Telomerase deficiency impairs differentiation of mesenchymal stem cells.
Expression of telomerase activity presumably is involved in maintaining self-replication and the undifferentiated state of stem cells. Adult mouse bone marrow mesenchymal stem cells (mMSCs) are multipotential cells capable of differentiating into a variety of lineage cell types, including adipocytes and chondrocytes. Here we show that the lacking telomerase of mMSC lose multipotency and the capacity to differentiate. Primary cultures of mMSCs were obtained from both telomerase knockout (mTR(-/-)) and wild-type (WT) mice. The MSCs isolated from mTR(-/-) mice failed to differentiate into adipocytes and chondrocytes, even at early passages, whereas WT MSCs were capable of differentiation. Consistent with other cell types, late passages mTR(-/-)MSCs underwent senescence and were accompanied by telomere loss and chromosomal end-to-end fusions. These results suggest that in addition to its known role in cell replication, telomerase is required for differentiation of mMSCs in vitro. This work may be significant for further potentiating adult stem cells for use in tissue engineering and gene therapy and for understanding the significance of telomerase expression in the process of cell differentiation.


Single-agent chemotherapy for advanced adenocarcinoma of the lung. A review.
Systemic therapy with cytostatic agents has been widely used in the management of inoperable adenocarcinoma of the lung (ACL). However, chemotherapy for this tumor type remains experimental, and the prognosis is still poor. Thus, the literature on single-agent chemotherapy was reviewed in order to establish critical background material for the planning and evaluation of future studies. Only vindesine, dibromodulcitol, doxorubicin and hexamethylmelamine have displayed overall response rates exceeding 10% in randomized studies. Several of the most promising agents with response rates above 20% in non-randomized studies, i.e., 5-fluorouracil, mitomycin C, vinblastine and ifosfamide, have not been adequately evaluated in randomized trials in ACL. There is no published evidence to suggest the superiority of single-agent chemotherapy over the best supportive treatment, with respect either to survival or to quality of life. There are considerable methodological problems in designing, executing, analyzing and reporting these studies. Some of the problems could be solved by use of the internationally accepted guidelines for reporting results of cancer treatment, which might make more rapid progress possible.


Association between HLA alleles and sub-phenotype of childhood steroid-sensitive nephrotic syndrome.
Few studies have addressed the effects of human leukocyte antigen (HLA) alleles on different clinical sub-phenotypes in childhood steroid-sensitive nephrotic syndrome (SSNS), including SSNS without recurrence (SSNSWR) and steroid-dependent nephrotic syndrome/frequently relapse nephrotic syndrome (SDNS/FRNS). In this study, we investigated the relationship between HLA system and children with SSNSWR and SDNS/FRNS and clarified the value of HLA allele detection for precise typing of childhood SSNS. A total of 241 Chinese Han individuals with SSNS were genotyped using GenCap-WES Capture Kit, and four-digit resolution HLA alleles were imputed from available Genome Wide Association data. The distribution and carrying frequency of HLA alleles in SSNSWR and SDNS/FRNS were investigated. Additionally, logistic regression and mediating effects were used to examine the relationship between risk factors for disease process and HLA system. Compared with SSNSWR, significantly decreased serum levels of complement 3 (C3) and complement 4 (C4) at onset were detected in SDNS/FRNS (C3, P < 0.001; C4, P = 0.018). The average time to remission after sufficient initial steroid treatment in SDNS/FRNS was significantly longer than that in SSNSWR (P = 0.0001). Low level of C4 was further identified as an independent risk factor for SDNS/FRNS (P = 0.008, odds ratio = 0.174, 95% confidence interval 0.048-0.630). The HLA-A*11:01 allele was independently associated with SSNSWR and SDNS/FRNS (P = 0.0012 and P = 0.0006, respectively). No significant HLA alleles were detected between SSNSWR and SDNS/FRNS. In addition, a mediating effect among HLA-I alleles (HLA-B*15:11, HLA-B*44:03 and HLA-C*07:06), C4 level and SDNS/FRNS was identified. HLA-I alleles provide novel genetic markers for SSNSWR and SDNS/FRNS. HLA-I antigens may be involved in steroid dependent or frequent relapse in children with SSNS as mediators of immunoregulation.


Histamine stimulates inositol phosphate accumulation via the H1-receptor in cultured human endothelial cells.
The effects of histamine on [3H]inositol phosphate ([3H]IP) accumulation was examined in the presence of lithium in [3H]inositol-prelabelled human umbilical vein endothelial cells. Histamine stimulated total [3H]IP formation in a dose-dependent manner with a half-maximal value (EC50) of around 1-2 X 10(-6) M. Mepyramine, but not cimetidine, completely abolished the histamine response indicating that activation of phosphoinositide hydrolysis is mediated via H1-receptors. These data are the first to suggest that activation of inositol lipid hydrolysis is the underlying transmembrane signalling mechanism histamine H1-receptors employ in mediating various endothelial cell functions.


Osteogenic periosteum esterase activity: a comparative morphological and cytochemical study of bone cells in situ on rat proximal tibiae and in smears.
Mononucleate phagocytes are presently thought to participate, along with osteoclasts, in bone resorption. Similar cells may also be precursors to osteoclasts. Esterase is a hydrolytic enzyme commonly found in most phagocytes and cells such as these have been shown to degrade bone matrix in vitro. Therefore, periosteal cells of remodeling proximal tibiae in rats were examined for esterase activity. Stain reactions were compared both in situ and in smears. Percentages of different cell types were obtained by counting cells in Wright-Giemsa-stained smears. Osteoclasts and mononucleate phagocyte-like cells made up approximately 20% of the isolated cells. Cells resembling preosteoblasts, mononucleate phagocytes, and osteoblasts stained for esterase activity with light to moderate intensity. Osteoclasts and mononucleate osteoclast-like cells were strongly esterase positive. The esterase was characterized as being type B by using nonorganophosphorus inhibitors of the various esterase types. The addition of esterase to the list of enzyme activities of cells in close proximity to osteoclasts lends further support to the hypothesis that mononucleate cells also participate in bone resorption.


Sustained release of bovine serum albumin using implantable wafers prepared by MPEG-PLGA diblock copolymers.
MPEG-PLGA diblock copolymers, consisting of methoxy polyethylene glycol (MPEG) and poly(L-lactic-co-glycolic acid) (PLGA), were synthesized by ring-opening polymerization of L-lactide and glycolide in the presence of MPEG as an initiator. Implantable wafers, using diblock copolymers as a drug carrier, were fabricated by direct compression method after freeze milling of the diblock copolymers and bovine serum albumin-fluorescein isothiocyanate (BSA-FITC) as a model protein drug. The wafers prepared with MPEG-PLGA diblock copolymers exhibited initial burst in the release of BSA. The BSA release profiles from the wafers depended on MPEG-PLGA diblock copolymer compositions. The in vitro release of the BSA also correlated with the degradation rate of the PLGA part in the diblock polymers. The wafers prepared from diblock copolymers with an increased MPEG segment showed the more structural metamorphosis of crack form due to higher water absorption of MPEG inside the wafer, and induced faster BSA release. The wafers prepared by using MPEG-PLGA diblock copolymers in the presence of small intestinal submucosa (SIS) as a drug carrier additive exhibited controlled BSA release profiles, although the wafers exhibited release patterns with a lag time at the initial stage as the MPEG segment in diblock copolymer compositions increased. Thus, we confirmed that the MPEG-PLGA diblock copolymers could be used as a protein delivery carrier in implantable wafer form.


Comparing HMME-PDT and Cynergy dual-wavelength laser in the treatment of facial PWS.
Pulsed dye laser (PDL) is the gold-standard therapy for port-wine stains (PWSs), but the therapeutic efficacies vary widely with each individual. Recent studies suggest that hematoporphyrin monomethyl ether-photodynamic therapy (HMME-PDT) is an effective alternative in the treatment of PWS. This retrospective study aimed to compare the effects of HMME-PDT and Cynergy dual-wavelength laser on red and purple color types of PWS METHODS: This retrospective study included 38 PWS patients who received Cynergy dual-wavelength laser treatment (PDL group) and 39 PWS patients who received HMME-PDT treatment (HMME-PDT group). Patients in each group received two treatments. The curative effect of purple or red PWS in two groups was analyzed. After the first treatment, the total significant response rate and the total response rate of the HMME-PDT group were 43.59% and 74.36%, which were significantly higher than those of the PDL group (5.26% and 47.36%) (P < 0.05). After the second treatment, the total significant response rate and the total response rate of the HMME-PDT group were 69.23% and 100%, which were obviously higher than those of the PDL group (34.21% and 68.42%) (P < 0.05). For both groups, the total significant response rate of purple-type PWS was remarkably higher than that of red-type PWS (the HMME-PDT group: P = 0.004; the PDL group: P = 0.048). There was no obvious difference in terms of patient age and PWS location between the HMME-PDT and PDL groups. Furthermore, there was no considerable difference in the incidence of adverse reactions between the two groups (P > 0.05). The single-center retrospective study suggested that HMME-PDT might have better efficacies on purple and red PWS than PDL treatment. The total treatment efficacy was greater in purple-type PWS than in red-type PWS.


Full-length infectious clone of an Iranian isolate of chicken anemia virus.
An Iranian field strain of chicken anemia virus (CAV), designated IR CAV, was isolated in the Marek's disease virus-transformed lymphoblastoid cell line MDCC-MSB1 (MSB1) culture for the first time. The full-length CAV DNA of this strain was cloned in the bacterial plasmid pTZ57R/T to create the molecular clone pTZ-CAV. The nucleotide and deduced amino acid sequences of viral proteins of IR CAV were compared with those of representative CAV sequences including reference and commercial vaccine strains. IR CAV was not related to vaccine strains and also found to have glutamine at positions 139 and 144 confirming previous studies in which such mutations were associated with a slow rate of virus spread in cell culture. pTZ-CAV was digested with PstI to release IR CAV DNA and then transfected into MSB1 cell by electroporation. The transfected cells showed cytopathic effect similar to virion-initiated infection. One-day old specific pathogen-free chicks were inoculated with the regenerated virus, which had been obtained from transfected MSB1 cells, and compared with the chicks inoculated with IR CAV. Gross lesions in the birds inoculated with the regenerated virus illustrated the infectious nature of the regenerated virus from the cloned IR CAV DNA.


Effects of curcumin on growth performance, jejunal mucosal membrane integrity, morphology and immune status in weaned piglets challenged with enterotoxigenic Escherichia coli.
The aim of this study was to evaluate the effect of dietary curcumin supplementation on growth performance and intestinal mucosal barrier function of weaned piglets. Fifty piglets, weaned at 21±2 days of age, were randomly allotted to five treatments for 21 days. The dietary treatments were the control (basal diet), and the basal diet supplemented with 50mg/kg quinocetone, or 200 mg/kg, 300 mg/kg or 400mg/kg curcumin. The piglets were housed in individual pens and orally challenged with enterotoxigenic Escherichia coli (ETEC) during the preliminary trial period. The jejunal morphology and histology analysis were detected under light microscope. The plasma D-lactate and diamine oxidase (DAO) were determined by using enzymatic spectrophotometric assay. Immunohistochemistry assays were used to examine secretory immunoglobulin (sIgA) protein expression. Real-time PCR was used to determine mRNA levels of cytokine and Toll-like receptor 4 (TLR4) in jejunal mucosa. The results showed that, compared with the control, dietary addition of 300 mg/kg or 400 mg/kg curcumin decreased (P<0.05) feed/gain ratio and crypt depth, improved (P<0.05) villus height and villus height:crypt depth ratio, reduced (P<0.05) plasma D-lactate and DAO activity, up-regulated the protein expression of sIgA (P<0.05), increased (P<0.05) the number of goblet cells (GCs) and reduced (P<0.05) the number of intraepithelial lymphocytes (IELs). The mRNA levels of interleukin 1β (IL-1β) and TLR4 and tumor necrosis factor α (TNF-α) were also decreased (P<0.05), but mRNA level of interleukin 10 (IL-10) was increased (P<0.05). There was no difference in the above parameters between the 300 mg/kg and 400 mg/kg curcumin groups. Pigs fed with 50 mg/kg quinocetone also decreased (P<0.05) feed/gain ratio, increased villus height:crypt depth ratio (P<0.05), and reduced (P<0.05) crypt depth and mRNA levels of TLR4. In conclusion, curcumin and the quinocetone have similar effects in improving piglet growth, but dietary addition of 300 mg/kg or 400 mg/kg curcumin was more effective than quinocetone in improving intestinal mucosal barrier integrity, morphology, and immune status of weaned pigs. This indicates that curcumin could be used as a potential feed additive replacing quinocetone in weaned piglets.


Aneurysms of saphenous vein grafts as late complication of coronary artery bypass surgery: successful exclusion by percutaneous transcatheter embolization.
We report a case of a 71-year-old man with a long coronary artery disease history and two sets of coronary artery bypass grafts. He developed large aortocoronary saphenous vein graft aneurysms in the two grafts from the first set of aortocoronary saphenous venous 20 years later. During the previous 3 years, the aneurysms grew in diameter from 22 to 50 mm. Because of severe renal insufficiency, and in order to avoid jeopardizing the late normal coronary artery bypass grafts by further thoracic surgery, we excluded successfully these altered grafts percutaneously by using 13 coils during the same procedure.


Unilateral bi-portal endoscopy for unilateral L5 nerve injury due to post-traumatic epidural hematoma at T12/L1 level: A case report and literature review.
Spinal epidural hematoma (SEH) is an uncommon condition that can result in severe neurological problems and needs to be treated as soon as possible. The incidence of traumatic SEH is 0.5 %-1.7 %, but increases to 9 % in patients with rheumatic diseases. Surgical treatment options include open surgery and minimally invasive surgery. We reported a post-traumatic SEH at T12/L1 level combined with L5 nerve injury and treated by UBE technique. To our knowledge, there was no reported cases like this. A 38-year-old man with left leg weakness and severe back pain after fell down while cycling. Physical examination suggested left hip abduction was 2/5 strength, left dorsiflexion of hallux dorsal extension was 0/5 strength and the left ankle dorsiflexion was 2/5 strength. Magnetic resonance images (MRI) of lumbar spine showed a two-leveled hematoma extending from T12 to L1. After 1 year of surgery, the patient's symptoms had largely disappeared and he was able to perform daily activities independently. An epidural hematoma at the L1 level is can cause symptoms of the L5 nerve root alone, which may be due to anatomical reasons. Complete removal of the epidural hematoma is necessary to restore the function of the nerve. We report a case of successful removal of an epidural hematoma using the UBE technique with good postoperative results. The single nerve injury can occur with a thoracolumbar segmental hematoma, and UBE technology could be used to remove epidural hematoma.


A randomized comparison of extended-release naltrexone with or without patient navigation vs enhanced treatment-as-usual for incarcerated adults with opioid use disorder.
The high prevalence of opioid use among justice-involved adults make jails an exceptional setting to initiate opioid use disorder (OUD) treatment, but optimal strategies for delivering these interventions are still not well understood. The objective of this study was to conduct a randomized controlled trial to assess the effectiveness of extended-release naltrexone (XR-NTX, Vivitrol®; Alkermes Inc) alone or in conjunction with patient navigation (XR-NTX + PN) for jail inmates with OUD. We randomized a sample of 135 sentenced jail inmates with moderate to severe OUD to (1) XR-NTX only; (2) XR-NTX + PN; or (3) enhanced treatment-as-usual (ETAU) with drug education, each initiated prior to release from jail. We scheduled follow-up data assessments at 1, 3, 6, and 12 months post-release. Primary outcomes were opioid use (based on Timeline Followback Interview and Addiction Severity Index) and meeting CIDI DSM-5 criteria for OUD 6 months postrelease. We also measured treatment adherence, HIV risk, and recidivism. XR-NTX participants received a mean of 2.26 of 7 possible injections compared to XR-NTX + PN participants, who received a mean of 2.93 injections (Cohen's d = 0.33, 95% CI: -0.09 to 0.74). Thirty-six percent of patients in XR-NTX + PN attended at least one postrelease PN session. We found no significant differences by study condition six months after release from jail for the primary outcomes of any opioid use (ETAU: 17%, XR-NTX: 16%, XR-NTX + PN: 29%) and past 30-day OUD (ETAU: 8%, XR-NTX: 11%, XR-NTX + PN: 10%). Secondary outcomes of rearrest and HIV risk also were similar across groups, with the exception of lower sex-related HIV risk among those in the XR-NTX condition at 12 months. This study did not show superior outcomes of XR-NTX or XR-NTX + PN with regard to opioid use or recidivism outcomes, relative to ETAU. It did, however, highlight the difficulties with adherence to XR-NTX and PN interventions in OUD patients initiating treatment in jail.


Lifetime distribution of clusters in binary mixtures involving hydrogen bonding liquids.
Hydrogen bonded liquids are associated liquids and tend to exhibit local inhomogeneity in the form of clusters and segregated sub-nano domains. It is an open question as to whether Hbonded clusters in pure water have common features with the water segregated pockets observed in various aqueous binary mixtures, such as water-alcohol mixtures, for example. In the present study, we demonstrate through classical molecular dynamics studies of the lifetime distributions of the hydrogen bonds in different types of binary mixtures, that these lifetimes exhibit the same universal features in the case of the pure liquids, independently of the species concentrations. The same types of three distinct lifetimes are observed, all of them in the sub picosecond regime. The primary lifetime concerns that of Hbonded dimers, and strongly depends on Hbonding criteria such as the bonding distance. The two others are independent of bonding criteria and appear as universal accross many liquids and mixtures. The secondary lifetime ([Formula: see text] fs) concerns Hbonded cluster lifetimes, while the tertiary lifetime ([Formula: see text] fs) concerns the topology of these clusters, such as chains or globules, for example. This surprizing separation in three distinct lifetimes suggests the existence of associated three distinct kinetic mechanisms in the very short sub-picosecond time scales, with, in addition, an appealing connection to the concepts of local energy and entropy.


Synthesis and antibacterial activity of O-methyl derivatives of azalide antibiotics: II. 6-OME derivatives via clarithromycin.
Direct O-methylation of various derivatives of 9-deoxo-8a- and 9a-aza-8a-homo-erythromycin (2',3'-bis-Cbz protected) gives 6-OMe derivatives only in a small number of special cases. The 6-OMe-azalides can, however, be synthesized beginning from clarithromycin.


Cervical cerclage application algorithm in continued cervical shortening cases despite vaginal progesterone.
The aim of this study is evaluation of pregnancy outcomes of the asymptomatic cases with vaginal progesterone treatment for the 20-30 mm cervical length detected in the transvaginal ultrasonography for fetal abnormality screening and cervical cerclage after cervical length detected <20 mm in weekly cervical length measures; and present the treatment algorithm of progesterone treatment combined with cervical cerclage application. Patients who have the inclusion criteria and cervical length more than 30 mm were categorized as group 1(n = 1948). Group 2 were included patients with cervical length shorter than 30 mm (n = 95). All patients of group 2 started to use vaginal natural progesterone 400 mg/day(n = 87). Pregnancies which progressed with cervical length above 20 mm were continued vaginal progesterone until 34. Gestational week and they were named as group 2A (n = 78). Cervical cerclage were applied to patients with cervical length below than 20 mm measured via transvaginal ultrasonography and they were categorized as group 2B (n = 9). Excluding 8 cases (8.42 %) which cervical cerclage were applied, 87 patients were administered vaginal progesterone and in 9 cases (10.34 %) cervical shortening were continued despite progesterone treatment. Four out of these nine cases (44.44 %) had cervical cerclage and their deliveries were delayed after 34 th gestational week. Vaginal progesterone treatment prevented cervical shortening in 89.66 % of patients who had cervical length between 20-30 mm and out of these patients. Cervical cerclage application algorithm in continued cervical shortening cases despite vaginal progesterone is beneficial to delay the delivery after 34 th gestational week and related to low complication rate.


A novel simplified method for extraction of microplastic particles from face scrub and laundry wastewater.
Microplastic pollution in different environmental matrices is a serious concern in the recent times. Personal care products and washing of synthetic fabrics are some of the main sources of microplastic pollution. In this work, a novel simplified, effective and sustainable method for extraction of microplastic particles from face scrub and laundry wastewater was developed. Different parameters affecting the extraction were analysed and the extraction process was optimised. The extraction efficiency of the proposed method was found to be ~ 94.1 ± 1.65%, which was slightly better than the previously available method with an advantage of ease in extraction and lesser time and resource consuming. The developed method was used to demonstrate the extraction of microplastic particles from 12 face scrub samples with different brands. It was found that the samples contained microplastic particles of varying size. The physical and chemical structure intactness of microplastic particles during the extraction was also analysed and found to be acceptable. The developed extraction method was also applied for the extraction of microfibers from the laundry wastewater. It was found that this proposed method is suitable to make the cleaner extracted samples for an easy and more effective qualitative and quantitative analysis of MPs.


Enalapril suppresses ventricular remodeling more effectively than losartan in patients with acute myocardial infarction.
Ventricular remodeling after acute myocardial infarction (AMI) is associated with increased morbidity and mortality. ELITE II study showed that losartan, an angiotensin receptor blocker, shows a survival benefit to the same degree as captopril, an angiotensin-converting enzyme inhibitor, does in patients with heart failure. However, recent OPTIMAAL study showed that clinical outcomes after losartan are not superior to those after captopril in patients with AMI. We examined the effect of losartan on ventricular remodeling after AMI comparatively with that of enalapril. We enrolled 203 consecutive patients with AMI (mean age 62 +/- 11 years). All patients underwent primary percutaneous coronary intervention and were randomly assigned to losartan (25-50 mg, n = 101) or enalapril (2.5-10 mg, n = 102) treatment. Biplane left ventriculography was performed just after primary percutaneous transluminal coronary angioplasty (acute phase) and 6 months after the onset of AMI. Any of the maximal creatine kinase level, left ventricular end-diastolic volume index, end-systolic volume index, and ejection fraction measured at acute phase was not different between losartan and enalapril groups. However, changes in left ventricular end-diastolic index (18 +/- 25 vs 8 +/- 24 mL/m2) and left ventricular end-systolic volume index (10 +/- 20 vs 2 +/- 18 mL/m2) from acute phase to 6 months were significantly greater in losartan than in enalapril group. Change in left ventricular ejection fraction (0.2% +/- 10.3% vs 3.4% +/- 11.6%) from acute phase to 6 months was significantly smaller in losartan than in enalapril group. The plasma level of brain natriuretic peptide at 6 months was significantly higher in losartan than in enalapril group (all P < .05). These indicate that enalapril suppresses ventricular remodeling after AMI more effectively than losartan.


Viral anti-inflammatory serpin reduces immuno-coagulopathic pathology in SARS-CoV-2 mouse models of infection.
SARS-CoV-2 acute respiratory distress syndrome (ARDS) induces uncontrolled lung inflammation and coagulopathy with high mortality. Anti-viral drugs and monoclonal antibodies reduce early COVID-19 severity, but treatments for late-stage immuno-thrombotic syndromes and long COVID are limited. Serine protease inhibitors (SERPINS) regulate activated proteases. The myxoma virus-derived Serp-1 protein is a secreted immunomodulatory serpin that targets activated thrombotic, thrombolytic, and complement proteases as a self-defense strategy to combat clearance. Serp-1 is effective in multiple animal models of inflammatory lung disease and vasculitis. Here, we describe systemic treatment with purified PEGylated Serp-1 as a therapy for immuno-coagulopathic complications during ARDS. Treatment with PEGSerp-1 in two mouse-adapted SARS-CoV-2 models in C57Bl/6 and BALB/c mice reduced lung and heart inflammation, with improved outcomes. PEGSerp-1 significantly reduced M1 macrophages in the lung and heart by modifying urokinase-type plasminogen activator receptor (uPAR), thrombotic proteases, and complement membrane attack complex (MAC). Sequential changes in gene expression for uPAR and serpins (complement and plasminogen inhibitors) were observed. PEGSerp-1 is a highly effective immune-modulator with therapeutic potential for severe viral ARDS, immuno-coagulopathic responses, and Long COVID.


Initial in vitro interaction of osteoblasts with nano-porous alumina.
In the present study we have used a characterised primary human cell culture model to investigate cellular interactions with nano-porous alumina. This material, prepared by anodisation, is being developed as a coating on titanium alloy implants. The structure of the alumina, as determined by X-ray diffraction and transmission electron microscopy, was amorphous. When studying cell/material interactions we used both biochemical and morphological parameters. Cell viability, proliferation and phenotype were assessed by measurement of redox reactions in the cells, cellular DNA, tritiated thymidine ([3H]-TdR) incorporation and alkaline phosphatase (ALP) production. Results showed a normal osteoblastic growth pattern with increasing cell numbers during the first 2 weeks. A peak in cell proliferation was seen on day 3, after which cell growth decreased, followed by an increase in ALP production, thus indicating that the osteoblastic phenotype was retained on the alumina. Cell adhesion was observed, the osteoblast-like cells having a flattened morphology with filipodia attached to the pores of the material. SDS-PAGE and western blot measurements showed that the nano-porous alumina was able to adsorb fibronectin. Trace amounts of aluminium ions were measured in the surrounding medium, but no adverse effect on cell activity was observed.


The C3 component of complement; gene frequency variation in Iraq.
Heterogeneity of gene frequencies in the polymorphism of the third component of complement among local samples in Iraq is analysed. It appears to derive essentially from population and not from regional differences.


Association of Choroidal Interleukin-17-Producing T Lymphocytes and Macrophages with Geographic Atrophy.
To evaluate the presence of interleukin-17 (IL-17)-producing cells in patients with geographic atrophy (GA). In this short report, we analyzed IL-17, CD3, and IBA-1 expression by immunohistochemistry on paraffin-embedded sections from 13 donors with a known history of GA, confirmed by fundus appearance and histology, and 7 age-matched control donors. We showed that IL-17+ cells are found near areas of retinal pigmented epithelium atrophy in the eyes of GA patients. IL-17+ cells mainly localized to CD3+ cells, which identifies T lymphocytes, as well as IBA-1+ cells, which identifies mononuclear phagocytes. Therefore, IL-17 could be involved in the pathological mechanisms that contribute to the degeneration observed in GA.


Clinical predictors of alloimmunization after red blood cell transfusion.
Development of new red blood cell (RBC) alloantibodies (alloimmunization) is one of the most frequent adverse reactions after an RBC transfusion. Few studies have investigated clinical risk factors for alloimmunization. In this case-control study, the characteristics of all patients in whom alloimmunization occurred for the first time after an RBC transfusion in two hospitals between January 1, 2003, and May 5, 2005, were examined and compared to a randomly selected control group who received RBC transfusions in the same hospitals during the same period without alloimmunization. Odds ratios (ORs) for the association between these characteristics and alloimmunization were calculated and analyzed with a logistic regression model. Eighty-seven cases were found, and 101 controls were selected. Female sex (OR, 1.89; 95% confidence interval [CI], 1.05-3.38), diabetes mellitus (OR, 2.15; 95% CI, 0.91-5.05), solid malignancy (OR, 2.07; 95% CI, 1.00-4.30), and previous allogeneic hematopoietic peripheral blood progenitor cell (PBPC) transplantation (OR, 2.24; 95% CI, 0.64-7.81) were associated most strongly with alloimmunization, whereas lymphoproliferative disorders (OR, 0.33; 95% CI, 0.13-0.81) and symptomatic atherosclerosis (OR, 0.52; 95% CI, 0.25-1.08) were associated with the absence of alloimmunization. All of these associations except for female sex became stronger after adjustment for possible confounders. Female sex, diabetes mellitus, solid malignancy, and previous allogeneic PBPC transplantation seem to be risk factors for alloimmunization, whereas lymphoproliferative disorders and symptomatic atherosclerosis seem to protect against it. Further studies are needed to confirm these associations and investigate underlying mechanisms.


Vitamin D deficiency during pregnancy and lactation stimulates nephrogenesis in rat offspring.
There is increasing evidence of vitamin D insufficiency in women of child-bearing age and their infants. This study examined the effect of maternal vitamin D deficiency on nephron endowment in rat offspring (n=7 per group). Sprague-Dawley dams were fed either a vitamin D deplete diet or a vitamin replete (control) diet prior to pregnancy, during pregnancy and throughout lactation. At 4 weeks of age the offspring were weaned and maintained on their respective diets until they were killed at 7 weeks. In the fixed right kidney, kidney volume, renal corpuscle volume and nephron number were stereologically determined. There was no difference between groups in body weight, kidney weight or kidney volume. There was a significant 20% increase in nephron number in kidneys of vitamin D deplete offspring (vitamin D deficient, 29,000+/-1,858, control, 23,330+/-1,828; P=0.04). This was accompanied by a significant decrease in renal corpuscle size in the vitamin D deplete group compared with the controls (6.125+/-0.576 x 10(-4) mm(3) and 8.178+/-0.247 x 10(-4) mm(3), respectively; P=0.03). We concluded that maternal vitamin D deficiency in rats appears to stimulate nephrogenesis. Whether this confers a renal functional advantage or not is unknown.


Insights into the quality of recombinant proteins produced by two different Bombyx mori expression systems.
The silkworm, Bombyx mori, is an attractive host for recombinant protein production due to its high expression efficiency, quality, and quantity. Two expression systems have been widely used for recombinant protein production in B. mori: baculovirus/silkworm expression system and transgenic silkworm expression system. Both expression systems enable high protein production, but the qualities of the resulting recombinant proteins have not been well evaluated. In this study, we expressed bovine interferon γ (IFN-γ) using the two systems and examined the quality of the resulting proteins in terms of N-glycosylation and protein cleavage. Both expression systems successfully produced IFN-γ as an N-glycoprotein. Although the production in the baculovirus/silkworm expression system was much more efficient than that in the transgenic silkworm expression system, unexpected variants of IFN-γ were also produced in the former system due to the different N-glycosylation and C-terminal truncations. These results indicate that while high protein production could be achieved in the baculovirus/silkworm expression system, unintentional protein modification might occur, and therefore protein expression in the transgenic silkworm expression system is preferable from the point-of-view of N-glycosylation of the recombinant protein and evasion of unexpected attack by a protease in B. mori.


Does Earlier Cannulation With Veno-Venous Extracorporeal Membrane Oxygenation in Adult Patients With Acute Respiratory Distress Syndrome Decrease Duration of Artificial Mechanical Ventilation?
Acute respiratory distress syndrome (ARDS) is characterized by an acute, diffuse, inflammatory lung injury, leading to increased alveolar capillary permeability, increased lung weight, and loss of aerated lung tissue (Fan, Brodie, & Slutsky, 2018). Primary treatment for ARDS is artificial mechanical ventilation (AMV) (Wu, Huang, Wu, Wang, & Lin, 2016). Given recent advances in technology, the use of veno-venous extracorporeal membrane oxygenation (VV-ECMO) to treat severe ARDS is growing rapidly (Combes et al., 2014). This 49-month quantitative, retrospective inpatient EMR chart review compared if cannulation with VV-ECMO up to and including 48 hours of admission and diagnosis in adult patients 30 to 65 years of age diagnosed with ARDS, decreased duration on AMV, as compared to participants who were cannulated after 48 hours of admission and diagnosis with ARDS. A total of 110 participants were identified as receiving VV-ECMO during the study timeframe. Of the 58 participants who met all inclusion criteria, 39 participants were cannulated for VV-ECMO within 48 hours of admission and diagnosis with ARDS, and 19 participants were cannulated with VV-ECMO after 48 hours of admission and diagnosis with ARDS. Data collected identified no statistically significant (p < 0.579) difference in length of days on AMV between participant groups. Further studies are needed to determine if earlier initiation of VV-ECMO in adult patients with ARDS decrease time on AMV. Although the results related to length of time on AMV did not produce statistical significance, the decreased duration of AMV in the participants who were cannulated within 48 hours (21 days vs. 27 days) may support several benefits associated with this participant population including increased knowledge of healthcare providers, decreased lung injury, earlier discharge which decreases hospital and patient cost, ability for patients to communicate sooner, decreased risk of pulmonary infection, decreased length of stay, decreased cost, and improved patient and family satisfaction.


Differential susceptibilities of the prosthetic heme of hemoglobin-based red cell substitutes. Implications in the design of safer agents.
One approach to the development of an effective red cell substitute has been chemical modification of human hemoglobin to optimize oxygen transport and plasma half-life. Human hemoglobin A0 and two of these modified hemoglobins, one prepared from the cross-linking of the alpha-chains at lysine residue 99 by bis(3,5-dibromosalicyl)fumarate (Hb-DBBF) and the other by acylation of lysine residue 82 of the beta-chain by mono-(3,5-dibromosalicyl)fumarate (Hb-FMDA), were tested by HPLC for their susceptibility to oxidative damage caused by H2O2. Such oxidative insult may occur during ischemia and reperfusion of tissues after transfusion of red cell substitutes to patients with hypovolemic shock and trauma. Hb-DBBF was extremely susceptible to damage of its heme and protein moieties with stoichiometric amounts of H2O2, whereas Hb-FMDA was highly resistant, even at 10-fold molar excess and at an acidic pH of 4.7. Hemoglobin A0 was of intermediate susceptibility, exhibiting alteration of heme and protein moieties at acidic but not neutral pH. Since the degradation of heme can release the potentially toxic agent iron, Hb-FMDA may be a more promising candidate than Hb-DBBF for development as a red cell substitute. A similar approach may be used to assess the susceptibility of other hemoglobin-based red cell substitutes to oxidative damage in order to determine the molecular basis of heme and protein alteration.


Pyoverdine-mediated iron transport. Fate of iron and ligand in Pseudomonas aeruginosa.
Incubated in the presence of [55Fe]ferri[14C]pyoverdine, iron-poor Pseudomonas aeruginosa accumulated more 55Fe than 14C over a 60-min period. Distribution studies showed (a) more 14C than 55Fe in the soluble fraction during the first 20 min, (b) approximately 60% of the 55Fe associated with the membranes at 60 min, and (c) approximately 85% of the 14C in the soluble fraction at 60 min. Cells osmotically shocked after incubating with [55Fe]ferri[14C]pyoverdine for 60 min released 55Fe but not 14C, suggesting separation of metal and ligand in the periplasmic space. Whereas the mechanism of dissociation of iron and ligand is not known, the decrease in transport observed in the presence of dipyridyl suggests involvement of reduction in this process. Transport of iron was energized by the proton motive force instead of by intracellular levels of ATP. The hydrogen ion gradient was the major driving force of transport. Cyanide-poisoned cells accumulated more 14C than 55Fe over 60 min. Here, iron accumulated in the soluble fraction instead of on the membranes.


Important role of apoptosis signal-regulating kinase 1 in ischemic acute kidney injury.
We investigated the role of apoptosis signal-regulating kinase 1 (ASK1) in ischemia/reperfusion (I/R)-induced acute kidney injury (AKI). Blood urea nitrogen (BUN) and serum creatinine were significantly higher in ASK1+/+ mice than in ASK1-/- mice after I/R injury. Renal histology of ASK1+/+ mice showed significantly greater tubular necrosis and degradation. In ASK1-/- mice, phosphorylation of ASK1, JNK, and p38K, and the number of TUNEL-positive cells and infiltrated leukocytes decreased after I/R injury. Apoptotic changes were significantly decreased in cultured renal tubular epithelial cells (TECs) from ASK1-/- mice under hypoxic condition. Transfection with dominant-active ASK1 induced apoptosis in TECs. Protein expression of monocyte chemoattractant protein-1 (MCP-1) was significantly weaker in ASK1-/- mice after I/R injury. Transfection with dominant negative-ASK1 significantly decreased MCP-1 production in TECs. These results demonstrated that ASK1 is activated in I/R-induced AKI, and blockage of ASK1 attenuates renal tubular apoptosis, MCP-1 expression, and renal function.


Morphological and immunocytochemical characterisation of mixed glial cell cultures derived from neonatal canine brain.
Three different regions from neonatal dog brain were mechanically dissociated and cultured. At seven, 10, 14 and 19 days, cultures were harvested and studied with immunocytochemical techniques for the demonstration of astrocytes, oligodendrocytes and fibroblasts. The canine brain cultures were confluent at seven days and contained fibroblasts, both types of glial cells and several small fragments of undissociated brain tissue. Many cells were myelin associated glycoprotein (MAG) positive. In the older cultures increasing numbers of myelin basic protein (MBP) positive oligodendrocytes were also demonstrated. Double labelling studies demonstrated that MBP and MAG were produced by the same cells. MAG positive cells were therefore identified as oligodendrocytes. Most intensive oligodendroglial growth occurred in the vicinity of undissociated tissue fragments. The results were discussed in respect to glial differentiation and the question of whether the canine cultures may contain neurons.


Rear seat safer: seating position, restraint use and injuries in children in traffic crashes in Victoria, Australia.
Car crashes are a major cause of death and serious injury to children but most analyses of risk are based on US data. The Australian context is different in at least three ways: (1) the proportion of passenger-side airbags, a potential risk to children in front seats, is much lower; (2) unlike in the US, Australian airbags are designed to work with restrained passengers; (3) restraint use for children 0-12 years is high (>90%). Official data drawn from Victorian crash records (n=30,631) were used to calculate relative risks of death or serious injury for children (0-3 years, 4-7 years; 8-12 years) traveling in passenger cars during 1993-1998 and 1999-2004. Over 90% were reportedly wearing a restraint, and 20% were traveling in the front seat. For children under 4 years traveling in the front seat, the relative risk of death was twice as great as when traveling in the rear, and that of serious injury was 60% greater. The relative risk of death whilst traveling in the front seat was almost four times greater for children aged under 1 year. We suggest that serious consideration should be given to mandating rear seating for children, particularly those aged 4 and under.


Elemental analysis of samples of biological origin relative to their protein content by means of charged particle bombardment.
The particle-induced X-ray emission (PIXE) method and the 14N(d,p)15N nuclear reaction are combined for simultaneous trace element and nitrogen determination. Measurement of nitrogen content often allows the relating of the elemental concentrations determined by PIXE to the protein content of the sample. For the measurements only a small amount of sample material is needed; therefore, it is possible to keep track of the quantity of a certain element in the successive steps of a biomedical separation process. In about 10 min, trace element concentrations in the ppm range can be determined with a statistical accuracy of about 10%.


Psychosocial interventions for internalised stigma in people with a schizophrenia-spectrum diagnosis: A systematic narrative synthesis and meta-analysis.
It is acknowledged that people with a schizophrenia-spectrum diagnosis experience higher levels of stigma compared to any other mental health diagnosis. As a consequence, their experience of internalised stigma is likely to be the most detrimental and pervasive. Internalised stigma interventions have shown some benefits in those who experience serious mental illness including those with a schizophrenia-spectrum diagnosis. A systematic narrative review and meta-analysis were conducted examining the efficacy of internalised stigma interventions for people with a schizophrenia-spectrum diagnosis. Randomised Controlled Trials, controlled trials, and cohort studies were included and assessed against quality criteria. The search identified 12 studies; 7 randomised controlled trials, 3 cohort studies and 2 controlled trials. A variety of psychosocial interventions were utilised with the majority employing Cognitive Behaviour Therapy (CBT), psychoeducation and social skills training. The core outcomes used to examine the efficacy of the intervention were internalised stigma, self-esteem, empowerment, and functioning. The meta-analysis revealed an improvement in internalised stigma favouring the internalised stigma intervention but was not significant (5 RCTs, n=200). Self-efficacy and insight were significantly improved favouring the internalised stigma intervention. Internalised stigma interventions show promise in those with schizophrenia-spectrum diagnoses. Existing interventions have demonstrated small effects and employed small samples. Large scale RCTs are required to further develop the evidence base of more targeted interventions.


Expression of a Catalytically Inactive Mutant Form of Glutathione Peroxidase 4 (Gpx4) Confers a Dominant-negative Effect in Male Fertility.
The selenoenzyme Gpx4 is essential for early embryogenesis and cell viability for its unique function to prevent phospholipid oxidation. Recently, the cytosolic form of Gpx4 was identified as an upstream regulator of a novel form of non-apoptotic cell death, called ferroptosis, whereas the mitochondrial isoform of Gpx4 was previously shown to be crucial for male fertility. Here, we generated and analyzed mice with a targeted mutation of the active site selenocysteine of Gpx4 (Gpx4_U46S). Mice homozygous for Gpx4_U46S died at the same embryonic stage (E7.5) as Gpx4(-/-) embryos as expected. Surprisingly, male mice heterozygous for Gpx4_U46S presented subfertility. Subfertility was manifested in a reduced number of litters from heterozygous breeding and an impairment of spermatozoa to fertilize oocytes in vitro. Morphologically, sperm isolated from heterozygous Gpx4_U46S mice revealed many structural abnormalities particularly in the spermatozoa midpiece due to improper oxidation and polymerization of sperm capsular proteins and malformation of the mitochondrial capsule surrounding and stabilizing sperm mitochondria. These findings are reminiscent of sperm isolated from selenium-deprived rodents or from mice specifically lacking mitochondrial Gpx4. Due to a strongly facilitated incorporation of Ser in the polypeptide chain as compared with selenocysteine at the UGA codon, expression of the catalytically inactive Gpx4_U46S was found to be strongly increased. Because the stability of the mitochondrial capsule of mature spermatozoa depends on the moonlighting function of Gpx4 both as an enzyme oxidizing capsular protein thiols and as a structural protein, tightly controlled expression of functional Gpx4 emerges as a key for full male fertility.


A thirty year journey in the polyol pathway.
The development of the concept that aldose reductase (AR) is involved in diabetic complications is presented from its early beginning when Dr Van Heyningen first found polyols in sugar cataracts in 1959. The involvement of the polyol theory of sugar cataract is described. The prevention of sugar cataract formation by aldose reductase inhibitors dramatically demonstrates the role of this enzyme in sugar-induced cataracts. The possibility of AR involvement of other diabetic complications was an obvious extension of the polyol theory. The use of AR inhibitors in preventing diabetic changes in nerve and retina in animals strongly suggest that AR may play a role in the development of other diabetic complications.


Lipocalin-2-mediated upregulation of various antioxidants and growth factors protects bone marrow-derived mesenchymal stem cells against unfavorable microenvironments.
Despite many advantages of mesenchymal stem cells (MSCs) that make them suitable for cell therapy purposes, their therapeutic application has been limited due to their susceptibility to several stresses (e.g., nutrient-poor environment, oxidative stress, and hypoxic and masses of cytotoxic factors) to which they are exposed during their preparation and following transplantation. Hence, reinforcing MSCs against these stresses is a challenge for both basic and clinician scientists. Recently, much attention has been directed toward equipping MSCs with cytoprotective factors to strengthen them against unfavorable microenvironments. Here, we engineered MSCs with lipocalin 2 (Lcn2), a cytoprotective factor that is naturally induced following exposure of cells to stresses imposed by the microenvironment. Lcn2 overexpression not only did not interfere with the multidifferentiation capacity of the MSCs but also granted many protective properties to them. Lcn2 potentiated MSCs to withstand oxidative, hypoxia, and serum deprivation (SD) conditions via antagonizing their induced cytotoxicity and apoptosis. Adhesion rate of MSCs to coated culture plates was also enhanced by Lcn2 overexpression. In addition, Lcn2 induced antioxidants and upregulated some growth factors in MSCs. Our findings suggested a new strategy for prevention of graft cell death in MSC-based cell therapy.


High-throughput laboratory evolution reveals evolutionary constraints in Escherichia coli.
Understanding the constraints that shape the evolution of antibiotic resistance is critical for predicting and controlling drug resistance. Despite its importance, however, a systematic investigation of evolutionary constraints is lacking. Here, we perform a high-throughput laboratory evolution of Escherichia coli under the addition of 95 antibacterial chemicals and quantified the transcriptome, resistance, and genomic profiles for the evolved strains. Utilizing machine learning techniques, we analyze the phenotype-genotype data and identified low dimensional phenotypic states among the evolved strains. Further analysis reveals the underlying biological processes responsible for these distinct states, leading to the identification of trade-off relationships associated with drug resistance. We also report a decelerated evolution of β-lactam resistance, a phenomenon experienced by certain strains under various stresses resulting in higher acquired resistance to β-lactams compared to strains directly selected by β-lactams. These findings bridge the genotypic, gene expression, and drug resistance gap, while contributing to a better understanding of evolutionary constraints for antibiotic resistance.


Severe disability related to cerebral stroke: incidence and risk factors observed in a Japanese community, Hisayama.
To elucidate the incidence of severe disability due to cerebral stroke and its related factors, prospective data of 1,621 Hisayama residents aged 40 and over were examined. Severe disability resulting from stroke was defined as patients who were unable to dress, take care of their toilet needs, and feed themselves without assistance, or who required a wheel chair for ambulation three months after the most recent episode. During 20 years of follow-up 255 stroke patients were observed among the sample population. The annual incidence of stroke per thousand was 9.8, and rate of severe disability was 2.8 for men and 6.4 and 2.0 for women, respectively. Of the 74 cases with severe disability, approximately 92% were attributed to cerebral infarction. Related factors to severe disability due to cerebral infarction were recurrent attacks, hypertension, changes in ocular fundi and diabetes mellitus among predispositions and quadriplegia or muscular contraction, and intelligent or mental disorders among inhibiting factors for functional recovery. Furthermore, in 59 autopsy cases with multiple cerebral infarctions, the frequency of disability increased as the number of infarcts increased. Hypertension and diabetes mellitus, as risk factors for cerebral infarction and factors inhibiting post-ictal functional recovery were discussed.


Construction of an efficient expression vector for coupled transcription/translation in a wheat germ cell-free system.
Using the expression vector, pEU, which we have constructed, highly efficient in vitro protein synthesis can be achieved: The system works for 150 hours and without further template addition once the reaction has started, yielding 5 mg of enzymatically active protein in a 1 ml reaction.


Biomechanics of traumatic brain injury: influences of the morphologic heterogeneities of the cerebral cortex.
Traumatic brain injury (TBI) can be caused by accidents and often leads to permanent health issues or even death. Brain injury criteria are used for assessing the probability of TBI, if a certain mechanical load is applied. The currently used injury criteria in the automotive industry are based on global head kinematics. New methods, based on finite element modeling, use brain injury criteria at lower scale levels, e.g., tissue-based injury criteria. However, most current computational head models lack the anatomical details of the cerebrum. To investigate the influence of the morphologic heterogeneities of the cerebral cortex, a numerical model of a representative part of the cerebral cortex with a detailed geometry has been developed. Several different geometries containing gyri and sulci have been developed for this model. Also, a homogeneous geometry has been made to analyze the relative importance of the heterogeneities. The loading conditions are based on a computational head model simulation. The results of this model indicate that the heterogeneities have an influence on the equivalent stress. The maximum equivalent stress in the heterogeneous models is increased by a factor of about 1.3-1.9 with respect to the homogeneous model, whereas the mean equivalent stress is increased by at most 10%. This implies that tissue-based injury criteria may not be accurately applied to most computational head models used nowadays, which do not account for sulci and gyri.


Inhibition of the activity of pro-inflammatory secretory phospholipase A(2) by acute phase proteins.
Pro-Inflammatory non-pancreatic phospholipase A(2) (sPLA(2)) is markedly over-expressed in acute systemic and chronic local inflammatory processes. Since in acute phase reaction sPLA(2) is often over-expressed simultaneously with acute phase proteins (APP), it is important to determine whether APP interacts with sPLA(2). We tested ten APPs for interaction with sPLA(2) using as a substrate multilamellar Hposomes composed either of PC:Lyso PC or PE:Lyso PE. Using PC:Lyso PC substrate, CRP, lactoferrin and SAP were found to inhibit sPLA(2) activity with an IC(50) of 25 mug/ml, 7.5 mug/ml and 50 mug/ml, respectively, corresponding to 0.21 muM, 0.1 muM and 0.21 muM respectively. Using PE:Lyso PE substrate only SAP was inhibitory, with an IC(50) of 10 mug/ml (0.04 muM). Phosphorylcholine abolished the inhibitory activity of CRP but not of SAP or lactoferrin. Addition of phosphorylethanolamine or of excess calcium had no effect on the inhibitory activity of APP. Limulin, lysozyme, transferrin, beta(2)-microglobulin, alpha(2)-macroglobulin, human and bovine albumins had no effect on sPLA2 activity. Therefore neither the structure of pentraxins, or ironbinding, bacteriostatic property or amyloidogenic property preclude whether APP modulates sPLA(2) activity. Inhibition of pro-inflammatory sPLA(2) by APP may be one of the protective mechanisms of the acute phase reaction.


Efficient production of deoxynucleoside-5'-monophosphates using deoxynucleoside kinase coupled with a GTP-regeneration system.
Deoxynucleoside-5'-monophosphates (5'-dNMPs) are the basic components of DNA and are widely used in medicine and as chemical and biochemical reagents. A large amount of effort has been expended to obtain 5'-dNMPs of high quality and at a low cost. However, these procedures are inefficient and inconvenient. In this study, deoxyadenosine-5'-monophosphate (5'-dAMP), 2,6-diaminopurine deoxynucleoside-5'-monophosphate (5'-dDAMP), and deoxycytidine-5'-monophosphate (5'-dCMP) were biosynthesized using recombinant N-deoxyribosyltransferase II (NDT-II), deoxycytidine kinase, and acetate kinase in a one-pot reaction system. The ndt-II gene from Lactobacillus delbrueckii, dck from Bacillus subtilus, and ack from Escherichia coli K12 were overexpressed in E. coli BL21 (DE3). Thymidine was used as the deoxyribose donor; GTP was used as the phosphate donor, and acetyl phosphate was used to regenerate GTP. Under optimized conditions, each 10 mM adenine, 10 mM 2,6-diaminopurine, or 10 mM cytosine were converted into 9.01 mM 5'-dAMP, 8.68 mM 5'-dDAMP, or 6.23 mM 5'-dCMP, respectively. The high yield indicated that this process of biosynthesis of 5'-dAMP, 5'-dDAMP, or 5'-dCMP was efficient and economical, and this one-pot system may also potentially be used for the preparation of other types of 5'-dNMPs.


Liquid-liquid phase separation in hemoglobins: distinct aggregation mechanisms of the beta6 mutants.
Reversible liquid-liquid (L-L) phase separation in the form of high concentration hemoglobin (Hb) solution droplets is favored in an equilibrium with a low-concentration Hb solution when induced by inositol-hexaphosphate in the presence of polyethylene glycol 4000 at pH 6.35 HEPES (50 mM). The L-L phase separation of Hb serves as a model to elucidate intermolecular interactions that may give rise to accelerated nucleation kinetics of liganded HbC (beta6 Lys) compared to HbS (beta6 Val) and HbA (beta6 Glu). Under conditions of low pH (pH 6.35) in the presence of inositol-hexaphosphate, COHb assumes an altered R-state. The phase lines for the three Hb variants in concentration and temperature coordinates indicate that liganded HbC exhibits a stronger net intermolecular attraction with a longer range than liganded HbS and HbA. Over time, L-L phase separation gives rise to amorphous aggregation and subsequent formation of crystals of different kinetics and habits, unique to the individual Hb. The composite of R- and T-like solution aggregation behavior indicates that this is a conformationally driven event. These results indicate that specific contact sites, thermodynamics, and kinetics all play a role in L-L phase separation and differ for the beta6 mutant hemoglobins compared to HbA. In addition, the dense liquid droplet interface or aggregate interface noticeably participates in crystal nucleation.


Evaluation of the In-Use Stability of Monoclonal Antibody IV Admixtures Prepared from Drug Products Containing Polysorbate 20 Degraded by Host-Cell Lipases.
Host-cell lipases can be present in monoclonal antibody drug products and can degrade polysorbates present in the formulations as stabilizers. We hypothesized that the in-use stability of the IV admixture prepared from such a drug product might be impacted by decreasing levels of polysorbate 20. Host-cell lipase activity has, in fact, been observed during development of one of our therapeutic monoclonal antibody drug products. Throughout the course of the product shelf life, polysorbate 20 levels decreased but no other quality attributes of the drug product were impacted. An experimental approach was developed to simulate how the prepared IV admixture in-use stability is affected as polysorbate 20 concentration in the drug product decreased over the shelf life, and from that a minimum level of polysorbate 20 required in the drug product was determined to estimate the in-use stability of the IV admixture as the polysorbate 20 in the drug product degrades. The results indicate that although the observed degradation of polysorbate 20 does not affect quality attributes of this drug product, in-use stability of the IV admixture as a function of polysorbate degradation can be impacted and should be assessed to ensure sufficient quality.


The Effects of Race and Socioeconomics on the Relationship Between Gestational Diabetes Mellitus and Birth Outcomes: An Analysis of Southern US PRAMS data.
Gestational diabetes mellitus (GDM) is one of the most common pregnancy complications. Uncontrolled GDM increases the risk of negative pregnancy outcomes for both the pregnant individual and the infant. African Americans (AAs) have higher maternal morbidity and infant mortality rates than non-Hispanic whites (NHWs). The goal of the current study was to examine racial differences in the effect of GDM on birth outcomes. The data from the Pregnancy Risk Assessment Monitoring System (PRAMS) was analyzed with a focus on four states in the southern U.S. (Alabama, Georgia, Louisiana, and Mississippi). The results presented suggest that AAs are at lower risk of GDM than NHW individuals. Even with the lower risk of GDM, AAs are at higher risk of pre-term births. In addition, socioeconomic factors and access to prenatal care play a role in birth outcomes including moderating the effect of GDM on outcomes. A discussion of potential policy interventions that may improve pregnancy outcomes is discussed including increased use of doulas to support pregnant people.


Management of upper airway obstruction.
The tongue is the most common cause of upper airway obstruction, a situation seen most often in patients who are comatose or who have suffered cardiopulmonary arrest. Other common causes of upper airway obstruction include edema of the oropharynx and larynx, trauma, foreign body, and infection. The management of the patient with upper airway obstruction depends upon the cause of the obstruction, the training and skills of the rescuer, and the availability of adjuncts necessary to perform advanced airway techniques. In most cases, merely positioning the patient or performing one of the three maneuvers designed to elevate the tongue will open the airway of the comatose patient or the victim of cardiac arrest. In patients with suspected foreign body obstruction, abdominal or chest compression should be performed immediately, with digital extraction of the foreign body reserved for those in whom these maneuvers are unsuccessful. Most patients with obstruction secondary to edema, trauma, or infection can be managed initially with orotracheal or nasotracheal intubation. Intubation should be attempted prior to surgical management of the airway in most cases of upper airway obstruction. Occasionally, however, cricothyroidotomy or tracheostomy is necessary to establish an airway. The choice of technique depends primarily on the experience and skills of the rescuing physician or paramedic. In most cases, cricothyroidotomy is technically more simple and more easily performed than tracheotomy, especially for the physician who has not been trained in surgery or otolaryngology and for the nonphysician rescuer. No matter what the method employed in establishing an airway in a patient with upper airway obstruction, it must be performed quickly and a source of ventilation provided for the patient once the airway has been secured.


[Association between interferon regulatory factor 6 gene polymorphism and non-syndromic oral clefting].
To investigate the association between interferon regulatory factor 6 (IRF6) gene polymorphism and non-syndromic oral clefting (NSOC). Experimental group consisted of 186 Ningxia NSOC patients, their parents (183 fathers and 174 mothers), 172 core families (patient+parents), and control group consisted of 200 normal children. DNA was extracted and PCR-restriction fragment length polymorphism (PCR-RFLP) was used to identify the genotypes of the samples, case-control analyses and transmission-disequilibrium test (TDT) were carried out. Compared with control group, there were significant differences in both rs642961's and rs4844880's AA genotype and A allele among NSOC patients (P < 0.05), but no difference in cleft palate (P = 0.15, P = 0.967, respectively). In TDT analysis, the A allele of rs642961 had a strong over-transmission in NSOC (P < 0.05), so did the rs4844880'A allele (P < 0.05), but neither of them had significant difference in cleft palate (P = 0.91, P = 0.95, respectively). IRF6 gene polymorphism is associated with NSOC.


Insulin release in response to calcium in the diagnosis of insulinoma.
A calcium infusion (4 mg Ca++/kg/hr) significantly increased plasma insulin levels and reduced blood glucose in 4 patients with insulin-secreting pancreatic islet cell tumors. These parameters were not altered by a similar infusion of calcium in normal volunteers, 2 patients with alimentary hypoglycemia, and 2 with functional hypoglycemia. No difference in response was observed between patients with benign and malignant beta-cell tumors. Infusion of diazoxide (600 mg) with calcium blocked the stimulation of the latter on insulin secretion. The results indicate the usefulness of calcium infusion in the diagnosis of insulin-secreting tumor.


Effectiveness of the DHMAI monomer in the development of an antibacterial dental composite.
Development of antibacterial dental composites is the ultimate goal to decrease carious disease occurrence and increase the restoration longevity. For this purpose, the quaternary ammonium dimethyl-hexadecyl-methacryloxyethyl-ammonium iodide (DHMAI) and the methacryloyloxyethylphosphorylcholine (MPC) have been incorporated in experimental methacrylate-based composite resins. This aims to first investigate the effect of each alone and then their combined effect. Synthesized DHMAI and commercial MPC were added either alone or combined at different concentrations to experimental dental composite. Flexural strength (FS) and modulus (FM) were tested to select the optimal concentrations. Only selected composites were evaluated for Vickers hardness (HV) and the degree of conversion (DC) using fourier transform infrared spectroscopy analysis (FTIR-ATR). Antibacterial activity was assessed using tests on colony-forming unit (CFU), scanning electron microscopy (SEM) and Alamarblue assay to measure the metabolic activity. Streptococcus mutans biofilm was chosen to be grown on the composite surfaces during 96h at 37°C. Incorporation of 7.5% DHMAI in composite improved the degree of conversion and gave a strong antibacterial effect with a reduction of (∼98%) in CFU and (∼50%) of metabolic activity with acceptable mechanical properties. Addition of MPC to DHMAI affects mechanical properties of composites without providing a better antibacterial activity. Composites with DHMAI greatly reduced S. mutans biofilm and improved the degree of conversion without scarifying the composites' mechanical properties. DHMAI may have wide applicability to other dental materials in order to inhibit caries and improve the longevity of restorations.


Bladder stones after bladder augmentation are not what they seem.
Bladder and renal calculi after bladder augmentation are thought to be primarily infectious, yet few studies have reported stone composition. The primary aim was to assess bladder stone composition after augmentation, and renal stone composition in those with subsequent nephrolithiasis. The exploratory secondary aim was to screen for possible risk factors for developing infectious stones. Patients treated for bladder stones after bladder augmentation at the present institution between 1981 and 2012 were retrospectively reviewed. Data were collected on demographics, surgeries and stone composition. Patients without stone analysis were excluded. Stones containing struvite, carbonate apatite or ammonium acid ureate were classified as infectious. The following variables were analyzed for a possible association with infectious bladder stone composition: gender, history of cloacal exstrophy, ambulatory status, nephrolithiasis, recurrent urea-splitting urinary tract infections, first vs recurrent stones, timing of presentation with a calculus, history of bladder neck procedures, catheterizable channel and vesicoureteral reflux. Fisher's exact test was used for analysis. Of the 107 patients with bladder stones after bladder augmentation, 85 met inclusion criteria. Median age at augmentation was 8.0 years (follow-up 10.8 years). Forty-four patients (51.8%) recurred (14 multiple recurrences, 143 bladder stones). Renal calculi developed in 19 (22.4%) patients with a bladder stone, and 10 (52.6%) recurred (30 renal stones). Overall, 30.8% of bladder stones were non-infectious (Table). Among patients recurring after an infectious bladder stone, 30.4% recurred with a non-infectious one. Among patients recurring after a non-infectious stone, 84.6% recurred with a non-infectious one (P = 0.005). Compared with bladder stones, renal stones were more likely to be non-infectious (60.0%, P = 0.003). Of patients with recurrent renal calculi after an infectious stone, 40.0% recurred with a non-infectious one. No clinical variables were significantly associated with infectious stone composition on univariate (≥0.28) or bivariate analysis (≥0.36). This study had several limitations: it was not possible to accurately assess adherence with bladder irrigations, and routine metabolic evaluations were not performed. The findings may not apply to patients in all clinical settings. While stone analysis was available for 3/4 of the stones, similar rates of incomplete stone analyses have been reported in other series. In patients with bladder augmentation, 1/3 of bladder stones and >1/2 of renal stones were non-infectious. Furthermore, an infectious stone does not imply an infectious recurrent stone and no known clinical variables appear to be associated with stone composition, suggesting that there is a possible metabolic component in stone formation after bladder augmentation.


Effects of pirenzepine on plasma insulin, glucagon and pancreatic polypeptide levels in normal man.
The secretion of various pancreatic hormones (insulin, glucagon and pancreatic polypeptide) is affected to a different extent by the cholinergic system. In 7 healthy subjects the effects of treatment for 1 week with pirenzepine, an anticholinergic drug selective for muscarinic receptors, on basal secretion of these hormones and on that induced by i.v. glucose (IVGTT) and arginine were evaluated. The drug did not reduce basal levels of insulin and glucagon whereas it caused an appreciable reduction in basal pancreatic polypeptide (PP). The responses of insulin and blood glucose to IVGTT and to arginine were not changed by treatment, nor was that of plasma glucagon to arginine. The infusion of arginine did induce an increase in PP level, which reached a statistically significant maximum at 90 min. This response was not particularly different after administration of pirenzepine. Thus, the results confirm the finding that arginine stimulates PP secretion in vivo and that pirenzepine reduces the basal level of the hormone, whereas it did not appear to affect the response to arginine. The findings exclude any direct action of the drug on insulin or glucagon secretion or on glucose metabolism in general.


Toxic and active material basis of Aconitum sinomontanum Nakai based on biological activity guidance and UPLC-Q/TOF-MS technology.
As a folk medicine, Aconitum sinomontanum Nakai (Ranunculaceae, Gaowutou, in Chinese) is used by traditional healers to treat many disorders, including pain and inflammatory diseases, but it exhibits the toxic side effects. This study aimed to obtain toxic extract parts from A. sinomontanum roots and to further evaluate the antinociceptive and anti-inflammatory effects of toxic extract parts on mice. This work also aimed to identify various chemical compositions of the toxic and active extract parts and evaluate the safety profile of this plant. Experimental drugs (petroleum ether, chloroform, ethyl acetate, n-butanol, alcohol and water extracts) were obtained through systematic solvent extraction from 95 % ethanol extract from A. sinomontanum roots. An acute toxicity test was conducted to compare the toxicity of different extracts administered at the maximum dose to screen a highly toxic extract. In pharmacodynamic activity analysis, the antinociceptive activity of the A. sinomontanum toxic extract was assessed using an acetic acid-induced abdominal writhing model and a hot plate test. Anti-inflammatory activity was assessed in terms of xylene-induced inflammation. Ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was performed to establish a chromatographic fingerprint and to identify various chemical components of the toxic and active extract. Chloroform, water and n-butanol extracts elicited significant toxic effects and had LD50 of 89.65, 1805.40 and 24409.41 mg/kg, respectively. Antinociceptive and anti-inflammatory activities indicated that the chloroform extract significantly alleviated (p < 0.01) the pain induced by acetic acid with an inhibition rate of 44.7 % (5.9 mg/kg) and 50.4 % (17.7 mg/kg). The chloroform extract also significantly (p < 0.01) increased the latency time during the hot plate test. The latency time at 5.9 and 17.7 mg/kg increased from 15.6 ± 4.1 s to 47.3 ± 6.4 s and from 16.3 ± 3.8 s to 49.8 ± 7.6 s (p < 0.01), respectively, 2 h after treatment. In the inflammatory test, the chloroform extract significantly reduced (p < 0.01) the xylene-induced mouse ear oedema with an inhibition rate of 45.48 % (5.9 mg/kg) and 51.46 % (17.7 mg/kg), respectively. This result indicated that A. sinomontanum chloroform extract was also the active extract part of A. sinomontanum. Phytochemical analysis revealed the presence of alkaloids in the chloroform extract. A total of 30 compounds were detected, and 23 compounds, including lappaconine, ranaconidine, 8-O-acetylexcelsine, sinomontanine H, finaconitine, lappacontine, N-dacetyllappaconitine, ranaconitine and isolappaconitine, were identified. A. sinomontanum chloroform extract possesses antinociceptive and anti-inflammatory activities and exhibits significant toxic effects. Phytochemical analysis indicated that some alkaloids may be the main bioactive ingredient responsible for the toxicity and efficacy of A. sinomontanum. This work contributes to the determination of the safety of the medicinal use of A. sinomontanum roots.


Haptoglobin 2-2 Phenotype Is Associated With Increased Acute Kidney Injury After Elective Cardiac Surgery in Patients With Diabetes Mellitus.
Recent studies reported an association between the 2-2 phenotype of haptoglobin (Hp 2-2) and increased cardiorenal morbidity in nonsurgical diabetic patients. Our goal was to determine whether the Hp 2-2 phenotype was associated with acute kidney injury (AKI) after elective cardiac surgery in patients with diabetes mellitus. We prospectively enrolled 99 diabetic patients requiring elective cardiac surgery with cardiopulmonary bypass. Haptoglobin phenotypes were determined by gel electrophoresis. Cell-free hemoglobin, haptoglobin, and total serum bilirubin were quantified as hemolysis markers. The primary outcome was postoperative AKI, as defined by the Acute Kidney Injury Network classification. The incidence of AKI was significantly higher in Hp 2-2 patients compared with patients without this phenotype (non-Hp-2-2; 55.6% versus 27%, P<0.01). The need for renal replacement therapy was also significantly higher in the Hp 2-2 group (5 patients versus 1 patient, P=0.02). Thirty-day mortality (3 versus 0 patients, P=0.04) and 1-year mortality (5 versus 0 patients, P<0.01) were also significantly higher in patients with the Hp 2-2 phenotype. In multivariable analysis, Hp 2-2 was an independent predictor of postoperative AKI (P=0.01; odds ratio: 4.17; 95% confidence interval, 1.35-12.48). Hp 2-2 phenotype is an independent predictor of postoperative AKI and is associated with decreased short and long-term survival after cardiac surgery in patients with diabetes mellitus.


Tissue-specific expression pattern of human endothelial lipase in transgenic mice.
Endothelial lipase (EL), a new member of the triacylglycerol lipase gene family, is a key enzyme in HDL metabolism. The EL expression pattern in humans was reported to be unique and complementary to that documented for lipoprotein lipase. The regulatory elements responsible for the tissue-specific EL expression are not identified yet. In order to confine these sequences to a defined region of the EL promoter, we analyzed EL mRNA expression in human EL transgenic mice expressing EL under the control of the endogenous human promoter. We identified small intestine, mammary gland, adipose tissue and the adrenal gland as previously unknown tissues to express EL. Our data demonstrate that regulatory elements within 11.4 kb of 5' and 9.9 kb of 3' human EL flanking region promote the expression of EL in small intestine, ovary, testis, mammary gland, brain, lung, aorta, adipose tissue and the adrenals, whereas regulatory sequences located between 27.4 and 11.4 kb of 5' or 9.9 and 48.7 kb of 3' human EL flanking region seem to be responsible for kidney-specific EL expression.


Association between true non-contrast and virtual non-contrast vertebral bone CT attenuation values determined using dual-layer spectral detector CT.
To investigate the association of vertebral CT attenuation between virtual non-contrast (VNC) and true non-contrast (TNC) images and to evaluate if VNC vertebral CT attenuation could be used for phantom-less osteoporosis detection in dual-layer spectral-detector CT (SDCT). 200 patients with non-contrast and portal-venous phase SDCT were retrospectively assigned to a test and a validation group of 100 patients each. CT attenuation of L1 vertebrae were measured on VNC and TNC. The test group was used to determine the difference between VNC and TNC CT attenuation and to calculate a statistical model for TNC CT attenuation prediction. The validation group was used to assess the capability of the model to predict TNC from VNC CT attenuation and its accuracy to identify osteoporosis. Osteoporosis was defined as a TNC CT attenuation of ≤110HU. In both groups, CT attenuation was lower in VNC than in TNC (P < 0.001). VNC and TNC CT attenuation was correlated strongly (r = 0.958). Using the regression equation established in the test group (TNC = 23.677 + 1.540 × VNC), the predicted TNC CT attenuation did not differ from the real TNC CT attenuation in the validation group (P = 0.359). A VNC CT attenuation cut-off of 52HU yielded an AUC of 0.978 for osteoporosis detection. L1 CT attenuation is systematically underestimated in VNC compared with TNC images. However, TNC L1 CT attenuation can be predicted reliably from VNC. VNC may perform well in phantom-less osteoporosis detection.


Polysaccharides from Lachnum sp. Inhibited colitis-associated colon tumorigenesis in mice by modulating fecal microbiota and metabolites.
It is still uncertain whether the consumption of Lachnum sp. polysaccharides (LEP) alleviates colorectal cancer (CRC) through the gut microbiota. In this study, our efforts are focused on the influence of LEP on CRC, intestinal barrier and inflammation, and fecal microbiota and the metabolites, in azoxymethane (AOM)/dextran sodium sulfate (DSS)-induced CRC mice. Results showed that LEP inhibited CRC mouse colon shortening and weight loss, decreased tumor incidence, restored intestinal barrier integrity, and reduced excessive inflammation. LEP consumption significantly altered microbiota overall structure and community, with reduced pernicious bacteria (such as Parabacteroides, Escherichia_Shigella, Desulfovibrio and Helicobacter), and increased beneficial bacterium (such as Alistipes, Alloprevotella and Ruminiclostridium). Fecal-metabolome profile indicated that a total of 43 metabolites were clearly changed, with 10 down-regulated and 33 up-regulated metabolites. In addition, short-chain fatty acids (SCFAs), including acetic acid, propionic acid and n-butyric acid, were significantly increased after LEP administration. Moreover, a strong correlation between the fluctuant gut microbiota and metabolites was found. These findings provided not only deeper insights into the responsibility of LEP for CRC alleviation, and but also the potential of LEP as a promising candidate for CRC prevention and treatment.


5-Hydroxytryptamine3 receptor modulation of excitatory amino acid release in the rat nucleus tractus solitarius.
In vivo microdialysis was employed to measure release of endogenous L-glutamate (GLU) and L-aspartate (ASP) in the medial nucleus tractus solitarius of urethane anaesthetised rats. Basal extracellular levels of these amino acids were stable following a 90 min equilibration period (6.3 +/- 0.24 and 3.4 +/- 0.6 pmol/20 microliters sample of GLU and ASP, respectively). Basal levels of endogenous extracellular GLU and ASP were increased over 2-fold and 3-fold, respectively, following local administration of the selective 5-hydroxytryptamine (5-HT3) receptor agonist phenylbiguanide (300 microM). Intracerebral administration of the selective 5-HT3 receptor antagonist ondansetron (30 microM) blocked the effect of phenylbiguanide on GLU release whilst the effect on ASP was variable and complex. These data suggest that 5-HT3 receptor activation in the rat nucleus tractus solitarius can affect excitatory amino acid neurotransmission in this region of the medulla oblongata.


Potential mortality effects of off-highway vehicles on the flat-tailed horned lizard (Phrynosoma mcallii): a manipulative experiment.
Southern California desert public lands receive especially high levels of off-highway recreation due to large population centers nearby and popular riding environments such as sand dunes. Controversy has developed over the flat-tailed horned lizard (Phrynosoma mcallii), previously a candidate for listing under the Endangered Species Act. Some evidence suggests lower lizard abundance in areas of higher recreational use than in areas with low or no use. We designed a manipulative experiment to ensure maximum inference in evaluating the direct impact of recreational riding of off-highway vehicles on lizards. Thirty-six lizards, in situ, were treated with an off-highway vehicle treatment during hibernation season in three treatment groups: high impact, low impact, and control. Treatments consisted of timed riding by off-highway vehicles. In all treatment groups survival was 100%, despite hibernation of lizards at very shallow depths. Consequently, indirect effects of off-highway vehicles deserve increased attention. The relative importance of direct versus indirect (i.e., degradation of lizard habitat) impacts caused by off-highway vehicles remains unknown. These indirect effects may include the altering of vegetation, substrate, and prey. We recommend that a manipulative approach be adopted to investigate these possibilities.


Bilateral simultaneous sudden sensorineural hearing loss.
This study adopted an inner ear test battery and MR imaging in patients with bilateral sudden sensorineural hearing loss (SSNHL) to investigate their causes, disease extent, and evaluate hearing outcome. From 1995 to 2014, 16 patients with bilateral SSNHL received audiometry, caloric test and MR imaging. Vestibular-evoked myogenic potential (VEMP) test was added to the test battery after 2000. Percentages of abnormal mean hearing level (MHL), cervical VEMP test, ocular VEMP test, and caloric test in patients with bilateral SSNHL were 100% (32/32), 100% (12/12), 100% (4/4), and 81% (26/32), respectively, implying that not only the cochlear part but also the vestibular part was severely affected in both ears. Causes of bilateral SSNHL were neoplasm in 5 patients, stroke in 5, meningitis in 1, and unknown in 5. Post-treatment MHL did not significantly differ from pre-treatment MHL indicating poor hearing outcome. Seven patients (44%) had passed away within 5years after onset, 2 patients were lost, and 7 patients survived. Via MR imaging, causes of bilateral SSNHL were identified for 69% of cases. Both cochlear and vestibular endorgans/afferents were identified to be severely affected bilaterally by the vestibular test battery and resulted in poor hearing outcome. A high mortality rate (44%) indicates that bilateral SSNHL is an ominous sign for a more sinister underlying disease.


[The cytopathic and cytogenetic sequelae of chronic inhalational exposure to formaldehyde on female germ cells and bone marrow cells in rats].
A study was made of cytopathological and cytogenetic effects of formaldehyde chronic inhalation, in doses 0.5 and 1.5 mg/m3, on the female rat's germ and marrow cells. The harmful effect of formaldehyde on germ cells (according to the patterns of early embryogenesis) is noted under the dose 1.5 mg/m3 only, while the reliable clastogenic and cytogenetic effects on the marrow cells were revealed even in the dose 0.5 mg/m3. It is concluded that the differences between effects of small doses of formaldehyde on different cell systems were evidently caused by the specific cell dynamics of these systems.


Serum glutathione S-transferase P1 1 in prediction of cardiac function.
Glutathione S-transferase P1 1 (GSTP1) belongs to the multigene isozyme family involved in cellular response to oxidative stress and apoptosis. Our initial retrospective proteomic analysis suggested that GSTP1 is associated with heart failure (HF). Although pro-B-type natriuretic peptide (proBNP) serves currently as a surrogate diagnostic and prognostic parameter in HF patients, its specificity remains uncertain. We hypothesized that GSTP1 might be a useful serum marker in the monitoring of HF patients. Serum GSTP1 and proBNP were prospectively measured in 193 patients subdivided based on their ejection fraction (EF) either in equal-sized quintiles or predefined EF groups >52%, 43%-52%, 33%-42%, 23%-32% and ≤22%. At a cutoff of ≥231 ng/mL, GSTP1 identified HF patients with EF ≤22% with 81% sensitivity and 83% specificity, and at a cutoff of ≥655 pg/mL, proBNP identified the same patient group with 84% sensitivity and 22% specificity. GSTP1 at a ≥126 ng/mL cutoff identified EF ≤42% with 90% sensitivity and 95% specificity, or proBNP at a ≥396 pg/mL cutoff had 97% sensitivity and 20% specificity. In regression analyses, GSTP1, but not proBNP, discriminated between EF ≤42% and EF >42% in HF patients. These results suggest that GSTP1 is strongly associated with HF and could serve as a sensitive and specific marker to predict the ventricular function in HF patients.


Patient involvement in enhanced recovery.
There are proven benefits of encouraging patients to play a more active role and take responsibility for their recuperation after all surgical procedures. This article explores the concept of enhanced recovery and the development of tools to support patients and health professionals in using an enhanced recovery pathway.


[The anti-tumor efficacy of human recombinant interleukin-2 (rIL-2) in vivo].
Our results suggest that the growth of tumors was significantly suppressed when the high-dose rIL-2 (2 x 10(4)U/day) was injected i.p. in mice bearing subcutaneously (s. c.) transplanted tumors. The inhibition effect on the growth of mouse hepatoma HAC was more potent than that on S180 group. It was found that rIL-2 increased significantly the survival of BALB/C mice bearing HAC tumor (P < 0.001). We have also shown that the adoptive transfer of unfractionated LAK cells plus rIL-2 was more effective than rIL-2 alone. The results also indicated that adherent LAK cells (A-LAK) in combination with rIL-2 had higher antitumor effects as compared to standard LAK cells plus rIL-2.


Ileal urinary reservoir in pediatric population: objective assessment of long-term sequelae with time-to-event analysis.
To evaluate the long-term outcomes of an ileal urinary reservoir in children. This was a longitudinal study of pediatric patients who had undergone total ileal substitution of the bladder. Continence status was assessed, and all patients were evaluated for kidney function and biochemical profile. Standardized growth charts were used to assess linear growth. To assess bone mineral density, dual-emission x-ray absorptiometry scanning was performed. Clavien's scale was used to report and grade the long-term complications and their timing. We used a simple quality of life questionnaire to assess the effect of the procedure on the quality of life of the growing child. A total of 17 patients were included; 3 with orthotopic and 14 with continent cutaneous reservoirs. After a mean follow-up of 87.3 months, all patients were voiding with clean intermittent catheterization, with a 94% final continence rate. Two patients (11.7%) had an estimated glomerular filtration rate of ≤ 45 mL/min/1.73 m(2) at the last follow-up examination. However, no clinically manifest metabolic acidosis was detected. No anemia or neurologic deficit was detected, with a low-normal serum level of vitamin B(12) in 2 patients (11.7%) and a low level in 1 patient (5.7%). One patient (5.7%) had chronic diarrhea. Low bone mineral density was found in 4 patients (22.8%), with 3 patients (17.1%) not exceeding the fifth percentile of height for age. High-grade complications (grade 3a-5) represented 64.5% of the complications, and the need for reintervention occurred late in the follow-up period. A high level of quality of life satisfaction was reported (88.5%). Ileal neobladder construction allows child to pass into adolescence dry with more confidence and self-esteem, with no external urine collection set. However, long-term follow-up is mandatory to maintain the positive outcome.


Correction to: West Highland White Terriers under primary veterinary care in the UK in 2016: demography, mortality and disorders.
[This corrects the article DOI: 10.1186/s40575-019-0075-2.].


Possibility of SARS-CoV-2 transmission from the breast milk of COVID-19 affected women patients to their infants: worries and strategies to counter it.
Not available.


Metabolism, excretion, and pharmacokinetics of [14c]-radiolabeled aleglitazar: a phase I, nonrandomized, open-label, single-center, single-dose study in healthy male volunteers.
Aleglitazar is a dual peroxisome proliferator-activated receptor (PPAR)-α/γ agonist with a balanced activity (similar half-maximal effective concentrations) toward PPAR-α and -γ that is in clinical development for the treatment of patients who have experienced an acute coronary syndrome and have type 2 diabetes mellitus. This study aimed to characterize the metabolic profile and the routes and rates of elimination of aleglitazar and its major metabolites in humans. In this Phase I, nonrandomized, open-label, single-center, single-dose study, 6 healthy male subjects each received a single oral dose of 300 μg [(14)C]-labeled aleglitazar. Total urine and feces were collected for up to 15 days. Venous blood samples were collected to determine the plasma concentrations of aleglitazar and its metabolites and for radioactivity counting. The median age (range) and mean (SD) body mass index of subjects were 48 (41-60) years and 24.8 (3.0) kg/m(2), respectively. Recovery of total radioactivity, as a percentage of the dose administered, was high (93 [3]%). Aleglitazar was predominantly eliminated in feces (mean, 66% [range, 55%-74%]), with only 28% (range, 22%-36%) of the radioactivity recovered in urine. Only a mean (SD) of 1.8 (0.8)% of aleglitazar was eliminated unchanged as parent compound in feces and only 0.3 (0.4)% was eliminated in urine. Almost all excreted drug-related material could be attributed to its 2 main metabolites, M1 (21%) and M6 (38%). Treatment with aleglitazar was well tolerated, and no serious adverse events were reported. In healthy volunteers, aleglitazar was excreted mainly in the form of inactive metabolites, mostly M1 and M6, with only a small proportion eliminated unchanged.


Annual Research Review: Shifting from 'normal science' to neurodiversity in autism science.
Since its initial description, the concept of autism has been firmly rooted within the conventional medical paradigm of child psychiatry. Increasingly, there have been calls from the autistic community and, more recently, nonautistic researchers, to rethink the way in which autism science is framed and conducted. Neurodiversity, where autism is seen as one form of variation within a diversity of minds, has been proposed as a potential alternative paradigm. In this review, we concentrate on three major challenges to the conventional medical paradigm - an overfocus on deficits, an emphasis on the individual as opposed to their broader context and a narrowness of perspective - each of which necessarily constrains what we can know about autism and how we are able to know it. We then outline the ways in which fundamental elements of the neurodiversity paradigm can potentially help researchers respond to the medical model's limitations. We conclude by considering the implications of a shift towards the neurodiversity paradigm for autism science.


The relationship between social support and personal recovery in bipolar disorder.
Mood symptoms may impact on personal recovery and psychosocial functioning in bipolar disorder (BD). This study aimed to explore whether recent mood symptoms were associated with recovery and social support from friends, family, and partners. A cross-sectional survey was conducted with 312 participants who self-reported a confirmed diagnosis of BD and were recruited through social media. A series of one-way analyses of variance (ANOVA) were conducted. Those who reported depression (52%) or hypo/mania (47%) reported lower recovery scores. Those who reported seeing family and >1 friend reported higher recovery scores along with those who were employed, did not receive benefits, and had higher education. Recent hypo/mania symptoms were associated with less family contact. Friends and family may be conducive to recovery, however recent symptoms may have an impact. More research is required to understand how hypo/mania symptoms may influence accessing family support. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Upstream anticoagulation in patients with ST-segment elevation myocardial infarction: a systematic review and meta-analysis.
Parenteral anticoagulation is recommended for all patients presenting with ST-segment elevation myocardial infarction (STEMI) during primary percutaneous coronary intervention (PPCI). Whether upstream anticoagulation improves clinical outcomes is not well established. We conducted a systematic review and meta-analysis of contemporary evidence on parenteral anticoagulation timing for STEMI patients. We performed a systematic search of electronic databases (PubMed, CENTRAL, and Scopus) until December 2022. Studies were eligible if they (a) compared upstream anticoagulation with administration at the catheterization laboratory and (b) enrolled patients with STEMI undergoing PPCI. Efficacy outcomes included in-hospital or 30-day mortality, in-hospital cardiogenic shock (CS), and TIMI flow grade pre- and post-PPCI. Safety outcome was defined as in-hospital or 30-day major bleeding. Overall, seven studies were included (all observational), with a total of 69,403 patients. Upstream anticoagulation was associated with a significant reduction in the incidence of in-hospital or 30-day all-cause mortality (OR 0.61; 95% CI 0.45-0.81; p < 0.001) and in-hospital CS (OR 0.68; 95% CI 0.58-0.81; p < 0.001) and with an increase in spontaneous reperfusion (pre-PPCI TIMI > 0: OR 1.46; 95% CI 1.35-1.57; p < 0.001). Pretreatment was not associated with an increase in major bleeding (OR 1.02; 95% CI 0.70-1.48; p = 0.930). Upstream anticoagulation was associated with a significantly lower risk of 30-day all-cause mortality, incidence of in-hospital CS, and improved reperfusion of the infarct-related artery (IRA). These findings were not accompanied by an increased risk of major bleeding, suggesting an overall clinical benefit of early anticoagulation in STEMI. These results require confirmation in a dedicated randomized clinical trial.


Galactose supplementation enhance sialylation of recombinant Fc-fusion protein in CHO cell: an insight into the role of galactosylation in sialylation.
Sialic acid levels of therapeutic glycoprotein play an important role in plasma half-life. An undesirable decrease of sialic acid content was observed when we increased Fc-fusion protein productivity fourfold in a GS-CHO cell line by bioprocess optimization. We investigated the potential mechanism for the sialic acid content reduction. We found that limited nucleotide sugar precursor and the extracellular sialidase were not responsible for the reduction of the sialic acid content after titer improvement. Oligosaccharide analysis revealed that the lack of protein galactosylation was the potential cause for the reduction of sialic acid content. Thus we validated this notion by evaluated galactose supplementation in 2 L bioreactors. Cell culture performance was not impacted by addition of up to 40 mM galactose except for the glucose consumption rate. Addition of 20 mM galactose to the bioreactor resulted in the increase of 44 % for total sialic acid content and 20.3 % for sialylated glycans. These data were further validated when the process was run on 200 L scaled bioreactor. These data together show that the galactosylation plays an apparent role in sialylation in our current system.


Noninvasive evaluation of intracranial pressure in patients with traumatic brain injury by transcranial Doppler ultrasound.
The purpose of this study was to investigate the relationship between pulsatility index (PI) or optic nerve sheath diameter (ONSD) and intracranial pressure (ICP) in patients with traumatic brain injury (TBI), and the ability of ONSD and ICP to predict intracranial hypertension. A total of 68 patients with TBI were included in this retrospective study. After receiving surgery treatment, they underwent transcranial Doppler ultrasound (TCD). The statistical correlation between PI or ONSD and ICP 1 week after surgery was analyzed. Furthermore, the areas under the curve (AUCs) of ONSD or PI or a combination of them were calculated to predict intracranial hypertension. There was a correlation between ONSD and ICP. This correlation still remained at ONSD ≥ 5 mm. Furthermore, there was a strong correlation between PI and ICP. There was a moderate correlation between ICP and PI on days 3, 4, and 5 after surgery (r = 0.508, p < .001), and a strong correlation on days 6 and 7 after surgery (r = 0.645, p < .001). Moreover, for predicting intracranial hypertension with PI ≥ 1.2 mm or ONSD ≥ 5 mm or a combination of them, the AUC was 0.729, 0.900, and 0.943, respectively (p < .001). The correlation between ONSD or PI and invasive ICP was different with different levels of ICP in different periods in patients with TBI after surgery. When ONSD ≥ 5 mm and PI ≥ 1.2, it could predict elevated ICP more accurately.