CKD following kidney transplantation in children and adolescents.
There is under-recognition of comorbid conditions associated with chronic kidney disease (CKD) in children and adolescents after successful renal transplantation. Retrospective cross-sectional. Children and adolescents aged 1 to 20 years with kidney disease in a transplant (n = 45) and native-kidney-disease cohort (n = 102) matched for CKD stages. CKD stages were assigned using glomerular filtration rate measured by means of nuclear medicine studies. A single pediatric nephrology group cared for all patients. History of kidney transplantation. Complications of CKD (anemia, hypertension, acidosis, and bone mineral metabolism). The transplant (38% CKD stages 1 to 2, 62% CKD stages 3 to 5) and native-kidney (55% CKD stages 1 to 2, 45% CKD stages 3 to 5) cohorts were similar in demographic and baseline profiles; 68% of transplant recipients had 2 or more complications compared with 29% of native-kidney patients. After adjusting for baseline variables, the odds of having anemia was greater in transplant recipients (odds ratio, 9.7; 95% confidence interval, 3.9 to 24.6) at all CKD stages. The odds of having hypertension was particularly greater (odds ratio, 12.9; 95% confidence interval, 3.4 to 49.4) in transplant recipients with stages 1 to 2 CKD. No significant differences in bone mineral metabolism or acidosis were seen between groups. Retrospective cross-sectional design limits availability of data; lack of consistent protocols introduces treatment bias among physicians. Children with CKD after transplantation appear to have greater odds of having anemia and hypertension than those with CKD in native kidneys. We suggest that increased awareness and attention to these 2 modifiable risk factors for CKD and cardiovascular disease may improve outcomes after transplantation.


Rational construction of eukaryotic OFF-riboswitches that downregulate internal ribosome entry site-mediated translation in response to their ligands.
A strategy for rationally constructing a novel type of eukaryotic OFF-riboswitch, which ligand-dependently turns off translation mediated by an internal ribosome entry site (IRES), has been established. The theophylline-dependent IRES-based OFF-riboswitch obtained through the proposed strategy functioned well in wheat germ extract, independently from the downstream gene, indicating that it can regulate any gene. Despite the fact that it has one theophylline aptamer, its switching efficiency was as high as that of a previously reported theophylline-dependent OFF-riboswitch that was constructed by inserting three continuous theophylline aptamers into a 5' untranslated region in mRNA to downregulate the normal 5'-terminus-mediated translation. In addition, because the riboswitch part that was optimized in the theophylline-dependent IRES-based OFF-riboswitch, except for the aptamer domain, can be used as-is for other aptamer-ligand pairs, an arbitrary ligand-dependent IRES-based OFF-riboswitch is easy to construct with the corresponding well-minimized aptamer.


Biased Molecular Simulations for Free-Energy Mapping: A Comparison on the KcsA Channel as a Test Case.
The calculation of free-energy landscapes in proteins is a challenge for modern numerical simulations. As to the case of potassium ion channels is concerned, it is particularly interesting because of the nanometric dimensions of the selectivity filter, where the complex electrostatics is highly relevant. The present study aims at comparing three different techniques used to bias molecular dynamics simulations, namely Umbrella Sampling, Steered Molecular Dynamics, and Metadynamics, never applied all together in the past to the same channel protein. Our test case is represented by potassium ions permeating the selectivity filter of the KcsA channel.


The first molecular level monitoring of carbohydrate conversion to 5-hydroxymethylfurfural in ionic liquids. B2O3--an efficient dual-function metal-free promoter for environmentally benign applications.
The mechanistic nature of the conversion of carbohydrates to the sustainable platform chemical 5-hydroxymethylfurfural (5-HMF) was revealed at the molecular level. A detailed study of the key sugar units involved in the biomass conversion process has shown that the simple dissolution of fructose in the ionic liquid 1-butyl-3-methylimidazolium chloride significantly changes the anomeric composition and favors the formation of the open fructoketose form. A special NMR approach was developed for the determination of molecular structures and monitoring of chemical reactions directly in ionic liquids. The transformation of glucose to 5-HMF has been followed in situ through the detection of intermediate species. A new environmentally benign, easily available, metal-free promoter with a dual functionality (B(2)O(3)) was developed for carbohydrate conversion to 5-HMF.


Real world clinical feasibility of direct-from-specimen antimicrobial susceptibility testing of clinical specimens with unknown microbial load or susceptibility.
Within healthcare settings, physicians use antibiograms, which offer information on local susceptibility rates, as an aid in selecting empirical antibiotic therapy and avoiding the prescription of potentially ineffective drugs. While antibiograms display susceptibility and resistance data at hospital, city, or region-specific levels and ultimately enable the initiation of antibiogram-based empirical antibiotic treatment, AST reports at the individual patient level and guides treatments away from broad-spectrum antibiotics towards narrower-spectrum antibiotics or the removal of antibiotics entirely. Despite these advantages, AST traditionally requires a 48- to 72-h turn-around; this window of time can be critical for some antimicrobial therapeutic interventions. Herein, we present a direct-from-specimen AST to reduce the time between patient sampling and receipt of lab AST results. The biggest challenge of performing AST directly from unprocessed clinical specimens with an unknown microbial load is aligning the categorical susceptibility report with CLSI reference methods, which start from a fixed inoculum of 0.5 McFarland units prepared using colonies from a sub-culture. In this pilot clinical feasibility study using de-identified remnant specimens collected from MCW, we observed the high and low ends of microbial loads, demonstrating a final categorical agreement of 87.5% for ampicillin, 100% for ciprofloxacin, and 100% for sulfamethoxazole-trimethoprim.


Does it pay to develop a ground source heat pump system? Evidence from China.
The application potential and environmental benefits of ground source heat pump (GSHP) systems have become the focal points of decarbonization in the building sector. Synchronized and scientific analysis of GSHP systems' environmental and economic performance, however, remains lacking. This study analyzes the application prospects of GSHP systems via a life cycle assessment-based life cycle costing method, and considers China's actual status quo. The internal and external annual costs of a GSHP system per square meter are $ 4.05 and $ 1.37, respectively. Electricity generation and steel production are key processes to improve the environmental performance of a GSHP system further. Compared with coal-based heating, a GSHP system can mitigate 65%-95% of the environmental impact and 85% of external costs, except for the metal depletion impact which is 1.5 times higher than that of coal-based heating. In Shandong Province, promoting GSHP systems can substitute up to 69.4% of the district heating area, which implies reductions in fossil depletion, greenhouse gas emissions, human health impact, ecosystem quality impact, and external costs by up to 2.37 × 1010 kg oil eq, 1.08 × 1011 kg CO2 eq, 3.87 × 105 DALY, 1.18 × 103 Species. year, and $ 2.51 × 1010, respectively. In consideration of environmental and economic aspects, a GSHP system can exhibit benefits compared with coal-based heating after 2.34 years of operation. To improve the economic and environmental performance of GSHP systems, a series of recommendations on financial subsidies, renewable energy development, inter-regional power transmission, steel scrap utilization, and hydrogen reduction steelmaking is provided.


Characterization and expression analysis of gene encoding heme peroxidase HPX15 in major Indian malaria vector Anopheles stephensi (Diptera: Culicidae).
The interaction of mosquito immune system with Plasmodium is critical in determining the vector competence. Thus, blocking the crucial mosquito molecules that regulate parasite development might be effective in controlling the disease transmission. In this study, we characterized a full-length AsHPX15 gene from the major Indian malaria vector Anopheles stephensi. This gene is true ortholog of Anopheles gambiae heme peroxidase AgHPX15 (AGAP013327), which modulates midgut immunity and regulates Plasmodium falciparum development. We found that AsHPX15 is highly induced in mosquito developmental stages and blood fed midguts. In addition, this is a lineage-specific gene that has identical features and 65-99% amino acids identity with other HPX15 genes present in eighteen worldwide-distributed anophelines. We discuss that the conserved HPX15 gene might serve as a common target to manipulate mosquito immunity and arresting Plasmodium development inside the vector host.


Nilotinib and donor lymphocyte infusion in the treatment of Philadelphia-positive acute lymphoblastic leukemia (Ph+ ALL) relapsing after allogeneic stem cell transplantation and resistant to imatinib.
Prognosis of patients with Philadelphia-positive acute lymphoblastic leukemia (Ph+ ALL) relapsing after allogeneic stem cell transplantation (SCT) is dismal. Immunotherapy with donor lymphocyte infusion (DLI) and imatinib are rarely and/or transiently effective. Here we describe the case of a patient with imatinib-resistant post-transplant relapse of ALL, who received a combination of standard dose nilotinib and monthly DLI infusion. Therapy was well tolerated and the patient achieved and maintained a complete molecular remission. Our case provides a rationale for the combined use of a second line tyrosine kinase inhibitor and DLI in the treatment of relapsed Ph+ ALL.


Inhibitory effect of cyclo-oxygenase-2 inhibitor on the production of matrix metalloproteinases in rheumatoid fibroblast-like synoviocytes.
Cyclo-oxygenase (COX)-2 has been associated with inflammation in rheumatoid arthritis (RA), but its role in joint destruction remains unclear. In this study, we investigated the effect on cultured rheumatoid fibroblast-like synoviocytes (FLS) of the selective COX-2 inhibitor celecoxib on the expression of matrix metalloproteinases (MMPs), which play an important role in tissue degradation and angiogenesis in rheumatoid synovium. Treatment with nontoxic doses of celecoxib resulted in dose-dependent inhibition of MMP-1, -2, and -3 secretion from FLS when measured by enzyme-linked immunosorbent assay. Celecoxib suppressed proinflammatory cytokines (tumor necrosis factor-alpha and interleukin-1beta) induced augmentation of the gelatinolytic activity on zymography. These results suggest that COX-2 inhibitors might influence matrix degradation or angiogenesis in RA by downregulating the expression of various MMPs in rheumatoid FLS.


Predictors of treatment failure after top-hat procedure in squamous intraepithelial lesion.
The value of top-hat procedure during loop electrosurgical excision procedure (LEEP) for squamous intraepithelial lesions had remained controversial. This study aimed to evaluate whether top-hat specimens positive for cervical intraepithelial neoplasia (CIN) on histopathology can serve as an independent risk factor to predict treatment failure. We reviewed the medical records of patients who underwent LEEP and top-hat procedures in Peking University First Hospital between 2011 and 2016 and collected their follow-up data until January 2019. We compared the pathological risk factor of treatment failure. Multivariate analysis was carried out to clarify the independent determinant of treatment failure. A Cox model was used to assess the influence of different variables on cumulative treatment failure rates. This study included 295 cases for short-term treatment failure, and among them, 178 cases were used to study the long-term. The presence of CIN in top-hat was relevant to short-term treatment failure (OR = 9.64, 95% CI 2.55-36.4) despite a clear margin. On multivariate analysis, top-hat result (OR = 3.58, 95% CI 1.30-9.89), age ≥ 50 (OR = 10.2, 95%CI 3.64-28.3) and post-treatment HPV 16/18 infection (OR = 2.35, 95%CI 1.19-4.63) were independent risk factors in predicting short-term failure. In the Cox model, these factors were also associated with higher cumulative failure rates. The current study supported the predictive value of top-hat procedure in short-term failure after LEEP. Typically, women with positive top-hat need closer follow-up despite their negative margin status. Older women with positive top-hat findings and HPV 16/18 infections after the treatment suffer a higher risk of short-term failure.


Fusion and fission: membrane trafficking in animal cytokinesis.
Cytokinesis is the physical act of separating daughter cells, allowing them to become separate entities. Recent studies have revealed that membrane insertion for furrowing and scission of the residual bridge is a key aspect of animal cytokinesis.


Serum antibodies to surface proteins of Chlamydia trachomatis as candidate biomarkers of disease: results from the Baltimore Chlamydia Adolescent/Young Adult Reproductive Management (CHARM) cohort.
We previously observed that the nine-member family of autotransported polymorphic membrane proteins (Pmps) of Chlamydia trachomatis is variably expressed in cell culture. Additionally, C. trachomatis-infected patients display variable Pmp-specific serum antibody profiles indirectly suggesting expression of unique Pmp profiles is an adaptive response to host-specific stimuli during infection. Here, we propose that the host response to Pmps and other outer surface proteins may correlate with disease severity. This study tests this hypothesis using an ELISA that measures serum IgG antibodies specific for the nine C. trachomatis Pmp subtypes and four immunodominant antigens (MOMP, OmcB, Hsp60, ClpP) in 265 participants of the Chlamydia Adolescent/Young Adult Reproductive Management (CHARM) cohort. More C. trachomatis-infected females displayed high Pmp-specific antibody levels (cut-off Indexes) than males (35.9%-40.7% of females vs. 24.2%-30.0% of males), with statistical significance for PmpC, F and H (P < 0.05). Differences in Pmp-specific antibody profiles were not observed between C. trachomatis-infected females with a clinical diagnosis of pelvic inflammatory disease (PID) and those without. However, a statistically significant association between high levels of OmcB-specific antibody and a PID diagnosis (P< 0.05) was observed. Using antibody levels as an indirect measure of antigen expression, our results suggest that gender- and/or site-specific (cervix in females vs. urethra in males) stimuli may control pmp expression in infected patients. They also support the possible existence of immune biomarkers of chlamydial infection associated with disease and underline the need for high resolution screening in human serum.


Molecular identification of a recent type of canine distemper virus in Japan by restriction fragment length polymorphism.
Restriction fragment length polymorphism analysis was used to differentiate recent field viruses of canine distemper virus (CDU) from vaccine strains. Virus genomes were amplified by using reverse transcriptase-polymerase chain reaction in part of the haemagglutinin gene. After digestion with EcoRV, the PCR products of recent field isolates were cut into two fragments that differ from the uncut form of old strains including all of vaccine strains. This method could be applied to fresh or stored brains, spleens and peripheral blood mononuclear cells of infected dogs. This molecular approach is useful for determining the causative agent of postvaccinated CDV infection.


The essential role of acetyllysine binding by the YEATS domain in transcriptional regulation.
The YEATS domains of AF9 and Taf14 have recently been found to recognize the histone H3K9ac modification. In this commentary, we discuss the mechanistic and biological implications of this interaction. We compare structures of the YEATS-H3K9ac complexes the highlighting a novel mechanism for the acetyllysine recognition through the aromatic cage. We also summarize the latest findings underscoring a critical role of the acetyllysine binding function of AF9 and Taf14 in transcriptional regulation and DNA repair.


The Importance of Employee Breaks.
The occupational and environmental health nurse should encourage workers to take work breaks. Breaks enable all workers to perform at their best.


The process of reinnervation in the dentate gyrus of adult rats: an ultrastructural study of changes in presynaptic terminals as a result of sprouting.
The present study was undertaken to define the ultrastructure of synapses of the crossed temporodentate pathway after they had sprouted to reinnervate the dentate gyrus following the destruction of the normal ipsilateral temporodentate pathway. The synapses of the sprouted crossed temporodentate pathway were identified at the EM level by using autoradiographic techniques and by evaluating the degeneration of the pathway following secondary lesions. Both EM autoradiography and EM degeneration revealed that the terminals of the sprouted crossed temporodentate pathway formed asymmetric synapses on spines; individual terminals appeared to make more synaptic contacts per terminal (multiple synapses) than in the case of the normal crossed pathway. In the two lesioned animals exhibiting the best labeling, labeled terminals made an average of 3.0 +/- 2.2 and 2.0 +/- 1.3 contacts per terminal. In contrast, labeled terminals in normal animals exhibited only one contact per terminal. The terminals of the sprouted pathway were also larger than those of the normal crossed pathway. The synapses of the crossed temporodentate pathway that degenerated after a secondary lesion of the entorhinal cortex exhibited both electron-lucent and electron-dense forms of degeneration at 2 days postlesion. In two animals that were quantitatively analyzed, the density of degenerating synaptic terminals was 281 and 218/10,000 microns2 in the terminal field of the sprouted crossed pathway. These values are much higher than in normal animals, where the density of degenerating synaptic terminals was only 2.12/10,000 microns2 at 2 days postlesion.(ABSTRACT TRUNCATED AT 250 WORDS)


Collagen metabolic disorder induced by oxidative stress in human uterosacral ligament‑derived fibroblasts: A possible pathophysiological mechanism in pelvic organ prolapse.
Pelvic organ prolapse (POP) is a global health problem, for which the pathophysiological mechanism remains to be fully elucidated. The loss of extracellular matrix protein has been considered to be the most important molecular basis facilitating the development of POP. Oxidative stress (OS) is a well‑recognized mechanism involved in fiber metabolic disorders. The present study aimed to clarify whether OS exists in the uterosacral ligament (USL) with POP, and to investigate the precise role of OS in collagen metabolism in human USL fibroblasts (hUSLFs). In the present study, 8‑hydroxyguanosine (8‑OHdG) and 4 hydroxynonenal (4‑HNE), as oxidative biomarkers, were examined by immunohistochemistry to evaluate oxidative injury in USL sections in POP (n=20) and non‑POP (n=20) groups. The primary cultured hUSLFs were treated with exogenous H2O2 to establish an original OS cell model, in which the expression levels of collagen, type 1, α1 (COL1A1), matrix metalloproteinase (MMP)‑2, tissue inhibitor of metalloproteinase (TIMP)‑2 and transforming growth factor (TGF)‑β1 were evaluated by western blot and reverse transcription‑quantitative polymerase chain reaction analyses. The results showed that the expression levels of 8‑OHdG and 4‑HNE in the POP group were significantly higher, compared with those in the control group. Collagen metabolism was regulated by H2O2 exposure in a concentration‑dependent manner, in which lower concentrations of H2O2 (0.1‑0.2 mM) stimulated the anabolism of COL1A1, whereas a higher concentration (0.4 mM) promoted catabolism. The expression levels of MMP‑2, TIMP‑2 and TGF‑β1 exhibited corresponding changes with the OS levels. These results suggested that OS may be involved in the pathophysiology of POP by contributing to collagen metabolic disorder in a severity‑dependent manner in hUSLFs, possibly through the regulation of MMPs, TIMPs and TGF‑β1 indirectly.


An Unexpected Transient Breakdown of the Blood Brain Barrier Triggers Passage of Large Intravenously Administered Nanoparticles.
The highly restrictive blood-brain barrier (BBB) plays a critically important role in maintaining brain homeostasis and is pivotal for proper neuronal function. The BBB is currently considered the main limiting factor restricting the passage of large (up to 200 nm) intravenously administered nanoparticles to the brain. Breakdown of the barrier occurs as a consequence of cerebrovascular diseases and traumatic brain injury. In this article, we report that remote injuries in the CNS are also associated with BBB dysfunction. In particular, we show that a focal partial transection of the optic nerve triggers a previously unknown transient opening of the mammalian BBB that occurs in the visual centres. Importantly, we demonstrate that this transient BBB breakdown results in a dramatic change in the biodistribution of intravenously administered large polymeric nanoparticles which were previously deemed as BBB-impermeable.


Attenuation of Proximal Junctional Kyphosis Using Sublaminar Polyester Tension Bands: A Biomechanical Study.
To investigate the effect of sublaminar polyester tension bands on the biomechanics of the motion segments proximal to a long fusion construct. This was a human cadaveric biomechanical study. Pure moments of 4 Nm and 8 Nm were applied to the native spine and the instrumented spine, respectively (n = 8). The test conditions included native spine (T7-L2), fused (T10-L2), fused + bilateral tethers tensioned to 250 N at T9-T10 (tethers 250 N), fused + tethers tensioned to 350 N (tethers 350 N), fused (T11-L2) + tethers tensioned to 250 N at T9-T10 and 350 N at T10-T11 (2-level tethers), fused (T10-L2) + hand-tied suture loop through the spinous processes at T9-T10 (suture loop), and fused (T10-L2) with the T9-T10 interspinous and supraspinous ligaments cut (cut ISL/SSL). The flexion range of motion (ROM) at T9-T10 of the fused spine, loaded at 8 Nm, increased to 162% of the native spine loaded at 4 Nm. The average flexion ROM at T9-T10 for tethers 250 N, tethers 350 N, 2-level tethers, suture loop, and cut ISL/SSL were 85% (P < 0.0001), 70% (P < 0.0001), 93% (P < 0.0001), 141% (P = 0.13), and 177% (P = 0.66) of the native spine at 4 Nm, respectively (P values vs. fused). Sublaminar polyester bands can modulate the biomechanical flexion ROM as a function of the band pretension and provide a more consistent and tunable technique than hand-tying a suture loop between the spinous processes.


Protective effects of topiramate on acetic acid-induced colitis in rats through the inhibition of oxidative stress.
Ulcerative colitis is an intestinal inflammatory condition characterized by a rise in inflammatory mediator production and oxidative stress. Topiramate is an anticonvulsant agent with effectiveness on a wide range of seizures, which is anti-oxidative. This study aims to examine the protective effects of topiramate on acetic acid-induced ulcerative colitis in rats. Rats were randomly divided into four groups as follows: control, acetic acid, acetic acid + topiramate, and acetic acid + dexamethasone groups. Topiramate (100 mg/kg/day) or dexamethasone (2 mg/kg/day) was administered for six consecutive days, and ulcerative colitis was induced on the first day of the study by transrectal administration of 4% acetic acid. Four hours after the last dose of treatments, animals of each group were sacrificed, and colon tissues were removed for further macroscopic, histopathologic, and biochemical analyses. Treatment with topiramate markedly decreased colonic lesions and macroscopic scores as well as the improvement of histopathologic changes. Topiramate also effectively decreased the levels of malondialdehyde and upregulated the activity of anti-oxidative enzymes, including catalase, superoxide dismutase, and glutathione peroxidase. Our results reveal that the administration of topiramate ameliorates acetic acid-induced colitis in rats via anti-oxidative properties, and further studies may introduce it as an effective therapeutic candidate to decrease ulcerative colitis severity.


Robust regularity in γ-soft nuclei and its microscopic realization.
γ softness in atomic nuclei is investigated in the framework of energy density functionals. By mapping constrained microscopic energy surfaces for a set of representative nonaxial medium-heavy and heavy nuclei to a Hamiltonian of the proton-neutron interacting boson model (IBM-2) containing up to three-body interactions, low-lying collective spectra and transition rates are calculated. Observables are analyzed that distinguish between the two limiting geometrical pictures of nonaxial nuclei: the rigid-triaxial rotor and the γ-unstable rotor. It is shown that neither of these pictures is realized in actual nuclei, and that a microscopic description leads to results that are almost exactly in between the two geometrical limits. This finding points to the optimal choice of the IBM Hamiltonian for γ-soft nuclei.


Glyceraldehyde-3-phosphate dehydrogenase, a glycolytic enzyme present in the periplasm of Aeromonas hydrophila.
This is the first report describing the glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), as a protein associated with the cell envelope of a gram-negative bacterium (Aeromonas hydrophila). Dose-dependent GAPDH activity was detected in whole bacterial cells from exponentially growing cultures, indicating that an active form of GAPDH is located outside the plasma membrane. This activity represents roughly 10-20% of total cell activity, and it is not reduced by pretreatment of the cells with trypsin. Assays with soluble GAPDH indicate that the activity measured in intact cells does not originate by rebinding to intact cells of cytosolic enzyme released following cell lysis. GAPDH activity levels detected in intact cells varied during the growth phase. The relationship between GAPDH activity and cell culture density was not linear, showing this activity as a major peak in the late-logarithmic phase (A600 = 1.1-1.3), and a decrease when cells entered the stationary phase. The late exponential growing cells showed a GAPDH activity 3 to 4-fold higher than early growing or stationary cells. No activity was detected in culture supernatants. Enzymatic and Western-immunoblotting analysis of subcellular fractions (cytosol, whole and outer membranes, and periplasm) showed that GAPDH is located in the cytosol, as expected, and also in the periplasm. These results place the periplasmic GAPDH of A. hydrophila into the family of multifunctional microbial cell wall-associated GAPDHs which retain their catalytic activity.


GluN2B S1303 phosphorylation by CaMKII or DAPK1: no indication for involvement in ischemia or LTP.
Binding of two different CaM kinases, CaMKII and DAPK1, to the NMDA-type glutamate receptor (NMDAR) subunit GluN2B near S1303 has been implicated in excitotoxic/ischemic neuronal cell death. The GluN2BΔCaMKII mutation (L1298A, R1300Q) is neuroprotective but abolishes only CaMKII but not DAPK1 binding. However, both kinases can additionally phosphorylate GluN2B S1303. Thus, we here tested S1303 phosphorylation for possible contribution to neuronal cell death. The GluN2BΔCaMKII mutation completely abolished phosphorylation by CaMKII and DAPK1, suggesting that the mutation could mediate neuroprotection by disrupting phosphorylation. However, S1303 phosphorylation was not increased by excitotoxic insults in hippocampal slices or by global cerebral ischemia induced by cardiac arrest and cardiopulmonary resuscitation in vivo. In hippocampal cultures, S1303 phosphorylation was induced by chemical LTD but not LTP stimuli. These results indicate that the additional effect of the GluN2BΔCaMKII mutation on phosphorylation needs to be considered only in LTD but not in LTP or ischemia/excitotoxicity.


Single Versus Dual Lead Atrioventricular Sequential Pacing for Acquired Atrioventricular Block During Transcatheter Aortic Valve Implantation Procedures.
Atrioventricular block (AVB) after transcatheter aortic valve implantation (TAVI) is common. Pacing platforms that preserve atrioventricular (AV) synchrony may be beneficial in these patients. Single lead AV sequential pacemakers (VDD) allow AV synchrony and, by virtue of implanting a single lead, can be advantageous when compared with traditional dual lead (DDD) pacemakers. The objective of this retrospective case-controlled study was to compare the strategy of AV sequential pacing with a VDD versus DDD pacemaker in consecutive TAVI patients (n = 120) with acquired AVB. Patients were classified as receiving a VDD (n = 73) or DDD (n = 47) pacemaker. Procedure characteristics, complications, and the need for long-term ventricular pacing reported. Pacemaker implantation time (51 vs 66 minutes and p <0.001), fluoroscopy time (3.7 vs 7.8 minutes and p = 0.004), and radiation exposure (dose area product: 6.5 vs 15.3 mGy/cm2 and p = 0.006) were lower in patients receiving VDD pacemakers. Procedural complications were similar in the 2 groups. Overall, 59% of the cohort required ≥50% ventricular pacing at 1-year follow-up. In conclusion, VDD pacemaker implantation was associated with shorter procedure times, lower radiation exposure, and similar safety outcomes when compared with DDD pacemaker implantation in TAVI patients with acquired AVB.


Synthetic azo dye bio-decolorization by Priestia sp. RA1: process optimization and phytotoxicity assessment.
Azo compounds represent the most diverse group of colorants widely employed in industrial sectors. Being highly toxic and recalcitrant compound, azo dyes pose a threat to plants, animals, and humans. In the present report, bio-decolorization of azo dye, reactive black 5, was evaluated by newly isolated Priestia sp. RA1. Strain RA1 was able to decolorize 97% of 100 ppm reactive black 5 in 60 h. Specific activity of dye decolorization was found to be 0.233 μmol min-1 g-1 dry cells. Successful decolorization over a broad range of pH, salinity, temperature, and initial dye concentration was observed. Phytotoxicity assay on agriculturally important crops showed considerable difference in percentage seed germination and growth when treated with original and bio-decolorized dye samples. Bio-decolorization at high dye concentrations, promising decolorization rate, and non-toxic nature of treated products suggest the potential of strain RA1 for bioremediation of dye-contaminated water and its re-use in the industries.


Application of QMRA to MAR operations for safe agricultural water reuses in coastal areas.
A pathogenic Escherichia coli (E.coli) O157:H7 and O26:H11 dose-response model was set up for a quantitative microbial risk assessment (QMRA) of the waterborne diseases associated with managed aquifer recharge (MAR) practices in semiarid regions. The MAR facility at Forcatella (Southern Italy) was selected for the QMRA application. The target counts of pathogens incidentally exposed to hosts by eating contaminated raw crops or while bathing at beaches of the coastal area were determined by applying the Monte Carlo Markov Chain (MCMC) Bayesian method to the water sampling results. The MCMC provided the most probable pathogen count reaching the target and allowed for the minimization of the number of water samplings, and hence, reducing the associated costs. The sampling stations along the coast were positioned based on the results of a groundwater flow and pathogen transport model, which highlighted the preferential flow pathways of the transported E. coli in the fractured coastal aquifer. QMRA indicated tolerable (<10-6 DALY) health risks for bathing at beaches and irrigation with wastewater, with 0.4 infectious diseases per year (11.4% probability of occurrence) associated with the reuse of reclaimed water via soil irrigation even though exceeding the E. coli regulation limit of 10 CFU/100 mL by five times. The results show negligible health risk and insignificant impacts on the coastal water quality due to pathogenic E. coli in the wastewater used for MAR. However, droughts and reclaimed water quality can be considered the main issues of MAR practices in semiarid regions suggesting additional reclaimed water treatments and further stress-tests via QMRAs by considering more persistent pathogens than E. coli.


Novel MNX1 mutations and genotype-phenotype analysis of patients with Currarino syndrome.
Currarino syndrome (CS) is a specific complex of congenital caudal anomalies, including anorectal malformations, presacral mass and sacral anomalies. Mutations in the MNX1 gene are closely related to CS and occur in almost all familial cases and less than half of sporadic patients. We investigated the spectrum of MNX1 pathogenic variants and associated clinical features in Chinese patients with CS. Seventeen index patients from 16 families were recruited from 2015 to 2018. All patients were diagnosed with CS and treated at the Capital Institute of Pediatrics Affiliated Children's Hospital. Genetic testing was applied to identify mutations in CS patients and their relatives by whole-exome sequencing and Sanger sequencing. Functional verification was performed for a recurrent noncanonical splice site variant in MNX1 with a minigene splicing assay. In 17 CS patients, 14 were complete CS and 3 were mild CS. Nine variants in MNX1 were identified in 11 patients, and these included two frameshift mutations (p.Leu223Leufs*61, p.X402Serfs*70), four nonsense mutations (p.Gly42X, p.Cys88X, p.Gln24X, p.Cys241X), one missense mutation (p.Trp288Leu), one splice region variant (c.691 + 3G > T) and one polyalanine polymorphism (p.Ala135insAlaAla). Seven of these nine variants have never been reported. Pathogenic MNX1 mutations were found in 100% (4/4) of familial and 46% (6/13) of sporadic patients. Our study expanded the mutation spectrum of MNX1 and provided clinical and genetic analyses of seventeen CS patients from mainland China.


Efficacy of an artificial neural network-based approach to endoscopic ultrasound elastography in diagnosis of focal pancreatic masses.
By using strain assessment, real-time endoscopic ultrasound (EUS) elastography provides additional information about a lesion's characteristics in the pancreas. We assessed the accuracy of real-time EUS elastography in focal pancreatic lesions using computer-aided diagnosis by artificial neural network analysis. We performed a prospective, blinded, multicentric study at of 258 patients (774 recordings from EUS elastography) who were diagnosed with chronic pancreatitis (n = 47) or pancreatic adenocarcinoma (n = 211) from 13 tertiary academic medical centers in Europe (the European EUS Elastography Multicentric Study Group). We used postprocessing software analysis to compute individual frames of elastography movies recorded by retrieving hue histogram data from a dynamic sequence of EUS elastography into a numeric matrix. The data then were analyzed in an extended neural network analysis, to automatically differentiate benign from malignant patterns. The neural computing approach had 91.14% training accuracy (95% confidence interval [CI], 89.87%-92.42%) and 84.27% testing accuracy (95% CI, 83.09%-85.44%). These results were obtained using the 10-fold cross-validation technique. The statistical analysis of the classification process showed a sensitivity of 87.59%, a specificity of 82.94%, a positive predictive value of 96.25%, and a negative predictive value of 57.22%. Moreover, the corresponding area under the receiver operating characteristic curve was 0.94 (95% CI, 0.91%-0.97%), which was significantly higher than the values obtained by simple mean hue histogram analysis, for which the area under the receiver operating characteristic was 0.85. Use of the artificial intelligence methodology via artificial neural networks supports the medical decision process, providing fast and accurate diagnoses.


Binding of pyridine derivatives to cytochrome P-450.
The spectral binding constants, Ks, of pyridine, methylpyridine, and dimethylpyridines were determined. The Ks values of the compounds were discussed in terms of mechanisms of possible bonding to cytochrome P-450.


Mobilization of vitamin B12 transporters alters competitive dynamics in a human gut microbe.
The functional and genomic diversity of the human gut microbiome is shaped by horizontal transfer of mobile genetic elements (MGEs). Characterized MGEs can encode genes beneficial for their host's self-defense (e.g., antibiotic resistance) or ability to compete for essential or limited resources (e.g., vitamins). Vitamin B12 and related compounds (corrinoids) are critical nutrients that enable colonization by members of the common gut microbe phylum, the Bacteroidetes. Herein, we identify a distinct class of MGEs in the Bacteroidetes responsible for the mobilization and exchange of the genes required for transport of corrinoids, a group of cyclic tetrapyrrole cofactors including vitamin B12 (btuGBFCD). This class includes two distinct groups of conjugative transposons (CTns) and one group of phage. Conjugative transfer and vitamin B12 transport activity of two of the CTns were confirmed in vitro and in vivo, demonstrating the important role MGEs play in distribution of corrinoid transporters in the Bacteroidetes.


Picoplankton diversity in an oligotrophic and high salinity environment in the central Adriatic Sea.
By combining qualitative 16S metabarcoding and quantitative CARD-FISH methods with neural gas analysis, different patterns of the picoplankton community were revealed at finer taxonomic levels in response to changing environmental conditions in the Adriatic Sea. We present the results of a one-year study carried out in an oligotrophic environment where increased salinity was recently observed. We have shown that the initial state of community structure changes according to environmental conditions and is expressed as qualitative and quantitative changes. A general pattern of increasing diversity under harsh environmental conditions, particularly under the influence of increasing salinity at the expense of community abundance was observed. Considering the trend of changing seawater characteristics due to climate change, this study helps in understanding a possible structural change in the microbial community of the Adriatic Sea that could affect higher levels of the marine food web.


Crystal structure of the kinase domain of serum and glucocorticoid-regulated kinase 1 in complex with AMP PNP.
Serum and glucocorticoid-regulated kinase 1 (SGK1) is a serine/threonine protein kinase of the AGC family which participates in the control of epithelial ion transport and is implicated in proliferation and apoptosis. We report here the 1.9 A crystal structure of the catalytic domain of inactive human SGK1 in complex with AMP-PNP. SGK1 exists as a dimer formed by two intermolecular disulfide bonds between Cys258 in the activation loop and Cys193. Although most of the SGK1 structure closely resembles the common protein kinase fold, the structure around the active site is unique when compared to most protein kinases. The alphaC helix is not present in this inactive form of SGK1 crystal structure; instead, the segment corresponding to the C helix forms a beta-strand that is stabilized by the N-terminal segment of the activation loop through a short antiparallel beta-sheet. Since the differences from other kinases occur around the ATP binding site, this structure can provide valuable insight into the design of selective and highly potent ATP-competitive inhibitors of SGK1 kinase.


A sunlight-induced rapid synthesis of silver nanoparticles using sodium salt of N-cholyl amino acids and its antimicrobial applications.
Aqueous solution containing two additives, silver nitrate (AgNO(3)) and sodium salt of N-cholyl amino acid were irradiated by sunlight for the synthesis of spherical shaped AgNPs without the need for an additional stabilizer or capping agent. Variations of N-cholyl amino acid concentration provided good control over the morphology of the AgNPs, while the carboxylate group of bile salt reduced the Ag(+) ions and the amide group binds strongly to the surface of the NPs. The optical properties, morphology of the AgNPs were characterized by UV-visible, transmission electron microscopy (TEM) and dynamic light scattering (DLS) studies. The interaction of N-cholyl amino acid on the AgNPs surface was studied using cyclic voltammetry and FT-IR techniques. The reduction process was completed within 5 min and the synthesized AgNPs were stable for more than 6 months. The possible mechanism of N-cholyl amino acid on the reduction and stabilization of AgNPs is also discussed. The antimicrobial activity of N-cholyl amino acid capped AgNPs against Escherichia coli, Staphylococcus aureus and Pseudomonas aeroginosa using Mueller Hinton broth and the antifungal activity against Candida albicans, Candida krusei and Candida tropicalis using RPMI broth were determined by MIC studies as per CLSI guidelines.


Differential distribution of acetylcholinesterase activity among vasopressin- and oxytocin-containing supraoptic magnocellular neurons.
Acetylcholinesterase activity was demonstrated histochemically at light- and electron-microscopic levels, in Vibratome sections of the supraoptic nucleus of fixed hypothalami derived from osmotically stimulated and unstimulated Long Evans rats, from homozygous Brattleboro rats with hypothalamic diabetes insipidus, from lactating rats, from normal adult male house mice (Mus musculus) and from mice with hereditary nephrogenic diabetes insipidus (di/di). Reaction product was located in supraoptic magnocellular neurons; in dorsal and rostral aspects of the supraoptic nuclei lightly stained cells predominate, whereas in ventral and caudal regions densely staining perikarya predominate. Pre- and post-embedding immunocytochemical detection of oxytocin-neurophysin or vasopressin-neurophysin, combined with acetylcholinesterase histochemistry, showed that the lightly staining cells are oxytocinergic, and the densely staining cells vasopressinergic. Osmotic stimulation of the animals, either by substitution of drinking water for 3 days with 2.5% saline or reason of genetic defects which result in diabetes insipidus, enhanced the acetylcholinesterase activity of the vasopressin neurons but had little effect on the weekly acetylcholinesterase-reactive oxytocin cells. Acetylcholinesterase activity was particularly marked in the hypertrophied abnormal magnocellular neurons of homozygous Brattleboro rats which do not release significant amounts of vasopressin. The increased acetylcholinesterase activity in osmotically stimulated animals cannot, therefore, be a function of vasopressin. Acetylcholinesterase activity was also detected in large multipolar neurons lying dorsolateral to the supraoptic nucleus, and in their fine axonal processes which project towards the supraoptic nucleus. A very few synaptic boutons surrounded by acetylcholinesterase reaction product were found in contact with magnocellular neuron basal dendrites. However, much of the punctate acetylcholinesterase reactivity observed at the light microscopic level and previously interpreted as representing the loci of cholinergic synaptic boutons was shown to be intracellular, and probably caused by acetylcholinesterase activity in some large, secondary lysosomes.


Arsenic induces pancreatic dysfunction and ferroptosis via mitochondrial ROS-autophagy-lysosomal pathway.
Chronic arsenic exposure is a significantly risk factor for pancreatic dysfunction and type 2 diabetes (T2D). Ferroptosis is a newly identified iron-dependent form of oxidative cell death that relies on lipid peroxidation. Previous data have indicated that ferroptosis is involved in various diseases, including cancers, neurodegenerative diseases, and T2D. However, the concrete effect and mechanism of ferroptosis on pancreatic dysfunction triggered by arsenic remains unknown. In this study, we verified that ferroptosis occurred in animal models of arsenic-induced pancreatic dysfunction through assessing proferroptotic markers and morphological changes in mitochondria. In vitro, arsenic caused execution of ferroptosis in a dose-dependent manner, which could be significantly reduced by ferrostatin-1. Additionally, arsenic damaged mitochondria manifested as diminishing of mitochondrial membrane potential, reduced cytochrome c level and production of mitochondrial reactive oxygen species (MtROS) in MIN6 cells. Using the Mito-TEMPO, we found the autophagy level and subsequent ferroptotic cell death induced by arsenic were both alleviated. With autophagy inhibitor chloroquine, we further revealed that ferritin regulated ferroptosis through the MtROS-autophagy pathway. Collectively, NaAsO2-induced ferroptotic cell death is relied on the MtROS-dependent autophagy by regulating the iron homeostasis. Ferroptosis is involved in pancreatic dysfunction triggered by arsenic, and arsenic-induced ferroptosis involves MtROS, autophagy, ferritin.


A comparison of formalin and GEWF in fixation of colorectal carcinoma specimens: rates of lymph node retrieval and effect on TNM staging.
The Royal College of Pathologists recommend that a median of at least 12 lymph nodes should be harvested during pathological staging of colorectal cancer. It is not always easy to harvest the required number, especially in patients with rectal cancer receiving neoadjuvant therapy. Lymph node revealing solutions, for example, GEWF, may improve nodal yield. GEWF is safe, cheap and easy to use. In a controlled trial, lymph node yields were compared after secondary specimen dissection following either 24 h of further fixation in formalin (n=101) or GEWF immersion (n=99). The number, size and tumour status of additional lymph nodes identified were compared between groups. Twenty-seven cases that received long-course neoadjuvant therapy were also assessed. Median lymph node yield at primary dissection met national standards overall (19) but also in the long-course neoadjuvant therapy group (13). Lymph nodes were smaller in neoadjuvant cases compared with non-neoadjuvant cases (mean size range 1.3-5.6 mm vs 1.5-8.9 mm). The use of further fixation and GEWF detected more nodes at secondary dissection. The mean number of additional nodes harvested was greater with formalin (8.3) than GEWF (7.3). There was no significant difference in the mean size of the additional lymph nodes detected between groups (point estimate 1.02; 95% CI -0.58 to 2.63; p=0.211). Upstaging triggering adjunct chemotherapy occurred in 1% (2/200) of cases. The routine use of adjunct techniques to identify additional lymph nodes is unnecessary with underlying high-quality dissection practice. Emphasis should be placed upon education and training, spending appropriate time dissecting and ensuring specimens are sufficiently fixed beforehand.


Suture line recurrence in jejunal pouch replaced after total gastrectomy for gastric cancer.
We report a case of suture line recurrence in a jejunal pouch, diagnosed 4 months after total gastrectomy for advanced gastric cancer. The jejunal pouch was made with a linear stapler, without intraluminal irrigation being carried out before anastomosis, and was replaced in an interposition fashion. We propose that the recurrence was caused by the implantation of exfoliated cancer cells in either the intraluminal mucus or on a contaminated stapling device.


Suicide assessment and treatment in pediatric primary care settings.
This article will briefly review screening for depression and suicidal ideation in primary care and school-based clinics, with a focus on in-depth screening for imminent suicide risk, developing a safety plan, and incorporating handoffs to urgent and emergency mental health care personnel. The article will cover current definitions of levels of suicidal risk and clinic-based protocols for a team approach to adolescents in crisis. To provide primary care and behavioral health nurses with evidence-based suicide risk screening and assessment tools and best practices for using them in patient-centered encounters with adolescents with suicidal thinking or behavior. Journal articles, books, and reports. Past studies have shown that many individuals who died by suicide had seen a primary care provider in 30 days before their deaths. Nurses in primary care settings should develop clinic-based protocols for screening all adolescents for suicide risk, developing safety plans, and providing suicidal youth and families with monitoring, appropriate referrals, follow-up, and support.


Optimization of near-infrared fluorescent sentinel lymph node mapping for vulvar cancer.
Near-infrared fluorescence imaging has the potential to improve sentinel lymph node mapping in vulvar cancer, which was assessed in the current study. Furthermore, dose optimization of indocyanine green adsorbed to human serum albumin was performed. Nine vulvar cancer patients underwent the standard sentinel lymph node procedure using (99m)technetium-nancolloid and patent blue. In addition, intraoperative imaging was performed after peritumoral injection of 1.6 mL of 500, 750, or 1000 μM of indocyanine green adsorbed to human serum albumin. Near-infrared fluorescence sentinel lymph node mapping was successful in all patients. A total of 14 sentinel lymph nodes (average, 1.6; range, 1-4) were detected: 14 radioactive (100%), 11 blue (79%), and 14 near-infrared fluorescent (100%). This study demonstrates feasibility and accuracy of sentinel lymph node mapping using indocyanine green adsorbed to human serum albumin. Considering safety, cost, and pharmacy preferences, an indocyanine green adsorbed to human serum albumin concentration of 500 μM appears optimal for sentinel lymph node mapping in vulvar cancer.


Network architecture of non-coding RNAs provides insights into the pathogenesis of upper tract urothelial carcinoma.
Numerous studies suggested that non-coding RNA modifications play an important role in upper tract urothelial carcinoma (UTUC), but few have depicted the architecture of non-coding RNA on the pathological process of UTUC. We aimed to better understand the pathogenesis of UTUC and provide precision medicine references of non-coding RNA when managing UTUC patients. PubMed, Cochrane Library, Embase, and Scopus were searched for UTUC until December 31, 2020. Methodological quality assessment was conducted according to NIH recommendations. Enrichment analyses and network analyses were conducted to explore the interactions of miRNA with genes and other non-coding RNAs. Survival analyses were performed to validate the novel genes. A total of 12 pairs of UTUC tumors and adjacent normal tissues were also included to validate the gene expressions regulated by miRNAs from the miRNA-gene network. Thirteen studies with 945 patients were eligible, investigating 106 miRNAs mutations. The quality of all the studies was fair to good. Most miRNAs were enriched in tissue/organs, diseases, and specific anti-cancer drugs (false discovery rate <0.05). Other non-coding RNAs, i.e.,: miR-34a, DLGAP1-AS1, USP39, and RNA5SP479, were highlighted by network analyses to have potential in the pathogenesis of UTUC. Top hub genes in the miRNA-gene network, namely ZNF460, NUFIP2, and E2F3, were all validated by survival analysis(P < 0.05). Using own cohort data, the differential expression analyses identified 368 overlapped significant genes, including above 3 hub genes (false discovery rate <0.05). Novel biomarkers identified in our studies might play essential roles in UTUC, from the perspectives of the molecule, tissue/organ, diagnosis, treatment, and prognosis. Candidate biomarkers could be significant references for personalized and target therapies.


Complement membrane attack (MAC) in idiopathic IgA-glomerulonephritis.
Antigens of the membrane attack complex of complement (MAC), such as C5, C6, C9 and MAC-related neoantigen(s), were demonstrated in the mesangium of 23 cases with IgA-glomerulonephritis (IgA-GN) and two cases with Henoch-Schönlein purpura nephritis (HSP). High specificity of the polyclonal antibodies was verified by dot-blot analysis. Control specimens lacking immunoglobulin deposits were negative for MAC-related antigens. Markers of classical pathway activation (Clq and C4) were observed only in two of 24 and one of 23 cases of IgA-GN and HSP, respectively. Glomerular distribution patterns (mesangial vs. mesangio-peripheral) of immunoglobulin or complement deposits were correlated for IgA and C3b/iC3b (P less than 0.002), for IgA and properdin (P less than 0.002) and for IgA and MAC neoantigens (P less than 0.01). Double immunostaining experiments revealed co-localization of IgA and MAC neoantigens at identical mesangial and capillary sites. Glomerular distribution of the less pronounced IgG or IgM deposits did not correlate with that of any complement-derived antigen. The pattern of MAC-related antigens was found to be uniformly either mesangial or mesangio-peripheral. Staining for MAC-related antigens was less intense in IgA-GN cases with minimal glomerular lesions than in cases with more advanced non-sclerosing lesions. IgA, C3d, and MAC localized in corresponding glomerular sites. This is consistent with complete local activation of complement by glomerular IgA deposits via the alternative pathway. The possibility exists that MAC plays a pathogenetic role, such as by irritation of bystander cells, in IGA-GN and HSP.


Failure of computerized tomography to differentiate between radiation necrosis and cerebral tumour.
Post-radiation cerebral necrosis simulating tumours was found in 2 patients by computerized axial tomography (CAT). The appearance was indistinguishable from that of a recurrent tumour in one patient and a glioma in the second. The significance of the scan findings was only realized after surgery and histological examination. The investigation appears to offer no assistance in differentiating between these conditions.


On the role of the immunoproteasome in transplant rejection.
The immunoproteasome is expressed in cells of hematopoietic origin and is induced during inflammation by IFN-γ. Targeting the immunoproteasome with selective inhibitors has been shown to be therapeutically effective in pre-clinical models for autoimmune diseases, colitis-associated cancer formation, and transplantation. Immunoproteasome inhibition prevents activation and proliferation of lymphocytes, lowers MHC class I cell surface expression, reduces the expression of cytokines of activated immune cells, and curtails T helper 1 and 17 cell differentiation. This might explain the in vivo efficacy of immunoproteasome inhibition in different pre-clinical disease models for autoimmunity, cancer, and transplantation. In this review, we summarize the effect of immunoproteasome inhibition in different animal models for transplantation.


Dominantly inherited Alzheimer's disease in Latin America: Genetic heterogeneity and clinical phenotypes.
A growing number of dominantly inherited Alzheimer's disease (DIAD) cases have become known in Latin American (LatAm) in recent years. However, questions regarding mutation distribution and frequency by country remain open. A literature review was completed aimed to provide estimates for DIAD pathogenic variants in the LatAm population. The search strategies were established using a combination of standardized terms for DIAD and LatAm. Twenty-four DIAD pathogenic variants have been reported in LatAm countries. Our combined dataset included 3583 individuals at risk; countries with highest DIAD frequencies were Colombia (n = 1905), Puerto Rico (n = 672), and Mexico (n = 463), usually attributable to founder effects. We found relatively few reports with extensive documentation on biomarker profiles and disease progression. Future DIAD studies will be required in LatAm, albeit with a more systematic approach to include fluid biomarker and imaging studies. Regional efforts are under way to extend the DIAD observational studies and clinical trials to Latin America.


Transport of intensity diffraction tomography with non-interferometric synthetic aperture for three-dimensional label-free microscopy.
We present a new label-free three-dimensional (3D) microscopy technique, termed transport of intensity diffraction tomography with non-interferometric synthetic aperture (TIDT-NSA). Without resorting to interferometric detection, TIDT-NSA retrieves the 3D refractive index (RI) distribution of biological specimens from 3D intensity-only measurements at various illumination angles, allowing incoherent-diffraction-limited quantitative 3D phase-contrast imaging. The unique combination of z-scanning the sample with illumination angle diversity in TIDT-NSA provides strong defocus phase contrast and better optical sectioning capabilities suitable for high-resolution tomography of thick biological samples. Based on an off-the-shelf bright-field microscope with a programmable light-emitting-diode (LED) illumination source, TIDT-NSA achieves an imaging resolution of 206 nm laterally and 520 nm axially with a high-NA oil immersion objective. We validate the 3D RI tomographic imaging performance on various unlabeled fixed and live samples, including human breast cancer cell lines MCF-7, human hepatocyte carcinoma cell lines HepG2, mouse macrophage cell lines RAW 264.7, Caenorhabditis elegans (C. elegans), and live Henrietta Lacks (HeLa) cells. These results establish TIDT-NSA as a new non-interferometric approach to optical diffraction tomography and 3D label-free microscopy, permitting quantitative characterization of cell morphology and time-dependent subcellular changes for widespread biological and medical applications.


Heliox in children with croup: a strategy to hasten improvement.
Upper airway obstruction is responsive to the reduction in airflow turbulence provided by helium/oxygen (heliox) admixture. Our pediatric critical care transport team (PCCTT) has used heliox for children with upper airway obstruction from croup. We sought to describe our experience with heliox on transport and hypothesized that heliox-treated children with croup would show a more rapid clinical improvement. Children with croup transported by our PCCTT and admitted to the PICU were evaluated. We analyzed pretransport care, transport interventions, and outcomes. Croup scores (Modified Taussig) were assigned retrospectively according to respiratory therapy charting. Data were analyzed using appropriate statistical tests, including Pearson's chi-square test, Fisher's exact test, Mann-Whitney U rank comparison, and two-sample t-test. Thirty-five children met inclusion criteria. Demographics were similar between groups. The pretransport medical care was similar between groups. Children receiving heliox had a higher baseline croup score [mean (SD) = 5.7(2.3) vs no heliox 2.9 (2.0), P < 0.001]. The improvement in croup scores over the first 60 minutes of transport was more rapid in the heliox-treated children (P < 0.001). There was no difference in the number of children requiring additional nebulized racemic epinephrine during transport. The PICU length of stay (P = 0.59) and hospital length of stay (P = 0.64) were similar between groups. Heliox added to standard transport treatment for critically ill children with croup provides a more rapid improvement in croup scores. Heliox for croup during transport does not prolong intensive care unit stay. A prospective clinical trial is warranted to evaluate heliox in pediatric transport.


Optimization and efficient purification of recombinant Omp28 protein of Brucella melitensis using Triton X-100 and β-mercaptoethanol.
The high level expression of recombinant proteins in Escherichia coli often leads to the formation of inclusion bodies that contain most of the expressed protein held together by non-covalent forces. The inclusion bodies are usually solubilized using strong denaturing agents like urea and guanidium hydrochloride. In this study recombinant Omp28 (rOmp28) protein of Brucella melitensis was expressed in two different vector systems and further efficient purification of the protein was done by modification in buffers to improve the yield and purity. Different concentrations of Triton X-100 and β-mercaptoethanol were optimized for the solubilization of inclusion bodies. The lysis buffer with 8M urea alone was not sufficient to solubilize the inclusion bodies. It was found that the use of 1% Triton X-100 and 20mM β-mercaptoethanol in lysis and wash buffers used at different purification steps under denaturing conditions increased the yield of purified rOmp28 protein. The final yield of purified protein obtained with modified purification protocol under denaturing conditions was 151 and 90mg/l of the culture or 11.8 and 9.37mg/g of wet weight of cells in pQE30UA and pET28a(+) vector respectively. Thus modified purification protocol yielded more than threefold increase of protein in pQE30UA as compared with purification by conventional methods.


Smoking-related prospective memory deficits observed on naturalistic everyday memory task.
This study assessed self-reported and objective prospective memory (PM) processes in smokers and a never-smoked comparison group. If persistent smoking does impair PM, then one would expect smokers recall being lower on a study that requires them to remember everyday activities when compared with a never-smoked group. An existing-groups design was used to compare a group of smokers with a never-smoked group on the self-report Prospective Memory Questionnaire (PMQ) and the Prospective Remembering Video Procedure (PRVP) measuring objective PM. An example of the location-action combination from the PRVP is 'At Thornton's shop' (location), 'Buy a bag of sweets' (action). Participants who reported using an illegal substance (e.g. ecstasy, cannabis), who drank excessively or were 'binge drinkers', or who reported suffering from a clinical condition, such as depression, were excluded from the study. Age, weekly 'safe levels' alcohol use, and strategy use were also measured and controlled for in the study. Each person was tested individually in a quiet laboratory setting on a university campus. After controlling for variations in age, weekly alcohol use, and strategy use, smokers recalled significantly fewer location-action combinations on the PRVP when compared with a never-smoked group, with no between-group differences on self-reported PM as measured by the PMQ. The findings suggest objective PM deficits are associated with persistent smoking - a relatively unexplored area of research. This cannot be attributed to other drug use, mood, or strategy use. The findings also suggest smokers lack self-awareness of such PM deficits. This study extends the area by utilising a more naturalistic object measure of PM and incorporating strict controls into the study.


Virologic and clinical characteristics of HBV genotypes/subgenotypes in 487 Chinese pediatric patients with CHB.
The association of hepatitis B virus (HBV) genotypes/subgenotypes with clinical characteristics is increasingly recognized. However, the virologic and clinical features of HBV genotypes/subgenotypes in pediatric patients remain largely unknown. Four hundred and eighty-seven pediatric inpatients with CHB were investigated, including 217 nucleos(t)ide analog-experienced patients. HBV genotypes/subgenotypes and reverse transcriptase (RT) mutations were determined by direct sequencing. The stage of fibrosis and degree of inflammatory activity were evaluated by the Metavir score system. Among 487 enrolled pediatric patients, HBV genotype C2 and B2 were the most two prevalent (73.7% and 21.1%). Comparing with HBV/B2 infected patients, no significant difference was observed in the incidence rate and mutant patterns of lamivudine- or adefovir-resistant mutations in HBV/C2 infected patients (P > 0.05). Importantly, we found that the degree of hepatic inflammation degree, fibrosis stage and ALT level were significantly higher in HBV/C2-infected HBeAg positive patients than it was in HBV/B2-infected ones. The pediatric patients with HBV/C2 infection might be more susceptible to develop severe liver pathogenesis.


Usefulness of Postnatal Echocardiograms in Patients with Omphaloceles Who Previously Had a Normal Fetal Echocardiogram.
Neonates with omphaloceles routinely have a transthoracic echocardiogram (TTE) performed due to a high association with congenital heart defects (CHD). The utility of a TTE in these patients with a normal fetal echocardiogram is unknown. The primary objective of this study was to determine whether a critical CHD diagnosis was missed in patients with an omphalocele who had a normal fetal echocardiogram. The secondary objective of the study was to determine whether any CHD diagnosis was missed postnatally when a fetal echocardiogram was read as normal. A retrospective chart review was performed of patients with omphaloceles born between January 1, 2008, and June 30, 2020. Patients were included if they had a fetal echocardiogram that was read as normal and had a postnatal echocardiogram performed. Baseline demographics, postnatal data echocardiographic findings, and hospital course were collected. Critical CHD was defined as CHD requiring neonatal cardiac intervention. Fifty-six fetal echocardiograms on patients with omphaloceles were performed, of which 24 patients met the inclusion criteria. No patient was diagnosed with a critical CHD postnatally (negative predictive value [NPV] = 100%). Two patients were diagnosed with ventricular septal defects (VSD) postnatally (NPV = 91.7%). One of the VSDs required closure with a patch at 4 months of life, while the other, a small muscular VSD, closed spontaneously within the first year of life. Both patients had a murmur on exam during their initial hospital stay. The patient that required surgery also had an abnormal electrocardiogram and chest X-ray. There were no mortalities due to cardiac causes in these patients. Critical CHD was not missed on any patient with an omphalocele who had a normal fetal echocardiogram. All other patients with omphaloceles who had CHD diagnosed postnatally had an abnormal clinical finding on postnatal evaluation. The routine performance of a postnatal TTE in patients with an omphalocele who had a normal fetal echocardiogram may not be needed in those with a normal clinical workup. Further studies evaluating echocardiographic imaging recommendations are needed to maximize care and optimize resource allocation in this complex patient population.


Effect of zearalenone on aflatoxin B1-induced intestinal and ovarian toxicity in pregnant and lactating rats.
Aflatoxin B1 (AFB1) and zearalenone (ZEN) cause serious damage to mammals, but few studies have investigated the impacts of these toxins on pregnant and lactating mammals. This study investigated the effects of ZEN on AFB1-induced intestinal and ovarian toxicity in pregnant and lactating rats. Based on the results, AFB1 reduces the digestion, absorption, and antioxidant capacity in the intestine, increases intestinal mucosal permeability, destroys intestinal mechanical barriers, and increases pathogenic bacteria' relative abundances. Simultaneously, ZEN can exacerbate the intestinal injury caused by AFB1. The intestines of the offspring were also damaged, but the damage was less severe than that observed for the dams. While AFB1 activates various signalling pathways in the ovary and affects genes related to endoplasmic reticulum stress, apoptosis, and inflammation, ZEN may exacerbate or antagonize the AFB1 toxicity on gene expression in the ovary through key node genes and abnormally expressed genes. Our study found that mycotoxins can not only directly damage the ovaries and affect gene expression in the ovaries but can also impact ovarian health by disrupting intestinal microbes. Mycotoxins are an important environmental pathogenic factor for intestinal and ovarian disease in pregnancy and lactation mammals.


Ablation of matrix metalloproteinase-9 increases severity of viral myocarditis in mice.
Coxsackievirus B3 (CVB3) causes human myocarditis, which can result in cardiac damage, maladaptive remodeling, and heart failure. Matrix metalloproteinases (MMP)-8 and -9 have been identified in virus-infected myocardium, but their particular roles and underlying mechanisms of effect are unknown. For the first time, we examine the severity of CVB3-induced myocarditis in MMP-8-and MMP-9-deficient mice. CVB3-infected MMP-8 and MMP-9 knockout (KO) mice and corresponding wild-type (WT) mice were euthanized and harvested at 9 days after infection. Expression of MMP-2, -8, -12, and -13 and tissue inhibitors of MMPs was assessed by zymography or immunoblotting on harvested hearts, and in situ hybridization was performed to detect active infection. Infected MMP-9 KO mice had greater myocardial injury and foci of infection than WT mice despite similar pancreatic infection. Increased fibrosis (10.6+/-2.7% versus 7.1+/-2.6%, P=0.04), viral titer, as well as decreased cardiac output, were evident in MMP-9 KO compared with WT mice as assessed by picrosirius red staining, plaque assay, and echocardiography, respectively. Immune infiltration was also greatly increased in MMP-9 KO compared with WT mice (15.2+/-12.6% versus 2.0+/-3.0%, P<0.002). Myocardial interferon-beta1, interferon-gamma, interleukin-6, interleukin-10, and macrophage inflammatory protein-1alpha expression was elevated in MMP-9 KO mice as measured by quantitative real-time polymerase chain reaction and ELISA. In contrast, MMP-8 KO mice had the same degree of cardiac injury, fibrosis, and viral infection as their WT counterparts. During acute CVB3 infection, MMP-9 appears necessary to halt virus propagation in the heart, promote proper immune infiltration and remodeling, and preserve cardiac output.


A SOCS1/3 Antagonist Peptide Protects Mice Against Lethal Infection with Influenza A Virus.
We have developed an antagonist to suppressor of cytokine signaling 1 (SOCS1), pJAK2(1001-1013), which corresponds to the activation loop of the Janus kinase JAK2, which is the binding site for the kinase inhibitory region (KIR) of SOCS1. Internalized pJAK2(1001-1013) inhibits SOCS1 and SOCS3. SOCS1 has been shown to be an influenza virus-induced virulence factor that enhances infection of cells. The antagonist was protective in cell culture and in influenza virus PR8 lethally infected C57BL/6 mice. The SOCS antagonist also prevented adverse morbidity as assessed by parameters, such as weight loss and drop in body temperature, and showed potent induction of both the cellular and humoral immune responses to the influenza virus candidate universal antigen matrix protein 2 (M2e). The SOCS antagonist, thus, protected mice against lethal influenza virus infection and possessed potent adjuvancy against the M2e candidate influenza virus universal vaccine antigen.


On the canonical energy of weak gravitational fields with a cosmological constant Λ ∈ R.
We analyse the canonical energy of vacuum linearised gravitational fields on light cones on a de Sitter, Minkowski, and Anti de Sitter backgrounds in Bondi gauge. We derive the associated asymptotic symmetries. When Λ > 0 the energy diverges, but a renormalised formula with well defined flux is obtained. We show that the renormalised energy in the asymptotically off-diagonal gauge coincides with the quadratisation of the generalisation of the Trautman-Bondi mass proposed in Chruściel and Ifsits (Phys Rev D 93:124075, arXiv:1603.07018 [gr-qc], 2016).


Amyloid precursor protein and amyloid beta-peptide bind to ATP synthase and regulate its activity at the surface of neural cells.
Amyloid precursor protein (APP) and amyloid beta-peptide (Abeta) have been implicated in a variety of physiological and pathological processes underlying nervous system functions. APP shares many features with adhesion molecules in that it is involved in neurite outgrowth, neuronal survival and synaptic plasticity. It is, thus, of interest to identify binding partners of APP that influence its functions. Using biochemical cross-linking techniques we have identified ATP synthase subunit alpha as a binding partner of the extracellular domain of APP and Abeta. APP and ATP synthase colocalize at the cell surface of cultured hippocampal neurons and astrocytes. ATP synthase subunit alpha reaches the cell surface via the secretory pathway and is N-glycosylated during this process. Transfection of APP-deficient neuroblastoma cells with APP results in increased surface localization of ATP synthase subunit alpha. The extracellular domain of APP and Abeta partially inhibit the extracellular generation of ATP by the ATP synthase complex. Interestingly, the binding sequence of APP and Abeta is similar in structure to the ATP synthase-binding sequence of the inhibitor of F1 (IF(1)), a naturally occurring inhibitor of the ATP synthase complex in mitochondria. In hippocampal slices, Abeta and IF(1) similarly impair both short- and long-term potentiation via a mechanism that could be suppressed by blockade of GABAergic transmission. These observations indicate that APP and Abeta regulate extracellular ATP levels in the brain, thus suggesting a novel mechanism in Abeta-mediated Alzheimer's disease pathology.


A fatal case of acute HHV-6 myocarditis following allogeneic haemopoietic stem cell transplantation.
Human herpesvirus 6 (HHV-6) is an ubiquitous virus that can reactivate in immunocompromised hosts, resulting in diverse clinical sequelae. We describe a case of fatal acute HHV-6 myocarditis in a patient who underwent allogeneic haemopoietic stem cell transplantation (HSCT). To our knowledge, this is the first reported case of biopsy proven HHV-6 myocarditis post-HSCT.


Prenatal closure of the foramen ovale in complete transposition of great vessels. Observations in a case with intact ventricular septum and normally developed left heart chambers.
A rare case is reported of premature closure of the foramen ovale in d-transposition of the great vessels with intact ventricular septum and normal development of the left side of the heart. Asphyxia neoatorum and cardiac failure were the manifestations in the early neonatal period. Balloon atrial septostomy during cardiac catheterization would be unsuccessful in infants with this condition, and closed atrial septectomy (Blalock-Hanlon procedure) would be necessary for survival.


Effect of ageing on surface IgG of human peripheral lymphocytes.
An increase in the percentage of IgG bearing peripheral blood lymphocytes is observed in aged subjects as compared with young ones. Such a finding is probably due to the presence, in 'aged' sera, of a higher concentration of immune complexes, bound to lymphocytes through their Fc or Complement receptors.


Pharmacokinetics and metabolism of bepridil.
Bepridil hydrochloride differs from the other calcium antagonists in structure as well as in several clinical pharmacokinetic characteristics. The drug is completely absorbed from the gastrointestinal tract, but first-pass extraction reduces oral bioavailability to approximately 60%. After single-dose administration, the elimination half-life of bepridil averages 33 +/- 15 hours. However, upon multiple dosing, a half-life of 42 +/- 12 hours is found. As with verapamil and diltiazem, bepridil clearance is decreased after multiple dosing. Bepridil is completely metabolized, presumably by hepatic oxidative processes. A total of 17 metabolites have been identified, but the contribution of any of these metabolites to observed clinical response is currently unclear. The free fraction of bepridil in plasma is low, averaging only 0.23%. Despite this high protein binding, in vitro studies indicate that the potential for drug-to-drug interactions based on displacement of bepridil from its binding sites is low. Bepridil follows a linear dose/plasma concentration relation after single and multiple doses of the drug in both healthy volunteers and patients with angina. However, mean steady-state plasma bepridil concentrations are higher in patients, indicating a greater average decreased clearance. Food does not interfere with bepridil absorption. At this time, no significant pharmacokinetic interactions between bepridil and digoxin have been detected.


The effect of increased inferior vena cava pressure on hepatic circulation in the dog.
Inferior v. cava pressure (IVCP) was raised in the dog by a baloon catheter introduced via the jugular vein. Hepatic artery flow (HAF) and portal venous flow (PVF) were measured with the electromagnetic flow meter. The increase of IVCP reduced uniformly HAF and PVF, the relative contribution of the two vessels to the HBF did not change. The relationship between IVCP increase and HAF reduction and between the decrease of AP-IVCP and the reduction of HAF was linear. There was no sign of autoregulation or vasomotor regulation of HAF. The changes in IVCP are only partially transmitted to the portal venous pressure (PVP). There was no change in intrahepatic portal resistance with increased IVCP. The increase in PVP did not lead to resistance changes in the splanchnic circulation. As a sign of the autoregulation of splanchnic circulation of the AP-PVP versus PVF curve was concave to the pressure axis. It is concluded that as a result of opposing autoregulatory and vasomotor influences in the intact animal the hepatic circulation appears to react passively to the increased outflow pressure.


Necroptotic astrocytes contribute to maintaining stemness of disseminated medulloblastoma through CCL2 secretion.
Medulloblastoma (MB) with metastases at diagnosis and recurrence correlates with poor prognosis. Unfortunately, the molecular mechanism underlying metastases growth has received less attention than primary therapy-naïve MB. Though astrocytes have been frequently detected in brain tumors, their roles in regulating the stemness properties of MB stem-like cells (MBSCs) in disseminated lesions remain elusive. Effects of tumor-associated astrocyte (TAA)-secreted chemokine C-C ligand 2 (CCL2) on MBSC self-renewal was determined by immunostaining analysis. Necroptosis of TAA was examined by measuring necrosome activity. Alterations in Notch signaling were examined after inhibition of CCL2. Progression of MBSC-derived tumors was evaluated after pharmaceutical blockage of necroptosis. TAA, as the essential components of disseminated tumor, produced high levels of CCL2 to shape the inflammation microenvironment, which stimulated the enrichment of MBSCs in disseminated MB. In particular, CCL2 played a pivotal role in maintaining stem-like properties via Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3)-mediated activation of Notch signaling. Loss of CCL2/C-C chemokine receptor 2 (CCR2) function repressed the JAK2/STAT3-Notch pathway and impaired MBSC proliferation, leading to a dramatic reduction of stemness, tumorigenicity, and metastasizing capability. Furthermore, necroptosis-induced CCL2 release depended on activation of receptor-interacting protein 1 (RIP1)/RIP3/mixed lineage kinase domain-like pseudokinase (MLKL) in TAA, which promoted the oncogenic phenotype. Blockade of necroptosis resulted in CCL2 deprivation and compromised MBSC self-proliferation, indicating MBSCs outsourced CCL2 from necroptotic TAA. Finally, CCL2 was upregulated in high-risk stages of MB, further supporting its value as a prognostic indicator. These findings highlighted the critical role of CCL2/CCR2 in Notch signaling activation in MBSCs and revealed a necroptosis-associated glial cytokine microenvironment driving stemness maintenance in disseminations.Key Points1. TAA-derived CCL2 promoted stemness in disseminated MBSCs through Notch signaling activation via the JAK2/STAT3 pathway.2. TAA released CCL2 in a RIP1/RIP3/MLKL-dependent manner leading to necroptosis.


[Bladder tumor occluding diverticular orifice: a case with difficult diagnosis due to hemorrhage into diverticulum].
We experienced a case of bladder diverticular tumor with intradiverticular hemorrhage in which preoperative diagnosis was difficult. It simultaneously occurred with a bladder tumor. The patient, a 70-year-old man complained of macrohematuria and pollakisuria. The cystoscopic examination revealed a bladder tumor. The radiographic examination suggested a bladder tumor with extravesical hemorrhage. We diagnosed an invasive bladder tumor and performed radical cystectomy and ileal conduit diversion. Postoperative diagnosis was bladder tumor and diverticular tumor.


Prognostic value of HMGB3 expression in patients with non-small cell lung cancer.
HMGB3 overexpression has been reported in a variety of human cancers. However, the role of HMGB3 in human non-small cell lung cancer (NSCLC) remains unclear. In this study, the HMGB3 expression was examined at mRNA and protein levels by quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR), Western blotting, and immunohistochemistry in NSCLC tissues and adjacent non-cancerous tissues. Statistical analyses were applied to test the associations between HMGB3 expression, clinicopathologic factors, and prognosis. Western blotting and qRT-PCR showed that the expression levels of HMGB3 mRNA and protein were both significantly higher in NSCLC tissues than those in non-cancerous tissues. Immunohistochemistry analysis showed that HMGB3 expression was significantly correlated with tumor grade, tumor size, clinical stage, and lymph node metastases. The results of Kaplan-Meier analysis indicated that a high expression level of HMGB3 resulted in a significantly poor prognosis of NSCLC patients. Importantly, multivariate analysis showed that high HMGB3 expression was an independent prognostic factor for NSCLC patients. In sum, our data suggest that HMGB3 plays an important role in NSCLC progression, and that overexpression of HMGB3 in tumor tissues could be used as a potential prognostic marker for patients with NSCLC.


Lethal neuroleptic malignant syndrome due to amisulpride.
A 42-year old-man was found lying in his bed having seizures. Later he became unconscious and hypotonic developing mydriasis as well as rigidity. The body core temperature (rectal temperature) was above 42 °C. Blood pH was decreased during treatment, and his general condition deteriorated. The patient developed gasping respiration, ventricular fibrillation, and died. During autopsy and histological investigation cerebral and pulmonary edema were noted together with general congestion of the internal organs. Further observations included contraction bands of myocytes, a contracted spleen, fibrosis of the liver, and gall stones. Toxicological analyses of peripheral blood revealed the following results: amisulpride 4.65 mg/l, biperiden 0.12 mg/l, imipramine 0.33 mg/l, and desipramine 0.68 mg/l. An amisulpride-induced neuroleptic malignant syndrome was therefore diagnosed as the patho-physiological mechanism leading to death.


Budesonide in previously untreated autoimmune hepatitis.
Autoimmune hepatitis (AIH) is a chronic liver disease that is effectively treated with immunosuppressive therapy. Predniso(lo)ne, often in combination with azathioprine, is the basic therapeutic option to induce remission. However, this regimen can cause numerous side effects. The aim of the present study was to evaluate budesonide as a treatment option in the induction of remission in patients with previously untreated AIH. Between October 1998 and August 1999, 12 patients were treated with 3 mg budesonide thrice daily for 3 months in this open one-arm multicenter phase IIa study. Primary end point was induction of remission indicated by a drop of aspartate aminotransferase and alanine aminotransferase levels below two times the upper limit of normal. Seven of the 12 patients (58%) reached complete remission, three patients (25%) had a partial response. Thus, 10/12 individuals (83.3%) responded to therapy. Therapy was tolerated well in 10/12 cases (83.3%). Budesonide monotherapy was effective in the induction of remission and well tolerated in treatment naïve patients with AIH. It should be further evaluated in prospective controlled trials and should be compared to predniso(lo)ne both in monotherapy and in combination with azathioprine.


Association of Microtubule Dynamics with Chronic Epilepsy.
Approximately 30 % of epilepsy cases are refractory to current pharmacological treatments through unknown mechanisms. Much work has been done on the role of synaptic components in the pathogenesis of epilepsy, but relatively little attention has been given to the potential role of the microtubules. We investigated the level of microtubule dynamic in 30 human epileptic tissues and two different chronic epilepsy rat models. The administration of microtubule-modulating agent attenuated the progression of chronic epilepsy. By contrast, microtubule-depolymerizing agent aggravated the progression of chronic epilepsy. The electrophysiological index by whole-cell clamp was used to investigate the neuronal excitation and inhibitory synaptic transmission in brain slices after administration of microtubule-modulating agent and microtubule-depolymerizing agent. Interestingly, we found that microtubule-modulating agent significantly increased the frequency of action potential firing in interneurons, and significantly promoted the amplitudes and frequencies of miniature inhibitory postsynaptic currents. Microtubule-depolymerizing agent had an opposite effect. These findings suggest that modulating hyperdynamic microtubules may take an anti-epileptic effect via postsynaptic mechanisms in interneurons. It could represent a potential pharmacologic target in epilepsy treatment.


Molecular basis for cysteine oxidation by plant cysteine oxidases from Arabidopsis thaliana.
Plant Cysteine Oxidases (PCOs) play important roles in controlling the stability of Group VII ethylene response factors (ERF-VIIs) via Arg/N-degron pathway through catalyzing the oxidation of their N-Cys for subsequent Arginyl-tRNA--protein transferase 1 (ATE1) mediated arginine installation. Here we presented the crystal structures of PCO2, PCO4, and PCO5 from Arabidopsis thaliana (AtPCOs) and examined their in vitro activity by Mass spectrometry (MS). On the basis of Tris-bound AtPCO2, we modelled the structure of Cys-bound AtPCO2 and identified key AtPCO2 residues involved in N-Cys recognition and oxidation. Alanine substitution of potential N-Cys interaction residues impaired the activity of AtPCO5 remarkably. The structural research, complemented by mutagenesis and MS experiments, not only uncovers the substrate recognition and catalytic mode by AtPCOs, but also sheds light on the future design of potent inhibitors for plant cysteine oxidases.


Cyclops syndrome occurring after chronic partial rupture of the anterior cruciate ligament without surgical reconstruction.
Cyclops syndrome is one of the specific causes of loss of extension of the knee following anterior cruciate ligament (ACL) reconstruction. This syndrome is manifest by progressive loss of knee extension associated with pain and audible clunk at terminal extension caused by a pedunculated nodule of fibrovascular proliferative tissue usually arising from a graft. Recent published reports, however, have described Cyclops syndrome also developing after a partial ACL rupture without surgical reconstruction. In most cases, Cyclops syndrome generally occurred within 2 months after reconstruction surgery or rupture. Here we report on the case of a patient with symptoms and arthroscopic and histological findings compatible with Cyclops syndrome that developed after a chronic partial ACL rupture that occurred 23 years ago. To our knowledge, this is the first case report of Cyclops syndrome occurring after chronic partial ACL rupture.


Differential gene expression induced by growth hormone treatment in the uremic rat growth plate.
Treatment with growth hormone (GH) improves growth retardation of chronic renal failure. cDNA microarrays were used to investigate GH-induced modifications in gene expression in the tibial growth plate of young rats. RNA was extracted from the tibial growth plate from two groups, untreated and treated with GH, of young rats made uremic by subtotal nephrectomy (n=10). To validate changes shown by the Agilent oligo microarrays, some modulated genes known to play a physiological role in growth plate metabolism were analyzed by real-time quantitative polymerase chain reaction (qPCR). The microarrays showed that GH modified the expression of 224 genes, 195 being upregulated and 29 downregulated. qPCR results confirmed the sense of expression change found in the arrays for insulin-like growth factor I, insulin-like growth factor II, collagen V alpha 1, bone morphogenetic protein 3 and proteoglycan type II. This study shows for the first time the profile of growth plate gene expression modifications caused by GH treatment in experimental uremia and provides a basis to further investigate selected individual genes with potential implication in the stimulating effect on the growth of GH treatment in chronic renal failure.


A multi-granularity convolutional neural network model with temporal information and attention mechanism for efficient diabetes medical cost prediction.
As the cost of diabetes treatment continues to grow, it is critical to accurately predict the medical costs of diabetes. Most medical cost studies based on convolutional neural networks (CNNs) ignore the importance of multi-granularity information of medical concepts and time interval characteristics of patients' multiple visit sequences, which reflect the frequency of patient visits and the severity of the disease. Therefore, this paper proposes a new end-to-end deep neural network structure, MST-CNN, for medical cost prediction. The MST-CNN model improves the representation quality of medical concepts by constructing a multi-granularity embedding model of medical concepts and incorporates a time interval vector to accurately measure the frequency of patient visits and form an accurate representation of medical events. Moreover, the MST-CNN model integrates a channel attention mechanism to adaptively adjust the channel weights to focus on significant medical features. The MST-CNN model systematically addresses the problem of deep learning models for temporal data representation. A case study and three comparative experiments are conducted using data collected from Pingjiang County. Through experiments, the methods used in the proposed model are analyzed, and the super contribution of the model performance is demonstrated.


One-Pot Synthesis of Nitrogen-Doped Graphene Quantum Dots and Their Applications in Bioimaging and Detecting Copper Ions in Living Cells.
Two natural carbon sources, glutamic acid and tyrosine, were used to fabricate strong green emission nitrogen-doped graphene quantum dots (N-GQDs) with the one-pot pyrolysis method. The morphology of the prepared GQDs has been characterized by high-resolution transmission electron microscopy, showing a well-displayed crystalline structure with a lattice spacing of 0.262 nm. X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy were used to analyze the surface functional groups and elemental composition, suggesting that the N-GQDs have active carboxylic and amino functional groups. Meanwhile, photoluminescence and ultraviolet-visible (UV-vis) spectroscopy were used to evaluate the optical properties of GQDs; the prepared N-GQDs show an excitation-dependent fluorescence behavior with a maximum excitation/emission wavelength at 460/522 nm, respectively. N-GQDs showed good photostability and the fluorescence intensity quenched about 10% after irradiating 2800 s in the experiment of time kinetic analysis. The MTT assay was utilized to assess the viability of N-GQDs; good biocompatibility with a relatively high quantum yield of 12% demonstrated the potential for serving as bioimaging agents. Besides, the selectivity study on metal ions indicates that the N-GQDs could be used in Cu2+ detection. The linear range is from 0.1 to 10 μM with a limit of detection of 0.06 μM. Overall, these proposed N-GQDs with one-pot synthesis showed their promising potential in cell imaging and Cu2+ monitoring applications involved in the biological environment.


Modified forward-backward splitting midpoint method with superposition perturbations for the sum of two kinds of infinite accretive mappings and its applications.
In a real uniformly convex and p-uniformly smooth Banach space, a modified forward-backward splitting iterative algorithm is presented, where the computational errors and the superposition of perturbed operators are considered. The iterative sequence is proved to be convergent strongly to zero point of the sum of infinite m-accretive mappings and infinite [Formula: see text]-inversely strongly accretive mappings, which is also the unique solution of one kind variational inequalities. Some new proof techniques can be found, especially, a new inequality is employed compared to some of the recent work. Moreover, the applications of the newly obtained iterative algorithm to integro-differential systems and convex minimization problems are exemplified.


Paxlovid administration in elderly patient with COVID-19 caused by Omicron BA.2.0: A case report.
Paxlovid has shown the potential decreasing the hospitalization rate of mild or moderate coronavirus disease 2019 (COVID-19) and death in few of clinical trials, and is expected to the most promising medicine targeting Severe Acute Respiratory Syndrome Coronavirus 2 (SRAS-COV-2). However, there are no enough evidences to show it effectiveness for all patients with SARS-COV-2, especially among elderly patients and newest Omicron variant. A 79 year's old female patient was admitted to hospital because of the moderate COVID-19 caused by the Omicron variant BA2.0. He presented the initial syndromes including Xerostomia, cough and fever. Chest computed tomography (CT) scanning at admission showed the exudation lesions on lung. The laboratory examination revealed that there are increased C-reactive protein (CRP), Ferritin and erythrocytesedimentationrate (ESR) and decreased white blood cells. The oral Paxlovid (Nirmatrelvir/Ritonavir) was administrated on second day after admission. The syndromes of Xerostomia, cough and fever was improved on third day after use of Paxlovid. The levels of CRP, ESR and counts of white blood cells returned the normal after three days of admission. The chest CT scanned on the third and sixth day after Paxlovid used showed the absorption of lesions. The examination of SARS-COVS viral nucleic acid turned negative at fifth day of admission. As a result, we would consider that Paxlovid is a suitable oral drug for elderly patients with SARS-COV2 even Omicron variant, it's benefit to improve patient's symptom and signs and can prevents COVID-19 with the high-risk factors from severe disease, although it didn't shorten the time for viral nucleic acid to turn negative.


Erratum: Random Field Driven Spatial Complexity at the Mott Transition in VO_{2} [Phys. Rev. Lett. 116, 036401 (2016)].
This corrects the article DOI: 10.1103/PhysRevLett.116.036401.


Evaluation of selected biomarkers for the detection of chemical sensitization in human skin: a comparative study applying THP-1, MUTZ-3 and primary dendritic cells in culture.
Dendritic cells (DCs) exhibit the unique capacity to induce T cell differentiation and proliferation, two processes that are crucially involved in allergic reactions. By combining the exclusive potential of DCs as the only professional antigen-presenting cells of the human body with the well known handling advantages of cell lines, cell-based alternative methods aimed at detecting chemical sensitization in vitro commonly apply DC-like cells derived from myeloid cell lines. Here, we present the new biomarkers programmed death-ligand 1 (PD-L1), DC immunoreceptor (DCIR), IL-16, and neutrophil-activating protein-2 (NAP-2), all of which have been detectable in primary human DCs upon exposure to chemical contact allergens. To evaluate the applicability of DC-like cells in the prediction of a chemical's sensitization potential, the expression of cell surface PD-L1 and DCIR was analyzed. In contrast to primary DCs, only minor subpopulations of MUTZ-3 and THP-1 cells presented PD-L1 or DCIR at their surface. After exposure to increasing concentrations of nickel and cinnamic aldehyde, the expression level of PD-L1 and DCIR revealed much stronger affected on monocyte-derived DCs (MoDCs) or Langerhans cells (MoLCs) when compared to THP-1 and MUTZ-3 cells. Applying protein profiler arrays we further identified the soluble factors NAP-2, IL-16, IL-8 and MIP-1α as sensitive biomarkers showing the capacity to discriminate sensitizing from non-sensitizing chemicals or irritants. An allergen-specific release of IL-8 and MIP-1α could be detected in the supernatants of MoDCs and MoLCs and also in MUTZ-3 and THP-1 cells, though at much lower levels. On the protein and transcriptional level, NAP-2 and IL-16 indicated sensitizers most sensitively and specifically in MoDCs. Altogether, we have proven the reciprocal regulated surface molecules PD-L1 and DCIR and the soluble factors MIP-1α, NAP-2 and IL-16 as reliable biomarkers for chemical sensitization. We further show that primary DCs are significantly different in their phenotype and function compared to DC-like cell lines. Since they demonstrated higher absolute values and a broader range in biomarker expression, we propose that MoDCs represent an optimal and robust sensor test system well suited to identify and classify chemicals with an allergic potential.