on leaching characteristics. This study investigated the leaching characteristics of heavy metals in three simulated environments (HJ557-2010, HJ/T299-2007, TCLP) after immobilizing COPR with composite binders. Industrial solid waste coal fly ash and lead-zinc smelting slag are used to prepare composite binders through alkali activation technology. Compressive strength, particle leaching toxicity, acid neutralization capability, and semi-dynamic leaching test are used to evaluate the performance of the solidified body. The solidified body can be applied to building materials or treated as general industrial waste. Heavy metals are mainly released from the matrix by surface washing at a low rate. The analysis results, including XRD, FTIR, and SEM-EDS, show that chemical binding and physical encapsulation are the main immobilizing mechanisms to realize the coordinated disposal of Zn and Cr(VI).


Fragment Linker Prediction Using the Deep Encoder-Decoder Network for PROTACs Drug Design.
A drug discovery and development pipeline is a prolonged and complex process that remains challenging for both computational methods and medicinal chemists and has not been able to be resolved using computational methods. Deep learning has been utilized in various fields and achieved tremendous success in designing novel molecules in the pharmaceutical industry. Herein, we use state-of-the-art techniques to propose a deep neural network, AIMLinker, to rapidly design and generate meaningful drug-like proteolysis targeting chimeras (PROTACs) analogs. The model extracts the structural information from the input fragments and generates linkers to incorporate them. We integrate filters in the model to exclude nondruggable structures guided via protein-protein complexes while retaining molecules with potent chemical properties. The novel PROTACs subsequently pass through molecular docking, taking root-mean-square deviation (RMSD), relative Gibbs free energy (ΔΔGbinding), molecular dynamics (MD) simulation, and free energy perturbation (FEP) calculations as the measurement criteria for testing the robustness and feasibility of the model. The generated novel PROTACs molecules possess similar structural information with superior binding affinity to the binding pockets compared to the existing CRBN-dBET6-BRD4 ternary complexes. We demonstrate the effectiveness of the methodology of leveraging AIMLinker to design novel compounds for PROTACs molecules exhibiting better chemical properties compared to the dBET6 crystal pose.


Augmented reality with HoloLens in parotid surgery: how to assess and to improve accuracy.
Augmented reality improves planning and execution of surgical procedures. The aim of this study was to evaluate the feasibility of a 3D augmented reality hologram in live parotic surgery. Another goal was to develop an accuracy measuring instrument and to determine the accuracy of the system. We created a software to build and manually align 2D and 3D augmented reality models generated from MRI data onto the patient during surgery using the HoloLens® 1 (Microsoft Corporation, Redmond, USA). To assess the accuracy of the system, we developed a specific measuring tool applying a standard electromagnetic navigation device (Fiagon GmbH, Hennigsdorf, Germany). The accuracy of our system was measured during real surgical procedures. Training of the experimenters and the use of fiducial markers significantly reduced the accuracy of holographic system (p = 0.0166 and p = 0.0132). Precision of the developed measuring system was very high with a mean error of the basic system of 1.3 mm. Feedback evaluation demonstrated 86% of participants agreed or strongly agreed that the HoloLens will play a role in surgical education. Furthermore, 80% of participants agreed or strongly agreed that the HoloLens is feasible to be introduced in clinical routine and will play a role within surgery in the future. The use of fiducial markers and repeated training reduces the positional error between the hologram and the real structures. The developed measuring device under the use of the Fiagon navigation system is suitable to measure accuracies of holographic augmented reality images of the HoloLens.


Attractor-state itinerancy in neural circuits with synaptic depression.
Neural populations with strong excitatory recurrent connections can support bistable states in their mean firing rates. Multiple fixed points in a network of such bistable units can be used to model memory retrieval and pattern separation. The stability of fixed points may change on a slower timescale than that of the dynamics due to short-term synaptic depression, leading to transitions between quasi-stable point attractor states in a sequence that depends on the history of stimuli. To better understand these behaviors, we study a minimal model, which characterizes multiple fixed points and transitions between them in response to stimuli with diverse time- and amplitude-dependencies. The interplay between the fast dynamics of firing rate and synaptic responses and the slower timescale of synaptic depression makes the neural activity sensitive to the amplitude and duration of square-pulse stimuli in a nontrivial, history-dependent manner. Weak cross-couplings further deform the basins of attraction for different fixed points into intricate shapes. We find that while short-term synaptic depression can reduce the total number of stable fixed points in a network, it tends to strongly increase the number of fixed points visited upon repetitions of fixed stimuli. Our analysis provides a natural explanation for the system's rich responses to stimuli of different durations and amplitudes while demonstrating the encoding capability of bistable neural populations for dynamical features of incoming stimuli.


Congenitally corrected transposition with left ventricular outflow obstruction and cardiac malposition: One-and-a-half ventricular repair vs. Fontan pathway?
This study was to assess the mid-term results of the one-and-a-half ventricular repair (hemi-Mustard and bidirectional Glenn procedures combined with the Rastelli procedure) and Fontan pathway for correcting congenitally corrected transposition of great artery (ccTGA) patients with left ventricular outflow tract obstruction (LVOTO) and cardiac malposition. In this retrospective study, 74 consecutive ccTGA with LVOTO and cardiac malposition underwent the one-and-a-half ventricular repair (group A; 33 cases) and Fontan operation (group B; 41 cases) between October 2011 and March 2018. The Median follow-up time was 49 (20-84) and 42 (7-85) months in groups A and B, respectively. To estimate excise tolerance the 6-min walk test (MWT) was performed. No in-hospital death. Compared with group A, group B have significantly less CPB, mechanical ventilation time, and intensive care unit stay, but prolonged pleural effusions developed more frequently in Group B. The survival probability was 90.2% (95% CI, 80.2-100%) and 97.2% (95% CI, 92-100%) at 7 years (p = 0.300) in group A and B. The probability of freedom from re-intervention were 80.6% (95% CI, 66.5-97.6%) and 97.2% (95% CI, 92-100%) at 7 years (p = 0.110). Longitudinal repeated measured echo data at every follow-up time shows that group A has more systemic ventricular EF% (p < 0.001) and less moderate systemic ventricular valve regurgitation (p < 0.001) compared with group B. Estimated by 6 MWT, group A has better outcomes for 6-min walk distance. For correction of ccTGA with LVOTO and cardiac malposition, the one-and-a-half ventricular repair had superior midterm heart function and excise tolerance.


Effect of Desmodium adscendens fraction 3 on contractions of respiratory smooth muscle.
The third flash chromatography fraction prepared from an aqueous extract of Desmodium adscendens leaves (DAF3) was evaluated for pharmacological activity using contractions of tracheal spirals and lung parenchymal strips caused by ovalbumin, arachidonic acid, histamine and carbachol. DAF3 inhibited both the early and late phases of antigen-induced contractions of both tissues dose-dependently, but had no effect on arachidonic-induced contractions. With tracheal spirals, the inhibition occurred whether the tissue was pretreated with indomethacin or not. On tracheal spirals, the maximum contraction (Emax) caused by histamine and carbachol were enhanced, but the pD2 value for histamine was not affected while that for carbachol was decreased. DAF3 had no effect on Emax or pD2 values for these agonists on lung parenchymal strips. The results suggest that DAF3 may inhibit the release of free arachidonic acid.


Comparative Mapping of GABA-Immunoreactive Neurons in the Buccal Ganglia of Nudipleura Molluscs.
Phylogenetic comparisons of neurotransmitter distribution are important for understanding the ground plan organization of nervous systems. This study describes the γ-aminobutyric acid (GABA)-immunoreactive (GABA-ir) neurons in the buccal ganglia of six sea slug species (Mollusca, Gastropoda, Euthyneura, Nudipleura). In the nudibranch species, Hermissenda crassicornis, Tritonia diomedea, Tochuina tetraquetra, and Dendronotus iris, the number of GABA-ir neurons was highly consistent. Another nudibranch, Melibe leonina, however, contained approximately half the number of GABA-ir neurons. This may relate to its loss of a radula and its unique feeding behavior. The GABA immunoreactivity in a sister group to the nudibranchs, Pleurobranchaea californica, differed drastically from that of the nudibranchs. Not only did it have significantly more GABA-ir neurons but it also had a unique GABA distribution pattern. Furthermore, unlike the nudibranchs, the Pleurobranchaea GABA distribution was also different from that of other, more distantly related, euopisthobranch and panpulmonate snails and slugs. This suggests that the Pleurobranchaea GABA distribution may be a derived feature, unique to this lineage. The majority of GABA-ir axons and neuropil in the Nudipleura were restricted to the buccal ganglia, commissures, and connectives. However, in Tritonia and Pleurobranchaea, we detected a few GABA-ir fibers in buccal nerves that innervate feeding muscles. Although the specific functions of the GABA-ir neurons in the species in this study are not known, the innervation pattern suggests these neurons may play an integrative or regulatory role in bilaterally coordinated behaviors in the Nudipleura.


The correlation between basal serum follicle-stimulating hormone levels before embryo cryopreservation and the clinical outcome of frozen embryo transfers.
To evaluate the correlation between basal serum FSH level before the fresh IVF/intracytoplasmic sperm injection (ICSI) cycle and the clinical outcome of the subsequent frozen embryo replacement cycles. Retrospective observational study. University tertiary referral center, London, United Kingdom. Five hundred four consecutive frozen embryo transfer (FET) cycles where serum FSH levels were obtained, on days 1-4 of the cycle before the fresh IVF +/- ICSI cycles. Frozen-thawed embryo transfer. Clinical pregnancy (CP) and live birth (LB). Basal serum FSH in 127 women (25.2%) who had a CP was significantly lower compared with that in women who did not have a CP. Multivariate regression analysis showed significant correlation between basal serum FSH levels and clinical pregnancy and a low significance to LB, but there was no statistical significant differences between women who had a CP and those who did not with regard to age, treatment protocol (natural or hormone treatment cycle), or the freeze-thaw interval. The LB rate was higher in natural cycles (n = 71; 21.2%) than in hormone treatment cycles (n = 28; 16.7%). Conceiving in the fresh cycle had a positive influence on the FET outcome. Basal serum FSH level before fresh IVF/ICSI cycle is inversely correlated to a CP outcome in FET cycles. A trend was present between FSH levels and LB, but this failed to reach statistical significance.


Xeno-free workflow exhibits comparable efficiency and quality of keratinocytes isolated from human skin biopsies.
Regenerative solutions of the skin represent a hope for burn victims with extensive skin loss and chronic wound patients. The development of xeno-free workflow is crucial for clinical application in compliance with the directives of the European Medicines Agency. This study aimed at evaluating the outcome of the xeno-free isolation workflow of keratinocytes from human skin biopsy. Skin biopsies were obtained from volunteers. The epidermis was digested with TrypLE™ Select, which was deactivated by dilution or with trypsin, deactivated by media with fetal bovine serum. Freshly isolated cells were compared for total cell number, viability, activity of caspase 3, gene expression and the presence of the keratinocyte surface markers cytokeratin 14. The cells were cultured in xeno-free conditions for one week and characterized regarding the number and viability as well as the metalloproteinase secretion. The number of obtained cells was similar in both workflows. The cell viability was less in the TrypLE group, with slight reduction of the cell surface marker cytokeratin 14. Caspase 3 activity was comparable as well as the gene expression of the apoptotic markers BAX, BCL2 and SLUG, as well as the keratinocyte markers cytokeratin 14, stratifin and filaggrin. Upon culture, the number of keratinocytes, their viability and secretion of matrix metalloproteinases 1 and 10 were equal in both groups. This study reports the possibility of isolating functioning and viable keratinocytes through a xeno-free workflow for clinical application.


Human tau mutations in cerebral organoids induce a progressive dyshomeostasis of cholesterol.
Mutations in the MAPT gene that encodes tau lead to frontotemporal dementia (FTD) with pathology evident in both cerebral neurons and glia. Human cerebral organoids (hCOs) from individuals harboring pathogenic tau mutations can reveal the earliest downstream effects on molecular pathways within a developmental context, generating interacting neurons and glia. We found that in hCOs carrying the V337M and R406W tau mutations, the cholesterol biosynthesis pathway in astrocytes was the top upregulated gene set compared with isogenic controls by single-cell RNA sequencing (scRNA-seq). The 15 upregulated genes included HMGCR, ACAT2, STARD4, LDLR, and SREBF2. This result was confirmed in a homozygous R406W mutant cell line by immunostaining and sterol measurements. Cholesterol abundance in the brain is tightly regulated by efflux and cholesterol biosynthetic enzyme levels in astrocytes, and dysregulation can cause aberrant phosphorylation of tau. Our findings suggest that cholesterol dyshomeostasis is an early event in the etiology of neurodegeneration caused by tau mutations.


The clinical use of the dexamethasone suppression test in DSM-III affective disorders: correlation with the severe depressive subtypes of melancholia and psychosis.
The utility of the dexamethasone suppression test (DST) as an adjunct in the diagnosis of major depression remains controversial. While the research utility of the DST has been confirmed, the clinical utility has been questioned. We studied 166 consecutive admissions to a general, non-research unit who either met DSM-III criteria for major depression of had depressive symptoms associated with other DSM-III diagnoses. Using a 5 micrograms/dl criterion, non-suppression of serum cortisol after dexamethasone was observed in 63% of patients with DSM-III major depression. Patients with the most severe subtypes of major depression (melancholia and psychosis) showed both the highest rate of serum cortisol non-suppression and the highest post-DST serum cortisol concentrations. These findings from the clinical setting where the test, if found useful, will be used ultimately suggest that the DST is both sensitive and specific for the diagnosis of major depression. Future research will determine the potential role of the DST as an adjunct to the clinical assessment and management of patients with major affective disorder.


Effectiveness of cefotaxime alone and in combination with desacetylcefotaxime against Bacteroides fragilis.
The synergistic activity of the combination cefotaxime-desacetylcefotaxime (CTX/dCTX) was compared to the effectiveness of seven other antimicrobial agents: cefoxitin (CFOX), cefotetan (CTAN), ceftizoxime (CTIZ), chloramphenicol (CLOR), clindamycin (CLIND), metronidazole (METR), and ampicillin-sulbactam (A/S) tested against 100 clinical isolates belonging to the Bacteroides fragilis group. All tests were performed using the NCCLS reference agar-dilution method. The overall susceptibility of these organisms to CTX/dCTX was 84% compared to CFOX at 78% or CTAN at 66%. The other antimicrobials inhibited greater than 90% of these isolates. There was no difference between the susceptibility rates of CTX/dCTX and CTX with the B fragilis (85%) or B. distasonis (75%) strains. Bacteroides thetaiotaomicron showed a 11% greater susceptibility to CTX/dCTX than to CTX. Of the 100 isolates tested, 40% showed either synergy or partial synergistic interactions between CTX and dCTX. Most of the isolates showed indifference (52%), while 8% demonstrated antagonism; a relatively unique finding to date.


Sex differences in the association of urinary bisphenol-A concentration with selected indices of glucose homeostasis among U.S. adults.
Emerging evidence suggests that exposure to endocrine disruptors may initiate or exacerbate adiposity and associated health problems. This study examined sex differences in the association of urinary level of bisphenol-A (BPA) with selected indices of glucose homeostasis among U.S. adults. Data analyses were performed using a sample of 1586 participants from the 2005 to 2008 National Health and Nutrition Examination Surveys. BPA level and the ratio of BPA-to-creatinine level were defined as log-transformed variables and in quartiles. Selected indices of glucose homeostasis were defined using fasting glucose and insulin data. Multivariate linear and logistic regression models for the hypothesized relationships were constructed after controlling for age, sex, race, education, marital status, smoking status, physical activity, total dietary intake, and urinary creatinine concentration. Taking the first quartile as a referent, the third quartile of BPA level was positively associated with log-transformed level of insulin and β-cell function (homeostasis model assessment for β-cell function) as well as insulin resistance (log-transformed homeostasis model assessment for insulin resistance; homeostasis model assessment for insulin resistance ≥2.5), with significant BPA-by-sex interaction; these associations were stronger among males than among females. Irrespective of sex, the ratio of BPA-to-creatinine level was not predictive of indices of glucose homeostasis. A complex association may exist between BPA and hyperinsulinemia among adult U.S. men. Prospective cohort studies are needed to further elucidate endocrine disruptors as determinants of adiposity-related disturbances.


Teaching a child with autism to mand for information using "how".
Children with autism often do not learn to mand for information without structured teaching. Studies have demonstrated that manipulation of establishing operations (EOs), prompts, prompt fading, and differential reinforcement are effective in teaching children with autism to ask "wh" questions such as "what," "who," and "where." To date, no studies have evaluated procedures to teach children with autism to mand for information using "how." Teaching the mand, "how" is uniquely challenging because once the information regarding how to do something is provided, the EO may no longer be present. The following study evaluated a procedure to teach one child with autism to mand for information using "how" to obtain information to complete multiple activities. The results have implications for clinical application and future research on contriving EOs to teach the mand, "how."


Utility of Iodine-131 hybrid SPECT-CT fusion imaging before high-dose radioiodine therapy in papillary thyroid carcinoma.
The management protocol for differentiated thyroid cancer includes whole body iodine-131 imaging, to detect residual thyroid tissue and distant metastasis, after thyroidectomy. However, the diagnostic dose of radioiodine may fail to detect the non-functioning or poorly functioning metastasis. We present a case where hybrid single photon-emission computed tomographic and computed tomographic (SPECT-CT) fusion imaging, using a diagnostic dose of iodine-131, was able to detect both functioning as well as non-functioning pulmonary metastases, prior to high-dose radioiodine therapy.


Crohn's disease: increased mortality 10 years after diagnosis in a Europe-wide population based cohort.
No previous correlation between phenotype at diagnosis of Crohn's disease (CD) and mortality has been performed. We assessed the predictive value of phenotype at diagnosis on overall and disease related mortality in a European cohort of CD patients. Overall and disease related mortality were recorded 10 years after diagnosis in a prospectively assembled, uniformly diagnosed European population based inception cohort of 380 CD patients diagnosed between 1991 and 1993. Standardised mortality ratios (SMRs) were calculated for geographic and phenotypic subgroups at diagnosis. Thirty seven deaths were observed in the entire cohort whereas 21.5 deaths were expected (SMR 1.85 (95% CI 1.30-2.55)). Mortality risk was significantly increased in both females (SMR 1.93 (95% CI 1.10-3.14)) and males (SMR 1.79 (95% CI 1.11-2.73)). Patients from northern European centres had a significant overall increased mortality risk (SMR 2.04 (95% CI 1.32-3.01)) whereas a tendency towards increased overall mortality risk was also observed in the south (SMR 1.55 (95% CI 0.80-2.70)). Mortality risk was increased in patients with colonic disease location and with inflammatory disease behaviour at diagnosis. Mortality risk was also increased in the age group above 40 years at diagnosis for both total and CD related causes. Excess mortality was mainly due to gastrointestinal causes that were related to CD. This European multinational population based study revealed an increased overall mortality risk in CD patients 10 years after diagnosis, and age above 40 years at diagnosis was found to be the sole factor associated with increased mortality risk.


The spatiotemporal electrogram dispersion ablation targeting rotors is more effective for elderly patients than non-elderly population.
Modulating atrial fibrillation (AF) drivers has been proposed as one of the effective ablation strategies for non-paroxysmal AF (non-PAF). However, the optimal non-PAF ablation strategy is still under debate because the exact mechanisms of AF persistence including focal activity and/or rotational activity, are not well-understood. Recently, spatiotemporal electrogram dispersion (STED) assumed to indicate rotors in the form of rotational activity is proposed as an effective target for non-PAF ablation. We aimed to clarify the effectiveness of STED ablation for modulating AF drivers. STED ablation plus pulmonary vein isolation was applied in 161 consecutive non-PAF patients not undergoing previous ablation. STED areas within the entire left and right atria were identified and ablated during AF. After the procedures, the STED ablation's acute and long-term outcomes were investigated. (1) Despite a more effective acute outcome of the STED ablation for both AF termination and non-inducibility of atrial tachyarrhythmias (ATAs), Kaplan-Meier curves showed that the 24-month freedom ratio from ATAs was 49%, which resulted from the higher recurrence ratio of atrial tachycardia (AT) rather than AF. (2) A multivariate analysis showed that the determinant of ATA recurrences was only a non-elderly age, not long-standing persistent AF, and an enlarged left atrium, which were conventionally considered as key factors. STED ablation targeting rotors was effective in elderly non-PAF patients. Therefore, the main mechanism of AF persistency and the component of the fibrillatory conduction might vary between elders and non-elders. However, we should be careful about post-ablation ATs following substrate modification.


Mice lacking mitochondrial ferritin are more sensitive to doxorubicin-mediated cardiotoxicity.
Mitochondrial ferritin is a functional ferritin that localizes in the mitochondria. It is expressed in the testis, heart, brain, and cells with active respiratory activity. Its overexpression in cultured cells protected against oxidative damage and reduced cytosolic iron availability. However, no overt phenotype was described in mice with inactivation of the FtMt gene. Here, we used the doxorubicin model of cardiac injury in a novel strain of FtMt-null mice to investigate the antioxidant role of FtMt. These mice did not show any evident phenotype, but after acute treatment to doxorubicin, they showed enhanced mortality and altered heart morphology with fibril disorganization and severe mitochondrial damage. Signs of mitochondrial damage were present also in mock-treated FtMt(-/-) mice. The hearts of saline- and doxorubicin-treated FtMt(-/-) mice had higher thiobarbituric acid reactive substance levels, heme oxygenase 1 expression, and protein oxidation, but did not differ from FtMt(+/+) in the cardiac damage marker B-type natriuretic peptide (BNP), ATP levels, and apoptosis. However, the autophagy marker LC3 was activated. The results show that the absence of FtMt, which is highly expressed in the heart, increases the sensitivity of heart mitochondria to the toxicity of doxorubicin. This study represents the first in vivo evidence of the antioxidant role of FtMt. Mitochondrial ferritin (FtMt) expressed in the heart has a protective antioxidant role. Acute treatment with doxorubicin caused the death of all FtMt(-/-) and only of 60 % FtMt(+/+) mice. The hearts of FtMt(-/-) mice showed fibril disorganization and mitochondrial damage. Markers of oxidative damage and autophagy were increased in FtMt(-/-) hearts. This is the first in vivo evidence of the antioxidant role of FtMt.


Outcome of an outreach microsurgical project in rural Nepal.
This study is aimed at evaluation of the outcome of an outreach program conducted in the remote Himalayan part of Nepal. Conclusions will be used to improve blindness prevention projects in this country. A temporary outreach microsurgical clinic was established in Diktel, Nepal. All patients who reported to the clinic were examined. Cases requiring surgical treatment, mostly cataracts, were operated on. Minor ocular diseases were treated on the spot, while the complicated ones were diagnosed, given initial treatment and referred to the nearest eye hospital. Examination was performed on 1296 patients; 84 subjects underwent manual small incision cataract surgery with intraocular lens implantation. Intraoperative complications included 1 posterior capsule rupture (1.2%). Six weeks following the treatment 98.6% of the operated patients achieved corrected distance visual acuity of 0.3 or better. Out of all 1296 examined individuals, 224 (17.3%) presented visual impairment; 634 (24.5%) of the total number of eyes presented visual acuity of <0.3. In 90.2% of eyes the determinant of loss of sight was avoidable. High percentage of avoidable blindness, with the majority caused by cataract and refractive errors alone, indicate that appropriate programs focused on these diseases should be implemented. High volume cataract surgery in a camp setting with the application of appropriate surgical techniques and standardized protocols of disinfection and sterilization do not have to compromise on the quality of the outcome.


Reduced glutathione in Chinese hamster ovary cells protects against inactivation of 3-hydroxy-3-methylglutaryl coenzyme A reductase by 2-mercaptoethanol disulfide.
When the disulfide of 2-mercaptoethanol (ESSE) is added to the medium of cultured Chinese hamster ovary (CHO) cells, a time and concentration dependent release of 2-mercaptoethanol to the medium is observed. The reduction of ESSE to 2-mercaptoethanol by cells is a saturable process, the rate being approximately 50 nmoles of 2-mercaptoethanol per mg cell protein for an hour upon exposure to 250 microM ESSE. Reduction rate of ESSE by cells attached to a substratum is independent of glucose and insulin for periods up to 4 hours. However, in detached cells, swirled in suspension, addition of glucose and insulin is necessary in order to obtain a linear reduction rate of ESSE. The rate limiting enzyme in the sterol biosynthetic pathway, 3-hydroxy-3-methyl-glutaryl Coenzyme A reductase (E.C. 220.127.116.11), is inhibited by ESSE when isolated from CHO cells but total nonsaponifiable lipids synthesis from [2-14C]-acetate in intact cells is not affected by ESSE at concentrations up to 500 microM. Cytosolic reduced glutathione can spontaneously exchange disulfide bonds with ESSE and thus prevent it from inhibiting the reductase. Cultured cells respond to ESSE administration by elevating their total and acid-soluble glutathione levels. The use of ESSE as a perturbant of the GSH Status in cells is discussed.


Effect of cyclooxygenase inhibitors on pentylenetetrazol (PTZ)-induced convulsions: Possible mechanism of action.
Cyclooxygenase (COX) is reported to play a significant role in neurodegenerative and neuropsychiatric disorders, and may play a significant role in the pathogenesis of epilepsy. Various neurotransmitter abnormalities, especially of GABA and glutamate, have been reported to play a key role in the pathophysiology of epilepsy. The objective of the present study was to elucidate the effect of cyclooxygenase inhibitors on pentylenetetrazol (PTZ)-induced (80 mg/kg) convulsions in mice with possible mechanism of action. Various COX-inhibitors were administered 45 min prior to the PTZ administration. Onset, duration of clonic convulsions and percentage mortality/recovery were recorded. Pretreatment with COX-inhibitors aspirin (10 and 20 mg/kg, p.o.), naproxen (7 and 14 mg/kg, p.o.), nimesulide (1-5 mg/kg, p.o.) or rofecoxib (1-4 mg/kg, p.o.) dose-dependently showed protection against PTZ-induced convulsions. COX-2 inhibitors were more effective as compared to non-selective COX-inhibitors. Rofecoxib (1 mg/kg) or nimesulide (1 mg/kg) also enhanced the sub-protective effect of diazepam or muscimol showing GABAergic modulation of COX-2 inhibitors. COX-2 inhibitors also antagonized the effect of flumazenil (4 mg/kg)- against PTZ-induced convulsions further confirming the GABAergic mechanism. In conclusion, the results of the present study strongly suggest the possible role of cyclooxygenase isoenzymes in the pathophysiology of epilepsy and the use of COX-inhibitors as an adjuvant therapy in the treatment of epilepsy.


MBP-1 is efficiently encoded by an alternative transcript of the ENO1 gene but post-translationally regulated by proteasome-dependent protein turnover.
The c-myc promoter-binding protein-1 (MBP-1) is a transcriptional suppressor of tumorigenesis and thought to be the product of alternative translation initiation of the α-enolase (ENO1) transcript. In the present study, we cloned a 2552-bp novel cDNA with a putative coding sequence of MBP-1 and functionally examined its ability to encode the MBP-1 protein. Similarly to ENO1, the obtained MBP-1 was widely and differentially expressed in a variety of normal tissues and cancer cells. Experiments using MBP-1 promoter-driven luciferase reporter assays, biochemical cell fractionation followed by RT-PCR detection of the cytoplasmic mRNA, and transcription/translation-coupled reactions, consistently demonstrated that this novel transcript was alternatively transcribed from intron III of the ENO1 gene and was feasible for MBP-1 production. Hypoxia treatments significantly increased the transcriptional activation of the MBP-1 gene. Blocking the proteasomal degradation by MG132 stabilized the MBP-1 protein in cells. Compared with the translation efficiency for production of the MBP-1 protein, the MBP-1 transcript was 17.8 times more efficient than the ENO1 transcript. Thus, we suggest that this newly discovered transcript is a genuine template for the protein synthesis of MBP-1 in cells, and optimal expression of this gene in tumors may lead to effective clinical therapies for cancers.


Incidental diffuse low-grade gliomas: from early detection to preventive neuro-oncological surgery.
Although a large amount of data supports early surgical resection for symptomatic diffuse low-grade glioma, the therapeutic strategy is still a matter of debate regarding incidentally discovered diffuse low-grade glioma. Indeed, early and "preventive" surgery has recently been proposed in asymptomatic patients with silent diffuse low-grade glioma with better outcomes. The present review discusses the importance of an early diagnosis and of a preventive surgical treatment to improve the outcomes of incidental diffuse low-grade glioma and suggests the possible relevance of a tailored screening policy.


A new combined computational and NMR-spectroscopical strategy for the identification of additional conformational constraints of the bound ligand in an aprotic solvent.
This study documents the feasibility of switching to an aprotic medium in sugar receptor research. The solvent change offers additional insights into mechanistic details of receptor--carbohydrate ligand interactions. If a receptor retained binding capacity in an aprotic medium, solvent-exchangeable protons of the ligand would not undergo transfer and could act as additional sensors, thus improving the level of reliability in conformational analysis. To probe this possibility, we first focused on hevein, the smallest lectin found in nature. The NMR-spectroscopic measurements verified complexation, albeit with progressively reduced affinity by more than 1.5 orders of magnitude, in mixtures of up to 50% dimethyl sulfoxide (DMSO). Since hevein lacks the compact beta-strand arrangement of other sugar receptors, such a structural motif may confer enhanced resistance to solvent exchange. Two settings of solid-phase activity assays proved this assumption for three types of alpha- and/or beta-galactoside-binding proteins, that is, a human immunoglobulin G (IgG) subfraction, the mistletoe lectin, and a member of the galectin family of animal lectins. Computer-assisted calculations and NMR experiments also revealed no conspicuous impact of the solvent on the conformational properties of the tested ligands. To define all possible nuclear Overhauser effect (NOE) contacts in a certain conformation and to predict involvement of exchangeable protons, we established a new screening protocol applicable during a given molecular dynamics (MD) trajectory and calculated population densities of distinct contacts. Experimentally, transferred NOE (tr-NOE) experiments with IgG molecules and the disaccharide Gal'alpha1-3Galbeta1-R in DMSO as solvent disclosed that such an additional crosspeak, that is, Gal'OH2--GalOH4, was even detectable for the bound ligand under conditions in which spin diffusion effects are suppressed. Further measurements with the plant lectin and galectins confirmed line broadening of ligand signals and gave access to characteristic crosspeaks in the aprotic solvent and its mixtures with water. Our combined biochemical, computational, and NMR-spectroscopical strategy is expected to contribute notably to the precise elucidation of the geometry of ligands bound to compactly folded sugar receptors and of the role of water molecules in protein--ligand (carbohydrate) recognition, with relevance to areas beyond the glycosciences.


Timing the transfer of responsibilities for anaphylaxis recognition and use of an epinephrine auto-injector from adults to children and teenagers: pediatric allergists' perspective.
The optimal time for transferring responsibilities for anaphylaxis recognition and epinephrine auto-injector use from adults to children and teenagers has not yet been defined. To determine whether pediatric allergists have age-specific goals for beginning to transfer responsibilities for anaphylaxis recognition and epinephrine auto-injector use from parents and caregivers to children and teenagers at risk of anaphylaxis in the community. Members of the American Academy of Pediatrics Section on Allergy and Immunology (AAP-SOAI) were surveyed about when they typically begin to transfer these responsibilities from adults to children and teenagers. Eighty-eight allergists responded to the survey, 97.7% of whom provided service to children and teenagers with food allergies. Few allergists expected to begin transferring responsibilities for anaphylaxis recognition and epinephrine auto-injector use to children younger than 9 to 11 years. By the time their patients reached age 12 to 14 years, however, most allergists expected them to be able to describe some anaphylaxis symptoms (95.4%), demonstrate how to use an epinephrine auto-injector trainer (93.1%), begin carrying self-injectable epinephrine (88.2%), recognize the need for epinephrine (88.1%), learn to self-inject epinephrine (84.5%), and be able to self-inject epinephrine (78.6%) (cumulative data). The allergists rated the following as "very important" readiness factors for beginning to transfer responsibilities: medical history, developmental level, and ability to demonstrate auto-injector technique. Most pediatric allergists expected that by age 12 to 14 years, their patients should begin to share responsibilities with adults for anaphylaxis recognition and epinephrine auto-injector use; however, they individualized the timing based on assessment of patient readiness factors.


Sandbathing is thermoregulatory in the Mongolian gerbil, Meriones unguiculatus.
Three experiments assessed the importance of behavioral regulation of hair lipids for thermoregulation in Meriones unguiculatus. Experiment I demonstrated that animals exposed for 8-11 days at 34 degrees C had reduced body-hair lipids and a smaller increase in body temperature when exposed to tungsten light radiation than animals exposed to 5 degrees C. Experiment II confirmed that the effect was due to differential hair lipids and not heat acclimation, as the hyperthermic response and change in hair lipids were attenuated when animals were allowed to sandbathe at 34 degrees C. Experiment III showed that animals tested periodically at 30 degrees C sandbathe more frequently than those tested at 5 degrees C. Body-heat regulation is associated with variations in hair lipids and associated changes in coat color and the ability to reflect or absorb radiant energy (Thiessen, Pendergrass, & Harriman (1982) Journal of Thermal Biology, 7, 51-66). Sandbathing is temperature dependent and apparently mediates thermoregulatory changes in heat absorption and reflectance.


Heart rate variability and inflammatory bowel disease in humans: A systematic review and meta-analysis.
The autonomic nervous system (ANS) maintains homeostasis in the gastrointestinal tract, including immunity, inflammation and motility, through the brain-gut axis. To date, the associations between ANS function and inflammatory bowel disease (IBD) have been controversial and inconclusive in human studies. PubMed, Cochrane Library, and Embase were searched through February 2020 for articles reporting these association between heart rate variability (HRV), an indirect measure of ANS activity, and IBD. The standardized mean differences and 95% confidence intervals (CIs) were calculated. Ten eligible studies involving 273 ulcerative colitis patients, 167 Crohn's disease patients and 208 healthy controls were included. The values of the total power (SMD = -0.83, 95% CI = -1.44, -0.21), high frequency (SMD = -0.79, 95% CI = -1.20, -0.38), RR interval (SMD = -0.66, 95% CI = -1.04, -0.27), standard deviation of the RR intervals (SMD = -1.00, 95% CI = -1.73, -0.27), percentage of RR intervals with a greater than 50-millisecond variation (SMD = -0.82, 95% CI = -1.33, -0.30) and the square root of the mean squared differences in successive RR intervals (SMD = -0.71, 95% CI = -1.15, -0.26) of the IBD patients were lower than those of the healthy controls, and moderate to large effect sizes were observed in all HRV indices, except for low frequency (SMD = -0.41, 95% CI = 0.95, 0.13). IBD was strongly associated with an overall decrease in HRV, indicating substantially decreased ANS activity. Furthermore, the parasympathetic nerve displayed a stronger inverse association with ANS activity than the sympathetic nerve, indicating ANS dysfunction in patients with IBD.


Cut-off points for Polish-language versions of depression screening tools among patients with Type 2 diabetes.
Screening tools can help improve the detection of depression in patients with diabetes, yet the psychometric properties of most translations of scales, which are originally published in English, have not been assessed. Thus we studied the screening performance of widely used depression measures. We applied the cut-off points of the English-language versions of the Beck Depression Inventory (BDI); Hospital Anxiety and Depression Scale (HADS); Depression in Diabetes Self-Rating Scale (DDS-RS); Brief Self-Rating Scale of Depression and Anxiety (BS-RSDA); and Problematic Areas in Diabetes Survey (PAID), all of which were used to assess diabetes-specific distress in a sample of 101 patients with Type 2 diabetes. The Mini International Neuropsychiatric Interview and the Hamilton Depression Rating Scale (HDRS) were used to diagnose depression. When the English cut-off points were used, the tools had varied values of both sensitivity and specificity. When the best cut-off points were used, all measures had either very good or good sensitivity. Taking into account their length, the HADS and the BS-RSDA seem to be the best among the screening tools. Effective initial diagnosis of depression in patients with diabetes during a routine medical visit requires the use of screening tools that have adequate cut-off points. The analyses presented in this article show that screening tools should be validated and the cut-off points that are used should be population-specific.


Selective interleukin-1 receptor-associated kinase 4 inhibitors for the treatment of autoimmune disorders and lymphoid malignancy.
Pathological activation of the Toll-like receptor signaling adaptor protein MYD88 underlies many autoimmune and inflammatory disease states. In the activated B cell-like (ABC) subtype of diffuse large B cell lymphoma (DLBCL), the oncogenic MYD88 L265P mutation occurs in 29% of cases, making it the most prevalent activating mutation in this malignancy. IRAK4 kinase accounts for almost all of the biological functions of MYD88, highlighting IRAK4 as a therapeutic target for diseases driven by aberrant MYD88 signaling. Using innovative structure-based drug design methodologies, we report the development of highly selective and bioavailable small molecule IRAK4 inhibitors, ND-2158 and ND-2110. These small molecules suppressed LPS-induced TNF production, alleviated collagen-induced arthritis, and blocked gout formation in mouse models. IRAK4 inhibition promoted killing of ABC DLBCL lines harboring MYD88 L265P, by down-modulating survival signals, including NF-κB and autocrine IL-6/IL-10 engagement of the JAK-STAT3 pathway. In ABC DLBCL xenograft models, IRAK4 inhibition suppressed tumor growth as a single agent, and in combination with the Bruton's tyrosine kinase (BTK) inhibitor ibrutinib or the Bcl-2 inhibitor ABT-199. Our findings support pharmacological inhibition of IRAK4 as a therapeutic strategy in autoimmune disorders, in a genetically defined population of ABC DLBCL, and possibly other malignancies dependent on aberrant MYD88 signaling.


Involvement of the GABAergic system in the anxiolytic-like effect of the flavonoid ellagic acid in mice.
Anxiolytic-like effects of dietary flavonoids are relatively well known. Ellagic acid is a naturally occurring flavonoid compound which is abundant in many plants and fruits. The present study was designed to investigate the antianxiety-like effect of ellagic acid in mice using an elevated plus-maze test. The involvement of the GABAergic and serotonergic systems in the antianxiety-like activity of ellagic acid was also studied. Our results showed that ellagic acid treatment (25, 50 and 100 mg/kg, p.o.), produced a significant increase in the percentage of time spent and entry into the open arms, with a profile comparable to that of diazepam (1 mg/kg, p.o.). Unlike diazepam, the anxiolytic doses of ellagic acid did not prolong the duration of sodium thiopental-induced loss of righting reflex, indicating that this flavonoid is non-hypnotic. The anxiolytic effect observed with ellagic acid treatment (25 mg/kg, p.o.) was antagonized by pretreatment with picrotoxin (a non-competitive GABAA receptor antagonist, 1 mg/kg, i.p.) and flumazenil (a benzodiazepine site antagonist, 1 mg/kg, i.p.) but not with p-chlorophenylalanine (a serotonin synthesis inhibitor, 100 mg/kg, i.p.) and pindolol (a β-adrenoceptors blocker/5-HT1A/1B receptor antagonist, 10 mg/kg, i.p.). Taken together, the data demonstrated that acute and chronic administration of ellagic acid to mice has produced antianxiety-like effect when tested in the elevated plus-maze. The experiments with different receptor blockers suggest an involvement of GABAergic system in the anxiolytic action of this bioflavonoid. However, this action is not seems to be mediated through serotonergic system.


Anti-glaucoma agents-induced pseudodendritic keratitis presumed to be herpetic simplex keratitis: a clinical case series.
Anti-glaucoma agents-induced corneal toxicity may be misdiagnosed as herpetic simplex keratitis (HSK). In our study, nineteen glaucoma patients were presumed to have HSK before referral. Corneal lesions were classified into (I) linear pseudodendritic lesions formed by elevated opacified cells, (II) linear pseudodendritic lesions formed by grouped superficial punctate keratitis (SPK), (III) satellite full-thickness epithelial defects, (IV) satellite lesions formed by elevated opacified cells, and (V) geographic lesions formed by grouped SPK. We observed thirty-one events, with 15 in the lower and 16 in the central corneas. There were 21 (67.7%) type II, five (16.1%) type V, two (6.5%) of each for types III and IV, and one (3.2%) type I events. Among linear lesions (types I and II), 17 (77.3%) had horizontal and 5 (22.7%) had curvilinear orientations. Exposure duration to the last-added anti-glaucoma agent was three days to 14.5 years. About half of the events (16/31, 51.6%) used prostaglandin analogues, and 30/31 (96.8%) applied benzalkonium chloride (BAK)-containing agents. All lesions resolved within two months after decreasing offending medications or enhancing protection of ocular surface. In conclusion, anti-glaucoma agents-induced pseudodendritic keratitis presents majorly in central-lower cornea as horizontally linear lesions, and BAK-containing agents are observed in the most events.


Pharmacological heterogeneity among calcium channels that subserve acetylcholine release in vertebrate forebrain.
Inhibition of calcium-evoked [3H]ACh release by different classes of calcium channel blockers was compared among calcium-naive synaptosomes from chick, frog and rat forebrain tissues. In all three species, [3H]ACh release was insensitive to nifedipine (0.03-3 microM) but was completely inhibited by cadmium (IC50 range = 0.7-1.7 microM) or cobalt (190-350 microM). In contrast, the peptide omega-conotoxin revealed marked species heterogeneity in that [3H]ACh release was potently blocked in chick and rat synaptosomes (IC50 congruent to 1 nM), whereas frog tissue was notably resistant (IC50 greater than 100 nM). Together, these data provide functional evidence for pharmacological heterogeneity among presynaptic calcium channels that subserve [3H]ACh release.


Image findings of a false positive radioactive iodine-131 uptake mimicking metastasis in pulmonary aspergillosis identified on single photon emission computed tomography-computed tomography.
High doses of iodine-131 are commonly used in patients with differentiated thyroid cancer after total or subtotal thyroidectomy, in order to ablate the remaining cancer or normal thyroid tissue. Multiple different false-positive scans can occur in the absence of residual thyroid tissue or metastases. The authors present a case of abnormal uptake of radioactive iodine in the aspergilloma, potentially masquerading as pulmonary metastases.


Impact of skip metastasis in gastric cancer.
Several studies have shown the features of skip metastasis in other cancers besides gastric cancer. Since minimally invasive surgery has been applied to gastric cancer, the concerns and awareness of skip metastasis have grown in the medical community. We conducted the present retrospective study to reveal the clinicopathological characteristics of patients with skip metastasis. We also wished to clarify the clinical impact of skip metastasis for gastric cancer. Five hundred and eighty-nine patients having lymphatic metastases were enrolled in the present study. Among them, 266 patients had positive nodes extending into the N2 group. We divided these patients into the skip positive (+) and the skip negative (-) group, and we comparatively analysed clinicopathological factors and calculated the survival probabilities for the two groups. The skip (+) and skip (-) groups involved 14 (5.3%) and 252 (94.7%) patients, respectively. Of all the investigated factors, a significant difference between two groups was observed only in the total number of retrieved nodes. Stations of skip nodes were along left gastric (7), anterior common hepatic (8a), celiac (9), splenic (11) artery and right paracardial nodes (1). The survival curves calculated in the present study did not show any statistical differences between the groups. Due to problems of skip metastasis in gastric cancer, D2 lymph node dissection should be performed until sentinel node detection is feasible and reliable. The potential risk from skip metastasis is not great and skip metastasis itself should not be a major consideration in therapeutic decisions.


Photoredox Generation of Carbon-Centered Radicals Enables the Construction of 1,1-Difluoroalkene Carbonyl Mimics.
Described is a facile, scalable route to access functional-group-rich gem-difluoroalkenes. Using visible-light-activated catalysts in conjunction with an arsenal of carbon-radical precursors, an array of trifluoromethyl-substituted alkenes undergoes radical defluorinative alkylation. Nonstabilized primary, secondary, and tertiary radicals can be used to install functional groups in a convergent manner, which would otherwise be challenging by two-electron pathways. The process readily extends to other perfluoroalkyl-substituted alkenes. In addition, we report the development of an organotrifluoroborate reagent to expedite the synthesis of the requisite trifluoromethyl-substituted alkene starting materials.


A case of fatal ephedra intake associated with lipofuscin accumulation, caspase activation and cleavage of myofibrillary proteins.
Ephedra, a herb reported to suppress appetite and stimulate the sympathetic nervous system as well as cardiac performance, has recently been related to several adverse events, including seizure, stroke, hypertension, myocardial infarction, and sudden death. Here, we describe the case of a 45-year-old woman who died of cardiovascular collapse while taking ephedra. Tissue analysis revealed non-specific degenerative alterations in the myocardium (lipofuscin accumulation, basophilic degeneration and vacuolation of myocytes, as well as myofibrillary loss), associated with myocyte apoptosis, caspase activation, and extensive cleavage of miofibrillary proteins alpha-actin, alpha-actinin, and cardiac troponin T. Healthcare professionals are therefore urged to warn their patients about the risk of serious adverse effects, which may follow ephedra intake.


A glance at genome editing with CRISPR-Cas9 technology.
In recent years, CRISPR-Cas9 technology is widely acknowledged for having major applications in the field of biotechnology for editing genome of any organism to treat a variety of complex diseases and for other purposes. The acronym 'CRISPR-Cas' stands for clustered regularly interspaced short palindromic repeats-CRISPR-associated genes. This genetic organization exists in prokaryotic organisms and aids in the development of adaptive immunity since a protein called Cas9 nuclease cleaves specific target nucleic acid sequences from foreign invaders and destroys them. This mode of action has gained interest of the researchers to understand the insights of CRISPR-Cas9 technology. Here, we review that CRISPR-Cas organization is restricted to two classes and possesses different protein effectors. We also review the architecture of CRISPR loci, mechanism involved in genome editing by CRISPR-Cas9 technology and pathways of repairing double-strand breaks (DSBs) generated during the process of genome editing. This review also presents the strategies to increase the Cas9 specificity and reduce off-target activity to achieve accurate genome editing. Further, this review provides information on CRISPR tools used for genome editing, databases that are required for storing data on loci, strategies for delivering CRISPR-Cas9 to cells under study and applications of CRISPR-Cas9 to various fields. Safety measures are implemented on this technology to avoid misuse or ethical issues. We also discuss about the future aspects and potential applications of CRISPR-Cas9 technology required mainly for the treatment of dreadful diseases, crop improvement as well as genetic improvement in human.


Fungal artificial chromosomes for mining of the fungal secondary metabolome.
With thousands of fungal genomes being sequenced, each genome containing up to 70 secondary metabolite (SM) clusters 30-80 kb in size, breakthrough techniques are needed to characterize this SM wealth. Here we describe a novel system-level methodology for unbiased cloning of intact large SM clusters from a single fungal genome for one-step transformation and expression in a model host. All 56 intact SM clusters from Aspergillus terreus were individually captured in self-replicating fungal artificial chromosomes (FACs) containing both the E. coli F replicon and an Aspergillus autonomously replicating sequence (AMA1). Candidate FACs were successfully shuttled between E. coli and the heterologous expression host A. nidulans. As proof-of-concept, an A. nidulans FAC strain was characterized in a novel liquid chromatography-high resolution mass spectrometry (LC-HRMS) and data analysis pipeline, leading to the discovery of the A. terreus astechrome biosynthetic machinery. The method we present can be used to capture the entire set of intact SM gene clusters and/or pathways from fungal species for heterologous expression in A. nidulans and natural product discovery.


Effect of preradiation dental intervention on incidence of osteoradionecrosis in patients with head and neck cancer: A systematic review and meta-analysis.
The purpose of this systematic review was to examine whether dental intervention involving bone or soft-tissue manipulation preradiotherapy (pre-RT) is associated with lower rates of osteoradionecrosis of the jaw (ORNJ) in patients with head and neck cancer (HNC). The authors included relevant studies from MEDLINE, Embase, and Cochrane Library, including observational studies published from 2007 through 2021 and involving adults who underwent dental intervention pre-RT for HNC. Authors assessed evidence certainty by using the Grading of Recommendations Assessment, Development, and Evaluation approach. Random-effects models were used to calculate pooled relative risk estimates and hazard ratios. When meta-analysis was not possible, study-level measures of association and narrative summaries of the evidence were reported. Twenty-two studies were included. From the pooled, unadjusted analysis, patients undergoing pre-RT extractions may have a 55% increased risk of experiencing ORNJ (relative risk, 1.55; 95% CI, 0.85 to 2.86; very low certainty); the unadjusted pooled hazard ratio was 3.19 (95% CI, 0.99 to 10.31; very low certainty), corresponding to a possible increased hazard of developing ORNJ (very low certainty). Findings for other pre-RT procedures manipulating bone or tissue relied on limited, observational studies with low or very low certainty evidence. Mostly very low certainty evidence suggests that patients with HNC who need pre-RT dental intervention may have an increased risk of developing ORNJ compared with those who do not. Maintaining optimal oral health may help reduce the need for urgent pre-RT dental treatment, potentially reducing ORNJ risk and minimizing delay of oncologic treatment in patients with HNC.


Using laws, common sense, and statistical approaches to design indicators for 'medical desertification'. An application on the Romanian case.
The study of spatial accessibility to healthcare services is key to health policy (Pförtner et al., 2019; Vergier et al., 2017). Terms such as 'rurality' or 'medical desertification' were employed to stress the relevance of the topic. Within the existing literature, there is little (if any) concern with the legitimacy of the existing ways to measure inequality. Expert systems were assumed to be in place, and discrepancies or consistency with existing regulations or to views of relevant stakeholders were not considered. This paper discusses spatial accessibility of medical services in three distinct approaches: normative, that is following what national regulations consider as standard; relative, that is what statistical approaches reveal; consensual, which adjusts the indexes based on representations of stakeholders. The three approaches are compared for the case of Romania, an EU country with low population density, ideal to inspect geographical discrepancies. For the relative and the consensual approach, population is adjusted according to different demand expressed by age groups, and also including population and providers from the nearby localities. The normative approach follows official regulations. The refinements in the consensual model are based on survey data from stakeholders and consider distances to neighbouring localities and adjustments according to the age structure of the population in the catchment area. The results reveal high consistency between the consensual and the relative approach. Both are more permissive with respect to detecting medical desertification as compared to the normative approach but prove to be more effective in directing policy when resources are scarce. The normative approach, however, is relevant in depicting the state of the system as contrasted to a desired standard. The relative approach also overlaps with the consensual one. Therefore, to fully characterize spatial accessibility to general practitioners (GPs) and pharmacies, one needs to consider at least the normative and the relative approaches.


Multidimensional evaluation method and application based on life cycle carbon efficiency considering carbon emission, cost, and function.
Low carbon research has currently become the most discussed environmental issue. Current comprehensive evaluation methods for low carbon consider carbon emission, cost, process parameters, and resource utilization, but the realization of low carbon may lead to cost fluctuations and functional changes and lack consideration of product functional requirements. Hence, this paper developed a multidimensional evaluation method for low-carbon research based on the association among three dimensions, namely, carbon emission, cost, and function. This multidimensional evaluation method was named life cycle carbon efficiency (LCCE), which is defined as the ratio between the value in the life cycle and the carbon emissions generated. The LCCE model was established, and then carbon emission calculation, cost assessment, and function quantification of the life cycle were performed according to the three dimensions. Case study and sensitivity analysis verified the feasibility of the proposed method. The method had comprehensive and accurate evaluation results, which provided theoretical support and optimized the low-carbon design.


Intracranial aneurysms in geriatric patients: angiographic features and angioautotomographic analyses.
The angiographic investigation of intracranial aneurysms in geriatric patients was made by conventional cerebral angiography and angioautomography. Analyses were based on 49 aneurysms in 38 patients aged over 60 who had 24 (49.0%) large aneurysms of maximum diameter over 100 mm. This contrasted strongly with the aneurysms of patients aged under 60, only 14.4% of which were large. Cerebral angioautotomography had a particular diagnostic value for such large aneurysms, revealing not only the stalks and the whole shapes of aneurysms, but also the normal parent arteries which were hidden on conventional cerebral angiography.


Collagen content and types in the intestinal strictures of Crohn's disease.
The collagen content and the relative amount of collagen types were quantitated in control intestine as well as in both inflamed and strictured intestine resected from patients with Crohn's disease. The major collagen type in control intestine was type I (68%), followed by types III (20%) and V (12%). In strictured intestine both collagen content and the relative amount of type V collagen were significantly increased compared with control intestine. Histologic studies demonstrated that in strictured specimens there was a striking proliferation of smooth muscle cells of the muscularis mucosae associated with an accumulation of collagen in the submucosa. The thickness of the muscularis propria was also increased. Immunohistochemical studies demonstrated small amounts of type V collagen in the submucosa of control bowel. In contrast, large amounts of type V collagen were seen in the fibrotic, expanded submucosa of strictured bowel, particularly in the areas where smooth muscle cells of the muscularis mucosae had proliferated. Intestinal strictures in Crohn's disease are therefore characterized by an accumulation of collagen, a proliferation of smooth muscle cells, and an increase in type V collagen, a collagen type produced in relatively large amounts by smooth muscle cells. These changes appear to result in both a loss of the normal compliance of the intestine and a thickening of the intestine wall, resulting ultimately in the intestinal obstruction so frequently seen in patients with Crohn's disease.


51Cr release cytotoxicity assay evaluated with fluoride and cadmium.
The sensitivity of the 51Cr release assay was evaluated with two known cytotoxic agents; fluoride and cadmium. The sensitivity was far less than that of other assays, e.g. growth inhibition of cultured cells, and additional a two-peaked dose-response curve was found with cadmium.


Nanodevices based on mesoporous glass nanoparticles enhanced with zinc and curcumin to fight infection and regenerate bone.
Nanotechnology-based approaches are emerging as promising strategies to treat different bone pathologies such as infection, osteoporosis or cancer. To this end, several types of nanoparticles are being investigated, including those based on mesoporous bioactive glasses (MGN) which exhibit exceptional structural and textural properties and whose biological behaviour can be improved by including therapeutic ions in their composition and loading them with biologically active substances. In this study, the bone regeneration capacity and antibacterial properties of MGNs in the SiO2-CaO-P2O5 system were evaluated before and after being supplemented with 2.5% or 4% ZnO and loaded with curcumin. in vitro studies with preosteoblastic cells and mesenchymal stem cells allowed determining the biocompatible MGNs concentrations range. Moreover, the bactericidal effect of MGNs with zinc and curcumin against S. aureus was demonstrated, as a significant reduction of bacterial growth was detected in both planktonic and sessile states and the degradation of a pre-formed bacterial biofilm in the presence of the nanoparticles also occurred. Finally, MC3T3-E1 preosteoblastic cells and S. aureus were co-cultured to investigate competitive colonisation between bacteria and cells in the presence of the MGNs. Preferential colonisation and survival of osteoblasts and effective inhibition of both bacterial adhesion and biofilm formation of S. aureus in the co-culture system were detected. Our study demonstrated the synergistic antibacterial effect of zinc ions combined with curcumin and the enhancement of the bone regeneration characteristics of MGNs containing zinc and curcumin to obtain systems capable of simultaneously promoting bone regeneration and controlling infection. STATEMENT OF SIGNIFICANCE: In search of a new approach to regenerate bone and fight infections, a nanodevice based on mesoporous SiO2-CaO-P2O5 glass nanoparticles enriched with Zn2+ ions and loaded with curcumin was designed. This study demonstrates the synergistic effect of the simultaneous presence of zinc ions and curcumin in the nanoparticles that significantly reduces the bacterial growth in planktonic state and is capable to degrade pre-formed S. aureus biofilms whereas the nanosystem exhibits a cytocompatible behaviour in the presence of preosteoblasts and mesenchymal stem cells. Based on these results, the designed nanocarrier represents a promising alternative for the treatment of acute and chronic infections in bone tissues, while avoiding the significant current problem of bacterial resistance to antibiotics.


Phylogeny, biogeography and character evolution in the tribe Desmodieae (Fabaceae: Papilionoideae), with special emphasis on the New Caledonian endemic genera.
The nearly cosmopolitan tribe Desmodieae (Fabaceae) includes many important genera for medicine and forage. However, the phylogenetic relationships among the infratribal groups circumscribed using morphological traits are still poorly known. In this study, we used chloroplast (rbcL, psbA-trnH) and nuclear (ITS-1) DNA sequences to investigate the molecular phylogeny and historical biogeography of Desmodieae, and infer ancestral states for several vegetative and reproductive traits. Three groups, corresponding to the Desmodium, Lespedeza, and Phyllodium groups sensu Ohashi were retrieved in the phylogenetic analyses. Conflicts in the topologies inferred from the chloroplast and nuclear datasets were detected. For instance, the Lespedeza clade was sister to the groups Phyllodium+Desmodium based on chloroplast DNA, but nested within the Desmodium group based on ITS-1. Moreover, the New Caledonian endemic genera Arthroclianthus and Nephrodesmus were not monophyletic but together formed a clade, which also included Hanslia and Ohwia based on chloroplast DNA. The hypothetical common ancestor of Desmodieae was dated to the Middle Oligocene (ca. 28.3Ma) and was likely an Asian shrub or tree producing indehiscent loments. Several colonization events towards Oceania, America, and Africa occurred (all less than ca. 17.5Ma), most probably through long distance dispersal. The fruits of Desmodieae repeatedly evolved from indehiscence to dehiscence. We also showed that indehiscent loments allow for more variability in the number of seeds per fruit than indehiscent legumes. Modularity seems here to allow variability in the number of ovules produced in a single ovary.


Association between TLR4 polymorphisms (896 A>G, 1196 C>T, - 2570 A>G, - 2081 G>A) and virulence factors in uropathogenic Escherichia coli.
Escherichia coli is the main etiological agent of urinary tract infections. Its virulence factors are important during the initial interaction stage with the host as they enable colonization of urinary tract tissues. The genetic markers evidencing susceptibility to develop recurrent infections have been previously described. Toll-like receptors are critical sensors of microbial attacks, and they are also effectors of the individual's innate defense for elimination of pathogens. The aim of this study was to evaluate the association between functional polymorphisms (896 A>G, 1196 C>T, - 2570 A>G, - 2081 G>A) and susceptibility to develop urinary tract infections as well as E. coli virulence factors. This study includes 100 samples from patients diagnosed with UTI and 100 samples from uninfected subjects. A conventional urine culture was performed and the isolates were identified by using the Vitek automated system. TLR4 gene polymorphisms were identified by the PCR-RFLP technique. The hlyA, fimH, papC, iutA and cnf1 virulence factors as well as the E. coli phylogenetic group were assessed by PCR. In this study, it was observed that the presence of the - 2570 polymorphism represents a risk of UTI (p < 0.01), whereas - 2081 confers protection (p < 0.01). The 896A>G and 1196C>T polymorphisms were associated with the E. coli virulence factors fimH and hlyA, respectively (p < 0.05). The B2 group was the most frequent in clinical isolates (51%), and it displayed more virulence factors regarding other phylogenetic groups (p ≤ 0.05). An interesting finding was that strains considered as commensals, belonging to groups A and B1, can cause UTI and present virulence factors. Polymorphisms occurring in the TLR4 promoter region are correlated with susceptibility or risk of UTI, whereas structural polymorphisms are associated with the recognition of virulence factors displayed by E. coli.


A method to measure an indicator of viraemia in Atlantic salmon using a reporter cell line.
RTG-P1 is a transgenic fish cell line producing luciferase under the control of the IFN-induced Mx rainbow trout gene promoter. This cell line was used to measure viraemia of Salmonid alphavirus (SAV), the cause of Salmon Pancreas Disease (SPD), a serious disease in farmed Atlantic salmon. Two SAV genotype 1 (SAV1) isolates were used in this study, F93-125 (tissue-culture adapted, from Ireland) and 4640 (from a field case in Scotland). The kinetics and magnitude of luciferase activity were monitored versus the time of infection. During a direct infection experiment, the induction of luciferase significantly increased 16- and 4-fold after incubation for 6 days with F93-125 at 15 and 20°C, respectively. Filtration and heat treatment experiments demonstrated that the luciferase induction in RTG-P1 was dependent on viral replication. Unlike many cell lines used in fish viral diagnostic, RTG-P1 is not sensitive to salmonid serum, therefore, viraemia could be successfully monitored on serum collected from fish during a cohabitation challenge with 4640 isolate. A peak of viraemia could be detected 16 days post IP inoculation of the shedders. This novel cost-effective method to measure viraemia does not rely on development of cytopathic effect (CPE) in culture, is compatible with non-lethal blood collections in fish and can be used to assign emerging diseases to a viral aetiology.


Implementation of transvaginal ultrasound in an emergency department residency program: an analysis of resident interpretation.
Emergency physicians are increasingly performing transvaginal ultrasound (TVUS) to rule out ectopic pregnancy. However, little is known about appropriate educational pathways to train emergency medicine residents in TVUS. To evaluate the ability of Emergency Medicine (EM) residents who underwent a training program in TVUS to detect the presence or absence of an intrauterine pregnancy (IUP) in patients of < 13 weeks gestation with vaginal bleeding or abdominal pain, as compared to the final interpretation of each study as determined by the Emergency Department (ED) Director of Ultrasound. This was a prospective, observational study in a single residency program. Training included a lecture, competency examination, and 10 supervised TVUSs. The EM residents then performed TVUSs with the goal of determining the presence or absence of an IUP without input from an attending physician. Correlation with the ED Director of Ultrasound was assessed for the cohort, and stratified by year of training. There were 22 residents who performed 75 TVUSs over 17 months. Correlation with the ED Director of Ultrasound was 93.3%. Differences in correlation with the ED Director of Ultrasound were noted when compared by year of training: post-graduate year (PGY)-3 (93.3%), PGY-2 (92.1%), and PGY-1 (100%); p < 0.001. Residents were able to perform TVUSs to determine the presence or absence of an IUP in patients in whom the diagnosis of ectopic pregnancy was being considered with a high degree of correlation with the ED Director of Ultrasound after a brief training program. Correlation with the ED director of ultrasound was influenced by year of training.


Interobserver reliability in physical examination of the cervical spine in patients with headache.
To assess interobserver reliability of two expert headache neurologists when examining the cervical spine of patients with headache. The diagnosis of cervicogenic headache involves the physical examination of the cervical spine. Twenty-four patients diagnosed as having migraine, tension-type headache, or cervicogenic headache were included in the study. After interview, each patient's cervical spine was examined in a structured way. Reliability was assessed by Cohen's kappa. Reduced range of motion in the cervical spine showed kappa scores indicating moderate agreement. Provocation of headache revealed moderate-to-substantial agreement. Assessment of zygapophyseal joint pressure pain showed slight-to-fair agreement. The kappa values of the circumscribed characteristic tender points showed agreement ranging from "not better than chance" to "substantial agreement." Our study showed that the interobserver reliability of expert headache neurologists was satisfactory in the majority of the physical examination tests of the cervical spine in patients with different headache syndromes. However, standardization of the clinical tests in order to improve their reliability is recommended.


Enhancement of murine B cell responses to lipopolysaccharide by supernatants of irradiated peripheral blood leukocytes.
At low cell density, the proliferative response of B cells to lipopolysaccharide (LPS) is not detectable. We investigated under these experimental conditions the role of several cell populations on the LPS-induced B-cell proliferation. The addition to murine B cells of irradiated peripheral blood leukocytes (PBL) from the C3H/HeJ mouse strain, or of culture supernatants of these cells, efficiently restored a response to LPS. Similar results were also obtained with irradiated PBL from other mouse strains and from rabbits. The activities of the culture supernatants were not significantly modified when the PBL were depleted of adherent cells or of Thy-1.2 positive cells, thus suggesting that the active factors were secreted neither by T cells, nor by monocytes.


Probing the association between maternal anxious attachment style and mother-child brain-to-brain coupling during passive co-viewing of visual stimuli.
Brain-to-brain coupling during co-viewing of video stimuli reflects similar intersubjective mentalisation processes. During an everyday joint activity of watching video stimuli (television shows) with her child, an anxiously attached mother's preoccupation with her child is likely to distract her from understanding the mental state of characters in the show. To test the hypothesis that reduced coupling in the medial prefrontal cortex (PFC) would be observed with increasing maternal attachment anxiety (MAA), we profiled mothers' MAA using the Attachment Style Questionnaire and used functional Near-infrared Spectroscopy (fNIRS) to assess PFC coupling in 31 mother-child dyads while they watched three 1-min animation videos together. Reduced coupling was observed with increasing MAA in the medial right PFC cluster which is implicated in mentalisation processes. This result did not survive control analyses and should be taken as preliminary. Reduced coupling between anxiously-attached mothers and their children during co-viewing could undermine quality of shared experiences.


Familial Aggregation of CKD and Heritability of Kidney Biomarkers in the General Population: The Lifelines Cohort Study.
Chronic kidney disease (CKD) has a heritable component. We aimed to quantify familial aggregation of CKD in the general population and assess the extent to which kidney traits could be explained by genetic and environmental factors. Cross-sectional 3-generation family study. Data were collected at entry into the Lifelines Cohort Study from a sample of the general population of the northern Netherlands, composed predominantly of individuals of European ancestry. Family history of CKD. The primary outcome was CKD, defined as estimated glomerular filtration rate (eGFR)<60mL/min/1.73m2, where GFR was estimated using the CKD Epidemiology Collaboration creatinine equation. Among a subsample for which urinary albumin concentration was available (n=59,943), urinary albumin excretion was expressed as the rate of urinary albumin excretion (UAE) per 24 hours or urinary albumin-creatinine ratio (UACR). Familial aggregation of CKD was assessed by calculating the recurrence risk ratio (RRR), using adapted Cox proportional hazards models. Heritability of continuous kidney-related traits was estimated using linear mixed models and defined as the ratio of the additive genetic variance to total phenotypic variance. All models were adjusted for age, sex, and known risk factors for kidney disease. Among 155,911 participants with available eGFR data, the prevalence of CKD was 1.19% (1,862 cases per 155,911). The risk of CKD in those with an affected first-degree relative was 3 timeshigher than the risk in the total sample (RRR, 3.04 [95% CI, 2.26-4.09). In those with an affected spouse, risk of CKD was also higher (RRR, 1.56 [95% CI, 1.20-1.96]), indicative of shared environmental factors and/or assortative mating. Heritability estimates of eGFR, UAE, and UACR were 44%, 20%, and 18%, respectively. For serum urea, creatinine, and uric acid, estimates were 31%, 37%, and 48%, respectively, whereas estimates for serum electrolytes ranged from 22% to 28%. Use of estimated rather than measured GFR. UAE data only available in a subsample. In this large population-based family study, a positive family history was strongly associated with increased risk of CKD. We observed moderate to high heritability of kidney traits and related biomarkers. These results indicate an important role of genetic factors in CKD risk.


Nuclear factor-kappaB activation and differential expression of survivin and Bcl-2 in human grade 2-4 astrocytomas.
Antiapoptotis resulting from hyperactivation of the transcription factor NF-kappaB has been described in several cancer types. It is triggered by the interaction of the tumor necrosis factor (TNF) with its receptors and recruitment of the intermediate factor TNF-receptor associated factor (TRAF) 2. The NF-kappaB transcriptional activity could amplify the expression of antiapoptotic genes. The authors investigated the activity of NF-kappaB, and the mRNA expression of TNFalpha, TNFalpha receptor, TRAF1, TRAF2, and TRAF-associated NF-kappaB activator (TANK), and the antiapoptotic genes Bcl-2, c-IAP 1 and 2, and Survivin in human astrocytic tumors. Eight low-grade astrocytomas (LGA), 10 anaplastic astrocytomas (AAs), 10 glioblastoma multiforme (GBM) samples were used; 4 samples of normal brain tissue were used as controls. The NF-kappaB activation was analyzed by electrophoretic mobility shift assay; TRAF1, TRAF2, TANK/I-TRAF, Bcl-2, c-IAP 1 and 2, and Survivin mRNA expressions were studied using real-time quantitative reverse-transcriptase polymerase chain reaction. NF-kappaB hyperactivity was detected in tumor samples. mRNA of antiapoptotic genes, particularly BCL-2 and Survivin, was hyperexpressed in gliomas. Interestingly, BCL-2 was hyperexpressed in LGAs, whereas a very high level of Survivin featured high-grade gliomas. The differential expression of antiapoptotic genes yielded a tight clustering of all LGA and nearly all GBM samples in cluster analysis. NF-kappaB and factors involved in its intracellular activation were up-regulated in gliomas. NF-kappaB-activated antiapoptotic genes were hyperexpressed in tumor samples, but showed a differential expression with higher levels of Bcl-2 in LGAs and higher levels of Survivin in GBMs.


Angiographic predictors of survival following left ventricular aneurysmectomy.
Sixty-four consecutive patients with coronary artery disease who had resections of left ventricular scars during 1969 to 1973 were retrospectively identified. Extent of angiographic coronary artery disease was scored by the jeopardy score system. Size of the abnormally contracting segment (akinetic or dyskinetic in all) was measured as a percent of the end-diastolic ventriculographic perimeter (% ACS). Contractility of the non-ACS was expressed as the difference between the actual ejection fraction and that predicted by the spherical model of Feild and Dowling (excess ejection fraction, XEF.) Perioperative survival correlated with jeopardy score (21 of 27, jeopardy score less than or equal to 6; 19 of 37 jeopardy score greater than 6) and with XEF (30 OF 38, XEF greater than +0.10; 10 of 26, XEF less than or equal to "0.10). When XEF and jeopardy score were combined, the patients were separated into four subgroups with perioperative survival ranging from 89% to 33%. Long-term survival (minimum follow-up period 30 months) in the 40 perioperative survivors also correlated with jeopardy score (95% at 54 mo for jeopardy score less than or equal to 6; 49% for jeopardy score greater than 6). Survival was unrelated to whether or not aortocoronary bypass graft procedures had been done. It is concluded that survival following aneurysmectomy is predicted by two preoperative angiographic variables--the extent of coronary artery disease and the contractility of the non-aneurysmal portion of the left ventricle.


The underlying mechanism of transcription factor IRF1, PRDM1, and ZNF263 involved in the regulation of NPPB rs3753581 on pulse pressure hypertension.
To investigate the correlation between NPPB gene variants and pulse pressure hypertension and the underlying regulatory mechanisms and try to confirm that NPPB may be a potential molecular target of gene therapy for pulse pressure hypertension. A total of 898 participants were recruited from the First Affiliated Hospital of Fujian Medical University and the plasmids with differential expression of NPPB were constructed. Genotype distribution of NPPB(rs3753581, rs198388, and rs198389)was analyzed and the expression of N-terminal pro-B-type natriuretic peptide(NT-proBNP) and renin-angiotensin -aldosterone system(RAAS) related indicators were identified in the groups studied. According to a genotype analysis, there was a significant difference in the genotype distribution of NPPB rs3753581 among the groups (P = 0.034). In logistic regression analysis, NPPB rs3753581 TT was associated with a 1.8-fold greater risk of pulse pressure hypertension than NPPB rs3753581 GG (odds ratio = 1.801; 95% confidence interval: 1.070-3.032; P = 0.027). The expression of NT-proBNP and RAAS related indicators in clinical and laboratory samples showed striking differences. The activity of firefly and Renilla luciferase in pGL-3-NPPB-luc (-1299G) was higher than pGL-3-NPPBmut-luc(-1299 T)(P < 0.05). The binding of NPPB gene promoter rs3753581 (-1299G) with transcription factors IRF1, PRDM1, and ZNF263 was predicted and validated by the bioinformatics software TESS and chromatin immunoprecipitation(P < 0.05). NPPB rs3753581 was correlated with genetic susceptibility to pulse pressure hypertension and the transcription factors IRF1, PRDM1, and ZNF263 may be involved in the regulation of NPPB rs3753581 promoter (-1299G) on the expression of NT-proBNP/RAAS.


Direct Tumor Killing and Immunotherapy through Anti-SerpinB9 Therapy.
Cancer therapies kill tumors either directly or indirectly by evoking immune responses and have been combined with varying levels of success. Here, we describe a paradigm to control cancer growth that is based on both direct tumor killing and the triggering of protective immunity. Genetic ablation of serine protease inhibitor SerpinB9 (Sb9) results in the death of tumor cells in a granzyme B (GrB)-dependent manner. Sb9-deficient mice exhibited protective T cell-based host immunity to tumors in association with a decline in GrB-expressing immunosuppressive cells within the tumor microenvironment (TME). Maximal protection against tumor development was observed when the tumor and host were deficient in Sb9. The therapeutic utility of Sb9 inhibition was demonstrated by the control of tumor growth, resulting in increased survival times in mice. Our studies describe a molecular target that permits a combination of tumor ablation, interference within the TME, and immunotherapy in one potential modality.


Renal leiomyoma in a child: a rare renal tumor.
Renal leiomyoma is a rare benign smooth muscle tumor of the kidney. Most cases are reported in adults, with very few cases described in children. We report a case of right renal leiomyoma in a 6-year-old boy which was suspected of being a Wilms tumor. Contrast-enhanced computed tomography could not differentiate leiomyoma in the kidney, and the mass was diagnosed as a Wilms tumor. Right radical nephrectomy was performed. Diagnosis of renal leiomyoma could only be achieved after histopathologic examination and immunohistochemistry. We recommend that total nephrectomy has to be done in most cases of pediatric renal tumors to avoid the risk of malignancy. There may be a role for nephron sparing surgery, provided the tumor is small, and one has a strong suspicion for the lesion being benign.


A novel enzymatic tool for transferring GalNAc moiety onto challenging acceptors.
The β-N-acetylhexosaminidase from Penicillium oxalicum (PoHex; EC 126.96.36.199) is a fungal glycosidase with an outstandingly high GalNAcase/GlcNAcase activity ratio. It has a remarkable synthetic capability and can process carbohydrates functionalized at various positions. However, the production in the native fungal host is lengthy, unselective and purification from the fungal medium is complicated and low yielding. We present here a novel production method of this enzyme in the eukaryotic host of Pichia pastoris, followed by elegant one-step purification to homogeneity. The resulting recombinant enzyme has improved biochemical and catalytic properties compared to the fungal wild type. Its good production yield (11 mg/400 mL cultivation medium) greatly expands the scope of synthetic applications. We further demonstrate the synthetic utility and broad acceptor specificity of recombinant PoHex in the glycosylation of a series of challenging acceptors with varying structural architectures, namely secondary and tertiary hydroxyl, aldoxime and a poly-hydroxylated compound.


Successful pregnancy in a patient suffering from recurrent mid-trimester miscarriage with C9 deficiency after receiving cervical cerclage followed by clindamycin and progesterone: a case report.
Complement component 9 (C9) deficiency is relatively common, especially in Japan. Here we present the case of a 27-year-old Japanese woman whose obstetric history involved three mid-trimester miscarriages (at 22 weeks', 18 weeks' and 21 weeks' gestation) and one early spontaneous miscarriage. Her fifth pregnancy was successfully managed by cervical cerclage at 13 weeks' gestation, followed by clindamycin administration (600 mg/day for 7 days) and progesterone injections (250 mg/week). She gave birth to a healthy 3326-g male infant at 40 weeks and 1 day gestation after natural onset of labor. After delivery, the serum complement components were analyzed. C9 protein and activity were undetectable in the patient's serum. We suggest that an immunologic disorder such as C9 deficiency should be considered as a potential complication of undiagnosed recurrent miscarriages.


A new selective medium for the isolation of Salmonellae other than Salmonella typhi.
A medium consisting of a MacConkey's base with added magnesium, calcium, cobalt, zinc, thiosulphate, novobiocin, iron dextran, Tween 80 and EDTA has been found highly selective for Salmonellaparatyphi B and most of the food poisoning salmonellae that commonly occur in Britain. Nearly all the normal faecal flora is inhibited.


Chronic cyclosporine-A injection in rats with damaged blood-brain barrier does not impair retention of passive avoidance.
Recently, we demonstrated that chronic administration of immunosuppressant drug, cyclosporine-A (CsA), does not produce impairment in memory retention of a passive avoidance task in normal adult rats. Since CsA has been used as an adjunctive therapy to avoid xenograft rejection inherent in neural transplantation therapy for neurodegenerative disorders, we replicated our previous study in animals with damaged blood-brain barrier (BBB) simulating that of the neural transplantation protocol. Adult rats with damaged BBB that received either chronic CsA (5, 10, and 20 mg/kg) or vehicle injection did not differ significantly in their memory retention of the passive avoidance task that rewarded 'less mobile activity', in that animals avoided electric shock when they restrained their movements within the safe compartment. General spontaneous locomotor activity also was not altered by CsA, except in animals that received 20 mg/kg, which displayed significant hypoactivity at later post-injection periods of CsA. The absence of potentiation of retention of the passive avoidance task in all CsA-treated animals, including the hypoactive ones, suggests that locomotor activity did not interfere with cognitive behavior. The present results confirm our previous findings that the therapeutic dosage (10 mg/kg) of CsA used for neural transplantation does not produce visible deleterious effects on the performance of memory retention task in immunosuppressed rats with damaged BBB.


Streptococcus anginosus group infections: Management and outcome at a tertiary care hospital.
Data on patients with invasive Streptococcus anginosus group (SAG) infections is limited, as it's been considered commensal bacteria in the human microbiota. We conducted an analysis of SAG infections to assist clinicians in understanding their burden and clinical outcomes. A retrospective study of medical records, identifying invasive SAG bacteria of sterile-site isolates that were managed from May 2015 to April 2017, at a tertiary care hospital in Riyadh, Saudi Arabia. Demographic data, clinical presentation, site of infection, antibiotic use, and outcome were recorded and analyzed to identify factors associated with poor outcome and/or polymicrobial growth. We identified 105 cases of SAG infections in adults, with 52% of the patients being male and the mean age of 52.4 years with comorbidities occurring in more than half of the cases such as diabetes (38%) and malignancy (15%). Overall mortality was 6%, and it was statistically associated with age older than 65 years, polymicrobial growth and a history of malignancy. The infection frequencies were skin and soft tissue infections (SSTI; 55%), intra-abdominal infections (24%), bacteremia (14%), genitourinary infections (8.5%), and pleuropulmonary infections (5%). Abscesses accounted for 68% of cases. Polymicrobial infection (46%) with Enterobacteriaceae and Gram-negative anaerobes coincided with SAG infection. Polymicrobial growth was significantly associated with abscess formation, intra-abdominal source of infections, and poor outcome. In addition, death in patients with SAG was statistically associated with patients older than 65 years of age and those with history of cancer or transplant. SSTIs and intra-abdominal infections are the most common clinical presentations in our cohort. Bacteremia was uncommon; however, the prognosis is less favorable. Overall susceptibility to penicillin was 91%, therefore β-lactam antibiotics are the drug of choice and additional coverage for anaerobic and gram-negative bacteria should be considered for intra-abdominal collection and solid or organ abscesses.