Uterine and placenta characteristics during early vascular development in the pig from day 22 to 42 of gestation.
Insufficient placenta development is one of the primary causes of fetal death and reduced fetal growth after 35 days of gestation. Between day 22 and 42 the placenta consists of a central highly vascular placenta (HVP), adjacent to the fetus, a less vascular placenta (LVP), on either side of the fetus, and necrotic tips (NT). The objective of this study was to comprehensively evaluate uterine-placenta characteristics during early gestation in the gilt and determine time points and physiological changes. Gilts (n=25) were artificially inseminated at first detection of estrus (day 0) and 24h later, and harvested at 22, 27, 32, 37 or 42 days of gestation. Litter size, 12.1±3.4, was similar for all days of gestation. Fetal and placenta weight increased with day of gestation. The greatest increase in placenta weight occurred between 37 and 42 days of gestation. The LVP zones had no measurable fold formation until day 27. Necrotic tips became apparent after 27 days of gestation. Unoccupied areas of the uterus developed folds with changes in endometrial cell size and morphology from day 32 to 42 of gestation. Limited changes occurred in either fetal growth or placenta weight from day 27 through 32 of gestation; however, significant morphological changes occur at the maternal-fetal interface, demonstrating the dynamic architecture of the developing porcine placenta during early gestation. This work establishes fundamental time points in placenta development corresponding to fetal growth and microfold formation that may influence fetal growth and impact fetal survival.


Detection of the granulocyte-specific antigen 3-fucosyl-N-acetyl-lactosamine on leukemic cells after neuraminidase treatment.
The carbohydrate antigen 3-fucosyl-N-acetyl-lactosamine (FAL) is expressed on human granulocytes and is detected by a monoclonal antibody B4.3. After neuraminidase treatment, this structure can also be detected on monocytes and on the cells of nearly all acute myeloid leukemia patients (38/39). It is then also present on the cells of a number of CALLA-positive lymphatic leukemias (8/18), but not on T-ALL and B-ALL cells. On cells of patients with AUL, the antigen is then detected in many TdT+ cases, but not in TdT- cases.


Complete Genome Sequence of Caulobacter crescentus Siphophage Sansa.
Caulobacter crescentus is a Gram-negative dimorphic model organism used to study cell differentiation. Siphophage Sansa is a newly isolated siphophage with an icosahedral capsid that infects C. crescentus. Sansa shares no sequence similarity to other phages deposited in GenBank. Here, we describe its genome sequence and general features.


Effective high-density lipoprotein cholesterol is associated with carotid intima-media thickness and vascular events after acute ischemic stroke.
Effective high density lipoprotein cholesterol (HDL-C) is a measure of HDL functionality. We evaluated if HDL-C is associated with carotid intima-media thickness (cIMT) and incident major adverse cardiovascular events (MACE) in patients with acute ischemic stroke from two prospective cohort studies. In the MARK-STROKE cohort, 299 patients with acute ischemic stroke or TIA were included. Outcome was available in 219 patients during a median follow-up of 294 days. In CIRCULAS, 382 acute ischemic stroke patients were included and a 90-day follow-up was available in 213 patients. HDL-C was calculated based on symmetric dimethylarginine (SDMA) and HDL cholesterol concentrations. Main outcome was incident MACE (death, stroke, and myocardial infarction) and the main measure was cIMT. In both studies, HDL-C was inversely associated with cIMT in linear regression analysis adjusted for age, sex and creatinine. In MARK-STROKE, the adjusted hazard for MACE was significantly reduced for patients with one unit increase (mg/dL) of HDL-C (hazard ratio 0.95 [95% confidence interval (CI): 0.92, 0.99]). In the CIRCULAS cohort, stroke patients with higher HDL-C had less incident MACE during 90 days of follow-up (odds ratio: 0.97 [95% CI: 0.94, 0.99] for one unit increase). Neither SDMA nor HDL cholesterol predicted outcome. Our findings imply a protective role of biologically effective HDL after acute cerebral ischemia for secondary events and emphasize the relevance of lipoprotein functionality in these patients.


Uptake and axonal transport of horseradish peroxidase isoenzymes by different neuronal types.
The uptake and transport of basic and acidic horseradish peroxidase isoenzymes was compared in the neuromuscular, visual and olfactory systems of Xenopus larvae and postmetamorphic frogs. The concentration (w/v) of the two preparations was corrected to compensate for their difference in enzymatic activity (unit/w), which was seven-fold higher in basic horseradish peroxidase. Uptake and transport of horseradish peroxidase isoenzymes could be demonstrated with 7% basic horseradish peroxidase, but not with equal amounts of 49% acidic horseradish peroxidase in all systems investigated: retrograde transport from terminals of retinal ganglion cells, isthmotectal neurons and spinal motoneurons, as well as anterograde transganglionic transport along olfactory neurons. A very weak labelling of the same neuronal pathways by acidic horseradish peroxidase was obtained only after increasing the amount injected by approximately two-fold. Basic horseradish peroxidase isoenzymes were also preferentially taken up and transported retrogradely by broken axons of the optic nerve. When tested, similar results were obtained in both larvae and frogs suggesting that preferential uptake and transport of basic horseradish peroxidase is a general feature of all neurons and of all developmental stages. Electron microscopical analysis of the outer layers of the optic tectum revealed that, in the same experimental conditions producing no retrotrade labelling of optic axons, acidic horseradish peroxidase was rarely found to enter nerve terminals. It appears that interactions between horseradish peroxidase and neuronal membranes occur during uptake and transport and that molecular charge plays an important role, beyond non-specific fluid-phase endocytosis. We suggest that differences between horseradish peroxidase isoenzymes as neuronal tracers reflect a process of adsorptive endocytosis related to general characteristics of neuronal membranes (regardless of age) and not to specific receptor-mediated interactions characteristic of neuronal specificity.


Simultaneous ingestion of dietary proteins reduces the bioavailability of galloylated catechins from green tea in humans.
To investigate the influence of dietary proteins (casein, soy protein) and skimmed milk on the plasma kinetics of green tea (GT) catechins. In a randomized cross-over design with one-week intervals, 24 healthy normal-weight women consumed a test drink containing 1.75 g GT extract with or without the addition of different proteins. Treatments were GT (control), GT with skimmed milk (GT + M), GT with caseinate (GT + CS), or GT with soy protein (GT + S). Venous blood samples were taken before and several times during a period of 4.5 h after consumption of the test drink. Plasma concentrations of catechins were analyzed by HPLC with electrochemical detection. Compared to control, consumption of GT with milk, caseinate, or soy protein significantly reduced the bioavailability (mean area under the plasma concentration-time curve) of total catechins (means ± SEM; GT + M, 87 ± 5%; GT + CS, 79 ± 5%; GT + S, 88 ± 4%), epigallocatechin gallate (GT + M, 68 ± 4%; GT + CS, 63 ± 5%; GT + S, 76 ± 5%), and epicatechin gallate (GT + M, 68 ± 5%; GT + CS, 66 ± 6%; GT + S, 77 ± 6%), while the bioavailability of non-galloylated catechins such as epigallocatechin (GT + M, 134 ± 9%; GT + CS, 118 ± 9 %; GT + S, 123 ± 8%) and epicatechin (GT + M, 125 ± 10%; GT + CS, 114 ± 11%; GT + S, 110 ± 8%) significantly increased. No significant differences in bioavailability of GT catechins were observed between the treatments GT + M, GT + CS, or GT + S. Simultaneous ingestion of dietary proteins reduces the bioavailability of galloylated catechins from GT in humans.


Galectin-3 suppresses mucosal inflammation and reduces disease severity in experimental colitis.
Galectin-3, a member of the β-galactoside-binding lectin family, expresses in many different immune cells and modulates broad biological functions including cell adhesion, cell activation, cell growth, apoptosis, and inflammation. However, the role of galectin-3 in mucosal immunity or inflammatory bowel diseases is still not clear. We demonstrate here that galectin-3 knockout mice have more severe disease activity in the dextran sulfate sodium (DSS)-induced colitis model, indicating that galectin-3 may protect from inflammation in DSS-induced colitis. Furthermore, treating with galectin-3 reduced body weight loss, shortened colonic length, and ameliorated mucosal inflammation in mice having DSS-induced colitis. However, the protective effects of galectin-3 were eliminated by the administration of anti-CD25 mAb. In addition, primary T cells treated with galectin-3 ex vivo induced the expression of FOXP3, ICOS, and PD-1 with a Treg cell phenotype having a suppression function. Moreover, adoptive transfer of galectin-3-treated T cells reduced bowel inflammation and colitis in the T cell transfer colitis model. In conclusion, our results indicate that galectin-3 inhibited colonic mucosa inflammation and reduced disease severity by inducing regulatory T cells, suggesting that it is a potential therapeutic approach in inflammatory bowel disease. Galectin-3 offers protection from inflammation in experimental colitis. Galectin-3 knockout mice have more severe disease activity in DSS-induced colitis. Adoptive transfer of galectin-3-treated T cells reduced bowel inflammation. Galectin-3 inhibited colonic mucosa inflammation by inducing regulatory T cells. Galectin-3 is a potential therapeutic approach in inflammatory bowel disease.


Design of MEMS devices with optical apertures for the detection of transparent biological cells.
This paper provides a novel technique to detect transparent biological living cells trapped in a microfluidic MEMS device by optical diffraction. The device essentially consists of an optical aperture or an aperture array patterned in metal layer and a microfluidic chamber positioned above the center of the aperture. When the cells in the chamber are illuminated through the aperture, the far-field diffraction pattern can be recorded by a CCD camera or a photodetector array. This diffraction pattern uniquely corresponds to the sizes, positions, and intrinsic optical properties of the aperture, cells, and the microfluidic chamber materials, so any unknown but relevant parameter is able to be extrapolated when all other parameters are fixed or identified. This paper describes in detail the designs of various microfluidic chambers and apertures for this application, and the development of a complete set of software for the analysis of the cells' optical properties. Compared with other currently available methods for the detection of transparent living cells, this method has the advantages of simple device structure, easy to manipulate, able to simultaneously detect several cells of different species, as well as providing accurate and sensitive results. Besides the detection of living cells, this technique can also be used to detect or characterize other transparent or low optical absorption particles, such as polymer spheres or insoluble droplets, inside an aqueous solution.


Immunotherapy in Acute Leukemias: Past Success Paves the Way for Future Progress.
Immunotherapy as a cancer treatment modality has undergone recent widespread proliferation across all cancer types, especially amongst patients with solid tumors. However, the longest tenured immunotherapy approach to cancer is allogeneic stem cell transplantation (allo-SCT) for two hematologic malignancies: acute myeloid and acute lymphoid leukemia (AML and ALL, respectively). While allo-SCT remains a standard of care for eligible patients, recent advances/applications of monoclonal antibodies, immune checkpoint inhibitors, bispecific T-cell engagers (BiTEs), and CAR T-cell therapy are changing the treatment landscape for these acute leukemias by either direct to tumor immune targeting or through decreased toxicities that expand patient eligibility. Pre-clinical data and clinical trials have shown promising results for novel immunotherapies in acute leukemia, and multiple ongoing trials are investigating these novel approaches. While there have been promising results with these approaches, particularly in the relapsed/refractory setting, there remain challenges in optimizing the use of these therapies, such as managing cytokine release syndrome and other immune-related toxicities. Immunotherapy is a rapidly evolving field in the treatment of acute leukemia and has the potential to significantly impact the management of both AML and ALL. This review highlights the history of immunotherapy in the treatment of acute leukemias, the evolution of immunotherapy into more targeted approaches, the potential benefits and limitations of different immune targeting approaches, and ongoing research and development in the field.


Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes.
Six simple methods for extraction of ribosomal and mitochondrial DNA from Toxocara canis, Toxocara cati and Toxascaris leonina were compared by evaluating the presence, appearance and intensity of PCR products visualized on agarose gels and amplified from DNA extracted by each of the methods. For each species, two isolates were obtained from the intestines of their respective hosts: T. canis and T. leonina from dogs, and T. cati from cats. For all isolates, total DNA was extracted using six different methods, including grinding, boiling, crushing, beating, freeze-thawing and the use of a commercial kit. To evaluate the efficacy of each method, the internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit 1 (cox1) gene were chosen as representative markers for ribosomal and mitochondrial DNA, respectively. Among the six DNA extraction methods, the beating method was the most cost effective for all three species, followed by the commercial kit. Both methods produced high intensity bands on agarose gels and were characterized by no or minimal smear formation, depending on gene target; however, beating was less expensive. We therefore recommend the beating method for studies where costs need to be kept at low levels.


A tungsten-in-glass iontophoresis assembly for studying input-output relationships in central neurons.
A method is described for the production of an electrode capable of monitoring and modulating the input-output relationship of thalamic neurons. Tungsten-in-glass electrodes were manufactured with the ability to simultaneously record lateral geniculate nucleus action potentials and associated, retinally evoked S-potentials. The recording electrodes were mounted onto multibarreled micropipettes with iontophoretic capability. The completed electrode assembly permitted micropharmacological modulation of the fraction of lateral geniculate nucleus output spikes to retinal input spikes (the transfer ratio). Iontophoretically applied gamma-aminobutyric acid (GABA) decreased the transfer ratio, an effect countered by the GABAA antagonist, bicuculline. Elevated transfer ratios produced by stimulation of an afferent pathway originating in the parabrachial region of the brainstem were decreased by concurrently applied GABA. The fabrication of this electrode assembly employs simple modifications of existing techniques and separate construction of recording and iontophoretic elements to provide high-quality single-unit recordings coupled with micropharmacological function.


In vitro coordination of Fe-protoheme with amyloid β is non-specific and exhibits multiple equilibria.
In addition to copper and zinc, heme is thought to play a role in Alzheimer's disease and its metabolism is strongly affected during the course of this disease. Amyloid β, the peptide associated with Alzheimer's disease, was shown to bind heme in vitro with potential catalytic activity linked to oxidative stress. To date, there is no direct determination of the structure of this complex. In this work, we studied the binding mode of heme to amyloid β in different conditions of pH and redox state by using isotopically labelled peptide in combination with advanced magnetic and vibrational spectroscopic methods. Our results show that the interaction between heme and amyloid β leads to a variety of species in equilibrium. The formation of these species seems to depend on many factors suggesting that the binding site is neither very strong nor highly specific. In addition, our data do not support the currently accepted model where a water molecule is bound to the ferric heme as sixth ligand. They also exclude structural models mimicking a peroxidatic site in the amyloid β-Fe-protoheme complexes.


The expanding spectrum of HCV-related cryoglobulinemic vasculitis: a narrative review.
Cryoglobulinemic vasculitis (CV) is a small-to-medium-vessel vasculitis that appears in 10-15 % of patients chronically infected with hepatitis C virus (HCV). The classic symptom triad of CV, purpura/asthenia/arthralgia, is accompanied by clinical features that include glomerulonephritis, neuropathy, interstitial pneumonitis, and cardiomyopathy, ranging in their severity from mild to life threatening. The risk of developing non-Hodgkin lymphoma is also higher. The cumulative 10-year survival rate of CV patients is significantly lower than in the age- and sex-matched general population, with death typically caused by nephropathy, malignancies, liver involvement, and severe infections. Unfailing serological stigmata include both a cryoglobulin IgM fraction with rheumatoid factor activity and decreased complement C4 levels. On peripheral B cells, the expression of the CD81 B cell receptor is reduced while that of the CD19 receptor is increased. A monoclonal B cell lymphocytosis develops in almost one-third of patients. HCV-related proteins (but not HCV-RNA genomic sequences) can be detected on biopsy samples by immunofluorescence and immunohistochemistry and involve the vessel lumen, vessel walls, and the perivascular spaces of the skin, kidney, and peripheral nerves, supporting the pathogenetic role of HCV in the onset of a widespread microvasculitis. Based on the demonstration of HCV infection in the large majority of CV patients, a therapeutic regimen consisting of once-weekly pegylated interferon-α and the daily administration of ribavirin results in a sustained virologic response in ~50 % of patients. In those with refractory and relapsing disease, addition of the anti-CD20 monoclonal antibody rituximab has significantly increased the overall response rates. The extension to CV of latest-generation direct-acting antivirals, strikingly successful in non-CV HCV-positive patients, has yielded high complete response rates according to the few studies published thus far.


Expression of post-transcriptional regulatory gene of HTLV-I, rex, in Escherichia coli.
Human T-cell leukemia virus type-I (HTLV-I) has a post-transcriptional regulatory gene termed rex. We have designed the rex gene to express in E. coli. Synthesis of rex protein, p27rex, was examined by immunoblot analysis using anti-p27rex antibody. No difference in electrophoretic mobility in NaDadSO4-PAGE was observed between p27rex expressed in E. coli and in an HTLV-I-infected cell line, MT-2. Slower migration of p27rex, corresponding to a 27-kD protein, in NaDodSO4-PAGE when compared with the calculated molecular weight from the amino acid sequence (Mr = 20,367) is suggested to be caused not by post-translational modification, but by the intrinsic nature of the protein, which is rich in proline and arginine.


Design, synthesis of 8-alkoxy-5,6-dihydro-[1,2,4]triazino[4,3-a]quinolin-1-ones with anticonvulsant activity.
A new series of 8-alkoxy-5,6-dihydro-[1,2,4]triazino[4,3-a]quinolin-1-one derivatives were synthesized. Their anticonvulsant activities were evaluated by the maximal electroshock (MES) test, and their neurotoxicities were evaluated by the rotarod neurotoxicity test. The results showed that 8-heptyloxy-5,6-dihydro-[1,2,4]triazino[4,3-a]quinolin-1-one 5t was the most potent with median effective dose (ED(50)) value of 11.4 mg/kg, median toxicity dose (TD(50)) of 114.1 mg/kg, providing a protective index (PI=TD(50)/ED(50)) value of 10.0, which is much greater than the PI of the prototype drug carbamazepine (PI=6.4). To explain the possible mechanism of anticonvulsant activity, the compound 5t was tested in chemically induced seizures.


A systematic review of preceptor's experience in supervising undergraduate nursing students: Lessons learned for mental health nursing.
Preceptorship in nursing has been a valued concept in nursing. Speciality area such as mental health nursing has a massive gap in research study. To develop sturdy mental health nursing workforce, it is necessary to conduct more studies. This literature review aims to explore preceptor's experience in precepting undergraduate nursing students in mental health. Systematic review of literature. The systematic review was conducted from January 2021 to August 2021. Population of the studies included Registered Nurses supervising nursing students in the clinical area. Only studies conducted in English were included. A systematic search using EBSCO Host databases, CINAHL Complete, MEDLINE APA Psycinfo & APA PsycArticles, has been used in this review paper. Papers were also selected from the citation reference of included papers. The new version of the PRISMA 2020 guidelines used to represent the process of selection of papers has been incorporated as part of this review. The final set of data included 14 original papers meeting the eligibility criteria which involved quantitative (n = 5), qualitative (n = 4) and mixed-method studies (n = 5). Results were presented under three major themes: time-consuming, lack of recognition and need extra support. Further research is required in the mental health clinical setting to effectively explore the impact of relationships between preceptors and the preceptees. Preceptors reported supervising students in the clinical area has many benefits. However, some challenges they raised were increase in workload, requiring some guidance and acknowledgement from the organization.


[Refractive predictability and stability of three-piece versus single-piece intraocular lenses in patients with high axial myopia].
Based on previous data on single-piece and three-piece intraocular lenses (IOLs) there is no evidence for significant differences in decentration, tilt and refractive shift. The purpose of the current study was to compare single-piece and three-piece IOLs in patients with high axial myopia. A total of 68 eyes of 50 patients with high axial myopia (axis length ≥ 28.00 mm) with and without cataract who underwent complication-free phacoemulsification and IOL implantation were retrospectively examined. To compare single-piece and three-piece IOLs, the patients were retrospectively grouped depending on IOL type: group 1 acrylic single-piece IOL (n = 37; ACR6D SE, Corneal, France) and group 2 acrylic three-piece IOL with fixed haptic frame (n = 31; AF-1 UY, Hoya, Japan). Patient files were analyzed regarding best spectacle-corrected visual acuity (BSCVA), refractive predictability and stability. In this study the mean BSCVA was determined as 0.22 ± 0.12 logMAR and 0.13 ± 0.11 logMAR 6 months postoperatively for the ACR6D SE group and the AF-1 UV group, respectively (p = 0.09). Refractive predictability was less accurate in the ACR6D SE (+ 1.75 ± 2.2 dpt) compared to the AF-1 UV (- 0.37 ± 1.1) treated eyes (p = 0.001). Refractive stability, defined as the difference in diopters between the first week and the sixth month after surgery, resulted in + 0.40 ± 1.7 dpt and -0.16 ± 1.2 dpt for the ACR6D SE and the AF-1 UV, respectively (p = 0.022). The three-piece AF-1 UV showed satisfactory refractive predictability and stability in patients with high axial myopia. The ACR6D SE has a high refractive unpredictability and should not be used in eyes with high axial myopia.


Potential role for adenosine in the pathogenesis of the vascular complications of hyperhomocysteinemia.
Hyperhomocysteinemia is an independent risk factor for cardiovascular disease. Most previous investigations focused on the role of homocysteine as direct pathogenetic factor for these adverse vascular events. However, the exact pathophysiological mechanism is still unknown. In this review we discuss the hypothesis that a decreased extracellular concentration of adenosine could contribute to the adverse cardiovascular effects of hyperhomocysteinemia. Fundamental to this hypothesis is that, in vivo, any increase in the plasma concentration of homocysteine reflects an increased intracellular homocysteine concentration, which inevitably will result in a decrease in the adenosine concentration. In this situation, the hydrolase reaction catalysed by S-adenosylhomocysteine hydrolase will reverse and S-adenosylhomocysteine will accumulate at the expense of adenosine. Stimulation of adenosine receptors by adenosine results in various cardio- and vasoprotective actions, like modulation of vascular resistance, presynaptic inhibition of norepinephrine release, ischaemic preconditioning, inhibition of platelet aggregation, modulation of inflammation and regulation of vascular cell proliferation and death. In this respect, a decrease in the adenosine concentration could contribute significantly to the cardiovascular effects of hyperhomocysteinemia.


Natural experiments for orthopaedic trauma research: An introduction.
Natural experiments are observational studies of medical treatments in which treatment allocation is determined by factors outside the control of the investigators, arguably resembling experimental randomisation. Natural experiments in the field of orthopaedic trauma research are scarce. However, they have great potential due to the process governing treatment allocation and the existence of opposing treatment strategies between hospitals or between regions as a result of local education, conviction, or cultural and socio-economic factors. Here, the possibilities and opportunities of natural experiments in the orthopaedic trauma field are discussed. Potential solutions are presented to improve the validity of natural experiments and how to assess the credibility of such studies. Above all, it is meant to spark a discussion about its role within the field of orthopaedic trauma research.


Pediatric Cardiology Condolence Letter Writing: Does a Fellowship Curriculum Impact Practice?
Condolence letter (CL) writing after the death of a child is an important opportunity for humanism. Pediatric cardiology fellowship training now recognizes the importance of palliative care, but rarely includes CL education, despite its fragile patient population. To address this professionalism gap, a formal CL writing curriculum was created and implemented in a pediatric cardiology fellowship. This study investigated the impact of the curriculum on pediatric cardiology CL writing, and broader CL practices and beliefs. Pediatric cardiology fellows at a high volume urban academic program from 2000 to 2022 were divided into two cohorts (exposure to CL curriculum [2014-2022] vs. no exposure [2000-2013]) and responded by anonymous electronic multiple choice and open ended survey to assess the CL curriculum and describe current CL practices and beliefs. Impact of curriculum elements was determined by ordinal ranking. A 5-point Likert scale was used to report physician behaviors. Chi-square tests of independence were utilized for group comparisons. The overall survey response rate was 59% (63/107). Cardiologists who participated in the curriculum (64%, 35/55) were more likely to report writing CLs (80% vs. 40%; P < 0.01). Impactful curriculum elements included the opportunity for all fellows to contribute to a CL (78%) and identifying a primary fellow to write the CL (66%). A majority (>75%) of curriculum participants agreed that formal teaching increased their frequency, ability, and comfort in writing CLs. Development of condolence expression educational programs in pediatric cardiology training should be expanded.


Mortality in older patients admitted to an ICU for COVID-19: A systematic review.
The objective was to conduct a systematic review of mortality and factors independently associated with mortality of older patients admitted to an intensive care unit (ICU) for COVID-19. Data sources were MEDLINE, EMBASE, the Cochrane Library, and references of included studies. Two reviewers independently selected studies evaluating mortality of older patients (≥ 70 years) admitted to an ICU for COVID-19. They extracted general characteristics, mortality rate, and factors independently associated with mortality. The methodological quality of each study was evaluated by using the Critical Appraisal Skills Programme checklist. We selected 36 studies (11,989 patients). Many of the studies were conducted in Europe (42%) and many were retrospective (61%) and multicenter (61%). ICU mortality ranged from 8% to 90%, 1-month mortality from 33% to 90% and 3-month mortality, reported in five studies, from 46% to 60%. Frailty, assessed by the Clinical Frailty Score (CFS), was significantly associated with 1-month and 3-month mortality respectively in two studies (hazard ratio [HR]: 3.2 [2.56-4.13] and HR: 2.83 [95% CI: 1.96-4.08]). In this systematic review of older patients admitted to an ICU with COVID-19, we documented high heterogeneity of mortality rates.


Bcr-Abl dependent post-transcriptional activation of NME2 expression is a specific and common feature of chronic myeloid leukemia.
We have previously identified NME2 (Nm23-H2) as a tumor antigen in a patient with chronic myeloid leukemia (CML). Here we investigated the association between NME2 and Bcr-Abl. NME2 protein was highly overexpressed in the cytoplasm of peripheral blood mononuclear cells from 29/30 patients with CML at diagnosis and 10/10 patients resistant to imatinib. Protein was overexpressed in the absence of increased levels of mRNA and was limited to Bcr-Abl + populations, being absent from Bcr-Abl - patient cells, normal donors and 14/15 acute myeloid leukemia (AML) samples. Furthermore, the Bcr-Abl dependent overexpression of NME2 protein was reversed specifically by tyrosine kinase inhibitor (TKI) treatment of Ba/F3 expressing wild-type and TKI-sensitive, but not TKI-resistant, mutants of Bcr-Abl. The post-transcriptional up-regulation of the tumor antigen NME2 is therefore a common and specific property of CML closely associated with Bcr-Abl activity.


Oncosuppressive and oncogenic activity of the sphingolipid-metabolizing enzyme β-galactosylceramidase.
β-galactosylceramidase (GALC) is a lysosomal enzyme that removes β-galactose from β-galactosylceramide, leading to the formation of the oncosuppressor metabolite ceramide. Recent observations have shown that GALC may exert opposite effects on tumor growth by acting as an oncosuppressive or oncogenic enzyme depending on the different experimental approaches, in vitro versus in vivo observations, preclinical versus clinical findings, and tumor type investigated. This review will recapitulate and discuss the contrasting experimental evidence related to the impact of GALC on the biological behavior of cancer and stromal cells and its contribution to tumor progression.


TACE/ADAM-17 enzymatic activity is increased in response to cellular stimulation.
Tumor necrosis factor-alpha converting enzyme (TACE/ADAM-17) is a metalloprotease disintegrin that cleaves a variety of membrane proteins, releasing ("shedding") their extracellular domains from cells. Most TACE-mediated shedding events occur at low basal rates that are enhanced by treatment of cells with a variety of stimuli. To study the mechanism of induced shedding, we developed a peptide-cleavage assay that measures the cellular TACE activity. In unstimulated cells, cleavage of a TNFalpha processing-site peptide was mediated mainly by enzymes other than TACE. However, stimulation of cells with phorbol-12-myristate-13-acetate (PMA) increased peptide cleavage in a TACE-dependent manner. PMA treatment did not increase the amount of TACE on the cell surface. Moreover, the cytoplasmic domain of TACE was not required for the induced activity. Based on these observations, induction of TACE-mediated shedding events occurs at least in part via an increase in the enzymatic activity of cellular TACE, independent of its cytoplasmic domain.


CTRP12 alleviates cardiomyocyte ischemia‑reperfusion injury via regulation of KLF15.
Myocardial ischemia‑reperfusion (I/R) serves a crucial role in myocardial infarction. C1q/TNF‑related protein 12 (CTRP12) is a secretory protein involved in metabolism. It has been reported that CTRP12 participates in the regulation of numerous cardiovascular diseases. However, its role in myocardial I/R injury remains unclear. In the present study, the left anterior descending coronary artery in mice was ligated to establish a mouse I/R model. A myocardial hypoxia‑reoxygenation (H/R) cell model was also established. Cardiomyocyte injury was evaluated using hematoxylin and eosin staining, Cell Counting Kit‑8 and a lactate dehydrogenase (LDH) kit. The expression levels of CTRP12 and Krueppel‑like factor 15 (KLF15) in murine myocardial tissues and H9c2 cells were determined using reverse transcription‑quantitative PCR and western blotting, as KLF15 was previously reported to protect against I/R‑induced cardiomyocyte damage. Furthermore, inflammatory factors TNF‑α, IL‑1β and IL‑6 were analyzed using ELISA while apoptosis was assessed using TUNEL assays and western blotting. Moreover, the activity of the CTRP12 promoter was determined using a dual‑luciferase reporter assay. The results demonstrated that I/R surgery markedly exacerbated myocardial tissue damage, whereas H/R treatment significantly reduced cell viability and significantly increased LDH activity as well as the release of inflammatory factors and apoptosis. I/R and H/R induction significantly reduced the expression levels of CTRP12 and KLF15. CTRP12 overexpression significantly alleviated H/R‑induced cell injury and significantly inhibited inflammation and apoptosis. Further analysis demonstrated that KLF15 could significantly promote the activity of the CTRP12 promoter. However, following CTRP12 knockdown, KLF15 overexpression exacerbated cell injury, inflammation and apoptosis. In conclusion, the present study demonstrated that CTRP12 may mitigate inflammation and apoptosis in H/R‑induced cardiomyocytes, possibly via the regulation of KLF15, which provided a theoretical basis for the potential treatment of I/R‑induced myocardial infarction.


Monitoring of the benzene and toluene contents in human milk.
Twenty-three samples of human milk collected from the milk bank of a children's hospital were analysed with a view to monitoring the possible presence of some of the most common aromatic hydrocarbons (benzene and toluene) and to quantify their concentrations. The analysis was carried out by the "purge and trap" technique combined with gas chromatography and with the use of the mass spectrometer as detector. The hydrocarbons themselves were used in a deuterated form as internal standards. The analysis of the data showed the presence of both hydrocarbons, even though their quantity was much lower than that detected in other foods.


The transformation of the Synaptonemal Complex into the 'elimination chromatin' in Bombyx mori oocytes.
In Bombyx mori oocytes the synaptonemal complexes are retained in modified from pachytene to metaphase I. At the end of pachytene the length and width of the lateral components of the complex increase, whereafter the complexes become compacted during later stages of the meiotic prophase. Ultimately, at metaphase I the modified synaptonemal complexes of individual bivalents fuse to form a more or less continuous sheet between the homologus chromosomes. This sheet corresponds to the structure historically known as the 'elimination chromatin'. It is concluded that in the absence of crossing over and chiasma formation in Bombyx mori females the retainment and subsequent modification of the synaptonemal complex has evolved as a substitute mechanism to ensure regular disjunction of the bivalents.


Molecular docking appraisal of Dysphania ambrosioides phytochemicals as potential inhibitor of a key triple-negative breast cancer driver gene.
Triple-negative breast cancer (TNBC) is a lethal and aggressive breast cancer subtype. It is characterized by the deficient expression of the three main receptors implicated in breast cancers, making it unresponsive to hormone therapy. Hence, an existing need to develop a targeted molecular therapy for TNBC. The PI3K/AKT/mTOR signaling pathway mediates critical cellular processes, including cell proliferation, survival, and angiogenesis. It is activated in approximately 10-21% of TNBCs, emphasizing the importance of this intracellular target in TNBC treatment. AKT is a prominent driver of the PI3K/AKT/mTOR pathway, validating it as a promising therapeutic target. Dysphania ambrosioides is an important ingredient of Nigeria's traditional herbal recipe for cancer treatment. Thus, our present study explores its anticancer properties through a structure-based virtual screening of 25 biologically active compounds domiciled in the plant. Interestingly, our molecular docking study identified several potent inhibitors of AKT 1 and 2 isoforms from D. ambrosioides. However, cynaroside and epicatechin gallate having a binding energy of - 9.9 and - 10.2 kcal/mol for AKT 1 and 2, respectively, demonstrate considerable drug-likeness than the reference drug (capivasertib), whose respective binding strengths for AKT 1 and 2 are - 9.5 and - 8.4 kcal/mol. Lastly, the molecular dynamics simulation experiment showed that the simulated complex systems of the best hits exhibit structural stability throughout the 50 ns run. Together, our computational modeling analysis suggests that these compounds could emerge as efficacious drug candidates in the treatment of TNBC. Nevertheless, further experimental, translational, and clinical research is required to establish an empirical clinical application. A structure-based virtual screening and simulation of Dysphania ambrosioides phytochemicals in the active pocket of AKT 1 and 2 isoforms.


Aqueous Extract of Wood Ear Mushroom, Auricularia polytricha (Agaricomycetes), Demonstrated Antiepileptic Activity against Seizure Induced by Maximal Electroshock and Isoniazid in Experimental Animals.
Auricularia polytricha is a popular mushroom found all over the world. This article describes a study of the antiepileptic effect of A. polytricha, a mushroom that is used traditionally for treating asthma, rheumatism, tumors, cough, fever, and epilepsy, and for its antimicrobial effect. We carried out toxicity studies to identify a standard dose of A. polytricha aqueous extract; maximal electroshock (MES)- and isoniazid (INH)-induced seizures in albino mice were used to screen for the extract's antiepileptic activity. Per Organisation for Economic Co-operation and Development Guideline 423, up to 2000 mg/kg body weight of extract was toxic. Animals were treated with aqueous extract at doses of 200, 400, and 600 mg/kg body weight. Phenytoin was used as the reference anticonvulsant drug for comparison. The investigation found a significant interruption in INH-induced clonic seizure. During MES, we found a reduction in the period of hind leg extensor phase; mice exhibited a significant decrease in the duration of hind limb extension after being treated with 400 and 600 mg/kg doses of A. polytricha. Comparable results were obtained in the INH group, as the extract seemed to delay the onset of a clonic seizure. The aqueous extract of A. polytricha showed antiepileptic action against MES- and INH-induced epilepsy in the mice. This extract, however, requires additional study in order to completely explain its active ingredients and their mechanisms of action.


The Who, What, and Why of Drug Discovery and Development.
Inquiry into declining pharmaceutical R&D efficiency has focussed on 'what' can be improved, with only brief thought given to 'who' can be improved. Here, we argue that enabling people in the idea-to-product chain to have a more holistic knowledge of the behaviours and incentives of each other can optimise R&D.


Decreased survival of C/EBP beta-deficient keratinocytes is due to aberrant regulation of p53 levels and function.
Recent studies have identified roles for C/EBPbeta in cellular survival and tumorigenesis, however, the mechanisms through which C/EBPbeta regulates these processes are not fully understood. Previously, we demonstrated that C/EBPbeta(-/-) mice are resistant to carcinogen-induced skin tumorigenesis and in response to topical carcinogen treatment display a 17-fold increase in keratinocyte apoptosis compared to wild-type mice. Here, we have investigated the mechanisms through which C/EBPbeta regulates apoptosis in response to carcinogenic stress. Analysis of carcinogen-treated C/EBPbeta(-/-) mouse skin revealed a striking increase in the number of p53 immunopositive keratinocytes in the epidermis of C/EBPbeta(-/-) mice compared to wild-type mice and this increase was temporally associated with a concomitant anomalous increase in apoptosis. The increased levels of p53 were functional as Mdm2, Bcl-2, C/EBPalpha and p21 were differentially regulated in the epidermis of carcinogen-treated C/EBPbeta(-/-) mice. The increase in p53 protein was not associated with an increase in p53 mRNA levels. To determine whether p53 is required for the increased apoptosis in C/EBPbeta(-/-) mice, C/EBPbeta/p53 compound knockout mice were generated. Carcinogen-treated C/EBPbeta/p53 compound knockout mice did not display increased apoptosis demonstrating p53 is required for the proapoptotic phenotype in C/EBPbeta(-/-) mice. Our results demonstrate that altered keratinocyte survival in C/EBPbeta(-/-) mice results from aberrant regulation of p53 protein and function and indicate C/EBPbeta has a role in the negative regulation of p53 protein levels in response to carcinogen-induced stress.


Risk factors for violence against women by intimate partners in Sakarya, Turkey.
Intimate partner violence (IPV) is an important public health problem in many countries including Turkey, with serious short- and long-term physical and mental health consequences, as well as significant social and public health costs. To understand the violence characteristics and the individual risk factors for both the married women victims and the perpetrator partners are the objectives of this study. 243 married women, who have consulted to hospitals and village clinics between 2008 and 2009 with the mean age of 36.76 (SD = 11.59), accepted to participate in the study voluntarily. Participants were divided into two groups as "victimized" (58.8%, n = 143) and "non-victimized" (41.2%, n = 100) women. Victimized group consisted of women who have had any type of violence from their husbands during their life. A 61-item questionnaire was prepared that included items on demographics, IPV experience, attitudes about IPV and knowledge of legal rights. As compared to non-victimized group, women in victimized group mostly lived in villages than in town/city centers, were illiterate, had bride-price agreement and were younger when married. Place of accommodation, age and number of pregnancies were associated with the risk of victimization. In order to prevent IPV, further qualitative research is required to evaluate the involved risk factors and training programs on how to behave against violence in association with the legal rights should be designed for poorly educated women in their natural settings and hospitals.


Can Generalist Predators Control Bemisia tabaci?
The whitefly, Bemisia tabaci, has developed resistance to many insecticides, renewing interest in the biological control of this global pest. Generalist predators might contribute to whitefly suppression if they commonly occur in infested fields and generally complement rather than interfere with specialized natural enemies. Here, we review literature from the last 20 years, across US cropping systems, which considers the impacts of generalist predators on B. tabaci. Laboratory feeding trials and molecular gut content analysis suggest that at least 30 different generalist predator species willingly and/or regularly feed on these whiteflies. Nine of these predators appear to be particularly impactful, and a higher abundance of a few of these predator species has been shown to correlate with greater B. tabaci predation in the field. Predator species often occupy complementary feeding niches, which would be expected to strengthen biocontrol, although intraguild predation is also common and might be disruptive. Overall, our review suggests that a bio-diverse community of generalist predators commonly attacks B. tabaci, with the potential to exert substantial control in the field. The key challenge will be to develop reduced-spray plans so that generalist predators, and other more specialized natural enemies, are abundant enough that their biocontrol potential is realized.


Nuclear matrix from resting and concanavalin A-stimulated human lymphocytes.
Concanavalin A-induced transformation and proliferation of human peripheral blood lymphocytes was found to be accompanied by morphological and biochemical changes of the nuclear matrix. Nuclear matrix spheres increased in size as well as in protein and RNA content. Experimental data suggesting the involvement of the nuclear matrix in DNA replication processes are also presented.


Topographical localization in the olivocerebellar projection in the rat: an autoradiographic study.
The topographical organization of the olivocerebellar projection in the rat was examined by making small localized injections of tritiated leucine into the inferior olive (12 cases). The overall projection pattern is displayed in the form of a topographical map, revealing that the olivocerebellar projection is organized in sagittally orientated strips of cerebellar cortex, each of which receives its terminations from a discrete portion of the olive. The projection pattern is compared to that in the cat. In addition, the laterality of the projection is discussed.


Structure of internalin InlK from the human pathogen Listeria monocytogenes.
Listeria monocytogenes is a human pathogen that employs a wide variety of virulence factors in order to adhere to, invade, and replicate within target cells. Internalins play key roles in processes ranging from adhesion to receptor recognition and are thus essential for infection. Recently, InlK, a surface-associated internalin, was shown to be involved in Listeria's ability to escape from autophagy by recruitment of the major vault protein (MVP) to the bacterial surface. Here, we report the structure of InlK, which harbors four domains arranged in the shape of a "bent arm". The structure supports a role for the "elbow" of InlK in partner recognition, as well as of two Ig-like pedestals intercalated by hinge regions in the projection of InlK away from the surface of the bacterium. The unusual fold and flexibility of InlK could be essential for MVP binding and concealment from recognition by molecules involved in the autophagic process.


Uterine artery suture: a preventive approach for pernicious placenta previa.
Pernicious placenta previa is an obstetric complication that results in a crisis situation for many patients. Most existing options deal with the problems after delivery leading to unfavorable outcomes. To circumvent this situation, a few preventive simple surgical options have been discussed in this paper. The highlight of the approach is the application of ascending uterine artery suture that drastically reduced the amount of bleeding as compared to that observed in regular surgeries. Combined with the placenta accrete location, wedge resection and 'patch' homeostasis of the serous membrane, the surgical modifications adopted effectively reduced the need for hysterectomy thus preserving the fertility of the patients for the future. Since fetuses were safely removed within 1-2 min, the Apgar scores of the neonates were also largely unaffected.


A review of the epidemiology of oral and pharyngeal carcinoma: update.
Oral and pharyngeal cancers are the sixth most common cancers internationally. In the United States, there are about 30,000 new cases of oral and pharyngeal cancers diagnosed each year. Furthermore, survival rates for oral and pharyngeal cancers have not significantly improved over the last three decades. This review examines the scientific literature surrounding the epidemiology of oral and pharyngeal cancers, including but not limited to risk factors, disparities, preventative factors, and the epidemiology in countries outside the United States. The literature review revealed that much of the research in this field has been focused on alcohol, tobacco, and their combined effects on oral and pharyngeal cancers. The literature on oral and pharyngeal cancer disparities among racial groups also appears to be growing. However, less literature is available on the influence of dietary factors on these cancers. Finally, effective interventions for the reduction of oral and pharyngeal cancers are discussed.


Defensins: Transcriptional regulation and function beyond antimicrobial activity.
Defensins are one the largest group of antimicrobial peptides and are part of the innate defence. Defensins are produced by animals, plants and fungi. In animals and plants, defensins can be constitutively or differentially expressed both locally or systemically which confer defence before and a stronger response after infection. Immune signalling pathways regulate the gene expression of defensins. These pathways include cellular receptors, which recognise pathogen-associated molecular patterns and are found both in plants and animals. After recognition, signalling pathways and, subsequently, transcriptional factors are activated. There is an increasing number of novel functions in defensins, such as immunomodulators and immune cell attractors. Identification of defensin triggers could help us to elucidate other new functions. The present article reviews the different elicitors of defensins with a main focus on human, fish and marine invertebrate defensins.


Enhanced Versatility in Thorium Removal: Mesoporous Silica-Coated Magnetic Nanoparticles Functionalized by Phenanthroline Diamide as a Selective Adsorbent.
In order to promote the sustainable development of nuclear energy through thorium (Th(IV)) recycling, we synthesized SiO2-coated magnetic functional nanocomposites (SiO2@Fe3O4) that were modified with 2,9-diamide-1,10-phenanthroline (DAPhen) to serve as an adsorbent for Th(IV) removal. SiO2@Fe3O4-DAPhen showed effective Th(IV) adsorption in both weakly and strongly acidic solutions. Owing to its porous structure that facilitated rapid adsorption kinetics, equilibrium was achieved within 5 and 0.5 min at pH 3 and 1 mol L-1 HNO3, respectively. In weakly acidic solutions, Th(IV) primarily formed chemical coordination bonds with DAPhen groups, while in strongly acidic solutions, the dominant interaction was electrostatic attraction. Density functional theory (DFT) calculations indicated that electrostatic attraction was weaker compared to chemical coordination, resulting in reduced diffusion resistance and consequently faster adsorption rates in strongly acidic solutions. Furthermore, SiO2@Fe3O4-DAPhen exhibited a high adsorption capacity for Th(IV); it removed Th(IV) through chelation and electrostatic attraction at pH 3 and 1 mol L-1 HNO3, with maximum adsorption capacities of 833.3 and 1465.7 mg g-1, respectively. SiO2@Fe3O4-DAPhen also demonstrated excellent tolerance to salinity, adsorption selectivity, and radiation resistance, thereby highlighting its practical potential for Th(IV) removal in diverse contaminated water sources. Hence, SiO2@Fe3O4-DAPhen represents a promising choice for the rapid and efficient removal of Th(IV).


Erythropoietin mediates hepcidin expression in hepatocytes through EPOR signaling and regulation of C/EBPalpha.
Hepcidin is the principal iron regulatory hormone, controlling the systemic absorption and remobilization of iron from intracellular stores. Recent in vivo studies have shown that hepcidin is down-regulated by erythropoiesis, anemia, and hypoxia, which meets the need of iron input for erythrocyte production. Erythropoietin (EPO) is the primary signal that triggers erythropoiesis in anemic and hypoxic conditions. Therefore, a direct involvement of EPO in hepcidin regulation can be hypothesized. We report here the regulation of hepcidin expression by EPO, in a dose-dependent manner, in freshly isolated mouse hepatocytes and in the HepG2 human hepatocyte cell model. The effect is mediated through EPOR signaling, since hepcidin mRNA levels are restored by pretreatment with an EPOR-blocking antibody. The transcription factor C/EBPalpha showed a pattern of expression similar to hepcidin, at the mRNA and protein levels, following EPO and anti-EPOR treatments. Chromatin immunoprecipitation experiments showed a significant decrease of C/EBPalpha binding to the hepcidin promoter after EPO supplementation, suggesting the involvement of this transcription factor in the transcriptional response of hepcidin to EPO.


Pharmacogenes that demonstrate high association evidence according to CPIC, DPWG, and PharmGKB.
Different levels of evidence related to the variable responses of individuals to drug treatment have been reported in various pharmacogenomic (PGx) databases. Identification of gene-drug pairs with strong association evidence can be helpful in prioritizing the implementation of PGx guidelines and focusing on a gene panel. This study aimed to determine the pharmacogenes with the highest evidence-based association and to indicate their involvement in drug-gene interactions. The publicly available datasets CPIC, DPWG, and PharmGKB were selected to determine the pharmacogenes with the highest drug outcome associations. The upper two levels of evidence rated by the three scoring methods were specified (levels A-B in CPIC, 3-4 in DPWG, or 1-2 levels in PharmGKB). The identified pharmacogenes were further ranked in this study based on the number of medications they interacted with. Fifty pharmacogenes, with high to moderately high evidence of associations with drug response alterations, with potential influence on the therapeutic and/or toxicity outcomes of 152 drugs were identified. CYP2D6, CYP2C9, CYP2C19, G6PD, HLA-B, SLCO1B1, CACNA1S, RYR1, MT-RNR1, and IFNL4 are the top 10 pharmacogenes, where each is predicted to impact patients' responses to ≥5 drugs. This study identified the most important pharmacogenes based on the highest-ranked association evidence and their frequency of involvement in affecting multiple drugs. The obtained data is useful for customizing a gene panel for PGx testing. Identifying the strength of scientific evidence supporting drug-gene interactions aids drug prescribers in making the best clinical decision.


Using physicochemical and biological parameters for the evaluation of water quality and environmental conditions in international wetlands on the southern part of Lake Urmia, Iran.
The Kani Barazan and Yadegarlou wetlands in the southern part of Lake Urmia (Iran) have been substantially modified due to human activities and anthropogenic use. In recent years, freshwater-based eco-biological studies to recognize the quality of water resources have been greatly expanded. Microalgae and Cyanophyta are considered important bioindicators for the evaluation of water quality and wetland health worldwide. Herein, 22 microalgae and 5 Cyanophyta genera were identified in both wetlands, in which Cyanophyta has mainly caused blooms. Principal components analysis (PCA) was carried out based on links between the distribution of microalgae and Cyanophyta with physical and chemical parameters. The data showed that depth, turbidity, and the temperature had a significant influence on the microalga and Cyanophyta communities in both wetlands. Based on the biological properties, it seems that the Kani Barazan and Yadegarlou international wetlands experience meso-eutrophic conditions. The integration of the physical, chemical and biological parameters with the water quality index (WQI) revealed that both wetlands were polluted as a consequence of human activities. Moreover, a close relationship between WQI and the biological parameters was documented. Thus, we concluded that microalgae and Cyanophyta communities, their abundance patterns, and water quality changes could provide valuable data for the conservation of the Kani Barazan and Yadegarlou international wetlands.


Magnetic resonance imaging of midline pediatric cerebral neoplasms.
Over 80 cases of pediatric cerebral disease including 60 brain tumors, have been studied by low field 0.12 Tesla resistive NMR proton imaging. In addition, experience has been gained with proton imaging on a superconducting high field 1.5 Tesla system. Comparison of NMR images at both field strength, and comparison of the images to those obtained with high resolution CT is made. Examples of how NMR images can help to plan patient management is presented.


Implementing substance abuse group therapy clinical trials in real-world settings: challenges and strategies for participant recruitment and therapist training in the Women's Recovery Group Study.
Open-enrollment group therapy research is challenged by the participant recruitment necessary to ensure continuous group enrollment. We present successful strategies to overcome the following barriers during the Women's Recovery Group (WRG) two-site clinical trial (N = 158): maintenance of sample size and balanced gender randomization during continuous enrollment, maintenance of group attendance, and training and retention of therapists over the 24-month continuous group enrollment. To increase recruitment, we targeted referral sources yielding the highest enrollment conversion at each site. Group sessions were consistently held regardless of group size. Therapists were trained in two teams allowing for coverage and uninterrupted treatment over 24 months. At both sites recruitment and enrollment increased with each successive quarter. Sample size and end date targets were met without disruptions in treatment. Group therapists reported high satisfaction with their training and treatment experiences. These strategies supported targeted enrollment and study duration, stability of open-enrollment group therapy frame, and therapist retention and satisfaction. Applying these strategies can aid in providing evidence-based group therapy in both clinical and research settings.


Survival predictors of metastatic angiosarcomas: a surveillance, epidemiology, and end results program population-based retrospective study.
Angiosarcomas (AS) have poor prognosis and often metastasize to distant sites. The potential predictors of metastatic angiosarcomas (MAS) have not been extensively investigated. The main objective of this study was to identify survival predictors of MAS. Surveillance, Epidemiology, and End Results (SEER) datasets were used to identify patients with MAS from 2010 to 2016. Risk predictors were determined with the aid of Kaplan-Meier and Cox regression model analyses. A total of 284 MAS patients met the study entry criteria. Among these, 121 patients (42.6%) were diagnosed with metastasis in bone, 26 in brain (9.2%), 86 in liver (30.3%) and 171 in lung (60.2%). Overall, 96 patients (33.8%) had two or more metastatic sites. The 1- and 3-year overall survival (OS) rates were 20.8 and 3.8% while 1- and 3-year cancer-specific survival (CSS) rates were 22.0 and 5.2%, respectively. Cox regression analysis revealed chemotherapy, radiation treatment (RT) and tumor size ≤10 cm as independent favorable predictors of OS. In terms of CSS, tumor grade IV, tumor size > 10 cm and absence of chemotherapy were independent adverse predictors. Surgery did not prolong survival outcomes (both OS and CSS) in the current cohort. MAS is associated with extremely poor survival. Chemotherapy, RT, and tumor size are independent predictors of OS. Chemotherapy and tumor size are independent prognostic factors of CSS. Chemotherapy is therefore recommended as the preferred treatment option for MAS patients.


Progress on nanostructured electrochemical sensors and their recognition elements for detection of mycotoxins: A review.
Nanomaterial-embedded sensors have been developed and applied to monitor various targets. Mycotoxins are fungal secondary metabolites that can exert carcinogenic, mutagenic, teratogenic, immunotoxic, and estrogenic effects on humans and animals. Consequently, the need for the proper regulation on foodstuff and feed materials has been recognized from times long past. This review provides an overview of recent developments in electrochemical sensors and biosensors employed for the detection of mycotoxins. Basic aspects of the toxicity of mycotoxins and the implications of their detection are comprehensively discussed. Furthermore, the development of different molecular recognition elements and nanomaterials required for the detection of mycotoxins (such as portable biosensing systems for point-of-care analysis) is described. The current capabilities, limitations, and future challenges in mycotoxin detection and analysis are also addressed.


The effects of temperature on the respiration and production of the freshwater nematode Anonchus sp.
Growth and respiration were measured in a species of Anonchus (Nematoda: Plectidae) at 5°C, 10°C, 15°C, 20°C and 25°C. At 5°C no growth was measurable but the organisms remained active. Maximum production occurred at 15°C, but the highest rate of growth occurred at 20°C. Thus, adult size attained is dependent on the temperature of growth. Respiratory energy losses derived from Cartesian diver microrespirometry, increased with temperature up to 25°C. Regression coefficients (b values) derived from a log log linear regression of weight against oxygen consumption varied between 0.574-1.793, the lowest value being attained at 5°C, the highest at 20°C. Based on Q10, production and respiratory energy losses the optimum temperatures for Anonchus appears to lie between 10°C-15°C.


N-methyl-D-aspartate and kainate receptor gene expression in hippocampal pyramidal and granular neurons in the kindling model of epileptogenesis.
To investigate the changes underlying kindling epileptogenesis in the rat hippocampus, the levels of the messenger RNAs encoding for the subunits of the N-methyl-D-aspartate-receptor (1, 2A-D) and the kainate-receptor (1, -2, GluR-5, -6, -7) were determined in hippocampal principal neurons using in situ hybridization techniques and semi-quantitative analysis of the autoradiograms. Schaffer collateral-commissural pathway kindled rats were investigated at three different stages of kindling acquisition, always 24 h after the last stimulation. Furthermore, fully kindled rats were studied at long-term (28 days) after termination of kindling stimulations. NR1 messenger RNA levels were slightly decreased in CA1 area of fully kindled animals. In the fascia dentata region, a minor increase of NR2A and NR2B transcripts was found at all stages of kindling acquisition. KA-2 messenger RNA was enhanced in all hippocampal subfields during kindling development. However, none of these changes persisted at long-term after the last seizure and only the low-abundant GluR-7 expression was slightly depressed in the fascia dentata. From our observations we conclude that it is unlikely that alterations in N-methyl-Daspartate or kainate receptor gene expression play an important role in kindling acquisition or maintenance.


Colorimetric immunosensing using liposome encapsulated MnO2 nanozymes for SARS-CoV-2 antigen detection.
Development of specific signal reporters with signal amplification effect are highly needed for sensitive and accurate detection of pathogen. Herein, we design a colorimetric immunosensing nanosystem based on liposome encapsulated quantum dots-sized MnO2 nanozyme (MnO2QDs@Lip) as a signal reporter for ultrasensitive and fast detection of SARS-CoV-2 antigen. The pathogenic antigens captured and separated by antibody-conjugated magnetic beads (MBs) are further connected with antibody-modified MnO2QDs@Lip to form a sandwich-like immunocomplex structure. After triggered release, MnO2 QDs efficiently catalyze colorless 3,3',5,5'-tetramethylbenzidine (TMB) to blue oxidized TMB, which can be qualitatively observed by naked eyes and quantitatively analyzed by UV-Vis spectra or smartphone platforms. By taking advantages of immuno-magnetic separation, excellent peroxidase-like catalytic activity of MnO2 QDs, and high encapsulation efficiency of MnO2QDs@Lip, ultrasensitive detection of SARS-CoV-2 antigen ranging from 0.1 pg/mL to 100 ng/mL is achieved within 20 min. The limit of detection (LOD) is calculated to be 65 fg/mL in PBS buffer. Furthermore, real clinical samples of SARS-CoV-2 antigens can be effectively identified by this immunosensing nanosystem with excellent accuracy. This proposed detection nanosystem provides a strategy for simple, rapid and ultrasensitive detection of pathogens and may shed light on the development of new POCT detection platforms for early diagnosis of pathogens and surveillance in public health.


Transition metal spin state energetics and noninnocent systems: challenges for DFT in the bioinorganic arena.
Although density functional theory (DFT) provides a generally good description of transition metal systems, we have identified several cases, involving Fe(III) porphyrins and related systems, where common functionals fail to correctly describe the energetics of the different low-lying spin states. The question of metal- versus ligand-centered oxidation in high-valent transition metal complexes is also a challenging one for DFT calculations, as I have tried to illustrate with examples from among porphyrin, corrole, biliverdine, and NO complexes. In a number of cases, I have compared results obtained with different exchange-correlation functionals; in addition, I have added a discussion on the relative performance of pure versus hybrid functionals. Finally, I have offered some thoughts on the role that traditional wavefunction-based ab initio methods, now essentially absent from the bioinorganic arena, might play in the future.


Neurokinin-1 Receptor (NK-1R) Antagonists as a New Strategy to Overcome Cancer Resistance.
Nowadays, the identification of new therapeutic targets that allow for the development of treatments, which as monotherapy, or in combination with other existing treatments can contribute to improve response rates, prognosis and survival of oncologic patients, is a priority to optimize healthcare within sustainable health systems. Recent studies have demonstrated the role of Substance P (SP) and its preferred receptor, Neurokinin 1 Receptor (NK-1R), in human cancer and the potential antitumor activity of NK-1R antagonists as an anticancer treatment. In this review, we outline the relevant studies published to date regarding the SP/NK-1R complex as a key player in human cancer and also evaluate if the repurposing of already marketed NK-1R antagonists may be useful in the development of new treatment strategies to overcome cancer resistance.


Glucose starvation is required for insulin stimulation of glucose uptake and metabolism in cultured microvascular endothelial cells.
In the present study we determined the uptake and disposition of glucose in serum-deprived rabbit coronary microvessel endothelial (RCME) cells. RCME cells exhibited stereospecific hexose uptake inhibited by cytochalasin B. Pretreatment of the RCME cells with potassium cyanide or 2,4-dinitrophenol inhibited 2-deoxyglucose uptake but not 3-O-methylglucose transport. A major proportion (30-60%) of the 2-deoxyglucose present in the RCME cells was not phosphorylated. These two observations suggested that the rate-limiting step in the uptake of 2-deoxyglucose was not transport but rather the phosphorylation of 2-deoxyglucose to 2-deoxyglucose 6-phosphate. When glucose-deprived cells were incubated 2 hr with [U-14C]glucose the disposition of the label was as follows: glycogen 60%, acid-soluble fraction 30%, and lipid less than 5%. In contrast glucose-fed cells exhibited lower overall glucose incorporation, and a slightly different disposition: glycogen 45%, acid-soluble fraction 50%, and lipid 5%. Glucose-deprived RCME cells also exhibited greater basal levels of 2-deoxyglucose uptake compared to glucose-fed cells. RCME cells incubated in the absence of glucose and serum for 16 hr exhibited dose-dependent insulin stimulation of hexose uptake and subsequent metabolism to macromolecules (i.e., glycogen and the acid-soluble fraction). Significant effects of insulin were observed with concentrations as low as 2 x 10(-10) M, well within the physiological range. In contrast, cells preincubated in serum-free culture medium containing 5.5 mM glucose did not exhibit insulin-enhanced hexose uptake or glucose metabolism (even at doses as high as 10(-7) M). These studies indicate that the effects of insulin on rabbit coronary microvascular endothelial cell glucose uptake and metabolism require both serum and glucose deprivation.


Long-term oral Candida colonization, mucositis and salivary function after head and neck radiotherapy.
The aim of this study was to follow the long-term effects of radiation therapy of head and neck malignancies on oral yeast colonization, mucositis and salivary function. Included in this prospective study were 32 patients with intended radiation therapy of a malignancy of the head and neck. In all patients the salivary glands lay within the radiation field and the patients had at least five teeth. The first examination was performed after oral hygiene instruction and removal of questionable teeth before the start of radiotherapy. The following examinations were conducted after 3, 6, 9 and 12 months. Together with the quantitative determination of Candida colonization, three "mucositis" variables were assessed: (1) examiner-rated mucositis score (LENT/SOMA), (2) patient-rated mucositis symptoms, and (3) scintigraphic salivary excretion fraction. The maximum Candida colonization was found 6 months after radiation therapy and this declined to above normal values after 12 months. Salivary flow was at a minimum 6 months after radiation therapy and had slightly recovered by 12 months. Examiner-rated mucositis and patient-rated xerostomia showed no significant recovery after 6 or 12 months. The results of this study show slight recovery of the oral ecological system. Although the causal role of a single parameter is not clear, persistently elevated Candida colonization should be taken into account therapeutically.


A robust qualitative transcriptional signature for the correct pathological diagnosis of gastric cancer.
Currently, pathological examination of gastroscopy biopsy specimens is the gold standard for gastric cancer (GC) diagnosis. However, it has a false-negative rate of 10-20% due to inaccurate sampling locations and/or insufficient sampling amount. A signature should be developed to aid the early diagnosis of GC using biopsy specimens even when they are sampled from inaccurate locations. We extracted a robust qualitative transcriptional signature, based on the within-sample relative expression orderings (REOs) of gene pairs, to discriminate both GC tissues and adjacent-normal tissues from non-GC gastritis, intestinal metaplasia and normal gastric tissues. A signature consisting of two gene pairs for GC diagnosis was identified and validated in data of both biopsy specimens and surgical resection specimens pooled from publicly available datasets measured by different laboratories with different platforms. For gastroscopy biopsy specimens, 96.20% of 79 non-GC tissues were correctly identified as non-GC, and 96.84% of 158 GC tissues and six of seven adjacent-normal tissues were correctly identified as GC. For surgical resection specimens, 98.37% of 2560 GC tissues and 97.28% of 221 adjacent-normal tissues were correctly identified as GC. Especially, 97.67% of the 257 GC patients at stage I were exactly diagnosed as GC. We additionally measured 21 GC tissues from seven different GC patients, each with three specimens sampled from three tumor locations with different proportions of the tumor epithelial cell. All these GC tissues were correctly identified as GC, even when the proportion of the tumor epithelial cell was as low as 14%. The qualitative transcriptional signature can distinguish both GC and adjacent-normal tissues from normal, gastritis and intestinal metaplasia tissues of non-GC patients even using inaccurately sampled biopsy specimens, which can be applied robustly at the individual level to aid the early GC diagnosis.


Vanadyl-induced Fenton-like reaction in RNA. An ESR and spin trapping study.
Vanadyl (VO2+) complexed to RNA reacts with hydrogen peroxide in a Fenton-like manner producing hydroxyl radicals (.OH). The hydroxyl radicals can be spin trapped with 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) forming the DMPO-OH spin adduct. In addition, in the presence of ethanol the formation of the hydroxyethyl radical adduct of DMPO (DMPO-ETOH) confirms the production of hydroxyl radicals by the RNA/VO2+ complex. When the reaction between the RNA/VO2+ complex and H2O2 is carried out in the presence of the spin trap 2-methyl-2-nitrosopropane (MNP), radicals produced in the reaction of .OH with RNA are trapped. Base hydrolysis of the MNP-RNA adducts (pH 12) followed by a reduction in the pH to pH 7 after hydrolysis is complete, yields an MNP adduct with a well-resolved ESR spectrum identical to the ESR spectrum obtained from analogous experiments with poly U. The ESR spectrum consists of a triplet of sextets (aN = 1.48 mT, a beta N = 0.25 mT and a beta H = 0.14 mT), indicating that the unpaired nitroxide electron interacts with the nuclei of a beta-nitrogen and beta-hydrogen. The results suggest that the .OH generated in the RNA/VO2+ reaction with H2O2 add to the C(5) carbon of uracil forming a C(6) carbon centered radical. This radical is subsequently spin trapped by MNP.


[Effect of increasing the heat content of the body on the temperature of the skin surface and various regions of the body underling it].
Artificial heating of the body (127.3 +/- 6.9 cal/kg) increased cutaneous temperature and subcutaneous one on the nose by 0.38 +/- 0.09 degrees and 0.38 +/- 0.10 degrees C, resp.; on the back - by 0.16 +/- 0.05 degrees and 0.18 +/- 0.05 degrees C, resp.; on the abdomen - by 0.19 +/- 0.04 degrees and 0.21 +/- 0.03 degrees C, resp.


Norepinephrine induces hepatic fibrogenesis in leptin deficient ob/ob mice.
Leptin's actions on certain cells require a leptin-inducible neurotransmitter, norepinephrine (NE). NE modulates hepatic fibrosis. Therefore, decreased NE may explain why leptin deficiency inhibits hepatic fibrosis. We manipulated adrenergic activity in leptin-deficient ob/ob mice, leptin-sufficient, dopamine beta-hydroxylase deficient (Dbh(-/-)) mice, and HSC cultures to determine if leptin requires NE to activate HSC and induce hepatic fibrosis. ob/ob mice have chronic liver injury, but reduced numbers of HSC. Supplemental leptin increases HSC, suggesting that leptin-dependent, injury-related factors permit expansion of HSC populations. NE also increases HSC numbers and activation, normalizing fibrogenesis. When fed hepatotoxic diets, NE-deficient Dbh(-/-) mice fail to accumulate activated HSC and have impaired fibrogenesis unless treated with adrenergic agonists. NE acts directly on HSC to modulate leptin's actions because leptin increases HSC proliferation and prazosin, an alpha-adrenoceptor antagonist, inhibits this. Thus, leptin permits injury-related increases in adrenergic activity and requires NE to activate HSC and induce hepatic fibrogenesis.


Nucleotide sequence of the chloramphenicol acetyltransferase gene of Streptomyces acrimycini.
The nucleotide sequence of a gene (cat) encoding chloramphenicol acetyltransferase (CAT) in Streptomyces acrimycini was determined. The predicted amino acid sequence demonstrates extensive homology with those of CATs isolated from Gram-negative Enterobacteria, notably with the type III variant encoded by the IncK plasmid R387. Transcript mapping indicates a single cat mRNA with a 5' end coinciding with the AUG codon used for translational initiation in vivo. We also determined the extent of a spontaneous deletion in the 5'-noncoding DNA, which occurs when the gene is cloned in the BamHI site of pBR322 in a specific orientation and which results in constitutive cat expression in Escherichia coli from the tet promoter.


Potential COVID-19 therapeutic approaches targeting angiotensin-converting enzyme 2; An updated review.
COVID-19 has spread swiftly throughout the world posing a global health emergency. The significant numbers of deaths attributed to this pandemic have researchers battling to understand this new, dangerous virus. Researchers are looking to find possible treatment regimens and develop effective therapies. This study aims to provide an overview of published scientific information on potential treatments, emphasizing angiotensin-converting enzyme II (ACE2) inhibitors as one of the most important drug targets. SARS-CoV-2 receptor-binding domain (RBD); as a viral attachment or entry inhibitor against SARS-CoV-2, human recombinant soluble ACE2; as a genetically modified soluble form of ACE2 to compete with membrane-bound ACE2, and microRNAs (miRNAs); as a negative regulator of the expression of ACE2/TMPRSS2 to inhibit SARS-CoV2 entry into cells, are the potential therapeutic approaches discussed thoroughly in this article. This review provides the groundwork for the ongoing development of therapeutic agents and effective treatments against SARS-COV-2.


Effects of atorvastatin in combination with celecoxib and tipifarnib on proliferation and apoptosis in pancreatic cancer sphere-forming cells.
Cancer stem cell (CSC) plays an important role in pancreatic cancer pathogenesis and treatment failure. CSCs are characterized by their ability to form tumor spheres in serum-free medium and expression of CSC related markers. In the present study, we investigated the effect atorvastatin, celecoxib and tipifarnib in combination on proliferation and apoptosis in Panc-1 sphere-forming cells. The sphere-forming cells were isolated from Panc-1 cells by sphere-forming method. These sphere-forming cells showed CSC properties. The levels of CD44, CD133 and ALDH1A1 in the sphere-forming cells were increased. Moreover, Panc-1 sphere-forming cells were resistant to chemotherapeutic drug gemcitabine. Combined atorvastatin with celecoxib and tipifarnib synergistically decreased the sphere forming ability of Panc-1 cells and the drug combination also strongly inhibited cell proliferation and promoted apoptosis in the sphere-forming cells. The effects of the drug combination on the Panc-1 sphere-forming cells were associated with decreases in the levels of CD44, CD133 and ALDH1A1, and suppression of Akt and NF-κB activation. Results of the present study indicate that the combination of atorvastatin, celecoxib and tipifarnib may represent an effective approach for inhibiting pancreatic CSCs.


Proteome alterations in pancreatic ductal adenocarcinoma.
Proteins are the essential functional biomolecules profoundly implicated in all aspects of pancreatic tumorigenesis and its progression. While common genomic factors, such as KRAS, TP53, SMAD4, and CDKN2A have been well recognized in association of pancreatic ductal adenocarcinoma (PDAC), our understanding of functional changes at the proteome level merits further investigation. Malignance associated proteome alterations can be attributed to the convoluted outcomes from genetic, epigenetic and environmental factors in initiating and progressing PDAC, and may reflect on changes in protein expressional level, structure, localization, as well as post-translational modifications (PTMs) status. The study of localized or systemic proteome alterations in PDAC, as well as its precursor lesions, such as pancreatic intraepithelial neoplasia (PanIN) and mucinous pancreatic cystic neoplasm, would provide unique perspectives in elucidating functional molecular events underlying PDAC. While efforts have been made, challenges still exist to comprehensively integrate much of the proteomic discovery to the perspectives gained from genomic studies in the context of biomarker discovery. Novel approaches and data from well-defined longitudinal clinical studies and experimental models are needed to facilitate the study of PDAC and precursor lesions for early detection and intervention.


Association of severe calcium lithogenic activity and bone remodeling markers.
To establish cutoff points for markers of bone remodeling that allow for screening of patients at risk for serious lithogenic activity. We conducted a cross-sectional study with 182 patients (aged between 25 and 60 years) divided into 3 groups: group 1, 56 patients without lithiasis; group 2, 67 patients with light calcium lithiasis; and group 3, 59 patients with severe calcium lithiasis. The criteria for inclusion in and exclusion from the study were established, and light and severe lithogenic activity were defined. Metabolic variables in blood and urine, along with bone densitometry, were studied for the groups. Statistical analysis of the results and preparation of receiver operating characteristic curves to establish optimal cutoff points were performed. The patients in group 3 showed the greatest bone mineral density loss and the highest values for markers of bone remodeling, together with increased 24-hour calciuria. Using the receiver operating characteristic curves developed and based on statistical significance (P = .0001), the following cutoff points for severe lithogenic activity, with a sensitivity between 75% and 85%, were established: β-crosslaps >0.331 ng/mL; osteocalcin >13.2 ng/mL; β-crosslaps/osteocalcin >0.024; 24-hour calciuria >306.6 mg; and fasting urine calcium/creatinine >0.105. Patients with calcium lithiasis and elevated values for osteocalcin, β-crosslaps, β-crosslaps/osteocalcin, 24-hour calciuria, and fasting urine calcium/creatinine may present a high risk of severe lithogenic activity.


Combined effects of high-fat diet and polystyrene microplastic exposure on microplastic bioaccumulation and lipid metabolism in zebrafish.
Extensive use of microplastics (MPs) threatens the safety of aquatic environments and hydrobionts. Increasing the weight of economic fish through high-fat diet (HFD) to increase production is common in aquaculture. However, little is known about the combined effects of MPs and HFD in fish. The aim of this study was to investigate the relationship between adiposity and MP bioaccumulation in fish. Using zebrafish as a vertebrate model, the content of polystyrene (PS) MPs in zebrafish tissues exposed to 5 and 50 μm of 1000 μg/L PS MPs was detected via confocal Raman spectroscopy in normal diet (ND) and HFD. The content of PS MPs in HFD group was significantly higher than that in ND group. The levels of hepatic lipids were significantly elevated in zebrafish subjected to HFD treatment, and this effect was aggravated by exposure to 5 μm PS MPs, and even caused liver injury. Transcriptomic analysis revealed that exposure to PS MPs interferes with hepatic lipid metabolism and energy homeostasis in zebrafish. These results suggests that in addition to controlling the use and performing proper recycling of plastic products in our daily life, we should not blindly increase the weight of fish through HFD. This aids protect the quality of economic fish and prevent MPs from being consumed by humans through the food chain. This study explored the interaction between fish feed culture and environmental pollutants to provide important reference for fish culture.


Albumin use guidelines and outcome in a surgical intensive care unit.
Restrictive albumin use guidelines in the surgical intensive care unit (SICU) will not increase mortality and will result in cost savings. Prospective cohort study. Tertiary teaching hospital. All patients admitted to the SICU from July 1, 2004, through July 1, 2005, were included in this study. Patients in the first 3 quarters of the study were treated with no restriction on albumin use. An organized educational program was initiated by the new intensivist-led critical care team and directed toward the residents, nursing staff, and primary surgical teams. Appropriate albumin use guidelines were instituted in the last quarter. Prospective clinical and outcome data were collected. Albumin use data and costs were obtained from the pharmacy prospective database. A total of 1361 patients were included in the study. A statistically significant reduction in albumin use (54%) was found in the fourth quarter (P = .004), and a substantial cost saving was realized (56% reduction in cost) with the albumin use guidelines. Restrictive use of albumin had no negative impact on ICU mortality. Mean Acute Physiology and Chronic Health Evaluation III scores on ICU day 1 were not different. No significant difference in mean ICU length of stay was noted. Maintained reduction in the use of albumin was documented during the next 6 quarters. The implementation of albumin use guidelines during critical care resuscitation using an educational approach in a SICU is associated with reduced albumin use, significant cost savings, and no negative impact on ICU outcome. Continued educational efforts promoting evidence-based practices in the ICU are warranted.


Female Mice Lacking Estrogen Receptor-α in Hypothalamic Proopiomelanocortin (POMC) Neurons Display Enhanced Estrogenic Response on Cortical Bone Mass.
Estrogens are important regulators of bone mass and their effects are mainly mediated via estrogen receptor (ER)α. Central ERα exerts an inhibitory role on bone mass. ERα is highly expressed in the arcuate (ARC) and the ventromedial (VMN) nuclei in the hypothalamus. To test whether ERα in proopiomelanocortin (POMC) neurons, located in ARC, is involved in the regulation of bone mass, we used mice lacking ERα expression specifically in POMC neurons (POMC-ERα(-/-)). Female POMC-ERα(-/-) and control mice were ovariectomized (OVX) and treated with vehicle or estradiol (0.5 μg/d) for 6 weeks. As expected, estradiol treatment increased the cortical bone thickness in femur, the cortical bone mechanical strength in tibia and the trabecular bone volume fraction in both femur and vertebrae in OVX control mice. Importantly, the estrogenic responses were substantially increased in OVX POMC-ERα(-/-) mice compared with the estrogenic responses in OVX control mice for cortical bone thickness (+126 ± 34%, P < .01) and mechanical strength (+193 ± 38%, P < .01). To test whether ERα in VMN is involved in the regulation of bone mass, ERα was silenced using an adeno-associated viral vector. Silencing of ERα in hypothalamic VMN resulted in unchanged bone mass. In conclusion, mice lacking ERα in POMC neurons display enhanced estrogenic response on cortical bone mass and mechanical strength. We propose that the balance between inhibitory effects of central ERα activity in hypothalamic POMC neurons in ARC and stimulatory peripheral ERα-mediated effects in bone determines cortical bone mass in female mice.


Intraluminal increase of superoxide anion following transient focal cerebral ischemia in rats.
Using a modification of Karnovsky's Mn2+/diaminobenzizine (DAB) technique, we examined the production of superoxide anion (.O-2) in the vascular lumen following transient occlusion and reperfusion of the left middle cerebral artery (MCA) in Sprague-Dawley rats. The MCA was occluded for 2 h using an intraluminal suture method. Zero, 15, 30, and 60 min after reperfusion, animals were perfused transcardially with buffer containing Mn2+ and DAB, and brain samples were prepared for light and electron microscopic examination. The amber reaction deposits of.O-2 were observable to the naked eye along the major cerebral vessels of the ischemic hemisphere after each reperfusion period. Upon microscopic examination the deposits were revealed to be within arterial, capillary, and venular lumen. The amount of reaction deposits in the ischemic hemisphere corresponded to the duration of reperfusion. The formation of.O-2 was suppressed when the perfusate contained superoxide dismutase and when either Mn2+ or DAB was omitted, confirming that the reaction products produced are due to the enhanced production of.O-2. These results show that there is a progressive increase in intraluminal. O-2 during reperfusion following an ischemic insult which may participate in the aggravation of cerebral damage.


Treatment results of radiotherapy for malignant lymphoma of the orbit and histopathologic review according to the WHO classification.
To analyze the results of radiotherapy (RT) for malignant lymphoma of the orbit and to evaluate them compared with the World Health Organization (WHO) classification published in 2001. The data from 29 patients with malignant lymphoma of the orbit treated with RT at Gunma University Hospital between 1978 and 2001 were retrospectively analyzed. Pathologic slides from 23 cases were available and were reviewed by a hematopathologist according to the WHO classification. The original and reviewed diagnoses, patient characteristics, treatment results, and complications were analyzed. In principle, patients with low-grade or indolent lymphoma were treated with RT alone, using 30 Gy as the tumor dose. Survival data were calculated using the Kaplan-Meier method. One case that proved to be a pseudotumor was excluded from evaluation. Of the 28 cases, 25 were Stage IAE, 1 was Stage IIAE, and 2 were Stage IVAE. The median follow-up was 71 months. According to the original classification and the Working Formulation, the 5- and 10-year overall survival rate of patients with low-grade lymphoma was 94% and 73%, respectively. The corresponding rates for those with intermediate-grade lymphoma were 67% and 67% (p = 0.15). In contrast, the WHO classification showed a significant difference in the survival curves. The 5- and 10-year overall survival rate of patients with mucosa-associated lymphoid tissue (MALT) lymphoma was 100% and 88%, respectively; for diffuse large B-cell patients, the rates were both 0% (p < 0.001). In patients with MALT lymphoma, one local and four distant relapses developed; two of them >10 years after initial treatment. All of the relapsed MALT lymphomas were controlled by salvage therapy. Excellent local control and survival can be achieved for patients with orbital MALT lymphoma using RT alone. A precise histopathologic diagnosis using the WHO classification and long-term follow-up for >10 years is recommended.


Diseases associated with multiple sclerosis and epilepsy. A population cohort study of 159,200 middle-aged, urban, native Swedes observed over 10 years (1970-79).
A defined general population of 159,200 male and female native Swedes born in the period 1911-1940 from an urban catchment area of the then only general hospital, was followed over a decade (1970-79) with regard to in-patient hospitalization for all kinds of diagnoses. As a part of this population cohort study, multiple sclerosis cases (n = 351) and epilepsy cases (n = 648) were studied for association with other diseases. Unexpectedly, a cluster of diseases encompassing tuberculosis, bronchial asthma, diabetes mellitus and myocardial infarction, among the diseases associated with multiple sclerosis, also forms a gradient; this suggests a quantitative rather than a qualitative multifactorial model of disease for the understanding of the pathogenesis of MS. In epilepsy, heterogeneity was suggested as being mainly linked to the presence or absence of co-existing alcoholism. Brain tumours in cases of epilepsy were found almost exclusively in the latter subset and prevailing among younger people independent of sex (with an almost 100-fold excess rate of that disease combination as expected by chance only).


Social defeat increases alcohol preference of C57BL/10 strain mice; effect prevented by a CCKB antagonist.
In humans, social stress over long and short term can increase alcohol consumption, but the mechanisms involved are not understood. This study was conducted to examine the effects of social defeat, using the resident/intruder paradigm, on the alcohol preference of "low alcohol drinking" individuals in a colony of C57BL/10 strain mice and the effects of two anxiolytic drugs. Alcohol preference, in a two-bottle choice (8% v/v alcohol or water), was measured, in separate experiments, after either a single experience of social defeat by a resident male mouse, five consecutive daily defeat experiences or one experience per week for 4 weeks. Comparison was made with effects of repeated social defeat on the preference for dilute sucrose. In addition, the actions of the CCKB receptor antagonist, CAM1028, and of diazepam were examined on the effects of repeated defeat experiences. Five consecutive daily defeat experiences had a slow onset effect in increasing alcohol preference and consumption, compared with five daily exposures to a novel environment. A single defeat, or one defeat per week, did not significantly alter alcohol preference or intake. There were no effects of five daily defeat experiences on sucrose preference or consumption. The effect of repeated defeats on alcohol preference was significantly decreased by administration of the CCKB receptor antagonist, CAM1028, prior to each experience, but not by corresponding administration of diazepam. The results show that social stress increases alcohol intake in low alcohol preference C57BL/10 mice and suggest that CCK transmission may be involved in this effect.


Enzyme leakage, trypsinogen activation, and inflammatory response in endoscopic retrograde cholangiopancreatography-induced pancreatitis.
Endoscopic retrograde cholangiopancreatography (ERCP)-induced pancreatitis (EIP) provides an opportunity to study different pathophysiologic events early in the course of acute pancreatitis. To investigate whether the leakage of pancreatic proenzymes (anionic trypsinogen), pancreatic protease activation (carboxypeptidase B activation peptide), cytokine response (interleukin [IL]-1 receptor antagonist, IL-6, and soluble tumor necrosis factor receptor-I) and neutrophil activation (neutrophil gelatinase-associated lipocalin and polymorphonuclear elastase) differ between patients with and without EIP. A second aim was to clarify the temporal relation between these different events. Ninety-nine nonconsecutive patients undergoing ERCP were investigated in the study. Fourteen of 99 patients undergoing ERCP developed mild EIP. Six hours after the investigation the concentration of anionic trypsinogen was significantly higher in patients with EIP than in patients without EIP. The day after ERCP, higher concentrations of anionic trypsinogen, carboxypeptidase B activation peptide, IL-6, and polymorphonuclear elastase were recorded in the EIP group. No significant differences in IL-1 receptor antagonist, soluble tumor necrosis factor receptor-I or neutrophil gelatinase-associated lipocalin were found between the groups in this study. Mild EIP was accompanied by early leakage of proenzymes and later activation of trypsinogen/proteases. A significant cytokine response and neutrophil activation were recorded the day after ERCP, but further studies are needed to determine the temporal relation between these different pathophysiologic events.


Podocytopathy in Obesity: Challenges of Living Large.
Renal injury resulting from obesity is a growing concern caused by the global obesity epidemic. We discuss the glomerular structure, obesity-related glomerular changes, and diagnostic pathologic criteria for obesity-related glomerulopathy. The three main hypothesized mechanisms of podocyte injury are mechanical stress on the podocytes, metabolic derangement, and genetic/molecular factors. Weight loss, renin-angiotensin-aldosterone system inhibitors, and improved insulin resistance may slow the progression. A more comprehensive understanding of obesity-related glomerulopathy will help in developing more effective therapies.


Potential health benefits of continuous LNG-IUS combined with parenteral ERT for seamless menopausal transition and beyond--a commentary based on clinical experience.
To comment on the acceptability and potential health benefits of the continuous use of the levonorgestrel-releasing intrauterine system (LNG-IUS), combined with estrogen substitution, for seamless transition through the menopause, in women with climacteric symptoms. Evaluation of the recent hormone replacement therapy literature and the acceptability of the combined parenteral estrogen and intrauterine LNG-IUS regimen in a group of approximately 100 women, above 48 years of age, using LNG-IUS for contraception, who developed climacteric symptoms requiring estrogen substitution. acceptability and continued use of the method for the treatment of climacteric symptoms and for prevention. The combination of intrauterine progestogen delivery to suppress the endometrium, in combination with systemic estrogen, is highly acceptable resulting in a high continuation of use due to the absence of side effects and erratic bleeding in the large majority of women. The study suggests that parenteral estrogen replacement therapy combined with intrauterine progestogen delivery for endometrial suppression in the perimenopause is highly practical and beneficial, providing enhanced quality of life. There are strong arguments to categorize the regimen as probably the most effective, safest and best accepted route resulting in high patient compliance as well as potentially providing maximal benefits for peri- and postmenopausal women.