miR-204-5p Represses Bone Metastasis via Inactivating NF-κB Signaling in Prostate Cancer.
The prime issue derived from prostate cancer (PCa) is its high prevalence to metastasize to bone. MicroRNA-204-5p (miR-204-5p) has been reported to be involved in the development and metastasis in a variety of cancers. However, the clinical significance and biological functions of miR-204-5p in bone metastasis of PCa are still not reported yet. In this study, we find that miR-204-5p expression is reduced in PCa tissues and serum sample with bone metastasis compared with that in PCa tissues and serum sample without bone metastasis, which is associated with advanced clinicopathological characteristics and poor bone metastasis-free survival in PCa patients. Moreover, upregulation of miR-204-5p inhibits the migration and invasion of PCa cells in vitro, and importantly, upregulating miR-204-5p represses bone metastasis of PCa cells in vivo. Our results further demonstrated that miR-204-5p suppresses invasion, migration, and bone metastasis of PCa cells via inactivating nuclear factor κB (NF-κB) signaling by simultaneously targeting TRAF1, TAB3, and MAP3K3. In clinical PCa samples, miR-204-5p expression negatively correlates with TRAF1, TAB3, and MAP3K3 expression and NF-κB signaling activity. Therefore, our findings reveal a new mechanism underpinning the bone metastasis of PCa, as well as provide evidence that miR-204-5p might serve as a novel serum biomarker in bone metastasis of PCa. This study identifies a novel functional role of miR-204-5p in bone metastasis of prostate cancer and supports the potential clinical value of miR-204-5p as a serum biomarker in bone metastasis of PCa.


Utilization of maternal and embryonic histone RNA in early sea urchin development.
Histone RNA in early sea urchin embryos is derived from maternal stores and from new transcription. We show that the sedimentation of maternal free RNPs, containing histone RNA, is somewhat more rapid than the sedimentation of the newly made histone RNPs. Yet, prior to the 2- to 4-cell stage, both the maternally derived and the newly synthesized histone RNA are localized to the same extent in the non-polysomal-free RNPs, and the timing of their recruitment into embryonic polysomes appears to be the same. The levels of hybridization of histone probe to RNAs in cleaving embryos increases severalfold in intensity, and the increase occurs primarily in the polysomes. These data suggest that new transcription may provide an important contribution to the total histone RNA mass by as early as the 32- to 64-cell stage of development.


Chemical genetic-mediated spatial regulation of protein expression in neurons reveals an axonal function for wld(s).
The degeneration of axons is the underlying pathological process of several neurological disorders. The Wallerian degeneration (Wld(S)) slow protein, which is primarily nuclear, markedly inhibits axonal degeneration. Contradictory models have been proposed to explain its mechanism, including a role in the nucleus, where it affects gene transcription, and roles outside the nucleus, where it regulates unknown effectors. To determine which pool of Wld(S) accounts for its axon-protective effects, we developed a strategy to control the spatial expression of proteins within neurons. This strategy couples a chemical genetic method to control protein stability with microfluidic culturing. Using neurons that are selectively deficient in Wld(S) in axons, we show that the axonal pool of Wld(S) is necessary for protection from axon degeneration. These results implicate an axonal pathway regulated by Wld(S) that controls axon degeneration.


Modification of a dihydropyrrolopyrimidine phosphoinositide 3-kinase (PI3K) inhibitor to improve oral bioavailability.
Phosphoinositide 3-kinase (PI3K) is activated in various human cancer cells and well known as a cancer therapy target. We previously reported a dihydropyrrolopyrimidine derivative as a highly potent PI3K inhibitor that has strong tumor growth inhibition in a xenograft model. In this report, we describe further optimization to improve its bioavailability.


High-performance liquid chromatographic determination of hippuric acid in human urine.
A method is described for the determination of urinary hippuric acid by high-performance liquid chromatography. The method used ethyl acetate extraction for partial clean up of the urine. The separation was carried out on a reversed-phase column using 20% methanol in 0.01 M aqueous potassium phosphate containing 0.5% acetic acid as a mobile phase. The column effluent was monitored with a UV detector at 254 nm. Hippuric acid was separated from other normal urine constituents in less than 10 min. Metabolites of xylene and styrene did not interfere with the assay. Analytical recoveries from urine were excellent and peak height and concentration were linearly related.


Serotonergic innervation of the forebrain: effect of lesions on serotonin and tryptophan hydroxylase levels.
Concentrations of serotonin and tryptophan hydroxylase activity in individual hypothalamic and limbic nuclei, as well as cortical regions, were measured by radioenzymatic microassays after dorsal raphe nucleus lesions and various surgical transections of the brain stem. Fourteen days after lesioning of the dorsal raphe nucleus there were 43-65% decreases in 5-HT concentrations of certain hypothalamic and limbic nuclei. More pronounced decreases were found in the parietal cortex and in the locus coeruleus and substantia nigra. Degenerations of nerve terminals in various regions of the hypothalamus and the limbic system were detected by electron microscopic studies 1-8 days after dorsal raphe nucleus lesions. There was no change in the 5-HT concentration of the spinal cord following dorsal raphe nucleus lesions. Surgical transections at the level of mesencephalic-pontine junction caused no significant changes in 5-HT concentrations or tryptophan hydroxylase levels in the hypothalamus, limbic system, cerebral cortex or spinal cord. Serotonergic fibers ascending from the mesencephalic raphe nuclei constitute a well circumscribed bundle dorsal to the interpeduncular nucleus. The axons enter the hypothalamus between the mammillary body and crus cerebri just ventral to the medial forebrain bundle. The ascending pathway contains all the ascending 5-HT fibers innervating the hypothalamus and the rostral limbic and cortical areas.


Senescent adipocytes as potential effectors of muscle cells dysfunction: An in vitro model.
Recently, there has been a growing body of evidence showing a negative effect of the white adipose tissue (WAT) dysfunction on the skeletal muscle function and quality. However, little is known about the effects of senescent adipocytes on muscle cells. Therefore, to explore potential mechanisms involved in age-related loss of muscle mass and function, we performed an in vitro experiment using conditioned medium obtained from cultures of mature and aged 3 T3-L1 adipocytes, as well as from cultures of dysfunctional adipocytes exposed to oxidative stress or high insulin doses, to treat C2C12 myocytes. The results from morphological measures indicated a significant decrease in diameter and fusion index of myotubes after treatment with medium of aged or stressed adipocytes. Aged and stressed adipocytes presented different morphological characteristics as well as a different gene expression profile of proinflammatory cytokines and ROS production. In myocytes treated with different adipocytes' conditioned media, we demonstrated a significant reduction of gene expression of myogenic differentiation markers as well as a significant increase of genes involved in atrophy. Finally, a significant reduction in protein synthesis as well as a significant increase of myostatin was found in muscle cells treated with medium of aged or stressed adipocytes compared to controls. In conclusion, these preliminary results suggest that aged adipocytes could influence negatively trophism, function and regenerative capacity of myocytes by a paracrine network of signaling.


Thermal transformation of bioactive caffeic acid on fumed silica seen by UV-Vis spectroscopy, thermogravimetric analysis, temperature programmed desorption mass spectrometry and quantum chemical methods.
Thermochemical studies of hydroxycinnamic acid derivatives and their surface complexes are important for the pharmaceutical industry, medicine and for the development of technologies of heterogeneous biomass pyrolysis. In this study, structural and thermal transformations of caffeic acid complexes on silica surfaces were studied by UV-Vis spectroscopy, thermogravimetric analysis, temperature programmed desorption mass spectrometry (TPD MS) and quantum chemical methods. Two types of caffeic acid surface complexes are found to form through phenolic or carboxyl groups. The kinetic parameters of the chemical reactions of caffeic acid on silica surface are calculated. The mechanisms of thermal transformations of the caffeic chemisorbed surface complexes are proposed. Thermal decomposition of caffeic acid complex chemisorbed through grafted ester group proceeds via three parallel reactions, producing ketene, vinyl and acetylene derivatives of 1,2-dihydroxybenzene. Immobilization of phenolic acids on the silica surface improves greatly their thermal stability.


[A nationwide retrospective analysis on chronic diabetic complications and related macrovascular diseases of in-patients with diabetes during 1991-2000].
To investigate the prevalence rates of chronic diabetic complications and their risk factors. We analyzed retrospectively the clinical and laboratory data on the 24,496 in-patients from 30 provinces, municipalities, and autonomous regions during January 1, 1991 to December 31, 2000. The prevalence rates of chronic diabetic complications and related macrovascular diseases were 31.9% for hypertension, 12.2% for cerebrovascular diseases, 15.9% for cardiovascular diseases, 5.0% for lower extremity vascular diseases, 34.3% for retinopathy, 33.6% for nephropathy, and 60.3% for neuropathy, total rate of chronic diabetic complications and macrovascular diseases was 73.2%. The risk factors related to all kinds of chronic diabetic complications and macrovascular diseases were age, illness duration, and systolic blood pressure. The prevalence rates of chronic diabetic complications of in-patients with diabetes mellitus are rather high, near to their level in developed countries. The controllable risk factor of chronic diabetic complications is systolic blood pressure.


Constant density and stable territoriality in some tropical insectivorous birds.
Four species of understory antbirds (Formicariidae: Myrmotherula fulviventris, M. axillaris, Microrhopias quixensis, and Thamnophilus punctatus) had stable populations over eight rainy seasons on Barro Colorado Island, Panama. The co-defended territories of M. fulviventris and Microrhopias quixensis, were essentially identical from year to year on our intensive study site, despite a moderate turnover of territory owners. The location of the territories of T. punctatus was also similar between years. This stability occurred in the face of considerable annual variation in the survivorship of adult M. fulviventris and T. punctatus. This variation was not significantly correlated with patterns of rainfall. Stable territoriality has rarely been reported from relatively-short-lived insectivorous birds. The annual production of young was significantly variable only in M. axillaris. Because BCI is an island comprised of one habitat (tropical forest) and so supports a closed population of antbirds, and because it is unlikely that natality equaled mortality on our study site during the entire eight years of the study, we suggest that these breeding populations are socially regulated at a constant level below the limits directly set by food supply.


Weight discrepancy and body appreciation among women in Poland and Britain.
Previous studies have suggested that the process of transmigration has detrimental effects on the body image of migrants relative to women in the country of origin. In the present work, we examined the body image of Polish migrants in Britain (n=153), Polish women in Poland (n=153), and a comparison group of British White women (n=110). Participants completed a measure of actual-ideal weight discrepancy and the Body Appreciation Scale (BAS). Contrary to hypotheses, our results showed that Polish women in Poland had significantly higher weight discrepancy than their counterparts in Britain. Further analyses showed that the BAS reduced to two dimensions among Polish participants, with Polish participants in Poland having significantly lower body appreciation than Polish migrants. We suggest that the sociocultural changes that have taken place in Eastern Europe may place women in that region at relatively high risk for developing negative body image.


Severe psychosocial deprivation in early childhood is associated with increased DNA methylation across a region spanning the transcription start site of CYP2E1.
Exposure to adverse rearing environments including institutional deprivation and severe childhood abuse is associated with an increased risk for mental and physical health problems across the lifespan. Although the mechanisms mediating these effects are not known, recent work in rodent models suggests that epigenetic processes may be involved. We studied the impact of severe early-life adversity on epigenetic variation in a sample of adolescents adopted from the severely depriving orphanages of the Romanian communist era in the 1980s. We quantified buccal cell DNA methylation at ~400 000 sites across the genome in Romanian adoptees exposed to either extended (6-43 months; n=16) or limited duration (<6 months; n=17) of severe early-life deprivation, in addition to a matched sample of UK adoptees (n=16) not exposed to severe deprivation. Although no probe-wise differences remained significant after controlling for the number of probes tested, we identified an exposure-associated differentially methylated region (DMR) spanning nine sequential CpG sites in the promoter-regulatory region of the cytochrome P450 2E1 gene (CYP2E1) on chromosome 10 (corrected P=2.98 × 10(-5)). Elevated DNA methylation across this region was also associated with deprivation-related clinical markers of impaired social cognition. Our data suggest that environmental insults of sufficient biological impact during early development are associated with long-lasting epigenetic changes, potentially reflecting a biological mechanism linking the effects of early-life adversity to cognitive and neurobiological phenotypes.


Expression and characterization of a functional human immunodeficiency virus envelope glycoprotein in insect cells.
Recombinant baculoviruses were used to express the gp160 envelope glycoprotein of the human immunodeficiency virus type 1 (HIV-1) and a truncated variant designated gp160(t) which lacks a transmembrane domain. Glycosylation, proteolytic cleavage, secretion, and biological activities of gp160 and gp160(t) 160(t) were studied in Spodoptera frugiperda cells. Both proteins were rapidly glycosylated and initially were found to be totally endo-beta-N-acetyl-D-glucosaminidase H (endo-H) sensitive. However, partial resistance to endo-H was gradually acquired by both molecules. gp160 was found to remain cell-associated, whereas gp160(t) was secreted into the culture medium in large amounts. A fraction of gp160 and gp160(t) appeared to be proteolytically cleaved, and a cleavage product corresponding in size to gp120 was identified in the culture medium. gp160(t) was found to interact specifically with CD4 receptors without any requirement for proteolytic cleavage. The gp160 protein was shown to be expressed on the surface of S. frugiperda cells by direct immunofluorescence. These surface molecules were biologically active, as demonstrated by their ability to induce syncytium formation when cocultivated with HeLa T4 cells.


A system for stable indirect immobilization of multimeric recombinant proteins.
To perform an ELISA or for panning phage particles which display recombinant proteins, one of the reactants is immobilized on solid phase. Immobilization in ELISA is generally performed by passive adsorption of ligands to plastic. However, protein is denatured during the adsorption process. This may result in low efficiency interaction between ligands and receptors which depend on native structures. In contrast, indirect immobilization has been shown to prevent protein denaturation. The aim was to develop a system that allows efficient and stable indirect immobilization of a variety of recombinant multimeric proteins to solid phase. A new vector was constructed which allows the expression of up to three proteins linked by the Jun/Fos leucine zipper. Purification of the resulting protein was achieved by Ni+ affinity chromatography utilizing the 6xHis-ABP (albumin binding protein) protein fused to the N-terminus of the Jun polypeptide. The high binding affinity of ABP to rat serum albumin (RSA) was exploited for indirect immobilization of recombinant proteins to solid phase. In an enzyme linked assay, the binding of ABP to immobilized RSA was shown to be 10-1000 times more efficient than other immobilization systems. Using the ZZ IgG binding domain of staphylococcal protein A as bait, the RSA-ABP immobilization system was successfully used to screen and enrich IgG Fc encoding DNA fragments from a cDNA phage library. The newly designed vector termed pJuFoexpress allows production and purification of multimeric protein complexes linked by the Jun/Fos leucine zipper. Without chemical modifications, the recombinant proteins can be immobilized indirectly to solid phase. The immobilization results in the stable display of native, biologically active proteins which can be used in ELISA and phage display systems.


Impact of secondary prevention medical therapies on outcomes of patients suffering from Myocardial Infarction with NonObstructive Coronary Artery disease (MINOCA): A meta-analysis.
To assess the impact of secondary prevention medical therapies (statins, ACE-inhibitors/Angiotensin Receptor Blockers (ARB), beta-blockers (BB) and Dual Antiplatelet Therapy (DAPT)) on outcomes of patients with myocardial infarction with nonobstructive coronary artery disease (MINOCA). Five adjusted observational studies encompassing 10,546 were included in this meta-analysis. All-cause death was the primary endpoint, while Major Adverse Cardiovascular Events (MACE) and acute myocardial infarction (AMI) were the secondary endpoints. After 24 months of follow up, statins (tested in 8093 patients) were associated with a reduced risk of all-cause death (HR 0.60:0.45-0.81, p 〈0,001), while ACE-inhibitors/ARB (on 9666 patients) were not. Aggregate data from two studies (n = 9720, 7719 on beta-blockers, 6423 on DAPT) indicated that beta-blockers and DAPT (median follow-up 34.1 and 15.7 months, respectively) were both associated with a significant reduction of all-cause death (HR0.81:0.66-0.99, p = 0.04, and HR0.73:0.55-0.98, p = 0.03, for beta-blockers and DAPT, respectively). Among the investigated therapies, only ACE-inhibitors/ARBs entailed a reduced risk of MACE (HR0.65:0.44-0.94, p = 0.02, all CI 95%) over 36.5 months (four studies, n = 10,150). None of the investigated therapies was associated with a reduced risk of AMI. Data from adjusted observational studies suggest that beta-blockers, statins and DAPT are associated with a survival benefit among MINOCA patients. ACE-inhibitors/ARB entail a reduced risk of MACE while none of the investigated secondary prevention therapies is associated with a reduced risk of AMI. Randomized controlled trials are warranted to confirm these findings.


Anti-predator behaviour of adult red-legged partridge (Alectoris rufa) tutors improves the defensive responses of farm-reared broods.
The aim of this work was to improve natural anti-predator behaviour of farm-reared gamebirds. We evaluated the anti-predator behaviour of reared red-legged partridge Alectoris rufa chicks kept in brooder houses in large groups (>350 chicks), trained and not trained by parent red-legged partridges acting as experienced tutors. The experiment consisted of two conditioned tests (a raptor model and a human) and two control tests, which were conducted during three consecutive phases of life (1-4, 15-17 and 30-32 d after hatching). The motor anti-predator behaviour, its duration, the intensity of response in chicks and alarm calls elicited by adults were recorded. Tutors elicited aerial alarm calls (76% of tests) and showed prolonged crouching (59% of tests) in response to the raptor model whereas uttering the ground alarm call (73% of tests) and showing vigilance behaviour (78% of tests) was the main pattern during the human test. Trained and not trained chicks showed similar motor behaviour in response to the raptor model (crouching) and the human test (escaping), but frequency of strong responses (all chicks responding) from chicks trained with tutors was double that of chicks trained without them, and chicks trained with tutors showed a higher frequency of long responses (41-60 s). This study indicates that anti-predator training programmes before release may improve behaviour of farm-reared partridges which may confer benefits to survival of birds.


Circulating matrix metalloproteinase-9 is transiently elevated after colorectal surgery.
Plasma levels of matrix metalloproteinases (MMPs) may yield important information in patients suffering from colorectal cancer but the effect of surgery, a common treatment modality in these patients, on circulating MMP levels is currently unknown. The aim of this study was to assess whether plasma MMP-2 and MMP-9 levels are affected by operative procedures. In total 128 patients undergoing elective surgery for colorectal cancer (n = 66), liver metastases from colorectal origin (n = 50) and arthrosis of the hip (n = 12) were included in the study. Gelatinase activity was measured, using quantitative gelatin zymography, in plasma obtained before operation and 1 week, 1 month and 3 months postoperatively. One week after operation a significant increase in proMMP-9 activity was measured after colorectal surgery (260%, p = 0.0038), liver surgery (285%, p < 0.0001) and hip surgery (217%, p = 0.012) as compared with preoperative levels. After 1 month proMMP-9 activity had returned to preoperative levels. No effect on proMMP-2 activity was measured. Operative procedures have a profound but transient effect on plasma MMP-9 activity. If used to assess disease status, postoperative plasma MMP levels should be interpreted with caution.


Hyperglycemia exacerbates colon cancer malignancy through hexosamine biosynthetic pathway.
Hyperglycemia is a common feature of diabetes mellitus, considered as a risk factor for cancer. However, its direct effects in cancer cell behavior are relatively unexplored. Herein we show that high glucose concentration induces aberrant glycosylation, increased cell proliferation, invasion and tumor progression of colon cancer. By modulating the activity of the rate-limiting enzyme, glutamine-fructose-6-phosphate amidotransferase (GFAT), we demonstrate that hexosamine biosynthetic pathway (HBP) is involved in those processes. Biopsies from patients with colon carcinoma show increased levels of GFAT and consequently aberrant glycans' expression suggesting an increase of HBP flow in human colon cancer. All together, our results open the possibility that HBP links hyperglycemia, aberrant glycosylation and tumor malignancy, and suggest this pathway as a potential therapeutic target for colorectal cancer.


Early Antibiotic Exposure Is Not Detrimental to Therapeutic Effect from Immunotherapy in Hepatocellular Carcinoma.
Immune checkpoint inhibitor (ICI) therapy is an expanding therapeutic option for hepatocellular carcinoma (HCC). Antibiotics (ATB) taken prior to or early during ICI therapy can impact immunotherapy efficacy across indications; however, the effect of ATB is undefined in HCC. In a large international cohort of 450 ICI recipients from Europe, North America, and Asia, we categorized patients according to timing of ATB focusing on exposure within -30 to +30 days from ICI (early immunotherapy period [EIOP]). EIOP was evaluated in association with overall survival (OS), progression-free survival (PFS), and best radiologic response using RECIST 1.1 criteria. Our study comprised mostly cirrhotic (329, 73.3%) males (355, 79.1%) with a Child-Turcotte Pugh class of A (332, 73.9%), receiving ICI after 1 therapy line (251, 55.9%) for HCC of Barcelona clinic liver cancer stage C (325, 72.4%). EIOP (n = 170, 37.9%) was independent of baseline clinicopathologic features of HCC and correlated with longer PFS (6.1 vs. 3.7 months, log-rank p = 0.0135). EIOP+ patients had similar OS, overall response, and disease control rates (DCRs) compared to EIOP. The effect of EIOP persisted in landmark time analyses and in multivariable models, confirming the independent predictive role of EIOP in influencing PFS following adjustment for covariates reflective of tumor burden, liver function, and ICI regimen administered. In patients receiving programmed cell death-1 receptor/ligand inhibitors monotherapy, EIOP was also associated with higher DCRs (61.4% vs. 50.9%, p = 0.0494). Unlike other oncological indications, ATB in the 30 days before or after ICI initiation is associated with improved benefit from immunotherapy, independent of disease and treatment-related features. Evaluation of the immune microbiologic determinants of response to ICI in HCC warrants further investigation.


Structure-based discovery of a new class of Hsp90 inhibitors.
Docking-based virtual screening identified 1-(2-phenol)-2-naphthol compounds as a new class of Hsp90 inhibitors of low to sub-micromolar potency. Here we report the binding affinities and cellular activities of several members of this class. A high resolution crystal structure of the most potent compound reveals its binding mode in the ATP binding site of Hsp90, providing a rationale for the observed activity of the series and suggesting strategies for developing compounds with improved properties.


Gas chromatographic-mass spectrometric study of deacetylation and oxidation of 2-acetylaminofluorene by rat liver epithelial cell lines upon cocarcinogen induction.
Cell line cultures from postnatal and adult rats were incubated with 5-100 mumol/l [9-14C]-2-acetylaminofluorene. On incubation of 10 mumol/l, ring-hydroxylated metabolites, expressed as nmol hydroxy-2-acetylaminofluorene (OH-2-AAF)/mg cell protein/24 h, were 9-OH- 1.28 +/- 0.37, 7-OH- 1.08 +/- 0.28 and 5-OH- 0.30 +/- 0.08, and deacetylated 2-AAF (2-AF) 1.20 +/- 0.18. For 5, 10, 50 and 100 mumol/l 2-AAF, the total production of OH-2-AAF (same units) and 2-AF (%) were, respectively, 0.86 (0%), 3.86 (35%), 17.8 (60%) and 35.03 (89%). On preincubation with phenobarbital (BP) or 3-methylcholanthrene (3-MC) and then incubation of 10 mumol/l 2-AAF, the total synthesis of OH-2-AAF increased 1.9-fold (PB) and 2.5-fold (3-MC). In addition, four other OH-2-AAF (1-OH-, 3-OH- and two unknown OH-2-AAF) were produced and glucuronidation of all metabolites was induced and amounted to 57% of the total after PB and 75% after 3-MC preincubation. Metyrapone or alpha-naphthoflavone inhibition of BP or 3-MC, respectively, markedly affected the production of free and conjugated metabolites and, almost completely, the deacetylation of 2-AAF.


Everolimus reduces postoperative arthrofibrosis in rabbits by inducing autophagy-mediated fibroblast apoptosis by PI3K/Akt/mTOR signaling pathway.
To investigate the effects of everolimus (EVE) on postoperative fibrosis in the knee joint and the potentially relevant signaling pathways. CCK-8 and flow cytometry assays were used to detect the effect of EVE on human fibroblast viability and apoptosis induction. IF and TEM were used to assess fibroblast autophagy. 3-methyladenine (3-MA) was applied to inhibit autophagy to clarify the relationship between autophagy and apoptosis. WB was used to measure the expression of proteins related to apoptosis, autophagy and the mTOR signaling pathway. A rabbit model of knee joint fibrosis was established and topically treated with various concentrations of EVE. IF-P was applied to identify that the main components cells of the fibrotic tissue and histomorphological staining was used to detect the degree of fibrosis and the content of collagen. Histomorphological staining demonstrated that EVE could reduce the degree of postoperative fibrosis and collagen deposition in the knee joint. The results of IF, TEM, flow cytometry assays and WB detection showed that EVE could activate autophagy and induce fibroblasts apoptosis. Meanwhile, the expression levels of p-PI3K, p-Akt, p-mTOR were downregulated with EVE treatment. After the inhibition of autophagy by 3-MA treatment, the increased fibroblasts apoptosis by EVE treatment was partially decreased. Everolimus can reduce surgery-induced knee fibrosis by inducing autophagy-mediated fibroblast apoptosis, which may be involved with the regulation of the PI3K/Akt/mTOR signaling pathway.


Insights into Anion-Solvent Interactions to Boost Stable Operation of Ether-Based Electrolytes in Pure-SiOx ||LiNi0.8 Mn0.1 Co0.1 O2 Full Cells.
Ether solvents with superior reductive stability promise excellent interphasial stability with high-capacity anodes while the limited oxidative resistance hinders their high-voltage operation. Extending the intrinsic electrochemical stability of ether-based electrolytes to construct stable-cycling high-energy-density lithium-ion batteries is challenging but rewarding. Herein, the anion-solvent interactions were concerned as the key point to optimize the anodic stability of the ether-based electrolytes and an optimized interphase was realized on both pure-SiOx anodes and LiNi0.8 Mn0.1 Co0.1 O2 cathodes. Specifically, the small-anion-size LiNO3 and tetrahydrofuran with high dipole moment to dielectric constant ratio realized strengthened anion-solvent interactions, which enhance the oxidative stability of the electrolyte. The designed ether-based electrolyte enabled a stable cycling performance over 500 cycles in pure-SiOx ||LiNi0.8 Mn0.1 Co0.1 O2 full cell, demonstrating its superior practical prospects. This work provides new insight into the design of new electrolytes for emerging high-energy density lithium-ion batteries through the regulation of interactions between species in electrolytes.


The metabolism of menadione (2-methyl-1,4-naphthoquinone) by isolated hepatocytes. A study of the implications of oxidative stress in intact cells.
The cytotoxic effects of many quinones are thought to be mediated through their one-electron reduction to semiquinone radicals, which subsequently enter redox cycles with molecular oxygen to produce active oxygen species and oxidative stress. The two-electron reduction of quinones to diols, mediated by DT-diaphorase (NAD(P)H: (quinone-acceptor) oxidoreductase), may therefore represent a detoxifying pathway which protects the cell from the formation of these reactive intermediates. By using menadione (2-methyl-1,4-naphthoquinone) and isolated hepatocytes, the relative contribution of the two pathways to quinone metabolism has been studied and a protective role for DT-diaphorase demonstrated. Moreover, in the presence of cytotoxic concentrations of menadione rapid changes in intracellular thiol and Ca2+ homeostasis were observed. These were associated with alterations in the surface structure of the hepatocytes which may be an early indication of cytotoxicity.


Clinical research in surgery: questions but few answers.
Surgeons must be prepared to be questioned about their understanding, activities, and achievements in basic and clinical research in terms of structural, organizational, and financial support of surgical research. Clinical research today comprises basic research related to accumulation of knowledge about biological systems, disease-related research including experimental research with animals, and in vitro research with biological material and clinical studies with the strongest empirical basis in data derived from controlled clinical trials. Most clinical methods related to diagnosis and treatment have been introduced into the practice of everyday medicine even in university hospitals without passing a prospective evaluation protocol. To test the efficacy of a new drug or a newly developed surgical technique, the controlled prospective clinical trial is the best method; however, fewer than 50% of clinical questions can be answered by controlled clinical studies. In Germany the frequency of controlled clinical trials for answering clinical questions is presently low; this is particularly true for surgery, in comparison with other countries. The concept of evidence-based medicine is based on the principles of data generation from controlled trials and meta-analyses of those studies. Best-evidence synthesis means a reasonable synthesis of evident knowledge, experience and intuition. The challenge of professionalization of clinical research in surgery is linked to the establishment of full time positions for basic scientists and clinical researchers and the establishment of clinical research teams and of research professorships including basic scientists in surgical departments. Clinical research can be effective only if there is good cooperation between clinicians and basic scientists, if high grade scientific methods are established in clinical research institutions, if there is sufficient financial and personal supply, and if there are enough laboratories and animal operating facilities. There is an urgent need for structures integrating basic scientists in clinical research groups, for more interdisciplinary research projects and more multidisciplinary research, independent of clinical care in terms of manpower.


Kinect-based objective assessment for early frailty identification in patients with Parkinson's disease.
Frailty is common in Parkinson's disease (PD) and increases vulnerability to adverse outcomes. Early detection of this syndrome aids in early intervention. To objectively identify frailty at an early stage during routine motor tasks in PD patients using a Kinect-based system. PD patients were recruited and assessed with the Fried criteria to determine their frailty status. Each participant was recorded performing the Movement Disorder Society-Sponsored Revision of the Unified Parkinson's Disease Rating Scale part III (MDS-UPDRS III) extremity tasks with a Kinect-based system. Statistically significant kinematic parameters were selected to discriminate the pre-frail from the non-frail group. Of the fifty-two participants, twenty were non-frail and thirty-two were pre-frail. Decreased frequency in finger tapping (P = 0.005), hand grasping (P = 0.002), toe tapping (P = 0.002), and leg agility (P = 0.019) alongside reduced hand grasping speed (P = 0.030), lifting (P < 0.001) and falling speed (P < 0.001) in leg agility were observed in the pre-frail group. Amplitude in leg agility (P = 0.048) and amplitude decrement rate (P = 0.046) in hand grasping showed marginally significant differences between two groups. Moderate discriminative values were found in frequency and speed of the extremity tasks to identify pre-frailty with sensitivity, specificity, and area under the curve (AUC) in the range of 45.00-85.00%, 68.75-100%, and 0.701-0.836, respectively. The combination of frequency and speed in extremity tasks showed moderate to high discriminatory ability, with AUC of 0.775 (95% CI 0.637-0.913, P < 0.001) for upper limb tasks and 0.909 (95% CI 0.832-0.987, P < 0.001) for lower limb tasks. When combining these features in both upper and lower limb tasks, the AUC increased to 0.942 (95% CI 0.886-0.999, P < 0.001). Our findings demonstrated the promise of utilizing Kinect-based kinematic data from MDS-UPDRS III tasks as early indicators of frailty in PD patients.


Whole genome sequencing data of Oryza sativa L. cv. Takanari.
Oryza sativa L. 'Takanari' is one of the most productive indica cultivars [1,2]. Reciprocal chromosome segment substitution lines (CSSLs) derived from a cross between 'Koshihikari' and 'Takanari' are useful tools for the detection and precise mapping of target quantitative trait loci (QTL) in 'Takanari'. Although the available Os-Nipponbare-Reference-IRGSP-1.0 reference genome is available and useful for evaluating genetic diversity among japonica cultivars, it is not always useful for evaluating genetic diversity harbored by indica cultivars such as 'Takanari'. To reveal sequence variants in 'Takanari' and to exploit these variants in rice breeding programs, the whole genome of 'Takanari' was sequenced using a combination of Illumina HiSeq X Ten (20,983,495 reads and %GC 43) and PacBio (2,847,220 high-quality subreads). NGS data obtained have been deposited in the DNA Data Bank of Japan (DDBJ) under accession number DRA007557.


Perceived psychosocial impairment associated with eating disorder features: responses to a mental health literacy intervention.
Whether and to what extent young adults are aware of the adverse impact of eating disorder features (EDF) on psychosocial functioning is unclear, although such awareness may affect the experience and behavior of sufferers. The aim of the current study was to examine young adults' perceptions of psychosocial impairment associated with EDF, and the potential effect on these perceptions of an eating disorders "mental health literacy" (ED-MHL) intervention. Undergraduate students (male: n = 35; female: n = 141) completed self-report questionnaires prior to, immediately following, and 3 months after completion of a 3-h ED-MHL intervention. Perceived psychosocial impairment associated with EDF-binge eating, purging, extreme dietary restriction, overvaluation of weight/shape, and excessive exercise-was assessed at each time point. At all 3 time points, EDF were considered to have a 'slightly negative' to 'very negative' impact on psychosocial functioning. Prior to the intervention, binge eating, purging and extreme dietary restriction were generally considered to have a greater negative impact than excessive exercise and overvaluation of weight/shape. Three months after the ED-MHL intervention, participants reported greater perceived impairment associated with excessive exercise and overvaluation; while perceptions of psychosocial impairment associated with binge eating, purging and dietary restriction remained largely unchanged. Females perceived greater impairment associated with EDF than males did immediately after the intervention, but not at the 3-month follow-up. The adverse effects on psychosocial functioning of binge eating, purging and extreme dietary restriction appear to be readily recognized by young people. Awareness of the adverse effects of excessive exercise and overvaluation may be poorer, but amenable to improvement by means of a relatively simple intervention. These features may warrant particular attention in health promotion programs.


Enhancing the Cellular Production of Extracellular Vesicles for Developing Therapeutic Applications.
Extracellular vesicles (EVs) have various advantageous properties, including a small size, high biocompatibility, efficient cargo loading, and precise cell targeting ability, making them promising tools for therapeutic development. EVs have been increasingly explored for applications like drug delivery. However, due to limited cellular secretion rates of EVs, wide-scale clinical applications are not achievable. Therefore, substantial strategies and research efforts have been devoted to increasing cellular secretion rates of EVs. This review describes various studies exploring different methods to increase the cellular production of EVs, including the application of electrical stimulus, pharmacologic agents, electromagnetic waves, sound waves, shear stress, cell starvation, alcohol, pH, heat, and genetic manipulation. These methods have shown success in increasing EV production, but careful consideration must be given as many of these strategies may alter EV properties and functionalities, and the exact mechanisms causing the increase in cellular production of EVs is generally unknown. Additionally, the methods' effectiveness in increasing EV secretion may diverge with different cell lines and conditions. Further advancements to enhance EV biogenesis secretion for therapeutic development is still a significant need in the field.


Hybridization of short-range and long-range charge transfer excited states in multiple resonance emitter.
Multiple resonance (MR) thermally activated delayed fluorescence emitters have been actively studied as pure blue dopants for organic light-emitting diodes (OLEDs) because of excellent color purity and high efficiency. However, the reported MR emitter, 2,5,13,16-tetra-tert-butylindolo[3,2,1-jk]indolo[1',2',3':1,7]indolo[2,3-b]carbazole (tDIDCz) based on bis-fused indolocarbazole framework could not demonstrate efficient triplet-to-singlet spin crossover. In this work, we report two isomeric MR emitters designed to promote triplet exciton harvesting by reconstructing the electronic structure of tDIDCz. To manage excited states, strong electron donors were introduced at the 2,5-/1,6-position of tDIDCz. As a result, 2,5-positions managed tDIDCz shows long-range charge transfer characteristics while preserving the MR nature. Quantum chemical calculation demonstrates direct spin-orbit coupling by long-range charge transfer and spin-vibronic coupling assisted reverse intersystem crossing by short-range charge transfer simultaneously contribute to triplet-to-singlet spin crossover. Consequently, high performance blue OLED recorded a high external quantum efficiency of 30.8% at a color coordinate of (0.13, 0.13).


Safe and acceptable strategies for producing foreign molecules in plants.
The ability to express foreign genes using transgenic technologies has opened up options for producing large quantities of commercially important industrial or pharmaceutical products in plants. These technologies have made it possible to use well-developed systems of commercial agriculture that were developed principally to produce raw material for large-scale food, feed or processing applications for the production of foreign molecules. The possibility of the novel industrial or pharmaceutical molecules produced in such plants, or components derived from them, contaminating the environment and food chains has become especially controversial. This potential contamination has prompted detailed consideration of how such crops and the molecules that they produce can be effectively isolated and contained. First, the crop can be completely isolated physically from its food or feed counterpart during every aspect of its development and commercialization. Second, genetic isolation systems or genetic barriers that prevent normal reproduction can be used to reduce the likelihood of the industrial or pharmaceutical crop entering the food chain.


Exploring the use of social network analysis to measure communication between disease programme and district managers at sub-national level in South Africa.
With increasing interest in maximising synergies between disease control programmes (DCP) and general health services (GHS), methods are needed to measure interactions between DCP and GHS actors. In South Africa, administrative integration reforms make GHS managers at decentralised level (district managers) responsible for the oversight of DCP operations within districts, with DCP managers (programme managers) providing specialist support. The reforms necessitate interdependence, but these actors work together ineffectively. Communication is crucial for joint working, but no research to assess communication between these actors has been done. This study explores the use of social network analysis (SNA) to measure the extent to which programme and district managers in South Africa communicate, using HIV monitoring and evaluation (M&E) as an exemplar. Data were collected from fifty one managers in two provinces during 2010-2011, to measure: a) one-on-one task-related communication - talking about the collation (verification, reporting) and use of HIV data for monitoring HIV interventions; and b) group communication through co-participating in management committees where HIV data are used for monitoring HIV interventions in districts. SNA measures were computed to describe actor centrality, network density (cohesion), and communication within and between respective manager groups. Block modelling was applied to identify management committees that connect respective manager groups. Results show HIV programme managers located at higher level communicated largely amongst themselves as a group (homophily), seldom talked to the district managers to whom they are supposed to provide specialist HIV M&E support, and rarely participated with them in management committees. This research demonstrates the utility of SNA as a tool for measuring the extent of communication between DCP and GHS actors at sub-national level. Actions are needed to bridge observed communication gaps in order to promote collaborative monitoring of HIV programme interventions within districts.


A study of Epstein-Barr virus infection in the Chinese tree shrew(Tupaia belangeri chinensis).
Epstein-Barr virus (EBV) is closely associated with many human diseases, including a variety of deadly human malignant tumours. However, due to the lack of ideal animal models,the biological characteristics of EBV, particularly its function in tumourigenesis, have not been determined. Chinese tree shrews (Tupaia belangeri chinensis), which are similar to primates, have been used to establish a variety of animal models and have recently received much attention. Here, we established tree shrews as a model for EBV infection by intravenous injection. Ten tree shrews were inoculated with EBV by intravenous injection,and blood was collected at regular intervals thereafter from the femoral artery or vein to detect EBV markers. Eight of 10 tree shrews showed evidence of EBV infection. In the 8 EBV-infected tree shrews, EBV copy number increased intermittently or transiently, EBV-related gene expression was detected, and anti-EBV antibodies increased to varying degrees. Macroscopic hepatomegaly was observed in 1 tree shrew, splenomegaly was observed in 4 tree shrews, and enlarged mesenteric lymph nodes were observed in 3 tree shrews. Haematoxylin and eosin (HE) staining showed splenic corpuscle hyperplasia in the spleens of 4 tree shrews and inflammatory cell infiltration of the liver of 1 tree shrew and of the mesenteric lymph nodes of 3 tree shrews. EBER in situ hybridization(ISH) and immunohistochemical (IHC) staining showed that EBER-, LMP1- and EBNA2- positive cells were present in the spleens and mesenteric lymph nodes of some tree shrews. Western blotting (WB) revealed EBNA1-positive cells in the spleens of 4 tree shrews. EBV markers were not detected by HE, EBER-ISH or IHC in the lung or nasopharynx. These findings suggest that EBV can infect tree shrews via intravenous injection. The presented model offers some advantages for exploring the pathophysiology of EBV infection in humans.


Multiplex proteomics as risk predictor of infection in patients treated with hemodialysis-A prospective multicenter study.
Severe infection is a major problem in hemodialysis patients. Multiplex proteomics might reveal novel insights into disease mechanisms increasing the risk of infection and might also be used as a risk prediction tool. The aims of this study were (1) to evaluate associations between 92 proteins assessed by a proximity extension assay and the development of severe infection in patients on hemodialysis and (2) to develop a risk prediction model for severe infection using prespecified clinical variables and proteomics. Prospective, observational multicenter cohort study with 5-year follow-up. Patients receiving in-center hemodialysis in five facilities in Denmark were included. The primary composite endpoint was death caused by infection, bacteremia, and infections requiring hospitalization of at least 2 days or prolonging a hospital stay. Of 331 patients included 210 patients reached the primary endpoint during follow-up. In adjusted Cox regression analyses, 14 plasma proteins were associated with severe infection. Correcting for multiple testing revealed only cathepsin-L1 and interleukin-6 significantly associated with the primary outcome. Cathepsin-L1-hazard ratio: 1.64 (95% confidence interval [CI] 1.24-2.17) and interleukin-6-hazard ratio: 1.16 (95% CI 1.05-1.29). Apparent C-statistics of the risk prediction model using clinical variables was 0.605, addition of cathepsin-L1 and interleukin-6 to the model improved discrimination slightly: C = 0.625. Proteomic profiling identified cathepsin-L1 and interleukin-6 as markers for infectious risk in hemodialysis patients. Further studies are needed to replicate the results and to examine possible causality. The developed risk prediction models need considerable improvement before implementation in clinical practice is meaningful.


[Supporting the clinical assessment: from a printed handbook to its digital version].
Nurses from a palliative care network questioned the methods of the global clinical assessment performed in patients' homes. Going back to the very foundations of the profession, they drew up a handbook on this subject aimed at home care professionals. An embedded digital version has been developed enabling caregivers to consult it at any time on their mobile telephone.


First 102 patients with the BioValsalva conduit for aortic root replacement.
We retrospectively evaluated our results with the BioValsalva Conduit (Vascutek Terumo, Renfrewshire Scotland), a stentless porcine valve incorporated in a 3-layered prosthetic graft. From July 2008 through April 2011, 102 patients with a mean age of 70.9 ± 7.3 years underwent aortic root replacement with a BioValsalva conduit. The indication for surgery was aneurysmal disease of the aorta in 81 patients (79.4%), aortic valve endocarditis in 15 patients (14.7%), acute type A aortic dissection in 4 patients (3.9%), and other causes in 2 patients (2.0%). In 26 patients (25.5%), the intervention was a reoperation. Overall hospital mortality was 4.9% (n = 5; 95% confidence limit [CL], 1.6%-11.1%). Cause of death was cardiac failure in 2 patients, multiple organ or renal failure in 2 patients, and tamponade in 1 patient. Mean follow-up was 8.1 months. During follow-up, 3 deaths occurred (3.1%) because of mediastinitis, cardiac ischemia, and arrhythmia. The overall survival at 3 and 12 months was 95.9% (95% CL, 92.0% -99.9%) and 92.1% (95% CL, 85.7% -98.9%) respectively. Three patients (3.1%) had new-onset endocarditis of the BioValsalva conduit; 2 of these patients required reoperation and 1 patient received antibiotic treatment only. Retrospective analysis of the BioValsalva conduit for aortic root replacement in more than 100 consecutive patients demonstrated satisfactory initial results, with low mortality and acceptable low morbidity rates. Follow-up is mandatory and long-term results are to be awaited.


Nasopharyngeal Stenosis: A Rare Radiotherapy Complication.
Acquired nasopharyngeal stenosis after radiotherapy for NPC is rare. We report a case of bilateral choanal stenosis and nasopharygeal stenosis in patient treated with chemo radiotherapy for undifferentieted nasopharnygeal carcinoma. A 68 years old man treated for undifferentiated nasopharyngeal carcinoma with chemo-radiotherapy 10 years before. He complained of complete nasal obstruction, rhinorrhea, anosmia, snoring and hearing loss. Clinical examination and imaging revealed bilateral choanal stenosis and nasopharyngeal stenosis. Patient underwent a transnasal endoscopic resection for scar tissue with stenting of posterior choanae. Restenosis was noted just after stent removal. Second endoscopic resection of stenosis was done, followed by application of mitomycin-C without stenting. Several months later, the patient has good airflow, a patent nasopharynx without reccurence of stenosis. The use of topical mitomycin-C may prove useful in the treatment and prevention of subsequent restenosis and scar formation in larynx and tracheal. Mitomycin-C application has being shown useful in treating nasopharyngeal stenosis. It is safely used topically in various surgical sites without major side effect. Nasopharyngeal stenosis is an unusual complication of radiotherapy that can be successfully treated with combine two modalities: surgical and mitomycin-C as an adjunct.


Effect of pulmonary hypertension on lung compliance in newborn lambs.
We studied the effects of increased pulmonary arterial pressure and vascular resistance on lung compliance in 12 anesthetized newborn lambs. Lung compliance was unaffected by large changes in pulmonary artery pressure and pulmonary vascular resistance (80% increase) that were induced by either hypoxia or acidosis. We conclude that contraction of the pulmonary vascular smooth muscle in normal newborn lambs does not affect the stiffness of the lung in the parenchymal regions undergoing volume change during ventilation.


The prevalence of depressive symptoms among the older in China: a meta-analysis.
This study aims to find the status and prevalence of depressive symptoms among the population aged 60 years and above in China. We conducted a meta-analysis of cross-sectional studies in older population. Epidemiological studies on depressive symptoms published in Chinese journals were identified manually and online by using the Chinese Biological Medical Literature Database, Chongqing VIP database for Chinese Technical Periodicals, and Chinese National Knowledge Infrastructure database. Studies published in English journals were identified using Medlars Online. A total of 32 eligible studies were included in this review. The analysis showed that the pooled prevalence (with 95% confidence interval) of depressive symptoms for the seniors in China was 22.7% (confidence interval: 19.4-26.4%). The prevalence of depressive symptoms among the older was a little higher in women (24.2%) than in men (19.4%), in rural (29.2%) than urban (20.5%) areas, and in western (30.5%) than eastern (19.5%) areas. In addition, the prevalence of depressive symptoms decreased with increasing levels of education. However, it did not increase gradually with age. The prevalence of depressive symptoms among the older in China was a little high, and it was varied by gender, age, area, and education level.


The use of monoclonal antibodies for the characterization of a 5 S gene-specific transcription factor (IIIA) from Xenopus laevis.
Monoclonal antibodies were raised against a 5 S gene-specific transcription factor (TFIIIA) from Xenopus laevis oocytes. Seven stable hybridoma lines were established and shown to secrete antibodies directed against different antigenic structures present on distinct peptides of the factor (generated by cleavage of TFIIIA with cyanogen bromide). All antibodies react with denatured TFIIIA in nitrocellulose blots, but only five are able to bind to the native protein. These antibodies recognize the transcription factor not only in its purified (RNA-free) form, but also when it is complexed with 5 S RNA in 7 S ribonucleoprotein complexes. None of the monoclonal antibodies inhibits transcription of 5 S genes in vitro, although an interaction of the antibodies with the factor in a transcription reaction could be demonstrated by a depletion assay. Thus, binding of the antibodies to TFIIIA does not interfere with the function of the factor during transcription initiation. Additionally, it was found that transcriptionally active TFIIIA could be purified to greater than 95% homogeneity from ovarian extracts using an antibody coupled to Sepharose.


Retracted: Innovative Practice of Music Education in the Universities in the Context of 5G Network.
[This retracts the article DOI: 10.1155/2022/3451422.].


Comparative Analysis of Aggregation of Thermus thermophilus Ribosomal Protein bS1 and Its Stable Fragment.
Functionally important multidomain bacterial protein bS1 is the largest ribosomal protein of subunit 30S. It interacts with both mRNA and proteins and is prone to aggregation, although this process has not been studied in detail. Here, we obtained bacterial strains overproducing ribosomal bS1 protein from Thermus thermophilus and its stable fragment bS1(49) and purified these proteins. Using fluorescence spectroscopy, dynamic light scattering, and high-performance liquid chromatography combined with mass spectrometric analysis of products of protein limited proteolysis, we demonstrated that disordered regions at the N- and C-termini of bS1 can play a key role in the aggregation of this protein. The truncated fragment bS1(49) was less prone to aggregation compared to the full-size bS1. The revealed properties of the studied proteins can be used to obtain protein crystals for elucidating the structure of the bS1 stable fragment.


Artemiprinolides A-M, thirteen undescribed sesquiterpenoid dimers from Artemisia princeps and their antihepatoma activity.
Bioassay-guided investigation of the active fraction of Artemisia princeps led to 13 undescribed sesquiterpenoid dimers, artemiprinolides A-M (1-13), together with 11 known ones (14-24). Their structures were elucidated by comprehensive spectroscopic data and absolute configurations were assigned based on single crystal X-ray diffraction data and ECD calculations. Structurally, all compounds were postulated to be derived from the Diels-Alder cycloaddition. The isolated dimers except 11 and 15 were assayed for their cytotoxicity against HepG2, Huh7, and SK-Hep-1 cell lines, of which four compounds (3, 13, 17, 18) exhibited obvious cytotoxicity with IC50 values ranging from 8.8 to 20.1 μM. Interestingly, the most active compounds 1 and 16 manifested significant cytotoxicity on the three tested hepatoma cell lines with IC50 values of 5.4, 4.1 (HepG2), 7.7, 5.6 (Huh7), and 11.8, 15.7 μM (SK-Hep-1), respectively, which were better than sorafenib. Compound 1 dose-dependently inhibited cell migration and invasion, and significantly induced the HepG2 cell arrest in G2/M phase by downregulating cdc2 and pcdc2 and upregulating cyclinB1; and induced apoptosis by downregulating Bcl-2 expression and upregulating Bax level. The molecular docking study implied that the carbonyl at the C-12' of 1 had a strong binding affinity with PRKACA.


[Anatomy of the superior labrum complex of the shoulder].
The outstanding progress achieved in MRI and arthroscopic techniques has forced orthopedic surgeons to consider the anatomy of the shoulder joint in detail. The superior labrum complex shows a broad range of variations, which is the reason for some difficulties regarding diagnostic conclusions as well as therapeutic considerations. In particular, the differentiation between the normal and pathological adherence of the labrum is a serious clinical problem. The glenoid labrum consists of circularly arranged collagenous fibers attached to the osseous margin of the glenoid by a fibrocartilaginous transitional zone. The cranial part of the labrum is more compact than the caudal one. It is also connected to the joint capsule and the integrated ligaments. Most of the fibers of the long head of the biceps tendon insert at the supraglenoid tubercle, only some of them continue in the fibrous ring. So-called SLAP lesions have to be differentiated from variations. In up to 18% of the patients normal clefts can be seen, e. g., sublabral holes, and in less than 6% other rare variations, e. g., the so-called Buford complex.


The glucocorticoid triamcinolone acetonide inhibits osmotic swelling of retinal glial cells via stimulation of endogenous adenosine signaling.
The glucocorticoid triamcinolone acetonide is clinically used for the treatment of macular edema. However, the edema-resolving mechanisms of triamcinolone are incompletely understood. Since cell swelling is a central cause of cytotoxic edema in the brain and retina, we determined the effects of triamcinolone acetonide on the swelling of retinal ganglion and Müller glial cells in acutely isolated retinas from rats and guinea pigs in situ. Triamcinolone acetonide (100 microM) had no effect on the swelling of ganglion cells that was evoked in isolated whole mounts of the guinea pig retina by acute application of glutamate (1 mM) or high K+ (50 mM). However, triamcinolone reversed the osmotic swelling of Müller glial cells in retinas of the rat that was observed under various experimental conditions: in retinas isolated at 3 days after transient retinal ischemia, in retinas of eyes with lipopolysaccharide-induced ocular inflammation, and in control retinas in the presence of Ba2+ (1 mM), H2O2 (200 microM), arachidonic acid (10 microM), or prostaglandin E2 (30 nM). The inhibiting effect of triamcinolone on osmotic glial cell swelling was mediated by stimulation of transporter-mediated release of endogenous adenosine and subsequent A1 receptor activation, resulting in an elevation of the intracellular cAMP level and activation of the protein kinase A, and, finally, in an opening of extrusion pathways for K+ and Cl- ions. The inhibitory effect on the cytotoxic swelling of glial cells may contribute to the fast edema-resolving effect of vitreal triamcinolone observed in human patients.


VX-680, a potent and selective small-molecule inhibitor of the Aurora kinases, suppresses tumor growth in vivo.
The Aurora kinases are essential for the regulation of chromosome segregation and cytokinesis during mitosis. Aberrant expression and activity of these kinases occur in a wide range of human tumors, and lead to aneuploidy and tumorigenesis. Here we report the discovery of a highly potent and selective small-molecule inhibitor of Aurora kinases, VX-680, that blocks cell-cycle progression and induces apoptosis in a diverse range of human tumor types. This compound causes profound inhibition of tumor growth in a variety of in vivo xenograft models, leading to regression of leukemia, colon and pancreatic tumors at well-tolerated doses. Our data indicate that Aurora kinase inhibition provides a new approach for the treatment of multiple human malignancies.


Prediction of binding modes and affinities of 4-substituted-2,3,5,6-tetrafluorobenzenesulfonamide inhibitors to the carbonic anhydrase receptor by docking and ONIOM calculations.
Inhibition activities of a series of 4-substituted-2,3,5,6-tetrafluorobenzenesulfonamides against the human carbonic anhydrase II (HCAII) enzyme have been explored by employing molecular docking and hybrid QM/MM methods. The docking protocol has been employed to assess the best pose of each ligand in the active site cavity of the enzyme, and probe the interactions with the amino acid residues. The docking calculations reveal that the inhibitor binds to the catalytic Zn(2+) site through the deprotonated sulfonamide nitrogen atom by making several hydrophobic and hydrogen bond interactions with the side chain residues depending on the substituted moiety. A cross-docking approach has been adopted prior to the hybrid QM/MM calculation to validate the docked poses. A correlation between the experimental dissociation constants and the docked free energies for the enzyme-inhibitor complexes has been established. Two-layered ONIOM calculations based on QM/MM approach have been performed to evaluate the binding efficacy of the inhibitors. The inhibitor potency has been predicted from the computed binding energies after taking into account of the electronic phenomena associated with enzyme-inhibitor interactions. Both the hybrid (B3LYP) and meta-hybrid (M06-2X) functionals are used for the description of the QM region. To improve the correlation between the experimental biological activity and the theoretical results, a three-layered ONIOM calculation has been carried out and verified for some of the selected inhibitors. The charge transfer stabilization energies are calculated via natural bond orbital analysis to recognize the donor-acceptor interaction in the binding pocket of the enzyme. The nature of binding between the inhibitors and HCAII active site is further analyzed from the electron density distribution maps.


Medicaid Expansion and Mechanical Ventilation in Asthma, Chronic Obstructive Pulmonary Disease, and Heart Failure.
Rationale: The Affordable Care Act's Medicaid expansion has led to increased access to chronic disease care among newly insured adults. Despite this, its effects on clinical outcomes, particularly for patients with asthma, chronic obstructive pulmonary disease, and heart failure, are uncertain. Objectives: To assess whether Medicaid expansion was associated with changes in mechanical ventilation rates among hospitalized patients with heart failure, asthma, and chronic obstructive pulmonary disease. Methods: Difference-in-differences analysis comparing discharge data from four states that expanded Medicaid in 2014 (Arizona, Iowa, New Jersey, and Washington) and three comparison states that did not (North Carolina, Nebraska, and Wisconsin) was performed. Models were adjusted for patient and hospital factors. Results: Mechanical ventilation rates at baseline were 7.2% in nonexpansion states and 8.8% in expansion states. Medicaid expansion was associated with a decline in mechanical ventilation rates at -0.2% per quarter (95% confidence interval [CI], -0.3% to 0.0%; P = 0.010). We did not observe a change in the rate of ICU admission (-0.4% per quarter; 95% CI, -0.8% to 0.1%; P = 0.10) or in-hospital mortality (0.1% per quarter; 95% CI, 0.0% to 0.1%; P = 0.30). In a negative control among adults aged 65 years or older, changes in mechanical ventilation rates were similar, though the CIs crossed zero (-0.1%; 95% CI, -0.2% to 0.0%; P = 0.08). Conclusions: Medicaid expansion may have been associated with a decline in mechanical ventilation rates among uninsured and Medicaid-covered patients admitted with heart failure, chronic obstructive pulmonary disease, and asthma.


Persistence of increasing vegetation gross primary production under the interactions of climate change and land use changes in Northwest China.
Substantial evidence suggests a widespread increase in global vegetation gross primary production (GPP) since the 1980s. If the increasing trend of GPP remains unchanged in the future, it is considered to be the persistence of increasing GPP. However, it is still unknown whether the vegetation increasing GPP is persistent under the interactive effects of climate change and land use changes in Northwest China. Using the Mann-Kendall and boosted regression tree models, we constructed the relationship between the increasing GPP and environmental variables, and further explored its persistence under the interactions between climate change and land use changes under SSP245 and SSP585 scenarios. The results indicated that: (1) Land use change (8.01%) was the most important variable for the increasing GPP. The surface net solar radiation (6.79%), and maximum temperature of the warmest month (6.78%) were also very important. Moreover, mean temperature of the warmest quarter had strong interactions with mean precipitation of the warmest quarter (9.82%) and land use change (8.24%). (2) Under the SSP245 scenario, the persistence of increasing GPP accounted for 65.06% of the area in 2100, mainly located in Qinghai, Ningxia, and Shaanxi, while it only accounted for 19.50% under the SSP585 scenario. (3) The SSP245 scenario moderate warming leads to a slight ecosystem benefit, with more areas developing an increase in GPP due to climate and land use change factors. On the other hand, under SSP585 scenario, there are widespread losses of increasing GPP, driven largely by climate change, while ecological engineering is conducive to the persistence of increasing GPP in southern Qinghai. The results highlight the importance of the interactive effects of climate change and land use changes for predicting the persistence of vegetation change.


Endothelium-dependent and endothelium-independent vasorelaxant effects of unripe Rubus coreanus Miq. and Dendropanax morbiferus H. Lév. extracts on rat aortic rings.
Many clinical trials on antihypertensive drugs have confirmed the usefulness of these drugs in regulating blood pressure effectively. However, all the drugs usually require long-term use; thus, economic burdens as well as some adverse effects, including headache, diarrhea, skin rash, edema, fever, and liver and kidney dysfunction, accompany their use. Therefore, we attempted to identify natural medications for treating hypertension. We investigated the antihypertensive effects of Dendropanax morbiferus H. Lév. extract (DP), enzymatically hydrolyzed DP extract (Hy-DP) and 5% unripe Rubus coreanus Miq. ethanol extract (5-uRCK). Extracts of the unripe R. coreanus were made using 20 volumes of 5% ethanol at 100 °C for 4 h. The dried leaves of D. morbiferus were subjected to enzymatic hydrolysis by protease, trypsin, bromelain and papain to increase L-arginine and GABA levels. Vasorelaxant effects of these extracts were evaluated on rat aorta precontracted with phenylephrine. In addition, hippocampal neurons, RAW 264.7 macrophages and human umbilical vein endothelial cells (HUVECs) were used to exam nitric oxide (NO) production and NO synthase (NOS) gene expression. DP, Hy-DP and 5-uRCK dose-dependently relaxed isolated rat aortic rings contracted with phenylephrine; however, Hy-DP was more effective than DP. L-NAME and ODQ differentially inhibited Hy-DP- and 5-uRCK-induced relaxation; both L-NAME and ODQ completely blocked 5-uRCK-mediated relaxation. Endothelium-denuded aortic ring relaxation was induced much less by 5-uRCK than by Hy-DP. Therefore, 5-uRCK and Hy-DP induced vascular relaxation by endothelium-dependent and partially endothelium-dependent mechanisms, respectively. Hy-DP and 5-uRCK induced eNOS gene expression and NO production in endothelial cells but did not change iNOS/nNOS expression or NO production in macrophages or neuronal cells. Both Hy-DP and 5-uRCK effectively induced vascular relaxation via similar but slightly different mechanisms. The best effective combination was investigated after mixing Hy-DP and 5-uRCK at different ratios. The 2:1 Hy-DP:5-uRCK mixture inhibited ACE, cGMP- and cAMP-dependent phosphodiesterase activity and vascular relaxation better than the other mixtures. In conclusion, Hy-DP and 5-uRCK exert antihypertensive effects through different endothelium-dependent or endothelium-independent mechanisms. These findings may greatly help elucidate the mechanisms of clinical efficacy of Hy-DP:5-uRCK mixtures used for blood pressure regulation.


Experimental and theoretical magnetostructural studies on discrete heterometallic cyanide-bridged dinuclear FeIIIMnII and tetranuclear FeIII2CuII2 complexes bearing tripodal pyrazolyl borate and tetradentate phenolate-based ligands.
Two new complexes [FeIII(Tp)(CN)2(μ-CN)MnIICl(HL1)]·3DMF (1) and {[FeIII(Tp)(CN)(μ2-NC)2CuII(HL2)](ClO4)}2·6DMF (2) (HL1 = 2-((((1-methylbenzimidazol-2-yl)methyl)(pyridin-2-ylmethyl)amino)methyl)phenol and HL2 = 2-(((pyridin-2-ylmethyl)(quinolin-2-ylmethyl)-amino)methyl)phenol) have been synthesized and characterized by elemental analysis and IR and UV/vis spectroscopy. Structural analysis revealed that 1 is a discrete dinuclear coordination complex and 2 is a discrete tetranuclear coordination complex. In complex 1, each MnII is in a distorted octahedral MN4OCl environment where coordination is satisfied by three nitrogen atoms and one oxygen atom of the ligand, and a chloride group and one nitrogen atom from cyanide. In complex 2, each Cu is in a distorted octahedral MN5O environment where coordination is satisfied by three nitrogen atoms and one oxygen atom of the ligand, and two nitrogen atoms from two cyanides. Direct current (dc) variable-temperature magnetic susceptibility measurements on polycrystalline samples of 1 and 2 were carried out in the temperature range of 1.8-300 K. Investigation of the magnetic properties reveals the occurrence of weak antiferromagnetic coupling between the low-spin FeIII (S = 1/2) ions and high-spin MnII (S = 5/2) ions in 1, while 2 exhibits ferro- and antiferromagnetic coupling between the metal ions in the tetranuclear CuII2FeIII2 unit. DFT calculations show ferromagnetic coupling in both complexes, although this appears to be weak in the case of complex 1. In addition, magnetostructural correlations reveal the magnetic behavior against Mn-N-C and Fe-C-N angles in 1 and Cu-N-C and Fe-C-N angles in 2.


Management of low back pain with facet joint injections and nerve root blocks under computed tomography guidance. A prospective study.
The aim of this work was to assess the performance of facet joint and nerve root infiltrations under computed tomography guidance for the management of low back pain and to investigate the complications and patient tolerance. The study was board-certified and informed consent was obtained from all patients. In 1 year, 86 consecutive patients (47 male, 39 female, age range 47-87 years, mean age 63) with low back pain for more than 2 years were included. All patients were clinically examined and had cross-sectional imaging performed before the procedure. Fifty-five facet joint infiltrations and 31 nerve blocks were performed under computed tomography guidance. All patients completed two valid pain questionnaires before and 3 months after the procedures. At the same time, they were clinically examined by the referring Orthopaedic Surgeon. The pain response was assessed by comparing the scores of the questionnaires. The improvement in clinical examination findings was assessed as well. In patients who underwent facet joint infiltrations, long-term pain improvement was achieved in 79% and in those with nerve blocks in 85%. Immediate pain relief was demonstrated in 83% of patients with nerve infiltrations. No complications were observed. All procedures were very well tolerated by patients. Facet joint and nerve infiltrations under computed tomography guidance constitute an accurate and safe method that could be used to relieve low back pain and minimize the risk of disability.


Idiopathic macrocytic anaemia in the aged: molecular and cytogenetic findings.
Macrocytosis in the elderly is often caused by abnormalities of haematological stem cell differentiation. In this study, a group of elderly patients was analysed for four molecular and cell biological parameters. The aim of the study was to screen elderly patients with idiopathic macrocytic anaemia or MDS for a set of alterations which are related to haematological dysplasia. The analyses used were: DNA-methylation at the calcitonin A gene 5'-area, NRAS point mutations at codons 12 and 13, in vitro colony formation of peripheral blood progenitor cells and cytogenetics of bone marrow cells. The results show that a significant portion of elderly patients with idiopathic macrocytosis have one or more of the abnormalities analysed. Hypermethylation of the calcitonin A gene 5'-area at the chromosome 11 band p15 is relatively common (7/15). Chromosomal aberrations (3/12) and NRAS oncogene point mutations (0/15) were rare findings. In vitro culture of erythroid progenitor cells was relatively frequently abnormal (7/15). Eight of our nine macrocytic patients who did not fulfill the FAB criteria for MDS had at least one of the alterations studied; this suggests that these patients might represent early phases of a stem cell disorder.


Implementing a Novel Software Program to Support Pharmacy Students' Reflective Practice in Scientific Research.
Objective. To explore pharmacy students' perceptions of a novel web application tool (AcaWriter) implemented in a Master of Pharmacy curriculum to support reflective thinking in scientific research.Methods. A qualitative research design involving a 50-minute focus group (n=12) was used. The focus group session was audio-taped, transcribed verbatim, and analyzed thematically using the Braun and Clarke framework.Results. Analysis generated four themes related to AcaWriter's utility in enhancing students' research thinking and capacity. The themes identified included: ease of use to prompt reflection, tangible tool with non-judgmental capacity; benefits for enhancing self and peer reflection on research techniques and group dynamics; benefits of the reflective writing process to enhance research capacity compared with engaging in reflective dialogue; and benefits beyond the writing process: cultivating self-improvement and self-confidence.Conclusion. The findings of this study show that a novel web application implemented within a pharmacy curriculum can assist students' self and peer reflection on a research task. Further research is needed to explore the impact of using this tool and its relationship with academic performance and outcomes.


Tissue and subcellular localization of oligofurostanosides and their specific degrading beta-glucosidase in Dioscorea caucasica Lipsky.
The application of the histochemical technique using the Ehrlich reagent showed that the oligofurostanosides--the steroid glycosides of the furostan series--are localized in idioblasts (special cells--receptacles) of the epidermal layer of Dioscorea caucasica leaves. The activity of oligofurostanoside-specific beta-glucosidase was localized mainly in the membrane fraction of the thylakoides and was also found in the Dioscorea caucasica leaves. We have suggested, that the differential tissue, the subcellular localization of the oligofurostanosides and their degrading enzymes provide the maintenance of the furostanol structure of steroid glycosides which is necessary for their transport within the leaf and from the leaf to the rhizome.


Vehicle Firearm Storage: Prevalence and Correlates in a Sample of Male Firearm Owners.
Previous research has linked unsafe firearm storage practices and other ownership-related characteristics to key factors that facilitate the transition from suicidal thoughts to suicidal behaviors (i.e., acquired capability for suicide). This research has not investigated the extent to which firearm owners store firearms in their vehicles, a factor that increases ready access to the most lethal means of suicide. This cross-sectional study investigated the prevalence of occasional and permanent vehicle firearm storage as well as demographic and psychological correlates of this practice in a sample of N = 408 adult male firearm owners oversampled for historical thoughts of suicide. Participants completed an online survey for monetary compensation. Over 40% of participants indicated at least occasionally storing firearms in their vehicles with over 15% indicating storage of firearms unlocked and loaded. Elevated scores on measures of negative affect, worry, intolerance of uncertainty, and historical suicidal thoughts and behaviors were seen in those who endorsed vehicle firearm storage compared to those who did not. Individual and public health firearm safety strategies would benefit from an explicit focus on the transition of firearms to and from one's vehicle.HIGHLIGHTSOver 40% of male firearm owners at least occasionally store a firearm in a vehicle.15% of those who store a firearm in a vehicle do so unlocked and loaded.Suicidal thought severity was related to storing a firearm in a vehicle.Historical suicidal behaviors were related to storing a firearm in a vehicle.


Source-sink regulation of cotyledonary reserve mobilization during cashew (Anacardium occidentale) seedling establishment under NaCl salinity.
Seedling establishment is a critical process to crop productivity, especially under saline conditions. This work was carried out to investigate the hypothesis that reserve mobilization is coordinated with salt-induced inhibition of seedling growth due to changes in source-sink relations. To test this hypothesis, cashew nuts (Anacardium occidentale) were sown in vermiculite irrigated daily with distilled water (control) or 50mM NaCl and they were evaluated at discrete developmental stages from the seed germination until the whole seedling establishment. The salt treatment coordinately delayed the seedling growth and the cotyledonary reserve mobilization. However, these effects were more pronounced at late seedling establishment than in earlier stages. The storage protein mobilization was affected by salt stress before the lipid and starch breakdown. The globulin fraction represented the most important storage proteins of cashew cotyledons, and its mobilization was markedly delayed by NaCl along the seedling establishment. Free amino acids were mostly retained in the cotyledons of salt-treated seedlings when the mobilization of storage proteins, lipids and starch was strongly delayed. Proline was not considerably accumulated in the cotyledons of cashew seedlings as a response to NaCl salinity. According to these results it is noteworthy that the salt-induced inhibition of seedling growth is narrowly coordinated with the delay of reserve mobilization and the accumulation of hydrolysis products in cotyledons. Also, it was evidenced that free amino acids, especially those related to nitrogen transport, are potential signals involved in the regulation of storage protein hydrolysis during cashew seedling establishment under NaCl salinity.


Association of PNPLA3 rs738409 G/C gene polymorphism with nonalcoholic fatty liver disease in children: a meta-analysis.
Nonalcoholic fatty liver disease (NAFLD) is one of the most common causes of chronic liver disease worldwide. Current studies have shown that PNPLA3 (Patatin-like phospholipase domain containing 3) rs738409 G/C gene polymorphism is associated with adult nonalcoholic fatty liver disease [1, 2].But there is no consensus on the relationship between PNPLA3 rs738409 G/C gene polymorphism and children NAFLD due to differences in population samples. To this end, a meta-analysis of published research is conducted to comprehensively assess the relationship between PNPLA3 gene polymorphism and NAFLD in children. We searched MEDLINE, PubMed, EMBASE, and CENTRAL databases from inception to May 2019. Case-control studies assessing the relationship between PNPLA3 rs738409 G/C gene polymorphism with non-alcoholic fatty liver disease in children were selected according to inclusion and exclusion criteria. Random effects model was used to quantify the association between the PNPLA3 rs738409 G/C gene polymorphism and the susceptibility of children's NAFLD. Fixed effects model was used to quantify the relationship between the PNPLA3 rs738409 G/C gene polymorphism and the severity of NAFLD in children. The Stata 12.0 software was employed for data analysis. A total of nine case-control studies were included in this meta-analysis containing data of 1173 children with NAFLD and 1792 healthy controls. Five studies compared NAFLD children and non-NAFLD healthy populations. Statistical analysis showed that PNPLA3 gene polymorphism was significantly associated with children's NAFLD in the allele contrast, dominant, recessive and over dominant models (G vs C,OR = 3.343, 95% CI = 1.524-7.334; GG + GC vs CC,OR = 3.157, 95% CI = 1.446-6.892;GG vs GC + CC,OR = 5.692, 95% CI = 1.941-16.689; GG + CC vs GC,OR = 2.756, 95% CI = 1.729-4.392). Four case-control studies compared Children with nonalcoholic fatty liver (NAFL) and children with nonalcoholic steatohepatitis (NASH). The results showed that the PNPLA3 gene polymorphism was also significantly associated with the severity of NAFLD in children in recessive gene model (GG vs GC + CC,OR = 14.43, 95% CI = 5.985-34.997); The Egger's test revealed no significant publication bias. Meta-analysis showed that PNPLA3 gene polymorphism was significantly associated with susceptibility and severity of NAFLD in children.


Measurement of circulating cell-free DNA in relation to 18F-fluorocholine PET/CT imaging in chemotherapy-treated advanced prostate cancer.
To examine the effects of chemotherapy on circulating cell-free DNA (cfDNA) composition in relation to investigational whole-body measurement of tumor activity by fluorine-18 fluorocholine (FCH) positron emission tomography/computed tomography (PET/CT) in hormone-refractory prostate cancer (HRPC). Serial FCH PET/CT scans were performed in eight patients with HRPC receiving docetaxel-based chemotherapy. Corresponding serial cfDNA samples were characterized by microfluidic electrophoresis, quantified by real-time PCR, and compared with PET/CT results. Promoter methylation of two prostate cancer-associated genes, GSTP1 and RARB2, was assessed by methylation-specific PCR of bisulfite-converted cfDNA. Plasma cfDNA concentrations increased significantly from 13.3 ng/mL at baseline to 46.8 ng/mL and 50.9 ng/mL after one and three treatment cycles, respectively (p= 0.001). GSTP1 and/or RARB2 promoter methylation was identified in all pretreatment samples. The appearance of large (200 bp-10.4 kb) cfDNA fragments was noted in posttreatment samples along with loss of methylation at GSTP1 and/or RARB2. Tumor activity on PET/CT correlated with cfDNA concentration (r=-0.50, p= 0.01). Patients meeting criteria for PET tumor response had significantly lower pretreatment cfDNA levels than those who did not (8.0 vs. 16.4 ng/mL, p= 0.03). Chemotherapy is associated with significant changes in plasma cfDNA content and FCH PET/CT-detected tumor activity. These interrelated measures are potential candidate markers of therapeutic response in HRPC.


Insights into lactic acid bacteria cryoresistance using FTIR microspectroscopy.
Freezing is widely used for bacterial cell preservation. However, resistance to freezing can greatly vary depending on bacterial species or growth conditions. Our study aims at identifying cellular markers of cryoresistance based on the comparison of three lactic acid bacteria (LAB) exhibiting different tolerance to freezing: Carnobacterium maltaromaticum CNCM I-3298, Lactobacillus delbrueckii subsp. bulgaricus ATCC 11842, and Lactobacillus delbrueckii subsp. bulgaricus CFL1. A thorough characterization of their cytoplasmic membrane properties was carried out by measuring their fatty acid composition, membrane fluidity, and lipid phase transition upon cooling from 50 to -50 °C. Vitrification temperatures of the intra- and extra-cellular compartments were also quantified by differential scanning calorimetry. Additionally, the cell biochemical characterization was carried out using a recently developed Fourier transform infrared (FTIR) micro-spectroscopic approach allowing the analysis of live bacteria in an aqueous environment. The multivariate analysis of the FTIR spectra of fresh and thawed cells enabled the discrimination of the three bacteria according to their lipid, protein, and cell wall peptidoglycan components. It also revealed freezing-induced modifications of these three cellular components and an increase in bacteria heterogeneity for the two strains of L. bulgaricus, the freeze-sensitive bacteria. No cellular damage was observed for C. maltaromaticum, the freeze-resistant bacteria. Comparison of the results obtained from the different analytical methods confirmed previously reported cryoresistance markers and suggested new ones, such as changes in the absorbance of specific infrared spectral bands. FTIR microspectroscopy could be used as a rapid and non-invasive technique to evaluate the freeze-sensitivity of LAB.


Dysbiotic gut microbiome: A key element of Crohn's disease.
Since the first publication on "regional ileitis", the relevance of this chronic inflammatory disease condition termed finally as Crohn's disease is continuously increasing. Although we are beginning to comprehend certain aspects of its pathogenesis, many facets remain unexplored. Host's gut microbiota is involved in a wide range of physiological and pathological processes including immune system development, and pathogen regulation. Further, the microbiome is thought to play a key role in Crohn's disease. The presence of Crohn's-associated variants of NOD2 and ATG16L genes appears to be associated not only with alterations of mucosal barrier functions, and bacterial killing, but the gut microbiota, as well, reflecting a potential relationship between the host's genotype and intestinal dysbiosis, involved in disease etiology. This review aims to characterize some exciting new aspect of Crohn's disease pathology, focusing mainly on the role of intestinal microbes, and their interplay with the immune system of the host.


Computational Study on Homolytic Bond Energies of the Ag-X (X = C, O, and H) Complexes and Hammett-Type Analysis of Reactivity.
Thirty-seven calculation methods were benchmarked against the available experimental bond lengths and energies data regarding the Ag-X bonds. The theoretical protocol PBE0/VDZ//ωB97x-D/mVTZ was found to be capable of accurately predicting the homolytic bond dissociation energies (BDEs) of Ag-X complexes with a precision of 1.9 kcal/mol. With the available method in hand, a wide range of different Ag-X BDEs were estimated. BDE(Ag-CH2X), BDE(Ag-PhX), BDE(Ag-OPhX), and BDE(Ag-OCOPhX) (X = NH2, OMe, Me, H, Cl, and NO2) were found to be in the ranges of 27-47, 51-54, 19-39, and 64-70 kcal/mol, respectively. Subsequently, Hammett-type analysis was carried out with reactivity parameters. Good positive linear relationships were found for BDE of Ag-O bands and decarboxylation barriers of Ag-OCOPhX with the Hammett constant σ. It suggested that electron-donating substituents could promote either the homolytic cleavage of the Ag-OPhX bond to undergo a radical process or Ag-OCOPhX decarboxylation. Moreover, ligand effects on Ag-H bonds were investigated using BDE(Ag-H) and related NPA charges on Ag. In the case of P-ligands, carbene ligands, and other small molecule ligands (i.e., CO, CO2, and H2O), a good negative linear relationship was found. In contrast, N-ligands could have a reverse effect. Understanding the intrinsic relationships of BDE(Ag-X) with related reactivity parameters might help gain insights into the structure-reactivity relationships in Ag-X-assisted C-H activation/decarboxylation.


Computed tomographic enterography (CTE) in evaluating bowel involvement in patients with ovarian cancer.
To explore the utility of CTE in the evaluation of bowel invasion in patients with primary ovarian, fallopian tube, and peritoneal cancer. This observational study included 73 patients who received CTE before operation between September 2019 and December 2021. Two radiologists reviewed CTE images, focusing on the sites and depth of bowel involvement. Based on the findings during surgical exploration, we evaluated the diagnostic power, like sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (+ LR), and negative likelihood ratio (- LR) of CTE. Additionally, the characteristic images of bowel involvement on CTE corresponding to surgical findings were shown in the study. The rate of macroscopic bowel invasion in this cohort was 49.31% (36/73), of which eight patients had small bowel involvement, 17 patients had colon involvement and 27 patients had sigmoid-rectum involvement. CTE detected bowel invasion in the small intestine with a sensitivity, specificity, PPV, NPV, and accuracy of 87.50%, 92.31%, 58.33%, 98.36%, 91.78%; for colon, the statistics were 58.82%, 96.43%, 83.33%, 88.52%, 87.67% and for sigmoid-rectum 62.96%, 82.61%, 68.00%, 79.17%, 75.34%, respectively. CTE appeared a preferable diagnostic power on the small bowel and colon invasion in patients with primary ovarian, fallopian tube, and peritoneal cancer.


Immunotherapy for Esophageal Cancer: State-of-the Art in 2021.
The management of esophageal cancer (EC) has experienced manifold changes during the last decades. Centralization of EC treatment has been introduced in many countries, subsequently allowing the development of specialized high-volume centers. Minimal invasive surgery has replaced open surgery in many centers, whereas more potent systemic treatments have been introduced in clinical practice. Newer chemotherapy regimens increase long-term survival. Nevertheless, the overall survival of EC patients remains dismal for advanced tumor stages. In this direction, a wide range of targeted biologic agents (immunotherapy) is currently under assessment. Anti- Human Epidermal Growth Factor Receptor-2 (HER-2) monoclonal antibodies are used in HER2 (+) tumors, predominantly well-differentiated adenocarcinomas, and are currently assessed in the neoadjuvant setting (TRAP, INNOVATION trials). Immune checkpoint inhibitors Nivolumab (ATTRACTION-03) and pembrolizumab (KEYNOTE-181), have demonstrated a survival benefit compared with conventional chemotherapy in heavily pre-treated progressive disease. More recently, CheckMate-577 showed very promising results for nivolumab in a curative adjuvant setting, improving disease-free survival mainly for esophageal squamous cell carcinoma. Several ongoing trials are investigating novel targeted agents in the preoperative setting of locally advanced EC. In addition, other immunomodulatory approaches such as peptide vaccines and tumor infiltrating lymphocytes (TILs) are currently under development and should be increasingly integrated into clinical practice.


Electron microscopic and immunohistochemical studies on Hashimoto's thyroiditis.
Three types of Hashimoto's thyroiditis, lymphoid type (L-type), oxyphilic epithelium (O-type) and pronounced epithelial destruction (p-type), were ultrastructurally and immunohistochemically studied. In both the L-type and the O-type, two different kinds of follicles, active and degenerated ones, were ultrastructurally found. Although the follicular conformation was generally conserved, focal basement membrane damage was frequently found around the generated follicles. Some plasma cells and lymphocytes transversed the basement membranes and could be seen in the spaces between adjacent epithelial cells. In the P-type, most epithelial cells were replaced by fibrillar fibrous bundles, and destructive products probably derived from epithelial cells were scattered in the stroma. Immunoglobulin (IgG, IgA and IgM) deposittions was found not only in some basement membranes, colloid, epithelial cells and peeled-off cells in the follicles but also in the stroma. Some of the infiltrating plasma cells and lymphocytes had immunoglobulins in the cytoplasm. The intensity o the positive staining was by far the strongest for IgG, and the number of immunoglobulin-containing immunocytes was also the largest for IgG.


Wound contraction decreases with intravenously injected substance P in rabbits.
Substance P is an injury-inducible endogenous factor for the mobilization of CD29+ stromal-like cells into circulation and that are major effectors of accelerated healing. In this study, we evaluated the effect of intravenously injected substance P on full-thickness skin wound healing as a secondary intention wound model. We made circular full-thickness skin wounds on the ears of 28 New Zealand white rabbits. They were treated with phosphate-buffered saline, or intravenous 5, 50, or 250 n mole/kg substance P at days 0 and 1. All substance P-treated groups showed a 2.6-5.4-fold higher CD29 expression and resulted in greatly decreased wound contraction and early maturation of the stroma. However, a significant decrease in wound contraction was measured only in the 5 n mole/kg treatment group. We conclude that intravenously injected substance P at 5 n mole/kg decreases wound contraction and promotes wound maturation in full-thickness skin wounds in a rabbit ear model.


Fine root production of astringent phenolics.
Chemical quality of fine roots (<1 mm diameter) was determined over a gradient of species composition in the Mixed Mesophytic Forest Region. Ash-free nitrogen, calcium, and phosphorus concentrations of roots declined by 49, 41, and 72%, respectively, over a gradient of increasing soil acidity (pH 5.3 to 4.7). Lignin concentration was unrelated to either the vegetation gradient or any of the soil changes it encompassed; however, astringent phenolics increased by 275% over the same gradient. Trends in the chemical constituency of fine roots suggest that the production of phenolics in below-ground plant parts is increased on nutrient-poor sites. This response is best related to changes in species composition, especially increasing importancy of Quercus spp.


Improved process for the production of cellulose sulfate using sulfuric acid/ethanol solution.
An improved process for production of cellulose sulfate (CS) was developed by using sulfuric acid/ethanol solution as sulfonating agent and Na2SO4 as water absorbent. The FTIR, SEM and TG analysis were used to characterize the CS prepared. The total degree of substitution and viscosity of the product solution (2%, w/v) were ranging from 0.28 to 0.77 and from 115 to 907 mPa s, respectively, by changing the process parameters such as the amount of Na2SO4, the reaction time, the temperature, the sulfuric acid/alcohol ratio and liquid/solid ratio. The results indicated that the product with DS (0.28-0.77) and η2% (115-907) mPa s could be produced by using this improved process and more cellulose sulfate could be produced when cellulose was sulfonated for 3-4 h at -2 °C in sulfuric acid/ethanol (1.4-1.6) solution with addition of 0.8 g Na2SO4. The (13)C NMR indicated that the sulfate group of CS produced using sulfuric acid/ethanol solution was at C6 position.