Autoregulation of 5-fluorouracil metabolism.
5-Fluorouracil (5-FU) is a commonly used anticancer agent for the treatment of gastrointestinal, head and neck, and breast tumours. This study determined the influence of 5-FU on dihydropyrimidine dehydrogenase (DPD) activity, the enzyme responsible for its in vivo degradation. DPD activity was measured in mononuclear cells obtained prior to and after the administration of 5-FU in 20 patients with colorectal cancer. Following the results from the human studies, DPD activity was measured in Sprague-Dawley rat liver up to 72 h after administration of 5-FU 200 mg/kg as a single injection. Total liver P450 content and the production of testosterone metabolites (indicative of CYP3A activity) were also analysed to determine the specificity of 5-FU-associated alteration in rat liver metabolism. Human mononuclear cell DPD activity decreased by a median of 38.7% following the administration of 5-FU (P = 0.001). 5-FU-induced alterations in rat liver DPD were also observed, with the lowest activity occurring 48 h after injection (50% of control activity; P = 0.009). Rat liver DPD activity returned to near control values by 72 h postinjection. Rat liver total P450 content and CYP3A activity were not significantly different in 5-FU treated or control tissues. Thus, 5-FU demonstrates autoregulation of its metabolism through inhibition of DPD activity. Although this inhibition appears to be specific for DPD, the mechanism for enzyme inhibition is not clear. These findings may aid in the design of 5-FU treatment regimens and provide the basis for further studies into the regulation of DPD.


The Expression of E2F1, p53, and Caspase 3 in the Rat Dorsal Root Ganglia After Sciatic Nerve Transection.
Neurotrauma is among the main causes of human disability and mortality. Nerve injury impairs not only neurons but also causes death of satellite glial cells remote from the injury site. We studied the dynamics of expression of different proapoptotic proteins (E2F1, p53, caspase 3) in the dorsal root ganglia (DRG) of a rat after sciatic nerve transection. TUNEL staining and immunoblotting were used for analysis of cell apoptosis and axotomy-induced biochemical changes. Apoptosis of glial cells was observed at 24 h after sciatic nerve transection and increased on day 7, when apoptosis of some neurons only started. The earliest proapoptotic event in the injured DRG was overexpression of transcription factor E2F1 at 4 h after sciatic nerve transection. This preceded the induction of p53 and cleavage of caspase 3 at 24-h post-axotomy. The nerve injury marker amyloid precursor protein and the nerve regeneration marker GAP-43 were overexpressed in DRG on day 7 after sciatic nerve transection. We also developed a novel fluorescence method for differential visualization of the rat DRG and nerves by means of double staining with propidium iodide and Hoechst 33342 that impart red and blue-green fluorescence, respectively. The present experiments showed that glial cells remote from the nerve transection site were more vulnerable to axotomy than DRG neurons. E2F1 and p53 may be considered promising molecular targets for development of potential neuroprotective agents.


Reexamination of carbodiimide as a possible affinity label for the acetylcholine receptor at the frog neuromuscular junction.
The effects of water-soluble carbodiimide were examined at the frog neuromuscular junction. Acetylcholine sensitivity was measured using a fluid electrode technique and intracellular recording of miniature end-plate potentials. The carbodiimide blocked synaptic sensitivity by a reversible, curare-like action. Irreversible blockade was also observed, probably due to covalent binding. The conditions of reaction and irreversibility suggest that several different residues may be attacked. The inability of cholinergic antagonists to protect the receptor from attack indicates that nonspecific sites, and not the acetylcholine binding site, are involved.


Characterization of the solution reactivity of a basic heme peroxidase from Cucumis sativus.
A basic heme peroxidase has been isolated from cucumber (Cucumis sativus) peelings and characterized through electronic and (1)H NMR spectra from pH 3 to 11. The protein, as isolated, contains a high-spin ferriheme which in the low pH region is sensitive to two acid-base equilibria with apparent pK(a) values of approximately 5 and 3.6, assigned to the distal histidine and to a heme propionate, respectively. At high pH, a new low-spin species develops with an apparent pK(a) of 11, likely due to the binding of an hydroxide ion to the sixth (axial) coordination position of the Fe(III). A number of acid-base equilibria involving heme propionates and residues in the distal cavity also affect the binding of inorganic anions such as cyanide, azide, and fluoride to the ferriheme, as well as the catalytic activity. The reduction potentials of the native protein and of its cyanide derivative, determined through UV-Vis spectroelectrochemistry, result to be -0.320+/-0.015 and -0.412+/-0.010V, respectively. Overall, the reactivity of this protein parallels those of other plant peroxidases, especially horseradish peroxidase. However, some differences exist in the acid-base equilibria affecting its reactivity and in the reduction potential, likely as a result of small structural differences in the heme distal and proximal cavities.


A mathematical consideration for the optimal shell change of hermit crab.
Shell of the adult hermit crab has some important roles for its fitness. In the same time, the shell size often limits the body growth of its owner. To grow the body size larger, the individual must change the shell to another larger shell. If the individual cannot get another larger one, the individual has to suppress the body size growth as the occupied shell size allows. Growth suppression would result in the lower fitness. With a simple mathematical model, we consider the criterion about whether the individual should try to change the shell or not in order to get the higher fitness. We show that the optimality of a shell change behavior has a relation with the body size and the season length for the shell change. They also affect the optimal timing for the shell change. It is implied that the probability of the success in a shell change and the cost for the shell change behavior do not affect the optimal timing for the shell change at all but significantly do the optimality of the behavioral choice.


Dimethyl adipimidate/Thin film Sample processing (DTS); A simple, low-cost, and versatile nucleic acid extraction assay for downstream analysis.
Sample processing, especially that involving nucleic acid extraction, is a prerequisite step for the isolation of high quantities of relatively pure DNA for downstream analyses in many life science and biomedical engineering studies. However, existing methods still have major problems, including labor-intensive time-consuming methods and high costs, as well as requirements for a centrifuge and the complex fabrication of filters and membranes. Here, we first report a versatile Dimethyl adipimidate/Thin film based Sample processing (DTS) procedure without the limitations of existing methods. This procedure is useful for the extraction of DNA from a variety of sources, including 6 eukaryotic cells, 6 bacteria cells, and 2 body fluids in a single step. Specifically, the DTS procedure does not require a centrifuge and has improved time efficiency (30 min), affordability, and sensitivity in downstream analysis. We validated the DTS procedure for the extraction of DNA from human body fluids, as well as confirmed that the quality and quantity of the extracted DNA were sufficient to allow robust detection of genetic and epigenetic biomarkers in downstream analysis.


Apoptotic and anti-angiogenic effects of Salvia triloba extract in prostate cancer cell lines.
Plants, due to their remarkable composition, are considered as natural resources of bioactive compounds with specific biological activities. Salvia genus (Lamiaceae) has been used around the world in complementary medicine since ancient times. We investigated the cytotoxic, apoptotic and anti-angiogenic effects of methanolic Salvia triloba extract (STE) in prostate cancer cells. Cell viability was evaluated by XTT; apoptosis was investigated by DNA fragmentation and caspase 3/7 activity assays. Changes in the angiogenic cytokine levels were investigated by human angiogenesis antibody array. Scratch assay was used to determine the cell motility. STE induced cytotoxicity and apoptosis in a concentration-dependent manner in both cancer cells; however, it was not cytotoxic to normal cells. Cell motility was reduced in PC-3, DU-145 and HUVEC cells by STE treatment. ANG, ENA-78, bFGF, EGF, IGF-1 and VEGF-D levels were significantly decreased by -2.9, -3.7, -1.7, -1.7, -2.0 and -1.8 fold in STE-treated DU-145 cells, however, ANG, IL-8, LEP, RANTES, TIMP-1, TIMP-2 and VEGF levels were significantly decreased by -5.1, -2.0, -2.4, -3.1, -1.5, -2.0 and -2.5 fold in PC-3 cells. These data suggest that STE might be a promising candidate for anti-tumor and anti-angiogenic treatment of prostate cancer.


Phosphorylated HuR shuttles in cycles.
HuR is a ubiquitous RNA-binding protein (RBP) that associates with many mRNAs encoding proliferative proteins. Although predominantly nuclear, HuR translocation to the cytoplasm is linked to its ability to stabilize target mRNAs and modulate their translation. We recently reported that HuR phosphorylation by Cdk1 at S202 (within the HuR hinge region that is necessary for nucleocytoplasmic shuttle) increases HuR association with 14-3-3 and contributes to its nuclear retention. In the next issue of Cell Cycle we report that residue S242 also regulates HuR's cytoplasmic localization, influences cyclin expression, and modulates cell proliferation. Together with evidence of other post-translational HuR modifications, we propose that HuR phosphorylation ensures the timely mobilization of HuR across the nuclear envelope. In this manner, HuR helps to schedule gene expression programs in a cell cycle-dependent manner.


Metabolism of indium chelates attached to monoclonal antibody: minimal transchelation of indium from benzyl-EDTA chelate in vivo.
The metabolism of radiolabeled antibodies is important for radioimmunoimaging and therapy. The loss of indium-111 (111In) from the chelate can pose problems in imaging and increases the radiation dose to normal tissues. We have evaluated the loss in vivo of 111In from Lym-1-benzyl-EDTA-111In (an antibody conjugated with isothiocyanato-benzyl-EDTA) in normal mice. A monoclonal antibody (CHA 255) that binds to benzyl-EDTA-indium chelates, but not to other forms of indium, was used to measure the percent of 111In remaining in the chelate. Four days after injection, 97.4 +/- 2.2% of the 111In in the liver was still in the benzyl-EDTA chelate, as was 99.4 +/- 0.7% of the 111In in the urine, and 99.1 +/- 0.7% of the 111In in the blood. Studies in vitro indicate that a benzyl-EDTA-111In-antibody-chelate conjugate is more stable in human serum than a benzyl-DTPA-111In conjugate, and that both benzyl-chelate conjugates are much more stable than an unsubstituted DTPA conjugate.


Reduction of carbon dioxide on jet spray formed titanium dioxide surfaces.
The photocatalytic reduction of carbon dioxide (CO(2)) on jet spray formed titanium dioxide (TiO(2)) was studied using light-emitting diode (LED) illumination centred at a wavelength of 388 nm. In addition, the photocatalytic reduction of CO(2) under soft X-ray irradiation was also studied. Specifically, the experiments examined the reduction of CO(2) in a gaseous and liquid-gas system using residual gas analysis mass spectrometry. A photochemical reduction of CO(2) was observed over a course of 250 min, with transformation to a major product, C(2)H(3)O(-) (ethenolate), until equilibrium was reached. The product was observed to be surface stabilised, with it reverting back to CO(2) over the course of 100 min without illumination. A proposed free radical mechanism is presented for the formation of this product. A similar effect to that of UV illumination is also observed to occur under the influence of soft X-rays, which presents a potentially significant alternative method for the activation of TiO(2).


Comparison of metabolic abnormalities 48 weeks after switching from highly active antiretroviral therapy containing a non-nucleoside reverse transcriptase inhibitor to Trizivir versus continued highly active antiretroviral therapy.
Forty patients participating in the TRIZAL study were treated with a non-nucleoside reverse transcriptase inhibitor (NNRTI)-containing regimen before being randomly assigned either to continue their baseline therapy or to switch to a triple nucleoside regimen. No difference was observed in treatment efficacy between the two groups, and total cholesterol was observed to improve significantly in the switch group. Switch maintenance may be an appropriate strategy in patients treated with an NNRTI.


Effects of valproate, lamotrigine, and levetiracetam monotherapy on bone health in newly diagnosed adult patients with epilepsy.
The aim of this study was to evaluate the effects of valproate (VPA), lamotrigine (LTG), and levetiracetam (LEV) on bone turnover and bone mineral density (BMD) in newly diagnosed adult patients with epilepsy. Eligible adult patients who were newly diagnosed with epilepsy were treated with VPA, LTG, and LEV. The chemical indicators of bone metabolism and BMD were measured before treatment and 2 years after treatment with different antiseizure medication (ASM) monotherapies. Then, the differences in these parameters before and after treatment were analyzed. One hundred twenty-four patients completed the 2 years follow-up; 43 received monotherapy with VPA, 32 received LTG, and 49 received LEV. Serum parathyroid hormone (PTH), bone alkaline phosphatase (B-ALP), and β-cross-linked C-telopeptide of type I collagen (β-CTX) levels were elevated in adult patients after 2 years of VPA administration; the serum procollagen I intact N-terminal peptide (PINP) level was noticeably higher in patients after LEV treatment than before treatment. Meanwhile, the BMD of the lumbar spine and femoral neck did not change in patients treated with VPA, LTG, and LEV. Valproate altered bone turnover in adult patients with epilepsy, while LTG and LEV did not exert harmful effects on bone health in adult patients.


Activated persulfate for organic chemical degradation: A review.
Activated persulfate reactions have widespread application for groundwater and environmental remediation, as many of these reactions involve destruction of environmental contaminants. Within the last five years, knowledge of activated persulfate degradation reactions has grown to include novel means of activating persulfate for enhanced removal of organic species. These current studies cover a long list of organic analytes, including pharmaceuticals, pesticides, halogenated compounds and dyes. An extensive review of recently published experimental parameters and results for the destruction of organic compounds via activated persulfate is presented. Focus is placed on emerging methodologies and manipulation of traditional activation techniques. Knowledge gaps are identified and discussed, as despite the number of publications on this subject, more broad-reaching guidelines are needed for optimizing applications of activated persulfate in water treatment.


Translocation of the intrauterine contraceptive device: study of thirty-one cases.
Thirty-one cases of a translocated intrauterine contraceptive device were studied. Etiology, diagnosis, management, and prevention are discussed.


Dynamics of inter- and intrastrand hole transport in DNA hairpins.
The dynamics of photoinduced electron transfer has been investigated in DNA hairpins possessing a stilbenedicarboxamide (Sa) electron acceptor, a guanine (G) primary donor, and two adjacent guanines (GG) as secondary donors. Hole transport from G to GG across a single A is more rapid than across AA or T by factors of 20 +/- 7 and 40 +/- 15, respectively. Intrastrand hole transport across a single A is more rapid than interstrand transport by a factor of 7 +/- 3.


Physiological expression of neural marker proteins in the heart of young rats.
The alpha-subunit of a GTP-binding protein Go (Go alpha), is a new marker protein of neurons and neuroendocrine cells. In our previous report, this protein was shown to be also distributed in the cardiac muscle of young rats, although the cardiac muscle is traditionally thought to be a non-neuronal tissue. To evaluate this unusual finding, expression of other neural marker proteins in the heart was examined. Immunohistochemical studies using antibodies against gamma-enolase, neurofilament and Go alpha protein revealed localization of these neural markers only in neurons and neuroendocrine cells in adult rats. In young rats, however, these neural markers were localized not only in these cells but also in the cardiac muscle. An enzyme immunoassay study of enolase isozymes showed that the heart of young rats was rich in neuron-specific gamma-enolase rather than in muscle-specific beta-enolase, while the heart of adult rats was deprived of the former and rich in the latter. These results suggest that the heart of young rats should be understood as a neuroendocrine tissue as well as a muscular tissue.


Increased red blood cell alloimmunization rates in transfused aplastic anemia and myelofibrosis patients.
Red blood cell (RBC) alloimmunization (AI) is a well-known complication of RBC transfusions, which results in the formation of alloantibodies to non-self antigens on donor RBCs, putting patients at risk of transfusion-related complications. The rate of AI with RBC transfusions in the general hospitalized population is estimated to be 2%-3%. However, some patients who are deemed "transfusion-dependent" require regular transfusions of blood products due to persistently low cell counts, putting them at even greater risk of RBC AI and increased morbidity. However, few studies currently exist investigating RBC AI in some transfusion-dependent patient populations, e.g., aplastic anemia (AA) and myelofibrosis (MF). We conducted a 5-year retrospective review to investigate the prevalence of RBC AI, alloantibody incidence, and the number of RBC transfusions in AA and MF patients, who received RBC transfusions within our hospital system. During the study period, 64 AA and 93 MF patients received 1301 and 2766 RBC transfusions, respectively. Compared to the RBC AI rate in the generalized hospitalized patient population (1%-2%), patients with AA and MF had an increased rate of RBC AI incidence rate at 14.1% and 12.9%, respectively. Furthermore, patients with primary MF demonstrated an isolated increased RBC AI incidence rate of 13.3%. The most common alloantibodies produced were anti-E and anti-K. Within our institution, patients with AA and MF had increased incidence rates of RBC AI compared to the general hospitalized patient population and may benefit from an antigen-matched protocol to minimize AI-related complications.


Carbohydrates in xenotransplantation.
The success of allotransplantation has led to an increasing shortage of human organs from deceased donors. This crisis could be resolved by the use of organs from an anatomically suitable animal, such as the pig. The pig and human have, however, been evolving differently for approximately 80 million years, and numerous immunological and physiological barriers have developed that need to be overcome. Differences in carbohydrate epitopes on pig and human cells have been found to play a major role in some of the immunological barriers that have been identified to date. The rejection caused by the presence of galactose-alpha1,3-galactose (Gal) on the pig vascular endothelium and of natural anti-Gal antibodies in humans has recently been prevented by the breeding of pigs that do not express Gal, achieved by knocking out the gene for the enzyme alpha1,3-galactosyltransferase, which was made possible by the introduction of nuclear transfer/embryo transfer techniques. N-glycolylneuraminic acid (the so-called Hanganutziu-Deicher antigen) has been identified as another carbohydrate antigen present in pigs that may need to be deleted if xenotransplantation is to be successful, although some doubt remains regarding its importance. There remain other antipig antibodies against hitherto unidentified antigenic targets that may well be involved in graft destruction; their possible carbohydrate target epitopes are discussed.


Characterization of the Interaction of Daptomycin With Site II on Human Serum Albumin.
Daptomycin, a cyclic lipopeptide antibiotic, is clinically used for the treatment of infections caused by Gram-positive bacteria, including the methicillin-resistant Staphylococcus aureus and the vancomycin-resistant Enterococci. While daptomycin shows high plasma protein binding (90-93%), our knowledge of the binding process is not extensive. To address this issue in more detail, we characterized the binding of daptomycin to plasma proteins and the findings indicate that the association constant for the binding of daptomycin to human serum albumin (HSA) is much higher than that for α1-acid glycoprotein, another plasma protein. Daptomycin was also found to bind to a single site on HSA, which was identified as site II. The findings also suggest that the n-decanoyl moiety of daptomycin penetrates into the hydrophobic pocket of site II and that this acyl moiety interacts with Tyr411 at the entrance to site II. Due to this selective interaction with site II, daptomycin binding was significantly inhibited by drugs (ibuprofen or diazepam) and endogenous compounds (uremic toxins or fatty acids) which also strongly bind to site II. In diseased states, such an inhibition in the binding could result in the pharmacokinetics and therapeutic action of daptomycin being substantially altered.


Long-term outcomes of fractionated stereotactic radiation therapy for pituitary adenomas at the BC Cancer Agency.
To assess the long-term disease control and toxicity outcomes of fractionated stereotactic radiation therapy (FSRT) in patients with pituitary adenomas treated at the BC Cancer Agency. To ensure a minimum of 5 years of clinical follow-up, this study identified a cohort of 76 patients treated consecutively with FSRT between 1998 and 2007 for pituitary adenomas: 71% (54/76) had nonfunctioning and 29% (22/76) had functioning adenomas (15 adrenocorticotrophic hormone-secreting, 5 growth hormone-secreting, and 2 prolactin-secreting). Surgery was used before FSRT in 96% (73/76) of patients. A median isocenter dose of 50.4 Gy was delivered in 28 fractions, with 100% of the planning target volume covered by the 90% isodose. Patients were followed up clinically by endocrinologists, ophthalmologists, and radiation oncologists. Serial magnetic resonance imaging was used to assess tumor response. With a median follow-up time of 6.8 years (range, 0.6 - 13.1 years), the 7-year progression-free survival was 97.1% and disease-specific survival was 100%. Of the 2 patients with tumor progression, both had disease control after salvage surgery. Of the 22 patients with functioning adenomas, 50% (11/22) had complete and 9% (2/22) had partial responses after FSRT. Of the patients with normal pituitary function at baseline, 48% (14/29) experienced 1 or more hormone deficiencies after FSRT. Although 79% (60/76) of optic chiasms were at least partially within the planning target volumes, no patient experienced radiation-induced optic neuropathy. No patient experienced radionecrosis. No secondary malignancy occurred during follow-up. In this study of long-term follow-up of patients treated for pituitary adenomas, FSRT was safe and effective.


Bimanual microincisional cataract surgery technique and clinical outcome.
Bimanual microincisional cataract surgery has been introduced recently as a technique for cataract surgery and it is gaining interest of many cataract surgeons in the world. Over the last few years many changes were made in the phacoemulsification machines and the intraocular lenses design which allowed bimanual microincisional cataract surgery to be safer and more efficient. The purpose of this review is to introduce the technique of bimanual microincisional cataract surgery and to review all the prospective randomized studies comparing bimanual microincisional cataract surgery and standard phacoemulsification in term of safety and efficacy parameters.


Inflammatory pain mediated by a phenotypic switch in brain-derived neurotrophic factor-immunoreactive dorsal root ganglion neurons innervating the lumbar facet joints in rats.
Human low back pain sometimes originates from lumbar facet joints. In human lumbar facet joint inflammation or degeneration, the referred pain is not only expanded into the low back area but also into the leg or foot. The rat L5-L6 facet joint is innervated by the L1-L5 dorsal root ganglia. The presence of brain-derived neurotrophic factor-immunoreactive dorsal root ganglion neurons innervating the L5-L6 facet joint has been confirmed, but changes in the number and distribution of these neurons caused by inflammation have not been studied. Of fluorogold-labeled neurons innervating the L5-L6 facet joint, the proportion of brain-derived neurotrophic factor-immunoreactive dorsal root ganglion neurons was 16% in the control group and 26% in the inflammatory group. The proportion of brain-derived neurotrophic factor-immunoreactive dorsal root ganglion neurons labeled by fluorogold was significantly higher in the inflammatory group than in the control group (P<0.05). The mean cross-sectional area of fluorogold-labeled brain-derived neurotrophic factor-immunoreactive cells increased from 580 to 915 microm(2) in the inflammatory group (P<0.01). Associated with inflammation in facet joints, the increase of brain-derived neurotrophic factor-immunoreactive neurons and the phenotypic switch to large neurons may induce the expansion of facet joint inflammatory pain.


Extension of overall survival beyond objective responses in patients with metastatic renal cell carcinoma treated with high-dose interleukin-2.
In metastatic renal cell carcinoma (mRCC), survival benefit associated with objective response rates of 16-20 % with high-dose interleukin-2 (HDIL-2) is well established and discussed. Based on recently emerged data on efficacy of cancer immunotherapy, we hypothesized that the survival benefit with HDIL-2 extends beyond those achieving objective responses, i.e., to those who achieve stable disease as the best response to treatment. All sequential treatment naïve mRCC patients treated with HDIL-2 at the University of Utah (1988-2013) and University of Michigan (1997-2013) were included. Best responses on treatment were associated with survival outcomes using log-rank and COX regression with a landmark analysis at 2 months. 391 patients (75 % male; median age 55 years) were included and belonged to the following prognostic risk categories: 20 % good, 64 % intermediate, and 15 % poor. Best responses on treatment were complete response (9 %), partial response (10 %), stable disease (32 %), progressive disease (42 %), and not evaluable for response (7 %). No significant differences in progression-free survival (HR 0.74, 95 % CI 0.48-1.1, p = 0.14) or overall survival (HR 0.66, 95 % CI 0.39-1.09, p = 0.11) were observed between patients achieving partial response versus stable disease. Significant differences in progression-free survival (HR 0.13, 95 % CI 0.09-0.22, p < 0.0001) and overall survival (HR 0.33, 95 % CI 0.23-0.48, p < 0.0001) were observed between patients achieving stable disease compared to those with progressive disease and who were not evaluable. Survival benefit with HDIL-2 is achieved in ~50 % patients and extends beyond those achieving objective responses.


Age-Dependent Outcome of First-Line Endovascular and Surgical Revascularization Strategies in Chronic Limb-Threatening Ischemia.
Chronic limb-threatening ischemia (CLTI) has a high mortality and amputation risk even after revascularization. Owing to an aging population the incidence of peripheral arterial disease is rising. However, the current age distribution in patients with CLTI and the impact of age on outcomes remains unclear. For this reason we performed an age-dependent analysis of mortality, morbidity, and amputation-free survival after open surgical revascularization (OSR) and endovascular revascularization therapy (ERT) with identification of risk factors for mortality. Standardized mortality ratios (SMR) were calculated, comparing observed deaths in the study population to expected deaths based on age and gender specific mortality rates of the overall Dutch population. Patients revascularized for the first episode of CLTI between 2013 and 2018 were included in this multicenter retrospective cohort study. The cohort was divided into 2 treatment groups (OSR and ERT) who were each stratified in 3 age groups: early age group < 65 years (G1), middle age group 65-74 years (G2), and elderly age group ≥ 75 years (G3). During the study period 274 limbs (43.9%) were treated with OSR and 350 limbs (56.1%) with ERT. The young population (G1) is only a small part of the whole CLTI population, namely for OSR 22% and ERT 18%. The risk profile of the early age group was characterized by male gender and smoking, whereas the elderly age group was characterized by poor arterial runoff, tissue loss, hypertension, hypercholesterolemia, chronic kidney disease, history of heart disease, chronic obstructive pulmonary disease, and cerebrovascular disease. One year amputation rates were similar between the age groups. However, significantly higher one-year mortality rates were observed in patients ≥ 75 years in comparison to the low mortality rates in patients < 75 years (OSR: G3 19.8% vs. G2 7.1% and G1 6.7%, P = 0.006; ERT: G3 30.7% vs. G2 12.7% and G1 7.8%, P = 0.001). The SMR in this elderly group equaled 3.72 after OSR and 4.04 after ERT. Independent risk factors for mortality after OSR were age, hazard ratio (HR) 1.03 (95% confidence interval [CI] 1.01-1.06; P = 0.006), preoperative hemoglobin level (HR 0.79; 95% CI 0.67-0.92; P = 0.003), tissue loss (HR 1.85; 95% CI 1.22-2.79; P = 0.004), cardiac history (HR 1.56; 95% CI 1.06-2.30; P = 0.024), and development of postoperative delirium (HR 2.75; 95% CI 1.61-4.71; P < 0.001). After ERT we identified age, HR 1.06 (95% CI 1.04-1.08; P < 0.001); preoperative hemoglobin level, HR 0.75 (95% CI 0.65-0.87; P < 0.001); tissue loss, HR 1.71 (95% CI 1.15-2.53; P = 0.008); history of chronic obstructive pulmonary disease, HR 1.99 (95% CI 1.43-1.79; P < 0.001); history of cerebrovascular accident (CVA), HR 1.55 (95% CI 1.09-2.21; P = 0.015); the development of postoperative pneumonia, HR 2.27 (95% CI 1.24-4.16; P = 0.008); postoperative acute kidney injury (AKI), HR 2.42 (95% CI 1.29-4.54; P = 0.006); and postoperative CVA, HR 8.17 (95% CI 1.96-34.15; P = 0.004) as risk factors. The current CLTI population consists mostly of elderly patients and only a small part is younger than 65 years. This shift in the population is important because increasing age is associated with considerable higher one-year mortality rates regardless of the method of revascularization in patients with CLTI. The mortality rates in the elderly group are 3 to 4 times larger than expected in the general population. In relation to the high mortality of the elderly patient, we assume that interventions to prevent postoperative delirium and correct preoperative anemia may be warranted as they appear to be independent risk factors for mortality.


The first cytochrome P450 in ferns. Evidence for its involvement in phytoecdysteroid biosynthesis in Polypodium vulgare.
The fern Polypodium vulgare is a phytoecdysteroid (PE)-producing plant. Cultures of P. vulgare prothalus produce PE, whereas prothalus-derived callus cultures do not. However, this callus line can transform topically applied ecdysone (E) to 20-hydroxyecdysone (20E), which is the last step in the biosynthetic pathway of the main plant PE. This hydroxylation is catalysed by a cytochrome P450 enzyme. E treatment of the callus line results in an increased amount of P450, showing a linear correspondence between the amount of P450 and in vivo E 20-hydroxylation activity, estimated by measuring the bioconversion of E to 20E. This activity can be inhibited by molecules that bind to the P450-heme group. E shows a P450-substrate-binding spectrum with microsomes that overexpress the P450 protein. Finally, a P450 protein was purified from E-treated calli, this being the first P450 to be described in the pterydophyte group.


[Detection of circulating immune complexes in patients with EPH gestosis or intrauterine fetal growth retardation].
Sera from 19 patients with EPH gestosis, 22 patients with hypotrophic newborns, and a control group of ten women with normal pregnancy and fetuses were studied for the presence of circulating immune complexes. Both the C1q solid-phase radioimmunoassay and the polyethylene glycol precipitation test revealed significantly higher levels of circulating immune complexes in both groups of patients.


Reduction of radiation-induced chromosome aberration and apoptosis by dithiothreitol.
We have examined in vitro and in vivo radioprotective effects of a well-known thiol-containing compound, dithiothreitol (DTT). The treatment of both 0.5 and 1 mM of DTT significantly increased clonogenic survival of gamma-ray irradiated Chinese hamster (V79-4) cells. In order to investigate the possible radioprotective mechanism of DTT, we measured gamma-ray induced chromosome aberration by micronucleus assay. In the presence of 0.5 mM or 1 mM DTT, the frequencies of micronuclei were greatly reduced in all dose range examined (1.5-8 Gy). Slightly higher reduction in micronucleus formation was observed in 1 mM DTT-treated cells than in 0.5 mM DTT-treated cells. In addition, incubation with both 0.5 and 1 mM of DTT prior to gamma-ray irradiation reduced nucleosomal DNA fragmentation at about same extent, this result suggests that treatment of DTT at concentrations of 0.5 and 1 mM reduced radiation-induced apoptosis. In vivo experiments, we also observed that DTT treatment reduced the incidence of apoptotic cells in mouse small intestine crypts. In irradiated control group 4.4 +/- 0.5 apoptotic cells per crypt were observed. In DTT-administered and irradiated mice, only 2.1 +/- 0.4 apoptotic cells per crypt was observed. In vitro and in vivo data obtained in this study showed that DTT reduced radiation-induced damages and it seems that the possible radioprotective mechanisms of action of DTT are prevention of chromosome aberration.


Helper-dependent canine adenovirus vector-mediated transgene expression in a neurodegenerative lysosomal storage disorder.
Mucopolysaccharidosis type IIIA (MPS-IIIA) is a severe neurodegenerative lysosomal storage disorder caused by a deficiency of N-sulfoglucosamine sulfohydrolase (SGSH) activity with subsequent accumulation of partially-degraded heparan sulfate and other glycolipids. In this study, we have evaluated a gene therapy approach using a helper-dependent canine adenovirus vector that expresses human SGSH as a means of delivering sustained transgene expression to the brain. Initial testing in a mixed neural cell culture model demonstrated that the vector could significantly increase SGSH activity in transduced cells, resulting in near-normalization of heparan sulfate-derived fragments. While administration of vector by direct injection into the brain of adult MPS-IIIA mice enabled transgene expression for at least 8.5 months post-treatment, it was only in discrete areas of brain. Heparan sulfate storage was reduced in some regions following treatment, however there was no improvement in secondary neuropathological changes. These data demonstrate that helper-dependent canine adenovirus vectors are capable of neural transduction and mediate long-term transgene expression, but increased SGSH expression throughout the brain is likely to be required in order to effectively treat all aspects of the MPS-IIIA phenotype.


Dynamical behavior of epidemiological models with nonlinear incidence rates.
Epidemiological models with nonlinear incidence rates lambda IpSq show a much wider range of dynamical behaviors than do those with bilinear incidence rates lambda IS. These behaviors are determined mainly by p and lambda, and secondarily by q. For such models, there may exist multiple attractive basins in phase space; thus whether or not the disease will eventually die out may depend not only upon the parameters, but also upon the initial conditions. In some cases, periodic solutions may appear by Hopf bifurcation at critical parameter values.


Human papillomavirus (HPV) DNA detection in self-collected urine.
Non-invasive sampling of human genitals to identify high-risk individuals with subclinical oncogenic HPV infection remains a challenge. The study was designed to see if self-collected urine can be used as a simple, non-invasive sampling for screening HPV, particularly for screening/monitoring general population or young adolescents or infants, if they are to be immunized by HPV vaccines. Self-collected urine samples from 100 sexually unexposed college going girls and cervical scrapes from 104 normal healthy sexually active married women were used in this study. Additionally, a group of 55 women were recruited for collecting first urine and later scraped cervical cells to validate urine sampling by directly comparing HPV positivity between the two types of biological specimens. A dry 'paper smear' method for specimen collection and a simple single tube protocol was employed for PCR detection of HPV infection. Out of 100 sexually inexperienced college going girls, only 6 (6%) were positive for HPV infection as revealed by L1 consensus primer and 4 (4%) of them were positive for HPV 16 but none was found positive for HPV 18 DNA. Out of 104 sexually active married women who were cytologically reported as negative by Pap test, 11 (10.5%) were found HPV positive and 7 (6.7%) of them had infection of high-risk HPV type 16. Both urine and later cervical scrapes from a group of 55 women collected as dry 'paper smear' showed perfect matching positivity for HPV between urine and cervical scrape. The use of urine coupled with its dry collection as 'paper smear' facilitating their easy transport, storage and direct PCR detection of HPV DNA opens up an alternative non-invasive approach for population screening of HPV infection, at least in such cases as children and infants in whom invasive samples are difficult to obtain.


Stability, biophysical properties and effect of ultracentrifugation and diafiltration on measles virus and mumps virus.
Measles virus and mumps virus (MeV and MuV) are enveloped RNA viruses used for production of live attenuated vaccines for prophylaxis of measles and mumps disease, respectively. For biotechnological production of and basic research on these viruses, the preparation of highly purified and infectious viruses is a prerequisite, and to meet that aim, knowledge of their stability and biophysical properties is crucial. Our goal was to carry out a detailed investigation of the stability of MeV and MuV under various pH, temperature, shear stress, filtration and storage conditions, as well as to evaluate two commonly used purification techniques, ultracentrifugation and diafiltration, with regard to their efficiency and effect on virus properties. Virus titers were estimated by CCID50 assay, particle size and concentration were measured by Nanoparticle tracking analysis (NTA) measurements, and the host cell protein content was determined by ELISA. The results demonstrated the stability of MuV and MeV at pH <9 and above pH 4 and 5, respectively, and aggregation was observed at pH >9. Storage without stabilizer did not result in structural changes, but the reduction in infectivity after 24 hours was significant at +37 °C. Vortexing of the viruses resulted in significant particle degradation, leading to lower virus titers, whereas pipetting had much less impact on virus viability. Diafiltration resulted in higher recovery of both total and infectious virus particles than ultracentrifugation. These results provide important data for research on all upstream and downstream processes on these two viruses regarding biotechnological production and basic research.


Comparative acute toxicities of surfactants to aquatic invertebrates.
Investigations of the toxicity of surfactants to aquatic invertebrates have been limited primarily to determining the effects on a few species. In this study, the 48-hr LC50 values for three surfactants are reported for six species of aquatic invertebrates. The acute toxicities (LC50) for each surfactant (mg/liter) varied 159 to 580 X and were as follows: C11.8LAS (anionic), 1.7 (Dero sp.) to 270 (Asellus sp.); C14-15 alkylethoxylate (nonionic), 1.0 (Dugesia sp.) to 6.8 (Rhabditis sp.); CTAC (cationic), 0.1 (Gammarus sp.) to 58 (Asellus sp.). When compared to previously developed data, Daphnia magna was typically found to be the most sensitive of all species tested, including fish, to the surfactants.


Sarcomere structures in the rabbit psoas muscle as revealed by fluorescent analogs of phalloidin.
The fluorescent analogs of phalloidin (rhodamine- and fluorescein-phalloidin) bind tightly to the skinned fibres of rabbit psoas muscle at essentially the same sites as phalloidin and mainly stain the known regions of actin localization in the sarcomere: the thin filaments and Z bands. On both sides of the Z bands, unstained zones were observed, suggesting the presence of proteins tightly bound to the thin filaments. In myofibrils which are stretched to such an extent that the actin and myosin filaments do not overlap, stained bands could also be seen at the myosin-band border, which suggests the localization of actin at these sites.


B35 neuroblastoma cells: an easily transfected, cultured cell model of central nervous system neurons.
A panel of neuronal cell lines was derived from tumors of the neonatal rat central nervous system (CNS) in 1974, and two of these lines are in wide use today. Both the B35 and B50 lines offer a number of advantages to researchers who study CNS neurons in culture: they are simple to grow, to differentiate, and to transfect. B50 cells have been used extensively in the study of neuronal cell death, toxicology, and differentiation, whereas B35 cells have proven useful in the molecular analysis of endocytosis and of signaling pathways, in particular those that guide axonal outgrowth and cell motility. This chapter provides protocols for growing and transfecting B35 cells, selecting stable transfectants, exploring protein function using an antisense approach, and assaying cell motility in a Transwell chamber. All of these protocols have been written for researchers who have some skill in basic cell culture techniques, but previous experience with cultured neurons is not required.


Evaluation of several white-rot fungi for the decolorization of a binary mixture of anionic dyes and characterization of the residual biomass as potential organic soil amendment.
Industrial pollution is a great concern for modern society and developing cyclic processes is one of the major challenges. As far as we know, this work is the first to report the use of multiple white-rot fungi species for degrading a binary mixture of anionic dyes under solid state fermentation (SSF) conditions and a further physicochemical characterization of the residual biomass. First, eight white-rot fungi decolorized the dye mixture of brilliant blue FCF and allura red AC adsorbed onto corncob, reaching decolorizations between 11.47% and 87.64%. Then, I. lacteus, B. adusta and T. versicolor, based on the decolorization yield, were selected to evaluate the effect of carbon:nitrogen ratio, moisture content and inoculum quantity on the decolorization percentage. The factorial designs showed that C:N ratio had a negative effect while moisture and inoculum quantity a positive effect. In terms of the kinetics, the three white-rot fungi achieved their maximum decolorization level, around 80.11-86.04%, after 10-12 days. I. lacteus exhibited the highest decolorization percentage, even though only the enzyme manganese peroxidase was detected, with a maximum activity of 6.62 U gds-1 at day 14. Besides, T. versicolor was the only species with laccase activity, with a maximum of 15.94 U gds-1 at day 6 of fermentation. The physicochemical characterization of the biomass allowed to conclude that these aggregates represent a potential organic amendment, as for their significant oxidizable organic carbon (more than 9.5% on wet basis) and essential nutrients content, as well as for their low ash content (less than 1% on wet basis). Finally, for outlining an efficient bioremediation cycle, a cheap and effective methodology for drying the biomass at the end of the SSF process is required.


Running Injuries: The Infrapatellar Fat Pad and Plica Injuries.
When considering knee pain in runners, clinicians differentiate sources of symptoms and determine their cause. Knee problems arise when a runner increases the amount/frequency of the loading through the lower limb. The way the loading is distributed through the knee determines which tissues are abnormally loaded. Knee problems cannot be considered in isolation, requiring a thorough investigation of static and dynamic lower limb mechanics, and footwear and surfaces. This article examines potential sources of knee pain and explores the role of the infrapatellar fat pad and synovial plica in the mechanics of the knee and its involvement in knee symptoms.


Studies on myelin basic protein-specific protein methylase I in various dysmyelinating mutant mice.
Jimpy mice are dysmyelinating mutants characterized by producing near normal levels of myelin basic protein (MBP) in the brain but failing to incorporate these proteins into the myelin sheath. In this study, the activity of MBP-specific protein-arginine N-methyltransferase (protein methylase I) was studied in the brains of normal and jimpy mice of different ages. The enzyme activity varied little with age in normal mice but in 18 and 21 days-old homozygous jimpy mice the activity was reduced by 50% and 75% respectively from the level of their normal littermates. Interestingly, however, heterozygous jimpy mice who are phenotypically normal and quaking mice (a similar dysmyelinating mutant) showed unaltered enzyme levels.


Profiling the antibiotic resistome in soils between pristine and human-affected sites on the Tibetan Plateau.
With increasing pressure from anthropogenic activity in pristine environments, the comprehensive profiling of antibiotic resistance genes (ARGs) is essential to evaluate the potential risks from human-induced antibiotic resistance in these under-studied places. Here, we characterized the microbial resistome in relatively pristine soil samples collected from four distinct habitats on the Tibetan Plateau, using a Smart chip based high-throughput qPCR approach. We compared these to soils from the same habitats that had been subjected to various anthropogenic activities, including residential sewage discharge, animal farming, atmospheric deposition, and tourism activity. Compared to pristine samples, an average of 23.7% more ARGs were detected in the human-affected soils, and the ARGs enriched in these soils mainly encoded resistances to aminoglycoside and beta-lactam. Of the four habitats studied, soils subjected to animal farming showed the highest risks of ARG enrichment and dissemination. As shown, the number of ARGs enriched (a total of 42), their fold changes (17.6 fold on average), and the co-occurrence complexity between ARGs and mobile genetic elements were all the highest in fecal-polluted soils. As well as antibiotics themselves, heavy metals also influenced ARG distributional patterns in Tibetan environments. However, compared to urban areas, the Tibetan Plateau had a low potential for ARG selection and exhibited low carriage of ARGs by mobile genetic elements, even in environments impacted by humans, suggesting that these ARGs have a limited capacity to disseminate. The present study examined the effects of multiple anthropogenic activities on the soil resistomes in relatively pristine environments.


Exploring COVID-19 Vaccine Hesitancy Among Stakeholders in African American and Latinx Communities in the Deep South Through the Lens of the Health Belief Model.
The purpose of this study was to qualitatively explore perceptions related to COVID-19 vaccination intention among African American and Latinx participants and suggest intervention strategies. Ninety minute virtual focus groups (N = 8), segmented by county, race and ethnicity were conducted with stakeholders from 3 vulnerable Alabama counties. Participants (N = 67) were primarily African American and Latinx, at least 19 years, and residents or stakeholders in Jefferson, Mobile, and Dallas counties. Focus groups took place virtually over Zoom. The semi-structured guide explored perceptions of COVID-19, with an emphasis on barriers and facilitators to vaccine uptake. Focus groups lasted approximately 90 minutes and were audio recorded, transcribed, and analyzed by a team of 3 investigators, according to the guidelines of Thematic Analysis using NVivo 12. To provide guidance in the development of interventions to decrease vaccine hesitancy, we examined how themes fit with the constructs of the Health Belief Model. We found that primary themes driving COVID-19 vaccine hesitancy, ordered from most to least discussed, are mistrust, fear, and lack of information. Additionally, interventions to decrease vaccine hesitancy should be multi-modal, community engaged, and provide consistent, comprehensive messages delivered by trusted sources.


Serum Zinc Level and Efficacy of Zinc Therapy in Cutaneous Leishmaniasis: a Systematic Review and Meta-analysis.
Cutaneous leishmaniasis is a parasitic skin disease prevalent in many parts of the world. Zinc has been investigated for its potential role in the immune response against Leishmania parasites. This study aimed to systematically review the literature and conduct meta-analyses to evaluate the serum zinc level and efficacy of zinc therapy in cutaneous leishmaniasis. A comprehensive search of electronic databases was performed to find studies reporting serum zinc levels and the efficacy of zinc therapy in cutaneous leishmaniasis. Meta-analyses were conducted using RevMan software (version 5.4), calculating the mean difference for serum zinc levels and risk ratio for the efficacy of zinc therapy. A total of 11 studies with 1009 participants were evaluated. Five of these studies, comprising 637 participants, examined serum zinc levels; the remaining six, involving 372 individuals, examined the effectiveness of zinc therapy in treating cutaneous leishmaniasis. The results showed that the serum zinc level was significantly lower in cutaneous leishmaniasis patients compared to controls (MD: - 26.65; 95% CI: [- 42.74, - 10.57]; p = 0.001). However, zinc therapy did not demonstrate a significant clinical improvement compared to standard treatment (RR: 0.96; 95% CI: [0.74, 1.23], p = 0.73).


Jejunal Mesenteric Vascularized Lymph Node Transplantation for Lymphedema: Outcomes and Technical Modifications.
The jejunal mesentery supplied by the superior mesenteric vascular tree has emerged as a viable site for vascularized lymph node transplantation. Among other benefits, it has the advantage of avoidance of the risk of donor-site lymphedema. This article reports the technique and outcomes of a novel approach to jejunal mesenteric vascularized lymph node transplantation with flap harvest from the mesenteric root to reduce the risk of small bowel ischemic complications. A consecutive series of patients that underwent jejunal mesenteric vascularized lymph node transplantation to treat upper extremity lymphedema were included. Preoperative and postoperative measurements were taken at fixed intervals using standardized techniques including Perometer volumetry, LDex bioimpedance spectroscopy, the Lymphedema Life Impact Scale, and the Quick Disabilities of the Arm, Shoulder and Hand tool. Demographic, treatment, and outcomes data were collected, and descriptive statistics were used. There were 25 patients included, all of whom had maximized their conservative therapy before undergoing surgery. At 12 months postoperatively reduction in limb volume difference was 36.7 percent (p < 0.001), reduction in LDex score was 41.4 percent (p = 0.0015), and reductions in the Lymphedema Life Impact Scale and Quick Disabilities of the Arm, Shoulder and Hand scores were 55.7 percent (p = 0.0019) and 47.5 percent (p = 0.027), respectively. In 11 patients, there was a history of cellulitis (multiple episodes in eight), and at up to 24 months' follow-up postoperatively there were no episodes reported (p < 0.001). Upper extremity lymphedema can be effectively treated surgically using the jejunal mesenteric vascularized lymph node transplantation, resulting in reduced limb volume and extracellular fluid, and improved patient-reported limb function and outcomes measures compared with optimized conservative therapy alone. Therapeutic, IV.


Knockdown of specific host factors protects against influenza virus-induced cell death.
Cell death is a characteristic consequence of cellular infection by influenza virus. Mounting evidence indicates the critical involvement of host-mediated cellular death pathways in promoting efficient influenza virus replication. Furthermore, it appears that many signaling pathways, such as NF-κB, formerly suspected to solely promote cell survival, can also be manipulated to induce cell death. Current understanding of the cell death pathways involved in influenza virus-mediated cytopathology and in virus replication is limited. This study was designed to identify host genes that are required for influenza-induced cell death. The approach was to perform genome-wide lentiviral-mediated human gene silencing in A549 cells and determine which genes could be silenced to provide resistance to influenza-induced cell death. The assay proved to be highly reproducible with 138 genes being identified in independent screens. The results were independently validated using siRNA to each of these candidates. Graded protection was observed in this screen with the silencing of any of 19 genes, each providing > 85% protection. Three gene products, TNFSF13 (APRIL), TNFSF12-TNFSF13 (TWE-PRIL) and USP47, were selected because of the high levels of protection conferred by their silencing. Protein and mRNA silencing and protection from influenza-induced cell death was confirmed using multiple shRNA clones and siRNA, indicating the specificity of the effects. USP47 knockdown prevented proper viral entry into the host cell, whereas TNFSF12-13/TNFSF13 knockdown blocked a late stage in viral replication. This screening approach offers the means to identify a large number of potential candidates for the analysis of viral-induced cell death. These results may also have much broader applicability in defining regulatory mechanisms involved in cell survival.


Second malignancies after radiotherapy for prostate cancer: systematic review and meta-analysis.
To determine the association between exposure to radiotherapy for the treatment of prostate cancer and subsequent second malignancies (second primary cancers). Systematic review and meta-analysis of observational studies. Medline and Embase up to 6 April 2015 with no restrictions on year or language. Comparative studies assessing the risk of second malignancies in patients exposed or unexposed to radiotherapy in the course of treatment for prostate cancer were selected by two reviewers independently with any disagreement resolved by consensus. Two reviewers independently extracted study characteristics and outcomes. Risk of bias was assessed with the Newcastle-Ottawa scale. Outcomes were synthesized with random effects models and Mantel-Haenszel weighting. Unadjusted odds ratios and multivariable adjusted hazard ratios, when available, were pooled. Second cancers of the bladder, colorectal tract, rectum, lung, and hematologic system. Of 3056 references retrieved, 21 studies were selected for analysis. Most included studies were large multi-institutional reports but had moderate risk of bias. The most common type of radiotherapy was external beam; 13 studies used patients treated with surgery as controls and eight used patients who did not undergo radiotherapy as controls. The length of follow-up among studies varied. There was increased risk of cancers of the bladder (four studies; adjusted hazard ratio 1.67, 95% confidence interval 1.55 to 1.80), colorectum (three studies; 1.79, 1.34 to 2.38), and rectum (three studies; 1.79, 1.34 to 2.38), but not cancers of the hematologic system (one study; 1.64, 0.90 to 2.99) or lung (two studies; 1.45, 0.70 to 3.01), after radiotherapy compared with the risk in those unexposed to radiotherapy. The odds of a second cancer varied depending on type of radiotherapy: treatment with external beam radiotherapy was consistently associated with increased odds while brachytherapy was not. Among the patients who underwent radiotherapy, from individual studies, the highest absolute rates reported for bladder, colorectal, and rectal cancers were 3.8%, 4.2%, and 1.2%, respectively, while the lowest reported rates were 0.1%, 0.3%, and 0.3%. Radiotherapy for prostate cancer was associated with higher risks of developing second malignancies of the bladder, colon, and rectum compared with patients unexposed to radiotherapy, but the reported absolute rates were low. Further studies with longer follow-up are required to confirm these findings.


Selection and evolution of bacteriophages in cellstat.
Objectives of this work were as follows: 1. to establish a laboratory experimental system utilizable in a biophysical approach to molecular evolution; and 2. to provide real world parameters to theories of molecular evolution, especially to Eigen's theory of quasi-species. Secretion type bacteriophage fd of E. coli, closely related phages and artificial chimera phages of fd, and a virulent phage Q beta of E. coli were cultured continuously in a specially designed fermenter called a "cellstat". A phage is cultured in a flow of host bacterial cells. Due to its high dilution rate, the mutant cell could not be selected in the cellstat. It was therefore recognized that the cellstat is suitable for study of the selection and evolution process of a bacteriophage under well-defined environmental conditions without interference from host cell mutations. Population dynamics of bacteriophages of various types in the cellstat were studied theoretically by computer simulation and experimentally. A genetically invariable pure population of phage behaves like an open non-linear chemical reaction system. An invariable mixed population shows a selection process, while a variable population generates an evolution process. Kinetic constants describing the dynamics were determined by curve fitting between the theoretical and the experimental curve obtained from competition experiments and from biological relaxation experiments. One of the most important kinetic parameters thus obtained was the selection coefficient, and its dependence on the base sequence of phage DNA. We drew a local landscape of the selection coefficient near the fd sequence on the base sequence space. From this landscape we were able to confirm the importance of slightly deleterious mutants in molecular evolution. We also confirmed the possibility of developing an evolutionary molecular engineering using a cellstat as an evolution reactor and fd phage as a working replicon. Novelties of this work were as follows: 1. the first stable continuous culture of a bacteriophage was achieved with a cellstat; 2. a local landscape of selection coefficient near the fd sequence on the sequence space was the first experimental drawing of such a map; 3. a biological relaxation method was realized to measure kinetic constants of a biological kinetic process, or molecular evolution; and 4. a practical engineering process of evolutionary molecular engineering was proposed.


Small cell carcinoma of the ovary: clinical and biological study.
Ovarian small cell carcinoma of hypercalcemic type is a rare neoplasm that typically occurs in young females. We describe three cases of ovarian small cell carcinoma of hypercalcemic type occurring in patients aged 24, 37, and 25 years. The first patient had stage IIc disease and had primary surgery with a left salpingo-oophorectomy and omentectomy, followed by chemotherapy with cisplatin and etoposide. Upon a second relapse, pelvic lymphadenectomy was performed, followed by chemotherapy with docetaxel. She is alive for 4 years without any recurrence after the initial treatment. In vitro drug-sensitivity assay revealed the tumor cells were sensitive to taxane and resistant to irinotecan. The second patient had stage III disease and had primary debulking surgery with bilateral salpingo-oophorectomy, simple hysterectomy, omentectomy, low-anterior resection, and lymphadenectomy, followed by chemotherapy with paclitaxel and carboplatin. Four months after the initial treatment she relapsed and died. The tumor cells were sensitive to platinum, taxane, and doxorubicin, but resistant to irinotecan. The third patient had stage III disease and had right salpingo-oophorectomy, omentectomy, and lymphadenectomy, followed by chemotherapy with cisplatin, cyclophosphamide, and doxorubicin. She relapsed 4 months after completing the chemotherapy and died 5 months after secondary debulking surgery. The tumor cells were sensitive to platinum, taxane, and doxorubicin, but resistant to irinotecan. The drug-sensitivity assays suggested that a non-irinotecan, taxane-containing combination might have been suitable as first-line chemotherapy for these patients. However, an effect of such a regimen was seen in only one patient with early-stage disease, and this questions the validity of chemosensitivity testing.


Correction to: Short-Term Symptomatic Relief in Gastroesophageal Reflux Disease: A Comparative Study of Esomeprazole and Vonoprazan.
The original version of the article unfortunately contained percentage errors in second and third paragraphs of GerdQ Score section. Below is the corrected version.


Defining obstructive ventilatory defect in 2015.
There is no clear consensus as to what constitutes an obstructive ventilatory defect (OVD): Is it FEV1/FVC<lower limit of normal (LLN) or<0.70 (respectively, physiological and operational definitions)? To determine, according to the two definitions, the percentage of subjects having an OVD among them explored in a lung function exploration laboratory. This is a retrospective study including 4,730 subjects aged 17-85 years. Subjects were divided according to the presence [physio (+) or operat (+)] or absence [physio (-) or operat (-)] of an OVD, and into younger (<45 years, n=2,076), older (≥45 years, n=2,654), smokers (n=1,208), and non-smokers (n=3,522) groups. For the total sample, the younger and older groups [mean±SD of age (years), respectively, 46.7±14.1; 33.9±7.4, and 56.8±9.1], the 'physiological definition' detected, respectively, 13.46, 43.22, and 5.09% more OVD than the 'operational one' (p<0.05). In addition, the operational definition, compared with the physiological one, overdiagnosed OVD in 2.33 and 0.44% of smokers and non-smokers, respectively, and underdiagnosed it in 4.46% and 29.72% of smokers and non-smokers, respectively (p<0.05). Compared with the group 'physio (-), operat (+)', the 'physio (+), operat (-)' one was younger (74.2±4.7 years vs. 40.9±10.3 years) and had significantly higher FEV1 (62±13% vs. 78±17%) and FVC (71±15% vs. 93±19%). The frequency of OVD much depends on the criteria used for its definition.


Economic Impact of Urological Conditions in Men and Women in Belize.
Urological disease is prevalent in low- and middle-income countries. Concurrently, the inability to maintain employment or provide family care contributes to poverty. We assessed the microeconomic impacts of urological disease in Belize. We conducted a prospective survey-based assessment of patients evaluated during surgical trips by the charity Global Surgical Expedition. Patients completed a survey focusing on impact of urological disease on work and caretaker responsibilities, as well as its economic impact. The primary study outcome was income loss resulting from work impairment or work time missed related to urological disease. Income loss was calculated using the validated Work Productivity and Activity Impairment Questionnaire. A total of 114 patients completed surveys. Overall, 87.7% and 37.2% of respondents reported a negative impact of urological disease on job and caretaking responsibilities, respectively. Nine (7.9%) patients were unemployed secondary to their urological disease. Sixty-one (53.5%) patients provided financial data sufficient for analysis. In this cohort, median weekly income was $250 Belize dollars (approximately $125 United States Dollars), while median weekly cost for urological disease treatment was $25 Belize dollars. Among the 21 (34.5%) patients who missed work due to urological disease, median weekly income loss was $35.6 Belize dollars, representing 55% of their total income. A vast majority (88.6%) of patients reported that cure of urological disease would increase ability to work and/or care for family. In Belize, urological disease results in significant impairment of work and caretaking responsibilities, as well as income loss. Efforts are necessary to provide urological surgeries in low- and middle-income countries as urological disease impacts not only quality of life, but also financial health.


Structural characterization of slow digestion dextrin synthesized by a combination of α-glucosidase and cyclodextrin glucosyltransferase and its prebiotic potential on the gut microbiota in vitro.
Starch-based dietary fibers are at the forefront of functional ingredient research. In this study, a novel water-soluble slow digestion dextrin (SDD) was synthesized by synergy of α-glucosidase and cyclodextrin glucosyltransferase and characterized. Results showed that SDD exhibited high solubility, low viscosity, and resistance to digestive enzymes, and also showed an increased dietary fiber content of 45.7% compared with that of α-glucosidase catalysis alone. Furthermore, SDD was used as the sole carbon source to ferment selected intestinal strains and human fecal microflora in vitro to investigate its prebiotic effects. It was found that SDD could markedly enriched the abundance of Bifidobacterium, Veillonella, Dialister, and Blautia in human gut microflora and yielded higher total organic acid. The combination of α-glucosidase and cyclodextrin glucosyltransferase in this study showed valuable potential for the preparation of a novel slow digestion dextrin with good physicochemical properties and improved prebiotic effects.


Is there a stronger willingness to pay for air quality improvement with high education: new evidence from a survey in China.
As a developing country with the largest population and serious environmental pollution in the world, China has made great efforts in air pollution. Air quality improvement depends not only on government administrative regulations but also on public support, especially how much the public is willing to pay for air quality improvement. Higher education will encourage the public to take action to improve air quality. However, the confirmation of the causality relationship between WTP and education has been missing. This study uses the Chinese General Social Survey (CGSS) to find the relationship between the two, and the conclusions are drawn: OLS regression model and instrumental variable both determine the positive influence of education level on air quality improvement WTP, and Heckman model further verifies the robustness of the conclusion. The positive influence of education level is greater in the groups of men, higher income, higher awareness of acid rain, and more air purifiers, and it has a greater impact on married people in rural areas than in urban areas. The function mechanism of education can improve residents' WTP by increasing regional GDP, promoting urbanization level, expanding afforestation areas, decreasing private car ownership and the number of newly registered civil cars, and reducing sulfur dioxide emissions, nitrogen oxides, and smoke (powder) dust. The total social and economic value of air quality improvement in China is 34.572 billion CNY to 672.42 trillion CNY.


Fractional-order autonomous circuits with order larger than one.
Fractional-order circuit is a kind of circuit which contains fractional-order elements. It has been proved that the fractional-order circuit has some characteristics which are hard to be achieved by integer-order circuits, such as higher degree of freedom in circuit design. For integer-order circuits, there are not only non-autonomous circuits, but also autonomous circuits. Since there are many applications of integral-order autonomous circuits in real world, it is also necessary to explore fractional-order autonomous circuits. However, few research focuses on fractional-order autonomous circuits. Therefore, this paper proposes two kinds of fractional-order autonomous circuits based on fractional-order elements with order larger than one. The corresponding mathematical models are also established based on fractional calculus and their characteristics are analyzed based on circuit theory. Finally, circuit simulation are performed to verify the correctness of theoretical analysis.


Conducting polymer coated single-walled carbon nanotube gas sensors for the detection of volatile organic compounds.
The current work involves fabrication, characterization and subsequent evaluation of poly(3,4-ethylenedioxythiophene) doped with poly(styrene sulfonic acid) (PEDOT:PSS) coated single walled carbon nanotubes (SWNTs) sensors for detecting analytes of interest in industrial manufacturing. By varying the conducting polymer׳s synthesis conditions in terms of charge controlled electropolymerization of the monomer EDOT in presence of the dopant PSS, the sensing performance of the PEDOT:PSS functionalized SWNT sensors was systematically optimized. Electrical characterization in terms of change in resistance, cyclic voltammetry and field-effect transistor measurements was performed to confirm the presence of PEDOT:PSS coating on SWNTs. The optimized sensors exhibited sensing properties over a wide dynamic range of concentrations towards saturated vapors of volatile organic compounds (VOCs) such as methanol, ethanol and methyl ethyl ketone (MEK) at room temperature. The limit of detection of this sensor was found to be 1.3%, 5.95% and 3% for saturated vapors of methanol, ethanol and methyl ethyl ketone (MEK) respectively. In terms of performance, when compared with bare SWNTs, these hybrid sensors exhibited better sensitivity. The underlying mechanism of sensing was also investigated by using them in chemFET mode of sensor configuration.


Structural and mechanistic insights into the CAND1-mediated SCF substrate receptor exchange.
Modular SCF (SKP1-CUL1-Fbox) ubiquitin E3 ligases orchestrate multiple cellular pathways in eukaryotes. Their variable SKP1-Fbox substrate receptor (SR) modules enable regulated substrate recruitment and subsequent proteasomal degradation. CAND proteins are essential for the efficient and timely exchange of SRs. To gain structural understanding of the underlying molecular mechanism, we reconstituted a human CAND1-driven exchange reaction of substrate-bound SCF alongside its co-E3 ligase DCNL1 and visualized it by cryo-EM. We describe high-resolution structural intermediates, including a ternary CAND1-SCF complex, as well as conformational and compositional intermediates representing SR- or CAND1-dissociation. We describe in molecular detail how CAND1-induced conformational changes in CUL1/RBX1 provide an optimized DCNL1-binding site and reveal an unexpected dual role for DCNL1 in CAND1-SCF dynamics. Moreover, a partially dissociated CAND1-SCF conformation accommodates cullin neddylation, leading to CAND1 displacement. Our structural findings, together with functional biochemical assays, help formulate a detailed model for CAND-SCF regulation.


Advanced modeling and optimizing for surface sterilization process of grape vine (Vitis vinifera) root stock 3309C through response surface, artificial neural network, and genetic algorithm techniques.
In vitro, sterilization is one of the key components for proceeding with plant tissue cultures. Since the effectiveness of sterilization has a direct impact on the culture's final outcomes, there is a crucial need for optimization of the sterilization process. However, compared with traditional optimizing methods, the use of computational approaches through artificial intelligence-based process modeling and optimization algorithms provides a precise optimal condition for in vitro culturing. This study aimed to optimise in vitro sterilization of grape rootstock 3309C using RSM, ANN, and genetic algorithm (GA) techniques. In this context, two output responses, namely, Clean Culture and Explant Viability, were optimised using the models developed by RSM and ANN, followed by a GA, to obtain a globally optimal solution. The most influential independent factors, such as HgCl2, NaOCl, AgNO3, and immersion time, were considered input variables. The significance of the developed models was investigated with statistical and non-statistical techniques and was optimised to determine the significance of selected inputs. The optimal clean culture of 91%, and the explant viability of 89% can be obtained from 1.62% NaOCl at a 13.96 min immersion time, according to MLP-NSGAII. Sensitivity analysis revealed that the clean culture and explant viability were less sensitive to AgNO3 and more sensitive to immersion time. Results showed that the differences between the GA predicted and validation data were significant after the performance validation of predicted and optimised sterilising agents with immersion time combinations were tested. In general, GA, a potent methodology, may open the door to the development of new computational methods in plant tissue culture.


Normal fractional flow reserve with a critical stenosis supplying viable myocardium.
A high-grade stenosis to viable myocardium typically produces a positive fractional flow reserve. In the present report, a case of subtotal occlusion of a saphenous vein graft to a viable myocardial territory with normal fractional flow reserve is described, and a possible explanation for this observation is discussed.


[Syndrome X. A paradoxically anonymous identity].
Since the original description, the list of statistically related components, which make up the metabolic syndrome, is in continuous increase. Therefore, different medical groups and their consensus and reports still do not agree on the criteria for its definition and denomination. Although the syndrome is related with high risk of diabetes and cardiovascular disease, the trials have not demonstrated the relationship between hyperinsulinemia and cardiovascular diseases; in consequence it is difficult to conclude whether cardiovascular disease is a part of the syndrome or its consequence. Therefore, it is impossible to accept that the syndrome is synonymous of high risk for cardiovascular disease. These facts may be the reason why its identity hides behind an "X".


Detection of human papillomavirus DNA by AffiProbe HPV-DNA test kit in cervical scrapes or biopsies-histopathologic correlates.
The aim of this study was to evaluate and compare the efficacy of punch biopsies and cervical scrapes in the detection of human papillomavirus (HPV) DNA from the cervix and compare the results with the histopathologic diagnosis. The specimens were collected simultaneously, and HPV DNA was detected using a liquid hybridization test. Biopsies and scrapes were equally efficient, but each detected only two-thirds of all HPV-DNA-positive patients. Thus, the positivity rate increased when both tests were used. Overall, 13% of patients with normal histopathology, 38% of patients with benign atypia, and 66% of patients with squamous intraepithelial lesions (SIL) were HPV-DNA positive. HPV-DNA 16 was found in 54% of HPV-DNA-positive patients with SIL, in 20% of HPV-DNA-positive patients with atypia, and in none of patients with normal histopathology. The liquid hybridization test used in this study detects HPV DNA equally efficiently from both biopsies and scrapes. The test can be performed in 1 working day. However, the sensitivity of the test is low, and it only detects a limited number of HPV types.


Factors associated with acute kidney injury among preterm infants administered vancomycin: a retrospective cohort study.
Vancomycin (VCM) is a widely used antibiotic for the treatment of gram-positive microorganisms, with some nephrotoxic effects. Recent studies have suggested that piperacillin-tazobactam (TZP) aggravates VCM-induced nephrotoxicity in adults and adolescents. However, there is a lack of research investigating these effects in the newborn population. Therefore, this study investigates whether the concomitant use of TZP with VCM use increases the risk of acute kidney injury (AKI) and to explore the factors associated with AKI in preterm infants treated with VCM. This retrospective study included preterm infants with birth weight < 1,500 g in a single tertiary center who were born between 2018 and 2021 and received VCM for a minimum of 3 days. AKI was defined as an increase in serum creatinine (SCr) of at least 0.3 mg/dL and an increase in SCr of at least 1.5 times baseline during and up to 1 week after discontinuation of VCM. The study population was categorized as those with or without concomitant use of TZP. Data on perinatal and postnatal factors associated with AKI were collected and analyzed. Of the 70 infants, 17 died before 7 postnatal days or antecedent AKI and were excluded, while among the remaining participants, 25 received VCM with TZP (VCM + TZP) and 28 VCM without TZP (VCM-TZP). Gestational age (GA) at birth (26.4 ± 2.8 weeks vs. 26.5 ± 2.6 weeks, p = 0.859) and birthweight (750.4 ± 232.2 g vs. 838.1 ± 268.7 g, p = 0.212) were comparable between the two groups. There were no significant differences in the incidence of AKI between groups. Multivariate analysis showed that GA (adjusted OR: 0.58, 95% CI: 0.35-0.98, p = 0.042), patent ductus arteriosus (PDA) (adjusted OR: 5.23, 95% CI: 0.67-41.05, p = 0.115), and necrotizing enterocolitis (NEC) (adjusted OR: 37.65, 95% CI: 3.08-459.96, p = 0.005) were associated with AKI in the study population. In very low birthweight infants, concomitant use of TZP did not increase the risk of AKI during VCM administration. Instead, a lower GA, and NEC were associated with AKI in this population.


Extracellular and immunological actions of zinc.
Zinc is an essential trace element for the immune system, but also very important in other organ systems. Every highly proliferating cell system is dependent on sufficient availability of zinc. During the last decades the influence of zinc on various cell systems have been investigated. Multiple effects of exogenously added zinc have been described in in vitro culture systems and in in vivo systems. However, most of these effects are so far poorly understood, and the dosages used in the in vitro systems are not comparable and sometimes unphysiologically high. Especially in the immune system a number of effects were described and over the last ten years we have come to understand some molecular mechanisms of zinc in this cell system. A zinc deficiency is accompanied by an immunodeficiency, resulting in an increased number of infections. However, the immune function is delicately regulated by zinc, since both increased and decreased zinc levels result in a disturbed immune function. Therefore, zinc supplementation must be accurately supervised. In this review, we discuss the activity of extracellular zinc in four sections. 1. The effect of zinc on different in vitro cell systems, including keratinocytes, osteocytes and leukocytes, and the concentrations of zinc needed for a specific cell response. 2. The modulation of the innate immune system in vitro and in vivo. 3. The role of zinc in the B cell response and antibody production. 4. Effects of zinc on the development and function of T cells.


Anti-AIDS agents. 32. Synthesis and anti-HIV activity of betulin derivatives.
Eleven betulin derivatives were prepared and evaluated for anti-HIV activity in H9 lymphocytes. Compound 4 was found to be the most active with EC50 and TI values of 0.00066 microM and 21,515, respectively.


Effects of urocortin via ion mechanisms or CRF receptors?
Urocortin (UCN), a newly isolated peptide related to hypothalamic corticotrophin releasing factor (CRF) family, had been reported to play biologically diverse roles in several systems such as cardiovascular, reproductive, appetite, stress, and inflammatory responses, etc. It was thought previously to be an endogenous agonist, producing the several actions previously attributed to CRF. But, recently, it was shown to directly reduce L-type calcium currents of acute isolated cardiac myocytes and T-type calcium currents in mouse spermatogenic cells via inhibiting calcium channel instead of binding first to its CRF-R2 receptors. UCN could also reduce the intracellular calcium in vascular smooth muscle cells via inhibiting calcium channel directly. Furthermore, UCN could increase the gene expression of ATP-sensitive potassium channels (K(ATP)) and activate sarcolemmal ATP-sensitive potassium current during normal or hypoxia, which could be inhibited by glibenclamide, a specific K(ATP) blocker. This review will highlight the current novel findings on the ionic mechanisms by which UCN may exert its several actions.


Motivations and barriers towards optimal physical activity level: A community-based assessment of 28 EU countries.
Regular physical activity (PA) is one of the most important determinants of a healthy lifestyle and improved physical and mental well-being. Despite the health benefits of regular PA, the studies show low levels of PA among European adolescents and adults. An increase in physical inactivity has been associated with different personal and environmental factors. This study aimed to investigate positive motivation and barriers to community-based PA. Face-to-face interviews were conducted to collect data on PA, positive motivation and barriers to PA, demographic, and socioeconomic factors. Moderate and vigorous levels of PA were assessed. Regression analysis was applied to estimate the associations for positive and negative reasons for meeting PA recommendations. Data were available for 28,031 adults with a mean age of 48.3 (SD = 18.8) and 51.8% women. Significant variations in individual characteristics were seen between different types of communities. The percentage of participants who reported achieving recommended levels of PA was the lowest in rural areas (44.3%). Regression models showed that most motivational factors had a strong positive association with meeting PA recommendations. We observed the effects of interactions between making acquaintances, having fun and the type of community on meeting PA recommendations. The effects of interactions between the type of community and barriers to PA such as price, risk of injury, disability / illness, and a lack of motivation on PA recommendations were observed. In conclusion, the motivational factors and barriers to PA are associated with the physical environment, and community-based programs and policies for encouraging PA participation are needed.


Synthesis and antimicrobial evaluation of new 1,4-dihydro-4-pyrazolylpyridines and 4-pyrazolylpyridines.
Dialkyl 1,4-dihydro-2,6-dimethylpyridine-3,5-dicarboxylates (1,4-DHP) have now been recognized as vital drugs. Some of these derivatives such as amlodipine, felodipine, isradipine, etc. have been commercialized. In view of wide range of biological properties associated with 1,4-DHP and owing to the biological importance of the oxidation step of 1,4-DHP, we carried out the synthesis and antimicrobial evaluation of new diethyl 1,4-dihydro-2,6-dimethyl-4-(3-aryl-1-phenyl-4-pyrazolyl)pyridine-3,5-dicarboxylates (2a-g) and diethyl 2,6-dimethyl-4-(3-aryl-1-phenyl-4-pyrazolyl)pyridine-3,5-dicarboxylates (3a-g). Synthesis of a series of new diethyl 1,4-dihydro-2,6-dimethyl-4-(3-aryl-1-phenyl-4-pyrazolyl)pyridine-3,5-dicarboxylates (2a-g) has been accomplished by multicomponent cyclocondensation reaction of ethyl acetoacetate, 3-aryl-1-phenyl pyrazole-4-carboxaldehyde (1a-g) and ammonium acetate. The dihydropyridines 2a-g were smoothly converted to new diethyl 2,6-dimethyl-4-(3-aryl-1-phenyl-4-pyrazolyl)pyridine-3,5-dicarboxylates (3a-g) using HTIB ([Hydroxy (tosyloxy)iodo]benzene, Koser's reagent) as the oxidizing agent. The antimicrobial studies of the title compounds, 2a-g &3a-g, are also described.


Brain oxygen utilization measured with oxygen-15 radiotracers and positron emission tomography: generation of metabolic images.
We recently described a PET method for the measurement of local brain tissue oxygen extraction (E) and the local cerebral metabolic rate for oxygen (CMRO2) using 15O-labeled radiotracers. The equation for the calculation of E from measured PET data is mathematically complex and its direct application in the generation of PET images of either E or the CMRO2 on a pixel-by-pixel basis is computationally burdensome. We describe a simplification of this equation which permits the efficient generation of quantitative images.


Combined cyclophosphamide, adriamycin and cis-platinum in advanced ovarian cancer resistant to chlorambucil and cis-platinum.
Seventeen patients with advanced ovarian adenocarcinoma recurring or progressing after prior treatment with chlorambucil and cis-platinum were treated with cyclophosphamide, adriamycin, and cis-platinum (CAP) every 3 weeks. In seven patients there was objective tumor response (41%) with a median response duration of 7.6 months and a survival advantage over nonresponders. Patients whose tumors had previously responded to chlorambucil or cis-platinum were less likely to respond to CAP than those who had not responded. Myelosuppression was the main toxicity, febrile neutropenia occurring after 8% of treatments.


Bradyarrhythmic syncope in a patient with Prinzmetal's variant angina: a case report.
We present the case of a 46-year-old woman with Prinzmetal's angina and syncope due to severe bradyarrhythmias. Dynamic electrical changes were documented on 12-lead 24-hour electrocardiographic (ECG) monitoring. We highlight the importance of continuous ECG monitoring in making the diagnosis and the indication for permanent cardiac pacing in this case.


Agreement between commercial assays for haptoglobin and serum amyloid A in goats.
Haptoglobin (Hp) and serum amyloid A (SAA) are considered as the major acute phase proteins (APPs) in goats. These APPs have been investigated in several studies during the last decade. In most studies, a colorimetric assay for Hp and a solid phase sandwich ELISA for SAA have been used for quantification. In 2015, reference intervals for APPs were determined using a new type of assay, the competitive ELISA (cELISA). Results obtained by the cELISA differed significantly from results obtained by previously used assays. The present study aimed to assess the agreement between so far used assays and cELISAs. Sera of 152 female dairy goats of two Polish national breeds were analysed. The concentration of Hp was determined using a colorimetric assay (Hp-CA) and the cELISA (Hp-cELISA), while a solid phase sandwich ELISA (SAA-sELISA) and the cELISA (SAA-cELISA) were used to measure SAA. Agreement between test results was assessed by preparing Bland-Altman plots, and analyzing 95% limits of agreement (LoA). Finally, the assays for Hp and SAA were compared using 147 and 138 serum samples, respectively, as 5 and 14 paired measurements, respectively, were excluded from agreement analyses to avoid extrapolation of Hp and SAA concentration. Measurements obtained by the Hp-CA and Hp-cELISA showed weak positive correlation (r = 0.24, P = 0.003). Limits of agreement (LoA) ranged from + 1.6 (95% CI 1.4 to 1.8) g/L to - 1.5 (95% CI - 1.7 to - 1.3) g/L. Measurements yielded by the SAA-sELISA and SAA-cELISA did not correlate (r = - 0.01, P = 0.855). LoA ranged from + 14.5 mg/L (95% CI 12.9 to 16.1) to - 8.5 mg/L (95% CI - 10.1 to - 6.9). Agreement between the two types of commercial assays for determination of Hp and SAA concentrations in goats is poor and cELISAs tend to underrate both Hp and SAA concentrations.


Orthographic combinability and phonological consistency effects in reading Chinese phonograms: an event-related potential study.
In this study, event-related potentials (ERPs) were used to trace the temporal dynamics of phonological consistency and phonetic combinability in the reading of Chinese phonograms. The data showed a significant consistency-by-combinability interaction at N170. High phonetic combinability characters elicited greater negativity at N170 than did low phonetic combinability characters, and the combinability effect was only found in the reading of high consistency characters. The results support the phonological mapping hypothesis of the reading-related N170 effect and suggest that the earlier stages of visual word recognition are shaped by the mapping of orthography to phonology even in Chinese. Moreover, our data revealed both consistency and combinability effects at P200 and N400, accounted for by the two-stage framework for visual word recognition. That is, characters with high combinability or high consistency facilitated the earlier stages of orthographic or phonological processing which were due to increased activation at the perceptual level; consequently, less positive P200 was demonstrated. In the later stages, high combinability or high consistency characters were associated with a larger semantic neighborhood, which increased semantic competition and exaggerated the N400 effect. These data support the assumption of radical-based inputs proposed by the lexical constituent model. However, the phonetic consistency effects found at N170 and P200 cannot be reconciled with the current framework of the lexical constituent model. A possible revision will be discussed.


Tracheal tear during laryngopharyngectomy and transhiatal oesophagectomy: a case report.
A case is reported of a tracheal tear developing during laryngopharyngectomy and transhiatal oesophagectomy. Ventilation and oxygenation were managed by removing the tracheostomy tube and advancing a straight cuffed armoured tube via the tracheostomy into one main stem bronchus and applying CPAP to the other bronchus via a Foley catheter. Following gastrointestinal reconstruction, the membranous tracheal tear was repaired via a right lateral thoracotomy.


An emerging role for Serine Protease Inhibitors in T lymphocyte immunity and beyond.
Serine proteases control a wide variety of physiological and pathological processes in multi-cellular organisms, including blood clotting, cancer, cell death, osmo-regulation, tissue re-modeling and immunity to infection. T lymphocytes are required for adaptive cell mediated immunity and serine proteases are not only important for effector function but also homeostatic regulation of cell numbers. Serine Protease Inhibitors (Serpins) are the physiological regulators of serine proteases activity. In this review, I will discuss the role of serpins in controlling the recognition of antigen, effector function and homeostatic control of T lymphocytes through the inhibition of physiological serine protease targets. An emerging view of serpins is that they are important promoters of cellular viability through their inhibition of executioner proteases. This will be discussed in the context of the T lymphocyte survival during effector responses and the development and persistence of long-lived memory T cells. The potent anti-apoptotic properties of serpins can also work against adaptive cell immunity by protecting viruses and tumors from eradication by cytotoxic T cells (CTL). Recent insights from knock-out mouse models demonstrate that these serpins also are required for hematological progenitor cells and so are critical for the development of lineages other than T lymphocytes. Given the emerging role of serpins in multiple aspects of lymphocyte immunity and blood development I will review the progress to date in developing new immunotherapeutic approaches based directly on serpins or knowledge gained from identifying their physiologically relevant protease targets.


Recent insights into the structure of TFIID, its assembly, and its binding to core promoter.
TFIID is a large multiprotein assembly that serves as a general transcription factor for transcription initiation by eukaryotic RNA polymerase II (Pol II). TFIID is involved in the recognition of the core promoter sequences and neighboring chromatin marks, and can interact with gene-specific activators and repressors. In order to obtain a better molecular and mechanistic understanding of the function of TFIID, its structure has been pursued for many years. However, the scarcity of TFIID and its highly flexible nature have made this pursuit very challenging. Recent breakthroughs, largely due to methodological advances in cryo-electron microscopy, have finally described the structure of this complex, both alone and engaged with core promoter DNA, revealing the functional significance of its conformational complexity in the process of core promoter recognition and initiation of Pol II transcription. Here, we review these recent structural insights and discuss their implications for our understanding of eukaryotic transcription initiation.


Chemopreventive and therapeutic modulation of green tea polyphenols on drug metabolizing enzymes in 4-Nitroquinoline 1-oxide induced oral cancer.
Oral cancer is one of the most common cancers in the world. Drugs can modulate the expression of drug metabolizing enzymes and are useful in chemoprevention as well as therapy in cancer. 4-Nitroquinoline 1-oxide (4-NQO) is used to induce oral cancer in the present study. In the present investigation, the effect of green tea polyphenols (GTP) on the activities of cytochrome b5, cytochrome P450, cytochrome b5 reductase (cyt b5 R), cytochrome P450 reductase (cyt P450 R), arryl hydrocarbon hydroxylase (AHH), DT-diaphorase (DTD)(Phase I enzymes) and glutathione-S-transferase (GST) and UDP-glucuronyl transferase (UDP-GT) (Phase II enzymes) were assessed in tongue and oral cavity. In induced rats, there was a decrease in the activity of Phase II enzymes and an increase in the activity of Phase I enzymes. On supplementation of GTP by both simultaneous and post treatment mode (200mg/kg) there was a significant increase in the activity of GST and UDP-GT and a significant decrease in the activity of Phase I enzymes. There was a significant decline in the number of tumors, tumor volume and oral squamous cell carcinoma in both simultaneous and post GTP treated animals relative to 4-NQO induced animals; on comparing simultaneous and post GTP treated animals the number of tumors, tumor volume and oral squamous cell carcinoma was significantly reduced in post treated animals. Thus inhibition of Phase I enzymes could be attributed to the protective efficacy of GTP which deactivates carcinogen and GTP induced the expression of Phase II enzymes that detoxifies the 4-NQO. It can be proposed that GTP plays role as a detoxifying agent by which its modulating role prevented/inhibited the formation of tumor.


Treatment patterns and survival among low-income medicaid patients with head and neck cancer.
Medicaid beneficiaries by definition are low income but they are not necessarily a homogeneous group. No study has assessed differences and disparities among Medicaid beneficiaries with head and neck cancers. To examine predictors of treatment receipt and mortality among Medicaid patients with head and neck cancer. Retrospective cohort study using Medicaid claims linked with cancer registry data for 2 states, California and Georgia, for the years 2002 through 2006. Inpatient and ambulatory care. Medicaid beneficiaries aged 18 to 64 years diagnosed as having head and neck cancer (N = 1308) were included. Descriptive statistics and multivariate regression models analyzed the likelihood of treatment receipt and survival. Receipt of treatment and 12- and 24-month mortality. Fewer than one-third of Medicaid patients with cancer received a diagnosis at an early stage. Overall, black patients were less likely to get surgical treatment and more likely to die than white patients, even after controlling for demographics, stage at diagnosis, and tumor site. Older age and disability status also increased 12-month mortality. Patients in California, who were alive for at least 12 months, have approximately half the odds of dying within 24 months compared with those in Georgia. Concrete steps should be taken to address the significant racial disparities observed in head and neck cancer outcomes among Medicaid beneficiaries. Further research is needed to explore the state-level policies and attributes to examine the startling differences in mortality among the state Medicaid programs analyzed in this study. Pooled comparisons of Medicaid beneficiaries with individuals covered by other types of insurance could mask important disparities among Medicaid beneficiaries, which need to be acknowledged and addressed to improve outcomes for these low-income patients with head and neck cancer.


Aortic angiosarcoma presenting as distal arterial embolization.
Aortic angiosarcoma is a rare, malignant neoplasm of the vasculature, with 24 case reports in the literature. Patients usually present with either aneurysmal or occlusive disease. Treatment consists of en bloc resection of the vessel and contiguous structures with postoperative chemoradiation therapy. Despite surgery and adjunctive measures, prognosis remains dismal as a result of early metastasis and late diagnosis. We report a case of aortic angiosarcoma that presented with distal thromboembolic phenomenon.