Plasma Fluorescent Oxidation Products and Bone Mineral Density Among Male Veterans: A Cross-Sectional Study.
In vitro and vivo studies indicate that oxidative stress contributes to bone loss. Fluorescent oxidation products (FlOPs) are novel biomarkers of oxidative stress; they reflect global oxidative damage of lipids, proteins, carbohydrates, and DNA. However, whether FlOPs are associated with bone mineral density (BMD) is still unclear. In the present study, we examined the association between FlOPs and BMD among male veterans. This cross-sectional study was conducted among participants recruited from the Department of Medical Examination, The Second Hospital of Jilin University in Jilin, China. We identified male veterans who were at least 50 y old between June and October of 2019. Plasma FlOPs were measured with a fluorescent microplate reader (excitation/emission wavelength: 320/420 nm). BMD were measured by dual-energy X-ray absorptiometry (DXA). The association between FlOPs and BMD was tested by multivariable linear regression models. A total of 164 male veterans were enrolled in the study, the average age was 56.6 y. After adjusting for covariates, veterans who had FlOP levels in the highest tertile had a statistically significant lower femoral neck (β = -0.044; p = 0.007) and total hip BMD (β = -0.045; p = 0.020) as compared to those with FlOP levels in the lowest tertile. Similar results were found when FlOPs were treated as a continuous variable (per 1-SD increase, β = -0.014 and p = 0.033 for femoral neck BMD; β = -0.016 and p = 0.047 for total hip BMD). Higher FlOP levels were associated with lower BMD among male veterans.


The endoplasmic reticulum is the site of cholesterol-induced cytotoxicity in macrophages.
Excess cellular cholesterol induces apoptosis in macrophages, an event likely to promote progression of atherosclerosis. The cellular mechanism of cholesterol-induced apoptosis is unknown but had previously been thought to involve the plasma membrane. Here we report that the unfolded protein response (UPR) in the endoplasmic reticulum is activated in cholesterol-loaded macrophages, resulting in expression of the cell death effector CHOP. Cholesterol loading depletes endoplasmic reticulum calcium stores, an event known to induce the UPR. Furthermore, endoplasmic reticulum calcium depletion, the UPR, caspase-3 activation and apoptosis are markedly inhibited by selective inhibition of cholesterol trafficking to the endoplasmic reticulum, and Chop-/- macrophages are protected from cholesterol-induced apoptosis. We propose that cholesterol trafficking to endoplasmic reticulum membranes, resulting in activation of the CHOP arm of the UPR, is the key signalling step in cholesterol-induced apoptosis in macrophages.


Can post-treatment free PSA ratio be used to predict adverse outcomes in recurrent prostate cancer?
To assess whether free PSA ratio (FPSAR) at biochemical recurrence (BCR) can predict metastasis, castrate-resistant prostate cancer (CRPC), and cancer-specific survival (CSS), following therapy for localised disease. A single-centre retrospective cohort study (NCT03927287) including a discovery cohort composed of patients with an FPSAR after radical prostatectomy (RP) or radiotherapy (RT) between 2000 and 2017. For validation, an independent Biobank cohort of patients with BCR after RP was tested. Using a defined FPSAR cut-off, the metastasis-free-survival (MFS), CRPC-free survival, and CSS were compared. Multivariable Cox models determined the association between post-treatment FPSAR, metastases, and CRPC. Overall, 822 patients (305 RP- and 363 RT-treated patients and 154 Biobank patients) were analysed. In the RP cohort, a total of 272/305 (89.1%) and 33/305 (10.9%) had a FPSAR test incidentally and reflexively, respectively. In the RT cohort, 155/363 (42.7%) and 208/263 (57.3%) had a FPSAR test incidentally and reflexively, respectively. However, in the prospective Biobank RP cohort, FPSAR testing was done on all samples of patients diagnosed with BCR. A FPSAR cut-off of 0.10 was determined using receiver operating characteristic analyses in both the RP and RT cohorts. A FPSAR of <0.10 resulted in longer median MFS (14.8 vs 9.3 years and 14.8 vs 13 years, respectively), and longer median CRPC-free survival (median not reached vs 9.9 years and 20.7 vs 13.8 years, respectively). Multivariable analyses showed that a FPSAR of ≥0.10 was associated with increased metastasis in the RP cohort (hazard ratio [HR] 1.915, 95% confidence interval [CI] 1.241-2.955) and RT cohort (HR 1.754, 95% CI 1.112-2.769), and increased CRPC in the RP cohort (HR 2.470, 95% CI 1.493-4.088). Findings were validated in the Biobank cohort. A post-treatment FPSAR of ≥0.10 is associated with more aggressive disease, suggesting a potentially novel role for this biomarker.


Regulation of endothelial nitric oxide synthase in cardiac remodeling.
Our previous study demonstrated that endothelial nitric oxide synthase (eNOS) gene serves as a candidate for modifiers of hypertrophic cardiomyopathy (HCM), which alters severity of HCM phenotypes. Herein, we sought to further elucidate the role of eNOS on cardiac myocyte hypertrophy and fibrosis, the major phenotypes of HCM. Male eNOS-deficient mice (eNOS-/-) and wild type control mice (eNOS+/+, C57B1/6 J) were used in this study. Myocyte size was analyzed in hematoxylin/eosin stained sections using an image analyzing system. Cardiac β-myosin heavy chain (β-MHC) and α-skeletal actin (α-SKA) levels, markers of myocyte hypertrophy were evaluated by Western blot. Cardiac collagen volume fraction (CVF) was examined in picrosirius red stained section using an image analyzing system. Cardiac expression of tissue inhibitor of metalloproteinase 1 (TIMP-1) and transforming growth factor beta 1 (TGF-β1), markers of fibrosis, were determined by Western blot. Compared to eNOS+/+ mice, we found that; 1) myocyte size was significantly increased in eNOS-/- mice; 2) cardiac expression of β-MHC was markedly elevated, while α-SKA levels remained unchanged in eNOS-/- mice; 3) cardiac total and interstitial CVF levels were significantly higher in eNOS-/- mice; and 4) cardiac TIMP-1 levels were significantly greater in eNOS-/- mice, however, cardiac TGF-β1 was not differently expressed between the two groups. The current study revealed that eNOS plays a beneficial role in cardiac remodeling, preventing the heart from development of myocyte hypertrophy and cardiac fibrosis. These findings support our previous report that eNOS may modify the severity of HCM phenotypes.


Geochemical characterization of PM10 emitted by glass factories in Murano, Venice (Italy).
The atmosphere in Venice, like in other European cities, is influenced by complex PM(10) multi-emission sources with a net tendency to exceed the limits fixed by the directive 99/30/EC. This study investigated the composition of an ensemble of similar industrial sources, the Murano Glassmaking Factories (MGFs), and their influence on the Venice air quality, using a modelling approach, statistical analysis and geochemical considerations. Preliminary modelling simulations were conducted to select three sampling sites along the way of preferential transport of pollutants from source between February and April 2003. Subsequently, a sampling campaign was carried out in the same period of simulations. Concentrations of PM(10), eight major elements (Al, Ti, Ca, Mg, Na, K, Fe, Mn), 20 minor and trace elements (Li, V, Cr, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ru, Rh, Cd, Sb, Ba, Ce, Pt, Pb) and four PAHs (BaA, BbF, BkF, BaP) were quantified. The analytical results were statistically processed for exploring the relationships between inorganic elements and organic compounds, and results were interpreted using geochemical considerations. Results show a MGF component of PM(10) characterised by two different fingerprints: the first linked to glass raw material composition and the second mainly related to glass additives. Particularly, Cd, Se, As and Li preserve their ratios in all study area, and are interpreted as principal components of the MGF emissions. Other fingerprints can be traced to urban sources from the Venetian mainland.


The Molecular Mechanism of Domain Swapping of the C-Terminal Domain of the SARS-Coronavirus Main Protease.
In three-dimensional domain swapping, two protein monomers exchange a part of their structures to form an intertwined homodimer, whose subunits resemble the monomer. Several viral proteins domain swap to increase their structural complexity or functional avidity. The main protease (Mpro) of the severe acute respiratory syndrome (SARS) coronavirus proteolyzes viral polyproteins and has been a target for anti-SARS drug design. Domain swapping in the α-helical C-terminal domain of Mpro (MproC) locks Mpro into a hyperactive octameric form that is hypothesized to promote the early stages of viral replication. However, in the absence of a complete molecular understanding of the mechanism of domain swapping, investigations into the biological relevance of this octameric Mpro have stalled. Isolated MproC can exist as a monomer or a domain-swapped dimer. Here, we investigate the mechanism of domain swapping of MproC using coarse-grained structure-based models and molecular dynamics simulations. Our simulations recapitulate several experimental features of MproC folding. Further, we find that a contact between a tryptophan in the MproC domain-swapping hinge and an arginine elsewhere forms early during folding, modulates the folding route, and promotes domain swapping to the native structure. An examination of the sequence and the structure of the tryptophan containing hinge loop shows that it has a propensity to form multiple secondary structures and contacts, indicating that it could be stabilized into either the monomer- or dimer-promoting conformations by mutations or ligand binding. Finally, because all residues in the tryptophan loop are identical in SARS-CoV and SARS-CoV-2, mutations that modulate domain swapping may provide insights into the role of octameric Mpro in the early-stage viral replication of both viruses.


Models of chronic alcohol exposure and dependence.
Alcoholism is a chronic treatment-resistant disorder typically presenting with recurrent/cyclic periods of abusive drinking, withdrawal, abstinence, and relapse. Various strategies that attempt to model these processes in animals have been developed to elucidate the behavioral and neural processes underlying alcoholism. Many of these have involved chronic ethanol exposure and withdrawal with the most widely employed methods involving mice or rats. Prominent features of these methods include alcohol vapor or intragastric forced exposure, cyclic or intermittent periods of alcohol availability with various lengths of forced abstinence, voluntary consumption, the use of genetically alcohol-preferring animals, and inclusion of various pharmacological or environmental challenges to worsen or mitigate symptoms. This chapter emphasizes alcohol exposure and withdrawal and discusses representative metrics used to monitor the consequences of employing these methods. These include but are not limited to intensity and pattern of alcohol exposure, seizure sensitivity during withdrawal, and emotional responding.


The molecular basis for the generation of the human soluble interleukin 2 receptor.
Using an enzyme-linked immunosorbent assay (ELISA) employing two monoclonal antibodies recognizing distinct epitopes on the interleukin 2 receptor (IL2R) alpha chain (Tac molecule), we previously demonstrated that activated lymphocytes release a soluble interleukin 2 receptor molecule (sIL2R) in vitro and in vivo. The sIL2R is biochemically and structurally related to Tac, but its precise origin and functional role remain to be defined. We report here that a single IL2R cDNA is sufficient to direct the synthesis of both cell-associated and soluble released IL2R molecules. Northern analysis of IL2R cDNA transfected L-cell lines revealed the presence of mRNA species unaccounted for by known transcription termination or internal splice sites. Nevertheless, S1 nuclease digestion studies failed to detect alternately spliced mRNA transcripts that specifically lack transmembrane or cytoplasmic domains and which may encode a secreted IL2R molecule. Therefore sIL2R does not appear to be the product of a unique post-transcriptional splicing event. In the absence of any post-translational modifications, sIL2R is most likely generated by enzymatic cleavage and release of cell surface Tac. This proteolytic release of Tac may be but one example of a common cellular mechanism for regulating the membrane expression of cell surface molecules.


Syntheses of chiral N-(protected) tri- and tetrapeptide conjugates.
Cbz-(protected)-tri- and tetrapeptide conjugates with steroids, sugars, terpenes, and heterocycles were prepared using Cbz-(protected)-tri- and tetrapeptidoylbenzotriazoles as active intermediates.


Intersurfactant H-bonds between head groups of n-dodecyl-β-d-maltoside at the air-water interface.
In previous studies we looked at the foam stability of various surfactants with C12 alkyl chains but different head groups and found that stable foams are only generated if the head groups are capable of forming hydrogen bonds with each other. Despite the consistency of the experimental data with the conclusions drawn from it we had no direct proof for our hypothesis that H-bonds are formed between surfactant head groups. To fill this gap, i.e. to demonstrate intersurfactant H-bond formation, we chose the non-ionic sugar surfactant n-dodecyl-β-d-maltoside (β-C12G2) and used molecular dynamics (MD) simulations as well as grazing-incidence X-ray (GIX) scattering and diffraction to study the surfactant-loaded air-water interface. (1) In a densely packed monolayer, close to the critical micelle concentration (cmc), each head group of the sugar surfactant is involved in ∼5 intersurfactant H-bonds with other head groups and in ∼5 H-bonds with water molecules. (2) The number of intersurfactant H-bonds decreases, while the number of surfactant-water H-bonds increases with increasing distance between the head groups (below the cmc). (3) Even at very large distances (well below the cmc) there are still intersurfactant H-bonds, which we ascribe to the formation of clusters at the surface. (4) GIX scattering revealed that a homogeneous surfactant monolayer is formed at full coverage (around the cmc), i.e. cluster formation only happens below the cmc.


Study of Voice Disorders Among School Teachers in Goa.
Teachers are heavy voice users, and they suffer from voice problems more frequently than other occupational voice users. Various studies have demonstrated that teachers speak longer than other professionals and that school teachers in particular, are at risk for voice problems such as vocal fatigue and vocal nodules. The present study is undertaken to study the prevalence of voice disorders in the teachers in different schools at any time and accesses the relationship of different working conditions like class room size, background noise, number of hours taught every day and role of chalk allergy in development of these voice disorders. The study was carried out on 133 school teachers with self reporting of voice problems through detailed questionnaire. A significant number of teachers, more so females had voice problems attributed to various factors. Voice amplification and reduction of background noise along with measures to control allergy are suggested.


The use of unofficial "problem patient" files and interinstitutional information transfer in emergency medicine in Iowa.
The study objectives were to determine the use of "problem patient" files in full-time Emergency Departments (EDs) in the state of Iowa and the frequency of telephone calls made between these departments about drug-seeking patients. The design and setting were a mail survey of 42 EDs in the state of Iowa determined to be staffed full-time by an in-house physician. The participants were the directors of the full time EDs in the state of Iowa. There were no interventions. The measurements and main results are as follows: 38 of 42 sites responded for a response rate of 90%. Of these, 58% keep problem patient lists and consult them an average of 2.6 times per week (range, 0.2 to 7.5). Only 5% have a policy limiting access to the information in these files, and only 14% have any controls over the adding of information. Thirty-seven of the 38 sites reported making and receiving calls to and from other institutions about drug-seeking patients. The mean number of calls made is 23 a year, and the mean number received is 20 per year. Problem patient files are common in EDs in Iowa, as are calls between institutions about drug-seeking patients. These practices compromise patient confidentiality but may be potentially justified in some instances. The lack of controls on access to patient information files and lack of scrutiny of information entered is of concern.


Morphological and biochemical changes associated with apoptosis induced by okadaic acid in human amniotic FL cells.
The marine toxin okadaic acid (OA) is an apoptosis inducer and a tumor promoter. During recent years, extensive studies have demonstrated that OA can induce apoptosis in a wide variety of cell types. In contrast to the relatively longer incubation time or higher treatment concentrations of OA in apoptosis shown previously, relatively lower concentrations (<or=100 nM) and shorter time (4 h) were designed in the current study to observe the toxic effects of OA in human amniotic cells (FL cells). The present study was undertaken to determine the morphological and biochemical changes of FL cells induced by OA. Results indicated that externalization of phosphatidylserine, cytoskeletal disruption, DNA strand breaks and decrease of Bcl-2 protein expression levels as well as increase of PP2A-A subunit protein were all involved in the apoptosis of FL cells induced by OA. This work not only provided further evidence of apoptosis induced by OA but also suggested that PP2A might play a pivotal role in apoptosis induced by protein phosphatases inhibitors.


Secretory production of single-chain antibody (scFv) in Brevibacillus choshinensis using novel fusion partner.
Halophilic β-lactamase (BLA) has been successfully used as a novel fusion partner for soluble expression of aggregation-prone foreign proteins in Escherichia coli cytoplasm (Appl Microbiol Biotechnol 86:649-658, 2010b). This halophilic BLA fusion technology was applied here for secretory expression in Brevibacillus. The "Brevibacillus in vivo cloning" method, recently developed by Higeta Shoyu group, for the construction and transformation of Brevibacillus expression vectors facilitates efficient screening of the production conditions of Brevibacillus expression system. Two single-chain antibodies (scFv), HyHEL-10 single chain scFv (scFvHEL) and anti-fluorescein single chain scFv (scFvFLU), were successfully secreted to culture supernatant as a fusion protein with halophilic BLA. The scFvHEL-His, purified after cleavage of BLA portion with thrombin, was fully active: it formed a stoichiometric complex with the antigen, lysozyme, and inhibited the enzymatic activity. The scFvFLU-His, similarly expressed and purified, stoichiometrically inhibited fluorescence intensity of fluorescein. The molecular mass of scFvHEL-His was determined to be 27,800 Da by light scattering measurements, indicating its monomeric structure in solution.


Structure of simian virus 40 recombinants that contain both host and viral DNA sequences. II. The structure of variant 1103 and its comparison to variant CVPS/1P2 (EcoRI res).
In an effort to characterize sites of recombination between SV40 and monkey DNA, we have determined the primary sequence of a large portion of the SV40 variant, designated 1103. This virus contains DNA sequences derived both from the wild type SV40 genome and from the permissive monkey cell in which the virus was propagated. Further, the monkey sequences included in the defective genome are homologous to both highly repeated monkey DNA (alpha component) and sequences that are infrequently repeated in the monkey genome. The regions of the 1103 genome where DNA sequences were determined include 1) the segments of the variant that surround joints connecting SV40 and monkey sequences, 2) the segment that contains the joint between monkey sequences of high and low reiteration frequency, and 3) the DNA segment of the variant that is homologous to monkey alpha component DNA. Comparison of the data obtained from the sequences analysis of the SV40 variants 1103 and CVP8/1/P2 (EcoRI res) (described in Wakamiya, T., McCutchan, T., Rosenberg, M., and Singer, M. (1979) J. Biol. Chem 254, 3584-3591) reveals certain similarities between the two that may be involved in eukaryotic recombination and defective variant formation.


Spirogermanium--a novel antitumour agent expressing a lack of cross-resistance in vitro with a range of 'standard' antitumour drugs.
We have used a series of drug resistant continuous tumour cell lines of murine, hamster or human origin, to screen in vitro for new drugs effective at overcoming resistance expressed to clinically useful antitumour drugs. We have established that spirogermanium appears to express a unique lack of cross resistance to a wide range of 'standard' antitumour drugs including 5-fluorouracil, cisplatin, methotrexate, vincristine and adriamycin, as judged by its equivalent effectiveness in reducing survival, measured by clonogenic assays, of the parental and drug-resistant sublines following 24 hour in vitro treatments. Spirogermanium was also able to overcome the multidrug resistance exhibited by the colchicine-resistant mutant CHRC5 cells and a subline of vincristine-resistant MCF-7 human breast carcinoma cells, following 1 or 24 hour drug treatments. These preclinical studies suggest that this novel compound may prove a potentially useful agent for inclusion in 'non-cross-resistant drug combinations'.


Practice guidelines for patients with gastrointestinal surgical diseases.
The rauonale for practice gmdehnes has been elucidated It is hoped that these guldehnes will result in changes in chnlcal care Perhaps the origin of these guldehnes may be a cnucal factor m whether they become widely accepted. Our purpose in developing guldehnes for gastrolntesunal surgical disorders is to promulgate reformation that is needed to evaluate patients with diseases that require surgery and to provide mformatlon concerning expected outcomes following specific operattve procedures As pracuce patterns and technology evolve, these guldehnes will need to be changed accordingly.


VGLUT2 expression is up-regulated in neurohypophysial vasopressin neurons of the rat after osmotic stimulation.
A second vesicular glutamate transporter (VGLUT2) is detected in magnocellular neurons in the rat hypothalamus. The present study revealed what phenotype of neurons express VGLUT2 mRNA by the histological method. We found that most vasopressin (VP) neurons and several oxytocin (OT) neurons express VGLUT2 mRNA. VGLUT2 gene expression in VP and OT neurons is enhanced with osmotic challenges. In the neurohypophysis, VGLUT2-staining in OT terminals was reduced with osmotic stimulation. These results indicate that VGLUT2 is principally expressed in VP neurons and also in some OT neurons and that VGLUT2 in VP and OT neurons is involved in osmotic regulation.


COVID-19's effect on the spatial integration of fish markets: Evidence from carp price in China.
While curbing the spread of Coronavirus Disease 2019 (COVID-19), lockdown policies and "stay-at-home" restrictions caused massive supply chain disruptions worldwide. This led to breaks in spatial market integration, which could further lead to market inefficiency and resource misallocation. Taking daily price data from 2016 to 2021, this study investigates COVID-19's effect on the spatial market integration of fish in China using cointegration tests. We find a high degree of spatial market integration for fish in China before the COVID-19 pandemic. Further, our results show that COVID-19's effect on the spatial market integration of fish varies spatially in China. Specifically, COVID-19 reduces the degree of spatial market integration in most provinces, especially those with high infection rates. Meanwhile, the degree of spatial market integration in provinces with low infection rates remains high. Therefore, the government should be regionally specific when formulating market recovery policies.


Asymmetric Stem Cell Division and Germline Immortality.
Germ cells are the only cell type that is capable of transmitting genetic information to the next generation, which has enabled the continuation of multicellular life for the last 1.5 billion years. Surprisingly little is known about the mechanisms supporting the germline's remarkable ability to continue in this eternal cycle, termed germline immortality. Even unicellular organisms age at a cellular level, demonstrating that cellular aging is inevitable. Extensive studies in yeast have established the framework of how asymmetric cell division and gametogenesis may contribute to the resetting of cellular age. This review examines the mechanisms of germline immortality-how germline cells reset the aging of cells-drawing a parallel between yeast and multicellular organisms.


Unveiling the Immunomodulatory Characteristics of Haemonchus contortus Ephrin Domain Containing Protein in the Parasite-Host Interactions.
Ephrin domain containing protein (EPH), a significant excreted and secreted product (ESPs) of Haemonchus contortus, has been identified to have antigenic functions. Over the past years, a new subset of CD4 + T named as T helper 9 cells that secrete interleukin-9 (IL-9) as a signature cytokine is associated with tumor immunity and allergy. Nonetheless, the understanding of immunomodulatory roles of EPH on goat Th9 and other immune cells remains limited. Herein, EPH from H. contortus (HcEPH) was cloned and expressed in pET-28a. Immunofluorescence assay (IFA) was carried-out to localize rHcEPH within H. contortus adult worms and to bind with goat peripheral blood mononuclear cells (PBMCs). Besides, the impact of rHcEPH on signature cytokine IL-9 expression in goat PBMCs was evaluated. Flow cytometry was employed to examine Th9 cells production and cell apoptosis. The results revealed success in the expression and localization of rHcEPH in surface of adult H. contortus gut sections. According to IFA analysis, the rHcEPH protein was capable to react precisely with anti-H. contortus antibodies. Further functional analysis showed that correlation between rHcEPH and host PBMCs significantly enhanced Th9 cell differentiation, IL-9 expression, cell apoptosis efficiency, and cell migration, whereas cell proliferation was suppressed significantly depending on the concentration. Our observations indicated that rHcEPH protein is linked to modulate the host immune cells and could enhance protective immunity by inducing Th9 responses.


Optimal parameters for the destruction of prostate cancer using irreversible electroporation.
Irreversible electroporation is a new tissue ablation technique that consists of applying musecond pulses of direct current to create permanent defects in the cell membrane. Irreversible electroporation spares connective tissue in blood vessels and other tissue structures. When applied properly, it does not produce thermal damage. We determined the irreversible electroporation parameters that would reliably destroy prostate cancer cells. Irreversible electroporation pulses were applied to prostate adenocarcinoma cells in vitro. Three sets of studies were performed to determine the number, length and field strength of irreversible electroporation pulses required to produce complete cancer cell ablation without inducing thermal effects. The outcome of a treatment protocol was simulated. We found the upper and lower limit bounds of pulse length and number in a field range of 2,000 to 250 V/cm. A total of 90 pulses at 250 V/cm for 100 museconds separated by 100 milliseconds could completely ablate prostate cancer cells without inducing thermal damage. Irreversible electroporation represents a new nonthermal ablation modality. This study has produced values for prostate cancer treatment with irreversible electroporation.


Long-term comparative effectiveness of gastric bypass and sleeve gastrectomy on use of antireflux medication: a difference-in-differences analysis.
Gastroesophageal reflux (GERD) is common among patients with obesity who undergo bariatric surgery. Although gastric bypass and sleeve gastrectomy are the most common bariatric operations performed in the United States, their long-term comparative effectiveness on GERD medication use is unknown. To compare the long-term effectiveness of gastric bypass and sleeve gastrectomy on use of antireflux medication. National cohort undergoing inpatient bariatric surgery. This is a retrospective study of Medicare beneficiaries undergoing gastric bypass and sleeve gastrectomy between January 1, 2012, and December 31, 2017. A difference-in-differences analysis was conducted to evaluate the differential change in antireflux medication use between groups before and after surgery. A total of 16,640 patients underwent gastric bypass, and 26,724 patients underwent sleeve gastrectomy. Before surgery, GERD medication use was higher among patients who underwent gastric bypass (62.4%; 95% confidence interval [CI]: 62.0%-63.7%) compared with patients who underwent sleeve gastrectomy (60.1%; 95% CI: 59.3%-60.9%). Five years after surgery, GERD medication use was lower in patients who underwent gastric bypass (47.8%; 95% CI: 46.3%-49.3%) compared with patients who underwent sleeve gastrectomy (53.7%; 95% CI: 50.5%-56.9%). The differential decrease from baseline GERD medication use was greater for patients who underwent gastric bypass at 2 years (-4.1 percentage points [pp]; 95% CI: -1.7 to -6.5 pp), 3 years (-4.3 pp; 95% CI: -1.6 to -7.0 pp), 4 years (-6.9 pp; 95% CI: -4.1 to -9.6 pp), and 5 years (-8.3 pp; 95% CI: -3.7 to 12.8 pp) after surgery. Though use of antireflux medication decreased following both procedures, gastric bypass was associated with a greater reduction in antireflux medication use 5 years after surgery compared with sleeve gastrectomy. Understanding the long-term comparative effectiveness of these common bariatric operations may better inform treatment decisions among patients and surgeons.


Telemedicine and the current opportunities for the management of oncological patients in Peru in the context of COVID-19 pandemic.
COVID-19 pandemic is the more challenging public health emergency of the century, producing the collapse of health systems and unprecedented levels of morbidity and mortality around the world, especially in low resource settings. Patients with chronic diseases are the most affected, not only due to the high susceptibility to SARS-CoV-2 infection but also due to the decrease in opportunities for timely care. In this dark landscape, telemedicine, before limited to very specific scenarios, has become one of our main tools to manage cancer patients, particularly in Latin America where COVID-19 has had a strong impact on the public health. Telemedicine can provide rapid access to specialized cancer care in a scenario complicated, reducing the exposure of patients and healthcare personnel to the SARS-CoV-2. In this review, we would like to share our experience and our workflow using telemedicine at Oncosalud-AUNA, a private clinic in Peru.


Identification of novel β3-adrenoceptor agonists using energetic analysis, structure based pharmacophores and virtual screening.
β3 Adrenergic receptor (β3-AR), is a potential therapeutic target for the treatment of type II diabetes and obesity. We report the identification of novel compounds as β3-AR agonists by integrating different approaches of energetic analysis, structure based pharmacophore designing and virtual screening. In a step wise filtering protocol, structure based virtual screening of 2, 33, 450 compounds was done. These molecules were docked into the active site of the receptor utilizing three levels of accuracy; ligands passing the HTVS (high throughput virtual screening) step were subsequently analyzed in Glide SP (Standard Precision) and finally in Glide XP (Extra Precision) to estimate the receptor ligand binding affinities. In the second step a total of 300 pharmacophore hypotheses were generated from a set of known and diverse β3-AR agonists. The best hypothesis showed six features: three hydrogen bond acceptors, one positively charged group, and two aromatic rings. To cross validate, pharmacophore filtering was done on the set of shortlisted compounds from structure based VS (virtual screening). The different screening techniques employed were validated using enrichment factor calculations. The energetic based Pharmacophore performed fairly well at distinguishing active from the inactive compounds and yielded a greater diversity of active molecules whereas the number of actives retrieved in the case of structure based screening was the highest.


Effect of tensile force on expression of PTHrP and thickness of hypertrophic zone in organ-cultured mouse spheno-occipital synchondroses.
Responses of spheno-occipital synchondroses to direct tensile stress have not been identified before. This study was, therefore designed to evaluate expression of PTHrP, and thickness of hypertrophic zone in spheno-occipital synchondroses in response to such stress, using mouse in vitro model. Spheno-occipital synchondroses together with adjacent structures were excised from fifty-five 2-day-old mice that were randomly assigned to 6 control and 5 experimental groups for 5 experimental periods (n=5). In the experimental groups, tensile force of 0.2g was applied across the synchondroses, using helical springs. In 5 control groups, the springs were made inactive. Both groups were then cultured for 6, 24, 48, 72 h and 7 days. Another control group was cultured without any springs for 7 days to compare with natural growth of the synchondroses from a group of five 9-day-old mice. Alcian blue-PAS staining was used to study growth of the synchondroses; immunohistochemical staining to identify PTHrP and type X collagen expression. The area of PTHrP expression and thickness of hypertrophic zone, demarcated by type X collagen expression, were measured. Quantitative analysis showed that PTHrP expression increased significantly at hour 24 of the force application in the experimental group (p<0.05), then reduced from hour 24 to 72 with a significant drop from hour 24 to 48 (p<0.01); and the thickness of hypertrophic zone significantly increased at hour 48 (p<0.01). Our findings suggested that the growth of spheno-occipital synchondroses could be modified by tensile stress; and a light continuous force could enhance its growth, as evidenced by an increase in PTHrP expression and thickness of hypertrophic zone.


Regulation of diacylglycerol production and protein kinase C stimulation during sperm- and PLCzeta-mediated mouse egg activation.
At fertilization in mammalian eggs, the sperm induces a series of Ca(2+) oscillations via the production of inositol 1,4,5-trisphosphate. Increased inositol 1,4,5-trisphosphate production appears to be triggered by a sperm-derived PLCzeta (phospholipase C-zeta) that enters the egg after gamete fusion. The specific phosphatidylinositol 4,5-bisphosphate hydrolytic activity of PLCzeta implies that DAG (diacylglycerol) production, and hence PKC (protein kinase C) stimulation, also occurs during mammalian egg fertilization. Fertilization-mediated increase in PKC activity has been demonstrated; however, its precise role is unclear. We investigated PLCzeta- and fertilization-mediated generation of DAG in mouse eggs by monitoring plasma-membrane translocation of a fluorescent DAG-specific reporter. Consistent plasma-membrane DAG formation at fertilization, or after injection of physiological concentrations of PLCzeta, was barely detectable. However, when PLCzeta is overexpressed in eggs, significant plasma-membrane DAG production occurs in concert with a series of unexpected secondary high-frequency Ca(2+) oscillations. We show that these secondary Ca(2+) oscillations can be mimicked in a variety of situations by the stimulation of PKC and that they can be prevented by PKC inhibition. The way PKC leads to secondary Ca(2+) oscillations appears to involve Ca(2+) influx and the loading of thapsigargin-sensitive Ca(2+) stores. Our results suggest that overproduction of DAG in PLCzeta-injected eggs can lead to PKC-mediated Ca(2+) influx and subsequent overloading of Ca(2+) stores. These results suggest that DAG generation in the plasma membrane of fertilizing mouse eggs is minimized since it can perturb egg Ca(2+) homoeostasis via excessive Ca(2+) influx.


A chromosome-level genome assembly of tomato pinworm, Tuta absoluta.
The tomato pinworm, Tuta absoluta, or Phthorimaea absouta, is native to South America, but quickly spread to other regions of world, including Europe, Africa, and Asia, devastating to global tomato production. However, a lack of high-quality genome resources makes it difficult to understand its high invasiveness and ecological adaptation. Here, we sequenced the genome of the tomato pinworm using Nanopore platforms, yielding a genome assembly of 564.5 Mb with contig N50 of 3.33 Mb. BUSCO analysis demonstrated that this genome assembly has a high-level completeness of 98.0% gene coverage. In total, 310 Mb are repeating sequences accounting for 54.8% of genome assembly, and 21,979 protein-coding genes are annotated. Next, we used the Hi-C technique to anchor 295 contigs to 29 chromosomes, yielding a chromosome-level genome assembly with a scaffold N50 of 20.7 Mb. In sum, the high-quality genome assembly of the tomato pinworm is a useful gene resource that contributes to a better understanding of the biological characteristics of its invasiveness and will help in developing an efficient control policy.


The effect of Agonoscena targionii (Licht.) (Homoptera: Psylloidea) on seed production by Pistacia terebinthus L.
This study investigated the potential negative effect that psyllids may have on the production of viable seeds in Pistacia terebinthus, a wild plant species with a circummediterranean distribution. Manipulative experiments, involving the application of insecticide, showed that the reduction in fruit maturation is rather low, not exceeding 10%. This is mainly due to other factors that determine seed inviability. Such factors include those intrinsic to the plant, such as parthenocarpy and seed abortion, and extrinsic to it, such as seed predation by chalcidoid wasps. There is much variation in the frequency of psyllids per plant; such variation appears uncorrelated with variables decribing plant size, fecundity, and distance to fruiting conspecifics. No consistency in the incidence of psyllids on a given individual plant was found between years.


The perfect timing for multimodal integration is not the same in all L5 neurons.
In this issue of Neuron, Rindner et al. (2022) demonstrate that subclasses of layer 5 pyramidal neurons in the parietal cortex integrate inputs from frontal and sensory areas supralinearly and with distinct temporal dynamics.


Production of insulin-like growth factor-binding proteins during the development of hepatic fibrosis due to chronic hepatitis C.
Insulin-like growth factor 1 is modulated by the insulin-like growth factor-binding proteins (IGFBPs) that are synthesized in the liver. The aim of the present study was to evaluate the concentrations of IGFBPs 1-7 in patients with chronic hepatitis C and study their association with fibrosis stage. A prospective, cross-sectional study was conducted that included patients with chronic hepatitis C. The stages of fibrosis were determined through FibroTest and FibroScan and the patients were compared with a control group. Serum levels of IGFBPs 1-7 were quantified through multiple suspension arrays. The Kruskal-Wallis test, Mann-Whitney U test, Spearman's correlation, and ROC curves were used for the statistical analysis. Upon comparing the patients and controls, the highest concentrations were found in IGFBPs 1, 2, 4, and 7 (p=0.02, p=0.002, p=0.008, and p<0.001, respectively). IGFBP-3 levels had a tendency to be lower in the patients (p=0.066), whereas values were similar between patients and controls for IGFBP-5 and 6 (p=0.786 and p=0.244, respectively). Of the seven IGFBPs, IGFBP-3 concentrations were the highest. There were significant differences between fibrosis stages for IGFBP-5 and IGFBP-7. IGFBPs play a relevant role in the fibrotic process in liver damage. IGFBP-7, in particular, differentiates fibrosis stages, making it a potential serum biomarker.


Expression of maedi-visna virus major core protein, p25: development of a sensitive p25 antigen detection assay.
The gene for the major core protein, p25, of maedi-visna virus (MVV) was cloned using a PCR (polymerase chain reaction) strategy employing primers designed for the insertion of the gene directly into yeast Ty-VLP expression vectors. In this system p25 is expressed as a fusion protein which self-assembles into virus-like particles (VLPs) due to interaction of the Ty A fusion partner. High levels (50-60 mg/l) of p25 fusion protein were produced, and p25 was recovered in soluble and highly pure form following cleavage from the Ty particle by Factor Xa protease digestion. The p25 protein produced in yeast is antigenically authentic, as defined by its reactivity with p25-specific antisera and its ability to elicit antibodies reactive with native viral p25 protein; although the cleaved, soluble form of p25 was found to be considerably more antigenic than the hybrid Ty-p25 VLP. Using this reagent anti-p25 monoclonal and polyclonal antibodies were generated. These sera and the p25 protein have been used to develop a sensitive MVV p25 detection assay. These reagents and assays will facilitate further studies of viral replication and immune response to the virus.


Sleep Quality and Poor Sleep-related Factors Among Healthcare Workers During the COVID-19 Pandemic in Vietnam.
The coronavirus disease 2019 (COVID-19) pandemic has increased the workload of healthcare workers (HCWs), impacting their health. This study aimed to assess sleep quality using the Pittsburgh Sleep Quality Index (PSQI) and identify factors associated with poor sleep among HCWs in Vietnam during the COVID-19 pandemic. In this cross-sectional study, 1000 frontline HCWs were recruited from various healthcare facilities in Vietnam between October 2021 and November 2021. Data were collected using a 3-part self-administered questionnaire, which covered demographics, sleep quality, and factors related to poor sleep. Poor sleep quality was defined as a total PSQI score of 5 or higher. Participants' mean age was 33.20±6.81 years (range, 20.0-61.0), and 63.0% were women. The median work experience was 8.54±6.30 years. Approximately 6.3% had chronic comorbidities, such as hypertension and diabetes mellitus. About 59.5% were directly responsible for patient care and treatment, while 7.1% worked in tracing and sampling. A total of 73.8% reported poor sleep quality. Multivariate logistic regression revealed significant associations between poor sleep quality and the presence of chronic comorbidities (odds ratio [OR], 2.34; 95% confidence interval [CI], 1.17 to 5.24), being a frontline HCW directly involved in patient care and treatment (OR, 1.59; 95% CI, 1.16 to 2.16), increased working hours (OR, 1.84; 95% CI,1.37 to 2.48), and a higher frequency of encountering critically ill and dying patients (OR, 1.42; 95% CI, 1.03 to 1.95). The high prevalence of poor sleep among HCWs in Vietnam during the COVID-19 pandemic was similar to that in other countries. Working conditions should be adjusted to improve sleep quality among this population.


Oxygen dependence of salbutamol elimination by the isolated perfused rat liver.
Although impairment of drug metabolism by severe hypoxia is well documented in perfused liver preparations, the degree of hypoxia required to produce inhibition of drug elimination pathways in the intact liver has not been defined. In this study, in the isolated perfused rat liver, we examined the relationship between the rate of hepatic oxygen supply and the elimination rate of the drug salbutamol, which in the rat liver is eliminated largely by glucuronidation. Livers (N = 15) from male Sprague-Dawley rats were perfused in a non-recycling design with 10% human red cells in a Krebs-Henseleit electrolyte solution. Salbutamol elimination was examined during normal oxygenation (perfusate equilibrated with 100% O2; mean O2 delivery 3.21 mumol/min/g liver), at a given lower rate of oxygen delivery (achieved by producing different mixtures of N2 with O2 in the perfusate oxygenator) and after reoxygenation. In these experiments, hepatic clearance of salbutamol (perfusate concentration 50 ng/ml) was essentially independent of oxygen delivery above a rate of 2.0 mumol/min/g liver; below this level, clearance fell linearly as O2 supply was reduced. In all livers, reoxygenation restored drug elimination to control levels. In further experiments using a recycling design (N = 22), the effect of hypoxia on salbutamol elimination was found to be very similar. In recycling normoxic experiments (N = 3), the glucuronide metabolite was detected in perfusate and bile, but no sulphate metabolite was detected. While previous studies indicate that elimination of some oxidatively metabolised substrates is very sensitive to reductions in hepatic oxygenation, the present study shows that, in the isolated liver, large reductions in hepatic oxygen supply were required to produce significant impairment of the glucuronidation-dependent elimination of salbutamol.


Long-term outcomes of mini-sternotomy versus conventional sternotomy for aortic valve replacement: a randomized controlled trial.
Aortic valve replacement (AVR) for severe symptomatic aortic stenosis is one of the most common cardiac surgical procedures with excellent long-term outcomes. Multiple previous studies have compared short-term outcomes of AVR with mini-sternotomy versus AVR with conventional sternotomy. We have previously reported the results of the randomized MAVRIC trial, which aimed to evaluate early postoperative morbidity among patients undergoing mini-sternotomy and conventional sternotomy AVR. We now report the long-term all-cause mortality, reoperation, MACE outcomes and echocardiographic data from this trial. The prospective, randomized, single-centre, single-blind MAVRIC (manubrium-limited mini-sternotomy versus conventional sternotomy for aortic valve replacement) trial compared manubrium-limited mini-sternotomy and conventional median sternotomy for the treatment of patients with severe aortic stenosis. The previously reported primary outcome was the proportion of patients receiving red cell transfusion postoperatively and within 7 days of the index procedure. Currently reported exploratory analyses of a combined long-term all-cause mortality and reoperation were compared between groups via the log-rank test. Sensitivity analyses reviewed individual components of the combined end point. The primary analysis and long-term exploratory analyses were based on an intention-to-treat principle. Between March 2014 and June 2016, 270 patients were enrolled and randomized in a 1:1 fashion to undergo mini-sternotomy AVR (n = 135) or conventional median sternotomy AVR (n = 135). At the median follow-up of 6.1 years, the composite outcome of all-cause mortality and reoperation occurred in 18.5% (25/135) of patients in the conventional sternotomy group and in 17% (23/135) of patients in the mini-sternotomy group. The incidence of chronic kidney disease, cerebrovascular accident and myocardial infarction was not significantly different between 2 groups. Follow-up echocardiographic data suggested no difference in peak and mean gradients or incidence of aortic regurgitation between 2 approaches. This exploratory long-term analysis demonstrated that, in patients with severe aortic stenosis undergoing isolated AVR, there was no significant difference between manubrium-limited mini-sternotomy and conventional sternotomy with respect to all-cause mortality, rate of reoperation, MACE events and echocardiographic data at the median of 6.1-year follow-up.


Social transformation and social isolation of older adults: Digital technologies, nursing, healthcare.
The incidence of social isolation among older adults is on the rise in today's health care climate. Consequently, preventing or ameliorating social isolation through technology in this age group is now being discussed as a significant social and health issue. The purpose of the opinion paper is to clarify social transformation through technology and shed light on a new reality for older adults in situations of social isolation. Our goal is to persuade the reader that our position on this topic is a valid one. We support our claims with practice-based evidence and published research studies. To do so, we checked the most recent literature, most of which came from the last decade. Our literature survey focused primarily on what is known about technology and how technology can affect social transformation and perceptions of social isolation. Two dominant transformative realities became the focal points: the precarious implications of loneliness for older adults and the emerging reality of social change through digital technology central to eHealth and mHealth. To benefit from new technologies and reduce the detrimental effects of social isolation, we must engage older adults in a meaningful way and adapt the system of smart devices to reflect the specific physiological and psychological characteristics of the ageing population. Older adults need to comprehend the meanings of their social experiences to preserve their active lifestyle. Human interactions may be desirable, but technological dominance may also minimize the adverse effects of social isolation.


Leukemic predisposition of pSca-1/Cb2 transgenic mice.
The gene encoding the peripheral cannabinoid receptor Cb2 is located in the common virus integration site Evi11 and is associated with hematopoietic malignancies in mice. To determine the effect of Cb2 overexpression on hematopoietic development in vivo, Cb2 transgenic mice were generated. A Cb2 expression vector was constructed containing a Cb2 cDNA fragment cloned into the 14kb Sca-1 (Ly-6E.1) gene. Two transgenic lines in which Cb2 expression is controlled by the Sca-1 promoter were generated, and the effect on hematopoietic development was studied. Expression of Cb2 mRNA or protein was studied by RNase protection analysis and ligand binding assays, respectively. Leukemic predisposition was investigated by injecting newborn transgenic as well as control animals with Cas-Br-M murine leukemia virus (Cas-Br-M MuLV). Although increased expression of the Cb2 gene was observed in hematopoietic tissues, follow-up of more than 1 year did not reveal any hematologic defect. Interestingly, infection of newborn pSca-1/Cb2 transgenic mice with Cas-Br-M MuLV revealed that significantly more transgenic mice developed leukemia than virus-treated control littermates. Because these studies provide evidence for the cooperative potential of Cb2 in leukemia progression, we wished to identify genes that may collaborate with Cb2 in leukemic transformation. Our study suggests that Evi1, another common target for proviral integration in mouse leukemias, may be overexpressed in virus-induced leukemias in pSca-1/Cb2 transgenic mice. The data indicate that hematopoietic precursor cells that express high levels of Cb2 possess increased susceptibility for leukemia development and that Cb2 and Evi1 might collaborate in leukemogenesis.


Heart Failure: Priorities for Transition to Home.
Nurses play a key role in promoting successful transitions of patients with heart failure (HF) from the hospital to the ambulatory setting. Engaging patients and caregivers in discharge teaching early in the hospitalization can enhance their understanding of HF as a clinical syndrome and identify precipitants of decompensation. Effective transitional care interventions for patient with HF include a phone call within 48 to 72 hours and a follow-up appointment within 7 days. Early symptom identification and treatment are key aspects of HF care to improve quality of life and minimize risk of hospitalization.


Clonal Hematopoiesis Has Prognostic Value in Dilated Cardiomyopathy Independent of Age and Clone Size.
Clonal hematopoiesis (CH) gives rise to mutated leukocyte clones that induce cardiovascular inflammation and thereby impact the disease course in atherosclerosis and ischemic heart failure. CH of indeterminate potential refers to a variant allele frequency (VAF; a marker for clone size) in blood of ≥2%. The impact of CH clones-including small clone sizes (VAF <0.5%)-in nonischemic dilated cardiomyopathy (DCM) remains largely undetermined. The authors sought to establish the prognostic impact of CH in DCM including small clones. CH is determined using an ultrasensitive single-molecule molecular inversion probe technique that allows detection of clones down to a VAF of 0.01%. Cardiac death and all-cause mortality were analyzed using receiver-operating characteristic curve-optimized VAF cutoff values. A total of 520 DCM patients have been included. One hundred and nine patients (21%) had CH driver mutations, of which 45 had a VAF of ≥2% and 31 <0.5%. The median follow-up duration was 6.5 years [IQR: 4.7-9.7 years]. DCM patients with CH have a higher risk of cardiac death (HR: 2.33 using a VAF cutoff of 0.36%, 95% CI: 1.24-4.40) and all-cause mortality (HR: 1.72 using a VAF cutoff of 0.06%, 95% CI: 1.10-2.69), independent of age, sex, left ventricular ejection fraction, and New York Heart Association classification. CH predicts cardiac death and all-cause mortality in DCM patients with optimal thresholds for clone size of 0.36% and 0.06%, respectively. Therefore, CH is prognostically relevant, independent of clone size in patients with DCM.


Disruptions in intracellular membrane trafficking and structure preclude the glucocorticoid-dependent maturation of mouse mammary tumor virus proteins in rat hepatoma cells.
We have previously shown that glucocorticoids regulate the trafficking and processing of mouse mammary tumor virus (MMTV) proteins in viral-infected M1.54 rat hepatoma cells. To examine the role of intracellular membrane integrity on MMTV protein maturation, brefeldin A (BFA) was utilized to disrupt membrane flow between the endoplasmic reticulum and Golgi. Immunoprecipitation and immunofluorescence microscopy revealed that in the presence of dexamethasone, BFA inhibited the proteolytic processing, cell surface delivery, and externalization of MMTV glycoproteins. Glycosidase digestion and inhibitors of protein glycosylation confirmed that the observed differences in apparent sizes of MMTV glycoprotein products are due to BFA-induced changes in oligosaccharide processing. BFA treatment inhibited the proteolytic processing of the MMTV phosphoprotein precursor, which normally associates with the cytoplasmic face of intracellular membranes. Similarities in salt extraction efficiency revealed that BFA did not affect the membrane affinity of the uncleaved phosphorylated precursor. In a complementary approach, proteolytic processing of the phosphorylated polyprotein did not occur in glucocorticoid-treated HTC cells transfected with a mutant MMTV provirus encoding a normal phosphorylated precursor, but which express a truncated MMTV glycoprotein missing its transmembrane domain and cytoplasmic tail. These results suggest that the MMTV glycoproteins and phosphoproteins may interact at a late step in the transport pathway in a manner required for their mutual processing in response to glucocorticoids and establishes the importance of functional interactions with intracellular membranes for maturation of the cytoplasmic MMTV phosphoproteins.


Association Between Emergency Physician Length of Stay Rankings and Patient Characteristics.
Emergency physicians are commonly compared by their patients' length of stay (LOS). We test the hypothesis that LOS is associated with patient characteristics and that accounting for these features impacts physician LOS rankings. This was a retrospective observational study of all encounters at an emergency department in 2010 to 2015. We compared the characteristics of patients seen by physicians in different quartiles of LOS. Primary outcome was variation in patient characteristics at time of physician assignment (age, sex, comorbidities, Emergency Severity Index [ESI], and chief complaint) across LOS quartiles. We also quantified the change in LOS rankings after accounting for difference in characteristics of patients seen by different physicians. A total of 264,776 encounters seen by 62 attending physicians met inclusion criteria. Physicians in the longest LOS quartile saw patients who were older (age = 49.1 vs 48.6 years, difference = +0.5 years, 95% confidence interval [CI] = 0.3 to 0.7) with more comorbidities (Gagne score = 1.3 vs. 0.9, difference = +0.4, 95% CI = 0.4 to 0.4) and higher acuity (ESI = 2.8 vs. 2.9, difference = -0.1, 95% CI = 0.1 to 0.1) than physicians in the shortest LOS quartile. The odds ratio (OR) of physicians in the longest LOS quartile seeing patients over age 50 compared to the shortest LOS quartile was 1.1 (95% CI = 1.0 to 1.1); the OR of physicians in the longest LOS quartile seeing patients with ESI of 1 or 2 was also 1.1 (95% CI = 1.0 to 1.1). Accounting for variation in patient characteristics seen by different physicians resulted in substantial reordering of physician LOS rankings: 62.9% (39/62) of physicians reclassified into a different quartile with mean absolute percentile change of 25.8 (95% CI = 20.3 to 31.3). A total of 62.5% (10/16) of physicians in the shortest LOS quartile and 56.3% (9/16) in the longest LOS quartile moved into a different quartile after accounting for variation in patient characteristics. Length of stay was significantly associated with patient characteristics, and accounting for variation in patient characteristics resulted in substantial reordering of relative physician rankings by LOS. Comparisons of emergency physicians by LOS that do not account for patient characteristics should be reconsidered.


Trends in birth rates for teenagers in Queensland, 1988 to 1997: an analysis by economic disadvantage and geographic remoteness.
To investigate trends in the birth rate for teenagers in Queensland, stratified by geographic remoteness and economic disadvantage. This was an analysis of routine data for the period 1988 to 1997. The number of births were obtained from the Queensland Perinatal Data Collection. Population data (the denominators for the rates) were obtained from the Australian Bureau of Statistics. Economic disadvantage was based on place of usual residence of the mother. Because of differences in physical, social and psychological development, the data were analysed in three age groups: 13 to 14 years, 15 to 17 years and 18 to 19 years. Birth rates to teenagers who live in disadvantaged areas were 2 to 4 times higher than the rates for all of Queensland and 10 to 20 times higher than the rates in affluent areas. The trend analysis showed that the rates are decreasing in urban-affluent areas (about 2.5% per year), while they have remained stable, but extremely high in disadvantaged-remote areas, and are increasing (about 5% per year) in disadvantaged-urban areas. The relatively low and stable rates for all of Queensland have hidden marked variations in the trends for areas defined according to economic disadvantage and remoteness. The rates were especially high and showed no improvement over time in remote disadvantaged areas, which have a large indigenous population. In some areas of Queensland births to teenagers is a pressing problem, especially because it can perpetuate a cycle of limited educational opportunities, social isolation and reliance on welfare.


Directed mutagenesis and plasmid-based complementation in the methanogenic archaeon Methanosarcina acetivorans C2A demonstrated by genetic analysis of proline biosynthesis.
We report here the first use of directed mutagenesis in Methanosarcina acetivorans C2A. The method employs homologous recombination-mediated gene replacement and was used to construct a variety of proline auxotrophs with mutations in the proABC locus. Each mutation was also complemented in trans with autonomously replicating Methanosarcina-Escherichia plasmid shuttle vectors.


Flavonoid glycosides and an anthraquinone from Rumex chalepensis.
Besides rutin, quercetin 3-rhamnoside and kaempferol 3-rhamnosyl(1-->6) galactoside, a new flavonol glycoside, quercetin 3-glucosyl(1-->4)galactoside, and 1,6,8-trihydroxy-3-methyl anthraquinone (emodin) have been characterized from leaves of Rumex chalepensis. The structures were established on the basis of Rf values, acid hydrolysis to aglycone and sugar and UV, EI and FAB-mass spectra, 1H NMR, 13C DEPT NMR, NOE difference measurement, 1H-H COSY and 1H-13C COSY spectral data.


HISTORICAL BIOGEOGRAPHY IN THE INDO-PACIFIC: A CLADISTIC APPROACH.
Abstract- The history of biogeographic theories about the Indo-Pacific is reviewed. The methods of cladistic biogeography are introduced and 10 monophyletic groups of Miridae (Insecta: Heteroptera) are used to apply these methods in the Indo-Pacific. The results are summarized and then combined with those of Duffels on cicadas. The results of most previous schemes are rejected on the basis of new results because the prior studies applied methods which failed to correctly interrelate areas of endemism within the Indo-Pacific and between this and other regions. The composite biotic nature of New Guinea is stressed. "it is the presence in and not the absence from land areas of organisms that calls for reasoned explanations" (Evans, 1959).


Water relations of Paecilomyces variotii, Eurotium amstelodami, Aspergillus candidus and Aspergillus sydowii, xerophilic fungi isolated from Indonesian dried fish.
The water relations of four xerotolerant fungi, Paecilomyces variotii, Eurotium amstelodami, Aspergillus candidus and Aspergillus sydowii, isolated from dried salt fish, were examined at 25 degrees C, on media in which water activity (aW) was controlled by NaCl or a glucose/fructose mixture. All fungi were less tolerant of NaCl than glucose/fructose at low aW. P. variotii grew 2 to 3 times faster on glucose/fructose media than on NaCl. The minimum aW permitting germination varied from 0.753 for E. amstelodami and, 0.776 for A. candidus and A. sydowii to 0.793 for P. variotii. At low aW germination was not always followed by growth. In most cases the minimum for growth was 0.02 aW units above that for germination.


Stand Your Ground: Policy and Trends in Firearm-Related Justifiable Homicide and Homicide in the US.
In recent years, several states have enacted Stand Your Ground (SYG) legislation intended to deter crime. To date, the impact of these laws on justifiable homicide (JH) and homicide (HOM) is unknown. This study was designed to compare JH and HOM rates (JHR, HR) before and after enactment of SYG laws and in states with and without (NoSYG) such legislation. This was a retrospective analysis of justifiable homicide and homicide using 3 United States datasets from 2000 to 2017. Descriptive statistics and univariate linear regressions were used to evaluate the association of SYG laws and JH and HOM in the United States. There were 4,594 incidents of JH and 214,630 incidents of HOM in the United States between 2000 and 2017. Stand Your Ground states accounted for most JH and HOM incidents 3,184 (69%) and 116,511 (54%), respectively. Stand Your Ground states had higher JH and HOM rates than NoSYG states (0.126 vs 0.047 and 4.663 vs 3.301 per 100,000 population, respectively; p < 0.001). Comparing periods pre- (2000 to 2004) and post-SYG (2005 to 2017) law enactment, SYG vs NoSYG states had increased JH rates of 54.9% vs 20.4%, respectively (p < 0.001); HOM rates increased 10.8% in SYG states and decreased 2.3% in NoSYG states (p < 0.001). Stand Your Ground states had significantly higher JH and HOM rates and significantly greater increases in JH and HOM rates pre- vs post-SYG law enactment compared with NoSYG states. These results raise questions about the potential adverse consequences of policies such as the SYG law in the US.


The study of food addiction using animal models of binge eating.
This review summarizes evidence of "food addiction" using animal models of binge eating. In our model of sucrose bingeing, behavioral components of addiction are demonstrated and related to neurochemical changes that also occur with addictive drugs. Evidence supports the hypothesis that rats can become dependent and "addicted" to sucrose. Results obtained when animals binge on other palatable foods, including a fat-rich food, are described and suggest that increased body weight can occur. However, the characterization of an addiction-like behavioral profile in animals with binge access to fat requires further exploration in order to dissociate the effect of increased body weight from the diet or schedule of feeding.


Opposite effects of neonatal hypoxia on acute amphetamine-induced hyperlocomotion in adult and adolescent mice.
We investigated whether the effect of neonatal hypoxia on amphetamine-induced hyperlocomotion can reproduce the ontogenic (age of onset) properties of schizophrenia. Neonatal hypoxia enhanced amphetamine-induced hyperlocomotion in adult mice and decreased it in adolescent mice. These findings provide ontogenic validity for this very simple animal model of schizophrenia.


Sister chromatid exchanges in human lymphocytes exposed to single cytotoxic drugs in vivo or in vitro.
Venous blood was taken from apparently healthy volunteers and patients with cancer, the latter both before and at intervals after treatment with single cytotoxic drugs. Cells from untreated individuals were exposed to a range of concentrations of drugs in culture medium. Chlorambucil, treosulfan and cyclophosphamide (activated by hepatic microsomes) significantly increased the numbers of SCEs, both in vitro and in the lymphocytes of patients. While methotrexate and 5-fluorouracil had no effect, bleomycin slightly increased the number of SCEs, but only in vitro. Although the in vitro dose-effect relationship indicated which drugs would increase the frequency of SCE in vivo, the magnitude of the response tended to be overestimated. When patients were treated with drugs the frequency of SCE increased, then declined with time. Though this complicates the quantitative relationship between dose and damage, SCEs may be useful to monitor the effects of alkylating agents on normal tissues.


Mimics and rare presentations of pediatric demyelination.
This article reviews the features that should prompt consideration of diseases that mimic acquired demyelinating syndromes and multiple sclerosis using vignettes to highlight unusual clinical and radiologic features. Cases of transverse myelitis, spinal infarction, acute disseminated encephalomyelitis, fever-induced refractory epileptic encephalopathy in school-aged children, small-vessel vasculitis, Griscelli syndrome type 2, cysticercosis, vitamin B12 deficiency, and chronic relapsing inflammatory optic neuropathy are presented.


Bilateral abducens nerve palsy due to septic thrombophlebitis.
To report a case of septic thrombophlebitis producing bilateral abducens nerve palsy. A 65 year-old woman with recent sinus surgery experienced the onset of horizontal diplopia during treatment for bacteremia. Computer tomography of head and a neck ultrasonography showed right internal jugular vein occlusion. Ophthalmology examination was consistent with bilateral abducens nerve palsy. She was treated with systemic antibiotics and antiplatelet therapy with resolution of the internal jugular vein occlusion. The diplopia improved over a six-months. Our patient had Lemierre syndrome with an unusual presentation. The patient was treated for septic thrombophlebitis with a resolution of her ocular symptoms.


Emergency nurse perceptions of individual and facility emergency preparedness.
Disasters of any variety can occur at any given moment. Emergency departments are at the forefront of the response system, serving as the gateway to the most appropriate care of victims. The intent of the survey described in this article was to assess nurses' perception of their role in a disaster and their perceived susceptibility to a disaster. In addition, basic knowledge and role preparation was reviewed. A descriptive survey using survey methodology was utilized. The 56-question survey, including 16 demographic questions, was developed for the purpose of this study. The results reflect that many emergency nurses have not taken basic actions to prepare themselves for a disaster, either personally or professionally. This article highlights the importance of disaster education geared to the needs of the emergency nurse.


A particulate basis for a lattice-gas model of amphiphilic fluids.
We show that the flux-field expansion derived by Boghosian & Coveney for the Rothman-Keller immiscible fluid model can be derived in a simpler and more general way in terms of the completely symmetric tensor kernels introduced by those authors. Using this generalized flux-field expansion we show that the more complex amphiphilic model of Boghosian, Coveney & Emerton can also be derived from an underlying model of particle interactions. The consequences of this derivation are discussed in the context of previous equilibrium Ising-like lattice models and other non-equilibrium mesoscale models.


Constitutive activation of Stat1 and Stat3 in primary erythroleukemia cells.
Signal transducers and activators of transcription (Stat) proteins play important roles in the regulation of hematopoiesis as downstream molecules of cytokine signal transduction. Previously, we demonstrated that Stat1 and Stat3 are activated by erythropoietin (EPO) in a human EPO-dependent erythroleukemia cell line UT-7/EPO. We report here that Stat1 and Stat3 are constitutively activated in freshly isolated erythroleukemia cells. In addition, EPO promoted cell growth of these cells, accompanied by enhanced activities of Stat1 and Stat3. Furthermore, mutation in the Statl/Stat3-binding sites of the c-myc gene promoter clearly blocked its promoter activity in EPO-stimulated primary erythroleukemia cells. Thus, Stat1 and Stat3 may support cell growth in part via c-myc gene activation in primary erythroleukemia cells.


COVID-19 and Subjective Well-Being in Urban Pakistan in the Beginning of the Pandemic: A Socio-Economic Analysis.
This study contributes to the existing literature on happiness studies by analyzing the effects of COVID-19 pandemic on subjective well-being (SWB) in a developing country, focusing specifically on satisfaction with socio-economic status. Drawing on survey data for urban Pakistan from before and after the outbreak of COVID-19, we find that during the early days of the pandemic and the related social distancing and potential lockdowns, SWB declined, particularly among unemployed, married couples, males and older people. Unexpectedly, we also observed that households having a higher income suffered more from the pandemic in terms of satisfaction with their socio-economic status compared to their poorer counterparts. We explain this finding by increased fear for falling into poverty due to lockdowns and inflation.


Sonographic findings in a case of acute cholecystitis with gallbladder perforation. A case report.
A case of acute cholecystitis with recent perforation is presented where the diagnosis was made preoperatively by ultrasound. The necessity to correlate the localization of the pain and the ultrasound findings is emphasized.


Direct determination of creatinine based on poly(ethyleneimine)/phosphotungstic acid multilayer modified electrode.
In this work, the direct determination of creatinine was achieved using a poly(ethyleneimine)/phosphotungstic acid multilayer modified electrode with the assistance of Copper(II) ions by cyclic voltammetry. The quantity of creatinine were determined by measuring the redox peak current of Cu(II)-creatinine complex/Cu(I)-creatinine complex. Factors affecting the response current of creatinine at the modified electrode were optimized. A linear relationship between the response current and the concentration of creatinine ranging from 0.125 to 62.5μM was obtained with a detection limit of 0.06μM. The proposed method was applied to determine creatinine in human urine, and satisfied results were gotten which was validated in accordance with high performance liquid chromatography. The proposed electrode provided a promising alternative in routine sensing for creatinine without enzymatic assistance.


[Therapeutic affect of zinc sulfate on central scotoma due to optic neuropathy of alcohol and tobacco abuse].
The oral administration of 400 mg/day for one month of zinc sulphate, when compared with placebo, provoked normal plasma zinc level in treated patients, principally men in their forties, with bilateral, retrobulbar, axial, optic neuropathy related to excessive smoking and consumption of ordinary wine and/or beer and/or spirits. Treatment significantly improved visual capacity (measured with Friedmann analyser) of the central scotoma area (5 degrees).


Probiotic treatment vs empiric oral antibiotics for managing dysbiosis in short bowel syndrome: Impact on the mucosal and stool microbiota, short-chain fatty acids, and adaptation.
Infants and children with short bowel syndrome (SBS) are presumed to be at risk of gut microbial dysbiosis with potential sequelae of bacterial overgrowth that include sepsis, d-lactic acidosis, mucosal inflammation, and malabsorption. In neonatal piglets with SBS, we compared intestinal microbial composition, short-chain fatty acids (SCFAs), and adaptation given probiotic (PRO) treatment (Lactobacillus and Bifidobacterium spp) vs oral metronidazole (MET). Following 75% distal small intestinal resection, piglets were allocated to PRO (500 mg twice a day, n = 7), MET (15 mg/kg twice a day, n = 8), and placebo (PLA) (500 mg twice a day, n = 8). After 10 days of parenteral and enteral nutrition, 16S ribosomal RNA gene amplicon sequencing (colon tissue and stool) was undertaken and SCFA analysis (stool and colon effluent) was performed using gas chromatography. In colon, Shannon diversity was higher for PRO compared with MET and PLA (P = 0.002). PRO and PLA increased abundance of Bacteroidetes species (eg, Bacteroides fragilis) compared with MET (P < 0.001). PRO, compared with PLA, increased abundance of Firmicutes species (eg, Lactobacillus fermentum) (P < 0.001). MET increased abundance of Proteobacteria members, predominately Enterobacteriaceae, compared with PRO (P = 0.004). In stool, microbial findings were similar and SCFA (butyrate) concentrations were highest for PRO (P = 0.003) compared with MET. In pediatric SBS, the empiric use of oral antibiotics, such as MET, is common for presumed clinical consequences of microbial dysbiosis. In this study of SBS piglets, that approach was associated with decreased microbial diversity and increased abundance of potentially inflammatory Proteobacteria. In contrast, a PRO treatment using Lactobacillus and Bifidobacterium spp increased both diversity and SCFAs.


Laparoscopic transabdominal preperitoneal hernioplasty for reduction en masse of an incarcerated inguinal hernia: a case report.
Reduction en masse is a rare complication of an incarcerated inguinal hernia. Its occurrence should be suspected when intestinal obstruction persists despite a seemingly successful manual reduction or hernioplasty. We report our experience in the management of a reduction en masse of a direct inguinal hernia. The diagnosis was established by computed tomography of the abdomen. The reduction en masse, as well as an accompanying indirect hernia, was successfully managed with laparoscopic transabdominal preperitoneal hernioplasty. The safety, effectiveness, and minimal invasiveness conferred by the laparoscopic approach justified its application under such conditions.


In a type 2 diabetic nephropathy rat model, the improvement of obesity by a low calorie diet reduces oxidative/carbonyl stress and prevents diabetic nephropathy.
The present study has been undertaken to unravel the critical factors involved in the progression of diabetic nephropathy (DN). A unique type 2 diabetic rat model with a wide range of metabolic derangements and hypertension has been utilized, the spontaneously hypertensive/NIH-corpulent rat SHR/NDmcr-cp(cp/cp). It develops histologically evident glomerular injury and tubulointerstitial damage, including mesangial activation, podocyte injury, and inflammatory cell infiltration in the tubulointerstitium. A low calorie diet for 22 weeks significantly improves obesity, proteinuria and renal morphological alterations. The correction of renal injury is independent of blood pressure control. Obesity correction, although partial, normalizes the renal content of pentosidine taken as a marker of oxidative stress and advanced glycation end products (AGEs). This occurs despite the fact that, in this model, improvement of glucose control and hyperlipidaemia is limited. Proteinuria and body weight are highly correlated with renal pentosidine content, while proteinuria and body weight are also correlated with each other. Diabetic renal injury is thus inhibited by a low calorie diet with an attendant reduction of oxidative stress and AGE formation, despite sustained hypertension. The present findings suggest a direct role of obesity in the generation of a localized oxidative stress and AGE formation, directly responsible for DN.


High-resolution melting analysis for SNP genotyping and mapping in tetraploid alfalfa (Medicago sativa L.).
Single nucleotide polymorphisms (SNPs) represent the most abundant type of genetic polymorphism in plant genomes. SNP markers are valuable tools for genetic analysis of complex traits of agronomic importance, linkage and association mapping, genome-wide selection, map-based cloning, and marker-assisted selection. Current challenges for SNP genotyping in polyploid outcrossing species include multiple alleles per loci and lack of high-throughput methods suitable for variant detection. In this study, we report on a high-resolution melting (HRM) analysis system for SNP genotyping and mapping in outcrossing tetraploid genotypes. The sensitivity and utility of this technology is demonstrated by identification of the parental genotypes and segregating progeny in six alfalfa populations based on unique melting curve profiles due to differences in allelic composition at one or multiple loci. HRM using a 384-well format is a fast, consistent, and efficient approach for SNP discovery and genotyping, useful in polyploid species with uncharacterized genomes. Possible applications of this method include variation discovery, analysis of candidate genes, genotyping for comparative and association mapping, and integration of genome-wide selection in breeding programs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11032-011-9566-x) contains supplementary material, which is available to authorized users.


Aqua mediated synthesis of substituted 2-amino-4H-chromenes and in vitro study as antibacterial agents.
A simple, clean, environmentally benign route to the synthesis of 2-amino-chromenes is described using K2CO3 as a green catalyst in water under microwave irradiation. This implies a convenient route avoiding the usage of hazardous organic solvents and organic bases. This technique requires only water in both the reaction step and workup, thus rendering the whole procedure into a truly ecofriendly green protocol. All the synthesized compounds were shown to possess antibacterial activity as tested in vitro against standard strains of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus.


Endocrine causes of hypertension in pregnancy--when to start looking for zebras.
Hypertension is a common medical disorder in pregnancy that may predate or first appear in pregnancy. Endocrine disorders rarely are the cause of the elevated blood pressure. However, it is essential to have a high index of suspicion because they carry much higher fetal and maternal morbidity and mortality risks. Endocrine disorders presenting as hypertension are primarily the result of autonomous production of renin, aldosterone, cortisol, or catecholamines. This report discusses the physiological changes in pregnancy, presentation, investigation, and management of these disorders.


Describing strong correlation with fractional-spin correction in density functional theory.
An effective fractional-spin correction is developed to describe static/strong correlation in density functional theory. Combined with the fractional-charge correction from recently developed localized orbital scaling correction (LOSC), a functional, the fractional-spin LOSC (FSLOSC), is proposed. FSLOSC, a correction to commonly used functional approximations, introduces the explicit derivative discontinuity and largely restores the flat-plane behavior of electronic energy at fractional charges and fractional spins. In addition to improving results from conventional functionals for the prediction of ionization potentials, electron affinities, quasiparticle spectra, and reaction barrier heights, FSLOSC properly describes the dissociation of ionic species, single bonds, and multiple bonds without breaking space or spin symmetry and corrects the spurious fractional-charge dissociation of heteroatom molecules of conventional functionals. Thus, FSLOSC demonstrates success in reducing delocalization error and including strong correlation, within low-cost density functional approximation.


Diffusion tensor imaging predicting neurological repair of spinal cord injury with transplanting collagen/chitosan scaffold binding bFGF.
Prognosis and treatment evaluation of spinal cord injury (SCI) are still in the long-term research stage. Prognostic factors for SCI treatment need effective biomarker to assess therapeutic effect. Quantitative diffusion tensor imaging (DTI) may become a potential indicators for assessing SCI repair. However, its correlation with the results of locomotor function recovery and tissue repair has not been carefully studied. The aim of this study was to use quantitative DTI to predict neurological repair of SCI with transplanting collagen/chitosan scaffold binding basic fibroblast growth factor (bFGF). To achieve our research goals, T10 complete transection SCI model was established. Then collagen/chitosan mixture adsorbed with bFGF (CCS/bFGF) were implanted into rats with SCI. At 8 weeks after modeling, implanting CCS/bFGF demonstrated more significant improvements in locomotor function according to Basso-Beattie-Bresnahan (BBB) score, inclined-grid climbing test, and electrophysiological examinations. DTI was carried out to evaluate the repair of axons by diffusion tensor tractgraphy (DTT), fractional anisotropy (FA) and apparent diffusion coefficient (ADC), a numerical measure of relative white matter from the rostral to the caudal. Parallel to locomotor function recovery, the CCS/bFGF group could significantly promote the regeneration of nerve fibers tracts according to DTT, magnetic resonance imaging (MRI), Bielschowsky's silver staining and immunofluorescence staining. Positive correlations between imaging and locomotor function or histology were found at all locations from the rostral to the caudal (P < 0.0001). These results demonstrated that DTI might be used as an effective predictor for evaluating neurological repair after SCI in experimental trails and clinical cases.


Endocrine and Neuroendocrine Tumors Special Issue-Checkpoint Inhibitors for Adrenocortical Carcinoma and Metastatic Pheochromocytoma and Paraganglioma: Do They Work?
Adrenocortical cancers and metastatic pheochromocytomas are the most common malignancies originating in the adrenal glands. Metastatic paragangliomas are extra-adrenal tumors that share similar genetic and molecular profiles with metastatic pheochromocytomas and, subsequently, these tumors are studied together. Adrenocortical cancers and metastatic pheochromocytomas and paragangliomas are orphan diseases with limited therapeutic options worldwide. As in any other cancers, adrenocortical cancers and metastatic pheochromocytomas and paragangliomas avoid the immune system. Hypoxia-pseudohypoxia, activation of the PD-1/PD-L1 pathway, and/or microsatellite instability suggest that immunotherapy with checkpoint inhibitors could be a therapeutic option for patients with these tumors. The results of clinical trials with checkpoint inhibitors for adrenocortical carcinoma or metastatic pheochromocytoma or paraganglioma demonstrate limited benefits; nevertheless, these results also suggest interesting mechanisms that might enhance clinical responses to checkpoint inhibitors. These mechanisms include the normalization of tumor vasculature, modification of the hormonal environment, and vaccination with specific tumor antigens. Combinations of checkpoint inhibitors with classical therapies, such as chemotherapy, tyrosine kinase inhibitors, radiopharmaceuticals, and/or novel therapies, such as vaccines, should be evaluated in clinical trials.


Innershell absorption spectroscopy of amino acids at all relevant absorption edges.
The C, N, and O K-edge near-edge X-ray absorption fine structure spectra of the 22 most common proteinogenic alpha-amino acids in the zwitterionic form collected from solvent-free polycrystalline powder films in the partial electron yield mode are reported. Spectral features common to all amino acids, as well as distinctive fingerprints of specific subgroups of these compounds, are presented and discussed.


Performance changes in the anammox process under the stress of rare-earth element Ce(III) and the evolution of microbial community and functional genes.
The high Ce(III) content in ionic rare-earth tailings wastewater has hindered the application of anammox process in this field. Here, the effect of Ce(III) on the performance of anammox processes was investigated, and the evolution of microbial communities and functional genes was explored using metagenomic sequencing. The results showed that the reactor nitrogen removal rate decreased when the Ce(III) concentration reached 25 mg/L, although ammonia nitrogen removal (92.31%) and nitrogen removal efficiency (81.33%) remained at a high level; however, both showed a significant decreasing trend. The relative abundance of anammox bacteria increased continuously from P1-P5, reaching 48.81%, whereas the relative abundance of Candidatus jettenia reached 33.71% at P5, which surpassed that of Candidatus brocadia as the most abundant anammox bacteria, and further analysis of functional genes and metabolic pathways revealed that Candidatus brocadia was richer in biochemical metabolic genes, whereas Candidatus jettenia had richer efflux genes.


Grey matter correlates of affective and somatic symptoms of premenstrual dysphoric disorder.
Ovarian hormones fluctuations across the menstrual cycle are experienced by about 58% of women in their fertile age. Maladaptive brain sensitivity to these changes likely leads to the severe psychological, cognitive, and physical symptoms repeatedly experienced by women with Premenstrual Dysphoric Disorder (PMDD) during the late luteal phase of the menstrual cycle. However, the neuroanatomical correlates of these symptoms are unknown. The relationship between grey matter structure and PMDD symptom severity was delineated using structural magnetic resonance imaging during the late luteal phase of fifty-one women diagnosed with PMDD, combined with Voxel- and Surface-Based Morphometry, as well as subcortical volumetric analyses. A negative correlation was found between depression-related symptoms and grey matter volume of the bilateral amygdala. Moreover, the severity of affective and somatic PMDD symptoms correlated with cortical thickness, gyrification, sulcal depth, and complexity metrics, particularly in the prefrontal, cingulate, and parahippocampal gyri. The present findings provide the first evidence of grey matter morphological characteristics associated with PMDD symptomatology in brain regions expressing ovarian hormone receptors and of relevance to cognitive-affective functions, thus potentially having important implications for understanding how structural brain characteristics relate to PMDD symptomatology.


ENA Actimineral Resource A restores bone loss and bone quality in ovariectomized rats.
The purpose of this study was to examine the effects of ENA Actimineral Resource A (ENA-A), seaweed origin alkaline water, on postmenopausal osteoporosis in ovariectomized (OVX) rats. The 12-week old Wistar rats were divided randomly into 4 groups: ovariectomized (OVX), OVX plus 0.5% ENA-A, OVX plus 5% ENA-A and OVX plus 10% ENA-A. A histopathological analysis indicated that ENA-A could prevent OVX-induced bone loss by increasing femur trabecular bone area in a dose-dependent manner. ENA-A significantly (p<0.05) increased serum estradiol levels, decreased serum osteocalcin activity and suppressed serum pyridinoline (PYD) levels. The in vitro effects of ENA-A were also studied using MC3T3-E1 cells. ENA-A significantly stimulated cell proliferation and increased both ALP activity and calcium deposition in a dose-dependent manner. These results suggest that the treatment of ovariectomized rats with ENA-A not only prevents bone resorption but also appears to maintain the cancellous bone structure of postmenopausal osteoporosis.


The ovine conceptus utilizes extracellular serine, glucose, and fructose to generate formate via the one carbon metabolism pathway.
Highly proliferative cells rely on one carbon (1C) metabolism for production of formate required for synthesis of purines and thymidine for nucleic acid synthesis. This study was to determine if extracellular serine and/or glucose and fructose contribute the production of formate in ovine conceptuses. Suffolk ewes (n = 8) were synchronized to estrus, bred to fertile rams, and conceptuses were collected on Day 17 of gestation. Conceptuses were either snap frozen in liquid nitrogen (n = 3) or placed in culture in medium (n = 5) containing either: 1) 4 mM D-glucose + 2 mM [U-13C]serine; 2) 6 mM glycine + 4 mM D-glucose + 2 mM [U-13C]serine; 3) 4 mM D-fructose + 2 mM [U-13C]serine; 4) 6 mM glycine + 4 mM D-fructose + 2 mM [U-13C]serine; 5) 4 mM D-glucose + 4 mM D-fructose + 2 mM [U-13C]serine; or 6) 6 mM glycine + 4 mM D-glucose + 4 mM D-fructose + 2 mM [U-13C]serine. After 2 h incubation, conceptuses in their respective culture medium were homogenized and the supernatant analyzed for 12C- and 13C-formate by gas chromatography and amino acids by high performance liquid chromatography. Ovine conceptuses produced both 13C- and 12C-formate, indicating that the [U-13C]serine, glucose, and fructose were utilized to generate formate, respectively. Greater amounts of 12C-formate than 13C-formate were produced, indicating that the ovine conceptus utilized more glucose and fructose than serine to produce formate. This study is the first to demonstrate that both 1C metabolism and serinogenesis are active metabolic pathways in ovine conceptuses during the peri-implantation period of pregnancy, and that hexose sugars are the preferred substrate for generating formate required for nucleotide synthesis for proliferating trophectoderm cells.


A novel route for waste water treatment: photo-assisted Fenton degradation of dye pollutants accumulated in natural polyelectrolyte microshells.
The efficient accumulation of dyes in constructed natural polyelectrolyte microshells under moderate conditions, combined with the photo-assisted Fenton reagent, opens a new route for the effective elimination of dye pollutants from waste water.


Identification of the novel HLA-A*23:01:01:27 allele in an acute myeloid patient and related donor.
A single nucleotide polymorphism in intron 1 determines the new HLA-A*23:01:01:27 allele.


Is rectal washout effective for preventing localized recurrence after anterior resection for rectal cancer?
The present study evaluated the effect of rectal washout in reducing local recurrence after resection for rectal cancer. A literature search was performed on studies published since 1989 that compared rectal washout to no washout for rectal cancer resection. Primary end point was local cancer recurrence. Random-effect meta-analysis was used and subgroup analysis was performed. Five studies matched the selection criteria, and reported on 176 patients who underwent rectal washout and 256 who did not undergo washout. Different washout solutions were used in every study, and total mesorectal excision was not universally applied. Overall local recurrence rate was 8 percent (33/432). Local recurrence rate for rectal washout patients was 4.8 percent compared with 10.2 percent for patients who did not undergo rectal washout, a difference that was not statistically significant (odds ratio = 0.64; 95 percent confidence interval = 0.2-2.04). When only studies using total mesorectal excision were considered, there was no significant difference between the two groups (odds ratio = 1.21; 95 percent confidence interval = 0.37-3.92). Although no definitive conclusions may be drawn because of the nonrandomized nature of the included studies, rectal washout is relatively risk-free and adds little to the operative time. This may be performed until a randomized, controlled trial is undertaken to resolve this contentious issue.


Childhood vulval lichen sclerosus in a patient with ectodermal dysplasia and uncombable hair.
An 11-year-old girl was diagnosed as having ectodermal dysplasia, with abnormalities of her nails, teeth, and hair noted from infancy. Her hair was unruly and investigations confirmed the changes seen in "cheveux incoiffables"-the uncombable hair syndrome. She also had atopy requiring treatment to control asthma and eczema. When she was 8 years old she developed vulval lichen sclerosus, which was controlled easily with early treatment. The coexistence of these conditions is discussed.


Proximity labeling identifies LOTUS domain proteins that promote the formation of perinuclear germ granules in C. elegans.
The germ line produces gametes that transmit genetic and epigenetic information to the next generation. Maintenance of germ cells and development of gametes require germ granules-well-conserved membraneless and RNA-rich organelles. The composition of germ granules is elusive owing to their dynamic nature and their exclusive expression in the germ line. Using Caenorhabditis elegans germ granule, called P granule, as a model system, we employed a proximity-based labeling method in combination with mass spectrometry to comprehensively define its protein components. This set of experiments identified over 200 proteins, many of which contain intrinsically disordered regions (IDRs). An RNA interference-based screen identified factors that are essential for P granule assembly, notably EGGD-1 and EGGD-2, two putative LOTUS-domain proteins. Loss of eggd-1 and eggd-2 results in separation of P granules from the nuclear envelope, germline atrophy, and reduced fertility. We show that IDRs of EGGD-1 are required to anchor EGGD-1 to the nuclear periphery while its LOTUS domains are required to promote the perinuclear localization of P granules. Taken together, our work expands the repertoire of P granule constituents and provides new insights into the role of LOTUS-domain proteins in germ granule organization.


Rapid progression of cervical squamous cell carcinoma with delayed treatment in pregnancy.
•Delayed treatment of cervical cancer in pregnancy can result in progression.•Surveillance of cervical cancer in pregnancy with pelvic MRIs every 6 weeks.•Comprehensive multidisciplinary care is essential in setting of treatment delays.


Integrins in mammary gland development and differentiation of mammary epithelium.
Integrins are major extracellular matrix (ECM) receptors that can also serve for some cell-cell interactions. They have been identified as important regulators of mammary epithelial cell growth and differentiation. Their ability to promote cell anchorage, proliferation, survival, migration, and the induction of active ECM-degrading enzymes suggests that they play an essential role in normal mammary morphogenesis, but, on the other hand, reveals their potential to promote tumor progression.


Orthogonal protein purification--expanding the repertoire of GST fusion systems.
We have previously developed a labeling scheme that can be used to site-specifically link human glutathione transferases (hGSTs) from the alpha class to chemical entities such as fluorophores and aldehydes. The reagents are in-house synthesized derivatives of glutathione (GS-derivatives). We have focused on a lysine mutant of hGST A1:A216K. In this study, we wanted to utilize these findings and improve on protein purification schemes that are using GSTs as fusion partners. We have used random mutagenesis to scramble the hydrophobic binding site of A216K through mutations at position M208 and isolated a library of 11 A216K/M208X mutants. All mutants were easily expressed and purified and retained all or parts of the catalytic properties of the parent GST. The mutants were stable over several days at room temperature. The A216K/M208X mutants could be site-specifically labeled using our designed fluorescent reagents. Furthermore, reaction with an aldehyde-containing reagent termed GS-Al results in site-specific introduction of an orthogonal handle for subsequent conjugation with aldehyde-reactive probes. Labeling with coumarin results in a fluorescent protein-conjugate that can bind glutathione (GSH) derivatives for subsequent affinity purification. The K(d) for S-hexyl-GSH of coumarin-labeled A216K was measured to be 2.5 microM. The candidate proteins A216K and A216K/M208F could be purified in high yield in a one-step procedure through affinity chromatography (Glutathione Sepharose 4B). The proteins can readily be perceived as improved GST fusion partners.