Arterial enlargement, tortuosity, and intimal thickening in response to sequential exposure to high and low wall shear stress.
We investigated the effects of sequential and prolonged exposure to high and low wall shear stress on arterial remodeling using a rabbit arteriovenous fistula (AVF) model. Blood flow was increased by approximately 17-fold to 20-fold when the AVF was open, and returned to normal when the AVF was occluded. Repeated opening and closing of the AVF resulted in sequential exposure of the artery to high and low wall shear stress. High flow and high wall shear stress induced arterial dilatation, elongation, and tortuosity, without intimal thickening. The common carotid artery was elongated 37% after 4 weeks of high flow, and was shortened 10% after 6 weeks of normal flow. Subsequent cycles of high flow induced less elongation, with less shortening after return to normal flow. Enlargement of the distal segment was more dramatic than in the proximal segment, despite exposure to the same volume of flow and the same initial high wall shear stress after creation of the AVF. The distal carotid segment enlarged more than did the proximal segment during each exposure to high flow. In segments of carotid artery exposed to low wall shear stress (<5 dynes/cm(2)) intimal thickening developed. These changes were maximal in the distal carotid segment, just before the AVF. Each cycle of low wall shear stress induced intimal thickening accompanied by medial hyperplasia. Intimal thickening was inhibited during periods of high flow when wall shear stress was high. Three cycles of flow alteration induced three layers of intimal thickening in the distal arterial segment, two layers of intimal thickening in the middle segment, and one layer of intimal thickening in the proximal segment. Long-term exposure to low wall shear stress induced severe intimal thickening and medial hyperplasia in different segments. Thus the response of the carotid artery afferent to an AVF varies along the length of the artery, with maximum enlargement, elongation, and tortuosity in the distal segment, just proximal to the AVF. Similarly, intimal thickening in response to low wall shear stress is maximal in the distal carotid artery. It appears that intimal thickening is related to local levels of low wall shear stress, and occurs when wall shear stress chronically falls to less than 5 dynes/cm(2).


Effect of maternal protein deficiency on intrauterine development of small intestine with special reference to alkaline phosphatase.
Rat intestinal alkaline phosphatase activity was found to be greater in the fetus near term than in the progeny at birth. A set back in the orderly development of the enzyme was observed in the duodenum of pups born to protein-deficient mothers. Electrophoretic studies on polyacrylamide gel showed homogeneity of the enzyme in fetus and newborn pups. Severe intrauterine growth retardation was associated with reduced size of the enterocytes and their nuclei.


Bistable Glass-pattern motion reveals two different processes.
Examining the two motion processes is an elusive task due to the difficulty of finding a proper stimulus paradigm. A rotational Glass pattern created with a random-dot array by superimposing its rotated version on the top of it can provide such a paradigm. If we displace only its rotated part in the vertical or horizontal direction, a bistable motion occurs; local dot motion in the same direction and Glass-pattern motion in the orthogonal direction. From two experiments, we found local dot motion is predominant in short spatiotemporal range and global pattern motion in long spatiotemporal range. Since the stimulus allows us to maintain all of its properties identical except for the changes in spatiotemporal parameters, this result shows more robustly that the energy-based first-order motion favors short spatiotemporal ranges while the pattern-based second-order motion favors long spatiotemporal ranges.


Stent strut coverage of titanium-nitride-oxide coated stent compared to paclitaxel-eluting stent in acute myocardial infarction: TITAX-OCT study.
Drug-eluting stents (DES) have reduced the rate of restenosis but recent studies have raised concern over the risk of late stent thrombosis (LST). Incomplete stent endothelialization and delayed vascular healing have been associated with LST. The titanium-nitride-oxide coated bio-active stent (BAS) has shown promising results in patients with acute coronary syndromes, but there is little long-term optical coherence tomography (OCT) data comparing BAS with DES. The TITAX-AMI trial is a prospective, randomized, multicenter trial comparing BAS to paclitaxel-eluting stent (PES) in 425 patients with acute myocardial infarction. A total of 18 patients (9 per group) with no major cardiac events during follow-up, were enrolled in this substudy >36 months (mean 47 months) after stent implantation. Quantitative coronary angiography was performed and stent strut endothelialization and vascular healing were assessed with OCT. The binary stent strut coverage was significantly higher in the BAS group compared with the PES group (99.6 vs. 89.2%, p < 0.001) and there were less malapposed struts in the BAS group (0.2 vs. 13.8%, respectively, p < 0.001). The neointimal hyperplasia (NIH) thickness (266 ± 166 vs. 126 μm ± 126 μm, p < 0.001) and percentage of NIH area (26.2 vs. 7.6%, p < 0.001) were greater in the BAS group than in the PES group. Late incomplete endothelialization was not uncommon after PES implantation. Stents in the BAS group were completely endothelialized. This difference may contribute to the more common LST after PES implantation in the TITAX-AMI trial.


Anti-mGluR1 encephalitis: Case illustration and systematic review.
The literature for immune-mediated neurological disorders is evolving like no other field of neurological illnesses. Many new antibodies or disorders have been described in the last decade. The cerebellum is a brain structure susceptible to these immune-mediated pathologies, and anti-metabotropic glutamate receptor 1 (mGluR1) antibody has a predilection to the cerebellar tissue. Anti-mGluR1 encephalitis is a rare autoimmune disease affecting the central and peripheral nervous systems, triggering an acute or subacute cerebellar syndrome with varying degrees of severity. Anti-mGluR1 encephalitis is a rare autoimmune disease affecting the central nervous system. We aimed to systematically review reported cases of anti-mGluR1 encephalitis and summarize their clinical presentation, management, outcomes, and case reports. A search of the PubMed and Google Scholar databases was conducted and included all cases of anti-mGluR1 encephalitis published in English before October 1, 2022. A comprehensive systematic review was conducted using "metabotropic glutamate receptor type 1," "mGluR1," autoantibodies," "autoantibodies," "autoimmunity," and "antibody" as keywords. The risk of bias assessment of the evidence was performed using appropriate tools. The qualitative variables were presented as frequency and percentage. Including our case, 36 cases of anti-mGluR1 encephalitis (19 males, median age 52.5 years, 11.1% pediatric cases) have been reported. The most common clinical manifestations are ataxia, dysarthria, and nystagmus. Initial imaging was normal in 44.4% of patients; however, 75% of patients showed abnormality later in the disease course. The first-line therapy options include glucocorticoids, intravenous immunoglobulin, and plasma exchange. Rituximab is the most commonly used second-line treatment. Complete remission was achieved in only 22.2% of patients, and 61.8% were disabled by the end of their course. Anti-mGluR1 encephalitis manifests as symptoms of cerebellar pathology. Although the natural history has not been completely elucidated, early diagnosis with prompt initiation of immunotherapy could be imperative. Any patient suspected to have autoimmune cerebellitis should be tested for the presence of anti-mGluR1 antibody in the serum and cerebrospinal fluid. Escalation to an aggressive therapy approach should be applied in cases that do not respond to first-line therapies, and extended follow-up durations are required in all cases.


Evaluation of folate conjugated pegylated thermosensitive magnetic nanocomposites for tumor imaging and therapy.
Superparamagnetic iron oxide nanoparticles (SPIONs) have been receiving great attention lately in biomedical applications, such as in magnetic resonance imaging and drug delivery. However, their systemic administration still remains a challenge due to their hydrophobic nature with instances of aggregation leading to fast reticuloendothelial system (RES) uptake. In this study, magnetic nanocomposites with thermosensitive polymer have been investigated. Random polymers of N-isopropylacrylamide (NIPAAM), acrylic acid (AA) and PEGMA have been coated on SPIONs followed by conjugation with folic acid. Particles of ∼200 nm and low polydispersity 0.1-0.2 having a critical temperature (T(c)) of 44 °C were formed. Thermogravimetric and powder X-ray diffraction studies showed that the nanocomposites were composed of 90% cubic face-centered magnetite. Nearly 76.5% doxorubicin was loaded onto the nanoparticles by diffusion method. Drug release was higher at the hyperthermia temperature (72.42 ± 5.25% in 48 h) proving the thermoresponsive nature of the polymer. Folate conjugated samples showed a magnetization value of 32 emu/g as well as high r1 and r2 relaxivities in magnetic resonance imaging. R2 weighted images of nanocomposites were darker than the control with 20 μg/mL as the darkest. At this concentration the magnetic composites showed nearly 95% viability in L929 fibroblast cells. These thermoresponsive nanosystems with pegylated surfaces and size of ∼200 nm are therefore highly suitable for in vivo imaging and hyperthermia based drug delivery.


The activity and properties of a hepatic acid cholesteryl ester hydrolase obtained from rats of different age groups.
The activity of lysosomal acid cholesteryl ester hydrolase (acid CEH, EC 188.8.131.52) in rat liver was determined at 3, 5, 7, 10 and 20 wk following birth. The levels of acid CEH activity showed a marked decrease as rats grew older, whereas those of other lysosomal marker enzymes, such as acid phosphatase, beta-glucuronidase and cathepsin B and D, showed only a slight decrease. On the other hand, acid CEH activity was detected in all subcellular fractions obtained from rat liver, but the enzyme activity in these fractions did not show the age-related decrease observed in the lysosomal fraction. The results presented here suggest that the marked alteration of lysosomal acid CEH activity that accompanies aging may be related to its possible involvement in the regulation of cholesterol concentration in rat liver.


Diaryl naphthyl methanes a novel class of anti-implantation agents.
Diaryl naphthyl methanes and the corresponding 1, 2, 3, 4- and 5, 6, 7, 8-tetrahydro naphthyl methane derivatives have been synthesized as novel estrogen receptor binding ligands. The secondary and tertiary amino alkoxy derivatives of diaryl naphthyl and tetrahydro naphthyl methane interact with the estrogen receptor to elicit promising estrogenic, antiestrogenic and implantation inhibition activities in rats. The most active compounds in this series are 7, 9 and 20, cent percent active in preventing implantation in rats at 2.5 mgkg(-1) dose.


Hepatic mesenchymal hamartoma causing heart failure in the neonate.
This is the first report of an hepatic mesenchymal hamartoma causing congestive heart failure in an infant. These benign tumors usually present as a bulky abdominal mass but rarely can present with congestive failure when the angiomatous component is prominent.


A comparison between 2 absorbable hemostatic agents: gelatin sponge (Spongostan) and oxidized regenerated cellulose (Surgicel).
Oxidized, regenerated cellulose (Surgicel) and gelatin sponge (Spongostan) were packed in the sockets of upper third molars after surgical removal in 10 and 11 patients, respectively. The other side in each patient functioned as control. The study revealed that more pain was apparent on the sides, where the materials were packed, especially in the gelatin sponge group, although more patients preferred this material to oxidized, regenerated cellulose. The 2 materials did not differ with regard to swelling or bleeding. No complications were encountered. Use of these materials in post-extraction sockets did not seem to impair closure of the wound as estimated by epithelial cover of the sockets, although healing was slightly delayed in the gelatin group.


Opportunistic Invasive Infection by Group A Streptococcus During Anti-Interleukin-6 Immunotherapy.
Invasive group A Streptococcus (GAS) in immunocompetent individuals is largely linked to hypervirulent strains. Congenital immunodeficiencies and those acquired from chronic disease or immunosuppressant drugs also increase risk of severe illness. We recovered GAS from the blood of a patient receiving a biologic inhibitor of interleukin 6 (IL-6). Growth of this serotype M4 isolate in human blood or a murine bacteremia model was promoted by interleukin 1 or IL-6 inhibition. Hyperinvasive M1T1 GAS was unaffected by IL-6 in both models. These findings based on a natural experiment introduce IL-6 signaling deficiencies as a risk factor for invasive GAS.


Organ Donation after Circulatory Determination of Death in India: A Joint Position Paper.
Organ donation following circulatory determination of death (DCDD) has contributed significantly to the donor pool in several countries. In India, majority of deceased donations happen following brain death (BD). While existing legislation allows for DCDD, there have been only few reports of kidney transplantation following DCDD from India. This document, prepared by a multidisciplinary group of experts, reviews international best practices in DCDD and outlines the path for DCDD in India. Ethical, medical, legal, economic, procedural, and logistic challenges unique to India have been addressed. The practice of withdrawal of life-sustaining treatment (WLST) in India, laid down by the Supreme Court of India, is time-consuming, possible only in patients in a permanent vegetative state, and too cumbersome for day-to-day practice. In patients where continued medical care is futile, the procedure for WLST is described. In controlled DCDD (category-III), decision for WLST is independent of and delinked from the subsequent possibility of organ donation. Families that are inclined toward organ donation are explained the procedure including the timing and location of WLST, consent for antemortem measures, no-touch period, and the possibility of stand-down and return to the intensive care unit (ICU) without donation. In donation following neurologic determination of death (DNDD), if cardiac arrest occurs during the process of BD declaration, the protocol for DCDD category-IV has been described in detail. In DCDD category-V, organ donation may be possible following unsuccessful cardiopulmonary resuscitation of cardiac arrest in the ICU. An outline of organ-specific requisites for kidney, liver, heart, and lung transplantation following DCDD and techniques, such as normothermic regional perfusion (nRP) and ex vivo machine perfusion, has been provided. The outcomes of transplantation following DCDD are comparable to those following DBDD or living donor transplantation. Documents and checklists necessary for successful execution of DCDD in India are described. Seth AK, Mohanka R, Navin S, Gokhale AGK, Sharma A, Kumar A, et al. Organ Donation after Circulatory Determination of Death in India: A Joint Position Paper. Indian J Crit Care Med 2022;26(4):421-438.


Development of a time-stepping sediment budget model for assessing land use impacts in large river basins.
The use of river basin modelling to guide mitigation of non-point source pollution of wetlands, estuaries and coastal waters has become widespread. To assess and simulate the impacts of alternate land use or climate scenarios on river washload requires modelling techniques that represent sediment sources and transport at the time scales of system response. Building on the mean-annual SedNet model, we propose a new D-SedNet model which constructs daily budgets of fine sediment sources, transport and deposition for each link in a river network. Erosion rates (hillslope, gully and streambank erosion) and fine sediment sinks (floodplains and reservoirs) are disaggregated from mean annual rates based on daily rainfall and runoff. The model is evaluated in the Burdekin basin in tropical Australia, where policy targets have been set for reducing sediment and nutrient loads to the Great Barrier Reef (GBR) lagoon from grazing and cropping land. D-SedNet predicted annual loads with similar performance to that of a sediment rating curve calibrated to monitored suspended sediment concentrations. Relative to a 22-year reference load time series at the basin outlet derived from a dynamic general additive model based on monitoring data, D-SedNet had a median absolute error of 68% compared with 112% for the rating curve. RMS error was slightly higher for D-SedNet than for the rating curve due to large relative errors on small loads in several drought years. This accuracy is similar to existing agricultural system models used in arable or humid environments. Predicted river loads were sensitive to ground vegetation cover. We conclude that the river network sediment budget model provides some capacity for predicting load time-series independent of monitoring data in ungauged basins, and for evaluating the impact of land management on river sediment load time-series, which is challenging across large regions in data-poor environments.


Donor-Recipient Matching to Optimize the Utility of High Kidney Donor Profile Index Kidneys.
In December 2014, the Kidney Donor Profile Index (KDPI) was developed to give more precise information on donor kidney quality. Kidneys with KDPI scores ≥ 85 (K ≥ 85) have been reported to have inferior outcomes to kidneys with KDPI scores < 85. After the implementation of the new Kidney Allocation System, we developed a protocol to evaluate K ≥ 85 use. We analyzed the safety and efficacy of our institutional criteria and evaluated post-transplant outcomes. K ≥ 85 recipients were stratified based on their 1-year creatinine and estimated glomerular filtration rates to elucidate characteristics associated with serum creatinine < 1.7 mg/dL or estimated glomerular filtration rates ≤ 45 mL/min/1.73 m2. From December 2014 to December 2019, 304 deceased donor kidney transplants were performed at Hartford Hospital; 58 (19%) were K ≥ 85 with an average KDPI of 91%. There were 4 graft losses; 2 were death censored. Prolonged cold ischemia time and black recipient race were associated with inferior recipient graft function at 1 year. High KDPI kidney use requires a multifaceted evaluation that takes into account donor and recipient characteristics for an ideal match. We have identified several characteristics that may predict optimal post-transplant kidney function.


[A retrospective analysis of 8 cases with primary central nervous system lymphoma].
To make an inquiry into clinical features, treatment, and prognosis of primary central nervous system lymphoma (PCNSL). A retrospective analysis was carried out in a group of 8 patients confirmed diagnostic PCNSL by clinical findings and pathological examination. The common clinical manifestation of PCNSL consisted of increased intracranial pressure, seizures, focal neurologic deficits. Studies of cerebrospinal fluid revealed protein elevation in the most patients. Neuroradiological examinations with CT and MRI both showed solitary or multiple lesion located cerebral hemisphere, periventricular, basal ganglia, corpus callosum. Pathological examination showed B-cell malignant lymphoma. The patients with PCNSL usually did not show characteristic clinical manifestations or findings in imaging scanning. The brain biopsy pathological examination might be the only reliable method to confirm the diagnosis of the disease.


Gene Editing and Human Pluripotent Stem Cells: Tools for Advancing Diabetes Disease Modeling and Beta-Cell Development.
This review will focus on the multiple approaches to gene editing and address the potential use of genetically modified human pluripotent stem cell-derived beta cells (SC-β) as a tool to study human beta-cell development and model their function in diabetes. We will explore how new variations of CRISPR/Cas9 gene editing may accelerate our understanding of beta-cell developmental biology, elucidate novel mechanisms that establish and regulate beta-cell function, and assist in pioneering new therapeutic modalities for treating diabetes. Improvements in CRISPR/Cas9 target specificity and homology-directed recombination continue to advance its use in engineering stem cells to model and potentially treat disease. We will review how CRISPR/Cas9 gene editing is informing our understanding of beta-cell development and expanding the therapeutic possibilities for treating diabetes and other diseases. Here we focus on the emerging use of gene editing technology, specifically CRISPR/Cas9, as a means of manipulating human gene expression to gain novel insights into the roles of key factors in beta-cell development and function. Taken together, the combined use of SC-β cells and CRISPR/Cas9 gene editing will shed new light on human beta-cell development and function and accelerate our progress towards developing new therapies for patients with diabetes.


Tp-e interval and Tp-e/QTc ratio as novel surrogate markers for prediction of ventricular arrhythmic events in hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) as a common genetic heart disease characterized by ventricular hypertrophy and myocardial fibrosis is significantly associated with a higher risk of fatal ventricular arrhythmic events (VAEs). We aimed to assess the interval between the peak and the end of the electrocardiographic T wave (Tp-e) and Tp-e/corrected QT (QTc) ratio as candidate markers of ventricular arrhythmias in patients with HCM. In this single-center, prospective study, a total of 66 patients with HCM and 88 controls were enrolled. The patients were divided into two groups: those with VAEs (n=26) and those without VAEs (n=40). Tp-e interval and Tp-e/QTc ratio were measured using a 12-lead electrocardiogram. Tp-e interval was significantly longer and Tp-e/QTc ratio were significantly higher in HCM patients than in the controls. In correlation analysis, maximal left ventricular (LV) thickness also has a significant positive correlation with Tp-e interval (r=0.422, p<0.001) and Tp-e/QTc ratio (r=0.348, p<0.001). Finally, multivariable regression analysis showed that a history of syncope, Tp-e interval [OR (odds ratio): 1.060; 95% confidence interval (CI): 1.005-1.117); p=0.012], Tp-e/QTc ratio (OR: 1.148; 95% CI: 1.086-1.204); p=0.049], and maximal LV thickness were independent predictors of VAEs in patients with HCM. Our findings suggested that prolonged Tp-e interval and increased Tp-e/QTc ratio may be good surrogate markers for the prediction of VAEs in HCM.


Quantifying the impacts of climatic trend and fluctuation on crop yields in northern China.
Climate change plays a critical role in crop yield variations, which has attracted a great deal of concern worldwide. However, the mechanisms of how climatic trend and fluctuations affect crop yields are not well understood and need to be further investigated. Thus, using the GIS-based Environmental Policy Integrated Climate (EPIC) model, we simulated the yields of major crops (i.e., wheat, maize, and rice) and evaluated the impacts of climatic factors on crop yields in the Agro-Pastoral Transitional Zone (APTZ) of northern China between 1980 and 2010. The partial least squares regression model was used to assess the contribution rates of climatic factors (i.e., precipitation, photosynthetically active radiation (PAR), minimum temperature (T min), maximum temperature (T max)) to the variation of crop yields. The Breaks for Additive Season and Trend (BFAST) model was adopted to decompose the climate factors into trend and fluctuation components, and the relative contributions of climate trend and fluctuation were then evaluated. The results indicated that the contributions of climatic factors to yield variations of wheat, maize, and rice were 31.7, 37.7, and 23.1%, respectively. That is, climate change had larger impacts on maize than wheat and rice. More cultivated areas were significantly and positively correlated with precipitation than with other climatic factors due to the limited precipitation in the APTZ. Also, climatic trend component had positive impacts on crop yields in the whole region, whereas the climate fluctuation was associated mainly with the areas where the crop yields decreased. This study helps improve our understanding of the mechanisms of climate change impacts on crop yields, and provides useful scientific information for designing regional-scale strategies of adaptation to climate change.


Regulation of type I collagen synthesis by 1,25-dihydroxyvitamin D3 in human osteosarcoma cells.
Synthesis of type I and III collagens has been examined in MG-63 human osteosarcoma cells after treatment with the steroid hormone, 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). Analysis of total [3H]proline-labeled proteins and pepsin-derived collagens revealed that 1,25-(OH)2D3 selectively stimulated synthesis of alpha 1I and alpha 2I components of type I collagen after 6-12 h. Consistent with previous reports (Franceschi, R. T., Linson, C. J., Peter, T. C., and Romano, P. R. (1987) J. Biol. Chem. 262, 4165-4171), parallel increases in fibronectin synthesis were also observed. Hormonal effects were maximal (2- to 2.5-fold versus controls) after 24 h and persisted for at least 48 h. In contrast, synthesis of the alpha 1III component of type III collagen was not appreciably affected by hormone treatment. Of several vitamin D metabolites (1,25-(OH)2D3, 25-dihydroxyvitamin D3, and 24R,25-dihydroxyvitamin D3) tested for activity in stimulating type I collagen synthesis, 1,25-(OH)2D3 was found to be the most active. Analysis of collagen mRNA abundance by Northern blot hybridization indicated that both types I and III procollagen mRNAs were increased 4-fold after a 24-h exposure to 1,25-(OH)2D3. Pro alpha 1I mRNA remained elevated through the 48-h time point while pro alpha 2I and pro alpha 1III mRNAs returned to control values. These results indicate that the regulation of collagen synthesis by 1,25-(OH)2D3 is complex and may involve changes in translational efficiency as well as mRNA abundance. 1,25-(OH)2D3 also caused at least a 20-fold increase in levels of the bone-specific calcium-binding protein, osteocalcin. These results are consistent with the hypothesis that 1,25-(OH)2D3 is stimulating partial differentiation to the osteoblast phenotype in MG-63 cells.


Fractionation and identification of metalloproteins from a marine cyanobacterium.
Trace metals are essential for the growth of marine cyanobacteria, being required for key cellular processes such as photosynthesis and respiration. Despite this, the metalloproteomes of marine cyanobacteria are at present only poorly defined. In this study, we have probed the major cobalt, iron, manganese, and nickel-binding proteins in the marine cyanobacterium Synechococcus sp. WH8102 by using two different fractionation approaches combined with peptide mass fingerprinting. For the identification of intact metalloproteins, multidimensional native chromatography was used to fractionate the proteome, followed by inorganic mass spectrometry to identify metal-enriched fractions. This approach led to the detection of nickel superoxide dismutase together with its predicted cofactor. We also explored the utility of immobilized metal affinity chromatography (IMAC) to isolate subpopulations of proteins that display affinity for a particular metal ion. We conclude that low-resolution 2D liquid chromatography is a viable fractionation technique to correlate relatively low-abundance metal ions with their few cellular destinations (e.g. Ni), but challenges remain for more abundant metals with multiple destinations such as iron. IMAC has been shown as a useful pre-fractionation technique to screen for proteins with metal-binding capacity, and may become a particularly valuable tool for the identification of metal-trafficking proteins.


Preservation of posterior mandibular extraction site with allogeneic demineralized, freeze-dried bone matrix and calcium sulphate graft binder before eventual implant placement: a case series.
This case series reports short- and long-term healing, before and after placement of an implant, in posterior mandibular extraction sites grafted with demineralized, freeze-dried bone matrix (DFDBM) allograft mixed with calcium sulphate graft binder. Three patients who underwent surgical extraction of a posterior mandibular molar experienced partial loss of the buccal bone plate at the extraction site. Alveolar bone reconstruction with a DFDBM allograft mixed with calcium sulphate graft binder was performed immediately. The graft was covered with a biodegradable regenerative membrane. For each of the 3 patients, the implant and healing abutment were placed after 6, 9 and 12 months, respectively, followed by crown placement 3, 5 and 5 months later, respectively. The implants were periodically re-evaluated, both clinically and radiographically, between 10 and 39 months after final insertion of the crown. An implant stability device was used to evaluate the long-term biological and functional stability of the implants. Upon exposure and implant placement, the grafted alveolar ridge in all patients presented appropriately sized, dense and well-vascularized bone, wide enough to receive the planned wide-platform implant. The long-term interface stability quotient ranged from 87 to 90. Posterior mandibular extraction sites with compromised buccal alveolar bone may be effectively managed by immediate alveolar augmentation using a mixture of DFDBM allograft and calcium sulphate graft binder. This approach provides ideal alveolar form and consistency for eventual placement of the implant.


Development of a simple, rapid in vitro cellular assay for bovine tuberculosis based on the production of gamma interferon.
Whereas in vitro assays of antibody responses to infectious agents are commonly used for routine diagnostic purposes, diagnostic tests for cellular responsiveness are confined to in vivo intradermal tests. This paper describes a simple and rapid in vitro cellular assay for bovine tuberculosis. The assay system is based on the detection of gamma interferon (gamma IFN), which is released in response to specific antigen. This assay can be carried out with whole blood samples thus avoiding the time-consuming task of isolating lymphocytes. A simple bioassay has been developed to quantitate the amount of gamma IFN produced, but this will eventually be replaced with an enzyme-linked immunoassay using monoclonal antibodies specific for bovine gamma IFN. The application of this system to bovine tuberculosis and other infectious diseases may provide a convenient in vitro cellular assay for routine diagnostic purposes.


Involvement of a Hydrophobic Pocket and Helix 11 in Determining the Modes of Action of Prenylated Flavonoids and Isoflavonoids in the Human Estrogen Receptor.
Six prenylated (iso)flavonoids were purified from a licorice root extract and subjected to competition experiments with six commercially available (iso)flavonoids. The agonistic and antagonistic activities of these compounds towards both hERα (human estrogen receptor alpha) and hERβ were determined. Differences in the modes of action (agonist or antagonist) were observed for the various compounds tested. In general, each compound had the same mode of action towards both ERs. In silico modeling was performed in order to study the differences in estrogenicity observed between the compounds. It is suggested that prenyl chains fit into a hydrophobic pocket present in the hER, resulting in an increased agonistic activity. In addition, it was shown that an increase in length (≈1.7 Å) of pyran prenylated isoflavonoids resulted in an antagonistic mode of action. This might be caused by collision of the pyran ring with helix 11 in the ligand binding cavity of the hER.


Systematic Scoping Review of Studies Reporting Unexpected Donor-Derived Abnormalities from Recipients of Allogeneic Hematopoietic Cell Transplantation: A Proposed Framework for Donor Disclosure.
Allogeneic hematopoietic cell transplantation (HCT) is used increasingly to treat blood and immune-based disorders. Post-transplantation testing of HCT recipients can lead to unexpected molecular, cytogenetic, and other information in donor-derived cells, raising questions regarding the potential impact on donor health. This study was conducted to identify the breadth of donor-derived abnormalities identified by testing HCT recipients and to determine the extent to which disclosure and donor follow-up are described. A systematic search and scoping review were conducted following PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) extension for scoping review guidelines in OVID MEDLINE and Embase (1947 to May 24, 2021). We identified 38 studies (63 donor-recipient pairs) addressing nonleukemic abnormalities to complement existing literature describing donor cell leukemia and donor-derived myelodysplasia. Donors were unrelated adults (n = 20), related family members (n = 28), cord blood donors (n = 6), or not reported (n = 9). Acquired cytogenetic, molecular, and morphologic abnormalities were reported. Donor origin was confirmed by cytogenetic analysis via karyotyping, fluorescence in situ hybridization, single tandem repeat PCR, and other techniques. A disease in donor-derived cells was described in 35 recipients (56.5%). Despite the relevance for testing and disclosure to donors, only 22 cases (32%) mentioned donor follow-up, and in 5 cases the donor developed a disease associated with the identified abnormality. Unrelated donor disclosure was mentioned in 3 of 26 cases (12%), with the findings reported back to the registry. Incidental abnormalities identified in transplanted donor cells may contribute to the post-transplantation risk of illness in the recipient and may be relevant to donor health. A framework for donor disclosure is proposed that incorporates consideration of analytic validity of the testing, potential significance of the finding, and the extent to which the abnormality is actionable. Adoption of effective processes to safeguard both donor and recipient health outcomes related to this issue is needed.


Circular dichroism detection in high-performance liquid chromatography: evaluation of the anisotropy factor.
The application of a circular dichroism (c.d.) detection system in HPLC using a chiral stationary phase is presented. The simultaneous measurement of the absorbance and c.d. signal allows the evaluation of the anisotropy factor (g = delta epsilon/epsilon) and thus the determination of the enantiomeric excess (e.e.) of the eluates. When this detection system is used in preparative chiral chromatography the collection of the enantiomeric fractions can be readily optimized.


Localized orbital locator as a descriptor for quantification and digital presentation of lone pairs: benchmark calculations of 4-substituted pyridines.
The parameters of the (3,-3) critical point in the localized orbital locator topology near a heteroatom have been found to reflect the changes in the size, density and electron energy of the lone pair and correlate with the donor ability of the lone pair carrying heteroatom.


The necessity and the value of placebo.
The use of placebo in clinical trials has been repeatedly challenged as being unacceptable from an ethical point of view. The present paper responds to this criticism by taking up the issue in the light of the pertinent provisions of the Helsinki Declaration. Examples from different therapeutic areas are given that highlight the importance of placebo in situations in which its use is acceptable according to the Declaration. Particular emphasis is given to the question of active control trials, which, under conditions of low assay sensitivity, may become an ethically less acceptable approach than the use of a placebo control.


Effectiveness of oral cephalexin in antibiotic-course completion for methicillin-susceptible Staphylococcus aureus-induced bacteremic vertebral osteomyelitis.
Methicillin-susceptible Staphylococcus aureus (MSSA) is the most common causative microorganism of pyogenic vertebral osteomyelitis (PVO). Although oral antimicrobial therapy with first-generation cephalosporins can treat MSSA infection, data on PVO are scarce. This study evaluated the treatment efficacy of cephalexin as oral antibiotic therapy for MSSA-induced PVO. This retrospective study included adult patients treated with oral cephalexin as the completing treatment for PVO with MSSA bacteremia from 2012 to 2020. Treatment effectiveness of cephalexin was evaluated by comparing improvement (5-point scale; score ≥ 4/5 indicates treatment success) in symptoms and laboratory and imaging results between intravenous antimicrobial and oral cephalexin treatment. Among 15 participants (8 [53%] women; median [interquartile range, IQR], age 75 [67.5-80.5] years; Charlson Comorbidity Index 2 [0-4]), 10 (67%) had lumbar spine lesions, 12 (80%) had spinal abscesses, and 4 (27%) had remote abscesses; no patients had concomitant endocarditis. In 11 patients with normal renal function, cephalexin 1,500-2,000 mg/day was administered. Five patients (33%) underwent surgery. Median (IQR; range) duration (days) of intravenous antibiotics, cephalexin, and total treatment was 36 (32-61; 21-86), 29 (19-82; 8-251), and 86 (59-125; 37-337), respectively. Cephalexin had an 87% treatment success rate without recurrence during a median follow-up of 119 (IQR, 48.5-350) days. In patients with MSSA bacteremia and PVO, antibiotic treatment completion with cephalexin is a reasonable option, even in cases with spinal abscess, if at least 3 weeks of effective intravenous antimicrobial therapy is provided.


Fat pad-derived mesenchymal stem cells as a potential source for cell-based adipose tissue repair strategies.
Mesenchymal stem cells are able to undergo adipogenic differentiation and present a possible alternative cell source for regeneration and replacement of adipose tissue. The human infrapatellar fat pad is a promising source of mesenchymal stem cells with many source advantages over from bone marrow. It is important to determine whether a potential mesenchymal stem-cell exhibits tri-lineage differentiation potential and is able to maintain its proliferation potential and cell-surface characterization on expansion in tissue culture. We have previously shown that mesenchymal stem cells derived from the fat pad can undergo chondrogenic and osteogenic differentiation, and we characterized these cells at early passage. In the study described here, proliferation potential and characterization of fat pad-derived mesenchymal stem cells were assessed at higher passages, and cells were allowed to undergo adipogenic differentiation. Infrapatellar fat pad tissue was obtained from six patients undergoing total knee replacement. Cells isolated were expanded to passage 18 and proliferation rates were measured. Passage 10 and 18 cells were characterized for cell-surface epitopes using a range of markers. Passage 2 cells were allowed to undergo differentiation in adipogenic medium. The cells maintained their population doubling rates up to passage 18. Cells at passage 10 and passage 18 had cell-surface epitope expression similar to other mesenchymal stem cells previously described. By staining it was revealed that they highly expressed CD13, CD29, CD44, CD90 and CD105, and did not express CD34 or CD56, they were also negative for LNGFR and STRO1. 3G5 positive cells were noted in cells from both passages. These fat pad-derived cells had adipogenic differentiation when assessed using gene expression for peroxisome proliferator-activated receptor γ2 and lipoprotein lipase, and oil red O staining. These results indicate that the cells maintained their proliferation rate, and continued expressing mesenchymal stem-cell markers and pericyte marker 3G5 at late passages. These results also show that the cells were capable of adipogenic differentiation and thus could be a promising source for regeneration and replacement of adipose tissue in reconstructive surgery.


[Diagnosis and treatment of gastric carcinoid tumors].
To investigate the method for surgical diagnosis and treatment of gastric carcinoid tumors. Clinicopathologic features of 9 patients with gastric carcinoid tumors were analyzed retrospectively. The patients were confirmed by biopsy at endoscopy or surgery. Eight of the 9 patients underwent elective operation, 6 showed gastric carcinoid tumors before operation found by biopsy at endoscopy. Five tumors were malignant and 4 benign. Two patients showed malignant carcinoid syndrome, but 5-HT and 5-HIAA were normal. Benign or malignant gastric carcinoid tumor depends upon tumor size, local lymph node invasion and distant metastases. The prognostic factor is based on benign or malignant tumors. Operation may be the first choice for such patients.


HIV1-viral protein R (Vpr) mutations: associated phenotypes and relevance for clinical pathologies.
Over the last 30 years, research into HIV has advanced the knowledge of virus genetics and the development of efficient therapeutic strategies. HIV-1 viral protein R (Vpr) is a specialized and multifunctional protein that plays important roles at multiple stages of the HIV-1 viral life cycle. This protein interacts with a number of cellular and viral proteins and with multiple activities including nuclear transport of the pre-integration complex (PIC) to the nucleus, transcriptional activation, cell cycle arrest at G2/M transition phase and induction of cell death via apoptosis. Specifically, Vpr has been shown to control many host cell functions through a variety of biological processes and by interaction with several cellular pathways. The different functions of Vpr may enhance viral replication and impair the immune system in HIV-1 infected patients. Importantly, functional defects induced by mutations in the Vpr protein correlate with slow disease progression of HIV-infected patients. Vpr is also associated with other concomitant pathologies developed by these patients, which may lead it to be considered as a potential novel therapeutic target. This review will focus on HIV-1 Vpr, mainly on the importance of its structural mutations on the progression of HIV infection, associated phenotypes and relevance for clinical pathologies. Copyright © 2016 John Wiley & Sons, Ltd.


Large-scale, closed-loop interrogation of neural circuits underlying cognition.
Cognitive functions are increasingly understood to involve coordinated activity patterns between multiple brain regions, and their disruption by neuropsychiatric disorders is similarly complex. Closed-loop neurostimulation can directly modulate neural signals with temporal and spatial precision. How to leverage such an approach to effectively identify and target distributed neural networks implicated in mediating cognition remains unclear. We review current conceptual and technical advances in this area, proposing that devices that enable large-scale acquisition, integrated processing, and multiregion, arbitrary waveform stimulation will be critical for mechanistically driven manipulation of cognitive processes in physiological and pathological brain networks.


Nitric oxide levels in the aqueous humor in cataract patients.
To determine the role of nitric oxide (NO) in the clinical manifestations of cataract in different age and etiological groups. Multicenter study, Taipei, Taiwan, Republic of China. Aqueous humor samples were obtained from 114 cataract patients in different etiological and age groups (eg, juvenile, traumatic, and age-related cataract). Using chemiluminescence assay, NO was detected through the light generated by ozone and NO interaction. The amount of NO production was calculated and compared among groups. The results were correlated with patient age. The NO levels in aqueous humor were highest in the 12 patients with traumatic cataract and lowest in the 15 patients with juvenile cataract (47.59 micromol/L +/- 12.81 [SD] and 7.66 +/- 2.62 micromol/L, respectively; P <.001). Aqueous humor NO levels were correlated with age and were highest in patients older than 80 years (mean 38.78 +/- 6.29 micromol/L) and in those with mature cataract (mean 40.15 +/- 6.15 micromol/L) (P <.05). The NO levels in the aqueous humor increased with age and traumatic events. The results suggest that NO could be a risk factor in cataract formation.


Chloride intracellular channel 1 (CLIC1) is activated and functions as an oncogene in pancreatic cancer.
Chloride intracellular channel 1 (CLIC1), a newly discovered member of the chloride channel protein family, has been implicated in multiple human cancers. However, little is known with regard to its expression and biological functions in pancreatic cancer. In this study, we focused on the clinical significance and biological functions of CLIC1 in pancreatic cancer and found that this protein was overexpressed in pancreatic cancer tissues. Patients with CLIC1-positive tumours had worse overall survival than those with CLIC1-negative tumours. Furthermore, the treatment of pancreatic cancer cell lines with CLIC1-targeting siRNA oligonucleotides significantly reduced cell proliferation and diminished anchorage-independent growth on both soft agar and cell migration. These data indicate that CLIC1 acts as a putative oncogene in pancreatic cancer and may represent a novel diagnostic and therapeutic target for pancreatic cancer.


Assessments of salivary antioxidant activity using electron spin resonance spectroscopy.
In recent years, the function of saliva has been focused on evaluation of general status. The relationship between salivary antioxidant activity and periodontal disease progression is unclear. The aim of this study is to assess the relationship between periodontal disease and salivary antioxidant activity towards various reactive oxygen species (ROS) using electron spin resonance (ESR) technique. We demonstrated that whole saliva derived rats or human subjects scavenged ROS such as superoxide (O(2)(·-)) and hydroxyl radical (HO(·)) using ESR spectroscopy with spin trapping agent. In addition, we assessed the relationship between antioxidants activity towards ROS and periodontal index with superoxide dismutase (SOD) activity in human subject saliva. Antioxidant activity towards O(2)(·-) was increased by Porphyromonas gingivalis (P. gingivalis) infection in rat, although antioxidant activity towards HO(·) was not changed. In human, a strong correlation (r = 0.88, p < 0.01) recognized between salivary antioxidant activity towards O(2)(·-) and probing pocket depth (PPD). In addition, the intensity of salivary antioxidant activity depended on SOD activity level. SOD activity was also correlated with PPD. Rat salivary antioxidant activity towards O(2)(·-) was up-regulated by the inflammatory response caused by P. gingivalis infection. Similar response was recognized in human saliva with periodontal index. Additionally, a linear correlation between antioxidant activity towards O(2)(·-) and SOD activity was verified by ESR technique. Therefore, evaluation of the salivary antioxidant activity towards O(2)(·-) might be an effective parameter for the objective assessment of periodontal disease progression.


Movement of internal canalicular orifice in association with blinking: direct observation after dacryocystorhinostomy.
To examine movement of the internal canalicular orifice with blinking and lacrimal drainage under endonasal endoscopic observation after dacryocystorhinostomy. Observational anatomic study. Twenty internal canalicular orifices (right 9, left 11) from 15 patients (age range: 44-77 years) who underwent endoscopic endonasal dacryocystorhinostomy were prospectively examined. The patients sat on a chair with the chin projected slightly upward while digital images were taken. After examining the movement of the internal canalicular orifice with blinking, the patient's eye was stained with fluorescein dye, and diffusion from the orifice was examined with blinking. The internal canalicular orifice closed during eyelid opening, although always incompletely. In eyelid closing, the orifice opened, and was pulled laterally without folds. All 20 internal canalicular orifices formed a diverticulum. Three specimens illustrated the upper and lower canalicular ends emptying into the diverticulum. The closing of these canalicular ends was always incomplete. Movement of the internal canalicular orifice was slight during normal blinking, but forced blinking resulted in more movement. Movement of the lacrimal sac wall was slight. During the first few seconds without blinking, no fluorescein dye flowed from the orifice. After several blinks, fluorescein dye flowed out slowly from the orifice, and increased in volume with more blinking. The fluorescein dye traveled inferiorly by gravity. A forced blinking was related to more dye inflow. The internal canalicular orifice incompletely closed during eyelid opening, but this orifice largely opened during eyelid closing, with a slow gravitational inflow of lacrimal fluid.


Molecular insights into how SHBG dimerization exerts changes on ligand molecular recognition.
Sex hormone binding globulin (SHBG) is a homodimeric glycoprotein and is the major carrier protein for sex steroids in plasma, regulating sex hormone availability in most vertebrate groups. Although it was initially thought that human dimeric SHBG bound a single ligand at the homodimer interface, studies demonstrated that dimeric SHBG binds a ligand to each subunit with similar affinity. In fact, the findings from recent experimental studies suggest that ligand binding to the SHBG dimer involves a complex allosteric mechanism involving conformational changes that limit observations of the presence of allosteric regulation. Therefore, we combined structural data with molecular dynamics simulations using Molecular Mechanics Generalized-Born Surface Area (MMGBSA) to dissect the structural and energetic basis for molecular recognition between five ligands whose affinities and binding positions on SHBG are known, i.e., 3β,17α-diol; 3β,17β-diol; DHT; norgestrel (NOG); and estradiol (E2), and monomeric and dimeric SHBG. Protein-ligand complexes, involving dimeric SHBG saturated with two ligands on each subunit, reproduce the experimental affinity tendency and allow the observation that dimerization exerts disparate effects on binding affinity, characteristic of negative cooperativity for E2, DHT, and NOG, whereas 3β-17α-diol and 3β-17β-diol lack allostery.


Thermal perception and lung function: a panel study in young adults with exercise under high outdoor temperature.
It has been observed that high temperature exposure is associated with a reduction in lung function and some possible biological mechanisms have been suggested. However, it is unclear if thermal perception plays a role in the association. From September 3rd to 15th, 2018, in Guangzhou, China, we repeatedly measured daily thermal perception and lung function among 126 participants with outdoor military training. We performed a linear mixed model and stratified analyses by the origin of students, gender, and the training period to evaluate the effects of thermal perception on lung function. A total of 399 measurements were collected. Per vote increase in thermal sensation vote towards the "hot" direction was associated with a - 0.04 L (95% CI: - 0.08 to - 0.01) decrease in forced vital capacity (FVC), and - 0.04 L (95% CI: - 0.08 to - 0.01) decrease in forced expiratory volume in 1 s (FEV1). Per grade increase towards the "very uncomfortable" direction for thermal comfort vote was associated with an increased percentage of forced expiratory volume in 1 s (FEV1%) by 1.52% (95% CI: 0.18 to 2.86). For thermal preference, with preferred cooler vote increased by one level, FVC and FEV1 decreased by - 0.05 L/s (95% CI: - 0.08 to - 0.02) and - 0.05L/s (95% CI: - 0.08 to - 0.02), respectively. The effects of thermal perception on lung function were stronger among non-local and in the first week of training. Our study suggests that in the same high-temperature environment, thermal perception is associated with lung function, even in healthy adults.


Verbal fluency as a quick and simple tool to help in deciding when to refer patients with a possible brain tumour.
Patients with brain tumours often present with non-specific symptoms. Correctly identifying who to prioritise for urgent brain imaging is challenging. Brain tumours are amongst the commonest cancers diagnosed as an emergency presentation. A verbal fluency task (VFT) is a rapid triage test affected by disorders of executive function, language and processing speed. We tested whether a VFT could support identification of patients with a brain tumour. This proof-of-concept study examined whether a VFT can help differentiate patients with a brain tumour from those with similar symptoms (i.e. headache) without a brain tumour. Two patient populations were recruited, (a) patients with known brain tumour, and (b) patients with headache referred for Direct-Access Computed-Tomography (DACT) from primary care with a suspicion of a brain tumour. Semantic and phonemic verbal fluency data were collected prospectively. 180 brain tumour patients and 90 DACT patients were recruited. Semantic verbal fluency score was significantly worse for patients with a brain tumour than those without (P < 0.001), whether comparing patients with headache, or patients without headache. Phonemic fluency showed a similar but weaker difference. Raw and incidence-weighted positive and negative predictive values were calculated. We have demonstrated the potential role of adding semantic VFT score performance into clinical decision making to support triage of patients for urgent brain imaging. A relatively small improvement in the true positive rate in patients referred for DACT has the potential to increase the timeliness and efficiency of diagnosis and improve patient outcomes.


The epidemiology of lymphoma in Jordan: A nationwide population study of 4189 cases according to World Health Organization classification system.
Lymphoma is a common human cancer that shows a variable geographic incidence worldwide. It is the fourth most common cancer in Jordan. Systemic reports of descriptive epidemiology on lymphoma from the Middle East are limited. A nationwide multi-institutional retrospective study was conducted covering all major hospitals and laboratories that provide diagnostic services. We collected data on all cases diagnosed with lymphoma between 2014 and 2019. The included variables were patients' age, gender, anatomic site, and the histologic type according to the World Health Organization classification system. A total of 4189 cases were diagnosed with lymphoma. There was a statistically significant gender difference (p < .05), as 57.5% of patients were males. The peak incidence occurred at age 25-55 years. There were 1,652 (39%) cases of Hodgkin lymphoma (HL) and 2,537 (61%) of non-Hodgkin lymphoma (NHL), where nodular sclerosis (67%) and diffuse large B-cell lymphoma (53%) were the most common subtypes, respectively. The average age-adjusted incidence rates per 100,000 population were 8.01 for all lymphomas, 4.33 for NHL, and 3.16 for HL and all remained stable over the 6 years. HL is the most common lymphoma in Jordan, with a percentage higher than most of reported studies in Asian and Western countries. It also shows a unimodal distribution of age-specific incidence rates, with a single peak in young adults. The incidence rate of HL is higher than Eastern countries but comparable to the West. In contrast, NHL demonstrates a lower incidence rate than Western countries but a similar distribution of subtypes, as mature T/natural killer-cell lymphomas were rare.


Modified frailty index predicts high-risk patients for readmission after colorectal surgery for cancer.
Modified frailty index (mFI) has been proposed as a reliable tool in predicting postoperative outcomes after surgery. This study aims to evaluate whether mFI could be utilized to predict readmissions after colorectal resection for patients with cancer by using nationwide cohort. Patients undergoing elective abdominal colorectal resection for colorectal cancer were reviewed from the American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP) procedure-targeted database (2010-2012). A previously described mFI was calculated. Demographics, comorbidities, and 30-day postoperative complications were compared between patients who were readmitted or not after colorectal surgery. A total of 7337 patients were identified with a mean age of 65.8(±13.6) years. Eight hundred seventy-one (11.8%) patients were readmitted at least once within 30 days. Age, gender, BMI, and other comorbidities were comparable between the groups. O approach, current smoking, mFI(>3/11), disseminating cancer, bleeding disorder and longer operative time were found to independently associated with readmission. An 11-point modified frailty index as measured in NSQIP correlates with readmissions after colorectal resection in patients with colon and rectal cancer.


Structure of sorting nexin 11 (SNX11) reveals a novel extended phox homology (PX) domain critical for inhibition of SNX10-induced vacuolation.
Sorting nexins are phox homology (PX) domain-containing proteins involved in diverse intracellular endosomal trafficking pathways. The PX domain binds to certain phosphatidylinositols and is recruited to vesicles rich in these lipids. The structure of the PX domain is highly conserved, containing a three-stranded β-sheet, followed by three α-helices. Here, we report the crystal structures of truncated human SNX11 (sorting nexin 11). The structures reveal that SNX11 contains a novel PX domain, hereby named the extended PX (PXe) domain, with two additional α-helices at the C terminus. We demonstrate that these α-helices are indispensible for the in vitro functions of SNX11. We propose that this PXe domain is present in SNX10 and is responsible for the vacuolation activity of SNX10. Thus, this novel PXe domain constitutes a structurally and functionally important PX domain subfamily.


Association of frabin with specific actin and membrane structures.
Frabin is an actin filament (F-actin)-binding protein with GDP/GTP exchange activity specific for Cdc42 small G protein. Expression of frabin forms filopodia-like microspikes through the direct activation of Cdc42, and lamellipodia through indirect activation of Rac small G protein. Frabin consists of the F-actin-binding domain (FAB), the Dbl homology domain (DH), the first pleckstrin homology domain (PH1), the FYVE-finger domain (FYVE), the second PH domain (PH2) from the N-terminus in this order. Although DH and PH1 show exchange activity, FAB, in addition to DH and PH1, is required for the formation of microspikes, whereas FYVE and PH2, in addition to DH and PH1, are required for the formation of lamellipodia. Various truncated mutants of frabin were co-expressed with a dominant active mutant (DA) of Cdc42, Rac1DA, or full-length frabin in L fibroblasts. FAB was recruited to the Cdc42DA-formed filopodia-like microspikes. FAB and a fragment containing DH, PH1, FYVE and PH2 were recruited to the Rac1DA-formed membrane ruffles. Furthermore, each of these fragments served as a dominant negative mutant of frabin when co-expressed with full-length frabin, and inhibited the full-length frabin-formed morphological changes. These results suggest that frabin recognizes a specific actin structure(s) through FAB and a specific membrane structure(s) through FAB and the region containing DH, PH1, FYVE and PH2. It is likely that frabin associates with the specific actin and membrane structures and activates Cdc42 and Rac in the vicinity of these structures, eventually leading to morphological changes.


The X-ray crystallographic study of long chain left handed Z-DNA and polyamine complex and structural chemistry study of effection by polyamine for the Z-DNA.
We cleared the stabilization mechanism of Z-DNA by using the molecular dynamics calculation and the grid map simulated annealing docking method. Then, we clarified Z-DNA was stabilized by the metal counter ions whose electrical charge and the number of electrons are too many and by polyamine to go into the minor groove. We succeeded in stabilizing Z-DNA of the long chain by using these methods, and succeeded in the crystallization and the data collection up to 1.7 A resolution.


Somatostatin modulates dopamine release in rat retina.
The aim of the present study was to determine the possible role of somatostatin as a modulator of dopamine release in rat retina. Basal release of dopamine, and how this is influenced by somatostatin receptor (sst) selective ligands, was examined ex vivo in rat retinal explants. Dopamine levels were quantified by high-pressure liquid chromatography (HPLC) with electrochemical detection. Basal levels of dopamine were measured over 120 min of tissue incubation and found to be 1.17+/-0.35 ng/ml. Somatostatin (10(-6), 10(-5), 10(-4)M) increased dopamine levels in a concentration-dependent manner, while the sst(2) antagonist CYN154806 (10(-4)M) reversed its actions. BIM23014 (sst(2) agonist) increased dopamine levels in a statistically significant manner only at the concentration of 10(-5)M. The sst(1) agonist L797.591 (10(-5), 10(-4)M) also increased dopamine levels, while activation of the sst(3) receptor (sst(3) agonist, L796.778, 10(-4)M) had no effect. These data substantiate a neuromodulatory role for sst(1) and sst(2) somatostatin receptors in the retina and show for the first time somatostatin's influence on dopamine release.


The production of antibodies to vitamin B-12.
The conjugation of vitamin B-12 to HSA by means of a combined ethylcarbodiimide N-hydroxysuccinimide conjugation method at pH 5.5 is described. It gave a ratio of 2.7 mol vitamin B-12/mol HSA. Five rabbits were immunized with this conjugate emulsified in Freund's complete adjuvant. After one booster injection high titre antisera (1 : 450 000--1 : 1 500 00) to vitamin B-12 were collected.


Monitoring of bentonite pore water with a probe based on solid-state microsensors.
Repositories for the disposal of radioactive waste generally rely on a multi-barrier system to isolate the waste from the biosphere. This multi-barrier system typically comprises the natural geological barrier provided by the repository host rock and its surroundings and an engineered barrier system (EBS). Bentonite is being studied as an appropriated porous material for an EBS to prevent or delay the release and transport of radionuclides towards biosphere. The study of pore water chemistry within bentonite barriers will permit to understand the transport phenomena of radionuclides and obtain a database of the bentonite-water interaction processes. In this work, the measurement of some chemical parameters in bentonite pore water using solid-state microsensors is proposed. Those sensors are well suited for this application since in situ measurements are feasible and they are robust enough for the long periods of time that monitoring is needed in an EBS. A probe containing an ISFET (ion sensitive field effect transistor) for measuring pH, and platinum microelectrodes for measuring conductivity and redox potential was developed, together with the required instrumentation, to study the chemical changes in a test cell with compacted bentonite. Response features of the sensors' probe and instrumentation performance in synthetic samples with compositions similar to those present in bentonite barriers are reported. Measurements of sensors stability in a test cell are also presented.


Expression of estrogen related proteins in hormone refractory prostate cancer: association with tumor progression.
Despite increasing evidence that estrogen signaling has a key role in prostate cancer development and progression, few studies have focused on the estrogen pathway in the transition from hormone sensitive to hormone refractory tumors. We investigated the expression of proteins related to androgen and estrogen metabolism in paired prostate cancer samples collected before androgen deprivation therapy and after hormonal relapse. The study included 55 patients treated for prostate cancer only with androgen deprivation therapy and in whom tissue was available before treatment induction and after recurrence. Immunohistochemistry was performed using tissue microarray with antibodies directed against androgen receptor, phosphorylated androgen receptor, estrogen receptor α, estrogen receptor β, 5α-reductase 1 and 2, aromatase, BCAR1 and the proliferation marker Ki67. Compared to hormone sensitive samples, tissues collected after hormonal relapse were characterized by increased expression of Ki67, androgen receptor, phosphorylated androgen receptor (p <0.001) and BCAR (p = 0.03), and by lower staining for 5α-reductase 2 (p = 0.002), estrogen receptor β (p = 0.016) and aromatase (p <0.001). Shorter time to hormonal relapse was associated with high expression of aromatase and BCAR1 on diagnostic biopsy, together with low staining for estrogen receptor α in stromal cells. Overall survival was significantly shorter when tissues collected after relapse showed a high proliferation index and low estrogen receptor α expression. Results revealed dysregulation of proteins involved in androgen pathways, and in estrogen synthesis and signaling during the development of hormone refractory prostate cancer.


The influence of localised size reorganisation on short-duration bidispersed granular flows.
We investigate experimentally the runout resulting from the collapse of a granular column containing two particle species that differ in size only. The experimental configuration is strictly two-dimensional (only one particle per width of the experimental tank) and we explore both the role of the initial arrangement and proportion of the two particle sizes in the column, using high-speed videography, and by determining the centres of mass of the big and small particles in the initial column and the final deposit. The duration of the experiment is sufficiently short that large-scale segregation does not occur, however, we find a clear dependence of runout on both initial mixture arrangement and proportion for all conditions. We investigated this observation through detailed analysis of the flow front motion, and identify a characteristic "stopping" phase when dissipation dominates, and we apply a shallow layer model at the flow front to show how the initial mixture arrangement and proportion influence the effective coefficient of friction during emplacement. We find that a bidispersed mixture can induce a larger friction on emplacement than a monodispersed mixture, and the highest coefficient of friction was found for a well-mixed initial arrangement of particles at the proportion that shows maximum horizontal spreading of the flow. These observations suggest that downwards percolation of fine particles takes place at the front of the collapsing column, and so localised size segregation processes at the flow front can control flow mobility. This effect is likely to be important in controlling the mobility of large geophysical flows that occur on finite time scales, and whose deposits typically show granular segregation at the front and edges but not throughout the entire deposit.


Weissella paramesenteroides WpK4 plays an immunobiotic role in gut-brain axis, reducing gut permeability, anxiety-like and depressive-like behaviors in murine models of colitis and chronic stress.
The relationship between inflammatory bowel disease (IBD) and mood disorders is complex and involves overlapping metabolic pathways, which may determine comorbidity. Several studies have been shown that this comorbidity could worsen IBD clinical course. The treatment of ulcerative colitis is complex, and involves traditional therapy to promote the function of epithelial barrier, reducing exacerbated inflammatory responses. Recently, it has been shown that some probiotic strains could modulate gut-brain axis, reducing depressive and anxiety scores in humans, including IBD patients. Accordingly, this study aimed to evaluate the role of Weissella paramesenteroides WpK4 in murine models of ulcerative colitis and chronic stress. It was observed that bacterium ingestion improved health of colitis mice, reducing intestinal permeability, besides improving colon histopathological appearance. In stressed mice, bacterial consumption was associated with a reduced anxiety-like and depressive-like behaviors. In both assays, the beneficial role of W. paramesenteroides WpK4 was related to its immunomodulatory feature. It is possible to state that W. paramesenteroides WpK4 exerted their beneficial roles in gut-brain axis through their immunomodulatory effects with consequences in several metabolic pathways related to intestinal permeability and hippocampal physiology.


[RNA synthesis in the polymorphonuclear leukocytes of a suppurative wound].
RNA synthesis in biopsy specimens of burn or traumatic wounds and in blood leukocytes was investigated in 11 patients by electron microscopic radioautography. Experiments with leukocytes were carried out in two variants: 1) incubation with 3H-uridine; 2) incubation with 3H-uridine in the presence of Staphylococcus aureus of Staphylococcus epidermidis suspensions. It was detected that neutrophils incubated without bacteria did not contain the label. Neutrophils incubated with bacteria and phagocytizing them could incorporate 3H-uridine. The label was insignificant in this case and the number of labeled cells did not exceed 18%. The majority of neutrophils (70-90%) in pus accumulations from burn and traumatic wounds contained the label in a higher concentration than neutrophils that phagocytize bacteria in test-tubes.


On the Teaching of Science, Technology and International Affairs.
Despite the ubiquity and critical importance of science and technology in international affairs, their role receives insufficient attention in traditional international relations curricula. There is little literature on how the relations between science, technology, economics, politics, law and culture should be taught in an international context. Since it is impossible even for scientists to master all the branches of natural science and engineering that affect public policy, the learning goals of students whose primary training is in the social sciences should be to get some grounding in the natural sciences or engineering, to master basic policy skills, to understand the basic concepts that link science and technology to their broader context, and to gain a respect for the scientific and technological dimensions of the broader issues they are addressing. They also need to cultivate a fearless determination to master what they need to know in order to address policy issues, an open-minded but skeptical attitude towards the views of dueling experts, regardless of whether they agree with their politics, and (for American students) a world-view that goes beyond a strictly U.S. perspective on international events. The Georgetown University program in Science, Technology and International Affairs (STIA) is a unique, multi-disciplinary undergraduate liberal arts program that embodies this approach and could be an example that other institutions of higher learning might adapt to their own requirements.


Effects of Coke Deposits on the Catalytic Performance of Large Zeolite H-ZSM-5 Crystals during Alcohol-to-Hydrocarbon Reactions as Investigated by a Combination of Optical Spectroscopy and Microscopy.
The catalytic activity of large zeolite H-ZSM-5 crystals in methanol (MTO) and ethanol-to-olefins (ETO) conversions was investigated and, using operando UV/Vis measurements, the catalytic activity and deactivation was correlated with the formation of coke. These findings were related to in situ single crystal UV/Vis and confocal fluorescence micro-spectroscopy, allowing the observation of the spatiotemporal formation of intermediates and coke species during the MTO and ETO conversions. It was observed that rapid deactivation at elevated temperatures was due to the fast formation of aromatics at the periphery of the H-ZSM-5 crystals, which are transformed into more poly-aromatic coke species at the external surface, preventing the diffusion of reactants and products into and out of the H-ZSM-5 crystal. Furthermore, we were able to correlate the operando UV/Vis spectroscopy results observed during catalytic testing with the single crystal in situ results.


Association of physical activity metrics with indicators of cardiovascular function and control in children with and without type 1 diabetes.
Little is known about the role of physical activity accumulation in cardiovascular disease risk for children with type 1 diabetes. Improved insight to identify factors of influence in key health outcomes could be provided by considering the entire physical activity profile. Pulse wave velocity (PWV), augmentation index and heart rate variability (HRV) were assessed cross-sectionally in children with (n = 29, 12.1 ± 2.1 years) and without (n = 19, 12.1 ± 2.1 years) type 1 diabetes. Time spent sedentary and in each physical activity intensity, intensity gradient and average acceleration were derived from seven consecutive days of monitoring with wrist-worn accelerometry. Comparison between groups and influence of physical activity accumulation on cardiovascular metrics were explored with linear mixed models. Diabetic children demonstrated a higher PWV and a greater volume of light physical activity (p < 0.01), a more negative intensity gradient (p < 0.01), a lower average acceleration and less time in bouted moderate-to-vigorous physical activity (MVPA; p < 0.05). Overall, intensity gradient was strongly correlated with average acceleration, MVPA and bouted MVPA (r2 = 0.89, r2 = 0.80, r2 = 0.79, respectively; all p < 0.05), while average acceleration was correlated with MVPA and bouted MVPA (r2 = 0.85, r2 = 0.83, respectively; p < 0.05). Accounting for disease status, intensity gradient and average acceleration were significant predictors of HRV indices (p < 0.05) and PWV (p < 0.01, p < 0.05, respectively). Overall, MVPA was most associated with central stiffness, highlighting the importance of meeting activity guidelines. Diabetic children demonstrated poorer cardiovascular health than their counterparts, likely attributable to a lower intensity and physical activity volume, identifying physical activity intensity as a key target for future interventions.


Repair of a mycotic aneurysm of the infrarenal aorta in a patient with HIV, using a Palmaz stent and autologous femoral vein graft.
With the widespread use of endovascular surgery, aneurysms can be managed selectively with the use of stent grafts. Standard treatment of mycotic aneurysms usually requires resection and extraanatomic bypass. Although stent graft repair of a mycotic femoral aneurysm with autologous graft has been reported, we present a case of an infrarenal mycotic aneurysm in a patient with the human immunodeficiency virus (HIV) that was successfully treated with a novel endovascular approach.


The intracranial topography of the P3 event-related potential elicited during auditory oddball.
In order to isolate the anatomical locus of neural activity primarily responsible for generating the scalp-recorded P3 (or P300), the topography of event-related potentials (ERPs) elicited during an auditory oddball task was compared between medial-to-lateral aspects of the frontal, parietal, and temporal lobes in 10 epileptic patients undergoing stereoelectroencephalography for seizure localization. Evidence of local ERP generation was obtained from each of these areas. Small amplitude P3-type potentials were sometimes observed to invert polarity across recording contacts in the frontal lobe. Large amplitude positive polarity P3-type components were observed in the lateral neocortex of the inferior parietal lobule (IPL), that rapidly attenuated in amplitude at more anterior, posterior, superior, inferior, and medial recording contacts. Large amplitude polarity inverting P3-type components were also observed to be highly localized to hippocampal contacts of temporal lobe electrodes. These data are discussed in the context of other recent studies of lesion effects, scalp topography, and intracranial recordings, and it is concluded that activity generated in the IPL is likely to make the major contribution to the scalp-recorded P3, with smaller contributions from these other sources. Finally, salient topographical differences between the intracranial distribution of the P3 and those of the N2 (or N200) and slow wave (SW) suggest that the generators of these components are not identical.


Spectrofluorimetric determination of fluvoxamine in dosage forms and plasma via derivatization with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole.
A highly sensitive and simple spectrofluorimetric method has been developed and validated for the determination of the antidepressant fluvoxamine (FXM) in its dosage forms and plasma. The method was based on nucleophilic substitution reaction of FXM with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole in an alkaline medium (pH 8) to form a highly fluorescent derivative that was measured at 535 nm after excitation at 470 nm. The factors affecting the reaction was carefully studied and optimized. The kinetics of the reaction was investigated, and the reaction mechanism was presented. Under the optimized conditions, linear relationship with good correlation coefficient (0.9995) was found between the fluorescence intensity and FXM concentration in the range of 65-800 ng ml(-1). The limits of detection and quantitation for the method were 21 and 64 ng ml(-1), respectively. The precision of the method was satisfactory; the values of relative standard deviations did not exceed 2.17%. The proposed method was successfully applied to the determination of FXM in its pharmaceutical tablets with good accuracy; the recovery values were 97.8-101.4 +/- 1.08-2.75%. The results obtained by the proposed method were comparable with those obtained by the official method. The high sensitivity of the method allowed its successful application to the analysis of FXM in spiked human plasma. The proposed method is superior to the previously reported spectrofluorimetric method for determination of FXM in terms of its simplicity. The proposed method is practical and valuable for its routine application in quality control and clinical laboratories for analysis of FXM.


Intra-amniotic injection of IL-1 induces inflammation and maturation in fetal sheep lung.
Antenatal inflammation may be an important triggering event in the pathogenesis of bronchopulmonary dysplasia but may also accelerate fetal lung maturation. We examined the effects of intra-amniotic (IA) interleukin (IL)-1 alpha and IL-1 beta on maturation of the fetal sheep lung. These cytokine effects were compared with IA endotoxin, a potent proinflammatory stimulus that accelerated lung maturation. Date-bred ewes received 15 or 150 microg recombinant ovine IL-1 alpha or IL-1 beta or 10 mg Escherichia coli endotoxin by IA injection at 118 days gestation (term = 150 days), and fetuses were delivered at 125 days. IL-1 alpha and IL-1 beta improved lung function and increased alveolar saturated phosphatidylcholine (Sat PC) and surfactant protein mRNA expression at the higher dose. The maturation response to IL-1 alpha was greater than that to IL-1 beta, which was similar to endotoxin response. Inflammation was also more pronounced after IL-1 alpha treatment. Only endotoxin animals had residual inflammation of the fetal membranes at 7 days. Lung compliance, lung volume, and alveolar Sat PC were positively correlated with residual alveolar wash leukocyte numbers 7 days after IL-1 treatment, suggesting a link between lung inflammation and maturation.


Release of TNF-alpha from lipopolysaccharide (LPS)-stimulated Kupffer cells in serum- and nutrient-free medium.
In monocytes/macrophages LPS stimulation occurs by the binding of LPS and the serum component LPS-binding protein (LBP) to CD14. This study was conducted to investigate whether this mechanism also occurs in Kupffer cells. Rat Kupffer cells were stimulated for up to 8 h by LPS (0, 100 ng/ml, 10 microg/ml) in RPMI medium or in nutrient-free Krebs-Henseleit (KH) buffer. Some incubations were performed without serum, while in others serum was provided. TNF-alpha concentrations of the supernatants were measured by ELISA. LPS stimulation of Kupffer cells yielded the following results. In KH without any additives a considerable amount of TNF-alpha was released. Incubation in RPMI without serum caused twice as much TNF-alpha to be released as when KH was used. The addition of autologous serum to RPMI did not increase TNF-alpha response. These results provide evidence that a substantial part of TNF-alpha release by LPS-stimulated Kupffer cells occurs in a serum- and thus LBP-independent way.


Symptoms and signs in pulmonary medicine: old observations and new interpretations.
Andre Gide wrote, "Everything has been said before, but since nobody listens we have to keep going back and beginning all over again." To a certain extent, that statement applies to the importance of accurate and systematic history taking and physical examination in clinical practice. Although we are trained in habits of comprehension and accuracy in history taking and examination of patients, periodic reminders are required to develop a diagnostic framework based on observation (inspection), palpation, percussion, and auscultation. This monograph reemphasizes the method to be pursued in the treatment of a patient with pulmonary symptoms. It consists of three parts: (1) questioning the patient about his or her medical history; (2) performing the physical examination of the respiratory system; and (3) examining the extrapulmonary signs and symptoms. Once a strong clinical framework has been constructed, its further development and refinement depend on the clinician's experience, power of observation, and systematic reading of the medical literature. Good physicians must continue to learn throughout their careers; this is the most essential element of a physician's development. Be patient, however; as Cowper said, "Knowledge, to become wisdom, needs experience."


Treatment of carpal tunnel syndrome. Comparative study with and without epineurolysis.
Three series of cases of Carpal Tunnel Syndrome have been studied. One series was treated a different, respected surgical team, the second series by the author by decompression alone, and the third series by additional intraneural decompression using the microscope. The authors found very little difference in the results, apart from marginally better results in the presence of thenar muscle involvement.


Leveraging AI Advances and Online Tools for Structure-Based Variant Analysis.
Understanding how a gene variant affects protein function is important in life science, as it helps explain traits or dysfunctions in organisms. In a clinical setting, this understanding makes it possible to improve and personalize patient care. Bioinformatic tools often only assign a pathogenicity score, rather than providing information about the molecular basis for phenotypes. Experimental testing can furnish this information, but this is slow and costly and requires expertise and equipment not available in a clinical setting. Conversely, mapping a gene variant onto the three-dimensional (3D) protein structure provides a fast molecular assessment free of charge. Before 2021, this type of analysis was severely limited by the availability of experimentally determined 3D protein structures. Advances in artificial intelligence algorithms now allow confident prediction of protein structural features from sequence alone. The aim of the protocols presented here is to enable non-experts to use databases and online tools to investigate the molecular effect of a genetic variant. The Basic Protocol relies only on the online resources AlphaFold, Protein Structure Database, and UniProt. Alternate Protocols document the usage of the Protein Data Bank, SWISS-MODEL, ColabFold, and PyMOL for structure-based variant analysis. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: 3D Mapping based on UniProt and AlphaFold Alternate Protocol 1: Using experimental models from the PDB Alternate Protocol 2: Using information from homology modeling with SWISS-MODEL Alternate Protocol 3: Predicting 3D structures with ColabFold Alternate Protocol 4: Structure visualization and analysis with PyMOL.


To what extent are we really free from airborne microplastics?
Quality assurance and quality control (QA&QC) procedures are vital for ensuring data reliability, but little is known about the use of such procedures in reducing airborne microplastic (MP) contamination. To address this issue, we tried to determine the efficiency of two common methods (washing and ashing experimental glassware) for removing airborne MPs and identified airborne contamination during MP analytical procedure. The results showed the removal efficiencies of washing and ashing were an average of 88%-98% and 100%, respectively, indicating that both methods could eliminate most of the spiked airborne MPs with no significant difference noted between the two methods. Although rigorous measures were taken to prevent contamination from ambient air, trace amounts of airborne MPs were still detected, which is an issue that has not been adequately investigated in previous studies. All of the procedural contaminants detected in this study were fibrous. Approximately 88% of these fibers were cotton-like (cotton, cellulose, and cellophane) fibers, and 13% of them were plastic. Surprisingly, cotton-like fibers and MPs had a similar size distribution, suggesting that they may have undergone a similar weathering process. In the end, to cope with inevitable airborne contamination, several measures were proposed for further research. Such measures will provide the necessary methodological assistance for accurate quantification of MP pollution in the field.


Identification of key signaling pathways induced by SARS-CoV2 that underlie thrombosis and vascular injury in COVID-19 patients.
The SARS-CoV-2 pandemic has led to hundreds of thousands of deaths and billions of dollars in economic damage. The immune response elicited from this virus is poorly understood. An alarming number of cases have arisen where COVID-19 patients develop complications on top of the symptoms already associated with SARS, such as thrombosis, injuries of vascular system, kidney, and liver, as well as Kawasaki disease. In this review, a bioinformatics approach was used to elucidate the immune response triggered by SARS-CoV-2 infection in primary human lung epithelial and transformed human lung alveolar. Additionally, examined the potential mechanism behind several complications that have been associated with COVID-19 and determined that a specific cytokine storm is leading to excessive neutrophil recruitment. These neutrophils are directly leading to thrombosis, organ damage, and complement activation via neutrophil extracellular trap release.


Wasp uses venom cocktail to manipulate the behavior of its cockroach prey.
The sting of the parasitoid wasp Ampulex compressa is unusual, as it induces a transient paralysis of the front legs followed by grooming behavior and then by a long-term hypokinesia of its cockroach prey. Because the wasp's goal is to provide a living meal for its newborn larva, the behavioral changes in the prey are brought about by manipulating the host behavior in a way beneficial to the wasp and its offspring. To this end, the wasp injects its venom cocktail with two consecutive stings directly into the host's central nervous system. The first sting in the thorax causes a transient front leg paralysis lasting a few minutes. This paralysis is due to the presence of a venom component that induces a postsynaptic block of central cholinergic synaptic transmission. Following the head sting, dopamine identified in the venom appears to induce 30 min of intense grooming. During the long-term hypokinesia that follows the grooming, specific behaviors of the prey are inhibited while others are unaffected. We propose that the venom represses the activity of head ganglia neurons thereby removing the descending excitatory drive to the thoracic neurons.


Analytical methods for amatoxins: A comprehensive review.
Amatoxins are toxic bicyclic octapeptides found in certain wild mushroom species, particularly Amanita phalloides. These mushrooms contain predominantly α- and β-amanitin, which can lead to severe health risks for humans and animals if ingested. Rapid and accurate identification of these toxins in mushroom and biological samples is crucial for diagnosing and treating mushroom poisoning. Analytical methods for the determination of amatoxins are critical to ensure food safety and prompt medical treatment. This review provides a comprehensive overview of the research literature on the determination of amatoxins in clinical specimens, biological and mushroom samples. We discuss the physicochemical properties of toxins, highlighting their influence on the choice of the analytical method and the importance of sample preparation, particularly solid-phase extraction with cartridges. Chromatographic methods are emphasised with a focus on liquid chromatography coupled to mass spectrometry as one of the most relevant analytical method for the determination of amatoxins in complex matrices. Furthermore, current trends and future perspectives in amatoxin detection are also suggested.


Validity of the Patient Health Questionnaire-9 for depression screening and diagnosis in East Africa.
Depression is often underdiagnosed and undertreated in primary care settings, particularly in developing countries. This is, in part, due to challenges resulting from lack of skilled mental health workers, stigma associated with mental illness, and lack of cross-culturally validated screening instruments. We conducted this study to evaluate the reliability and validity of the Patient Health Questionnaire-9 (PHQ-9) as a screen for diagnosing major depressive disorder among adults in Ethiopia, the second most populous country in sub-Saharan Africa. A total of 926 adults attending outpatient departments in a major referral hospital in Ethiopia participated in this study. We assessed criterion validity and performance characteristics against an independent, blinded, and psychiatrist administered semi-structured Schedules for Clinical Assessment in Neuropsychiatry (SCAN) interview. Overall, the PHQ-9 items showed good internal (Cronbach's alpha=0.81) and test re-test reliability (intraclass correlation coefficient=0.92). A factor analysis confirmed a one-factor structure. Receiver Operating Characteristics (ROC) analysis showed that a PHQ-9 threshold score of 10 offered optimal discriminatory power with respect to diagnosis of major depressive disorder via the clinical interview (sensitivity=86% and specificity=67%). The PHQ-9 appears to be a reliable and valid instrument that may be used to diagnose major depressive disorders among Ethiopian adults.


Concomitant Administration of a Histamine2 Receptor Antagonist and Proton Pump Inhibitor Enhances Gastric Acid Suppression.
Because it has been hypothesized that histamine2 receptor antagonists (H2 RAs) might interfere with the action of proton pump inhibitors (PPIs) when the drugs are given concomitantly, we sought to compare the pharmacodynamic effects of simultaneous administration of a PPI and an H2 RA with the effects of each drug administered alone. Prospective, randomized, double-blind, three-way crossover study. Esophageal motility laboratory at a large teaching hospital. Twenty-one healthy volunteers. Subjects were randomized to one of three treatment arms: an H2 RA (ranitidine 300 mg) plus placebo, a PPI (omeprazole 40 mg) plus placebo, or ranitidine 300 mg plus omeprazole 40 mg, all given once/day at 8 a.m., 30 minutes before a standard breakfast, for 1 week. The subjects then received the other two treatments, with each treatment period separated by a 1-week washout period. The primary outcome was length of time that the gastric pH remained higher than 4. Secondary outcomes were median gastric pH higher than 4 and percentage of time that the gastric pH remained higher than 4. On day 7, ambulatory intragastric pH was recorded over an 8-hour period in each treatment arm. The combination of ranitidine and omeprazole resulted in a significantly longer time that the gastric pH remained higher than 4 (median 410.5 min [interquartile range (IQR) 298.5-454.25 min]) versus either omeprazole alone (median 356.7 min [IQR 254.9-419.2 min], p=0.023) or ranitidine alone (134.1 min [IQR 99.9-302.5 min], p<0.0001). Median gastric pH was also significantly higher when omeprazole and ranitidine were given in combination (pH 5.92 [IQR 4.75-6.46]) than either omeprazole alone (pH 4.88 [IQR 4.27-6.11], p=0.001) or ranitidine alone (pH 2.31 [IQR 2.04-5.27], p=0.0003). Likewise, the percentage of time that the gastric pH remained higher than 4 was significantly higher when omeprazole and ranitidine were given in combination (median 85.52%) than either omeprazole alone (74.31%, p=0.027) or ranitidine alone (27.94%, p=0.0002). When a PPI and H2 RA were administered concomitantly 30 minutes before breakfast, the H2 RA did not decrease the acid suppressive ability of the PPI; rather, it improved gastric acid control. Thus these results failed to support the initial hypothesis of this study. Further prospective studies are needed to test these findings in patients with gastroesophageal reflux disease as well as those with erosive esophagitis.


[Evaluation of non-neurogenic overactive bladder syndrome].
International guidelines exist regarding the initial assessment of patients suffering from overactive bladder (OAB). To synthesize current knowledge on the evaluation of OAB. A systematic literature review based on Pubmed, Embase, Google Scholar was conducted in June 2020. An accurate taking of history and assessment of lower urinary tract symptoms are the first steps of the evaluation of patients with OAB. In addition, the search of risk factors for OAB, the exclusion of urological causes which can be responsible for urgency and the identification of therapeutic contra-indications are essential. The clinical examination and a 3-to 7-day bladder diary are part of the initial assessment. Self-questionnaires validated in French measure patients' bother and the impact on quality of life. The urine strip or culture eliminates a urinary tract infection. Urinary cytology and cystoscopy investigate a bladder tumor in case of risk factor. The post-void residual volume has to be measured in case of voiding symptoms. Urodynamics and imaging are not first-line tests. The understanding of the initial evaluation of OAB is essential to introduce treatments adapted to patients' bother.


[False aneurysm of the external iliac artery revealing Behçet's disease].
Behçet's disease is a vasculitis of unknown etiology. Vascular involvement is rare, but may be inaugural in many cases. We report a case of Behçet's disease revealed by a symptomatic pseudo-aneurysm of the external iliac artery. The symptomatology was non-specific. Ultrasound Doppler and computed tomographic angiography were essential for diagnosis. The pseudo-aneurysm was managed by endovascular treatment. Corticosteroid and immunosuppressant therapy were administrated after surgery.


Penaeus monodon Tudor staphylococcal nuclease preferentially interacts with N-terminal domain of Argonaute-1.
RNA interference (RNAi) plays a crucial role as an antiviral defense in several organisms including plants and invertebrates. An understanding of RNAi machineries especially protein components of the RNA-induced silencing complex (RISC) is essential for prior to applying RNAi as a tool for viral protective immunity in shrimp. Tudor staphylococcal nuclease (TSN) is an evolutionarily conserved protein and is one of the RISC components. In previous study, suppression of Penaeus monodon TSN (PmTSN) by double-stranded RNA (dsRNA) resulted in decreasing dsRNA-mediated gene silencing activity. To elucidate the functional significance of PmTSN in shrimp RNAi pathway, interactions between PmTSN and three Argonaute proteins (PmAgo) were characterized by yeast two-hybrid and in vitro pull-down assays. The results demonstrated that PmTSN interacted with PmAgo1, but not with PmAgo2 or PmAgo3. The interaction between PmAgo and PmTSN was mediated through the N-terminal domain of PmAgo1 and the SN1-2 domains of PmTSN. Analysis of the nuclease activity of the recombinant PmTSN indicated that PmTSN possessed calcium-dependent nuclease activity specific to single-stranded RNA (ssRNA), but not dsRNA and DNA. Knockdown of PmAgo1 and PmTSN diminished the ability of dsRNA-Rab7 to knockdown PmRab7 expression, indicating the involvement of PmAgo1 and PmTSN in shrimp RNAi pathway. Taken together, the results imply that PmTSN is one of the components of PmAgo1-RISC, thus providing new insights in the RNAi-based mechanism in shrimp.