Dabrafenib in an elderly patient with metastatic melanoma and BRAF V600R mutation: a case report.
Approximately 50 % of malignant melanomas harbor activating point mutations in the BRAF gene. Typically, these mutations result in the substitution of the amino acid valine at codon 600 of the gene, and 90-95 % of mutations are either BRAF (V600E) or BRAF (V600K). Specific BRAF inhibitors such as dabrafenib and vemurafenib are the mainstays of treatment in patients with metastatic BRAF-mutant malignant melanomas. The third most common BRAF mutation is V600R, which also leads to increased BRAF signaling. Although evidence exists about the activity of dabrafenib and vemurafenib in patients with the BRAF (V600R) mutation, these patients have been systematically excluded from recent trials with targeted therapies. Here, we report the positive results in terms of survival and quality of life obtained with dabrafenib in an 80-year-old Caucasian male patient with a Charlson Comorbidity Index of 8 diagnosed with metastatic malignant melanoma harboring the BRAF (V600R) mutation. Our patient was treated with dabrafenib for 7 months with minimal toxicity. We also report exploratory analyses of circulating tumor DNA during targeted treatment. Interestingly, the mutation was not detected after starting treatment and became detectable before radiological disease progression. Our report suggests that (1) a relevant benefit can be obtained with a BRAF inhibitor in real-world patients with a malignant melanoma harboring a BRAF (V600R) mutation, and that (2) circulating tumor DNA detection might be of help in assessing tumor burden in everyday clinical practice. The results reported here should encourage the inclusion of patients with BRAF (V600R)-mutated malignant melanomas in future prospective clinical trials with BRAF inhibitors.


Cystic ameloblastoma. A clinicopathologic analysis.
Thirty-three cases of ameloblastoma arising from odontogenic cyst linings are presented. Lesions of this nature have been referred to as "unicystic ameloblastoma." Our data support the findings of others in that this lesion differs from the solid invasive varieties of ameloblastoma, as the former occurs in a younger age group and exhibits a low rate of recurrence following enucleation or curettage. On the basis of histopathologic features we provide evidence that these lesions represent mural and luminal ameloblastomatous change in a pre-existing cyst rather than cystic degeneration of a solid tumor.


Mg2+ modulates membrane sphingolipid and lipid second messenger levels in vascular smooth muscle cells.
In vitro studies with smooth muscle cells from rat aorta and dog cerebral blood vessels indicate that variation in free Mg2+, within the pathophysiological range of Mg2+ concentrations, found in human serum, causes sustained changes in membrane phospholipids and lipid second messengers. Incorporation of [3H]palmitic acid into phosphatidylcholine (PC) and sphingomyelin (SM) was altered within 15-30 min after modifying the extracellular Mg2+ ion level ([Mg2+]o). Decreased Mg2+ produced a fall in both [3H]SM and [3H]PC over the first 2 h. After an 18-h incubation, the [3H]PC/[3H]SM ratio changed from about 20:1 to about 50:1. Increased [Mg2+]o resulted in a 2- to 3-fold increase in [3H]SM compared to only a small increase in [3H]PC over the same period. There was a reciprocal relationship between [3H]ceramide and [3H]1,2-DAG levels with highest [3H]ceramide and lowest [3H]-1,2-DAG levels seen at lowest [Mg2+]o. The results indicate that a fall in extracellular ionized Mg2+ concentration produces a rapid and sustained decrease in membrane sphingomyelin and a moderate rise in intracellular ceramide. A major effect of lowering [Mg2+]o appears to be a down-regulation of SM synthase. The increased membrane SM content and a concomitant decrease in cell ceramide, in the presence of elevated [Mg2+]o, may be relevant to the apparent protective role of adequate Mg intake on vascular function in humans.


NIR spectroscopic methods for monitoring blend potency in a feed frame - calibration transfer between offline and inline using a continuum regression filter.
A material sparing method for near-infrared (NIR) calibration was developed using an offline apparatus coupled with a calibration transfer method to enable a partial least squares (PLS) model to monitor the concentration of active pharmaceutical ingredients (API) in the feed frame of a rotary tablet press. The offline apparatus was designed to simulate the powder flow dynamic and NIRS measurement environment of a tablet-press feed frame. A comprehensive experimental design, including calibration and testing, was employed to determine blend inhomogeneity. NIR spectra were collected at both the feed frame (inline) conditions and the simulator (offline) conditions. The simulator conditions were designed to mimic the density and powder flow in the feed frame during the actual tableting process. The offline data were pretreated by an orthogonalization-based calibration transfer algorithm, a continuum regression filter (CR filter), before being subjected to PLS modeling. This study demonstrated: (1) calibration for inline application can be generated using an offline apparatus, and (2) the CR filter, as an innovative calibration transfer method, can generalize the offline method for multiple feed frame conditions.


Prediction of facial nerve displacement in extra large vestibular schwannoma.
The primary objective in the surgery of extra large vestibular schwannoma is the total removal of the tumour mass while preserving the facial nerve. Preservation of the facial nerve in extra large tumours is reported as being notoriously difficult in the majority of cases. This study was undertaken to evaluate the accuracy in predicting displacement of the facial nerve by preoperative radiological imaging studies in 19 cases of extra large vestibular schwannoma. The direction of displacement of the facial nerve was predicted with preoperative axial and coronal MRI scans and verified intraoperatively. We achieved total removal of tumours in 84.2%, facial nerve displacement was predicted in 80% and we accomplished anatomical preservation in 80%. Prediction of displacement was difficult in tumours with little or no intracanalicular components or with severe bony destruction of the internal acoustic meatus. The preoperative prediction of facial nerve displacement in extra large tumours allows safe internal decompression of the tumour and careful dissection near the predicted area of the facial nerve during the operation. Consequently, a high rate of anatomical preservation of the facial nerve can be achieved.


Ca2+ transport in mitochondria of the ciliate protozoan Tetrahymena pyriformis.
Mitochondria isolated from the late-exponential non-shaken culture of the ciliate protozoan Tetrahymena pyriformis GL was investigated. The presence of energy-dependent Ca2+ transport system was shown. In the main the properties of this system have been essentially the same as in mitochondria of vertebrate organisms. The isolated mitochondria contained 23 +/- 5 ng-ion Ca2+ per mg of protein. The intramitochondrial free concentration of Ca2+ was measured in the presence of uncoupler FCCP with the use of fluorescent Ca2+ chelator chlortetracycline and null point titration method. In the absence of phosphate, free [Ca2+] varied from 1 to 2.5 mM depending on the internal Ca2+ content. In the presence of 2 mM phosphate, free [Ca2+]in has not exceeded 0.1-0.3 mM. It was shown that ruthenium red and Mg2+ in different manner have an inhibitory effect on Ca2+ transport. Besides this, Mg2+ also has a stabilizing effect on mitochondria, possibly, by preventing passive ions leaks across the membrane.


Isolation of an Acinetobacter sp. ST-550 which produces a high level of indigo in a water-organic solvent two-phase system containing high levels of indole.
Acinetobacter sp. strain ST-550 was isolated from fumus soil as an efficient indigo producer in the presence of organic solvent. The minimum inhibitory concentration of indole was 0.4 mg/ml for ST-550. ST-550 produced only a small amount of indigo (less than 0.01 microg/ml) when grown in the presence of indole at concentrations of 0.05 to 0.3 mg/ml without any organic solvent. However. ST-550 produced indigo effectively when grown in the presence of a large volume of diphenylmethane and a high level of indole: optimized conditions were 3 ml of a medium containing 0.3 ml diphenylmethane and 2.7 mg indole. Under these conditions, ST-550 produced 0.88 mg indigo (292 microg/mI medium).


Inflammation-induced repression of chromatin bound by the transcription factor Foxp3 in regulatory T cells.
The transcription factor Foxp3 is indispensable for the ability of regulatory T cells (Treg cells) to suppress fatal inflammation. Here we characterized the role of Foxp3 in chromatin remodeling and the regulation of gene expression in actively suppressive Treg cells in an inflammatory setting. Although genome-wide occupancy of regulatory elements in DNA by Foxp3 was similar in resting Treg cells and those activated in vivo, Foxp3-bound enhancer elements in the DNA were poised for repression only in activated Treg cells. Following activation, Foxp3-bound sites showed diminished accessibility of chromatin and selective deposition of histone H3 trimethylated at Lys27 (H3K27me3), which was associated with recruitment of the histone methyltransferase Ezh2 and downregulation of the expression of nearby genes. Thus, Foxp3 poises its targets for repression by facilitating the formation of repressive chromatin in Treg cells upon their activation in response to inflammatory cues.


Role of depression and inflammation in incident coronary heart disease events.
Inflammatory biomarkers and depression have been proposed as novel coronary heart disease (CHD) risk markers. However, prospective studies have rarely assessed these 2 candidate CHD risk markers simultaneously in predicting incident CHD events. Therefore, although depression and elevated inflammatory biomarkers frequently covary, it is unclear how these risk markers relate to each other and to CHD event onset. The elucidation of these causal pathways has important clinical implications for patients who are depressed and/or have elevated inflammatory biomarkers. In this review, the publications examining the relations among depression, inflammation, and CHD events are discussed.


An evaluation of invasive fungal sinusitis outcomes with subsite analysis and use of frozen section analysis.
Invasive fungal sinusitis (IFS) is an aggressive mycosis of the nasal cavity with frequent extension to adjacent structures. Occurring more commonly in immunocompromised individuals, prognosis is typically poor despite aggressive treatment. This study aims to examine postoperative outcomes and survival of a cohort of fungal sinusitis patients at an academic center, as well as identify causes of death in IFS patients. This study was a retrospective chart review of patient charts and departmental records, yielding patient demographics, medical and surgical treatments, pathology records, and outcomes data. Twenty-seven patients were identified from departmental records between 1998 and 2014. Twenty-one patients presented with Mucor infections, whereas the remaining 6 patients had Aspergillus. All patients were immunocompromised: diabetes (n = 14) and hematologic malignancy (n = 13). Three patients had multiple causes of immunosuppression. Most commonly involved subsites were the maxillary, ethmoid, and sphenoid sinuses. Nasal septum involvement was independently associated with mortality (p < 0.01). Overall mortality was 57.7% within 1 year, although 66.7% of fatalities occurred within 1 month of diagnosis. Overall survival for IFS remains poor. Widespread disease and nasal septum involvement were associated with a negative clinical course. Early identification and aggressive surgical and antifungal therapy is warranted. Even despite intense therapy, comorbid conditions and drug toxicity increase mortality and complicate the clinical course.


Meeting report - NSF-sponsored workshop 'Progress and Prospects of Single-Molecule Force Spectroscopy in Biological and Chemical Sciences'.
The goals of the workshop organized by Piotr Marszalek and Andres Oberhauser that took place between 29 August and 1 September 2019 at Duke University were to bring together leading experts and junior researchers to review past accomplishments, recent advances and limitations in the single-molecule force spectroscopy field, which examines nanomechanical forces in diverse biological processes and pathologies. Talks were organized into four sessions, and two in-depth roundtable discussion sessions were held.


Characterization of hemodynamic events following intravascular infusion of recombinant adenovirus reveals possible solutions for mitigating cardiovascular responses.
Intravascular administration of recombinant adenovirus (rAd) in cancer patients has been well tolerated. However, dose-limiting hemodynamic responses associated with suppression of cardiac output have been observed at doses of 7.5 x 10(13) particles. While analysis of hemodynamic responses induced by small-molecule pharmaceuticals is well established, little is known about the cardiovascular effects of rAd. Telemetric cardiovascular (CV) monitoring in mice was utilized to measure hemodynamic events following intravascular rAd administration. Electrocardiogram analysis revealed a block in the SA node 3-4 min postinfusion, resulting in secondary pacemaking initiated at the AV node. This was associated with acute bradycardia, reduced blood pressure, and hypothermia followed by gradual recovery. Adenovirus-primed murine sera with high neutralizing antibody (nAb) titers could inhibit CV responses, whereas human sera with equivalent nAb titers induced by natural infection were, surprisingly, not inhibitory. Interestingly, repeat dosing within 2-4 h of the primary injection resulted in desensitization, resembling tachyphylaxis, for subsequent CV responses. Last, depletion of Kupffer cells prior to rAd infusion precluded induction of CV responses. These inhibitory effects suggest that rAd interactions with certain cells of the reticular endothelial system are associated with induction of CV responses. Significantly, these studies may provide insight into management of acute adverse effects following rAd systemic delivery, enabling a broadening of therapeutic index.


Overview of artificial intelligence-based applications in radiotherapy: Recommendations for implementation and quality assurance.
Artificial Intelligence (AI) is currently being introduced into different domains, including medicine. Specifically in radiation oncology, machine learning models allow automation and optimization of the workflow. A lack of knowledge and interpretation of these AI models can hold back wide-spread and full deployment into clinical practice. To facilitate the integration of AI models in the radiotherapy workflow, generally applicable recommendations on implementation and quality assurance (QA) of AI models are presented. For commonly used applications in radiotherapy such as auto-segmentation, automated treatment planning and synthetic computed tomography (sCT) the basic concepts are discussed in depth. Emphasis is put on the commissioning, implementation and case-specific and routine QA of AI models needed for a methodical introduction in clinical practice.


Postsynthesis of h-BN/Graphene Heterostructures Inside a STEM.
Combinations of 2D materials with different physical properties can form heterostructures with modified electrical, mechanical, magnetic, and optical properties. The direct observation of a lateral heterostructure synthesis is reported by epitaxial in-plane graphene growth from the step-edge of hexagonal BN (h-BN) within a scanning transmission electron microscope chamber. Residual hydrocarbon in the chamber is the carbon source. The growth interface between h-BN and graphene is atomically identified as largely N-C bonds. This postgrowth method can form graphene nanoribbons connecting two h-BN domains with different twisting angles, as well as isolated carbon islands with arbitrary shapes embedded in the h-BN layer. The electronic properties of the vertically stacked h-BN/graphene heterostructures are investigated by electron energy-loss spectroscopy (EELS). Low-loss EELS analysis of the dielectric response suggests a robust coupling effect between the graphene and h-BN layers.


Varenicline for smoking cessation.
Varenicline (pronounced va-re-nik-leen) (Champix - Pfizer), a nicotinic receptor partial agonist, is the first medicine of this type licensed for smoking cessation in adults. Launched in the UK in December 2006, the drug accounted for 14% of all prescription items and 22% of total expenditure, for smoking cessation products dispensed in the community in 2007. Marketing materials claim that it has a "unique dual action", a "significantly higher quit rate" than bupropion, and a "favourable safety and tolerability profile". Here we discuss the evidence for varenicline and consider its role in smoking cessation.


Cyclosporine-A induces endoplasmic reticulum stress and influences pro-apoptotic factors in human gingival fibroblasts.
Cyclosporine-A (CsA) induces gingival overgrowth. Cyclosporine's anti-apoptotic activity in human gingival fibroblast is due to desensitization of mitochondrial permeability transition pore (MPTP) and augmentation of anti-apoptotic, Bcl2. Alternative mechanisms of apoptosis exist involving enzymes like calcium-dependent Calpain and signaling events related to apoptosis, like Glycogen synthase kinase 3β (GSK3β) and protein kinase A (PKA). Cyclosporine-A in renal tubular cells induces endoplasmic reticulum stress (ER stress) which has not been explored in gingival overgrowth. Hence, this study was carried out to assess the influence of Cyclosporine-on ER stress and on the alternate anti-apoptotic mechanisms. Human gingival fibroblasts were treated with CsA, and expression of ER stress markers, such as binding immunoglobulin protein and CCAAT/enhancer-binding protein homologous protein (CHOP), MPTP, and expression of Calpain & GSK3β /PKA were estimated. The results showed CsA-added fibroblast significantly increasing the expression of Endoplasmic reticulum stress markers. Contrary to usual ER stress outcome of apoptosis, we observed Cyclosporine's anti-apoptotic action in spite of augmented ER stress markers. We conclude that CsA's independent action on different organelles may alter the conventional outcome of ER stress.


Postural changes in lung volumes in patients with heart failure and Cheyne-Stokes respiration: Relationship with sleep apnea severity.
It has been proposed that the increased severity of sleep apnea frequently observed in heart failure (HF) patients with Cheyne-Stokes respiration (CSR) when sleeping in the supine compared to the lateral position, may be caused by the concomitant reduction in functional residual capacity (FRC). We assessed positional changes in FRC in patients with CSR and investigated the relationship between these changes in the laboratory and corresponding changes in CSR severity during sleep. After a diagnostic polysomnography, 18 HF patients with dominant CSR and an apnea-hypopnea index (AHI)≥15 events/h underwent a standard pulmonary function test in the sitting position. Measurements were repeated in the supine, left lateral and right lateral. The latter two measurements were averaged to obtain a single lateral measurement. The FRC in the seated position was 3.0 ± 0.5 L (85 ± 13% of predicted), decreased to 2.3 ± 0.3 L (-21 ± 8%, p < 0.0001) in the supine position, and increased to 2.8 ± 0.4 L (+21 ± 12%, p < 0.0001) from the supine to the lateral position (-5±8% vs seated, p = 0.013). During sleep, the AHI and the apnea index (AI) decreased from 47 ± 15 events/h to 26 ± 12 events/h (-46 ± 20%, p < 0.0001) and from 29 ± 21 events/h to 12 ± 10 events/h (-61 ± 40%, p < 0.001) from the supine to the lateral position. Changes in the AI were significantly correlated with corresponding changes in FRC (ρ = -0.55, p = 0.032). In patients with HF and CSR, lying in the supine position causes a significant reduction in FRC in the context of a chronically reduced FRC. The negative correlation between postural changes in FRC and AI supports the hypothesis that the reduction in lung gas stores in the supine position may promote/exacerbate respiratory control instability.


Detection and characterization of cyclodextrin complexes with beta-carboline derivatives by spectroscopic techniques.
beta-Carboline alkaloids exhibit a great variety of pharmacological activities. The solid inclusion complexes of harmane and harmine with beta-cyclodextrin and also with hydroxypropyl-beta-cyclodextrin, have been prepared following different procedures. IR and NMR spectroscopies were employed to verify the interaction of the guest molecules with the cyclodextrin cavities. The differences observed in the IR and NMR spectra are in agreement with those described in the literature for other guest molecules. The shifts in the 13C- and 1H-NMR spectra confirm the existence of the inclusion complexes. The fluorescence emission spectra of these complexes dissolved in buffered aqueous solution (pH 7.3) exhibit the characteristic peaks of the cationic form for harmane alkaloids. The neutral bands are not present for the free alkaloids in aqueous solutions. Fluorescence quenching emission of the complexes is compared to that of the corresponding free alkaloids.


Electrically conductive membrane distillation via an alternating current operation for zero liquid discharge.
Membrane distillation (MD) shows promise for achieving high salinity treatment and zero liquid discharge (ZLD) compared to conventional water treatment processes due to its unique characteristics, including low energy consumption and high resulting water quality. However, performance degradation due to fouling and scaling under high recovery conditions remains a challenge, particularly considering the need to control both cations and anions for maximum scaling mitigation. Accordingly, in this study, alternating current (AC) operation for electrically conductive membrane distillation (ECMD) is newly proposed, based on its potential for controlling both cations and anions, in contrast to conventional direct current (DC) operation. Systematic experiments and theoretical analysis show that water recovery in ECMD can be increased by 27% through AC operation. The proposed modification and effective AC operation of ECMD increase the practicality of using MD in desalination for a high recovery rate, perhaps even for ZLD.


Determination of crystal violet in water by direct solid phase spectrophotometry after rotating disk sorptive extraction.
The microextraction of crystal violet (CV) from water samples into polydimethylsiloxane (PDMS) using the rotating disk sorptive extraction (RDSE) technique was performed. The extracting device was a small Teflon disk that had an embedded miniature magnetic stirring bar and a PDMS (560 μL) film attached to one side of the disk using double-sided tape. The extraction involves a preconcentration of CV into the PDMS, where the analyte is then directly quantified using solid phase spectrophotometry at 600 nm. Different chemical and extraction device-related variables were studied to achieve the best sensitivity for the determination. The optimum extraction was performed at pH 14 because under this condition, CV is transformed to the neutral and colorless species carbinol, which can be quantitatively transferred to the PDMS phase. Although the colorless species is the chemical form extracted in the PDMS, an intense violet coloration appeared in the phase because the -OH bond in the carbinol molecule is weakened through the formation of hydrogen bonds with the oxygen atoms of the PDMS, allowing the resonance between the three benzene rings to compensate for the charge deficit on the central carbon atom of the molecule. The accuracy and precision of the method were evaluated in river water samples spiked with 10 and 30 μg L(-1) of CV, yielding a relative standard deviation of 6.2% and 8.4% and a recovery of 98.4% and 99.4%, respectively. The method detection limit was 1.8 μg L(-1) and the limit of quantification was 5.4 μg L(-1), which can be decreased if the sample volume is increased.


Seminoma and parathyroid adenoma in a snow leopard (Panthera unica).
A seminoma and parathyroid adenoma were diagnosed in an aged snow leopard. The ultrastructural appearance of the seminoma was similar to that described in the dog and in man. The lack of significant amounts of rough endoplasmic reticulum, Golgi complexes and free ribosomes in the parathyroid adenoma suggested that it was non-functional. Parathyroid adenoma has not been previously described in a large wild feline.


Inhibition of MLC20 phosphorylation downstream of Ca2+ and RhoA: A novel mechanism involving phosphorylation of myosin phosphatase interacting protein (M-RIP) by PKG and stimulation of MLC phosphatase activity.
Previous studies have shown that cGMP-dependent protein kinase (PKG) act on several targets in the contractile pathway to reduce intracellular Ca(2+) and/or augment RhoA-regulated myosin light chain phosphatase (MLCP) activity and cause muscle relaxation. Recent studies have identified a novel protein M-RIP that associates with MYPT1, the regulatory subunit of MLCP. Herein, we examine whether PKG enhance MLCP activity downstream of Ca(2+) and RhoA via phosphorylation of M-RIP in gastric smooth muscle cells. Treatment of permeabilized muscle cells with 10 μM Ca(2+) caused an increase in MLC20 phosphorylation and muscle contraction, but had no effect on Rho kinase activity. Activators of PKG (GSNO or cGMP) decreased MLC20 phosphorylation and contraction in response to 10 μM Ca(2+), implying existence of inhibitory mechanism independent of Ca(2+) and RhoA. The effect of PKG on Ca(2+)-induced MLC20 phosphorylation was attenuated by M-RIP siRNA. Both GSNO and 8-pCPT-cGMP induced phosphorylation of M-RIP; phosphorylation was accompanied by an increase in the association of M-RIP with MYPT1 and MLCP activity. Taken together, these results provide evidence that PKG induces phosphorylation of M-RIP and enhances its association with MYPT1 to augment MLCP activity and MLC20 dephosphorylation and inhibits muscle contraction, downstream of Ca(2+)- or RhoA-dependent pathways.


Effect of seminal leukocytes on in vitro fertilization and intracytoplasmic sperm injection outcomes.
To investigate the influence of seminal leukocytes on conventional IVF and intracytoplasmic sperm injection (ICSI) outcomes, using a flow cytometry method. Prospective study. Tertiary infertility center and research institute. One hundred sixty-four couples undergoing conventional IVF or ICSI. Seminal leukocytes were counted by flow cytometry. Correlation between seminal leukocytes concentration and reproductive outcomes in IVF and ICSI cycles. The median number of oocytes retrieved, the fertilization and cleavage rate, the median number and grade of embryos transferred, the median number of good-quality embryos transferred, and the median percentage of good-quality embryos from total embryos transferred, in leukocytospermic and non-leukocytospermic patients were not statistically different after either IVF or ICSI. Similarly, there were no significant differences between the two groups for implantation rate and clinical pregnancy rate. Multivariate logistic regression analysis showed that the reproductive outcomes were not influenced by adjustment for female age, infertility diagnosis, number of previous attempts, treatment protocol (GnRH agonist or antagonist), assisted reproduction procedure (IVF or ICSI), and leukocytospermia. By profiling the proper Poisson regression models, no leukocytospermia cut-off value was able to identify the subjects at risk for oocyte fertilization or embryo cleavage failure. Using a flow cytometry method, we demonstrated that leukocytospermia does not significantly influence IVF or ICSI outcomes. The same results were obtained by using lower or higher cut-off values for leukocytospermia (from 0.2 to 2 × 10(6)/mL).


The regulatory roles of phosphatases in cancer.
The relevance of potentially reversible post-translational modifications required for controlling cellular processes in cancer is one of the most thriving arenas of cellular and molecular biology. Any alteration in the balanced equilibrium between kinases and phosphatases may result in development and progression of various diseases, including different types of cancer, though phosphatases are relatively under-studied. Loss of phosphatases such as PTEN (phosphatase and tensin homologue deleted on chromosome 10), a known tumour suppressor, across tumour types lends credence to the development of phosphatidylinositol 3-kinase inhibitors alongside the use of phosphatase expression as a biomarker, though phase 3 trial data are lacking. In this review, we give an updated report on phosphatase dysregulation linked to organ-specific malignancies.


Behavior of a GPI-anchored protein in phospholipid monolayers at the air-water interface.
The interaction between alkaline phosphatase (AP), a glycosylphosphatidylinositol (GPI)-anchored protein (AP-GPI), and phospholipids was monitored using Langmuir isotherms and PM-IRRAS spectroscopy. AP-GPI was injected under C16 phospholipid monolayers with either a neutral polar head (1,2-dipalmitoyl-sn-glycero-3-phosphocholine monohydrate (DPPC)) or an anionic polar head (1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS)). The increase in molecular area due to the injection of protein depended on the surface pressure and the type of phospholipid. At all surface pressures, it was highest in the case of DPPS monolayers. The surface elasticity coefficient E, determined from the pi-A diagrams, allowed to deduct that the AP-GPI-phospholipid mixtures presented a molecular arrangement less condensed than the corresponding pure phospholipid films. PM-IRRAS spectra suggested different protein-lipid interactions as a function of the nature of the lipids. AP-GPI modified the organization of the DPPS deuterated chains whereas AP-GPI affected only the polar group of DPPC at low surface pressure (8 mN/m). Different protein hydration layers between the DPPC and DPPS monolayers were suggested to explain these results. PM-IRRAS spectra of AP-GPI in the presence of lipids showed a shape similar to those collected for pure AP-GPI, indicating a similar orientation of AP-GPI in the presence or absence of phospholipids, where the active sites of the enzyme are turned outside of the membrane.


Norethisterone Related Drug Induced Liver Injury: A Series of 3 Cases.
Drug induced liver injury (DILI) is uncommon and severe forms are associated with significant morbidity and mortality. Female sex hormones (estrogens and progestogens) related DILI generally occur with estrogen component. Progesterone component related DILI are infrequently reported. Norethisterone is commonly used drug in gynecologic practice to prevent excess per vaginal bleeding. We report 3 cases of Norethisterone related DILI manifesting as significant rise of transaminases. All of these patients took Norethisterone for prolonged periods and improved completely after withdrawal of drug.


Assessing the size and stage of testicular germ cell tumours: 1984-2003.
To assess the size and stage of testicular tumours on presentation in the period 1984-2002. Demographic details and information on staging on 550 patients treated at St. Bartholomew's and the Royal London Hospital in the period 1984-2002 were collected prospectively in the departmental database. Information on testicular size was obtained by reviewing the histopathology records, and the maximum dimension of the tumour as measured in the gross specimen was taken as the size of the testicular tumour. The period 1984-2002 was divided into three intervals, i.e. 1984-95, 1996-98 and 1999-2002. The mean testicular tumour size in the three intervals decreased from 4 cm (162 tumours) to 3.2 cm (85) and 2.5 cm (72; P = 0.002, Student's t-test). The proportion of tumours of <2 cm on presentation also increased, from 11% to 14% and 23% in the three intervals, respectively, while the proportion of patients with stage 1 disease increased from 57%, to 63% and 77%, respectively. The size of testicular tumours on presentation has shown a consistent decline in the last two decades, the mean size now being 2.5 cm. That 23% are now <2 cm raises the possibility of testis-preserving surgery in this young group of patients, who have an excellent prognosis, and therefore in the long-term issues such as psychological morbidity and natural fertility assume greater importance. There is a need for a randomized controlled trial to evaluate these issues.


Posterior vitreous detachment with plasmin in the isolated human eye.
To investigate the efficacy and safety of intravitreal injection of plasmin in producing posterior vitreous detachment (PVD), and the possible role of plasmin in degrading adhesion glycoproteins of the internal limiting membrane (ILM). A total of 20 young human cadaver eyes were randomized into four groups. Groups 1, 2, and 3 were injected with 1, 2, and 3 U of human plasmin, respectively, and group 1 was injected with 0.1 ml balanced salt solution as control. Optical and transmission electron microscopy were used to examine the ultrastructure of the vitreoretinal interface. The amounts of fibronectin (FN) and laminin (LN) in the ILM were determined using electron-immunocytochemical techniques. The survival of retinal cells was evaluated using flow cytometry. Analysis of variance (ANOVA) and the Tukey test were used for statistical analysis. Microscopy showed that plasmin, especially in group 4, cleaved the attachment of the vitreous collagen fibrils to the ILM. There was no evidence of damage to the inner retina except some post-mortem lytic changes. The amounts of LN and FN in the ILM were decreased following injection of plasmin. There were statistically significant differences in the amounts of FN in groups 3 and 4 compared to the control group (P<0.05), and in the amount of LN in group 4 (P<0.05). Retinal cell viability was similar in both treated and control eyes. Human plasmin can disrupt the attachment of the posterior hyaloid to the ILM without producing morphological changes or acute toxicity of the inner retina of the human eye. PVD can be induced completely by injection of 3 U plasmin. The enzyme may be useful for removing the cortical vitreous during vitreous surgery.


Quantifying impacts of human pressures on ecosystem services: Effects of widespread non-indigenous species in the Baltic Sea.
Ecosystem services (ES) are the benefits natural ecosystems provide to society, such as food provisioning, water supply, climate regulation and recreational benefits. Biological invasions are a major driver of global change, and several non-indigenous species (NIS) may alter key ecological feedbacks with ultimate consequences to ES, livelihoods and human wellbeing. Nonetheless, the effects of NIS on ES supply remain largely unquantified. Here we present the first quantitative case study assessing the impacts of widespread NIS on ES in the Baltic Sea, by developing and employing a robust and repeatable data-driven approach. All NIS with a sufficient knowledge base pose large and highly significant effects on ES, resulting on average 55 % change in the intensity of ES. Most impacts affected regulation services, concerning both abiotic and biotic realms, with little evidence on cultural and provisioning services. The methodology can be easily employed beyond the current study realm e.g. to better understand the roles of human pressures on ES in any ecosystem. Importantly, the study also identified major biases not only in the availability of taxonomic and sub-regional evidence, but also in the different study types employed to create the evidence base.


Disappearance of soft drusen and subsequent development of choroidal neovascularization following macular hole surgery: a case report.
Drusen are important risk factor for neovascular age-related macular degeneration (AMD) and have a dynamic nature as they can enlarge, newly form, or disappear over time. There have been few reports on drusen regression or choroidal neovascularization (CNV) development after macular hole surgery. We report, to our knowledge, the first case of both drusen regression and subsequent CNV development within 7 months of successful macular hole surgery. A 73-year-old woman presented with a stage 3 full-thickness macular hole and large, confluent soft macular drusen in the right eye and a neovascular age-related macular degeneration (AMD) in the fellow eye. Four months after the successful macular hole surgery, significant regression of drusen was seen, especially in the temporal area to the fovea. Three months later, the patient developed CNV and her best-corrected visual acuity decreased to 20/100, despite further regression of macular drusen. Macular hole patients with macular soft drusen need to be carefully followed up after surgery for possible drusen regression and CNV development.


Identification of marker genes to monitor residual iPSCs in iPSC-derived products.
Engineered tissues and cell therapies based on human induced pluripotent stem cells (iPSCs) represent a promising approach for novel medicines. However, iPSC-derived cells and tissues may contain residual undifferentiated iPSCs that could lead to teratoma formation after implantation into patients. As a consequence, highly sensitive and specific methods for detecting residual undifferentiated iPSCs are indispensable for safety evaluations of iPSC-based therapies. The present study provides an approach for identifying potential marker genes for iPSC impurities in iPSC-derived cells using RNA sequencing data from iPSCs and various differentiated cell types. Identifying iPSC marker genes for each cell type individually provided a larger and more specific set of potential marker genes than considering all cell types in the analysis. Thus, the authors focused on identifying markers for iPSC impurities in iPSC-derived cardiomyocytes (iCMs) and validated the selected genes by reverse transcription quantitative polymerase chain reaction. The sensitivity of the candidate genes was determined by spiking different amounts of iPSCs into iCMs and their performance was compared with the previously suggested marker lin-28 homolog A (LIN28A). Embryonic stem cell-related gene (ESRG), long intergenic non-protein coding RNA 678 (LINC00678), CaM kinase-like vesicle-associated (CAMKV), indoleamine 2,3-dioxygenase 1 (IDO1), chondromodulin (CNMD), LINE1-type transposase domain containing 1 (L1DT1), LIN28A, lymphocyte-specific protein tyrosine kinase (LCK), vertebrae development-associated (VRTN) and zinc finger and SCAN domain containing 10 (ZSCAN10) detected contaminant iPSCs among iCMs with a limit of detection that ranged from 0.001% to 0.1% depending on the gene and iCM batch used. Using the example of iCMs, the authors provide a strategy for identifying a set of highly specific and sensitive markers that can be used for quality assessment of iPSC-derived products.


Pulmonary tumor with Notochordal differentiation: a case report and morphologic, Immunohistochemical and molecular study of benign Notochordal cell tumor originating in the lung.
Extraosseous benign notochordal cell tumor is extremely rare, and there are only five reported cases worldwide. The presented case of pulmonary primary benign notochordal cell tumor is the sixth case, but the first to report the deletion mutation of EGFR gene exon 19. The patient was a 50-year-old asymptomatic woman, who had been followed up for 3 years for a nodule in the right lung. After ten months of the wedge resection, the patient is alive without evidence of recurrence or metastasis. The tumor was 7 mm in diameter and was well demarcated. The tumor was consisted of a sheet of large round vacuolated cells with small and bland nuclei. No connective tissue containing blood vessels or inflammatory cell infiltration was detected in the stroma. The tumor was positive for CK AE1/AE3, Vimentin, S100 and Brachyury. EGFR gene mutation and amplification were not detected. We firstly reported the positive immunohistochemical staining for EGFR and the negative molecular results of EGFR gene of pulmonary primary benign notochordal cell tumor. Due to the rarity of this tumor, more reports are needed to explore pathological characteristics, especially the molecular characteristics, in order to better understand the nature of tumors.


Cannabinoid receptor 1 contributes to sprouted innervation in endometrial ectopic growth through mitogen-activated protein kinase activation.
The endocannabinoid system regulates neurite outgrowth and neurogenesis during development of the central nervous system. Cannabinoid receptor 1 (CB1R) is expressed in neurons, including the somata and fibers, that innervate the endometrial ectopic cyst in rats. Here, we investigated the contribution of CB1R and its downstream signaling to the innervation of endometrial ectopic growth. We found that intrathecal injection of a CB1R agonist enhanced both the density of protein gene product (PGP) 9.5-immunoreactive sprouted nerve fibers and the protein level of PGP 9.5 of the ectopic cyst, and the CB1R antagonist induced opposite effects. The CB1R agonist increased the expression of phosphorylated extracellular signal-regulated kinase (pERK) and c-Jun N-terminal kinase (pJNK), but not pp38, in dorsal root ganglion (DRG), whereas the CB1R antagonist only decreased the expression of pERK. In cultured DRG neurons, CB1R agonists dose-dependently increased neurite elongation. The mitogen-activated protein kinase (MAPK)/ERK kinase (MEK) and JNK inhibitors, but not the p38 inhibitor, attenuated CB1R agonist-induced neurite elongation. The inhibitions of CB1R and its downstream ERK and JNK signaling pathways may alleviate the sprouted innervation that has been involved in ENDO-associated pain. This finding may provide a new therapeutic target for patients with endometriosis.


S100A8 and S100A9 in inflammation and cancer.
Calprotectin (S100A8/A9), a heterodimer of the two calcium-binding proteins S100A8 and S100A9, was originally discovered as immunogenic protein expressed and secreted by neutrophils. Subsequently, it has emerged as important pro-inflammatory mediator in acute and chronic inflammation. More recently, increased S100A8 and S100A9 levels were also detected in various human cancers, presenting abundant expression in neoplastic tumor cells as well as infiltrating immune cells. Although, many possible functions have been proposed for S100A8/A9, its biological role still remains to be defined. Altogether, its expression and potential cytokine-like function in inflammation and in cancer suggests that S100A8/A9 may play a key role in inflammation-associated cancer.


Double tumours of the liver following intravenous thorotrast injection. Patho-anatomical report on two cases.
Observations on the contrast between the livers of two patients who underwent an arteriogram with thorotrast in 1941 have been made. The first case produced a cholangicellular carcinoma (CCC) in addition to atypical proliferations of sinusoidal lining cells. The second case produced a very rare tumour combination of a CCC with a haemangioendothelial sarcoma (Haem-SA the problem of double tumours of the liver following thorotrast injection is discussed in the light of these cases. In addition the discussion includes various opinions on Kupffer cells.


Fast and Cost-Efficient 17 O-Isotopic Labeling of Carboxylic Groups in Biomolecules: From Free Amino Acids to Peptide Chains.
17 O NMR spectroscopy is a powerful technique, which can provide unique information regarding the structure and reactivity of biomolecules. However, the low natural abundance of 17 O (0.04 %) generally requires working with enriched samples, which are not easily accessible. Here, we present simple, fast and cost-efficient 17 O-enrichment strategies for amino acids and peptides by using mechanochemistry. First, five unprotected amino acids were enriched under ambient conditions, consuming only microliter amounts of costly labeled water, and producing pure molecules with enrichment levels up to ∼40 %, yields ∼60-85 %, and no loss of optical purity. Subsequently, 17 O-enriched Fmoc/tBu-protected amino acids were produced on a 1 g/day scale with high enrichment levels. Lastly, a site-selective 17 O-labeling of carboxylic functions in peptide side-chains was achieved for RGD and GRGDS peptides, with ∼28 % enrichment level. For all molecules, 17 O ssNMR spectra were recorded at 14.1 T in reasonable times, making this an important step forward for future NMR studies of biomolecules.


An investigation of the effect of brain atrophy on brain injury in multiple sclerosis.
Multiple sclerosis (MS) is a disease of the central nervous system (CNS) that affects the brain and spinal cord. It is estimated that the average prevalence of MS is 35.9 cases per 100,000 and a total of 2.8 million people worldwide have MS. Brain atrophy is usually seen in the early stages of MS, and its progress is faster than healthy people. The present study was a numerical study that uses the Fluid-structure interaction (FSI) model to investigate the effect of brain atrophy on brain injury in MS. Firstly, a healthy model was constructed from MRI images and validated by experimental data. Then three models with different degrees of brain atrophy, which showed the rate of brain atrophy in different years in MS patients, were developed to model the brain atrophy in MS. The models were subjected to two different types of impact conditions. Type I, which only produced a translational motion and the HIC value of 744, was applied to each model. Type II produced both translational and rotational motion. In this type of impact, the experimental kinematics, with peaks of 450 g for the translational acceleration and 26.2 krad/s2 for the rotational acceleration, were applied to the nodes that located in the center of gravity of the head models and the results were extracted from each one. According to the results of impact type I, the pressure of the frontal lobe of the brain is 149,647 Pa in the health model and 137,690 Pa in the model with severe atrophy.


Traces of black carbon sources before and after the Covid-19 outbreak in Tehran, Iran.
The concentration of black carbon was measured in four sites of the industrial and high-traffic metropolis of Tehran with different land uses. Then, the contribution of biomass and fossil fuels in the emission of this pollutant was modeled using the Aethalometer model. The possible locations of important sources of black carbon dissemination were projected using PSCF and CWT models, and the results were compared in the two periods before and after the Covid-19 outbreak. Temporal variations of black carbon illustrated that BC concentration decreased in the period after the onset of the pandemic in all studied areas, and this decline was more explicit in the traffic intersection of the city. Diurnal changes of BC concentration indicated the significant impact of the application of the law banning night traffic of motor vehicles in reducing the BC concentration in this period, and probably the reduction of HDDV traffic has played the most important role in this reduction. The results related to the share of BC sources indicated that black carbon emissions are affected by an average of about 80% of fossil fuel combustion and wood combustion interferes with about 20% of BC emissions. Finally, speculations were made about the possible sources of BC emission and its urban scale transport using PSCF and CWT models, which indicated the superiority of the CWT model in terms of source segregation. The results of this analysis were further utilized to surmise black carbon emission sources based on the land use of receptor points.


Changes in EEG, blood levels, mood scales and performance scores during long term treatment with diazepam, phenobarbital or placebo in patients.
1. The patient population consisting of fifteen patients was divided into three groups, namely: diazepam group, phenobarbital group and placebo group. After three weeks the medicated groups were switched to placebo for a week and the placebo group was given phenobarbital. 2. The parameters to be assessed once a week comprised frequency-analyzed EEG recordings, performance in two attention tests and subjectively estimated mood modalities. 3. The EEG analysis suggested that EEG patterns: a) were drug-dependent, with a differential distribution for each drug of the four frequency bands analyzed; b) showed no change during the three-week treatment period; c) changed on cessation of medication or on switch from placebo to active medication; d) were task-dependent and changed in a systematic way with the level of activation (stress, vigilance or relaxation). 4. The results would allow a better understanding of the clinical course, the choice of therapeutic measures and of the underlying mechanisms of action.


Incidence and risk of ptosis following ocular surgery: a systematic review and meta-analysis.
We conducted a systematic review and meta-analysis to evaluate the incidence and risk factors of ptosis following ocular surgery. PubMed, Embase, and Cochrane Library were searched for articles that assessed the incidence or risk factors of ptosis following ocular surgery up to October 2017. We used a fixed effects model to calculate a pooled estimate of incidence, with subgroup analyses to evaluate the effect of different variables. The relative risks (RRs) or odds ratios (ORs) and 95% confidence intervals (CIs) for all available factors were calculated using the fixed effects models. A total of 16 studies on 2856 eyes were analyzed, including 3 randomized controlled trials (RCTs) and 13 cohort studies. The overall incidence of ptosis following ocular surgery was 11.4% (95% CI 10.1-12.8%). Subgroup analyses showed that the region and the surgery type were significantly associated with the incidence of postoperative ptosis. Men were less likely to get postoperative ptosis than women (OR 0.62; 95% CI 0.43-0.89). However, age (OR 0.77; 95% CI 0.48-1.23), side (OR 1.37; 95% CI 0.84-2.25), type of anesthesia (OR 0.57; 95% CI 0.16-2.05), prior surgery (OR 1.09; 95% CI 0.64-1.83), bridle suture (OR 2.04; 95% CI 0.94-4.42), or combined surgery (OR 0.95; 95% CI 0.58-1.57) did not significantly change the risk of ptosis following ocular surgery. More than one in ten patients who undergo ocular surgery will develop ptosis. Different regions and surgery types may influence the occurrence of this abnormality. Female gender is a risk factor for development of postoperative ptosis.


β-Tubulin mutations that cause severe neuropathies disrupt axonal transport.
Microtubules are fundamental to neuronal morphogenesis and function. Mutations in tubulin, the major constituent of microtubules, result in neuronal diseases. Here, we have analysed β-tubulin mutations that cause neuronal diseases and we have identified mutations that strongly inhibit axonal transport of vesicles and mitochondria. These mutations are in the H12 helix of β-tubulin and change the negative charge on the surface of the microtubule. This surface is the interface between microtubules and kinesin superfamily motor proteins (KIF). The binding of axonal transport KIFs to microtubules is dominant negatively disrupted by these mutations, which alters the localization of KIFs in neurons and inhibits axon elongation in vivo. In humans, these mutations induce broad neurological symptoms, such as loss of axons in the central nervous system and peripheral neuropathy. Thus, our data identified the critical region of β-tubulin required for axonal transport and suggest a molecular mechanism for human neuronal diseases caused by tubulin mutations.


Identification of α-Tocopherol succinate as an RFFL-substrate interaction inhibitor inducing peripheral CFTR stabilization and apoptosis.
The E3 ubiquitin ligase RFFL is an apoptotic inhibitor highly expressed in cancers and its knockdown suppresses cancer cell growth and sensitizes to chemotherapy. RFFL also participates in peripheral protein quality control which removes the functional cell surface ΔF508-CFTR channel and reduces the efficacy of pharmaceutical therapy for cystic fibrosis (CF). Although RFFL inhibitors have therapeutic potential for both cancer and CF, they remain undiscovered. Here, a chemical array screening has identified α-tocopherol succinate (αTOS) as an RFFL ligand. NMR analysis revealed that αTOS directly binds to RFFL's substrate-binding region without affecting the E3 enzymatic activity. Consequently, αTOS inhibits the RFFL-substrate interaction, ΔF508-CFTR ubiquitination and elimination from the plasma membrane of epithelial cells, resulting in the increased functional CFTR channel. Among the α-tocopherol (αTOL) analogs we tested, only αTOS inhibited the RFFL-substrate interaction and increased the cell surface ΔF508-CFTR, depending on RFFL expression. Similarly, the unique proapoptotic effect of αTOS was dependent on RFFL expression. Thus, unlike other αTOL analogs, αTOS acts as an RFFL protein-protein interaction inhibitor which may explain its unique biological properties among αTOL analogs. Moreover, αTOS may act as a CFTR stabilizer, a novel class of drugs that extend cell surface ΔF508-CFTR lifetime.


Maternal health in pregnancy and intellectual disability in the offspring: a population-based study.
The aim of the study is to investigate the relationship between common maternal conditions and intellectual disability (ID) of unknown cause in the offspring. Information about the maternal health of children with and without ID was obtained by using record linkage. For mothers with specific medical conditions, proportions of children with mild to moderate ID, severe ID, and autism spectrum disorder (ASD) with ID were compared with those who did not have ID. There was an increased risk for mild to moderate ID in children of mothers with asthma (odds ratio [OR], 1.52; confidence interval [CI], 1.26-1.83]), diabetes (OR, 1.69; CI, 1.26-2.27), a renal or urinary condition (OR, 2.09; CI, 1.39-3.14), and epilepsy (OR, 3.53; CI, 2.56-4.84). ASD risk was increased for children of women with diabetes (OR, 2.89; CI, 1.28-6.51) and epilepsy (OR, 4.57; CI, 1.69-12.31). For anemia (n = 1101), there was an increased risk for severe ID (OR, 5.26; CI, 2.16-12.80). The increased risk for ID in offspring of mothers with such conditions as asthma and diabetes is particularly important for disadvantaged or ethnic populations, for whom these conditions are more prevalent and may be less well managed.


A synergetic theory of environmentally-specified and learned patterns of movement coordination. I. Relative phase dynamics.
This paper outlines and applies a synergetic strategy to the coordination of human rhythmical movement. It extends earlier empirical and theoretical work to include the influence of specific environmental information and of memory on the dynamics of the collective variables (order parameters) that characterize the coordination patterns. Key ideas concern cooperative and competitive influences on the collective dynamics. Recent experiments on environmentally specified and learned rhythmic movement patterns are modeled explicitly on the level of the collective variable, relative phase. New predictions are presented and research directions proposed that follow directly from the present theoretical approach.


Rapid replacement of prevailing genotype of human respiratory syncytial virus by genotype ON1 in Beijing, 2012-2014.
Human respiratory syncytial virus (HRSV) is the most common viral pathogen causing lower respiratory infections in infants and young children worldwide. HRSV ON1 genotype in subgroup A with a characteristic of a 72 nucleotide duplication in the second highly variable region of attachment glycoprotein gene, has been reported in some countries since it was first detected in clinical samples collected in Canada in 2010. In this study, 557 HRSV antigen-positive nasopharyngeal aspirates were randomly selected during 2012/2013 to 2013/2014 HRSV seasons in Beijing for subgroup typing and for ON1 genotype screening by using a PCR based method developed for easily identifying genotype ON1 out of strains of subtype A. It was found that subgroup B was dominant in the 2012/2013 season and sudden shift of subgroup dominance from B to A and rapid replacement of previously prevailing NA1 genotype by ON1 genotype occurred in the 2013/2014 season. Reversible amino acid replacement in the G protein gene was found in a new branch of ON1 genotype. The evolutionary rate of the 351 global ON1 sequences was estimated to 7.34 × 10(-3) nucleotide substitutions per site per year (95% highest probability density intervals, HPD, 5.71 × 10(-3) to 9.04 × 10(-3)), with the time of most recent common ancestor dating back to June 2009.


High incidence of cervical human papillomavirus infection in women during their first sexual relationship.
The prevalence of cervical human papillomavirus increases with increasing numbers of sexual partners, leaving the impression that this infection is acquired only as a result of high risk sexual behaviour. Using longitudinal data from 242 women who had only had one sexual partner, we found that the risk of acquiring cervical human papillomavirus infection was 46% (95% CI 28-64) at three years after first intercourse and that the median time from first intercourse to first detection of human papillomavirus was only three months.


Cyclic-AMP response element-based signaling assays for characterization of Trk family tyrosine kinases modulators.
Neurotrophins (NTs) induce gene transcription by binding their high-affinity tropomyosin-related kinase (Trk) receptors and initiating intracellular signal transduction cascades. In particular, activation of the cyclic AMP response element (CRE) in the promoters of target genes serves as surrogate markers for Trk receptor activation as demonstrated in both in vivo and in vitro systems. We used a HEK293 cell line stably expressing a CRE-luciferase reporter gene to develop an assay for monitoring Trk activation in response to their cognate ligands. Using TrkB, we showed that the assay was sensitive to physiological concentrations of brain-derived neurotrophic factor (BDNF) and that the signal was sufficiently robust to be suitable for implementation in high-throughput format. Further characterization of the TrkB expressing stable cell lines showed high-affinity binding for BDNF, a high density of receptor expression, and supported BDNF-mediated phosphorylation signaling. Consistent with this, inhibitors of phosphatidylinositol 3-kinase and the phospholipase C-gamma pathways led to reduction of BDNF-mediated luciferase responses. In contrast, inhibitors of mitogen-activated protein kinase pathways further potentiated BDNF responses. This assay was NT-Trk receptor pair-selective and shown to be further applicable to other Trk family members. This assay may be useful in screening compound libraries to identify Trk agonists, which may be applied towards discriminating between the activities of the different Trk receptor family members and the development of pharmacological drugs.


Senile myoclonic epilepsy of Genton: two cases in Down syndrome with dementia and late onset epilepsy.
Senile myoclonic epilepsy of Genton is a newer epileptic syndrome of the older patients with Down syndrome associated with an Alzheimer-type dementia. We report two observations in whom the clinical and electroencephalographic features are consistent with the description of this syndrome. Both experienced a progressive deterioration of cognitive functions few years before the onset of the epilepsy. The EEG was characteristic with generalized fast spike-waves or polyspikes or polyspike-waves with or without bilateral myoclonic jerks especially at awakenings. One patient had a photoparoxysmal response (11-21Hz) with bilateral myoclonic jerks. Hence, the senile myoclonic epilepsy of Genton is an easily recognizable newer epileptic syndrome of the older Down syndrome patient.


Structural analysis of polymers of sickle cell hemoglobin. II. Sickle hemoglobin macrofibers.
Sickle cell hemoglobin macrofibers are an important intermediate in the low pH crystallization pathway of deoxygenated hemoglobin S that link the fiber to the crystal. Macrofibers are a class of helical particles differing primarily in their diameters but are related by a common packing of their constituent subunits. We have performed three-dimensional reconstructions of three types of macrofibers. These reconstructions show that macrofibers are composed of rows of Wishner-Love double strands in an arrangement similar to that in the crystal. We have measured the orientation and co-ordinates of double strands in macrofibers using cross-correlation techniques. In this approach, the electron density projections of double strands calculated from the known high-resolution crystal structure are compared with regions along the length of the particles in which the distinct pattern of double strands in c-axis projection may be observed. Contrary to assertions by Makinen & Sigountos (1984), our results unambigously demonstrate that adjacent rows of double strands in macrofibers are oriented in an antiparallel manner, as in the Wishner-Love crystal. Adjacent rows of antiparallel double strands are displaced along the helical axis relative to their co-ordinates in the crystal. Electron density models of macrofibers based on the crystallographic structure of the sickle hemoglobin double strand are in good agreement with the projections of macrofibers observed in electron micrographs. We have studied the structure of a closely related crystallization intermediate, the sickle hemoglobin paracrystal. The arrangement of double strands in paracrystals is similar to that in Wishner-Love crystals, except that they are displaced along the a-axis of the crystal. Measurements of the double strand co-ordinates reveal that the distribution of strand positions is bimodal. These results further establish the close structural relationship between macrofibers and paracrystals as intermediates in the crystallization of deoxygenated sickle hemoglobin.


Phase transition and hardness evolution of a Ti-5Al-5Mo-1Fe-1Cr alloy subjected to isothermal aging.
In this paper, a Ti-5Al-5Mo-1Fe-1Cr alloy was isothermally aged at 500 °C for different periods. Phase transformation took place during the aging treatment. The phase transformation was studied by X-ray diffraction spectroscopy and transmission electron microscopy. The as-quenched sample was composed of matrix β (body-centered cubic structure) phase, ωath (hexagonal structure) phase and minor α (hexagonal close-packed structure) phase. The results showed that aging treatment promoted the β → ω → α phase transformations in the as-quenched alloys. Different isothermal aging time affected the contents and morphologies of the three phases inside the alloy and thus the hardness. It was found that the content of hard ω phase plays an important role in the hardness of the Ti-5Al-5Mo-1Fe-1Cr alloy. The hardness of alloy showed an overall decreasing trend as the aging time increased to 2 h, except for the temporary slight increase of the hardness at an aging time of 1 h.


Abnormal eye movements in Parkinson's disease: From experimental study to clinical application.
Parkinson's disease (PD) is a neurodegenerative disease that integrates a series of motor symptoms and non-motor symptoms, making early recognition challenging. The exploration of biomarkers is urgently required. Abnormal eye movements in PD have been reported to appear in a variety of ways since eye tracking technology was developed, such as decreased saccade amplitude, extended saccade latency, and unique saccade patterns. Non-invasive, objective and simple eye tracking has the potential to provide effective biomarkers for the PD diagnosis, progression and cognitive impairment, as well as ideas for research into the occurrence and treatment strategy of motor symptoms. In this review, we introduced the fundamental eye movement patterns and typical eye movement paradigms (such as fixation, pro-saccade, anti-saccade, smooth tracking, and visual search), summarized the symptoms of various ocular motor abnormalities in PD, and discussed the research implications of oculomotor investigation to the pathogenesis of PD and related motor symptoms, as well as the clinical implications as biomarkers and its inspiration on treatment.


Traumatic extradural hematoma in the middle cranial fossa base: clinical analysis of 14 cases.
To explore clinical characteristics, CT manifestation and operative indications of traumatic extradural hematoma in the middle cranial fossa base (EHMCFB). A total of 14 cases with traumatic EHMCFB from January 1997 to May 2000 were retrospectively analyzed. Of 14 cases, 12 cases (85.7%) maintained form chan ge of cisterna ambiens for being pressed; 11 cases who received operations were all cured; 3 received conservative therapy, of whom 2 became better and 1 died. In traumatic EHMCFB, the form change of cisterna ambiens occurs easily. Therefore, if there appears progressively aggravated form change of cisterna ambiens after the cisterna ambiens being pressed, an early a nd active operation is necessary and crucial.


Control of secretory granule interaction and exocytosis in pancreatic cells.
Application of the laser-based technique of photon correlation spectroscopy to an in vitro study of the ionic stability and interaction kinetics of zymogen granules isolated from rat exocrine pancreas is described here. In addition the separation from pancreatic acinar cell cytosol of a factor which stabilizes isolated zymogen granules and inhibits cation-induced granule aggregation is outlined. The basis of this action and the significance of the cytosolic inhibitory factor in the regulation of granule mobility and exocytosis in vivo is discussed.


miR-320a regulates high mobility group box 1 expression and inhibits invasion and metastasis in hepatocellular carcinoma.
Several studies have shown that miR-320a induces apoptosis, inhibits cell proliferation, and affects cell cycle progression as a tumour suppressor in many cancers. However, the involvement of miR-320a in the invasion and metastasis of hepatocellular carcinoma (HCC) is still unknown. Endogenous miR-320a and high mobility group box 1 (HMGB1) expressions were assayed by real-time PCR. Luciferase activities were measured using a dual-luciferase reporter assay system. Western blots were used to determine the protein expressions of HMGB1, MMP2, and MMP9. Invasion and metastasis of tumour cells were, respectively, evaluated by the transwell invasion assay and the wound healing assay. The expression of miR-320a was significantly decreased in 24 of 32 (75%) HCC tissues and associated with the invasion and metastasis of HCC. Furthermore, we demonstrated that HMGB1 was a direct target of miR-320a and there was a significant negative correlation between miR-320a and HMGB1 expression in HCC. Ectopic expression or inhibition of miR-320a potently regulated the invasion and metastasis of HCC cells in HMGB1-dependent manner. Our results showed that miR-320a was involved in the invasion and metastasis by targeting HMGB1 and had an anti-metastasis effect in HCC.


Enhanced activity of cefepime-tazobactam (WCK 4282) against KPC-producing Enterobacteriaceae when tested in media supplemented with human serum or sodium chloride.
The aim of this study was to evaluate the in vitro activity of cefepime-tazobactam cation-adjusted Mueller-Hinton broth (CA-MHB) supplemented with 0.85% sodium chloride (NaCl) or 50% human serum in comparison to standard CA-MHB when testing KPC-producing isolates. A total of 209 contemporary Enterobacteriaceae clinical isolates carrying blaKPC were tested, and cefepime-tazobactam (tazobactam at fixed 8mg/L) activity was enhanced 2-fold when tested in CA-MHB supplemented with 0.85% NaCl or 50% human serum (MIC50/90, 8/32mg/L for both media) compared to standard CA-MHB (MIC50/90, 16/64mg/L). Cefepime-tazobactam at a concentration of ≤16mg/L, which is the pharmacokinetics/pharmacodynamics tentative susceptibility breakpoint based on a high dosing regimen of cefepime-tazobactam (2g-2g q8h 90-minute infusion), inhibited 79.4-80.4% of Enterobacteriaceae isolates carrying blaKPC in MHB supplemented with 0.85% NaCl or 50% human serum. A similar decrease in MIC values was observed when cefepime alone was tested against a subset of the isolates (n=54) in CA-MHB supplemented with 50% human serum or 0.85% NaCl; however, imipenem activity against these 54 organisms was similar or 2-fold higher in CA-MHB supplemented with 0.85% of NaCl (MIC50/90, 8/16mg/L) or with 50% human serum (MIC50 and MIC90, 16mg/L) compared standard CA-MHB (MIC50/90, 8/16mg/L). In summary, cefepime-tazobactam MIC values against Enterobacteriaceae isolates carrying blaKPC were consistently lower in media supplemented with human serum or NaCl, which better mimics physiological conditions. These results suggest that this carbapenem-sparing candidate agent has potential to be used to treat infections caused by KPC-producing Enterobacteriaceae.


Cloning vectors for the expression of green fluorescent protein fusion proteins in transgenic plants.
A series of versatile cloning vectors has been constructed that facilitate the expression of protein fusions to the Aequorea victoria green fluorescent protein (GFP) in plant cells. Amino-terminal- and carboxy-terminal protein fusions have been created and visualized by epifluorescence microscopy, both in transgenic Arabidopsis thaliana and after transient expression in onion epidermal cells. Using tandem dimers and other protein fusions to GFP, we found that the previously described localization of wild-type GFP to the cell nucleus is most likely due to diffusion of GFP across the nuclear envelope rather than to a cryptic nuclear localization signal. A fluorescence-based, quantitative assay for nuclear localization signals is described. In addition, we have employed the previously characterized mutants GFP-S65T and GFP-Y66H in order to allow for the expression of red-shifted and blue fluorescent proteins, respectively, which are suitable for double-labeling studies. Expression of GFP-fusions was controlled by a cauliflower mosaic virus 35S promoter. Using the Arabidopsis COP1 protein as a model, we confirmed a close similarity in the subcellular localization of native COP1 and the GFP-tagged COP1 protein. We demonstrated that COP1 was localized to discrete subnuclear particles and further confirmed that fusion to GFP did not compromise the activity of the wild-type COP1 protein.


Effect of Portable Air Filtration Systems on Personal Exposure to Fine Particulate Matter and Blood Pressure Among Residents in a Low-Income Senior Facility: A Randomized Clinical Trial.
Fine particulate matter (smaller than 2.5 μm) (PM2.5) air pollution is a major global risk factor for cardiovascular (CV) morbidity and mortality. Few studies have tested the benefits of portable air filtration systems in urban settings in the United States. To investigate the effectiveness of air filtration at reducing personal exposures to PM2.5 and mitigating related CV health effects among older adults in a typical US urban location. This randomized, double-blind crossover intervention study was conducted from October 21, 2014, through November 4, 2016, in a low-income senior residential building in Detroit, Michigan. Forty nonsmoking older adults were enrolled, with daily CV health outcome and PM2.5 exposure measurements. Participants were exposed to the following three 3-day scenarios separated by 1-week washout periods: unfiltered air (sham filtration), low-efficiency (LE) high-efficiency particulate arrestance (HEPA)-type filtered air, and high-efficiency (HE) true-HEPA filtered air using filtration systems in their bedroom and living room. The primary outcome was brachial blood pressure (BP). Secondary outcomes included aortic hemodynamics, pulse-wave velocity, and heart rate variability. Exposures to PM2.5 were measured in the participants' residences and by personal monitoring. The 40 participants had a mean (SD) age of 67 (8) years (62% men). Personal PM2.5 exposures were significantly reduced by air filtration from a mean (SD) of 15.5 (10.9) μg/m3 with sham filtration to 10.9 (7.4) μg/m3 with LE fitration and 7.4 (3.3) μg/m3 with HE filtration. Compared with sham filtration, any filtration for 3 days decreased brachial systolic and diastolic BP by 3.2 mm Hg (95% CI, -6.1 to -0.2 mm Hg) and 1.5 mm Hg (95% CI, -3.3 to 0.2 mm Hg), respectively. A continuous decrease occurred in systolic and diastolic BP during the 3-day period of LE filtration, with a mean of 3.4 mm Hg (95% CI, -6.8 to -0.1 mm Hg) and 2.2 mm Hg (95% CI, -4.2 to -0.3 mm Hg), respectively. For HE filtration, systolic and diastolic BP decreased by 2.9 mm Hg (95% CI, -6.2 to 0.5 mm Hg) and 0.8 mm Hg (95% CI, -2.8 to 1.2 mm Hg), respectively. Most secondary outcomes were not significantly improved. Results of this study showed that short-term use of portable air filtration systems reduced personal PM2.5 exposures and systolic BP among older adults living in a typical US urban location. The use of these relatively inexpensive systems is potentially cardioprotective against PM2.5 exposures and warrants further research. ClinicalTrials.gov identifier: NCT03334565.


Atomistic simulation of hydrophobin HFBII conformation in aqueous and fluorous media and at the water/vacuum interface.
Hydrophobins are proteins of interest for numerous applications thanks to their unique conformational and surface properties and their ability to self-assemble at interfaces. Here we report fully atomistic molecular mechanics and molecular dynamics results together with circular dichroism experimental data, aimed to study the conformational properties of the hydrophobin HFBII in a fluorinated solvent in comparison with a water solution and/or at an aqueous/vacuum interface. Both the atomistic simulations and the circular dichroism data show the remarkable structural stability of HFBII at all scales in all these environments, with no significant structural change, although a small cavity is formed in the fluorinated solvent. The combination of theoretical calculations and circular dichroism data can describe in detail the protein conformation and flexibility in different solvents and/or at an interface, and constitutes a first step towards the study of their self-assembly.


General Versus Local Anesthesia With Conscious Sedation in Transcatheter Aortic Valve Implantation: The Randomized SOLVE-TAVI Trial.
In clinical practice, local anesthesia with conscious sedation (CS) is performed in roughly 50% of patients undergoing transcatheter aortic valve replacement. However, no randomized data assessing the safety and efficacy of CS versus general anesthesia (GA) are available. The SOLVE-TAVI (Comparison of Second-Generation Self-Expandable Versus Balloon-Expandable Valves and General Versus Local Anesthesia in Transcatheter Aortic Valve Implantation) trial is a multicenter, open-label, 2×2 factorial, randomized trial of 447 patients with aortic stenosis undergoing transfemoral transcatheter aortic valve replacement comparing CS versus GA. The primary efficacy end point was powered for equivalence (equivalence margin 10% with significance level 0.05) and consisted of the composite of all-cause mortality, stroke, myocardial infarction, infection requiring antibiotic treatment, and acute kidney injury at 30 days. The primary composite end point occurred in 27.2% of CS and 26.4% of GA patients (rate difference, 0.8 [90% CI, -6.2 to 7.8]; Pequivalence=0.015). Event rates for the individual components were as follows: all-cause mortality, 3.2% versus 2.3% (rate difference, 1.0 [90% CI, -2.9 to 4.8]; Pequivalence<0.001); stroke, 2.4% versus 2.8% (rate difference, -0.4 [90% CI, -3.8 to 3.8]; Pequivalence<0.001); myocardial infarction, 0.5% versus 0.0% (rate difference, 0.5 [90% CI, -3.0 to 3.9]; Pequivalence<0.001), infection requiring antibiotics 21.1% versus 22.0% (rate difference, -0.9 [90% CI, -7.5 to 5.7]; Pequivalence=0.011); acute kidney injury, 9.0% versus 9.2% (rate difference, -0.2 [90% CI, -5.2 to 4.8]; Pequivalence=0.0005). There was a lower need for inotropes or vasopressors with CS (62.8%) versus GA (97.3%; rate difference, -34.4 [90% CI, -41.0 to -27.8]). Among patients with aortic stenosis undergoing transfemoral transcatheter aortic valve replacement, use of CS compared with GA resulted in similar outcomes for the primary efficacy end point. These findings suggest that CS can be safely applied for transcatheter aortic valve replacement. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT02737150.


Yolk-sac carcinoma of extra-embryonic origin in the 129 Sv/Sl mouse.
Displacement of the visceral yolk-sac in fetectomized rats and hamsters does not only induce benign teratomas but may also lead to the development of malignant yolk-sac carcinoma. In the mouse, displacement of this extraembryonic membrane readily induces teratoma but until now yolk-sac carcinoma has never been observed in this species. In the present study we report the development of yolk-sac carcinoma in 129 Sv/Sl mice after a long latent period (8 months). The tumor is composed of visceral and parietal patterns in the primary neoplasm and in its metastases. It develops from the extra-embryonal part belonging to normal (+/+), heterozygous (Sl/+) or sterile (Sl/Sl) embryos, which proves that these yolk-sac carcinomas are not of germ-cell origin.


Differential Levels of Hippo Signaling in Selected Brain and Peripheral Tissues in Streptozotocin-Induced Cognitive Dysfunction in Mice.
Increasing studies have revealed that metabolic disorders, especially diabetes, are high risk factors for the development of Alzheimer's disease (AD) and other neurodegenerative diseases. It has been reported that patients with diabetes are prone to suffer from cognitive dysfunction (CD). Although abnormal glucose metabolism and deposition of amyloid β (Aβ) are proven to have a closely relationship with diabetes-induced CD, its exact mechanism is still undetermined. In this study, a total of 14 mice were intraperitoneally injected with streptozotocin for 5 consecutive days to mimic diabetic models, and then hierarchical cluster analysis was adopted to classify the diabetic mice into CD and Non-CD phenotypes by the results of Morris water maze test (MWMT). Furthermore, we detected Hippo signaling including mammalian sterile 20-like protein kinases1 (MST1), large tumor suppressors 1 (LATS1), Yes-associated protein (YAP) and phosphorylation of YAP (p-YAP) in brain and peripheral tissues. As compared with control mice, the levels of MST1, LATS1 and p-YAP/YAP ratio were increased in medial prefrontal cortex (mPFC), striatum and hippocampus of CD mice, while these proteins were decreased in gut tissue of CD mice. Additionally, there were significant positive correlations between escape latency and p-YAP/YAP ratio in mPFC, anterior cingulate cortex (ACC) and hippocampus, as well as the level of LATS1 in liver, kidney and gut tissues. In conclusion, alterations in Hippo signaling may contribute to CD induced by diabetes. Therefore, therapeutic interventions improving Hippo signaling might be beneficial to the treatment of diabetes-induced CD and other neurodegenerative diseases.


Mediators for the Effect of Compassion Cultivating Training: A Longitudinal Path Analysis in a Randomized Controlled Trial Among Caregivers of People With Mental Illness.
Background: There is a paucity of research on mediators of change, within compassion training programs. The aim was to investigate the mediators, of an 8-week compassion cultivation training (CCT) program, on the effect of psychological distress on caregivers of people with a mental illness. Method: Longitudinal path models in a randomized controlled trial (RCT). One hundred ninety-two participants were assessed for eligibility, and 161 participants were included into the trial and randomized. The main outcome was psychological distress measured by the Depression, Anxiety and Stress scale at 6 months. Mediators included self-compassion (SC), mindfulness (FM), emotion regulation (ER), emotion suppression (ES), and cognitive reappraisal (CR). Baseline, post, and 3- and 6-month follow-up measurements were collected. Results: The mediated effects for CCT are as follows: depression at 6 months: SC: -1.81 (95% CI: -3.31 to -0.31); FM: -1.98 (95% CI: -3.65 to -0.33); ER: -0.14 (95% CI: -1.31 to 1.02); anxiety at 6 months: SC: -0.71 (95% CI: -1.82 to 0.40); FM: -1.24 (95% CI: -2.39 to -0.09); ER: 0.18 (95% CI: -1.04 to 1.40); stress at 6 months: SC: -1.44 (95% CI: -2.84 to -0.05); FM: -2.17 (95% CI: -3.63 to -0.71); ER: -0.27 (95% CI: -1.51 to 0.98). Conclusion: Mindfulness and self-compassion are important components in reducing psychological distress experienced by informal caregivers of people with a mental illness. Results contribute to the knowledge about the underlying mechanisms of CCT.


Practical and low cost whole-organism motility assay: A step-by-step protocol.
Here, we provide a step-by-step protocol for a practical and low cost whole-organism assay for the screening of chemical compounds for activity against parasitic worms. This assay has considerable advantages over conventional methods, mainly in relation to ease of use, throughput, time and cost. It is readily suited to the screening of hundreds to thousands of compounds for subsequent hit-to-lead optimisation, and should be applicable to many different parasites and other organisms commensurate with the size of wells in the microtiter plates used for phenotypic screening.


Large fragment Sanger sequencing identifies the newly encountered variant that caused null alleles in parentage testing.
Short tandem repeat (STR) is regarded as a crucial tool for personal identification as well as parentage testing. Thus, genotyping errors of STRs could have negative effects on the reliability of forensic identification. A null allele at the combined DNA index system (CODIS) core loci D2S1338 was found in a father-daughter pair with the AGCU Expressmarker 22 kit which was a commonly used commercial kit during our daily laboratory work. This null allele caused the father and daughter to not conform to the laws of inheritance, thus potentially generating erroneous conclusions that excluded parentage. To figure out the reason for this phenomenon, re-amplification with new primers and then large fragment Sanger sequencing was conducted. We found a G to G/T variation at the position which is fifty-nine bases away from the 3' end of the core repeat in both samples. This probably could be considered a novel variant at the primer binding region which had not been reported that resulted in the emergence of the null allele. We also found that there was more than one single-nucleotide polymorphism (SNP) with minor allele frequency (MAF) greater than 0.1 in the upstream and downstream sequences of D2S1338. When designing primers for amplification of D2S1338, the possible adverse results of these SNPs should be taken into account and avoided.


Environmental biogeochemical behaviors of rare earth elements in soil-plant systems.
With the continual increase in the utilization of rare earth elements (REEs) for industrial and agricultural purposes in China, the research into the environmental biogeochemical behavior of REEs has become a pressing issue. The REEs' content in soil and various parts of wheat under different conditions in soil-plant systems were measured by INAA and ICP-MS. The results showed four aspects. (1) The mean value of total REEs in soil of China was 176.8 mg kg(-1). The mean ratio of SigmaLREE/SigmaHREE in soils was 8.0 and cerium accounts for 42% of the total REEs. The content of REEs in wheat seed ranged between 10(-11) and 10(-8) g g(-1), 3-4 orders of magnitude lower than that in soil. (2) The REEs contents in ryegrass, especially in roots, were significantly related to that of soil. The bioavailability of REEs in soil mainly depended on the exchangeable fraction of REEs, which was strongly affected by the physico-chemical properties of the soil. (3) Long-term foliage-dressing with Changle microfertilizer of REEs did not affect the contents and distribution patterns of REEs in soil. At the maturing stage of spring wheat, the REEs content was in the order of root > leaf >stem and crust. Compared with the control, foliage-dressing has a higher accumulation of REEs in root and leaf. However, no significant difference was found in stem and crust between the two treatments. (4) There was no significant accumulation with the soil-dressing method. When comparing controls in both foliage- and soil-dressing methods, no distinct residue of REEs in grains was found.


Prevalence of pharmaceuticals and personal care products, microplastics and co-infecting microbes in the post-COVID-19 era and its implications on antimicrobial resistance and potential endocrine disruptive effects.
The COVID-19 (coronavirus disease 2019) pandemic's steady condition coupled with predominance of emerging contaminants in the environment and its synergistic implications in recent times has stoked interest in combating medical emergencies in this dynamic environment. In this context, high concentrations of pharmaceutical and personal care products (PPCPs), microplastics (MPs), antimicrobial resistance (AMR), and soaring coinfecting microbes, tied with potential endocrine disruptive (ED) are critical environmental concerns that requires a detailed documentation and analysis. During the pandemic, the identification, enumeration, and assessment of potential hazards of PPCPs and MPs and (used as anti-COVID-19 agents/applications) in aquatic habitats have been attempted globally. Albeit receding threats in the magnitude of COVID-19 infections, both these pollutants have still posed serious consequences to aquatic ecosystems and the very health and hygiene of the population in the vicinity. The surge in the contaminants post-COVID also renders them to be potent vectors to harbor and amplify AMR. Pertinently, the present work attempts to critically review such instances to understand the underlying mechanism, interactions swaying the current health of our environment during this post-COVID-19 era. During this juncture, although prevention of diseases, patient care, and self-hygiene have taken precedence, nevertheless antimicrobial stewardship (AMS) efforts have been overlooked. Unnecessary usage of PPCPs and plastics during the pandemic has resulted in increased emerging contaminants (i.e., active pharmaceutical ingredients and MPs) in various environmental matrices. It was also noticed that among COVID-19 patients, while the bacterial co-infection prevalence was 0.2-51%, the fungi, viral, protozoan and helminth were 0.3-49, 1-22, 2-15, 0.4-15% respectively, rendering them resistant to residual PPCPs. There are inevitable chances of ED effects from PPCPs and MPs applied previously, that could pose far-reaching health concerns. Furthermore, clinical and other experimental evidence for many newer compounds is very scarce and demands further research. Pro-active measures targeting effective waste management, evolved environmental policies aiding strict regulatory measures, and scientific research would be crucial in minimizing the impact and creating better preparedness towards such events among the masses fostering sustainability.


Effects of carbamazepine on spinal cord ischemia.
Prophylactic treatment with carbamazepine has been shown to reduce the cerebral damage and neurologic deficit in ischemic conditions. A randomized controlled study based on a rabbit model was designed to study the effect of carbamazepine on a spinal cord ischemic reperfusion injury. Thirty New Zealand rabbits were randomly assigned to 1 of the 2 groups (n = 15 per group): group I (control group) and group II (carbamazepine group). Spinal cord ischemia was induced by infrarenal aortic crossclamp for 25 minutes in both groups. Functional evaluation with the Tarlov score during a 2-day observation period and histopathologic assessment of the lumbar spinal cord were performed. Changes in spinal cord morphology were observed with hematoxylin-eosin staining and electron microscopy. Gray matter damage was assessed on the basis of the number of normal neurons in the ventral horn. Diffuse destruction of gray matter with moderate to severe vacuolization and essentially no normal ganglion cells was observed in the spinal cord of rabbits in the control group, whereas specimens of rabbits assigned to the carbamazepine group showed ganglion cells with normal nuclei and cytoplasm (P < .0001). Neurologic impairment was significantly attenuated in the carbamazepine group compared with the Tarlov scores of the control group (P < .0001 at day 2). Carbamazepine may protect the spinal cord from ischemic reperfusion injury that is associated with ameliorated neurologic and histopathologic results.


Role of runt-related transcription factor 3 (RUNX3) in transcription regulation of natural cytotoxicity receptor 1 (NCR1/NKp46), an activating natural killer (NK) cell receptor.
Natural cytotoxicity receptor 1 (NCR1), also known as NKp46, is a natural killer (NK) lymphocyte-activating receptor. It is involved in major aspects of NK immune function and shows a high degree of lineage specificity in blood and bone marrow. The nature of its NK-restricted expression is not well understood. In this study, we confirm that human NCR1 NK-specific expression is achieved at the mRNA level. We found two key cis-regulatory elements in the immediate vicinity upstream of the gene. One element acts as an essential promoter, whereas the other acts as a tissue-dependent enhancer/repressor. This latter regulatory element contains a runt related-transcription factor (RUNX) recognition motif that preferentially binds RUNX3. Interfering with RUNX proteins using a dominant negative form results in decreased Ncr1 expression. RUNX3 overexpression had the opposite effect. These findings shed light on the role of RUNX3 in the control of an important NK-activating receptor.


Purin-6-yl 6-deoxy-1-thio-beta-D-glucopyranoside. Enzymology, chemistry, and cytotoxicity.
Purin-6-yl 6-deoxy-1-thio-beta-D-glucopyranoside (4) is a substrate for almond beta-glucosidase and a weak competitive inhibitor of bovine liver beta-D-glucuronidase (Ki approximately 20mM). Both 4 and purine-protonated 4 undergo hydrolysis catalyzed by dilute acid in the pH range 0.17-2.59. These results are compared with those previously obtained with ammonium (purin-6-yl 1-thio-beta-D-glucopyranosid)uronate, (purin-6-yl 1-thio-beta-D-glucopyranosid)uronamide, purin-6-yl 1-thio-beta-D-glucopyranoside, and purin-6-yl 2-deoxy-1-thio-beta-D-glucopyranoside, and it is concluded that the data support an involvement of substituents at C-5 in producing productive Michaelis-complex conformers. The 6-deoxyglucoside is more active than the D-glucosiduronic acid in an L1210 mouse screen.


Nonestrogen Therapies for Treatment of Genitourinary Syndrome of Menopause: A Systematic Review.
To systematically review the literature and provide clinical practice guidelines regarding various nonestrogen therapies for treatment of genitourinary syndrome of menopause (GSM). MEDLINE, EMBASE, ClinicalTrials.gov , and Cochrane databases were searched from inception to July 2021. We included comparative and noncomparative studies. Interventions and comparators were limited to seven products that are commercially available and currently in use (vaginal dehydroepiandrosterone [DHEA], ospemifene, laser or energy-based therapies, polycarbophil-based vaginal moisturizer, Tibolone, vaginal hyaluronic acid, testosterone). Topical estrogen, placebo, other nonestrogen products, as well as no treatment were considered as comparators. We double-screened 9,131 abstracts and identified 136 studies that met our criteria. Studies were assessed for quality and strength of evidence by the systematic review group. Information regarding the participants, details on the intervention and comparator and outcomes were extracted from the eligible studies. Alternative therapies were similar or superior to estrogen or placebo with minimal increase in adverse events. Dose response was noted with vaginal DHEA and testosterone. Vaginal DHEA, ospemifene, erbium and fractional carbon dioxide (CO 2 ) laser, polycarbophil-based vaginal moisturizer, tibolone, hyaluronic acid, and testosterone all improved subjective and objective signs of atrophy. Vaginal DHEA, ospemifene, tibolone, fractional CO 2 laser, polycarbophil-based vaginal moisturizer, and testosterone improved sexual function. Most nonestrogen therapies are effective treatments for the various symptoms of GSM. There are insufficient data to compare nonestrogen options to each other.


Temperature effect on growth rates of Eriophorum vaginatum roots.
Root growth rates of the sedge Eriophorum vaginatum L. were studied under controlled environmental conditions. The air temperature was maintained constant at 15°C while the root temperatures varied in 5°C intervals between 2° and 37° C (12° C excluded). Root growth rates of 1.2 mm d-1 at 2°, 20.4 mm d-1 at 32° C and 10.1 mm d-1 at 37° C were recorded. A Q10 of 3.2 was calculated for the temperature range from 7° to 27° C. Root growth rates at temperatures above 17° C declined after one week of growth. The degree of decline was proportional to the applied root temperature. Depletion of available nonstructural carbohydrate was the probable cause for this decline.


Extraction process optimization of Murraya koenigii leaf extracts and antioxidant properties.
The study was intended to optimise the process variables such as extraction time and solvent concentration to maximize the yield of Murraya koenigii leaf extract and total phenolic content using response surface methodology. The experimental design was conducted for independent factor such as acetone, ethanol, methanol (20-80%) and time (20-100 min). The optimal conditions as the quadratic model were retained through central composite design. All the variables showed significant influence on extract yield and total phenolic content of M. koenigii leaf extract. The optimized conditions of extract were attained as 50% of ethanol, 60% acetone, 80% methanol and further analysed for their DPPH scavenging activity, total phenolic content, flavonoid content, and ferric reducing activity. Extract obtained with 50% ethanol showed highest DPPH scavenging activity and total phenolic content while 60% acetonic extract exhibited highest ferric reducing activity and flavonoid content.


Three-dimensional models of the GDP and GTP forms of the guanine nucleotide domain of Escherichia coli elongation factor Tu.
Three-dimensional models of the GDP and GTP forms of the guanine nucleotide domain of Escherichia coli elongation factor Tu have been derived from the atomic coordinates of the trypsin-modified form of EF-Tu-GDP and by comparison with the ras p21 structures. The significance of the differences in the guanine nucleotide binding sites of EF-Tu and ras p21 are discussed. Crystallization of the EF-Tu-GMPPNP complex is reported.


Factors determining COVID-19 pneumonia severity in a country with routine BCG vaccination.
The impact of countries' bacillus Calmette-Guérin (BCG) vaccination policies on the course of coronavirus disease (COVID-19) outbreak is a curiosity. In this study, the relationship between BCG vaccination status and severity of COVID-19 pneumonia and the factors affecting disease severity were investigated. A retrospective cross-sectional study was conducted between March and June 2020 in patients diagnosed with COVID-19 pneumonia, confirmed by severe acute respiratory syndrome coronavirus-2 polymerase chain reaction positivity in a nasopharyngeal sample and pulmonary infiltrates in computed chest tomography, in a state hospital in Istanbul, Turkey. Socio-demographic features, body mass index, smoking status, concomitant diseases, income rates and BCG vaccination status of subjects were analyzed. The study population comprised 123 adults with COVID-19 pneumonia [mean age = 49·7 years, standard deviation = 13·3 years; 82 (66·7%) male]. While the rate of cases vaccinated with BCG is lower (68·5 versus 88·2%, P = 0·026), mean age (54·0 ± 11·5 years versus 38·3 ± 10·7 years; P < 0·001), diabetes (32·6 versus 5·9%, P = 0·002) and low income (84·3 versus 52·9%, P < 0·001) are higher in patients with severe disease compared to those with mild disease. According to multivariate analysis increasing age [odds ratio (OR) = 1·119; 95% confidence interval (CI) = 1·062-1·178, P < 0·001] and low income (OR = 3·209; 95% CI = 1·008-10·222, P = 0·049) are associated with severe disease in COVID-19 pneumonia. This study reveals that BCG vaccination is not associated with disease severity in COVID-19 pneumonia. Age and low income are the main determinants of severe COVID-19 pneumonia.


Effects of bile-acid-binding resin (colestimide) on blood glucose and visceral fat in Japanese patients with type 2 diabetes mellitus and hypercholesterolemia: an open-label, randomized, case-control, crossover study.
The objective was to examine the effects of colestimide on blood glucose, visceral fat, adipocytokines, and bile acid conjugate fractions in Japanese patients. This study was an open-label, randomized, case-control, crossover study of colestimide 3 g/day in 40 Japanese patients with type 2 diabetes mellitus (T2D) and hypercholesterolemia. Patients were assigned to the colestimide group in which pravastatin and colestimide were administered orally and to the statin group in which pravastatin alone was administered orally. The principal outcome measures were serum lipid levels, fasting plasma glucose level in the early morning, hemoglobin A1c (HbA(1c)), visceral fat area (VFA), and serum 1,5-anhydroglucitol (1,5-AG) level. Serum low-density lipoprotein cholesterol levels significantly decreased from 113±38 mg/dl at baseline to 90±20 mg/dl (P=.009) at week 12 of colestimide administration. HbA(1c) significantly decreased from 7.4%±0.9% at baseline to 6.9%±0.9% (P=.001) at week 12 of colestimide administration. Serum 1,5-AG levels increased from 9.4±10.1 μg/ml to 12.4±9.5 μg/ml (P=.05) at week 12 of colestimide administration. The statin group showed no significant changes in lipids and 1,5-AG. However, ΔVFA was inversely correlated with Δcholic acid, and multivariate analysis revealed that ΔVFA was a significant explanatory variable. Colestimide holds promise not only for the treatment of hypercholesterolemia but also for the possible improvement of T2D and visceral fat obesity.