[TRPV4 channel mediates the increase of pulmonary microvascular endothelial permeability in rats with chronic hypoxic pulmonary hypertension].
The purpose of the present study was to investigate the effect of transient receptor potential vanilloid 4 (TRPV4) channel on the permeability of pulmonary microvascular endothelial cells (PMVECs) in rats with chronic hypoxia-induced pulmonary hypertension (CHPH), so as to clarify the mechanism of vascular endothelial dysfunction during the occurrence of pulmonary hypertension (PH). CHPH rat model was established by exposure to chronic hypoxia (CH) for 21 days. Primary PMVECs were cultured by adherent tissue blocks at the edge of the lung. The permeability coefficient of primary cultured PMVECs was detected by fluorescein isothiocyanate (FITC)-dextran. The structure of tight junction (TJ) was observed by transmission electron microscope. The expression of TRPV4 and TJ-related proteins, such as, Occludin, Claudin-5, ZO-1 were examined by real-time fluorescence quantitative PCR and Western blotting. The intracellular calcium concentration ([Ca2+]i) in PMVECs and its effect on PMVECs permeability were observed after the intervention of TRPV4 specific agonist GSK1016790A (GSK, 10 nmol/L) and specific inhibitor HC-067047 (HC, 1 μmol/L, 0.5 μmol/L). The results showed that the CHPH model was successfully established in rats treated with CH for 21 days. In CHPH rats, the structure of TJ was destroyed, the function of PMVECs barrier was decreased, the intercellular permeability was increased, the expression of TJ-related proteins were significantly decreased and the expression of TRPV4 was significantly increased (P < 0.01). The amplitude of [Ca2+]i in PMVECs of CHPH rats was significantly increased after activation of TRPV4. The inhibition ratio of HC on [Ca2+]i in PMVECs of CHPH rats was significantly higher than that in normal PMVECs. TRPV4 specific inhibitor HC reversed the increase of PMVECs permeability and increased the expression of three TJ-related proteins in CHPH rats (P < 0.01, P < 0.05). These results suggest that TRPV4 channel can induce endothelial dysfunction by increasing the [Ca2+]i, resulting in the destruction of TJ structure and the decrease of TJ-related proteins expression on PMVECs in CHPH rats.


Non-typeable Haemophilus influenzae isolates from patients with chronic obstructive pulmonary disease contain new phase-variable modA methyltransferase alleles controlling phasevarions.
Phasevarions (phase-variable regulons) are emerging as an important area of bacterial gene regulation. Many bacterial pathogens contain phasevarions, with gene expression controlled by the phase-variable expression of DNA methyltransferases via epigenetic mechanisms. Non-typeable Haemophilus influenzae (NTHi) contains the phase-variable methyltransferase modA, of which multiple allelic variants exist (modA1-21). We have previously demonstrated 5 of 21 these modA alleles are overrepresented in NTHi strains isolated from children with middle ear infections. In this study we investigated the modA allele distribution in NTHi strains isolated from patients with chronic obstructive pulmonary disease, COPD. We demonstrate that the distribution of modA alleles in a large panel of COPD isolates is different to the distribution seen in middle ear infections, suggesting different modA alleles may provide distinct advantages in the differing niches of the middle ear and COPD airways. We also identified two new phase-variable modA alleles - modA15 and modA18 - and demonstrate that these alleles methylate distinct DNA sequences and control unique phasevarions. The modA15 and modA18 alleles have only been observed in COPD isolates, indicating that these two alleles may be markers for isolates likely to cause exacerbations of COPD.


Acute psychosis in an adolescent with undiagnosed homocystinuria.
Homocystinuria due to cystathionine-β-synthase deficiency (CBS deficiency) usually presents with ectopia lentis, myopia, intellectual disability, skeletal anomalies resembling Marfan syndrome, and thromboembolic events. Whereas neurodevelopment impairments have been often described in untreated homocystinuria adult patients, acute psychosis has rarely been reported as a presenting symptom of the disease. Here, we describe a 17-year-old girl affected by CBS deficiency presenting acute onset of visual hallucinations, behavioral perseverance, psychomotor hyperactivity, and affective inappropriateness. Ectopia lentis, diagnosed several years before, didn't have been considered as possible sign of a metabolic disorder. Psychotic symptoms were unresponsive to the conventional antipsychotic drugs and relieved after pyridoxine and folic acid treatment. A diagnosis of homocystinuria due to CBS deficiency should be considered in patients presenting, as target signs, ectopia lentis with or without learning difficulties, and should also be taken into account as a potentially treatable cause of acute psychosis in childhood and adolescence. • Homocystinuria frequently present with ectopia lentis, myopia, cognitive impairment, Marfan-like phenotype, osteoporosis, cerebrovascular, or cardiac thrombosis. • Acute psychosis has rarely been reported as a presenting symptom of the disease. • The complete psychotic symptoms' remission with pharmacological doses of pyridoxine and folic acid, without antipsychotic drugs.


Clinical evaluation of the Oculus Keratograph.
To determine the validity and reliability of the measurement of corneal curvature and non-invasive tear break-up time (NITBUT) measures using the Oculus Keratograph. One hundred eyes of 100 patients had their corneal curvature assessed with the Keratograph and the Nidek ARKT TonorefII. NITBUT was then measured objectively with the Keratograph with Tear Film Scan software and subjectively with the Keeler Tearscope. The Keratograph measurements of corneal curvature and NITBUT were repeated to test reliability. The ocular sensitivity disease index questionnaire was completed to quantify ocular comfort. The Keratograph consistently measured significantly flatter corneal curvatures than the ARKT (MSE difference: +1.83 ± 0.44D), but was repeatable (p > 0.05). Keratograph NITBUT measurements were significantly lower than observation using the Tearscope (by 12.35 ± 7.45 s; p < 0.001) and decreased on subsequent measurement (by -1.64 ± 6.03 s; p < 0.01). The Keratograph measures the first time the tears break up anywhere on the cornea with 63% of subjects having NITBUTs <5 s and a further 22% having readings between 5 and 10 s. The Tearscope results were found to correlate better with the patients symptoms (r = -0.32) compared to the Keratograph (r = -0.19). The Keratograph requires a calibration off-set to be comparable to other keratometry devices. Its current software detects very early tear film changes, recording significantly lower NITBUT values than conventional subjective assessment. Adjustments to instrumentation software have the potential to enhance the value of Keratograph objective measures in clinical practice.


Evidence of neuroinflammation in fibromyalgia syndrome: a [ 18 F]DPA-714 positron emission tomography study.
This observational study aimed to determine whether individuals with fibromyalgia (FM) exhibit higher levels of neuroinflammation than healthy controls (HCs), as measured with positron emission tomography using [ 18 F]DPA-714, a second-generation radioligand for the translocator protein (TSPO). Fifteen women with FM and 10 HCs underwent neuroimaging. Distribution volume (V T ) was calculated for in 28 regions of interest (ROIs) using Logan graphical analysis and compared between groups using multiple linear regressions. Group (FM vs HC) was the main predictor of interest and TSPO binding status (high- vs mixed-affinity) was added as a covariate. The FM group had higher V T in the right postcentral gyrus ( b = 0.477, P = 0.033), right occipital gray matter (GM; b = 0.438, P = 0.039), and the right temporal GM ( b = 0.466, P = 0.042). The FM group also had lower V T than HCs in the left isthmus of the cingulate gyrus ( b = -0.553, P = 0.014). In the subgroup of high-affinity binders, the FM group had higher V T in the bilateral precuneus, postcentral gyrus, parietal GM, occipital GM, and supramarginal gyrus. Group differences in the right parietal GM were associated with decreased quality of life, higher pain severity and interference, and cognitive problems. In support of our hypothesis, we found increased radioligand binding (V T ) in the FM group compared with HCs in several brain regions regardless of participants' TSPO binding status. The ROIs overlapped with prior reports of increased TSPO binding in FM. Overall, increasing evidence supports the hypothesis that FM involves microglia-mediated neuroinflammation in the brain.


Failure to detect dopamine receptor IgG autoantibodies in sera of schizophrenic patients. Short note.
Autoantibodies against dopamine receptors in schizophrenic patients have been postulated. IgG was fractionated from sera of 15 schizophrenic patients (DSM III) in an acute episode. However, 3H-spiperone binding to dopamine receptors was not inhibited by this fraction.


Head and neck manifestations of tuberculosis.
TB is a serious communicable disease that causes 2 million deaths a year worldwide. It is estimated one third of the world's population is infected with TB. TB has proved difficult to eradicate. Laboratory testing also has been challenging, with as many as 19% of TB cases failing to show positive bacteriology tests. Head and neck masses and oral lesions always should be included in the differential diagnosis of TB, especially in high-risk patient populations. A combination of clinical suspicion, extensive laboratory analysis, and radiographic examination is invaluable in treating and preventing the spread of this insidious but deadly disease that has plagued mankind for ages.


Analgesia (mis)usage on a dental emergency service: a patient survey.
Analgesics are one of the most frequently used medicines. Self-medication and misuse have been described in the literature. The purpose of this study was to document analgesic (mis)use in a population seeking emergency dental treatment. Patients consulting a dental emergency service were randomly asked to complete a questionnaire on analgesic use, knowledge and information on the analgesics and on their pain history. A photobook was used as an aid to identify products used. Descriptive statistics were combined with chi-square and Mann-Whitney U testing. Ninety-eight patients were included. Acetaminophen (69.4%) and ibuprofen (65.3%) were the most frequently used products. Nearly half of the subjects (43.9%) combined at least two analgesics. Although 42.9% of subjects were aware of the maximum daily dose, 62.2% of the subjects exceeded this limit, specifically 76.6% of subjects using ibuprofen and 32.4% of subjects using acetaminophen overdosing. Females overdosed significantly more than males. Ingestion on medical advice did not affect the overdose rates significantly. No significant relation was found between the absence of knowledge on the maximum daily dose and actual overdosing. No higher pain reduction was found in patients overdosing analgesics. The average number of days patients experienced pain before consulting the emergency unit was 12. A significant relation was found between the lag time and overdosing. A large portion of the patients overdosed analgesics. Even prior medical advice did not reduce significantly overdose rates. Dentists treating emergency cases clearly need to be aware of the high risk and high rates of overdosing analgesics in their patients.


Staging laparotomy for inoperable or borderline operable cancer of the rectum.
The authors report their experience with the staging laparotomy as a means of identifying and preparing patients for high-dose preoperative radiotherapy. Twelve patients had clinically unresectable cancers of the rectum. The goal of the staging laparotomy is to assess mobility and tumor size by means of bimanual palpation, to stage the abdominal cavity, and to fashion an end colostomy at the level of the descending colon. Eight patients ultimately underwent radical resection. Three died during follow-up due to hematogenic metastases without recurrent pelvic disease. Five patients are alive with no evidence of disease and have been followed for an average of 34 months (range, 20 to 64 months).


Infectious bursal disease viral infections. II. The relationship of age, complement levels, virus-neutralizing antibody, clotting, and lesions.
Experimental infection with infectious bursal disease virus (IBDV) reduced the complement (C) titer in 8-week-old chickens on days 3, 5, and 7 postinfection. Since the C titer was much lower in normal 2-week-old chickens than in normal 8-week-old chickens it could not be determined whether there was a reduction in titer during the infection process. Virus-neutralizing antibody rose rapidly following infection in both 2- and 8-week-old chickens. Hyper-immune serum given during infection in an attempt to produce immune complexes did not increase disease severity in either 2- or 6-week-old susceptible chickens.


Amino acid sequence of alpha-chain of hemoglobin IV from trout (Salmo irideus).
The amino acid sequence of the alpha-chain of trout hemoglobin (Hb) IV is given, thus completing the primary structure of the hemoglobin component of trout's blood characterized by the Root effect. The trout Hb IV alpha-chain consists of 142 amino acid residues; comparison with the corresponding sequences from human and carp hemoglobins shows differences of 50.0 and 35.9%, respectively. A difference of 39.6% is found with the alpha-chain of trout Hb I, the other major hemoglobin component of trout blood, devoid of heterotropic effects.


Gelatin/monetite electrospun scaffolds to regenerate bone tissue: Fabrication, characterization, and in-vitro evaluation.
This work is dedicated to combining nanotechnology with bone tissue engineering to prepare and characterize electrospun gelatin/monetite nanofibrous scaffold with improved physicochemical, mechanical, and biological properties. Nanofibrous scaffolds possessing fiber diameter in the range of 242-290 nm were prepared after incorporating varying content of monetite nanoparticles up to 7 wt % into the gelatin matrix using the electrospinning technique. Cross-linking of gelatin chains in the scaffold was performed using 0.25 wt% glutaraldehyde as indicated by imine (-CN-) bond formation in the FTIR analysis. With an increase in monetite addition up to 7 wt%, a decrease in swelling ratio and bio-degradability of cross-linked gelatin scaffolds was observed. Gelatin scaffold with 7 wt% monetite content registered the highest values of tensile strength and tensile modulus of 18.8 MPa and 170 MPa, as compared to 0% and 5 wt% monetite containing scaffolds respectively. Cell viability and differentiation were studied after culturing MG-63 cells onto the scaffolds from confocal microscopy of live and dead cells images, MTT assay, and alkaline phosphatase assay for a cell culture period of up to 21 days. It was observed that 7 wt % monetite containing gelatin scaffold exhibited better MG-63 cell adhesion, proliferation, higher biomineralization, and ALP activity compared to 0% and 5 wt% monetite containing electrospun scaffolds studied here.


A study of IgM rheumatoid factors in a middle-aged population of Northern Norway.
A random selection of 8807 blood samples was analysed for the presence of IgM rheumatoid factor (RF). The sera were obtained from an epidemiological survey of rheumatoid arthritis in a middle-aged population of Northern-Norway. The prevalence of IgM-RF was 1.36% for the total population, but a prevalence of IgM RF in healthy individuals between 0.48-0.49% was found with no significant difference between males and females. The majority of RF positive sera were low titred. Only 11.4% of the persons possessing serum IgM RF suffered from classical or definite rheumatoid arthritis. Eighty-one per cent of the IgM RF positive individuals converted to seronegativity in the course of 3.5 years.


Purification and partial characterization of a lectin from the seeds of Trichosanthes kirilowii Maximowicz.
A lectin was purified from the seeds of Trichosanthes kirilowii, belonging to the family Cucurbitaceae, growing in China. The lectin is a glycoprotein of 57 kDa, consists of two subunits with apparent molecular masses of 37 and 25 kDa, is specific for galactose, and is not mitogenic for human lymphocytes.


Relative bioavailability of 3 rumen-undegradable methionine sources in dairy cows using the area under the curve technique.
The objective of this study was to evaluate the relative bioavailability of two 2-hydroxy-4-(methylthio)butanoic isopropyl esters (HMBi) obtained through different production processes and an encapsulated rumen-protected Met using the area under the curve (AUC) method. The new HMBi product (Kessent MF Liquid, Kemin Animal Nutrition and Health) was compared with an existing HMBi product (Metasmart, Adisseo SAS) and a pH-sensitive coated Met (Smartamine, Adisseo SAS). Nine multiparous lactating cows (30 kg of milk/d and 227 d in milk) fed a 45:55 forage:concentrate diet were randomly assigned within square to a triplicate 3 × 3 Latin square design. Each period consisted of a 3-d sampling period and a 3-d washout period. Treatments were dosed on d 1 of each period, and blood samples were collected from the coccygeal vein at 0, 1, 2, 3, 4, 6, 9, 12, 24, 30, and 48 h thereafter. The daily dose was 50 g of Met equivalent of each treatment. The HMBi treatments were administered directly into the cow's mouth, whereas Smartamine was fed mixed with 0.5 kg of concentrate and fully consumed within 15 min. Nonlinear models were fitted to raw data, and the basal concentration at time 0 h, time at peak (Tmax), concentration at peak, and AUC of plasma Met were determined. The Met basal concentration at t = 0 h (26.7 ± 7.67 µM) and concentration at peak (210 ± 22.2 µM) were similar among treatments, but the Tmax (11.3 vs. 1.4 h) was delayed and the AUC was 1.8-fold larger (3,457 vs. 1,868 arbitrary units) in Smartamine compared with HMBi. Results of this study indicate that the 2 HMBi products have similar plasma kinetics and bioavailability. Smartamine had different kinetics compared with HMBi products, with delayed Tmax and larger AUC and relative bioavailability.


The 1982 Medicaid Funding Cessation for Circumcision in California and Circumcision Rates.
We investigated California's 1982 decision to stop funding Medicaid neonatal circumcision. We examined male neonatal circumcision rates for those born 1977-1981 and 1983-1987 by region, race, and insurance status. Overall, West-Medicaid circumcision rates decreased from 56.5% in 1979-81 to 26.7% in 1983-85. California's 1982 decision to defund Medicaid circumcision coverage was associated with a 25.0-30.8% point decrease in West-Medicaid circumcision rates compared other groups, p < 0.01. This provides the earliest data to support that funding coverage for neonatal circumcision affects circumcision rates and magnifies healthcare disparities. Other states have since defunded Medicaid male neonatal circumcision. Circumcision have been associated with lower rates of sexually transmitted infections including HIV, and urinary tract infections. Lawmakers should consider re-funding Medicaid male neonatal circumcision.


Fast segmentation of anterior segment optical coherence tomography images using graph cut.
Optical coherence tomography (OCT) is a non-invasive imaging system that can be used to obtain images of the anterior segment. Automatic segmentation of these images will enable them to be used to construct patient specific biomechanical models of the human eye. These models could be used to help with treatment planning and diagnosis of patients. A novel graph cut technique using regional and shape terms was developed. It was evaluated by segmenting 39 OCT images of the anterior segment. The results of this were compared with manual segmentation and a previously reported level set segmentation technique. Three different comparison techniques were used: Dice's similarity coefficient (DSC), mean unsigned surface positioning error (MSPE), and 95% Hausdorff distance (HD). A paired t-test was used to compare the results of different segmentation techniques. When comparison with manual segmentation was performed, a mean DSC value of 0.943 ± 0.020 was achieved, outperforming other previously published techniques. A substantial reduction in processing time was also achieved using this method. We have developed a new segmentation technique that is both fast and accurate. This has the potential to be used to aid diagnostics and treatment planning.


Erratum: Comparing endoscopic thyroidectomy using the breast approach and conventional open thyroidectomy: A retrospective analysis.
[This corrects the article DOI: 10.4103/jcrt.jcrt_707_21].


Metabolic danger signals, uric acid and ATP, mediate inflammatory cross-talk between hepatocytes and immune cells in alcoholic liver disease.
Inflammation defines the progression of ALD from reversible to advanced stages. Translocation of bacterial LPS to the liver from the gut is necessary for alcohol-induced liver inflammation. However, it is not known whether endogenous, metabolic danger signals are required for inflammation in ALD. Uric acid and ATP, 2 major proinflammatory danger signals, were evaluated in the serum of human volunteers exposed to a single dose of ethanol or in supernatants of primary human hepatocytes exposed to ethanol. In vitro studies were used to evaluate the role of uric acid and ATP in inflammatory cross-talk between hepatocytes and immune cells. The significance of signaling downstream of uric acid and ATP in the liver was evaluated in NLRP3-deficient mice fed a Lieber-DeCarli ethanol diet. Exposure of healthy human volunteers to a single dose of ethanol resulted in increased serum levels of uric acid and ATP. In vitro, we identified hepatocytes as a significant source of these endogenous inflammatory signals. Uric acid and ATP mediated a paracrine inflammatory cross-talk between damaged hepatocytes and immune cells and significantly increased the expression of LPS-inducible cytokines, IL-1β and TNF-α, by immune cells. Deficiency of NLRP3, a ligand-sensing component of the inflammasome recognizing uric acid and ATP, prevented the development of alcohol-induced liver inflammation in mice and significantly ameliorated liver damage and steatosis. Endogenous metabolic danger signals, uric acid, and ATP are involved in inflammatory cross-talk between hepatocytes and immune cells and play a crucial role in alcohol-induced liver inflammation.


Peculiar angiographic predictors of impending left ventricular rupture after primary coronary angioplasty.
Peculiar angiographic characteristics predictive of impending left ventricular rupture have not yet been described in primary percutaneous coronary angioplasty era. Our study is aimed to evaluate the existence of peculiar angiographic determinants of impending left ventricular rupture after primary percutaneous coronary angioplasty. We retrospectively analyzed medical and angiographic records of patients referred to our center for acute myocardial infarction (AMI) and submitted to primary angioplasty between 1st May 2005 and 30th September 2007. All the coronary angiograms and coronary angioplasties were reviewed by two independent observers blinded to the final outcome of the patients searching for any abnormal angiographic signs of impending left ventricular rupture. The records of 348 patients (mean age, 76+/-25.8 years, 198 females) with AMI submitted to primary percutaneous coronary interventions (PCI) have been reviewed and analyzed. Left ventricular rupture occurred in 11 patients (3.16%): 10 of 11 patients died, whereas one survived to an inferoposterior left ventricular rupture conservatively managed with repeated pericardiocentesis and blood transfusions. The observers found in all 10/11 cases (2.87%) of left ventricular ruptures some angiographic signs of impending rupture such as round-shaped contrast persistence in the region of the rupture confirmed by autopsy or in the only one survivor by computed tomography. No similar images were detected on angiography of the remaining 337 patients (P<.001). Logistic regression analysis demonstrated that only long time that elapsed from symptoms onset, cardiogenic shock, and abnormal angiographic contrast dye persistence images were independent predictors of cardiac rupture. Angiographic signs of impending ventricular rupture can be observed during primary coronary angioplasty and could help physicians to adopt a more accurate echocardiographic protocol in patients with such signs in order to refer patients without delay to surgical repair.


Use of deferiprone for iron chelation in patients with transfusion-dependent thalassaemia.
To conduct a retrospective case analysis of the clinical efficacy and adverse effects of deferiprone in our population. All patients with transfusion-dependent thalassaemia at KK Hospital who have been on deferiprone were included in the study. Outcomes measured include the change in ferritin levels and cardiac T2* values during deferiprone therapy, and incidence of side effects. Thirty-three (47.1%) of the total cohort of 70 patients have been on deferiprone, out of which 26 were on combination therapy with desferrioxamine. Majority of the patients (76%) had stable cardiac iron load during deferiprone therapy, and four patients with moderate to severe cardiac iron load showed improvement. Ten patients (30.3%) had improvement in their ferritin levels. Three patients (9.1%) developed mild neutropenia at 3, 18 and 26 months, respectively, and two patients (6.1%) had agranulocytosis at 4 and 10 months, respectively. Their neutrophil counts improved spontaneously after cessation of deferiprone. Thrombocytopenia developed in 27.3% of the patients and was transient in majority (77.8%) of the patients. Five patients (15.2%) developed arthritis that improved after cessation of deferiprone therapy, and one patient had transient arthralgia that resolved spontaneously. Three patients (9.1%) had nausea and abdominal pain. Deferiprone effectively reduced or stabilised cardiac iron load in our patients. Thrombocytopenia, arthropathy, neutropenia and agranulocytosis are the most important side effects. It is recommended that patients on deferiprone have their full blood counts monitored weekly for the first year of therapy and subsequently fortnightly as long as they are on deferiprone.


A Platinum(II)-Based Molecular Cage with Aggregation-Induced Emission for Enzymatic Photocyclization of Alkynylaniline.
Enzymes facilitate chemical conversions through the collective activity of aggregated components, but the marriage of aggregation-induced emission (AIE) with molecular containers to emulate enzymatic conversion remains challenging. Herein, we report a new approach to construct a PtII -based octahedral cage with AIE characteristics for the photocyclization of alkynylaniline by restricting the rotation of the pendant phenyl rings peripheral to the PtII corner. With the presence of water, the C-H⋅⋅⋅π interactions involving the triphenylphosphine fragments resulted in aggregation of the molecular cages into spherical particles and significantly enhanced the PtII -based luminescence. The kinetically inert Pt-NP chelator, with highly differentiated redox potentials in the ground and excited states, and the efficient coordination activation of the platinum corner facilitated excellent catalysis of the photocyclization of alkynylaniline. The enzymatic kinetics and the advantages of binding and activating substrates in an aqueous medium provide a new avenue to develop mimics for efficient photosynthesis.


Condensation reactions of guanidines with bis-electrophiles: Formation of highly nitrogenous heterocycles.
2-Amino-1,4-dihydropyrimidines were reacted with bis-electrophiles to produce novel fused bi-pyrimidine, pyrimido-aminotriazine, and pyrimido-sulfonamide scaffolds. In addition, a quinazoline library was constructed using a guanidine Atwal-Biginelli reaction with 1-(quinazolin-2-yl)guanidines. The product heterocycles have novel constitutions with high nitrogen atom counts and represent valuable additions to screening libraries for the discovery of new modulators of biological targets.


Inference of genotype-phenotype relationships in the antigenic evolution of human influenza A (H3N2) viruses.
Distinguishing mutations that determine an organism's phenotype from (near-) neutral 'hitchhikers' is a fundamental challenge in genome research, and is relevant for numerous medical and biotechnological applications. For human influenza viruses, recognizing changes in the antigenic phenotype and a strains' capability to evade pre-existing host immunity is important for the production of efficient vaccines. We have developed a method for inferring 'antigenic trees' for the major viral surface protein hemagglutinin. In the antigenic tree, antigenic weights are assigned to all tree branches, which allows us to resolve the antigenic impact of the associated amino acid changes. Our technique predicted antigenic distances with comparable accuracy to antigenic cartography. Additionally, it identified both known and novel sites, and amino acid changes with antigenic impact in the evolution of influenza A (H3N2) viruses from 1968 to 2003. The technique can also be applied for inference of 'phenotype trees' and genotype-phenotype relationships from other types of pairwise phenotype distances.


T testing the immune system.
Amid the flurry of grant writing and experimentation, statistical analysis sometimes gets less attention than it requires. Here, we describe fully the considerations that should go into the employment of the statistical two-sample t test.


Effects of administration styles of arsenic trioxide on intracellular arsenic concentration, cell differentiation and apoptosis.
We studied the effects of varying and steady-state concentrations of arsenic trioxide (As2O3) on apoptosis and differentiation of several cell lines in vitro. We also studied the same effects of fluctuating vs constant concentrations of As2O3 in vivo in patients treated with daily 3-hour fast infusions or daily slow, continuous infusions. Intracellular concentrations of arsenic and apoptosis rate were higher whereas differentiation was reduced in cells exposed to constant concentrations of As2O3.


Acute chiasmal abscess resulting from perineural extension of invasive sino-orbital aspergillosis in an immunocompetent patient.
Intracranially invasive sino-orbital aspergillosis is a rare entity seen predominantly in immunocompromised individuals. We report a unique case of an acute chiasmal abscess resulting from perineural extension of an indolent invasive sino-orbital aspergillosis in an immunocompetent patient. A 61-year-old healthy Ukrainian man presented with severe left retro-orbital pain and a gradual ipsilateral monocular vision loss with rapid progression to a contralateral temporal visual field cut. He was found to have an intracranially invasive sino-orbital Aspergillus fumigatus infection with perineural extension along the optic nerve complicated by an acute chiasmal abscess. The patient was managed with surgical debridement and adjuvant antifungal chemotherapy. The patient's life and vision were preserved. The case demonstrates that it is possible to successfully control advanced intracranial aspergillosis with tissue-sparing surgery and adjuvant antifungal chemotherapy in immunocompetent individuals.


Excess liver transplant waitlist mortality for patients with primary biliary cholangitis under MELD-Na allocation.
Historically, patients with primary biliary cholangitis (PBC) experience waitlist mortality and low rates of liver transplant (LT). Herein, the impact of MELD-Na based allocation on PBC waitlist mortality was examined. Adult patients with PBC were compared to those with alcohol-related liver disease (ALD) or non-alcoholic steatohepatitis (NASH) listed for LT from 2013 to 2019 in OPTN. Competing risk regression evaluated waitlist mortality in the MELD and MELD-Na eras using propensity score weights. Overall, 1508 patients with PBC, 13581 with ALD, and 10455 with NASH were examined. In the MELD-Na era, 24-month cumulative incidence of waitlist mortality for PBC was 23.0% (95%CI 19.7-26.5%), ALD 13.9% (95%CI 13.1-14.8%), and NASH 20.0% (95%CI 18.9-21.2%). Using propensity score weights, adjusted risk of waitlist mortality was higher for PBC versus ALD (HR = 1.45, 95%CI 1.22-1.71) and NASH (HR = 1.32, 95%CI 1.14-1.55). Furthermore, among PBC, waitlist mortality risk per five-point elevation in MELD-Na (HR = 1.22, 95%CI 1.11-1.35) and Karnofsky score ≤30% (HR = 2.02, 95%CI 1.39-2.92) was significantly higher than among ALD (HR = 1.08, 95%CI 1.04-1.13; HR = 1.28, 95%CI 1.10-1.49) and NASH (HR = 1.05, 95%CI 1.00-1.09; HR = 1.16, 95%CI .99-1.37; all P-interactions < .05). The MELD-Na score continues to underestimate risk of waitlist death for patients with PBC relative to ALD and NASH and highlights need for additional score modifications or exceptions.


Copper oxide nanoparticle induces inflammatory response and mucus production via MAPK signaling in human bronchial epithelial cells.
Copper nanoparticles (CuONPs) can pose risks to industrial workers. With increase of its applications especially in electronic fields, it is necessary to assess the toxicity of CuONPs, including pulmonary toxicity. In this study, we investigated the effect of CuONPs on human epithelial cell line H292. CuONPs treatment caused a significant increase in IL-6 and IL-8 mRNA expression and protein levels in H292 cells in a concentration-dependent manner. The mRNA expression and protein levels of MUC5AC were consistent with those of proinflammatory mediators. Additionally, CuONPs treatment increased phosphorylation of mitogen-activated protein kinases (MAPKs), Erk, JNK, and p-38 compared to that of control in a concentration-dependent manner. However, co-treatment with CuONPs and each MAPK inhibitor significantly decreased the phosphorylation of each MAPK, resulting in decreased mRNA expression and protein levels of proinflammatory mediators and MUC5AC compared to that in H292 cells only treated with CuONPs. In summary, CuONPs-induced inflammatory mediators and MUC5AC associated with MAPKs phosphorylation. Our results will provide useful information on CuONPs-induced pulmonary toxicity.


Biocompatible Nanotube-Strontium/polydopamine-arginine-glycine-aspartic acid coating on Ti6Al4V enhances osteogenic properties for biomedical applications.
Titanium (Ti) alloys, particularly Ti6 Al4 V, are the most commonly used biomedical implant material. Ti alloys are biologically inert, so there have been continuous efforts to improve their osteogenic properties and clinical performance. Since TiO2 nanotubes (NT) appear to be excellent drug platforms, and strontium reportedly enhances osteogenesis, we constructed a TiO2 nanotube coating on the surface of Ti6 Al4 V and immersed it in Sr (OH)2 solution in order to incorporate Sr into TiO2 nanotubes (NT-Sr). The results of field emission scanning electron microscope and X-ray diffraction analysis verified the fabrication of NT-Sr. We next added polydopamine (PDA) and cyclo- (arginine-glycine-aspartic acid-phenylalanine-cysteine) [c(RGDfC)] peptides to further promote biocompatibility of the implant. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy confirmed the existence of PDA and c(RGDfC). Mesenchymal stem cells (MSCs) were planted on Ti, NT, NT-Sr, NT-Sr/PDA, and NT-Sr/PDA-RGD surfaces. The adhesion and differentiation of MSCs on different surfaces were evaluated. The mRNA expression of alkaline phosphatase, runt-related transcription factor 2 (Runx2) and type I collagen (Col I) of different groups were also tested. Finally, we observed that the NT-Sr/PDA-RGD group showed significantly better performance than other groups in terms of the differentiation and osteogenesis-related gene expression of MSCs. Thus, the NT-Sr/PDA-RGD complex may be an important modification strategy for Ti, as it shows excellent osteogenic potential.


Evaluation of different API systems for identification of porcine Pasteurella multocida isolates.
An exhaustive biochemical characterisation of 60 porcine Pasteurella multocida clinical isolates recovered from lesions indicative of pneumonia, previously confirmed by PCR and all belonging to the capsular serogroup A, was performed by means of four commercial systems. The API 20NE correctly identified almost all isolates (95%), but only 60% could be ascribed to this species by the API 20E method. The high diversity exhibited by the API 50CHB/E system, with six different patterns, does not advise its use as additional system for a definitive identification at the species level, but this method could be a potential tool for characterising P. multocida isolates below this level. The more uniform reactions yielded by the API ZYM test make this system helpful in the confirmatory identification of this organism. The high variability (20 profiles) obtained when the four systems are taken together also suggests their usefulness for epidemiological purposes in order to sub-type P. multocida isolates.


Hepatocellular necrosis, apoptosis, and proliferation after transcatheter arterial embolization or chemoembolization in a standardized rabbit model.
To evaluate the effect of transcatheter arterial chemoembolization versus transcatheter arterial embolization on hepatocellular damage, apoptosis, proliferation, and proinflammatory response in a rabbit VX2 tumor model. Rabbits implanted with VX2 tumors in left liver lobes were randomly divided into three groups: a control group (n = 9) that underwent infusion of distilled water into the left hepatic artery, an embolization group (n = 15) that underwent left hepatic artery embolization with polyvinyl alcohol (PVA) particles, and a chemoembolization group (n = 15) that underwent left hepatic artery infusion of a mixture of 10-hydroxycamptothecin and iodized oil followed by PVA embolization. Serum and liver samples were collected at 6 hours, 3 days, and 7 days postoperatively. Liver damage was measured by liver function tests and histologic analysis. Ki-67 immunohistochemistry and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling were performed to quantify proliferating and apoptotic cells. Serum tumor necrosis factor (TNF)-α levels were measured to assess proinflammatory response. Compared with embolization, chemoembolization caused liver injury with a greater increase in serum alanine aminotransferase and aspartate aminotransferase levels on days 3 and 7; histologic analysis showed increased hepatic necrosis in adjacent liver tissue beginning at day 3 and increased serum levels of TNF-α at 6 hours. By contrast, chemoembolization resulted in a slower increase in hepatocyte proliferation. Additionally, increased apoptotic hepatocytes were observed after embolization and chemoembolization. In contrast to embolization, nonsuperselective transcatheter arterial chemoembolization increased hepatocellular damage and stimulated systemic proinflammatory cytokine release, but inhibited hepatocyte proliferation.


Final analysis of a phase II study of nivolumab in combination with ipilimumab for unresectable chemotherapy-naive advanced melanoma.
Nivolumab plus ipilimumab combination is currently one of the preferred regimens for advanced melanoma in recently updated clinical practice guidelines. However, the evidence on the efficacy of the combination for acral or mucosal subtypes remains less robust. This is the final analysis of a multicenter, open-label, uncontrolled phase II study that investigated the long-term efficacy and safety in treatment-naive Japanese patients with advanced melanoma, including acral or mucosal subtypes, and subsequent therapy after discontinuation of the investigational agents. Patients received four doses of nivolumab (1 mg/kg i.v.) in combination with ipilimumab (3 mg/kg i.v.) at 3-week intervals, followed by doses of nivolumab (3 mg/kg i.v.) at 2-week intervals. The median follow-up period was 20.8 months (range, 5.2-35.0). The centrally and locally assessed objective response rates were both 43.3% (13/30; 95% confidence interval [CI], 25.5-62.6). Median progression-free survival was not reached (95% CI, 3.02-not reached), and median overall survival was also not reached (95% CI, 19.52-not reached). The 30-month progression-free survival and overall survival rates were 50.3% and 54.2%, respectively. No new safety concerns were detected. After discontinuation of the investigational agents, 83.3% of patients received some form of subsequent therapy including 43.3% of patients who received nivolumab monotherapy and 26.7% of patients who received radiotherapy. Of the four patients who discontinued the investigational agents because of immune-related adverse events, two received subsequent therapy (nivolumab and ipilimumab, respectively) and the other two showed long-term treatment-free survival (659 and 590 days, respectively). Long-term survival with nivolumab plus ipilimumab was observed in Japanese patients with melanoma including acral and mucosal subtypes, which is consistent with the CheckMate 067 study. Many patients continued to receive some form of treatment safely after stopping treatment with nivolumab plus ipilimumab.


Antifungal therapy: Novel drug delivery strategies driven by new targets.
In patients with compromised immunity, invasive fungal infections represent a significant cause of mortality. Given the limited availability and drawbacks of existing first-line antifungal drugs, there is a growing interest in exploring novel targets that could facilitate the development of new antifungal agents or enhance the effectiveness of conventional ones. While previous studies have extensively summarized new antifungal targets inherent in fungi for drug development purposes, the exploration of potential targets for novel antifungal drug delivery strategies has received less attention. In this review, we provide an overview of recent advancements in new antifungal drug delivery strategies that leverage novel targets, including those located in the physio-pathological barrier at the site of infection, the infection microenvironment, fungal-host interactions, and the fungal pathogen itself. The objective is to enhance therapeutic efficacy and mitigate toxic effects in fungal infections, particularly in challenging cases such as refractory, recurrent, and drug-resistant invasive fungal infections. We also discuss the current challenges and future prospects associated with target-driven antifungal drug delivery strategies, offering important insights into the clinical implementation of these innovative approaches.


Gender differences in the association of epicardial adipose tissue and coronary artery calcification: EPICHEART study: EAT and coronary calcification by gender.
The association of epicardial adipose tissue (EAT) and coronary artery calcification (CAC) seems to differ by gender. However, few studies have controlled for body size, and the ideal method for body size indexing has not been explored. To analyse the effect of gender related-body size and-body fat differences on the association of EAT with CAC. This was a prospective cohort of 371 severe aortic stenosis patients (77±8.5year-old, 51% females) referred to cardiac surgery. Agatston calcium score, EAT volume and visceral abdominal fat (VAF) were obtained by computed tomography. Body composition was determined using bioelectrical impedance analysis. Body weight and height were measured to derive body mass index (BMI), body surface area (BSA), and body surface index (BSI). EAT volume was normalized for BSA, weight and height. Median CAC score was higher in men (887; IQR: 2010) than in women (279: IQR: 145) (p<0.01). Similarly, men had higher volume of EAT than women (137±65.6 vs. 106±65.6mL, p<0.01), even when BSA- or height-indexed, but not if weight-indexed. EAT volume was associated with CAC adjusting for adiposity (BMI or BSI and VAF, or fat mass), but not with further adjustment for gender. In a stratified analysis, absolute- and indexed-volumes of EAT were independently associated with CAC in men while no association was found in women (gender-interaction p<0.05). In these high-risk patients, we demonstrated that EAT was associated with CAC score irrespective of body size, body fat and cardiovascular risk factors in men but not in women.


Expression of a novel cytokine, IL-4delta2, in HIV and HIV-tuberculosis co-infection.
Correcting the Th2 shift in HIV/AIDS represents a potential intervention strategy. However data on interleukin (IL)-4 expression in HIV or AIDS are un-interpretable because of failure to distinguish between IL-4 and its splice variant and natural antagonist, IL-4delta2. To determine Th1 [interferon (IFN)-gamma], IL-4delta2 and Th2 (IL-4) expression in whole blood and lung lavage from healthy volunteers and in HIV or HIV-tuberculosis (TB) co-infection. Cross-sectional with prospective cohort. Expression of IL-4delta2, IL-4 and IFN-gamma were determined by quantitative real-time PCR, using unstimulated cells from whole blood and lung lavage, in 20 HIV-TB (pulmonary) co-infected patients, 20 matched HIV-positive controls and 20 HIV-negative healthy volunteers. Results were correlated with plasma viral load, CD4 cell counts, radiological scores and response to anti-TB treatment. Compared to HIV negative donors, stable HIV-positive donors did not have increased levels of mRNA encoding IL-4, IL-4delta2 or IFN-gamma in blood or lavage. By contrast, the HIV-TB co-infected donors had increased IL-4 and IFN-gamma in both compartments. However the antagonist, IL-4delta2 was increased only in lavage. Consequently the dominant form was IL-4delta2 in lavage, but IL-4 itself in blood. The lung IL-4/IFN-gamma ratio correlated with radiological disease extent. With anti-TB treatment, IL-4 levels did not change whilst IL-4delta2 levels increased significantly. IL-4 and its natural antagonist, IL-4delta2 and are not upregulated in the absence of opportunistic infection. However in HIV-TB co-infection both cytokines increase in lung, but only IL-4 in the periphery. Further studies are required to determine if IL-4 facilitates systemic HIV progression.


Meat differentiation between pasture-fed and concentrate-fed sheep/goats by liquid chromatography quadrupole time-of-flight mass spectrometry combined with metabolomic and lipidomic profiling.
Animal feeding method is a crucial factor in influencing meat quality. Consumers would preferentially select meat obtained from pasture-fed animals. In this study, an untargeted metabolomic and lipidomic method based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) combined with chemometric analysis was utilized to investigate the differences between meat from free-range and intensively-fed sheep/goats. Distinct separation between these two kinds of sheep/goats meat obtained were identified by principal component analysis. Analysis of variance, fold change and orthogonal projection to latent structures discriminant analysis were then conducted to determine specific potential markers. A total of 46 potential markers were selected according to online chemical databases. The support vector machine (SVM) method was used to process the responses of the selected potential markers, and the results of metabolomics and lipidomics from an additional 59 samples revealed the discrimination rate of 89.3% and 98.3%. These findings provided a basis for differentiation of meat from sheep/goats fed in the two methods.


Antibodies against murine typhyus in sera from Indonesians.
A sero-epidemiological study of human CF and indirect immunofluorescence antibodies against murine typhus was conducted in Indonesia. Antibody prevalences ranged from 10% to 20% in sera of Indonesians from Java and Sumatra to less than 2% in sera from islands east of Java.


Anomalous hygroscopic growth of fine particles of MgSO4 aerosols investigated by FTIR/ATR spectroscopy.
This paper demonstrates an approach for investigating the hygroscopic growth of MgSO4 aerosols deposited on ZnSe substrate by using attenuated total reflection Fourier transform infrared (FTIR/ATR) spectroscopy. The experimental setup based upon a refitted standard FTIR/ATR accessory was adopted for the hygroscopic study of aerosols. It has been found that the "predeliquescence" of fine MgSO4 aerosol particles with the mean spreading diameter around 500 nm occurred before roughly 15% RH. In contrast, the abrupt water absorption of coarse MgSO4 particles with the mean diameter larger than about 10 microm was reported to occur at roughly 42% RH, which was reproduced in this study. Up to now, both theoretical and experimental investigations were rare and immature for fine particles, but the method we used in this study worked very well for the fine particles of MgSO4 aerosols. It has been found that the possible reason for the "predeliquescence" fine particles of MgSO4 aerosols is that the initial state or phase of fine particles is different from coarse ones after desiccation. This research demonstrates that the approach based on the ATR technique is very convenient, accurate and requires only a little amount of lab supplies.


Hemolytic anemia after mitral valve repair: a case report.
Hemolytic anemia after mitral valve repair is still an underestimated complication because it is a rare condition and there are few described case reports in the literature. The mechanism responsible for hemolysis most commonly involves a regurgitant jet and it appears to be independent of its severity as assessed by echocardiography. Patients may experience severe symptoms with only moderate regurgitant jets. We present a case of a 74-year-old Caucasian female who developed severe hemolytic anemia and decompensated heart failure due to a moderate mitral valve regurgitation jet, but it was not considered severe enough to explain such hemolysis. After exclusion of other causes of hemolytic anemia and the lack of clinical and laboratory improvement, the patient underwent valve replacement with a mechanical valve. Anemia and heart failure symptoms gradually resolved after surgery. This case report indicates the importance of including the diagnosis of hemolytic anemia after mitral valve repair in patients with severe hemolysis and a surgical history of heart surgery, even if echocardiography underestimates or is unclear for showing significant alterations. The interest on mitral valve repair as suggested by the increase in the number of these procedures performed worldwide, raises the possibility that hemolytic anemia could be probably seen more frequently in the future and be a cause of major concern, especially for cardiac surgeons.


The potential role of eyestalk in the immunity of Litopenaeus vannamei to Vibrio parahaemolyticus infection II. From the perspective of long non-coding RNA.
Long non-coding RNAs (lncRNAs) have been linked to immunological modulation. Unfortunately, little is known about the processes of immune control in shrimp. In crustaceans such as Litopenaeus vannamei, a prominent aquaculture species, the X-organ-sinus gland complex (XO-SG) in the eyestalk is an essential neuroendocrine regulatory organ. Eyestalk ablation is commonly employed in aquaculture to accelerate ovarian maturation in shrimp. It does, however, have a negative impact on the shrimps' immunocompetence and causes death. As a result, we used RNA-seq to profile the transcriptomes of L. vannamei hemocytes infected with Vibrio parahaemolyticus after the eyestalk ablation. Following strict transcript screening procedures, 2307 lncRNAs were identified from L. vannamei hemocytes in this study. Pearson correlation analysis was finally used to uncover 535 DElncRNAs and 1566 DEmRNA targets. According to the Venn diagram analysis, 326 non-eyestalk regulatory lncRNAs (NElncRNAs) with a target of 1014 non-eyestalk regulatory genes (NEmRNAs), 47 eyestalk negative regulatory lncRNAs (ENRlncRNAs) with a target of 95 eyestalk negative regulatory genes (ENRmRNAs), and 162 eyestalk positive regulatory lncRNAs (EPRlncRNAs) with a target of 457 eyestalk positive regulatory genes (EPRmRNAs) were screened. The bioinformatics analysis revealed that lncRNAs were associated with Axon regeneration, Rap1 signaling pathway, Thyroid hormone signaling pathway, TGF-beta signaling pathway, and PI3K-Akt signaling pathway, implying that lncRNAs may play a role in the regulation of the neuroendocrine-immune (NEI) system. Furthermore, several lncRNAs targeting HSP70, YWHAZ, FER2, HIF1α, and Notch were discovered and verified by qRT-PCR. These findings showed that regulation of lncRNAs in hemocytes which were controlled by the eyestalk might be one of the impact variables in controlling the differential expression of mRNAs associated with immune response in L. vannamei infected with V. parahaemolyticus.


A solvent-free approach to extract the lipid fraction from sewer grease for biodiesel production.
Fats, oils and greases (FOG) are the number one cause of sewer pipe blockage and have been mostly disposed of as a waste until recently. This study investigated a low cost and environmentally friendly approach to extract the lipid fraction (fatty acids and glycerides for biodiesel production) from sewer grease (SG), i.e., FOGs obtained from wastewater treatment plants (WWTPs). The lipid fraction of the sewer grease was primarily in the form of free fatty acid (FFA), at 20.7wt%. An innovative solvent-free extraction approach was developed using waste cooking oil (WCO) to overcome the challenges of emulsion, impurities and high moisture content of the sewer grease. A 95% extraction yield of sewer grease was achieved under the optimum operating condition of 3.2:1 WCO-SG ratio (wt/wt), 70°C and 240min. In addition, the reusability of the WCO was also investigated. WCO can be used two to three times for sewer grease extraction with more than 90% extraction efficiency.


Synergistic Effect of Simultaneous Doping of Ceria Nanorods with Cu and Cr on CO Oxidation and NO Reduction.
Ceria particles play a key role in catalytic applications such as automotive three-way catalytic systems in which toxic CO and NO are oxidized and reduced to safe CO2 and N2 , respectively. In this work, we explore the incorporation of Cu and Cr metals as dopants in the crystal structure of ceria nanorods prepared by a single-step hydrothermal synthesis. XRD, Raman and XPS confirm the incorporation of Cu and Cr in the ceria crystal lattices, offering ceria nanorods with a higher concentration of oxygen vacancies. XPS also confirms the presence of Cr and Cu surface species. H2 -TPR and XPS analysis show that the simultaneous Cu and Cr co-doping results in a catalyst with a higher surface Cu concentration and a much-enhanced surface reducibility, in comparison with either undoped or singly doped (Cu or Cr) ceria nanorods. While single Cu doping enhances catalytic CO oxidation and Cr doping improves catalytic NO reduction, co-doping with both Cu and Cr enhances the benefits of both dopants in a synergistic manner employing roughly a quarter of dopant weight.


Current Trends in Subspecialty Fellowship Training for 1691 Academic Neurological Surgeons.
Neurosurgical subspecialty fellowship training has become increasingly popular in recent decades. However, few studies have evaluated recent trends in postgraduate subspecialty education. This study aims to provide a detailed cross-sectional analysis of subspecialty fellowship training completion trends and demographics among U.S. academic neurosurgeons. Academic clinical faculty (M.D. or D.O.) teaching at accredited neurosurgery programs were included. Demographic, career, and fellowship data were collected from departmental physician profiles and the American Association of Neurological Surgeons (AANS) membership database. Relative citation ratio scores were retrieved using the National Institutes of Health iCite tool. This study included 1691 surgeons (1756 fellowships) from 125 institutions. The majority (79.13%) reported fellowship training. Fellowship completion was more common among recent graduates (residency year >2000), as was training in multiple subspecialties (P < 0.0001). Spine was the most popular subspecialty (16.04%), followed by pediatrics (11.18%), and cerebrovascular (9.46%). The least common were trauma/critical care (2.52%) and peripheral nerve (1.26%). Spine, neuroradiology, and endovascular subspecialties grew in popularity over time. Pediatrics and spine were the most popular for females and males, respectively. Epilepsy and cerebrovascular had the most full professors, while endovascular and spine had the most assistant professors. Stereotactic/functional and epilepsy had the most Ph.Ds. Fellowship training correlated with higher weighted, but not mean, relative citation ratio scores among associate (P = 0.002) and full professors (P = 0.005). There is an emerging proclivity for additional fellowship training among young neurosurgeons, often in multiple subspecialties. These findings are intended to help guide professional decision-making and optimize the delivery of postgraduate education.


Directed electron regulation promoted sandwich-like CoO@FeBTC/NF with p-n heterojunctions by gel electrodeposition for oxygen evolution reaction.
Metal organic framework (MOF) is currently-one of the key catalysts for oxygen evolution reaction (OER), but its catalytic performance is severely limited by electronic configuration. In this study, cobalt oxide (CoO) on nickel foam (NF) was first prepared, which then wrapped it with FeBTC synthesized by ligating isophthalic acid (BTC) with iron ions by electrodeposition to obtain CoO@FeBTC/NF p-n heterojunction structure. The catalyst requires only 255 mV overpotential to reach a current density of 100 mA cm-2, and can maintain 100 h long time stability at 500 mA cm-2 high current density. The catalytic properties are mainly related to the strong induced modulation of electrons in FeBTC by holes in the p-type CoO, which results in stronger bonding and faster electron transfer between FeBTC and hydroxide. At the same time, the uncoordinated BTC at the solid-liquid interface ionizes acidic radicals which form hydrogen bonds with the hydroxyl radicals in solution, capturing them onto the catalyst surface for the catalytic reaction. In addition, CoO@FeBTC/NF also has strong application prospects in alkaline electrolyzers, which only needs 1.78 V to reach a current density of 1 A cm-2, and it can maintain long-term stability for 12 h at this current. This study provides a new convenient and efficient approach for the control design of the electronic structure of MOF, leading to a more efficient electrocatalytic process.


Evaluation of a commercial enzyme immunoassay for detection of Mycoplasma pneumoniae specific immunoglobulin G antibodies.
A commercial enzyme immunoassay (Platelia Mycoplasma, Diagnostics Pasteur) for the diagnosis of Mycoplasma pneumoniae [corrected] infections was evaluated and found not to be suitable for the purpose. More than 80% of healthy persons and patients with non-Mycoplasma pneumoniae respiratory infection, all with a negative Mycoplasma pneumoniae complement fixation test, had a positive EIA. Paired sera did not show the positive correlation between a rise in complement fixation titre and the EIA ratio reported by the manufacturer.


Alkaline DNA fragmentation, DNA disentanglement evaluated viscosimetrically and sister chromatid exchanges, after treatment in vivo with nitrofurantoin.
Nitrofurantoin was not positive as a carcinogen in long term assays. In vitro it was positive in some short term tests and negative in others. We have examined Nitrofurantoin for its capability of inducing DNA damage in vivo. With the alkaline elution technique, Nitrofurantoin appeared clearly positive in all the tissues examined (liver, kidney, lung, spleen and bone marrow). In the liver we also observed some cross-linking effect. In bone marrow cells Nitrofurantoin was also clearly positive in terms of sister chromatid exchanges (SCEs) induction. DNA damage in vivo was also examined with a viscosimetric method, more sensitive than alkaline elution. With this method the results were essentially negative, suggesting that the two methods detect different types of damage. In view of its positivity in many organs and in two short term tests in vivo, the carcinogenic potential of Nitrofurantoin should be reconsidered.


Vitamin D Endocrine System and COVID-19.
Preclinical data strongly suggest that the vitamin D endocrine system (VDES) may have extraskeletal effects. Cells of the immune and cardiovascular systems and lungs can express the vitamin D receptor, and overall these cells respond in a coherent fashion when exposed to 1,25-dihydroxyvitamin D, the main metabolite of the VDES. Supplementation of vitamin D-deficient subjects may decrease the risk of upper respiratory infections. The VDES also has broad anti-inflammatory and anti-thrombotic effects, and other mechanisms argue for a potential beneficial effect of a good vitamin D status on acute respiratory distress syndrome, a major complication of this SARS-2/COVID-19 infection. Activation of the VDES may thus have beneficial effects on the severity of COVID-19. Meta-analysis of observational data show that a better vitamin D status decreased the requirement of intensive care treatment or decreased mortality. A pilot study in Cordoba indicated that admission to intensive care was drastically reduced by administration of a high dose of calcifediol early after hospital admission for COVID-19. A large observational study in Barcelona confirmed that such therapy significantly decreased the odds ratio (OR) of mortality (OR = 0.52). This was also the conclusion of a retrospective study in five hospitals of Southern Spain. A retrospective study on all Andalusian patients hospitalized because of COVID-19, based on real-world data from the health care system, concluded that prescription of calcifediol (hazard ratio [HR] = 0.67) or vitamin D (HR = 0.75), 15 days before hospital admission decreased mortality within the first month. In conclusion, a good vitamin D status may have beneficial effects on the course of COVID-19. This needs to be confirmed by large, randomized trials, but in the meantime, we recommend (rapid) correction of 25 hydroxyvitamin D (25OHD) deficiency in subjects exposed to this coronavirus. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.


Enhanced quorum sensing of anode biofilm for better sensing linearity and recovery capability of microbial fuel cell toxicity sensor.
MFC toxicity sensor has major hindrances that limit its practical application, such as the poor concentration-response relationship and inferior recovery capability after high toxicity shock. Till now, the direct influence of intrinsic properties on the performance of MFC toxicity sensor has not been well understood. Quorum sensing (QS) is a cell-to-cell communication strategy that indirectly affects the intrinsic properties of electroactive biofilms. In this work, commercially available QS autoinducers (AHLs) were applied to MFC toxicity sensor to manipulate anode biofilm for better sensing performance. The results showed that the addition of AHLs (C6-HSL, 3-OXO-C12-HSL) led to higher sensing linearity to a wider range of Pb2+. The voltage of MFC sensors with AHLs addition fully recovered even after 10 mg/L Cu2+ shock, indicating an enhanced recovery capability of MFC toxicity sensor. It was found that higher live/dead cells ratio and increased exoelectrogen Geobacter abundance were responsible for the superior sensing linearity and recovery capability of MFC toxicity sensor. Our work presented a novel and effective way to advance the process of MFC toxicity sensor application from the perspective of EABs.


Analysis on mechanical properties and mesoscopic acoustic emission characteristics of prefabricated fracture cemented paste backfill under different loading rates.
The mechanical characteristics of cemented paste backfill (CPB) are significantly influenced by the loading rate (LR) and initial defects. Therefore, the CPB with prefabricated fracture (PF, PFCPB) was prepared, and uniaxial compressive strength (UCS) tests considering LR and acoustic emission (AE) monitoring were performed. The particle flow code (PFC2D) was introduced to analyze the mesoscopic crack evolution of the filling body, and the moment tensor theory was used to invert the AE signals characteristics. The results show that as the PF angle increased, the UCS and elastic modulus (EM) of PFCPB decreased and then increased, and the 30° PF was the turning point. The mechanical properties of PFCPB were deteriorated by the presence of PF. Meanwhile, the mechanical properties of PFCPB were positively correlated with the LR. The stress-strain curve of PFCB (excluding 90°) showed bimodal curves. After the UCS test, the macro crack of PFCPB sprouted at the tip of PF or converged toward PF. PFCPB mainly suffered from shear failure, accompanied by a few tensile failures. Numerical simulation results showed that the crack initiation stress of PFCPB was reduced by the PF. The number of cracks first dropped and then gradually increased when the PF angle was enhanced, while gradually increased with the LR increased. Meanwhile, the mesoscopic AE characteristics of CPB were strongly in line with the test results. The AE events of 0 ~ 60° PFCPB experienced two slow rising periods and rapid rising periods. The temporal and spatial distribution characteristics of AE corresponded to the crack evolution trend. The PF was prone to stress concentration, especially at the tip and upper and lower surfaces of 0 ~ 45° PF, resulting in rapid crack initiation and reducing the energy storage limit and mechanical behavior of 0 ~ 45° PFCPB. The increasing LR (within a certain range) was in favor of improving the mechanical behavior of the filling body. The research results can provide a basic reference for the stability evaluation of the filling body with initial defects.


Courses of treatment and risk factors for treatment-resistant depression in Finnish primary and special healthcare: A nationwide cohort study.
Investigate incidence, risk factors and courses of treatment for treatment-resistant depression (TRD) in primary and special healthcare. All patients identified from nationwide registers, aged 16-65 years, diagnosed with depression in Finland during 2004-2016 were included. New antidepressant users were identified with six-month washout period and followed-up for two years to observe for presence of TRD, which was defined as initiation of a third trial after having failed two pharmacological treatment trials with adequate duration. During follow-up, 177,144 persons had their first registered antidepressant treated depression (mean age: 39.5, 62.5% women). Of them, 10.9% (N = 19,322) met TRD criteria. Among the TRD patients, most common first and second antidepressants trials were: SSRIs (44.6%), mirtazapine (19.0%) and SNRIs (16.5%). As the third treatment line, antidepressant monotherapy (44.2% of TRD patients) was most common, followed by a combination of ≥2 antidepressants (32.1%), antipsychotic or mood stabilizer augmentation and an antidepressant (15.8%), both combination of antidepressants and an augmentation with a mood stabilizer or antipsychotic (4.9%), antipsychotic or mood stabilizer monotherapy (2.7%) and ECT (0.3%). Of TRD patients, 16.5% (N = 3188) progressed to the fifth treatment line, in which the most common treatments were antidepressant monotherapy (33.4%), antidepressant combinations (27.5%) and augmentation (24.2%). Factors associated with higher risk of TRD included male gender, younger age, higher initial disease severity and hospitalization at initial onset of depression. Antidepressant monotherapies were still the most common fifth line of depression treatment. Severe depression, hospitalization due to depression, young age and male gender may predispose to TRD.


Retracted: Feature Sequencing Method of Industrial Control Data Set Based on Multidimensional Evaluation Parameters.
[This retracts the article DOI: 10.1155/2022/9248267.].


[Ultrasound-guided peripheral venous access].
International guidelines advocate the use of first-line ultrasound for central venous catheter, particularly for the internal jugular vein. The role of ultrasound in peripheral venous access remains questionable. In some specific situations, such as pediatrics, obesity and patients with poor venous network, problems to cannulate peripheral vein may occur. Success rate of peripheral intravenous access increases with the diameter of the vein and for a depth of the vein between 0.3 and 1.5 cm. The type of puncture (long-axis or short-axis) and the type of catheters have little influence on the success rate. Specific considerations have to be taken concerning infection control.


Relatives' Presence During Cardiopulmonary Resuscitation.
The question of whether or not to allow family to be present during resuscitation is relevant to everyday professional health care assistance, but it remains largely unexplored in the medical literature. We conducted an online survey with the aim of increasing our knowledge and understanding of this issue. This is a cross-sectional, multicenter, descriptive, national, and international study using a web-based, voluntary survey. The survey was designed and distributed through a medical website in Spanish, targeting physicians who frequently deal with critical patients. A total of 1,286 Argentine physicians and 1,848 physicians from other countries responded to this voluntary survey. Of Argentine respondents, 15.8% (203) treat only children, 68.2% (877) treat adults, and 16% (206) treat patients of any age. The survey found that 23% (296) of Argentine and 20% of other respondents favor the presence of family members during cardiopulmonary resuscitation (p = 0.03). This practice was more common among physicians treating pediatric and neonatal patients than among those who treat adults. The most commonly reported reason (21.8%) for avoiding the presence of relatives was concerns that physicians, communications, and medical practices might be misunderstood or misinterpreted. Avoiding relatives' presence while performing cardiopulmonary resuscitation is the most frequent choice made by the surveyed physicians who treat critical Argentine patients. The main causes for discouraging family presence during cardiopulmonary resuscitation or other critical procedures include the following: risk of misinterpretation of the physician's actions and/or words; risk of a relative's decompensation; uncertainty about possible reactions; and interpretation of the relative's presence as negative.


Development of a novel fusion imaging technique in the diagnosis of hepatobiliary-pancreatic lesions.
Multi-row detector computed tomography (MDCT) and magnetic resonance cholangiopancreatography (MRCP) play an important role in the imaging diagnosis of hepatobiliary-pancreatic lesions. Here we investigated whether unifying the MDCT and MRCP images onto the same screen using fusion imaging could overcome the limitations of each technique, while still maintaining their benefits. Moreover, because reports of fusion imaging using MDCT and MRCP are rare, we assessed the benefits and limitations of this method for its potential application in a clinical setting. The patient group included 9 men and 11 women. Among the 20 patients, the final diagnoses were as follows: 10 intraductal papillary mucinous neoplasms, 5 biliary system carcinomas, 1 pancreatic adenocarcinoma and 5 non-neoplastic lesions. After transmitting the Digital Imaging and Communication in Medicine data of the MDCT and MRCP images to a workstation, we performed a 3-D organisation of both sets of images using volume rendering for the image fusion. Fusion imaging enabled clear identification of the spatial relationship between a hepatobiliary-pancreatic lesion and the solid viscera and/or vessels. Further, this method facilitated the determination of the relationship between the anatomical position of the lesion and its surroundings more easily than either MDCT or MRCP alone. Fusion imaging is an easy technique to perform and may be a useful tool for planning treatment strategies and for examining pathological changes in hepatobiliary-pancreatic lesions. Additionally, the ease of obtaining the 3-D images suggests the possibility of using these images to plan intervention strategies.


Candida sonckii sp. nov.
A new yeast species with basidiomycetous affinities is described as Candida sonckii Hopsu-Havu et al. It differs from C. fujisanensis and C. maris by growing with KNO3 as nitrogen source, from C. vanderwaltii by its lower mol% G + C, lower maximum temperature for growth and failure to grow on D-arabinose and citrate.


Insulin secretion and hepatic extraction in humans by minimal modeling of C-peptide and insulin kinetics.
Methods for measuring insulin secretion and hepatic insulin extraction in vivo, e.g., hepatic vein catheterization, are invasive, and can be applied during steady state only. We introduce a noninvasive method for measuring in vivo insulin secretion and its extraction by the liver during an intravenous glucose tolerance test (IVGTT). This method is based on a minimal model of C-peptide secretion and kinetics that is used for interpreting plasma C-peptide concentration data during an IVGTT in normal humans. The model allows the reconstruction of the time course of insulin secretion and, used in conjunction with a minimal model of insulin delivery and kinetics (described in a previous study), provides a noninvasive measure of the time course of hepatic insulin extraction [H(t)]. The C-peptide model also provides a direct prehepatic measure of beta-cell sensitivity to glucose, expressed by two parameters related to first (phi IC)- and second (phi IIC)-phase insulin secretion. In the 11 healthy volunteers we studied, these parameters were 61 +/- 11 pM.min-1.mg-1.dl and 0.0154 +/- 0.0034 pM.min-2.mg-1.dl, respectively. H(t) showed an initial decrement for approximately 30-50 min (from a fasting value of 63 +/- 8% to a nadir of 53 +/- 9%) after the glucose stimulus, then a steady value of approximately 62% was reestablished and maintained throughout the experiment. The validity of the C-peptide model was further assessed by comparing its estimate of the fractional plasma clearance rate (k01) with that obtained in experiments in which biosynthetic human C-peptide was administered.(ABSTRACT TRUNCATED AT 250 WORDS)


[Idiopathic severe valgus deformity of the knee in children. Apropos of 2 cases].
The authors have reviewed two cases of important and idiopathic deformity in children coming from Africa. In the third year of life, a valgus deformity of a knee occurs spontaneously and increases rapidly in a previously healthy child. Varus deformity of the other knee follows but seems to be just a mechanical consequence of the valgus deformity. The pathological lesions observed radiologically concern mainly the metaphyseses. General skeletal or metabolic disease, malnutrition or sickle-cell disease are not found.


Where health and environment meet: the use of invariant parameters in big data analysis.
The use of big data to investigate the spread of infectious diseases or the impact of the built environment on human wellbeing goes beyond the realm of traditional approaches to epidemiology, and includes a large variety of data objects produced by research communities with different methods and goals. This paper addresses the conditions under which researchers link, search and interpret such diverse data by focusing on "data mash-ups"-that is the linking of data from epidemiology, biomedicine, climate and environmental science, which is typically achieved by holding one or more basic parameters, such as geolocation, as invariant. We argue that this strategy works best when epidemiologists interpret localisation procedures through an idiographic perspective that recognises their context-dependence and supports a critical evaluation of the epistemic value of geolocation data whenever they are used for new research purposes. Approaching invariants as strategic constructs can foster data linkage and re-use, and support carefully-targeted predictions in ways that can meaningfully inform public health. At the same time, it explicitly signals the limitations in the scope and applicability of the original datasets incorporated into big data collections, and thus the situated nature of data linkage exercises and their predictive power.


Long-term results after aortic valve replacement with the Bravo 400 stentless xenograft.
This study was undertaken to evaluate the long-term clinical and echocardiographic outcome after aortic valve replacement with the Bravo Cardiovascular Model 400 stentless xenograft. Between February 1992 and January 1994, 67 patients underwent aortic valve replacement with the Bravo 400 bioprosthesis. The valvular pathology was aortic stenosis in 36 patients (53.7%), aortic insufficiency in 17 patients (25.4%), and mixed lesion in 14 patients (20.9%). Mean follow-up time was 9.8 +/- 2.73 years and median follow-up time was 11 years. Cumulative follow-up time was 659 patients-years and was 94% complete. No early deaths were observed. Overall survival estimates at 11 years were 74.71% +/- 5.47%. The actuarial freedom from valve-related death at 11 years was 91.04% +/- 3.84%; from cardiac-related death at 11 years it was 87.95% +/- 4.29%; and from noncardiac death at 11 years it was 85.14% +/- 4.58%. Eleven-year Kaplan-Meier survival of patients younger than 65 years was 90.91% +/- 6.13% versus 66.08% +/- 7.38% for older patients (p = 0.0307, log-rank test). The actuarial freedom from all valve-related morbidity and mortality at 11 years was 80.3% +/- 5.4%. The mean transvalvular gradient decreased significantly after aortic valve replacement with a corresponding increase in effective orifice area. Left ventricular mass index at 10-year follow-up was 68.5% of the preoperative value. The Bravo Cardiovascular Model 400 stentless xenograft has provided good clinical and hemodynamic results up until 11 years of follow-up.


Disruption of dTDP-rhamnose biosynthesis modifies lipopolysaccharide core, exopolysaccharide production, and root colonization in Azospirillum brasilense.
The interaction between Azospirillum brasilense and plants is not fully understood, although several bacterial surface components like exopolysaccharides (EPS), flagella, and capsular polysaccharides are required for attachment and colonization. While in other plant-bacteria associations (Rhizobium-legume, Pseudomonas-potato), lipopolysaccharides (LPS) play a key role in the establishment of an effective association, their role in the root colonization by Azospirillum had not been determined. In this study, we isolated a Tn5 mutant of A. brasilense Cd (EJ1) with an apparently modified LPS core structure, non-mucoid colony morphology, increased EPS production, and affected in maize root colonization. A 3790-bp region revealed the presence of three complete open reading frames designated rmlC, rmlB and rmlD. The beginning of a fourth open reading frame was found and designated rmlA. These genes are organized in a cluster which shows homology to the cluster involved in the synthesis of dTDP-rhamnose in other bacteria. Additionally, the analysis of the monosaccharide composition of LPSs showed a diminution of rhamnose compared to the wild-type strain.


Balance and eye movement training to improve gait in people with progressive supranuclear palsy: quasi-randomized clinical trial.
Although vertical gaze palsy and gait instability are cardinal features of progressive supranuclear palsy (PSP), little research has been done to address oculomotor and gait rehabilitation for PSP. The purpose of this study was to compare the benefits of a program of balance training complemented with eye movement and visual awareness training versus balance training alone to rehabilitate gait in people with PSP. Nineteen people moderately affected by the disease were assigned to either a treatment group (balance plus eye movement exercises, n=10) or a comparison group (balance exercises only, n=9) in a quasi-random fashion. The baseline characteristics assessed were diagnosis (possible versus probable), sex, age, time of symptom onset, dementia, and severity of symptoms. Within-group, between-group, and effect size analyses were performed on kinematic gait parameters (stance time, swing time, and step length) and clinical tests (8-ft [2.4-m] walk test and Timed "Up & Go" Test). The within-group analysis revealed significant improvements in stance time and walking speed for the treatment group, whereas the comparison group showed improvements in step length only. Moderate to large effects of the intervention were observed for the treatment group, and small effects were observed for the comparison group. The between-group analysis did not reveal significant changes for either group. These preliminary findings support the use of eye movement exercises as a complementary therapy for balance training in the rehabilitation of gait in people with PSP and moderate impairments. Additional studies powered at a higher level are needed to confirm these results.


Effects of Atractylodes macrocephala on the cytomembrane Ca2+-activated K+ currents in cells of human pregnant myometrial smooth muscles.
The study examined the inhibitory effect of Atractylodes macrocephala (AM) on the uterine contraction during premature delivery and explored its electrophysiological mechanism by studying the effects of AM on the Ca(2+)-activated K(+) currents of pregnant human myometrial smooth muscle cells with or without the treatment with interleukin-6. Single cells were acutely isolated from pregnant human myometrial smooth muscles. Whole-cell Ca(2+)-activated K(+) currents were recorded by using an Axopatch1-D amplifier. The cells were divided into three groups: group A in which AM was added into perfusate, group B, in which interleukin-6 was added into perfusate) and group C in which AM was added into perfusate after addition of interleukin-6. IL-6 10 ng/mL inhibited BK(ca) by 36.9%+/-13.7% as compared with control (P<0.01). AM at 2 mg/mL raised BK(ca) by 36.7%+/-22.6% or 45.2%+/-13.7% with or without the treatment of IL-6, respectively (P<0.01). It is concluded that AM was able to enhance the BK(ca) of pregnant human myometrial smooth muscle cells treated or untreated with interleukin-6 and its effect on the BK(ca) IL-treated cells was stronger that its effect on BK(ca) of untreated cells. Our results suggested that AM can help to maintain the membrane potentials and the resting status of pregnant human myometrial smooth muscle cells.


The contractile system as a negative regulator of the connexin 43 hemichannel.
The molecular mechanisms underlying the regulation of gap junction (GJ) channels based on the 43-kDa connexin isoform (Cx43) have been studied extensively. GJ channels are formed by the docking of opposed hemichannels in adjacent cells. Mounting data indicate that unopposed Cx43 hemichannels are also functional in the plasma membrane. However, our understanding of how Cx43-hemichannel opening and closing is regulated at the molecular level is only poorly understood. Recent work elucidated that actomyosin contractility inhibits potently Cx43 hemichannels. It is known that intracellular Ca²⁺ exerts a bell-shaped-dependent effect on Cx43-hemichannel opening. While low-intracellular [Ca²⁺] (<500 nM) provokes opening of the channel, high-intracellular [Ca²⁺] (> 500 nM) favours closing of the channel. The mechanism underlying this negative regulation of Cx43-hemichannel activity by high-intracellular [Ca²⁺] seems to be dependent on the activation of the actomyosin contractile system. The activity of Cx43 hemichannels is critically controlled by molecular interactions between the intracellular loop and the C-terminal tail. These interactions are essential for Cx43-hemichannel opening in response to triggers such as cytosolic [Ca²⁺] rise or external [Ca²⁺] lowering. In this review, we present the hypothesis that the actomyosin contractile system can function as an important brake mechanism on Cx43-hemichannel opening. By controlling loop-tail interactions, the contractile system would prevent aberrant or excessive opening of Cx43 hemichannels.


The purinergic P2Y14 receptor links hepatocyte death to hepatic stellate cell activation and fibrogenesis in the liver.
Fibrosis contributes to ~45% of deaths in western countries. In chronic liver disease, fibrosis is a major factor determining outcomes, but efficient antifibrotic therapies are lacking. Although platelet-derived growth factor and transforming growth factor-β constitute key fibrogenic mediators, they do not account for the well-established link between cell death and fibrosis in the liver. Here, we hypothesized that damage-associated molecular patterns (DAMPs) may link epithelial cell death to fibrogenesis in the injured liver. DAMP receptor screening identified purinergic receptor P2Y14 among several candidates as highly enriched in hepatic stellate cells (HSCs), the main fibrogenic cell type of the liver. Conversely, P2Y14 ligands uridine 5'-diphosphate (UDP)-glucose and UDP-galactose were enriched in hepatocytes and were released upon different modes of cell death. Accordingly, ligand-receptor interaction analysis that combined proteomic and single-cell RNA sequencing data revealed P2Y14 ligands and P2Y14 receptor as a link between dying cells and HSCs, respectively. Treatment with P2Y14 ligands or coculture with dying hepatocytes promoted HSC activation in a P2Y14-dependent manner. P2Y14 ligands activated extracellular signal-regulated kinase (ERK) and Yes-associated protein (YAP) signaling in HSCs, resulting in ERK-dependent HSC activation. Global and HSC-selective P2Y14 deficiency attenuated liver fibrosis in multiple mouse models of liver injury. Functional expression of P2Y14 was confirmed in healthy and diseased human liver and human HSCs. In conclusion, P2Y14 ligands and their receptor constitute a profibrogenic DAMP pathway that directly links cell death to fibrogenesis.


Graphene oxide-based magnetic fluorescent hybrids for drug delivery and cellular imaging.
A graphene oxide (GO)-based multifunctional hybrid has been developed for loading and delivery of anticancer drugs. The GO was functionalized by magnetic/fluorescent SiO2 microsphere via an amidation process. Doxorubicin (DOX) was chosen as a model drug to be loaded onto the GO via π-π stacking and hydrophobic interaction. The loading capacities of DOX to the GO-based magnetic fluorescent hybrids were investigated. The release profiles of DOX from the hybrids were depicted. The fluorescence images of the GO-based magnetic fluorescent hybrids indicated that the hybrids would be an effective drug carrier and a potential optical imaging tool. The application of the developed GO-based magnetic fluorescent hybrids for HepG2 cells demonstrated an excellent therapeutic effect of the DOX loaded hybrids to the target tumor.


Efficient electrotransformation of Enterococcus hirae with a new Enterococcus-Escherichia coli shuttle vector.
In the present study, an Enterococcus-Escherichia coli shuttle vector, pC3, was constructed that allows efficient transformation by electroporation of Enterococcus hirae ATCC9790. 5 x 10(6) transformants per microgram of plasmid DNA were obtained, using a commercial capacitor discharge device with an improved circuitry and a home-made electrode assembly, delivering pulses of 24 kV/cm across the cell suspension. The transformants were stable without selective pressure and plasmid DNA reisolated from transformed cells displayed no alterations in restriction enzyme analysis. Chromosomal DNA from E coli or E hirae, carried by pC3, was stably maintained in E hirae, making cloning and genetic manipulation in this organism feasible.


Prediction by (13)C NMR of regioselectivity in 1,3-dipolar cycloadditions of acridin-9-yl dipolarophiles.
Strong correlation was found between (13)C NMR chemical shifts of dipolarophilic CH=CH carbons and regioselectivity in 1,3-dipolar cycloadditions of new acridin-9-yl dipolarophiles with stable benzonitrile oxides (BNO). Accordingly, two starting dipolarophiles, (acridin-9-yl)-CH=CH-R (R = COOCH3 or Ph), reacted with three BNOs (2,4,6-trimethoxy, 2,4,6-trimethyl, and 2,6-dichloro) to give a mixture of two target isoxazoline regioisomers in which the acridine was bound either to isoxazoline C-4 carbon (4-Acr) or C-5 one (5-Acr). Methyl 3-(acridin-9-yl)propenoate afforded major 4-(acridin-9-yl)-isoxazoline-5-carboxylates (4-Acr) and minor 5-(acridin-9-yl)-4-carboxylates (5-Acr). 9-(2-Styryl)acridine regiospecifically afforded only 4-Acr cycloadducts. The ratios of regioisomers were compared with analogous reactions of acridin-4-yl dipolarophiles. Regioselectivity was dependent on a polarity of the CH=CH bond, donor effects in BNO, and stabilization by stacking of aromatic substituents in the products.


Reactivity of nitrogen species with inorganic and organic compounds in water.
Many studies on the reactive nitrogen species (RNS, ●NO2, ●NO and ●NH2) with pollutants in water have been performed to understand the abatement of inorganic and organic compounds by these species, and the mechanisms of the formation of oxidative transformation products, especially nitrogenous oxidized byproducts. In this review, approaches to generate RNS in aqueous solution is first presented, followed by a summary of their reactivity with a wide range of compounds. The second-order rate constants (k, M-1 s-1) for the reactivity of ●NO2 and ●NO with a wide range of inorganic radical and nonradical species were correlated with thermodynamic one-electron oxidation potentials (E0). The positive correlation between log(k) versus E0 suggests one-electron transfer reactions. The Hammett-type correlations were developed for the reactions of ●NO2 and ●NH2 with organic compounds, using the unsubstituted benzene as a reference molecule (i.e., Σσo,p,m = 0) to calculate Σσo,p,m = σo + σp + σm for each organic molecule. Linear negative correlations of log(k) with Σσo,p,m were obtained for both ●NO2 and ●NH2, suggesting electrophilic substitution mechanism. The correlations presented herein may assist in eliminating organic micropollutants in water treatment and reuse processes.


Effect of treatment medium on induction of aneuploidy by nocodazole in Saccharomyces cerevisiae.
While studying ways to improve responsiveness of Saccharomyces cerevisiae strain D61.M to agents that induce aneuploidy, we noted that nocodazole, which strongly induces aneuploidy when yeast cells are treated in yeast extract-peptone-dextrose (YEPD) medium, had no effect when a synthetic complete (SC) medium was used. Further study revealed that the presence of peptone was necessary for induction. Other aneuploidy-inducing agents, including ethyl acetate, acetone, and methyl benzimidazole-2-yl-carbamate (MBC), were equally active in either medium. Benomyl, which degrades to MBC, was less active in SC than in YEPD medium.


Evidence for an indirect cholinergic regulation of blood flow in the hypothalamus of conscious rabbits.
1 The effects of methacholine, atropine and adrenoceptor blockade on hypothalamic blood flow (HBF) were measured in conscious rabbits. 2 A dose of 1 microgram methacholine increased HBF while smaller and larger doses had no significant effect. 3 The vasodilatation induced by methacholine was blocked by atropine and by chemical sympathectomy of the hypothalamus with 6-hydroxydopamine. 4 The vasodilatation was reversed by propranolol but was not affected by phenoxybenzamine. 5 These results suggest that the vasodilator action of muscarinic receptor agonists on hypothalamic resistance vessels depends upon the integrity of a noradrenergic system, and is mediated via beta-adrenoceptors.


RNAi-Mediated Silencing of Catalase Gene Promotes Apoptosis and Impairs Proliferation of Bovine Granulosa Cells under Heat Stress.
Heat stress in dairy cattle is recognized to compromise fertility by altering the functions of ovarian follicle-enclosed cells, e.g., oocyte and granulosa cells (GCs). Catalase is an antioxidant enzyme that plays a significant role in cellular protection against oxidative damage by the degradation of hydrogen peroxide to oxygen and water. In this study, the role and mechanism of CAT on the heat stress (HS)-induced apoptosis and altered proliferation of bovine GCs were studied. The catalase gene was knocked-down successfully in bovine GCs at both the transcriptional and translational levels. After a successful knockdown using siRNA, GCs were divided into HS (40 °C + NC and 40 °C + CAT siRNA) and 38 °C + NC (NC) groups. The GCs were then examined for ROS, viability, mitochondrial membrane potential (MMP), cell cycle, and biosynthesis of progesterone (P4) and estrogen (E2) hormones. The results indicated that CAT silencing promoted ROS production and apoptosis by up-regulating the Bcl-2-associated X protein (BAX) and Caspase-3 genes both at the transcriptional and translational levels. Furthermore, the knockdown of CAT markedly disrupted the MMP, impaired the production of P4 and E2, altered the progression of the G1 phase of the cell cycle, and decreased the number of cells in the S phase. This was further verified by the down-regulation of proliferating cell nuclear antigen (PCNA), CyclinB1, steroidogenic acute regulatory protein (STAR), and cytochrome P450 family 11 subfamily A member 1 (Cyp11A1) genes. Our study presented a novel strategy to characterize how CAT can regulate cell proliferation and apoptosis in GCs under HS. We concluded that CAT is a broad regulatory marker in GCs by regulating apoptosis, cellular progression, and simultaneously by vital fluctuations in hormonal signaling. Our findings infer a crucial evidence of how to boost the fertility of heat-stressed cows.


Activation of autophagy during normothermic machine perfusion of discarded livers is associated with improved hepatocellular function.
Liver transplantation is hampered by a severe shortage of donor organs. Normothermic machine perfusion (NMP) of donor livers allows dynamic preservation in addition to viability assessment before transplantation. Little is known about the injury and repair mechanisms induced during NMP. To investigate these mechanisms, we examined gene and protein expression changes in a cohort of discarded human livers, stratified by hepatocellular function, during NMP. Six human livers acquired through donation after circulatory death (DCD) underwent 12 h of NMP. Of the six livers, three met predefined criteria for adequate hepatocellular function. We applied transcriptomic profiling and protein analysis to evaluate temporal changes in gene expression during NMP between functional and nonfunctional livers. Principal component analysis segregated the two groups and distinguished the various perfusion time points. Transcriptomic analysis of biopsies from functional livers indicated robust activation of innate immunity after 3 h of NMP followed by enrichment of prorepair and prosurvival mechanisms. Nonfunctional livers demonstrated delayed and persistent enrichment of markers of innate immunity. Functional livers demonstrated effective induction of autophagy, a cellular repair and homeostasis pathway, in contrast to nonfunctional livers. In conclusion, NMP of discarded DCD human livers results in innate immune-mediated injury, while also activating autophagy, a presumed mechanism for support of cellular repair. More pronounced activation of autophagy was seen in livers that demonstrated adequate hepatocellular function.NEW & NOTEWORTHY We demonstrate that ischemia-reperfusion injury occurs in all livers during NMP, though there are notable differences in gene expression between functional and nonfunctional livers. We further demonstrate that activation of the liver's repair and homeostasis mechanisms through autophagy plays a vital role in the graft's response to injury and may impact liver function. These findings indicate that liver autophagy might be a key therapeutic target for rehabilitating the function of severely injured or untransplantable livers.